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Sample records for suspension cultures monitored

  1. Establishment and characterization of American elm cell suspension cultures

    Science.gov (United States)

    Steven M. Eshita; Joseph C. Kamalay; Vicki M. Gingas; Daniel A. Yaussy

    2000-01-01

    Cell suspension cultures of Dutch elm disease (DED)-tolerant and DED-susceptible American elms clones have been established and characterized as prerequisites for contrasts of cellular responses to pathogen-derived elicitors. Characteristics of cultured elm cell growth were monitored by A700 and media conductivity. Combined cell growth data for all experiments within a...

  2. Callus and cell suspension cultures of carnation

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1972-01-01

    of growth regulators were observed to be 3 × 10−6M indoleacetic acid (JAA) combined with 3 × 10−6M benzylaminopurin (BAP) or 10−6M 2,4-dichlorophenoxy acetic acid (2,4-D) alone. IAA + BAP caused a 100 fold increase in fresh weight over 4 weeks at 25°C. Addition of casein hydrolysate increased growth further....... Cell suspension cultures worked best in media containing 2,4-D in which they had a doubling time of about 2 days. Filtered suspensions were successfully plated on agar in petri dishes, but division was never observed in single cells. The cultures initiated roots at higher concentrations of IAA or NAA...

  3. Studies on suspension culture of virginia mallow

    Directory of Open Access Journals (Sweden)

    Anna Kasprzyk

    2014-04-01

    Full Text Available Virginia mallow (Sida hermaphrodita (L. Rusby belongs to the Malvaceae family. It is a very important industrial and energetic crop (Kasprzyk et al. 2013. In our studies, we used plant cell suspension cultures due to the fact that it is a useful tool to investigate biochemical, molecular and physiological aspects of many cellular functions (Dong et al. 2010. Virginia mallow seeds, obtained from Prof. Borkowska (University of Life Sciences in Lublin, Poland, were used in this investigation to obtain plants which were grown in sterile conditions in the Department of Plant Anatomy and Cytology, Maria Curie-Skłodowska University in Lublin, Poland. The seeds were surface sterilized and washed three times in sterile, distilled water. After 3 weeks of in vitro culture, young seedlings were used as a source of explants (to callus induction. Two types of explants were used to form callus culture: leaf and petiole. Callus tissues were then aseptically transferred to an Erlenmeyer flask with liquid medium and placed on an orbital shaker moving at 120 rpm. The observations of this suspense culture were conducted under light and confocal LSM microscopes. The authors observed that depending on the type of explants and composition of medium, callus tissue has varied in color and character of growth.

  4. Regeneration of soybean via embryogenic suspension culture

    Directory of Open Access Journals (Sweden)

    Droste Annette

    2001-01-01

    Full Text Available In an attempt to establish an alternative plant regeneration system for soybean [Glycine max (L. Merrill] cultivars used in Brazilian breeding programs, ten genotypes were tested for their embryogenic potential. Cotyledons were removed as explants from immature seeds harvested from field-grown plants. After 45 days on induction medium, the number of responding cotyledons and the number of somatic embryos per immature cotyledon were evaluated. The percentage of explants that produced somatic embryos varied from 1 to 70% among cultivars. The average number of somatic embryos produced per cotyledon pair ranged from 0.01 to 10.3 with a mean of 3.4. Suspension cultures were initiated with three Agrobacterium tumefaciens susceptible cultivars. Suspensions were successfully developed from Bragg and IAS5 cultivars. The packed cell volume, in one-month growth, increased 8.1 fold for Bragg and 3.5 fold for IAS5 and the fresh weight increased 6.6 and 2.8 fold, respectively. The cultivars differed for the analysed parameters. All tissue from each cultivar was transferred to the maturation medium and subsequently to the germination medium. The germination frequency was 45.7 and 54.9% for Bragg and IAS5, respectively. Plants were gradually exposed to ambient humidity over one week and then planted in soil. All plants yielded seeds in the greenhouse.

  5. Growth and Plating of Cell Suspension Cultures of Datura Innoxia

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1974-01-01

    malate) or on NO3−-N alone. Dry weight yield was proportional to the amount of nitrate-N added (47 mg/mg N). Filtered suspension cultures containing single cells (plating cultures) could be grown in agar in petri dishes when NAA or 2,4-D were used as growth substances. Cells grew at densities above 500...

  6. In vitro production of azadirachtin from cell suspension cultures of ...

    Indian Academy of Sciences (India)

    PRAKASH KUMAR G

    to affect the growth and metabolism of cultured cells, and have been studied extensively in different species in .... Specific growth rate of neem cell suspensions in altered nitrate: ammonium ratio. MS, Murashige and Skoog medium; MS medium with .... Stimulated caffeine production has been reported in Coffea arabica cell ...

  7. In vitro plant regeneration from embryogenic cell suspension culture ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-05-02

    May 2, 2008 ... plasts in A. adsurgens (Luo and Jia, 1998a, b). There are only few reports available on plant regeneration systems via somatic embryogenesis (Luo et al., 1999; Hou and. Jia, 2004). Here, we report a protocol for plant regenera- tion from embryogenic cell suspension culture of endemic. A. chrysochlorus.

  8. Establishment of the callus and cell suspension culture of ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-05

    Oct 5, 2009 ... The objective of this work was the optimization of the conditions of callus and cell suspension culture of Elaeagnus angustifolia for the production of condensed tannins. The effects of different conditions on the callus growth and the production of condensed tannins were researched. The leaf tissue part of.

  9. Establishment of sorghum cell suspension culture system for ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-03-18

    Mar 18, 2008 ... Key words: Sorghum, proteomics, callus, cell suspension cultures, total soluble protein, secretome. INTRODUCTION. Sorghum, a cereal crop native to Africa, is drought- tolerant, surviving periods of water deficit (Rosenow et al., 1983). The crop is grown in the semi-arid regions of. Africa and Asia primarily ...

  10. Establishment of the callus and cell suspension culture of ...

    African Journals Online (AJOL)

    The objective of this work was the optimization of the conditions of callus and cell suspension culture of Elaeagnus angustifolia for the production of condensed tannins. The effects of different conditions on the callus growth and the production of condensed tannins were researched. The leaf tissue part of E. angustifolia was ...

  11. Production of recombinant proteins in suspension-cultured plant cells.

    Science.gov (United States)

    Plasson, Carole; Michel, Rémy; Lienard, David; Saint-Jore-Dupas, Claude; Sourrouille, Christophe; de March, Ghislaine Grenier; Gomord, Véronique

    2009-01-01

    Plants have emerged in the past decade as a suitable alternative to the current production systems for recombinant pharmaceutical proteins and, today their potential for low-cost production of high quality, much safer and biologically active mammalian proteins is largely documented. Among various plant expression systems being explored, genetically modified suspension-cultured plant cells offer a promising system for production of biopharmaceuticals. Indeed, when compared to other plant-based production platforms that have been explored, suspension-cultured plant cells have the advantage of being totally devoid of problems associated with the vagaries of weather, pest, soil and gene flow in the environment. Because of short growth cycles, the timescale needed for the production of recombinant proteins in plant cell culture can be counted in days or weeks after transformation compared to months needed for the production in transgenic plants. Moreover, recovery and purification of recombinant proteins from plant biomass is an expensive and technically challenging business that may amount to 80-94% of the final product cost. One additional advantage of plant cell culture is that the recombinant protein fused with a signal sequence can be expressed and secreted into the culture medium, and therefore recovered and purified in the absence of large quantities of contaminating proteins. Consequently, the downstream processing of proteins extracted from plant cell culture medium is less expensive, which may/does balance the higher costs of fermentation. When needed for clinical use, recombinant proteins are easily produced in suspension-cultured plant cells under certified, controllable and sterile conditions that offer improved safety and provide advantages for good manufacturing practices and regulatory compliance. In this chapter, we present basic protocols for rapid generation of transgenic suspension-cultured cells of Nicotiana tabacum, Oriza sativa and Arabidopis

  12. Effects of chilling on protein synthesis in tomato suspension cultures

    International Nuclear Information System (INIS)

    Matadial, B.; Pauls, K.P.

    1989-01-01

    The effect of chilling on cell growth, cell viability, protein content and protein composition in suspension cultures of L. esculentum and L. hirsutum was investigated. Cell growth for both species was arrested at 2 degrees C but when cultures were transferred to 25 degree C cell growth resumed. There was no difference in viability between control and chilled cultures of L. esculentum, however, L. hirsutum control cultures exhibited larger amounts of Fluorescein Diacetate induced fluorescence than chilled cultures. 35 S-methionine incorporation into proteins was 2.5-2 times higher in L. hirsutum than in L. esculentum. Quantitative and qualitative differences, in 35 S-methionine labelled proteins, between chilled and control cultures were observed by SDS-PAGE and fluorography. Protein content in chilled cultures decreased over time but then increased when cultures were transferred to 25 degrees C

  13. Embryo forming cells in carrot suspension cultures

    NARCIS (Netherlands)

    Toonen, M.A.J.

    1997-01-01


    Somatic cells of many plant species can be cultured in vitro and induced to form embryos that are able to develop into mature plants. This process, termed somatic embryogenesis, was originally described in carrot (Daucus carota L.). Somatic embryos develop through the same characteristic

  14. Elicitation of Diacetylenic Compounds in Suspension Cultured Cells of Eggplant

    Science.gov (United States)

    Imoto, Setsuko; Ohta, Yoshimoto

    1988-01-01

    Induction of stress metabolites in the suspension cultured cells of eggplant (Solanum melongena L.) was examined. When autoclaved RNase A or nigeran, both of which are nonspecific phytoalexin elicitors in bean cells, were added to the cell culture of eggplant, greatly enhanced levels of three compounds were observed. One of them was cis-pentadeca-6-ene-1,3-diyne-5,15-diol, a novel diacetylenic compound. This compound has considerable fungitoxic activity. Also identified was falcarindiol, another fungitoxic diacetylenic compound previously reported as one of the phytoalexins in infected tomato fruits and leaves. Elicited compounds preferentially accumulated in the culture medium rather than in the cells and decreased to original levels during prolonged culturing. The elicitation of these compounds was closely correlated with cellular damage in terms of the decrease of growth rate and was inhibited by 10 micromolar cycloheximide. PMID:16665862

  15. Isolation and culture of Celosia cristata L cell suspension protoplasts

    Directory of Open Access Journals (Sweden)

    Retno Mastuti

    2003-06-01

    Full Text Available Developmental competence of Celosia cristata L. cell suspension-derived protoplasts was investigated. The protoplasts were isolatedfrom 3- to 9-d old cultures in enzyme solution containing 2% (w/v Cellulase YC and 0.5% (w/v Macerozyme R-10 which was dissolvedin washing solution (0.4 M mannitol and 10 mM CaCl2 at pH 5.6 for 3 hours. The highest number of viable protoplasts was releasedfrom 5-d old culture of a homogenous cell suspension. Subsequently, three kinds of protoplast culture media were simultaneously examinedwith four kinds of concentration of gelling agent. Culturing the protoplasts on KM8p medium solidified with 1.2% agarose significantlyenhanced plating efficiency as well as microcolony formation. Afterwards, the microcalli actively proliferated into friable watery calluswhen they were subcultured on MS medium supplemented with 0.3 mg/l 2,4-D and 1.0 mg/l kinetin. Although the plant regenerationfrom the protoplasts-derived calli has not yet been obtained, the reproducible developmental step from protoplasts to callus in thisstudy may facilitate the establishment of somatic hybridization using C. cristata as one parent.

  16. Full-scale monitoring of wind and suspension bridge response

    Science.gov (United States)

    Snæbjörnsson, J. T.; Jakobsen, J. B.; Cheynet, E.; Wang, J.

    2017-12-01

    Monitoring of real structures is important for many reasons. For structures susceptible to environmental actions, full-scale observations can provide valuable information about the environmental conditions at the site, as well as the characteristics of the excitation acting on the structure. The recorded data, if properly analyzed, can be used to validate and/or update experiments and models used in the design of new structures, such as the load description and modelling of the structural response. Various aspects of full-scale monitoring are discussed in the paper and the full-scale wind engineering laboratory at the Lysefjord suspension bridge introduced. The natural excitation of the bridge comes from wind and traffic. The surrounding terrain is complex and its effect on the wind flow can only be fully studied on site, in full-scale. The monitoring program and associated data analysis are described. These include various studies of the relevant turbulence characteristics, identification of dynamic properties and estimation of wind- and traffic-induced response parameters. The overall monitoring activity also included a novel application of the remote optical sensing in bridge engineering, which is found to have an important potential to complement traditional “single-point” wind observations by sonic anemometers.

  17. The phosphatidylinositol species of suspension cultured plant cells

    International Nuclear Information System (INIS)

    Heim, S.; Wagner, K.G.

    1987-01-01

    Suspension cultured Nicotiana tabacum and Catharanthus roseus cells were labeled with [ 3 H]inositol, the phospholipid fraction extracted and separated by thin layer chromatography. Three different solvent systems and reference compounds were used to assign the different 3 H-labeled species by autoradiography. The ratio of [ 3 H]inositol incorporation into PI, PIP and PIP 2 was found to be 95:4:1; with some preparations a lyso-PI band was obtained which incorporated about a tenth of the label of the PIP band. With Catharanthus roseus cells a very faint band between PI and lyso-PI was detected which could not be assigned to a reference compound. (orig.)

  18. ( Linum usitatissimum L. cv. Modran cell suspension culture

    Directory of Open Access Journals (Sweden)

    Aleksandra Seta-Koselska

    2018-01-01

    Full Text Available Flax ( Linum usitatissimum L. is an ancient crop that is widely cultivated as a source of oil, fiber, and bioactive compounds. Flax fiber is traditionally used in textile industry, linseed oil is processed for industrial oils, paints, varnishes and bio-petroleum. Flaxseeds are also rich in α-linolenic acid and phytochemicals such as lignans. In addition to the commercial aspects, this species has been used widely and readily in biotechnological, developmental, and plant-pathogen interaction studies. Differences in the levels of endogenous hormones in various cultivars of flax significantly affected the intensity of callogenesis and determined the type and concentration of growth regulators necessary for callus production. The aim of our investigation was to optimize the culture conditions for callus formation and cell proliferation in liquid medium of the Polish cultivar of fiber flax – Modran. In the first step, 4 combinations of phytohormones in the medium were tested to obtain established callus tissue suitable for initiation of suspension culture. Next, we investigated the effect of chosen plant growth regulators on cell divisions, fresh and dry weight, and dispersal of callus cells in liquid medium. Fast growing and friable callus was obtained in a modified MS medium supplemented with 0.5 mg/l BAP and 0.1 mg/l NAA. We determined that for the initiation of cell suspension supplementation with 0.5 mg/l BAP and 0.5 mg/l NAA is optimal. The results obtained indicated that high concentration of cytokinin (BAP in liquid medium limited cell proliferation and decreased biomass formation.

  19. Physiological responses of suspension cultures of Catharanthus roseus to aluminum: changes in polyamines and inorganic ions

    Science.gov (United States)

    Xinhua Zhou; Rakesh Minocha; Subhash C. Minocha

    1995-01-01

    The effects of aluminum (Al) treatment on polyamines were studied using suspension cultures of Madagascar periwinkle [Catharanthus roseus (L.) G. Don]. The addition of A1 (0.2, 0.5, 1.0 mM) to the suspension cultures caused a significant increase in putrescine within 24h only in freshly transferred cells. By contrast, Al treatment reduced putrescine...

  20. Phenylpropanoid Metabolism in Suspension Cultures of Vanilla planifolia Andr. 1

    Science.gov (United States)

    Funk, Christoph; Brodelius, Peter E.

    1990-01-01

    Feeding of cinnamic acid and ferulic acid to non-treated and chitosan-treated cell suspension cultures of Vanilla planifolia resulted in the formation of trace amounts of p-hydroxy benzoic acid (5.2 micrograms per gram fresh weight of cells) and vanillic acid (6.4 micrograms per gram fresh weight of cells), respectively. Addition of a 4-hydroxycinnamate: CoA-ligase inhibitor, 3,4-(methylenedioxy)-cinnamic acid (MDCA), resulted in a reduced biosynthesis of ligneous material with a simultaneous significant increased vanillic acid formation (around 75 micrograms per gram fresh weight of cells). A K1 of 100 micromolar for 4-hydroxycinnamate: CoA-ligase in a crude preparation was estimated for this inhibitor. It is suggested that the conversion of cinnamic acids into benzoic acids does not involve cinnamoyl CoA esters as intermediates. Feeding of 14C-cinnamic acid and 14C-ferulic acid to cells treated with MDCA indicate that cinnamic acid, but not ferulic acid, is a precursor of vanillic acid in these cultivated cells of V. planifolia. PMID:16667725

  1. Phenylpropanoid Metabolism in Suspension Cultures of Vanilla planifolia Andr. 1

    Science.gov (United States)

    Funk, Christoph; Brodelius, Peter E.

    1990-01-01

    Feeding of 4-methoxycinnamic acid, 3,4-dimethoxycinnamic acid and 3,4,5-trimethoxycinnamic acid to cell suspension cultures of Vanilla planifolia resulted in the formation of 4-hydroxybenzoic acid, vanillic acid, and syringic acid, respectively. The homologous 4-methoxybenzoic acids were demethylated to the same products. It is concluded that the side chain degrading enzyme system accepts the 4-methoxylated substrates while the demethylation occurs at the benzoic acid level. The demethylating enzyme is specific for the 4-position. Feeding of [O-14C-methyl]-3,4-dimethoxycinnamic acid revealed that the first step in the conversion is the glycosylation of the cinnamic acid to its glucose ester. A partial purification of a UDP-glucose: trans-cinnamic acid glucosyltransferase is reported. 4-Methoxy substituted cinnamic acids are better substrates for this enzyme than 4-hydroxy substituted cinnamic acid. It is suggested that 4-methoxy substituted cinnamic acids are intermediates in the biosynthetic conversion of cinnamic acids to benzoic acids in cells of V. planifolia. PMID:16667674

  2. Optimizing the transient transfection process of HEK-293 suspension cells for protein production by nucleotide ratio monitoring

    DEFF Research Database (Denmark)

    de Los Milagros Bassani Molinas, Maria; Beer, Christiane; Hesse, F

    2014-01-01

    Large scale, transient gene expression (TGE) is highly dependent of the physiological status of a cell line. Therefore, intracellular nucleotide pools and ratios were used for identifying and monitoring the optimal status of a suspension cell line used for TGE. The transfection efficiency upon...... polyethyleneimine (PEI)-mediated transient gene delivery into HEK-293 cells cultured in suspension was investigated to understand the effect of different culture and transfection conditions as well as the significance of the culture age and the quality of the cell line used. Based on two different bicistronic model...... plasmids expressing the human erythropoietin gene (rHuEPO) in the first position and green fluorescent protein as reporter gene in the second position and vice versa, a completely serum-free transient transfection process was established. The process makes use of a 1:1 mixture of a special calcium...

  3. Structure and organ specificity of an anionic peroxidase from Arabidopsis thaliana cell suspension culture

    DEFF Research Database (Denmark)

    Ostergaard, L; Abelskov, A K; Mattsson, O

    1996-01-01

    The predominant peroxidase (pI 3.5) (E.C. 1.11.1.7) of an Arabidopsis thaliana cell suspension culture was purified and partially sequenced. Oligonucleotides were designed and a specific probe was obtained. A cDNA clone was isolated from an Arabidopsis cell suspension cDNA library and completely...

  4. A population balance equation model of aggregation dynamics in Taxus suspension cell cultures.

    Science.gov (United States)

    Kolewe, Martin E; Roberts, Susan C; Henson, Michael A

    2012-02-01

    The nature of plant cells to grow as multicellular aggregates in suspension culture has profound effects on bioprocess performance. Recent advances in the measurement of plant cell aggregate size allow for routine process monitoring of this property. We have exploited this capability to develop a conceptual model to describe changes in the aggregate size distribution that are observed over the course of a Taxus cell suspension batch culture. We utilized the population balance equation framework to describe plant cell aggregates as a particulate system, accounting for the relevant phenomenological processes underlying aggregation, such as growth and breakage. We compared model predictions to experimental data to select appropriate kernel functions, and found that larger aggregates had a higher breakage rate, biomass was partitioned asymmetrically following a breakage event, and aggregates grew exponentially. Our model was then validated against several datasets with different initial aggregate size distributions and was able to quantitatively predict changes in total biomass and mean aggregate size, as well as actual size distributions. We proposed a breakage mechanism where a fraction of biomass was lost upon each breakage event, and demonstrated that even though smaller aggregates have been shown to produce more paclitaxel, an optimum breakage rate was predicted for maximum paclitaxel accumulation. We believe this is the first model to use a segregated, corpuscular approach to describe changes in the size distribution of plant cell aggregates, and represents an important first step in the design of rational strategies to control aggregation and optimize process performance. Copyright © 2011 Wiley Periodicals, Inc.

  5. A phytochemical study of lignans in whole plants and cell suspension cultures of Anthriscus sylvestris

    NARCIS (Netherlands)

    Koulman, A; Kubbinga, M.E.; Batterman, S; Woerdenbag, H.J.; Pras, N.; Woolley, J.G.; Quax, Wim

    2003-01-01

    In the roots of Anthriscus sylvestris 12 different lignans were detected. Arctigenin, dimethylmatairesinol, dimethylthujaplicatin, podophyllotoxin, 7-hydroxyyatein and 7-hydroxyanhydropodorhizol have not been previously reported to be present in A. sylvestris. In the cell suspension cultures, which

  6. Development of friable embryogenic callus and embryogenic suspension culture systems in cassava (Manihot esculenta Crantz)

    Science.gov (United States)

    Taylor, N J; Edwards, M; Kiernan, R J; Davey, C D; Blakesley, D; Henshaw, G G

    1996-06-01

    Procedures for the production of a new and highly prolific embryogenic culture system have been developed in cassava. The importance of the basal salts and type of auxin in controlling the development of cassava embryogenic tissues has been demonstrated, with culture on Gresshoff and Doy basal medium in the presence of 4-amino-3,5,6,trichloro-picolinic acid (picloram) inducing the formation of friable embryogenic callus from which highly totipotent embryogenic suspension cultures could be established. Plants have been regenerated from these cultures. The availability of embryogenic suspension cultures is considered to have important implications for the application of genetic transformation and other biotechnologies in the agronomic improvement of cassava.

  7. Cell suspension concentration monitoring by using a miniaturized serial high frequency SAWR sensor

    OpenAIRE

    Li, Jian; Feng, Hailin; Fang, Yiming

    2015-01-01

    In this paper, a miniaturized cell suspension concentration monitoring method was investigated. The sensing unit was a carbon screen-printed electrode (CSPE) in serial with a 433MHz vacuum-packaged surface acoustic wave resonator (SAWR). SAWR provided a stable and high operating frequency, which helps to keep the stability and sensitivity of the monitoring system. Living cells suspensions in different concentrations were prepared and dropcast on CSPE. Frequency responses of the sensor were re...

  8. An efficient method for the establishment of cell suspension cultures in potato (Solanum tuberosum L.)

    International Nuclear Information System (INIS)

    Sajid, Z.A.

    2016-01-01

    Cell suspension cultures offers an In vitro system that can be used as a tool for various studies involving mutant selection, mass propagation, protoplast isolation, gene transfer and selection of cell-lines which are resistant to various biotic or abiotic stresses. Research work on the development of cell suspension cultures was carried out to establish the most efficient method in Potato (cv. Desiree). Healthy, well-proliferating tissues from different types of callus cultures (compact, friable, embryogenic or non-embryogenic) were inoculated on various media combinations, i.e., MS, MS2 or AA liquid medium containing 18.09 micro M 2, 4-D. A fixed quantity (0.5-1.0 g) of callus tissue from 60-day-old callus cultures was transferred to 10-25 ml of liquid medium in 100 ml Erlenmeyer flask. Cultures were placed on an orbital shaker and agitated at different speeds (75, 100 or 125 rpm) under 16-h photoperiod at 25 ± 2 degree C. Medium was changed after every 3 days and fractionated tissue was filtered after every 6 days through sterile mesh (100-800 micro m) to develop a cell-line by transferring resulting suspension to fresh medium under the same conditions. Results indicated that eight-week-old translucent, friable, off-white callus cultures were an excellent starting material for the initiation of homogeneous cell suspension cultures as compared to other tested sources. Of the three tested media (MS, MS2 or AA medium containing 18.09 micro M 2, 4-D), MS2 was found to be a better medium for the initiation of cell suspension cultures. Cell suspension cultures, placed in 16-h photoperiod at 25 ± 2 degree C and agitated at 120 rpm using a gyratory shaker showed excellent results. Several other factors influencing quick establishment of cell suspension cultures in this cultivar are also discussed in this communication. (author)

  9. Enhancement effect of shikonin in cell suspension culture and transfermanant culture by radiation application

    International Nuclear Information System (INIS)

    Kim, Jae Sung; Lee, Young Keun; Chung, Byung Yeoup; Lee, Young Bok; Hwang Hye Yeon

    2004-10-01

    The cell lines 679, 679-29 and 622-46 of L. erythrorhizon could be selected on LS agar medium for the production shikonin in cell suspension culture. The shikonin was increased moderately in suspension culture of cell line 622-46 in LS liquid medium containing BA 2 mg·L -1 and IAA 0.2 mg·L -1 in the dark, and was increased by adding 1 μM Cu 2+ and 100 μM methyl jasmonate The accumulation of shikonin in the liquid medium was increased significantly by 2 Gy irradiation to callus of cell line 622-46 and culture in LS liquid medium containing BA 2 mg·L -1 and IAA 0.2 mg·L -1 in the dark and shikonin in cell debris was higher by 16 Gy irradiation. The activity of p-hydroxybenzoate geranyltransferase was increased by irradiation of 2 Gy and 16 Gy of γ radiation. Seedling hypocotyles of L. erythrorhizon were infected with Agrogacterium rhizogenes strain 15834 harboring a binary vector with an intron bearing the GUS (β-glucuronidase) gene driven by cauliflower mosaic virus (CaMV) 35S promotor as well as the HPT (hygromycin phosphotransferase) gene as the selection marker. Hairy roots isolated were hygromycin resistant and had integrated GUS gene in DNA. The root tip grown on M-9 medium showed normal pigment production pattern in border cells and root hairs

  10. Growth and accumulation of flavan-3-ol in Camellia sinensis through callus culture and suspension culture method

    Directory of Open Access Journals (Sweden)

    Sutini Sutini

    2017-02-01

    Full Text Available This study was aimed to assess flavan-3-ol biomass in C. sinensis through callus cultures and suspension cultures derived from leaf explants. Callus initiation of both cultures were using Murashige and Skoog medium were enriched with plant growth regulators Naphtha-lene Acetic Acid 3.0 mg/L and kinetin 2.0 mg/L. The procedures in this study were: (1 callus initiation by cutting the leaves of C. sinen-sis shoots then planted on Murashige and Skoog medium that were enriched with plant growth regulators, (2 sub callus culture on fresh medium that enriched with the same growth regulators, (3 suspension culture initiation of liquid callus, (4 growth examination of callus and suspension cultures in week 12, (5 examination of qualitative-quantitative content of flavan-3-olin suspension cultures at week 4. The results show that suspension cultures contain biomass flavan-3-ol that increase in the same manner of the increase of callus age and weight

  11. In vitro production of azadirachtin from cell suspension cultures of ...

    Indian Academy of Sciences (India)

    The present study aimed to elucidate the effect of nutritional alteration on biomass content and azadirachtin production in cell suspensions of the elite neem variety crida-8. Variations in total nitrogen availability in the medium in terms of different ratios of nitrate:ammonium showed that the ratio 4:1 revealed a profound effect, ...

  12. Establishment of embryogenic suspension cultures of Pinus radiata ...

    African Journals Online (AJOL)

    The development of embryonal suspensor mass (ESM) from immature embryos of Pinus radiata on a solidified growth medium containing 0, 5 mgl -1 benzyladenine, 3, 0 mgl -1 2, 4-dichlorophenoxyacetic acid, 500 mgl -1 casein hydrolysate and 250 mgl -1 L-glutamine was used as inoculum to establish cell suspension ...

  13. Regulation of Cytoplasmic and Vacuolar Volumes by Plant Cells in Suspension Culture

    DEFF Research Database (Denmark)

    Owens, Trevor; Poole, Ronald J

    1979-01-01

    Quantitative microscopical measurements have been made of the proportion of cell volume occupied by cytoplasm in a cell suspension culture derived from cotyledons of bush bean (cv. Contender). On a 7-day culture cycle, the content of cytoplasm varies from 25% at the time of transfer to 45...

  14. Development of a scalable suspension culture for cardiac differentiation from human pluripotent stem cells

    Directory of Open Access Journals (Sweden)

    Vincent C. Chen

    2015-09-01

    Full Text Available To meet the need of a large quantity of hPSC-derived cardiomyocytes (CM for pre-clinical and clinical studies, a robust and scalable differentiation system for CM production is essential. With a human pluripotent stem cells (hPSC aggregate suspension culture system we established previously, we developed a matrix-free, scalable, and GMP-compliant process for directing hPSC differentiation to CM in suspension culture by modulating Wnt pathways with small molecules. By optimizing critical process parameters including: cell aggregate size, small molecule concentrations, induction timing, and agitation rate, we were able to consistently differentiate hPSCs to >90% CM purity with an average yield of 1.5 to 2 × 109 CM/L at scales up to 1 L spinner flasks. CM generated from the suspension culture displayed typical genetic, morphological, and electrophysiological cardiac cell characteristics. This suspension culture system allows seamless transition from hPSC expansion to CM differentiation in a continuous suspension culture. It not only provides a cost and labor effective scalable process for large scale CM production, but also provides a bioreactor prototype for automation of cell manufacturing, which will accelerate the advance of hPSC research towards therapeutic applications.

  15. Production of secondary metabolites trimethyl xanthina by Camellia sinensis L suspension culture

    Science.gov (United States)

    Sutini, Sodiq, Mochamad; Muslihatin, Wirdhatul; Indra, Mochamad Rasjad

    2017-06-01

    Bioactive trimethyl xanthina can be obtained from the plant Camellia sinensis L. To obtain bioactive plant of which there are several hurdles for instance to wait up to five years to be harvested, also it needs land at a certain height from the sea level. Therefore, the production of secondary metabolites trimethyl xanthina need to be developed with suspense culture techniques. The purpose of this study obtained the production of bioactive trimethyl xanthina way culturally suspense in large scale with a relatively short time, potentially as anti-oxidants. Research methods include: (1) initiation of callus from pieces of leaves, shoots the youngest of the plant Camellia sinensis L in the media MS with the optimization of the addition of growth regulators, (2) the subculture of callus on media and plant growth regulator that is equal to the stage of initiation, (3) initiation of suspension culture using explants of callus Camellia sinensis L, (4) Analysis of secondary metabolites trimethyl xanthina growth in suspension culture, (5) the isolation and identification of trimethyl xanthina qualitatively and quantitatively using thin layer chromatography/high performance chromatography column. The results of the study suspension cultures containing bioactive trimethyl xanthina candidates that can be used as an antioxidant.

  16. Cytogenetic studies of Haplopappus gracilis in both callus and suspension cell cultures.

    Science.gov (United States)

    Ashmore, S E; Shapcott, A S

    1989-08-01

    Investigations have been carried out on karyotype change in both callus and suspension cell cultures of Haplopappus gracilis (2n=4). It has been found that polyploidization arises directly in culture to give up to six times the normal diploid chromosome number in some cultures. In polyploid cultures, both chromosome loss and chromosome rearrangements occur to give rise to aneuploid karyotypes displaying chromosomes which differ in morphology from the diploid set. Whole or partial chromosome loss has been observed in the form of lagging chromosomes and chromosome bridges at anaphase, and micronuclei, ring chromosomes and chromosome fragments at other stages in mitosis. C-banded preparations have confirmed the occurrence of chromosomal rearrangements. Comparative investigations suggest that (i) more polyploidy occurs in callus cultures than in suspension cell cultures, and (ii) the presence of cytokinin (kinetin) in the culture medium may reduce the extent of karyotype change.

  17. Differentiation and selection of hepatocyte precursors in suspension spheroid culture of transgenic murine embryonic stem cells.

    Directory of Open Access Journals (Sweden)

    Elke Gabriel

    Full Text Available Embryonic stem cell-derived hepatocyte precursor cells represent a promising model for clinical transplantations to diseased livers, as well as for establishment of in vitro systems for drug metabolism and toxicology investigations. This study aimed to establish an in vitro culture system for scalable generation of hepatic progenitor cells. We used stable transgenic clones of murine embryonic stem cells possessing a reporter/selection vector, in which the enhanced green fluorescent protein- and puromycin N-acetyltransferase-coding genes are driven by a common alpha-fetoprotein gene promoter. This allowed for "live" monitoring and puromycin selection of the desired differentiating cell type possessing the activated alpha-fetoprotein gene. A rotary culture system was established, sequentially yielding initially partially selected hepatocyte lineage-committed cells, and finally, a highly purified cell population maintained as a dynamic suspension spheroid culture, which progressively developed the hepatic gene expression phenotype. The latter was confirmed by quantitative RT-PCR analysis, which showed a progressive up-regulation of hepatic genes during spheroid culture, indicating development of a mixed hepatocyte precursor-/fetal hepatocyte-like cell population. Adherent spheroids gave rise to advanced differentiated hepatocyte-like cells expressing hepatic proteins such as albumin, alpha-1-antitrypsin, cytokeratin 18, E-cadherin, and liver-specific organic anion transporter 1, as demonstrated by fluorescent immunostaining. A fraction of adherent cells was capable of glycogen storage and of reversible up-take of indocyanine green, demonstrating their hepatocyte-like functionality. Moreover, after transplantation of spheroids into the mouse liver, the spheroid-derived cells integrated into recipient. These results demonstrate that large-scale hepatocyte precursor-/hepatocyte-like cultures can be established for use in clinical trials, as well as in

  18. Determination of cardiac glycosides and total phenols in different generations of Securigera securidaca suspension culture

    Directory of Open Access Journals (Sweden)

    Z. Tofighi

    2016-04-01

    Full Text Available Background and objectives: The seeds of Securigera securidaca (L. Deg. & Dorf. (Fabaceae are used as anti-diabetic remedy in Iranian folk medicine. The aim of the present study was to establish the callus and suspension culture of S. securidaca seeds for the first time and to determine the major secondary metabolites including cardiac glycosides and total phenols. Methods: The culture of S. securidaca from seeds was initiated in hormone-supplemented MS medium containing 1 and 0.1 ppm 2, 4-D solution for solid and suspension cultures, respectively, sucrose and vitamins (B1, B2, B6, Folic acid, Biotin, Nicotinamide and Ca pantothenate at 25 °C and 12 h photoperiods. The cardiac glycosides were determined based on the calibration curve of securidaside which was isolated from the seeds extract of S. securidaca. Total phenolic compounds of different generations of suspension culture were determined using Folin Ciocalteu reagent. Results: Callus culture of S. securidaca was grown light cream to pale yellow in color and soft in texture while the cells of suspension culture grew cream to yellow with isolated cells and small aggregates. The production of cardiac glycosides in the 7th generation were more than the seeds extract (p

  19. The effects of cadmium chloride on secondary metabolite production in Vitis vinifera cv. cell suspension cultures.

    Science.gov (United States)

    Cetin, Emine Sema; Babalik, Zehra; Hallac-Turk, Filiz; Gokturk-Baydar, Nilgun

    2014-09-23

    Plant secondary metabolites are possess several biological activities such as anti-mutagenic, anti-carcinogenic, anti-aging, etc. Cell suspension culture is one of the most effective systems to produce secondary metabolites. It is possible to increase the phenolic compounds and tocopherols by using cell suspensions. Studies on tocopherols production by cell suspension cultures are seldom and generally focused on seed oil plants. Although fresh grape, grape seed, pomace and grape seed oil had tocopherols, with our best knowledge, there is no research on tocopherol accumulation in the grape cell suspension cultures. In this study, it was aimed to determine the effects of cadmium chloride treatments on secondary metabolite production in cell suspension cultures of grapevine. Cell suspensions initiated from callus belonging to petiole tissue was used as a plant material. Cadmium chloride was applied to cell suspension cultures in different concentration (1.0 mM and 1.5 mM) to enhance secondary metabolite (total phenolics, total flavanols, total flavonols, trans-resveratrol, and α-, β-, γ- δ-tocopherols) production. Cells were harvested at two days intervals until the 6th day of cultures. Amounts of total phenolics, total flavanols and total flavonols; trans-resveratrol and tocopherols (α-, β-, γ- and δ-tocopherols) and dry cell weights were determined in the harvested cells. Phenolic contents were significantly affected by the sampling time and cadmium concentrations. The highest values of total phenolic (168.82 mg/100 g), total flavanol (15.94 mg/100 g), total flavonol (14.73 mg/100 g) and trans-resveratrol (490.76 μg/100 g) were found in cells treated with 1.0 mM CdCl2 and harvested at day 2. Contents of tocopherols in the cells cultured in the presence of 1.0 mM CdCl2 gradually increased during the culture period and the highest values of α, β and γ tocopherols (145.61, 25.52 and 18.56 μg/100 g) were detected in the cell cultures collected at day 6

  20. The effects of cadmium chloride on secondary metabolite production in Vitis vinifera cv. cell suspension cultures

    Directory of Open Access Journals (Sweden)

    Emine Sema Cetin

    2014-01-01

    Full Text Available BACKGROUND: Plant secondary metabolites are possess several biological activities such as anti-mutagenic, anti-carcinogenic, anti-aging, etc. Cell suspension culture is one of the most effective systems to produce secondary metabolites. It is possible to increase the phenolic compounds and tocopherols by using cell suspensions. Studies on tocopherols production by cell suspension cultures are seldom and generally focused on seed oil plants. Although fresh grape, grape seed, pomace and grape seed oil had tocopherols, with our best knowledge, there is no research on tocopherol accumulation in the grape cell suspension cultures. In this study, it was aimed to determine the effects of cadmium chloride treatments on secondary metabolite production in cell suspension cultures of grapevine. Cell suspensions initiated from callus belonging to petiole tissue was used as a plant material. Cadmium chloride was applied to cell suspension cultures in different concentration (1.0 mM and 1.5 mM to enhance secondary metabolite (total phenolics, total flavanols, total flavonols, trans-resveratrol, and α-, β-, γ- δ-tocopherols production. Cells were harvested at two days intervals until the 6th day of cultures. Amounts of total phenolics, total flavanols and total flavonols; trans-resveratrol and tocopherols (α-, β-, γ- and δ-tocopherols and dry cell weights were determined in the harvested cells. RESULTS: Phenolic contents were significantly affected by the sampling time and cadmium concentrations. The highest values of total phenolic (168.82 mg/100 g, total flavanol (15.94 mg/100 g, total flavonol (14.73 mg/100 g and trans-resveratrol (490.76 µg/100 g were found in cells treated with 1.0 mM CdCl2 and harvested at day 2. Contents of tocopherols in the cells cultured in the presence of 1.0 mM CdCl2 gradually increased during the culture period and the highest values of α, β and γ tocopherols (145.61, 25.52 and 18.56 µg/100 g were detected in the cell

  1. Monitoring Technology for Vehicle Loading Status Based on the Analysis of Suspension Vibration Characters

    Directory of Open Access Journals (Sweden)

    Shiwu Li

    2014-01-01

    Full Text Available Monitoring and early warning of vehicle risk status was one of the key technologies of transportation security, and real-time monitoring load status could reduce the transportation accidents effectively. In order to obtain vehicle load status information, vehicle characters of suspension were analyzed and simulated under different working conditions. On the basis of this, the device that can detect suspension load with overload protection structure was designed and a method of monitored vehicle load status was proposed. The monitoring platform of vehicle load status was constructed and developed for a FAW truck and system was tested on level-A road and body twist lane. The results show that the measurement error is less than 5% and horizontal centre-of-mass of vehicle was positioned accurately. The platform enables providing technical support for the real-time monitoring and warning of vehicles risk status in transit.

  2. Establishment of sorghum cell suspension culture system for ...

    African Journals Online (AJOL)

    Total soluble proteins (TSP) and culture filtrate (CF) proteins were extracted from the cell culture system and solubilised in urea buffer (9 M urea, 2 M thiourea and 4% CHAPS). Both onedimensional (1D) and two-dimensional (2D) gel analysis of these two proteomes show that the TSP and CF proteomes have different ...

  3. In vitro production of azadirachtin from cell suspension cultures of ...

    Indian Academy of Sciences (India)

    PRAKASH KUMAR G

    Tabata M, Mizukami H, Hiroaka N and Konoshima M 1974. Pigment formation in callus cultures of Lithospermum erythrorizon; Phytochemistry 13 927–932. Vander Esch S A, Giagnacovo G, Meccioni O and Vitali F 1993. Preliminary results on the production of azadirachtin by plant tissue culture of Azadirachta indica; G. Bot.

  4. An efficient recursive least square-based condition monitoring approach for a rail vehicle suspension system

    Science.gov (United States)

    Liu, X. Y.; Alfi, S.; Bruni, S.

    2016-06-01

    A model-based condition monitoring strategy for the railway vehicle suspension is proposed in this paper. This approach is based on recursive least square (RLS) algorithm focusing on the deterministic 'input-output' model. RLS has Kalman filtering feature and is able to identify the unknown parameters from a noisy dynamic system by memorising the correlation properties of variables. The identification of suspension parameter is achieved by machine learning of the relationship between excitation and response in a vehicle dynamic system. A fault detection method for the vertical primary suspension is illustrated as an instance of this condition monitoring scheme. Simulation results from the rail vehicle dynamics software 'ADTreS' are utilised as 'virtual measurements' considering a trailer car of Italian ETR500 high-speed train. The field test data from an E464 locomotive are also employed to validate the feasibility of this strategy for the real application. Results of the parameter identification performed indicate that estimated suspension parameters are consistent or approximate with the reference values. These results provide the supporting evidence that this fault diagnosis technique is capable of paving the way for the future vehicle condition monitoring system.

  5. Impact of stirred suspension bioreactor culture on the differentiation of murine embryonic stem cells into cardiomyocytes

    Directory of Open Access Journals (Sweden)

    Shafa Mehdi

    2011-12-01

    Full Text Available Abstract Background Embryonic stem cells (ESCs can proliferate endlessly and are able to differentiate into all cell lineages that make up the adult organism. Under particular in vitro culture conditions, ESCs can be expanded and induced to differentiate into cardiomyocytes in stirred suspension bioreactors (SSBs. However, in using these systems we must be cognizant of the mechanical forces acting upon the cells. The effect of mechanical forces and shear stress on ESC pluripotency and differentiation has yet to be clarified. The purpose of this study was to investigate the impact of the suspension culture environment on ESC pluripotency during cardiomyocyte differentiation. Results Murine D3-MHC-neor ESCs formed embyroid bodies (EBs and differentiated into cardiomyocytes over 25 days in static culture and suspension bioreactors. G418 (Geneticin was used in both systems from day 10 to enrich for cardiomyocytes by eliminating non-resistant, undifferentiated cells. Treatment of EBs with 1 mM ascorbic acid and 0.5% dimethyl sulfoxide from day 3 markedly increased the number of beating EBs, which displayed spontaneous and cadenced contractile beating on day 11 in the bioreactor. Our results showed that the bioreactor differentiated cells displayed the characteristics of fully functional cardiomyocytes. Remarkably, however, our results demonstrated that the bioreactor differentiated ESCs retained their ability to express pluripotency markers, to form ESC-like colonies, and to generate teratomas upon transplantation, whereas the cells differentiated in adherent culture lost these characteristics. Conclusions This study demonstrates that although cardiomyocyte differentiation can be achieved in stirred suspension bioreactors, the addition of medium enhancers is not adequate to force complete differentiation as fluid shear forces appear to maintain a subpopulation of cells in a transient pluripotent state. The development of successful ESC

  6. Differentiation of somatic embryos from protoplasts isolated from embryogenic suspension cultures of Abies alba L.

    Science.gov (United States)

    Hartmann, S; Lang, H; Reuther, G

    1992-10-01

    Embryogenic suspension cultures of Abies alba were established using an embryogenic suspensor mass culture originating from the zygotic embryo in immature seed explants (Schuller et al. 1989). Protoplasts were isolated from the suspension material. The protoplasts were immobilized in alginate layers in order to follow the development of single protoplasts. During the first days of protoplast culture a modified Kao and Michayluk (1975) medium proved to be necessary for subsequent divisions. The formation of proembryos succeeded within 2-3 weeks when subcultured with a modified Schenk and Hildebrandt (1972) liquid medium. Light, enhanced sugar concentration, and the addition of abscisic acid led to the formation of slightly green "torpedo"-shaped somatic embryos after 6-8 weeks from protoplast isolation.

  7. Effects of Selected Physicochemical Parameters on Zerumbone Production of Zingiber zerumbet Smith Cell Suspension Culture

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    Mahanom Jalil

    2015-01-01

    Full Text Available Zingiber zerumbet Smith is an important herb that contains bioactive phytomedicinal compound, zerumbone. To enhance cell growth and production of this useful compound, we investigated the growth conditions of cell suspension culture. Embryogenic callus generated from shoot bud was used to initiate cell suspension culture. The highest specific growth rate of cells was recorded when it was cultured in liquid Murashige and Skoog basal medium containing 3% sucrose with pH 5.7 and incubated under continuous shaking condition of 70 rpm for 16 h light and 8 h dark cycle at 24°C. Our results also revealed that the type of carbohydrate substrate, light regime, agitation speed, and incubation temperature could affect the production of zerumbone. Although the zerumbone produced in this study was not abundant compared to rhizome of Z. zerumbet, the possibility of producing zerumbone during early stage could serve as a model for subsequent improvement.

  8. Somatic Embryogenesis of Date Palm (Phoenix dactylifera L.) Through Cell Suspension Culture.

    Science.gov (United States)

    Naik, Poornananda M; Al-Khayri, Jameel M

    2016-01-01

    Date palm (Phoenix dactylifera L.) is the oldest and most economically important plant species distributed in the hot arid regions of the world. Propagation of date palm by seeds produces heterogeneous offspring with inferior field performance and poor fruit quality. Traditionally, date palm is propagated by offshoots, but this method is inefficient for mass propagation because of limited availability of offshoots. Plant regeneration through tissue culture is able to provide technologies for the large-scale propagation of healthy true-to-type plants. The most commonly used technology approach is somatic embryogenesis which presents a great potential for the rapid propagation and genetic resource preservation of this species. Significant progress has been made in the development and optimization of this regeneration pathway through the establishment of embryogenic suspension cultures. This chapter focuses on the methods employed for the induction of callus from shoot tip explants, establishment of cell suspension culture, and subsequent somatic embryogenesis and plant regeneration.

  9. Rheological characteristics of cell suspension and cell culture of Perilla frutescens.

    Science.gov (United States)

    Zhong, J J; Seki, T; Kinoshita, S; Yoshida, T

    1992-12-05

    Physical properties such as viscosity, fluid dynamic behavior of cell suspension, and size distribution of cell aggregates of a plant, Perilla frustescens, cultured in a liquid medium were studied. As a result of investigations using cells harvester after 12 days of cultivation in a flask, it was found that the apparent viscosity of the cell suspension did not change with any variation of cell concentration below 5 g dry cell/L but markedly increased when the cell concentration increased over 12.8 g dry cell/L. The cell suspension exhibited the characteristics of a Bingham plastic fluid with a small yield stress. The size of cell aggregates in the range 74 to 500 mum did not influence the rheological characteristics of the cell suspension. The rheological characteristics of cultivation mixtures of P. frutescens cultivated in a flask and in a bioreactor were also investigated. The results showed that the flow characteristics of the cell culture could be described by a Bingham plastic model. At the later stage of cultivation, the apparent viscosity increased steadily, even though the biomass concentration (by dry weight) decreased, due to the increase of individual cell size. (c) 1992 John Wiley & Sons, Inc.

  10. Evaluation of limonoid production in suspension cell culture of Citrus sinensis

    Directory of Open Access Journals (Sweden)

    Elisângela Fumagali Gerolino

    Full Text Available ABSTRACTThe use of cell and plant tissue culture techniques to produce economically important active metabolites has been growing. Among these substances are total limonoid aglycones, which are produced by "pera" orange (Citrus sinensis (L. Osbeck, Rutaceae and have received considerable attention because of their anticancer actions. The main objective of the present study was to analyze and compare the levels of limonoid aglycones in seeds, callus cultures (originating from seeds, callus cultures (originating from hypocotyls, cell suspensions from hypocotyls cells, and cell suspensions from cotyledons. The cell cultures or C. sinensis were obtained by inoculating two strains of callus in MS medium supplemented with 2.0 µM 2,4-dichlorophenoxyacetic acid, 7.0 µM benzyl aminopurine, and 3% (w/v sucrose in the dark. The highest concentrations of limonoid aglycone that were obtained were observed in cotyledon cell lines (240 mg/100 g dry weight that were produced on day 21 of culture and hypocotyl cell lines on day 7 (210 mg/100 g dry weight. Explants of different origins under the same culture conditions had different limonoid aglycone content. The present results may suggest strategies for enhancing the productivity of biologically important limonoid aglycones and investigating the complex pathways of these secondary metabolites in plant tissue cultures.

  11. Effects of auxins on growth and scopoletin accumulation in cell suspension cultures of Angelica archangelica L.

    Science.gov (United States)

    Siatka, T; Kasparová, M

    2008-01-01

    Scopoletin is a coumarin possessing many interesting biological effects, e.g., spasmolytic, anti-inflammatory, antimutagenic, antioxidant, antifungal, apoptosis-inducing, antiproliferative, acetylcholinesterase-inhibitory, and hypouricemic activities. Plant tissue cultures represent a promising alternative source of valuable plant-derived substances. A number of physical and chemical factors influence the cell growth and secondary metabolite biosynthesis in plant tissue cultures. The mechanism of their action is not completely understood. Besides other factors, plant growth regulators and light conditions play an important role. Effects of four auxins (2,4-dichlorophenoxyacetic acid, 2,4-D, alpha-naphthaleneacetic acid, NAA, beta-indoleacetic acid, IAA or beta-indolebutyric acid, IBA) at four concentrations (0.2, 2, 10 or 20 mg/l) on the culture growth and accumulation of scopoletin in the medium were tested in Angelica archangelica cell suspension cultures cultured under continuous light or in the dark. The highest culture growth was achieved with 2 mg/l 2,4-D, and 10 mg/l IAA. The best scopoletin levels were obtained with 0.2 mg/l 2,4-D, 2 mg/l 2,4-D, 10 mg/l NAA, and 20 mg/l IAA. The effects of light conditions were less marked than those of auxins and their concentrations in influencing both the cell growth and scopoletin accumulation in Angelica archangelica cell suspension cultures. The changes brought about by auxins were modified by light conditions.

  12. Non-linear dielectric monitoring of biological suspensions

    International Nuclear Information System (INIS)

    Treo, E F; Felice, C J

    2007-01-01

    Non-linear dielectric spectroscopy as a tool for in situ monitoring of enzyme assumes a non-linear behavior of the sample when a sinusoidal voltage is applied to it. Even many attempts have been made to improve the original experiments, all of them had limited success. In this paper we present upgrades made to a non-linear dielectric spectrometer developed and the results obtained when using different cells. We emphasized on the electrode surface, characterizing the grinding and polishing procedure. We found that the biological medium does not behave as expected, and the non-linear response is generated in the electrode-electrolyte interface. The electrochemistry of this interface can bias unpredictably the measured non-linear response

  13. Isolation of fatty acids and aromatics from cell suspension cultures of Lavandula angustifolia.

    Science.gov (United States)

    Topçu, Gülaçti; Herrmann, Gabriele; Kolak, Ufuk; Gören, C; Porzel, Andrea; Kutchan, Toni M

    2007-02-01

    Cell suspension cultures of Lavandula angustifolia Mill. ssp. angustifolia (syn.: L. officinalis Chaix.) afforded a fatty acid composition, cis and trans p-coumaric acids (=p-hydroxy cinnamic acids), and beta-sitosterol. The fatty acid composition was analyzed by GC-MS, and the structures of the isolated three compounds were determined by 1H- and 13C-NMR, and MS spectroscopic techniques.

  14. Effects of Growth Regulators on the Induction of Anthocyanin Synthesis in Carrot Suspension Cultures

    OpenAIRE

    Yoshihiro, Ozeki; Atsushi, Komamine; Department of Botany, Faculty of Science, The University of Tokyo:(Present)Department of Biology, College of Arts and Sciences, The University of Tokyo; Department of Botany, Faculty of Science, The University of Tokyo:(Present)Biological Institute, Faculty of Science, Tohoku University

    1986-01-01

    The effects of plant growth regulators were investigated on anthocyanin synthesis induced by removing auxin from carrot suspension cultures. Of the auxins tested, 2,4-D showed the strongest inhibiting effect on anthocyanin synthesis and had the strongest promoting effect on undifferentiated growth. When 2,4-D was added to anthocyanin synthesizing cells, in which cell division had ceased, anthocyanin synthesis was repressed immediately, accumulated anthocyanin disappeared and cell division res...

  15. Production of saponins from Panax ginseng suspension and adventitious root cultures

    Czech Academy of Sciences Publication Activity Database

    Langhansová, Lenka; Maršík, Petr; Vaněk, Tomáš

    2005-01-01

    Roč. 49, č. 3 (2005), s. 463-465 ISSN 0006-3134 R&D Projects: GA ČR(CZ) GP521/02/P064; GA MŠk(CZ) OC 843.10; GA MŠk(CZ) ME 671 Institutional research plan: CEZ:AV0Z4055905 Keywords : suspension cultures * bioreactor * ginsenosides Subject RIV: EI - Biotechnology ; Bionics Impact factor: 0.792, year: 2005

  16. Flavonoids and darkness lower PCD in senescing Vitis vinifera suspension cell cultures.

    Science.gov (United States)

    Bertolini, Alberto; Petrussa, Elisa; Patui, Sonia; Zancani, Marco; Peresson, Carlo; Casolo, Valentino; Vianello, Angelo; Braidot, Enrico

    2016-10-26

    Senescence is a key developmental process occurring during the life cycle of plants that can be induced also by environmental conditions, such as starvation and/or darkness. During senescence, strict control of genes regulates ordered degradation and dismantling events, the most remarkable of which are genetically programmed cell death (PCD) and, in most cases, an upregulation of flavonoid biosynthesis in the presence of light. Flavonoids are secondary metabolites that play multiple essential roles in development, reproduction and defence of plants, partly due to their well-known antioxidant properties, which could affect also the same cell death machinery. To understand further the effect of endogenously-produced flavonoids and their interplay with different environment (light or dark) conditions, two portions (red and green) of a senescing grapevine callus were used to obtain suspension cell cultures. Red Suspension cell Cultures (RSC) and Green Suspension cell Cultures (GSC) were finally grown under either dark or light conditions for 6 days. Darkness enhanced cell death (mainly necrosis) in suspension cell culture, when compared to those grown under light condition. Furthermore, RSC with high flavonoid content showed a higher viability compared to GSC and were more protected toward PCD, in accordance to their high content in flavonoids, which might quench ROS, thus limiting the relative signalling cascade. Conversely, PCD was mainly occurring in GSC and further increased by light, as it was shown by cytochrome c release and TUNEL assays. Endogenous flavonoids were shown to be good candidates for exploiting an efficient protection against oxidative stress and PCD induction. Light seemed to be an important environmental factor able to induce PCD, especially in GSC, which lacking of flavonoids were not capable of preventing oxidative damage and signalling leading to senescence.

  17. Neuropharmacological and neuroprotective activities of some metabolites produced by cell suspension culture of Waltheria americana Linn.

    Science.gov (United States)

    Mundo, Jorge; Villeda-Hernández, Juana; Herrera-Ruiz, Maribel; Gutiérrez, María Del Carmen; Arellano-García, Jesús; León-Rivera, Ismael; Perea-Arango, Irene

    2017-10-01

    Waltheria americana is a plant used in Mexican traditional medicine to treat some nervous system disorders. The aims of the present study were to isolate and determine the neuropharmacological and neurprotective activities of metabolites produced by a cell suspension culture of Waltheria americana. Submerged cultivation of W. americana cells provided biomass. A methanol-soluble extract (WAsc) was obtained from biomass. WAsc was fractionated yielding the chromatographic fractions 4WAsc-H 2 O and WAsc-CH 2 Cl 2 . For the determination of anticonvulsant activity in vivo, seizures were induced in mice by pentylenetetrazol (PTZ). Neuropharmacological activities (release of gamma amino butyric acid (GABA) and neuroprotection) of chromatographic fractions were determined by in vitro histological analysis of brain sections of mice post mortem. Fraction 4WAsc-H 2 O (containing saccharides) did not produce neuronal damage, neurodegeneration, interstitial tissue edema, astrocytic activation, nor cell death. Pretreatment of animals with 4WAsc-H 2 O and WAsc-CH 2 Cl 2 from W. americana cell suspensions induced an increase in: GABA release, seizure latency, survival time, neuroprotection, and a decrease in the degree of severity of tonic/tonic-clonic convulsions, preventing PTZ-induced death of up to 100% of animals of study. Bioactive compounds produced in suspension cell culture of W. americana produce neuroprotective and neuropharmacological activities associated with the GABAergic neurotransmission system. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  18. Quantitative proteome changes in Arabidopsis thaliana suspension-cultured cells in response to plant natriuretic peptides

    KAUST Repository

    Turek, Ilona

    2015-06-30

    Proteome changes in the Arabidopsis thaliana suspension cells in response to the A. thaliana plant natriuretic peptide (PNP), AtPNP-A (At2g18660) were assessed using quantitative proteomics employing tandem mass tag (TMT) labeling and tandem mass spectrometry (LC–MS/MS). In this study, we characterized temporal responses of suspension-cultured cells to 1 nM and 10 pM AtPNP-A at 0, 10 and 30 min post-treatment. Both concentrations we found to yield a distinct differential proteome signature. The data shown in this article are associated with the article “Plant natriuretic peptides induce a specific set of proteins diagnostic for an adaptive response to abiotic stress” by Turek et al. (Front. Plant Sci. 5 (2014) 661) and have been deposited to the ProteomeXchange with identifier PXD001386.

  19. Comparison of Cuminaldehyde Contents from Cell Suspension Cultures and Seeds of [Bunium persicum (Boiss. B. Fedtsch.

    Directory of Open Access Journals (Sweden)

    Sara KHOSRAVINIA

    2012-11-01

    Full Text Available The cell suspension culture and seed samples of Bunium persicum were extracted by supercritical fluid, hydrodistillation and solvent methods and analyzed by Gas Chromatography. In this study to compare the different methods of extractions, cuminaldehyde was targeted as one of the Black zira essential oil constitute. For callus induction the germinated seeds were cultured as explants on Murashige and Skoog medium supplemented with 2 mg/l 2,4-dichlorophenoxy acetic acid and 0.5 mg/l kinetin (treatment A as well as 2 mg/l ?-naphthalene acetic acid and 0.5 mg/l 6-benzyl aminopurine (treatment B and followed by cells suspension cultures establishment for the first time. The results of cell culture showed that cells from treatment B have a growth rate higher than A. All extracts were dissolved in 1 ml hexane and analyzed by Gas Chromatography. According to the Gas Chromatography analysis, cuminaldehyde was not detected in the supercritical fluid samples, while it was present in hydrodistillation and solvent extract. Cuminaldehyde percentage in cell and seed solvent extracts was 4.65% and 18.61% respectively. Gas Chromatography results also showed that no cuminaldehyde is present in media extracts, means no cuminaldehyde has been secreted into the medium.

  20. [Research on ursolic acid production of Eriobotrya japonica cell suspension culture in WAVE bioreactor].

    Science.gov (United States)

    Li, Hui-hua; Yao, De-heng; Xu, Jian; Wang, Wei; Chang, Qiang; Su, Ming-hua

    2015-05-01

    Through scale-up cultivation of Eriobotrya japonica suspension cells using WAVE bioreactor, the cell growth and ursolic acid (UA) accumulation were studied. The comparison test was carried out in the flask and the reactor with cell dry weight (DW) and UA content as evaluation indexes. The culture medium, DW and UA content were compared in 1 L and 5 L working volumes of bioreactor. The orthogonal test with main actors of inoculation amount, speed and angle of rotation was developed to find the optimal combination, in 1 L working volume of bioreactor. DW of the cell growth and the UA content in bioreactor were higher than those of the shaker by 105.5% and 27.65% respectively. In bioreactor, the dynamic changes of elements in the fluid culture, the dry weight of the cell growth and the UA content in 1 L and 5 L working volumes were similar. Inoculation of 80 g, rotational speed of 26 r · min(-1), and angle of 6 ° was the optimal combination, and the cell biomass of 19.01 g · L(-1) and the UA content of 27.750 mg · g(-1) were achieved after 100 h cultivation in 1 L working volume of bioreactor. WAVE Bioreactor is more suitable than flasks for the E. japonica cell suspension culture, and culture parameters can be achieved from 1 L to 5 L amplification.

  1. Cell line selection combined with jasmonic acid elicitation enhance camptothecin production in cell suspension cultures of Ophiorrhiza mungos L.

    Science.gov (United States)

    Deepthi, S; Satheeshkumar, K

    2017-01-01

    Ophiorrhiza mungos is a herbaceous medicinal plant which contains a quinoline alkaloid, camptothecin (CPT), an anticancer compound. A high-yielding cell line, O. mungos cell line-3 (OMC3) was selected from cell suspension cultures of O. mungos using cell aggregate cloning method and established cell suspension culture. OMC3 cell suspension produced significantly high biomass (9.25 ± 1.3 g/flask fresh weight (FW)) and CPT yield (0.095 ± 0.002 mg g -1 dry weight (DW)) compared with the original cell suspension. Inoculum size of OMC3 cell suspension culture was optimised as 14 g L -1 . Media optimisation has shown that 5 % (w/v) sucrose and an increased ammonium/nitrate concentration of 40/20 mM favoured CPT production, whereas 3 % (w/v) sucrose, an ammonium/nitrate concentration of 20/40 mM and 1.25 mM of phosphate favoured biomass accumulation. Jasmonic acid, chitin and salicylic acid was used to elicit CPT production in the original cell suspension culture and achieved significantly high CPT production with jasmonic acid (JA) elicitation. Further, OMC3 cell suspension culture was elicited with JA (50 μM) and obtained 1.12 ± 0.08 mg g -1 DW CPT and 9.52 ± 1.4 g/flask FW (190.4 g L -1 FW). The combination of cell line selection and elicitation has produced 18.66-fold increases in CPT production together with significantly high biomass yield. The study is helpful in the scale-up studies of O. mungos cell suspension culture in suitable bioreactor systems for the production of CPT.

  2. Real-time kinematic PPP GPS for structure monitoring applied on the Severn Suspension Bridge, UK

    Science.gov (United States)

    Tang, Xu; Roberts, Gethin Wyn; Li, Xingxing; Hancock, Craig Matthew

    2017-09-01

    GPS is widely used for monitoring large civil engineering structures in real time or near real time. In this paper the use of PPP GPS for monitoring large structures is investigated. The bridge deformation results estimated using double differenced measurements is used as the truth against which the performance of kinematic PPP in a real-time scenario for bridge monitoring is assessed. The towers' datasets with millimetre level movement and suspension cable dataset with centimetre/decimetre level movement were processed by both PPP and DD data processing methods. The consistency of tower PPP time series indicated that the wet tropospheric delay is the major obstacle for small deflection extraction. The results of suspension cable survey points indicate that an ionospheric-free linear measurement is competent for bridge deformation by PPP kinematic model, the frequency domain analysis yields very similar results using either PPP or DD. This gives evidence that PPP can be used as an alternative method to DD for large structure monitoring when DD is difficult or impossible because of large baseline lengths, power outages or natural disasters. The PPP residual tropospheric wet delays can be applied to improve the capacity of small movement extraction.

  3. Enhanced Production of Anthraquinones and Phenolic Compounds and Biological Activities in the Cell Suspension Cultures of Polygonum multiflorum

    Science.gov (United States)

    Thiruvengadam, Muthu; Rekha, Kaliyaperumal; Rajakumar, Govindasamy; Lee, Taek-Jun; Kim, Seung-Hyun; Chung, Ill-Min

    2016-01-01

    Anthraquinones (AQs) and phenolic compounds are important phytochemicals that are biosynthesized in cell suspension cultures of Polygonum multiflorum. We wanted to optimize the effects of plant growth regulators (PGRs), media, sucrose, l-glutamine, jasmonic acid (JA), and salicylic acid (SA) for the production of phytochemicals and biomass accumulation in a cell suspension culture of P. multiflorum. The medium containing Murashige and Skoog (MS) salts and 4% sucrose supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid, 0.5 mg/L thidiazuron, and 100 µM l-glutamine at 28 days of cell suspension culture was suitable for biomass accumulation and AQ production. Maximum biomass accumulation (12.5 and 12.35 g fresh mass (FM); 3 and 2.93 g dry mass (DM)) and AQ production (emodin 295.20 and 282 mg/g DM; physcion 421.55 and 410.25 mg/g DM) were observed using 100 µM JA and SA, respectively. JA- and SA-elicited cell cultures showed several-fold higher biomass accumulation and AQ production than the control cell cultures. Furthermore, the cell suspension cultures effectively produced 23 phenolic compounds, such as flavonols and hydroxycinnamic and hydroxybenzoic acid derivatives. PGR-, JA-, and SA-elicited cell cultures produced a higher amount of AQs and phenolic compounds. Because of these metabolic changes, the antioxidant, antimicrobial, and anticancer activities were high in the PGR-, JA-, and SA-elicited cell cultures. The results showed that the elicitors (JA and SA) induced the enhancement of biomass accumulation and phytochemical (AQs and phenolic compounds) production as well as biological activities in the cell suspension cultures of P. multiflorum. This optimized protocol can be developed for large-scale biomass accumulation and production of phytochemicals (AQs and phenolic compounds) from cell suspension cultures, and the phytochemicals can be used for various biological activities. PMID:27854330

  4. Enhanced Production of Anthraquinones and Phenolic Compounds and Biological Activities in the Cell Suspension Cultures of Polygonum multiflorum

    Directory of Open Access Journals (Sweden)

    Muthu Thiruvengadam

    2016-11-01

    Full Text Available Anthraquinones (AQs and phenolic compounds are important phytochemicals that are biosynthesized in cell suspension cultures of Polygonum multiflorum. We wanted to optimize the effects of plant growth regulators (PGRs, media, sucrose, l-glutamine, jasmonic acid (JA, and salicylic acid (SA for the production of phytochemicals and biomass accumulation in a cell suspension culture of P. multiflorum. The medium containing Murashige and Skoog (MS salts and 4% sucrose supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid, 0.5 mg/L thidiazuron, and 100 µM l-glutamine at 28 days of cell suspension culture was suitable for biomass accumulation and AQ production. Maximum biomass accumulation (12.5 and 12.35 g fresh mass (FM; 3 and 2.93 g dry mass (DM and AQ production (emodin 295.20 and 282 mg/g DM; physcion 421.55 and 410.25 mg/g DM were observed using 100 µM JA and SA, respectively. JA- and SA-elicited cell cultures showed several-fold higher biomass accumulation and AQ production than the control cell cultures. Furthermore, the cell suspension cultures effectively produced 23 phenolic compounds, such as flavonols and hydroxycinnamic and hydroxybenzoic acid derivatives. PGR-, JA-, and SA-elicited cell cultures produced a higher amount of AQs and phenolic compounds. Because of these metabolic changes, the antioxidant, antimicrobial, and anticancer activities were high in the PGR-, JA-, and SA-elicited cell cultures. The results showed that the elicitors (JA and SA induced the enhancement of biomass accumulation and phytochemical (AQs and phenolic compounds production as well as biological activities in the cell suspension cultures of P. multiflorum. This optimized protocol can be developed for large-scale biomass accumulation and production of phytochemicals (AQs and phenolic compounds from cell suspension cultures, and the phytochemicals can be used for various biological activities.

  5. Enhanced Production of Anthraquinones and Phenolic Compounds and Biological Activities in the Cell Suspension Cultures of Polygonum multiflorum.

    Science.gov (United States)

    Thiruvengadam, Muthu; Rekha, Kaliyaperumal; Rajakumar, Govindasamy; Lee, Taek-Jun; Kim, Seung-Hyun; Chung, Ill-Min

    2016-11-16

    Anthraquinones (AQs) and phenolic compounds are important phytochemicals that are biosynthesized in cell suspension cultures of Polygonum multiflorum . We wanted to optimize the effects of plant growth regulators (PGRs), media, sucrose, l-glutamine, jasmonic acid (JA), and salicylic acid (SA) for the production of phytochemicals and biomass accumulation in a cell suspension culture of P. multiflorum . The medium containing Murashige and Skoog (MS) salts and 4% sucrose supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid, 0.5 mg/L thidiazuron, and 100 µM l-glutamine at 28 days of cell suspension culture was suitable for biomass accumulation and AQ production. Maximum biomass accumulation (12.5 and 12.35 g fresh mass (FM); 3 and 2.93 g dry mass (DM)) and AQ production (emodin 295.20 and 282 mg/g DM; physcion 421.55 and 410.25 mg/g DM) were observed using 100 µM JA and SA, respectively. JA- and SA-elicited cell cultures showed several-fold higher biomass accumulation and AQ production than the control cell cultures. Furthermore, the cell suspension cultures effectively produced 23 phenolic compounds, such as flavonols and hydroxycinnamic and hydroxybenzoic acid derivatives. PGR-, JA-, and SA-elicited cell cultures produced a higher amount of AQs and phenolic compounds. Because of these metabolic changes, the antioxidant, antimicrobial, and anticancer activities were high in the PGR-, JA-, and SA-elicited cell cultures. The results showed that the elicitors (JA and SA) induced the enhancement of biomass accumulation and phytochemical (AQs and phenolic compounds) production as well as biological activities in the cell suspension cultures of P. multiflorum . This optimized protocol can be developed for large-scale biomass accumulation and production of phytochemicals (AQs and phenolic compounds) from cell suspension cultures, and the phytochemicals can be used for various biological activities.

  6. Accumulation of phenylpropanoid derivatives in chitosan-induced cell suspension culture of Cocos nucifera.

    Science.gov (United States)

    Chakraborty, Moumita; Karun, Anitha; Mitra, Adinpunya

    2009-01-01

    Chitosan-induced elicitation responses of dark-incubated Cocos nucifera (coconut) endosperm cell suspension cultures led to the rapid formation of phenylpropanoid derivatives, which essentially mimics the defense-induced biochemical changes in coconut palm as observed under in vivo conditions. An enhanced accumulation of p-hydroxybenzoic acid as the major wall-bound phenolics was evident. This was followed by p-coumaric acid and ferulic acid. Along with enhanced peroxidases activities in elicited lines, the increase in activities of the early phenylpropanoid pathway enzymes such as, phenylalanine ammonia lyase (PAL), p-coumaroyl-CoA ligase (4CL) and p-hydroxybenzaldehyde dehydrogenase (HBD) in elicited cell cultures were also observed. Furthermore, supplementation of specific inhibitors of PAL, C4H and 4CL in elicited cell cultures led to suppressed accumulation of p-hydroxybenzoic acid, which opens up interesting questions regarding the probable route of the biosynthesis of this phenolic acid in C. nucifera.

  7. Production of Monascus pigments as extracellular crystals by cell suspension culture.

    Science.gov (United States)

    Lu, Fengling; Liu, Lujie; Huang, Yaolin; Zhang, Xuehong; Wang, Zhilong

    2018-01-01

    It is generally accepted that Monascus pigments are predominantly cell-bound, including both intracellular and surface-bound pigments. This long-term misconception was corrected in the present work. Production of extracellular crystal pigments by submerged culture of Monascus sp. was confirmed by microscopic observation and collection of Monascus pigments from extracellular broth by direct membrane filtration. Following up the new fact, the bioactivity of mycelia as whole-cell biocatalyst for biosynthesis and biodegradation of Monascus pigments had been detailedly examined in both an aqueous solution and a nonionic surfactant micelle aqueous solution. Based on those experimental results, cell suspension culture in an aqueous medium was developed as a novel strategy for accumulation of high concentration of Monascus pigments. Thus, glucose feeding during submerged culture in the aqueous medium was carried out successfully and high orange Monascus pigments concentration of near 4 g/L was achieved.

  8. Effect of magnetic nanoparticles on tobacco BY-2 cell suspension culture.

    Science.gov (United States)

    Krystofova, Olga; Sochor, Jiri; Zitka, Ondrej; Babula, Petr; Kudrle, Vit; Adam, Vojtech; Kizek, Rene

    2012-12-20

    Nanomaterials are structures whose exceptionality is based on their large surface, which is closely connected with reactivity and modification possibilities. Due to these properties nanomaterials are used in textile industry (antibacterial textiles with silver nanoparticles), electronics (high-resolution imaging, logical circuits on the molecular level) and medicine. Medicine represents one of the most important fields of application of nanomaterials. They are investigated in connection with targeted therapy (infectious diseases, malignant diseases) or imaging (contrast agents). Nanomaterials including nanoparticles have a great application potential in the targeted transport of pharmaceuticals. However, there are some negative properties of nanoparticles, which must be carefully solved, as hydrophobic properties leading to instability in aqueous environment, and especially their possible toxicity. Data about toxicity of nanomaterials are still scarce. Due to this fact, in this work we focused on studying of the effect of magnetic nanoparticles (NPs) and modified magnetic nanoparticles (MNPs) on tobacco BY-2 plant cell suspension culture. We aimed at examining the effect of NPs and MNPs on growth, proteosynthesis - total protein content, thiols - reduced (GSH) and oxidized (GSSG) glutathione, phytochelatins PC2-5, glutathione S-transferase (GST) activity and antioxidant activity of BY-2 cells. Whereas the effect of NPs and MNPs on growth of cell suspension culture was only moderate, significant changes were detected in all other biochemical parameters. Significant changes in protein content, phytochelatins levels and GST activity were observed in BY-2 cells treated with MNPs nanoparticles treatment. Changes were also clearly evident in the case of application of NPs. Our results demonstrate the ability of MNPs to negatively affect metabolism and induce biosynthesis of protective compounds in a plant cell model represented by BY-2 cell suspension culture. The

  9. Photosynthetic carbon metabolism in photoautotrophic cell suspension cultures grown at low and high CO2

    International Nuclear Information System (INIS)

    Roeske, C.A.; Widholm, J.M.; Ogren, W.L.

    1989-01-01

    Photosynthetic carbon metabolism was characterized in four photoautotrophic cell suspension cultures. There was no apparent difference between two soybeans (Glycine max) and one cotton (Gossypium hirsutum) cell line which required 5% CO 2 for growth, and a unique cotton cell line that grows at ambient CO 2 (660 microliters per liter). Photosynthetic characteristics in all four lines were more like C 3 mesophyll leaf cells than the cell suspension cultures previously studied. The pattern of 14 C-labeling reflected the high ratio of ribulosebisphosphate carboxylase to phosphoenolpyruvate carboxylase activity and showed that CO 2 fixation occurred primarily by the C 3 pathway. Photorespiration occurred at 330 microliters per liter CO 2 , 21% O 2 as indicated by the synthesis of high levels of 14 C-labeled glycine and serine in a pulse-chase experiment and by oxygen inhibition of CO 2 fixation. Short-term CO 2 fixation in the presence and absence of carbonic anhydrase showed CO 2 , not HCO 3 - , to be the main source of inorganic carbon taken up by the low CO 2 -requiring cotton cells. The cells did not have a CO 2 -concentrating mechanism as indicated by silicone oil centrifugation experiments. Carbonic anhydrase was absent in the low CO 2 -requiring cotton cells, present in the high CO 2 -requiring soybean cell lines, and absent in other high CO 2 cell lines examined. Thus, the presence of carbonic anhydrase is not an essential requirement for photoautotrophy in cell suspension cultures which grow at either high or low CO 2 concentrations

  10. Effect of Magnetic Nanoparticles on Tobacco BY-2 Cell Suspension Culture

    Directory of Open Access Journals (Sweden)

    Rene Kizek

    2012-12-01

    Full Text Available Nanomaterials are structures whose exceptionality is based on their large surface, which is closely connected with reactivity and modification possibilities. Due to these properties nanomaterials are used in textile industry (antibacterial textiles with silver nanoparticles, electronics (high-resolution imaging, logical circuits on the molecular level and medicine. Medicine represents one of the most important fields of application of nanomaterials. They are investigated in connection with targeted therapy (infectious diseases, malignant diseases or imaging (contrast agents. Nanomaterials including nanoparticles have a great application potential in the targeted transport of pharmaceuticals. However, there are some negative properties of nanoparticles, which must be carefully solved, as hydrophobic properties leading to instability in aqueous environment, and especially their possible toxicity. Data about toxicity of nanomaterials are still scarce. Due to this fact, in this work we focused on studying of the effect of magnetic nanoparticles (NPs and modified magnetic nanoparticles (MNPs on tobacco BY-2 plant cell suspension culture. We aimed at examining the effect of NPs and MNPs on growth, proteosynthesis — total protein content, thiols — reduced (GSH and oxidized (GSSG glutathione, phytochelatins PC2-5, glutathione S-transferase (GST activity and antioxidant activity of BY-2 cells. Whereas the effect of NPs and MNPs on growth of cell suspension culture was only moderate, significant changes were detected in all other biochemical parameters. Significant changes in protein content, phytochelatins levels and GST activity were observed in BY-2 cells treated with MNPs nanoparticles treatment. Changes were also clearly evident in the case of application of NPs. Our results demonstrate the ability of MNPs to negatively affect metabolism and induce biosynthesis of protective compounds in a plant cell model represented by BY-2 cell suspension

  11. Saccharomyces cerevisiae gene expression changes during rotating wall vessel suspension culture

    Science.gov (United States)

    Johanson, Kelly; Allen, Patricia L.; Lewis, Fawn; Cubano, Luis A.; Hyman, Linda E.; Hammond, Timothy G.

    2002-01-01

    This study utilizes Saccharomyces cerevisiae to study genetic responses to suspension culture. The suspension culture system used in this study is the high-aspect-ratio vessel, one type of the rotating wall vessel, that provides a high rate of gas exchange necessary for rapidly dividing cells. Cells were grown in the high-aspect-ratio vessel, and DNA microarray and metabolic analyses were used to determine the resulting changes in yeast gene expression. A significant number of genes were found to be up- or downregulated by at least twofold as a result of rotational growth. By using Gibbs promoter alignment, clusters of genes were examined for promoter elements mediating these genetic changes. Candidate binding motifs similar to the Rap1p binding site and the stress-responsive element were identified in the promoter regions of differentially regulated genes. This study shows that, as in higher order organisms, S. cerevisiae changes gene expression in response to rotational culture and also provides clues for investigations into the signaling pathways involved in gravitational response.

  12. Biochemical precursor effects on the fatty acid production in cell suspension cultures of Theobroma cacao L.

    Science.gov (United States)

    Parra, O; Gallego, A M; Urrea, A; Rojas, L F; Correa, C; Atehortúa, L

    2017-02-01

    Cocoa butter (CB) is composed of 96% palmitic, stearic, oleic, linoleic and linolenic fatty acids that are responsible for the hardness, texture and fusion properties of chocolate. Through in vitro plant cell culture it is possible to modify CB lipid profiles and to study the fatty acid biosynthesis pathway on a subcellular level, evaluating fundamental aspects to enhance in vitro fatty acid production in a specific and controlled way. In this research, culture media was supplemented with acetate, biotin, pyruvate, bicarbonate and glycerol at three different concentrations and the effects on the biomass production (g/L), cell viability, and fatty acids profile and production was evaluated in in vitro cell suspensions culture. It was found that biotin stimulated fatty acid synthesis without altering cell viability and cell growth. It was also evident a change in the lipid profile of cell suspensions, increasing middle and long chain fatty acids proportion, which are unusual to those reported in seeds; thus implying that it is possible to modify lipid profiles according to the treatment used. According to the results of sucrose gradients and enzyme assays performed, it is proposed that cacao cells probably use the pentose phosphate pathway, mitochondria being the key organelle in the carbon flux for the synthesis of reductant power and fatty acid precursors. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  13. [The production of gastrodin through biotransformation of p-hydroxybenzaldehyde by cell suspension culture of Datura stramonium].

    Science.gov (United States)

    Gong, Jia-Shun; Ma, Wei-Peng; Pu, Jun-Xue; Xu, Shu-Guan; Zheng, Shuang-Qing; Xiao, Chun-Jie

    2006-10-01

    To investigate the production of p-hydroxymethylphenol-beta-D-glucoside (gastrodin) through biotransformation by plant cell suspension cultures. Using cell suspension cultures of Datura stramonium to convert the exogenous p-hydroxybenzaldehyde into gastrodin was conducted and the converted compounds were separated with a combination of multi-chromatography. Their chemical structures were determined on the basis of spectral analysis and chemical evidence. The conversion procedure of p-hydroxybenzaldehyde into gastrodin by Datura stramonium cell suspension cultures was established. The synthesized gastrodin (II) was isolated from the fermental liquor and identified by spectral analysis. At the same time, the p-hydroxybenzyl alcohol (I) converted through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of Datura stramonium was also isolated and identified. Two compounds were also isolated from the cell cultures and they were identified as beta-D-furanoallulose (III) and n-butyloxystyryl-beta-D-pyranoallulose (IV). Datura stramonium grown in suspension cultures can convert exogenous p-hydroxybenzaldehyde into the corresponding gastrodin.

  14. [Importance of 3T3 feeder layer to establish epithelial cultures from cell suspension obtained from corneo-scleral rims].

    Science.gov (United States)

    Cristovam, Priscila Cardoso; Glória, Maria Aparecida da; Melo, Gustavo Barreto; Gomes, José Alvaro Pereira

    2008-01-01

    To evaluate the importance of the presence of 3T3 fibroblasts for establishing limbal epithelial cultures from cell suspension obtained from corneo-scleral rims (CSR). Corneo-scleral rims from different donors (n=6) had their posterior stroma and endothelium stripped away. Each corneo-scleral rim was divided into three equal segments that were set up in tissue culture in three different conditions: one of the segments was placed with the epithelial side up on the bottom of a 6-well culture plate (Group A). The other two fragments were trypsinized and the obtained cell suspension was cultured with (Group B) or without (Group C) irradiaded 3T3 cells. The cells were cultured in supplemental hormonal epithelial medium (SHEM), the epithelial migration and clone formation in groups A, B and C were evaluated with phase contrast microscopy and rodamine B staining. Epithelial cell growth was observed in 4/6 rims (Group A). All epithelial cell suspensions that were cultured with 3T3 cells (Group B) formed clones. No adhesion or true clone formation (holo- or meroclones) was observed in the cell suspensions that were cultivated without 3T3 (Group C) (p=0.009). Epithelial cell suspension obtained from corneo-scleral rims in this model needs to be cultivated with 3T3 cells in order to form clones and establish limbal epithelial cell colonies with the potential to be used for ocular surface reconstruction.

  15. Elicitor-induced tyrosine decarboxylase in berberine-synthesizing suspension cultures of Thalictrum rugosum.

    Science.gov (United States)

    Gügler, K; Funk, C; Brodelius, P

    1988-01-04

    Tyrosine decarboxylase (EC 4.1.1.25) was induced in suspension cultures of Thalictrum rugosum by treatment with a yeast glucan elicitor. Maximum induction was observed at a carbohydrate concentration of 0.4 mg/g fresh weight of cells and maximum enzyme activity was reached 20 h after addition of elicitor. The enzyme was inducible in late exponential and early stationary growth phases. A good correlation between induced tyrosine decarboxylase activity and berberine biosynthesis has been established. It is suggested that tyrosine decarboxylase may be a key enzyme between primary and secondary metabolisms in the biosynthesis of norlaudanosoline-derived alkaloids.

  16. Proteomic characterization of golgi membranes enriched from Arabidopsis suspension cell cultures

    DEFF Research Database (Denmark)

    Hansen, Sara Fasmer; Ebert, Berit; Rautengarten, Carsten

    2016-01-01

    The plant Golgi apparatus has a central role in the secretory pathway and is the principal site within the cell for the assembly and processing of macromolecules. The stacked membrane structure of the Golgi apparatus along with its interactions with the cytoskeleton and endoplasmic reticulum has...... from an Arabidopsis cell suspension culture that can be used to investigate the proteome of this organelle. We also provide a useful workflow for the examination of proteomic data as the result of multiple analyses. Finally, we highlight a simple technique to validate the subcellular localization...

  17. Induced accumulation of oleanolic acid and ursolic acid in cell suspension cultures of Uncaria tomentosa.

    Science.gov (United States)

    Feria-Romero, Iris; Lazo, Elizabeth; Ponce-Noyola, Teresa; Cerda-García-Rojas, Carlos M; Ramos-Valdivia, Ana C

    2005-06-01

    Increasing sucrose from 20 to 50 g l(-1) in Uncaria tomentosa cell suspension cultures enhanced ursolic acid and oleanolic acid production from 129 +/- 61 to 553 +/- 193 microg g(-1) cell dry wt. The maximal concentration of both triterpenes (1680 +/- 39 microg g(-1) cell dry wt) was 8 days after elicitation by jasmonic acid, while yeast extract or citrus pectin treatments produced 1189 +/- 20 or 1120 +/- 26 microg g(-1) cell dry wt, respectively. The ratio of ursolic acid:oleanolic acid was constant at 70:30.

  18. [Effect of precursors on the production of coumarins in a suspension culture of Angelica archangelica L].

    Science.gov (United States)

    Siatka, T; Kasparová, M

    2002-01-01

    The effect of potential precursors (cinnamic acid, phenylalanine, tyrosine) in three concentrations (0.01; 0.1; 1 mmol/l) on the growth of the culture and coumarin production was investigated in the suspension culture of Angelica archangelica L. The cultures were cultivated under constant illumination (3500 lux) and in the dark. The growth of the culture in the dark was not affected by the precursors, cinnamic acid in a concentration of 1 mmol/l exerted toxic action. The growth of the culture cultivated under constant illumination was stimulated by tyrosine in a concentration of 0.1 and 1 mmol/l, and phenylalanine in a concentration of 0.1 mmol/l; phenylalanine and cinnamic acid in a concentration of 1 mmol/l inhibited the growth of the culture. In the cultivation in the dark, coumarin production was increased by phenylalanine in a concentration 0.01 mmol/l and by tyrosine in a concentration of 1 mmol/l. In the cultivation under constant illumination, coumarin production was decreased by the action of cinnamic acid, was not affected by tyrosine, and phenylalanine (0.01 and 0.1 mmol/l) increased it in comparison with the culture without precursors.

  19. [Fluid suspension co-culture of Dendrobium officinale protocorm and living fungus].

    Science.gov (United States)

    Meng, Zhixia; Shu, Ying; Wang, Chunlan; Guo, Shunxing

    2012-06-01

    To set up a system for fluid suspension co-culture of Dendrobium officinale protocorm and living fungus MF24, so as to provide certain scientific evidence for industrial production of protocorm. Whether the protocorm culture system was suitable for the normal growth of MF24 fungus were studied, the growth of protocorm cultured alone and co-cultured with the fungus were researched under light and dark culture conditions, the biomass and proliferation times were determined, and HPLC method was used to analyze and compare the changes of 11 characteristic peak areas in D. officinale protocorm. The MF24 fungus could grow normally in the 6,7-V liquid medium used to culture the protocorm, and when it was cultured by 8-10 hours per day under 1 500 lx, the growth rate of the fungus was slowed. Protocorm could grow normally in light and dark culture conditions, and add the MF24 fungus in the early cultivation stages of protocorm, both inhibit the growth of each other. In the protocorm for the growth stability to add 5 diameter 9 mm fungi block, the protocorm growth and chemical composition type had no significant effect. However, under illumination, co-cultured for 5 days protocorm of which 10 compounds content decreased 13.64% to 138.47%, in dark conditions, co-cultured for 5 days protocorm of which 7 compounds increased by 0.71% to 12.82%, and 4 compounds slightly reduced by 3.03% to 14. 14% compared with the control. Under the appropriate condition, living fungus MF24 could co-culture with the D. officinale protocorm, and affected the latter's secondary metabolite levels.

  20. Growth characteristics and nutrient depletion of Miscanthus x ogiformis Honda 'Giganteus' suspension cultures

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted

    1998-01-01

    The growth characteristics and nutrient depletion in suspension cultures of Miscanthus ogiformis Honda ‘Giganteus' grown in media containing either Murashige and Skoog or N6 basal nutrient salts were studied during a culture period of 15 days. Proline was added to both media in concentrations from...... 0 to 300 mM. The fresh and dry weights of the suspension aggregates and the concentrations of ammonium, nitrate, proline and sugar remaining in the medium were measured at different points in time during the culture period. The results showed an almost total depletion of ammonium but a limited...

  1. Evaluation of Antioxidant and Antibacterial Potentials of Nigella sativa L. Suspension Cultures under Elicitation

    Directory of Open Access Journals (Sweden)

    Hera Chaudhry

    2015-01-01

    Full Text Available Nigella sativa L. (family Ranunculaceae is an annual herb of immense medicinal properties because of its major active components (i.e., thymoquinone (TQ, thymohydroquinone (THQ, and thymol (THY. Plant tissue culture techniques like elicitation, Agrobacterium mediated transformation, hairy root culture, and so on, are applied for substantial metabolite production. This study enumerates the antibacterial and antioxidant potentials of N. sativa epicotyl suspension cultures under biotic and abiotic elicitation along with concentration optimization of the elicitors for enhanced TQ and THY production. Cultures under different concentrations of pectin and manganese chloride (MnCl2 elicitation (i.e., 5 mg/L, 10 mg/L, and 15 mg/L showed that the control, MnCl2 10 mg/L, and pectin 15 mg/L suspension extracts greatly inhibited the growth of E. coli, S. typhimurium, and S. aureus (MIC against E. coli, i.e., 2.35±0.8, 2.4±0.2, and 2.46±0.5, resp.. Elicitation decreased SOD enzyme activity whereas CAT enzyme activity increased remarkably under MnCl2 elicitation. MnCl2 10 mg/L and pectin 15 mg/L elicitation enhanced the DPPH radical inhibition ability, but ferric scavenging activity was comparable to the control. TQ and THY were quantified by LC-MS/MS in the cultures with high bioactive properties revealing maximum content under MnCl2 10 mg/L elicitation. Therefore, MnCl2 elicitation can be undertaken on large scale for sustainable metabolite production.

  2. Desiccation-Enhanced Maturation and Germination of Date Palm Somatic Embryos Derived from Cell Suspension Culture.

    Science.gov (United States)

    Boufis, Nazim; Titouh, Khayreddine; Khelifi, Lakhdar

    2017-01-01

    In vitro plant regeneration via somatic embryogenesis is a powerful tool for rapid, large-scale production of healthy true-to-type plants. This approach is suitable to preserve existing natural genetic variability and propagation of variability generated from genetic improvement programs, including crossing, somaclonal variation, mutagenesis, and somatic hybridization. This chapter outlines a simplified protocol for date palm regeneration via somatic embryogenesis induced in cell suspension cultures. In this protocol, culture medium composition is manipulated, including plant growth regulators and solid (addition of agar) and liquid media to achieve reduction of production cycle of somatic embryogenesis, which increases the multiplication rate of embryogenic callus and improves the quantity and quality of somatic embryos.

  3. Evaluation of Antioxidant Enzyme Activity and Biochemical Characterization from Static and Suspension Culture of Withania somnifera L.

    Directory of Open Access Journals (Sweden)

    Satyajit Kanungo

    2015-03-01

    Full Text Available Withania somnifera (L. Dunal, is an erect evergreen shrub commonly known as Ashwagandha. It is widely used in Ayurvedic and in the traditional pharmacopeia system of India. It is one of the major ingredients in many formulations prescribed for a variety of musculoskeletal conditions including arthritis and rheumatism. In the present study the variation in quality and quantity of protein and antioxidant enzymes were evaluated biochemically and enzymatically from the static and suspension cultures. The nodal segments had provided maximum callusing of 90.25±0.06 % with (1mg/l of BAP and Kn with (2mg/l of 2, 4-D. The static and suspension cultures were taken for the analysis of total soluble protein and screened for antioxidant enzyme activity [catalase (CAT, superoxide dismutase (SOD and guaiacol peroxidase (GPX]. The protein content (1.2016 µg/µl was found to be higher in static culture samples (0.870 µg/µl than the protein obtained from the suspension culture. The antioxidant enzyme activity (CAT, SOD and GPX was higher in static culture samples (301.01± 0.42, 198.92 ± 0.29, 103.75 ± 0.11 nkat/ mg of protein than that of suspension culture. Specific activity staining of isozyme pattern exhibited three isoforms (CAT 1, CAT 2 and CAT 3 in static culture samples but CAT 1 was absent in the sample extracted from suspension cultures.  In case of SOD, four bands (SOD 1, SOD 2, SOD 3 and SOD 4 were found in both the samples whereas intensity of GPX activity was found to be more in static culture but both the samples exhibited three isoforms such as  (GPX 1, GPX 2 and GPX 3. The supplementation of required nutrients along with the phytohormones under in vitro condition might be an enhancing factor to yield antioxidant enzymes in the static culture samples. 

  4. Production of triterpene acids by cell suspension cultures of Olea europaea.

    Science.gov (United States)

    Saimaru, Hiroshi; Orihara, Yutaka; Tansakul, Pimpimon; Kang, Young-Hwa; Shibuya, Masaaki; Ebizuka, Yutaka

    2007-05-01

    Olive (Olea europaea) contains large quantity of triterpene acids including oleanolic acid (6) as a major one. Varieties of biological activities exhibited by triterpene acids attracted our attentions, especially from pharmaceutical viewpoints. Cell culture of olive plant was induced and its triterpene constituents were studied. From the cell suspension cultures, six ursane type triterpene acids; ursolic acid (9), pomolic acid (10), rotundic acid (11), tormentic acid (12), 2alpha-hydroxyursolic acid (13) and 19alpha-hydroxyasiatic acid (14), and two oleanane type acids; oleanolic acid and maslinic acid (7), have been isolated. Quantity of ursane type triterpene acids produced by cell cultures was larger than that of oleanane type. Further, a multifunctional oxidosqualene cyclase (OSC) named OEA was cloned by homology based PCRs from the same cultured cells. Major product of OEA is alpha-amyrin (ursane skeleton), showing good accordance to higher content of ursane-type triterpene acids in the cultured cells, and strongly suggesting OEA to be a major contributor OSC for their production.

  5. More for less: Improving the biomass yield of a pear cell suspension culture by design of experiments.

    Science.gov (United States)

    Rasche, Stefan; Herwartz, Denise; Schuster, Flora; Jablonka, Natalia; Weber, Andrea; Fischer, Rainer; Schillberg, Stefan

    2016-03-18

    Plant cell suspension cultures are widely used for the production of recombinant proteins and secondary metabolites. One of the most important steps during process development is the optimization of yields by testing different cultivation parameters, including the components of the growth medium. However, we have shown that the biomass yield of a cell suspension culture derived from the pear cultivar Pyrus communis cv. Champagner Bratbirne can be significantly improved solely by varying the temperature, inoculum density, illumination, and incubation time. In contrast to medium optimization, these simple physical factors are easily controlled and varied, thereby reducing the effort required. Using an experimental design approach, we improved the biomass yield from 146 g fresh weight (FW)/L to 407 g FW/L in only 5 weeks, simultaneously reducing the costs of goods sold per kg biomass from € 125 to € 45. Our simple approach therefore offers a rapid, efficient and economical process for the optimization of plant cell suspension cultures.

  6. Single cell dual adherent-suspension co-culture micro-environment for studying tumor-stromal interactions with functionally selected cancer stem-like cells.

    Science.gov (United States)

    Chen, Yu-Chih; Zhang, Zhixiong; Fouladdel, Shamileh; Deol, Yadwinder; Ingram, Patrick N; McDermott, Sean P; Azizi, Ebrahim; Wicha, Max S; Yoon, Euisik

    2016-08-07

    Considerable evidence suggests that cancer stem-like cells (CSCs) are critical in tumor pathogenesis, but their rarity and transience has led to much controversy about their exact nature. Although CSCs can be functionally identified using dish-based tumorsphere assays, it is difficult to handle and monitor single cells in dish-based approaches; single cell-based microfluidic approaches offer better control and reliable single cell derived sphere formation. However, like normal stem cells, CSCs are heavily regulated by their microenvironment, requiring tumor-stromal interactions for tumorigenic and proliferative behaviors. To enable single cell derived tumorsphere formation within a stromal microenvironment, we present a dual adherent/suspension co-culture device, which combines a suspension environment for single-cell tumorsphere assays and an adherent environment for co-culturing stromal cells in close proximity by selectively patterning polyHEMA in indented microwells. By minimizing dead volume and improving cell capture efficiency, the presented platform allows for the use of small numbers of cells (<100 cells). As a proof of concept, we co-cultured single T47D (breast cancer) cells and primary cancer associated fibroblasts (CAF) on-chip for 14 days to monitor sphere formation and growth. Compared to mono-culture, co-cultured T47D have higher tumorigenic potential (sphere formation rate) and proliferation rates (larger sphere size). Furthermore, 96-multiplexed single-cell transcriptome analyses were performed to compare the gene expression of co-cultured and mono-cultured T47D cells. Phenotypic changes observed in co-culture correlated with expression changes in genes associated with proliferation, apoptotic suppression, tumorigenicity and even epithelial-to-mesechymal transition. Combining the presented platform with single cell transcriptome analysis, we successfully identified functional CSCs and investigated the phenotypic and transcriptome effects induced

  7. Comparisons of different measurements for monitoring diabetic cats treated with porcine insulin zinc suspension.

    Science.gov (United States)

    Martin, G J; Rand, J S

    2007-07-14

    Clinical measurements, including a subjective clinical score and water intake, and biochemical measurements, including blood glucose, fructosamine, beta-hydroxybutyrate, cholesterol, triglycerides, triglycerides corrected for free glycerol, glycerol and urine glucose were compared for monitoring diabetic cats treated with porcine insulin zinc suspension. The data were grouped by subjective clinical score and the sensitivity of each measurement in differentiating the grouped data was assessed. None of the measurements was able to differentiate between the ranked clinical score groups, but two-hourly measurements of blood glucose over 24 hours, water intake, urine glucose and fructosamine were useful in differentiating cats that subjectively had the water and food consumption and general appearance of a normal cat from cats in which the signs of diabetes were less well controlled. Measurements of plasma lipids were not well correlated with the other measurements. The measurements that were most closely correlated with apparently perfect clinical control were the J index, water intake and maximum and mean blood glucose concentrations. In practice, water intake, maximum blood glucose concentration, mean blood glucose concentration and urine glucose would be the most useful indicators of clinical control in diabetic cats treated with porcine insulin zinc suspension.

  8. pH and expansin action on single suspension-cultured tomato (Lycopersicon esculentum) cells.

    Science.gov (United States)

    Wang, C X; Wang, L; McQueen-Mason, S J; Pritchard, J; Thomas, C R

    2008-09-01

    The aim of this study was to measure key material properties of the cell walls of single suspension-cultured plant cells and relate these to cell-wall biochemistry. To this end, micromanipulation was used to compress single tomato cells between two flat surfaces until they ruptured, and force-deformation data were obtained. In addition to measuring the bursting force, we also determined the elastic (Young's) modulus of the cell walls by matching low strain (composites and proposed mechanisms of such failure. Through the measurement of cell-wall material properties using micromanipulation, it may be possible to understand more fully how cell-wall composition, structure and biochemistry lead to cell mechanical behaviour.

  9. [Effect of auxins on production of coumarin in a suspension culture of Angelica archangelica L].

    Science.gov (United States)

    Siatka, T; Kasparová, M

    2003-07-01

    The paper examined the effect of selected auxins (2,4-dichlorophenoxyacetic acid, alpha-naphthalene-acetic acid, beta-indoleacetic acid, beta-indoleburytic acid; each in four concentrations--0.2, 2, 10, and 20 mg/l) on the production of coumarins in the suspension culture of Angelica archangelica L. cultinated in the dark and under permanent lighting(3500 lux). The effect of the light regimen is, in comparison with auxins, less marked--the content of coumarins is mostly comparable both under permanent lighting and in the dark. The highest coumarin content was achieved with the use of alpha-naphthalene-acetic acid in a concentration of 0.2 mg/l with cultivation in the dark.

  10. Functional Thyroid Follicular Cells Differentiation from Human-Induced Pluripotent Stem Cells in Suspension Culture

    Directory of Open Access Journals (Sweden)

    Ayumi Arauchi

    2017-05-01

    Full Text Available The replacement of regenerated thyroid follicular cells (TFCs is a promising therapeutic strategy for patients with hypothyroidism. Here, we have succeeded in inducing functional TFCs from human-induced pluripotent stem cells (iPSCs in scalable suspension culture. Differentiation of iPSCs with Activin A treatment produced Sox17- and FoxA2-expressing definitive endodermal cells that also expressed thyroid transcription factors Pax8 and Nkx2-1. Further treatment with thyroid-stimulating hormone (TSH induced TFCs expressing various types of thyroid proteins including TSH receptor, sodium–iodide symporter, thyroglobulin, and thyroid peroxidase. Interestingly, differentiated cells secreted free thyroxine in vitro. These results indicate successful differentiation of human iPSCs to functional TFCs that may enable us to fabricate thyroid tissues for regenerative medicine and disease models.

  11. Statistical experimental designs for the production of secondary metabolites in plant cell suspension cultures.

    Science.gov (United States)

    Schmitz, Christian; Fritsch, Leonie; Fischer, Rainer; Schillberg, Stefan; Rasche, Stefan

    2016-12-01

    Statistical experimental designs, also known as the "design of experiments" (DoE) approach, are widely used to improve not only technical processes but also to answer questions in the agricultural, medical and social sciences. Although many articles have been published about the application of DoE in these fields, few studies have addressed the use of DoE in the plant sciences, particularly in the context of plant cell suspension cultures (PCSCs). Compounds derived from PCSCs can be developed as pharmaceuticals, chemical feedstocks and cosmetic ingredients, and statistical experimental designs can be used to improve the productivity of the cells and the yield and/or quality of the target compounds in a cost efficient manner. In this article, we summarize recent findings concerning the application of statistical approaches to improve the performance of PCSCs and discuss the potential future applications of this approach.

  12. Effects of oligosaccharides from endophytic Fusarium oxysporum Dzf17 on activities of defense-related enzymes in Dioscorea zingiberensis suspension cell and seedling cultures

    Directory of Open Access Journals (Sweden)

    Peiqin Li

    2014-07-01

    Conclusions: Both EOS and WOS significantly increased the activities of PAL, PPO and POD in the suspension cell and seedling cultures of D. zingiberensis. The results suggested that the oligosaccharides from the endophytic fungus F. oxysporum Dzf17 may be related to the activation and enhancement of the defensive mechanisms of D. zingiberensis suspension cell and seedling cultures.

  13. Elicitor-Induced l-Tyrosine Decarboxylase from Plant Cell Suspension Cultures : I. Induction and Purification.

    Science.gov (United States)

    Marques, I A; Brodelius, P E

    1988-09-01

    l-Tyrosine decarboxylase (EC 4.1.1.25) activity was induced in cell suspension cultures of Thalictrum rugosum Ait. and Eschscholtzia californica Cham. with a yeast polysaccharide preparation (elicitor). The highest l-tyrosine decarboxylase activity in extracts from 7-day-old cell cultures of E. californica was observed 5 hours after addition of 30 to 40 micrograms elicitor per gram cell fresh weight. The enzyme extracted from cells of E. californica was purified 1540-fold to a specific activity of 2.6 micromoles CO(2) produced per minute per milligram protein at pH 8.4 and 30 degrees C. Purified enzyme from T. rugosum showed a specific activity of 0.18 micromoles per minute per milligram protein. The purification procedure involved ammonium sulfate fractionation, anion-exchange fast protein liquid chromatography, ultrafiltration, and hydrophobic interaction chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the enzyme from the two plant cell cultures had subunits of identical molecular weight (56,300 +/- 300 daltons.

  14. Elicitor-Induced l-Tyrosine Decarboxylase from Plant Cell Suspension Cultures 1

    Science.gov (United States)

    Marques, Ivano A.; Brodelius, Peter E.

    1988-01-01

    l-Tyrosine decarboxylase (EC 4.1.1.25) activity was induced in cell suspension cultures of Thalictrum rugosum Ait. and Eschscholtzia californica Cham. with a yeast polysaccharide preparation (elicitor). The highest l-tyrosine decarboxylase activity in extracts from 7-day-old cell cultures of E. californica was observed 5 hours after addition of 30 to 40 micrograms elicitor per gram cell fresh weight. The enzyme extracted from cells of E. californica was purified 1540-fold to a specific activity of 2.6 micromoles CO2 produced per minute per milligram protein at pH 8.4 and 30°C. Purified enzyme from T. rugosum showed a specific activity of 0.18 micromoles per minute per milligram protein. The purification procedure involved ammonium sulfate fractionation, anion-exchange fast protein liquid chromatography, ultrafiltration, and hydrophobic interaction chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the enzyme from the two plant cell cultures had subunits of identical molecular weight (56,300 ± 300 daltons. Images Fig. 5 Fig. 6 PMID:16666277

  15. Biosynthesis of sterols and triterpenes in cell suspension cultures of Uncaria tomentosa.

    Science.gov (United States)

    Flores-Sánchez, Isvett J; Ortega-López, Jaime; del Carmen Montes-Horcasitas, María; Ramos-Valdivia, Ana C

    2002-12-01

    Pectin administered to Uncaria tomentosa cell suspension cultures, was found to increase the production of triterpene acids (ursolic and oleanolic acid), however, neither growth nor sterol accumulation were affected. Cell cultures showed that pectin treatment caused a rapid threefold increase in the activities of enzymes involved in the biosynthesis of C(5) and C(30 )isoprenoid, such as isopentenyl diphosphate isomerase and squalene synthase. The activity of a farnesyl diphosphatase, which could divert the flux of farnesyl diphosphate to farnesol, was two times lower in elicited than in control cells. Elicited cells also transformed more rapidly a higher percentage of [5-(3)H]mevalonic acid into triterpene acids. Interestingly, addition of terbinafine, an inhibitor of squalene epoxidase, to elicited cell cultures inhibited sterol accumulation while triterpene production was not inhibited. These results suggest that in U. tomentosa cells, both the previously mentioned enzymes and those involved in squalene 2,3-oxide formation play an important regulatory role in the biosynthesis of sterols and triterpenes.

  16. Dynamics of indole-3-acetic acid oxidase activity in suspension culture of sunflower crown-gall

    Directory of Open Access Journals (Sweden)

    Zofia Chirek

    2014-01-01

    Full Text Available IAA oxidase activity was determined in several growth phases of a suspension culture of sunflower crown-gall. During the short phase of intensive growth (zero passage - PO a negative correlation was noted between enzymatic activity and the rate of growth. IAA oxidase activity increased to a certain level is not a factor limiting cell division. For protraction of the phase of intensive growth (first passage - P1, however, a decrease in the activity of this enzyme seems indispensable. IAA oxidase activity in the tested culture is under the control of inhibitors present in the cells and medium. High enzyme inhibition was observed in PO cells during the phase, of intensive growth and in P1 at the beginning and in the middle part of this phase. These results suggest' that the -auxin level determined in earlier studies in sunflower crown-gall culture is controlled by the IAA oxidase set. During the long phase of intensive growth (P1 this control is of negative feedback type.

  17. [Formation of protodioscin and deltoside isomers in suspension cultures of Nepal yam (Dioscorea deltoidea Wall.) cells].

    Science.gov (United States)

    Khandy, M T; Titova, M V; Konstantinova, S V; Kochkin, D V; Ivanov, I M; Nosov, A M

    2016-01-01

    Changes in the content of the furostanol glycosides protodioscin and deltoside, particularly that of the (25S)-isomers of the glycosides, during suspension cultivation of different lines of Nepal yam (Dioscorea deltoidea Wall.) cells of the strain IFR-DM-0.5 has been investigated. The composition of furostanol glycosides has been characterized, and the dynamics of the accumulation of individual glycosides during lengthy subcultivation of cells maintained in flasks or in a barbotage bioreactor has been analyzed. A positive correlation between the growth and accumulation of substances that belonged to the class of furostanol glycosides has been demonstrated for cultured dioscorea cells, whereas the content of some of the individual glycosides varied considerably between the lines of the strain, cultures maintained under different conditions, and even between cells in different phases of the growth cycle. The increased content of (25R)-forms of the glycosides (protodioscin and deltoside) was correlated with a decrease in the cellular growth rate, whereas an increase in culture growth intensity occurred concomitantly to an increase of the amount of (25S)-isomers. This may be indicative of the specific stimulatory effect of (25S)-glycosides, but not the (25R)-forms, on cell proliferation in vitro. Thus, the concentration of (25S)-forms may increase due to the autoselection of cells capable of intensive division during prolonged cultivation.

  18. Feruloyl Oligosaccharides from Cell Walls of Suspension-Cultured Spinach Cells and Sugar Beet Pulp : STRUCTURE AND FUNCTION OF CELLS

    OpenAIRE

    Tadashi, ISHII; Forestry and Forest Products Research Institute

    1994-01-01

    Cell walls of suspension-cultured spinach cells and sugar beet pulp were separately hydrolyzed with Driselase. A feruloyl arabinobiose was isolated from both spinach cells and sugar beet. Four feruloyl oligosaccharides were obtained from sugar beet. The four oligosaccharides were characterized by NMR spectroscopy, methylation analysis and FAB-MS.

  19. Comparison of mesencephalic free-floating tissue culture grafts and cell suspension grafts in the 6-hydroxydopamine-lesioned rat

    DEFF Research Database (Denmark)

    Meyer, Morten; Widmer, H R; Wagner, B

    1998-01-01

    days in culture or directly as dissociated cell suspensions, and compared with regard to neuronal survival and ability to normalize rotational behavior in adult rats with unilateral 6-hydroxydopamine (6-OHDA) lesions. Other lesioned rats received injections of cell-free medium and served as controls...... of grafted dopaminergic neurons and to correlate that with the behavioral effects. Additional cultures and acutely prepared explants were also fixed and stored for histological investigation in order to estimate the loss of dopaminergic neurons in culture and after transplantation. Similar behavioral...... improvements in terms of significant reductions in amphetamine-induced rotations were observed in rats grafted with FFRT cultures (127%) and rats grafted with cell suspensions (122%), while control animals showed no normalization of rotational behavior. At 84 days after transplantation, there were similar...

  20. Induction of linalool as a pharmaceutical and medicinal metabolite via cell suspension culture of cumin (Cuminum cyminum L.).

    Science.gov (United States)

    Kazemi, N; Kahrizi, D; Mansouri, M; Karim, H; Vaziri, S; Zargooshi, J; Khanahmadi, M; Shokrinia, M; Mohammadi, N

    2016-05-30

    Cumin is an important medicinal plant in Iran. Plant cell suspension culture is a method for the production of medicinal and secondary metabolites. The linalool is a plant secondary metabolite that has been recognized as a neuroprotective agent. The purpose of this study was to evaluate the effects of salicylic acid elicitor on induction of linalool in cell suspension culture of cumin. For this purpose, the cumin seeds were prepared, to obtain sterile seedling, were disinfected with sodium hypochlorite and alcohol, and were cultured on MS basal medium. This research was conducted in two separate experiments including callus induction and suspension cultures. Leaf explants were prepared from sterile seedlings and used to produce callus on MS medium supplemented with 1 mg/l NAA and 0.5 mg/l BAP. In order to establish suspension culture, the appropriate calli were transferred to liquid medium. Then cell cultures were treated with elicitors. The effects of elicitor on the production of linalool secondary metabolite and cell viability were assessed by GC-Mass and tetrazolium test respectively. For this purpose, the salicylic acid (at concentrations of 0, 1, 2, 4 and 8 mg/l) was used. The experimental design was a completely randomized design with five treatments and three replications. The results of cell culture and GC-Mass analysis showed that salicylic acid had significant effects on the linalool production (linalool as a secondary metabolite and pharmaceutical agent in cell culture of cumin. It is necessary to determine the best combination of medium and elicitor.

  1. Monoterpenoid oxindole alkaloid production by Uncaria tomentosa (Willd) D.C. cell suspension cultures in a stirred tank bioreactor.

    Science.gov (United States)

    Trejo-Tapia, Gabriela; Cerda-García-Rojas, Carlos M; Rodríguez-Monroy, Mario; Ramos-Valdivia, Ana C

    2005-01-01

    Cell growth, monoterpenoid oxindole alkaloid (MOA) production, and morphological properties of Uncaria tomentosa cell suspension cultures in a 2-L stirred tank bioreactor were investigated. U. tomentosa (cell line green Uth-3) was able to grow in a stirred tank at an impeller tip speed of 95 cm/s (agitation speed of 400 rpm), showing a maximum biomass yield of 11.9 +/- 0.6 g DW/L and a specific growth rate of 0.102 d(-1). U. tomentosa cells growing in a stirred tank achieved maximum volumetric and specific MOA concentration (467.7 +/- 40.0 microg/L, 44.6 +/- 5.2 microg/g DW) at 16 days of culture. MOA chemical profile of cell suspension cultures growing in a stirred tank resembled that of the plant. Depending on culture time, from the total MOA produced, 37-100% was found in the medium in the bioreactor culture. MOA concentration achieved in a stirred tank was up to 10-fold higher than that obtained in Erlenmeyer flasks (agitated at 110 rpm). In a stirred tank, average area of the single cells of U. tomentosa increased up to 4-fold, and elliptical form factor increased from 1.40 to 2.55, indicating enlargement of U. tomentosa single cells. This work presents the first report of U. tomentosa green cell suspension cultures that grow and produce MOA in a stirred tank bioreactor.

  2. Metabolic labeling and membrane fractionation for comparative proteomic analysis of Arabidopsis thaliana suspension cell cultures.

    Science.gov (United States)

    Szymanski, Witold G; Kierszniowska, Sylwia; Schulze, Waltraud X

    2013-09-28

    Plasma membrane microdomains are features based on the physical properties of the lipid and sterol environment and have particular roles in signaling processes. Extracting sterol-enriched membrane microdomains from plant cells for proteomic analysis is a difficult task mainly due to multiple preparation steps and sources for contaminations from other cellular compartments. The plasma membrane constitutes only about 5-20% of all the membranes in a plant cell, and therefore isolation of highly purified plasma membrane fraction is challenging. A frequently used method involves aqueous two-phase partitioning in polyethylene glycol and dextran, which yields plasma membrane vesicles with a purity of 95% (1). Sterol-rich membrane microdomains within the plasma membrane are insoluble upon treatment with cold nonionic detergents at alkaline pH. This detergent-resistant membrane fraction can be separated from the bulk plasma membrane by ultracentrifugation in a sucrose gradient (2). Subsequently, proteins can be extracted from the low density band of the sucrose gradient by methanol/chloroform precipitation. Extracted protein will then be trypsin digested, desalted and finally analyzed by LC-MS/MS. Our extraction protocol for sterol-rich microdomains is optimized for the preparation of clean detergent-resistant membrane fractions from Arabidopsis thaliana cell cultures. We use full metabolic labeling of Arabidopsis thaliana suspension cell cultures with K(15)NO3 as the only nitrogen source for quantitative comparative proteomic studies following biological treatment of interest (3). By mixing equal ratios of labeled and unlabeled cell cultures for joint protein extraction the influence of preparation steps on final quantitative result is kept at a minimum. Also loss of material during extraction will affect both control and treatment samples in the same way, and therefore the ratio of light and heave peptide will remain constant. In the proposed method either labeled or

  3. Effects of mercury (II) species on cell suspension cultures of catharanthus roseus

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, L. (Hangzhou Univ. (China)); Cullen, W.R. (Univ. of British Columbia, Vancouver, British Columbia (Canada))

    1994-11-01

    Mercury has received considerable attention because of its high toxicity. Widespread contamination with mercury poses severe environmental problems despite our extensive knowledge of its toxicity in living systems. It is generally accepted that the toxicity of mercury is related to its oxidation states and species, the organic forms being more toxic than the inorganic forms. In the aquatic environment, the toxicity of mercury depends on the aqueous speciation of the mercuric ion (Hg[sup 2+]). Because of the complex coordination chemistry of mercury in aqueous systems, the nature of the Hg[sup 2+] species present in aquatic environments is influenced greatly by water chemistry (e. g, pH, inorganic ion composition, and dissolved organics). Consequently, the influence of environmental factors on the aqueous speciation of mercury has been the focus of much attention. However, there is very little information available regarding the effects of the species and speciation on Hg (II) toxicity in plant-tissue cultures. Catharanthus roseus (C. roseus), commonly called the Madagascar Periwinkle, is a member of the alkaloid rich family Apocynaceae. The present investigation was concerned with the toxicity of mercury on the growth of C. roseus cell suspension cultures as influenced by mercury (II) species and speciation. The specific objectives of the study were to (a) study the effects of mercury species on the growth of C. roseus cultures from the point of view of environmental biology and toxicology; (b) evaluate the effects of selenate, selenite and selected ligands such as chloride, 1-cysteine in the media on the acute toxicity of mercuric oxide; (c) determine the impact of the initial pH of the culture media on the toxicities of mercuric compounds; (d) discuss the dependence of the toxicity on the chemical species and speciation of Hg (II). 11 refs., 7 figs., 2 tabs.

  4. Inverse problem analysis of pluripotent stem cell aggregation dynamics in stirred-suspension cultures

    Science.gov (United States)

    Rostami, Mahboubeh Rahmati; Wu, Jincheng; Tzanakakis, Emmanuel S.

    2015-01-01

    The cultivation of stem cells as aggregates in scalable bioreactor cultures is an appealing modality for the large-scale manufacturing of stem cell products. Aggregation phenomena are central to such bioprocesses affecting the viability, proliferation and differentiation trajectory of stem cells but a quantitative framework is currently lacking. A population balance equation (PBE) model was used to describe the temporal evolution of the embryonic stem cell (ESC) cluster size distribution by considering collision-induced aggregation and cell proliferation in a stirred-suspension vessel. For ESC cultures at different agitation rates, the aggregation kernel representing the aggregation dynamics was successfully recovered as a solution of the inverse problem. The rate of change of the average aggregate size was greater at the intermediate rate tested suggesting a trade-off between increased collisions and agitation-induced shear. Results from forward simulation with obtained aggregation kernels were in agreement with transient aggregate size data from experiments. We conclude that the framework presented here can complement mechanistic studies offering insights into relevant stem cell clustering processes. More importantly from a process development standpoint, this strategy can be employed in the design and control of bioreactors for the generation of stem cell derivatives for drug screening, tissue engineering and regenerative medicine. PMID:26036699

  5. Biosynthesis of camalexin from tryptophan pathway intermediates in cell-suspension cultures of Arabidopsis.

    Science.gov (United States)

    Zook, M

    1998-12-01

    Camalexin (3-thiazol-2'-yl-indole) is the principal phytoalexin that accumulates in Arabidopsis after infection by fungi or bacteria. Camalexin accumulation was detectable in Arabidopsis cell-suspension cultures 3 to 5 h after inoculation with Cochliobolus carbonum (Race 1), and then increased rapidly from 7 to 24 h after inoculation. Levels of radioactivity incorporated into camalexin during a 1.5-h pulse labeling with [14C]anthranilate also increased with time after fungal inoculation. The levels of radioactive incorporation into camalexin increased rapidly between 7 and 18 h after inoculation, and then decreased along with camalexin accumulation. Relatively low levels of radioactivity from [14C]anthranilate incorporated into camalexin in the noninoculated controls. Autoradiographic analysis of the accumulation of chloroform-extractable metabolites labeled with [14C]anthranilate revealed a transient increase in the incorporation of radioactivity into indole in fungus-inoculated Arabidopsis cell cultures. The time-course measurement of radioactive incorporation into camalexin during a 1.5-h pulse labeling with [14C]indole was similar to that with [14C]anthranilate. These data suggest that indole destined for camalexin synthesis is produced by a separate enzymatic reaction that does not involve tryptophan synthase.

  6. Biosynthesis of Camalexin from Tryptophan Pathway Intermediates in Cell-Suspension Cultures of Arabidopsis1

    Science.gov (United States)

    Zook, Michael

    1998-01-01

    Camalexin (3-thiazol-2′-yl-indole) is the principal phytoalexin that accumulates in Arabidopsis after infection by fungi or bacteria. Camalexin accumulation was detectable in Arabidopsis cell-suspension cultures 3 to 5 h after inoculation with Cochliobolus carbonum (Race 1), and then increased rapidly from 7 to 24 h after inoculation. Levels of radioactivity incorporated into camalexin during a 1.5-h pulse labeling with [14C]anthranilate also increased with time after fungal inoculation. The levels of radioactive incorporation into camalexin increased rapidly between 7 and 18 h after inoculation, and then decreased along with camalexin accumulation. Relatively low levels of radioactivity from [14C]anthranilate incorporated into camalexin in the noninoculated controls. Autoradiographic analysis of the accumulation of chloroform-extractable metabolites labeled with [14C]anthranilate revealed a transient increase in the incorporation of radioactivity into indole in fungus-inoculated Arabidopsis cell cultures. The time-course measurement of radioactive incorporation into camalexin during a 1.5-h pulse labeling with [14C]indole was similar to that with [14C]anthranilate. These data suggest that indole destined for camalexin synthesis is produced by a separate enzymatic reaction that does not involve tryptophan synthase. PMID:9847113

  7. Design of serum-free medium for suspension culture of CHO cells on the basis of general commercial media.

    Science.gov (United States)

    Miki, Hideo; Takagi, Mutsumi

    2015-08-01

    The design of serum-free media for suspension culture of genetically engineered Chinese hamster ovary (CHO) cells using general commercial media as a basis was investigated. Subcultivation using a commercial serum-free medium containing insulin-like growth factor (IGF)-1 with or without FCS necessitated additives other than IGF-1 to compensate for the lack of FCS and improve cell growth. Suspension culture with media containing several combinations of growth factors suggested the effectiveness of addition of both IGF-1 and the lipid signaling molecule lysophosphatidic acid (LPA) for promoting cell growth. Subcultivation of CHO cells in suspension culture using the commercial serum-free medium EX-CELL™302, which contained an IGF-1 analog, supplemented with LPA resulted in gradually increasing specific growth rate comparable to the serum-containing medium and in almost the same high antibody production regardless of the number of generations. The culture with EX-CELL™302 supplemented with LPA in a jar fermentor with pH control at 6.9 showed an apparently higher cell growth rate than the cultures without pH control and with pH control at 6.8. The cell growth in the medium supplemented with aurintricarboxylic acid (ATA), which was much cheaper than IGF-1, in combination with LPA was synergistically promoted similarly to that in the medium supplemented with IGF-1 and LPA. In conclusion, the serum-free medium designed on the basis of general commercial media could support the growth of CHO cells and antibody production comparable to serum-containing medium in suspension culture. Moreover, the possibility of cost reduction by the substitution of IGF-1 with ATA was also shown.

  8. Enhanced Biosynthesis of Withanolides by Elicitation and Precursor Feeding in Cell Suspension Culture of Withania somnifera (L.) Dunal in Shake-Flask Culture and Bioreactor

    OpenAIRE

    Sivanandhan, Ganeshan; Selvaraj, Natesan; Ganapathi, Andy; Manickavasagam, Markandan

    2014-01-01

    The present study investigated the biosynthesis of major and minor withanolides of Withania somnifera in cell suspension culture using shake-flask culture and bioreactor by exploiting elicitation and precursor feeding strategies. Elicitors like cadmium chloride, aluminium chloride and chitosan, precursors such as cholesterol, mevalonic acid and squalene were examined. Maximum total withanolides detected [withanolide A (7606.75 mg), withanolide B (4826.05 mg), withaferin A (3732.81 mg), withan...

  9. Influence of auxins and sucrose in monoterpenoid oxindole alkaloid production by Uncaria tomentosa cell suspension cultures.

    Science.gov (United States)

    Luna-Palencia, Gabriela R; Cerda-García-Rojas, Carlos M; Rodríguez-Monroy, Mario; Ramos-Valdivia, Ana C

    2005-01-01

    Growth and alkaloid production in Uncaria tomentosa cell suspension cultures were studied in Murashige and Skoog medium supplemented with 10 microM 2,4-dichlorophenoxyacetic acid, 10 microM kinetin, and 58 mM sucrose for maintenance and with 10 microM indole-3-acetic acid, 10 microM kinetin, and 58 mM sucrose for production. A U. tomentosa pale Uth-3 cell line, cultured in the production medium, showed a reduced lag phase and a specific growth rate (mu) of 0.27 day(-1), while cells growing in the maintenance medium showed mu = 0.20 day(-1). U. tomentosa cells growing in the production medium produced monoterpenoid oxindole alkaloids (MOA) in amounts of 10.2 +/- 1.6 microg g(-1) dry weight (DW). The chemical profile of MOA produced by in vitro cell cultures was similar to that found in the plant. After 10 subcultures, maximum MOA production decreased to 2.0 +/- 0.7 microg g(-1) DW, while tryptamine alkaloids (TA) were produced with a maximum of 6.2 +/- 0.4 microg g(-1) DW. The increase of initial sucrose concentration up to 145 mM in the production medium enhanced the cell biomass by 3.2-fold (from 10.2 +/- 0.1 to 32.8 +/- 1.1 g DW L(-1)), reduced mu from 0.27 to 0.23 day(-1), and provoked a substantial accumulation of TA (23.1 +/- 4.7 microg g(-1) DW). A high sucrose concentration stimulated MOA production in the maintenance medium (2.7 +/- 0.5 microg g(-1) DW), even in the presence of 2,4-dichlorophenoxyacetic acid.

  10. Long-Term In-Service Monitoring and Performance Assessment of the Main Cables of Long-Span Suspension Bridges.

    Science.gov (United States)

    Deng, Yang; Liu, Yang; Chen, Suren

    2017-06-16

    Despite the recent developments in structural health monitoring, there remain great challenges for accurately, conveniently, and economically assessing the in-service performance of the main cables for long-span suspension bridges. A long-term structural health monitoring technique is developed to measure the tension force with a conventional sensing technology and further provide the in-service performance assessment strategy of the main cable. The monitoring system adopts conventional vibrating strings transducers to monitor the tension forces of separate cable strands of the main cable in the anchor span. The performance evaluation of the main cable is conducted based on the collected health monitoring data: (1) the measured strand forces are used to derive the overall tension force of a main cable, which is further translated into load bearing capacity assessment using the concept of safety factor; and (2) the proposed technique can also evaluate the uniformity of tension forces from different cable strands. The assessment of uniformity of strand forces of a main cable offers critical information in terms of potential risks of partial damage and performance deterioration of the main cable. The results suggest the proposed low-cost monitoring system is an option to provide approximate estimation of tension forces of main cables for suspension bridges. With the long-term monitoring data, the proposed monitoring-based evaluation methods can further provide critical information to assess the safety and serviceability performance of main cables.

  11. Efficient Secretion of Recombinant Proteins from Rice Suspension-Cultured Cells Modulated by the Choice of Signal Peptide

    Science.gov (United States)

    Huang, Li-Fen; Tan, Chia-Chun; Yeh, Ju-Fang; Liu, Hsin-Yi; Liu, Yu-Kuo; Ho, Shin-Lon; Lu, Chung-An

    2015-01-01

    Plant-based expression systems have emerged as a competitive platform in the large-scale production of recombinant proteins. By adding a signal peptide, αAmy3sp, the desired recombinant proteins can be secreted outside transgenic rice cells, making them easy to harvest. In this work, to improve the secretion efficiency of recombinant proteins in rice expression systems, various signal peptides including αAmy3sp, CIN1sp, and 33KDsp have been fused to the N-terminus of green fluorescent protein (GFP) and introduced into rice cells to explore the efficiency of secretion of foreign proteins. 33KDsp had better efficiency than αAmy3sp and CIN1sp for the secretion of GFP from calli and suspension-cultured cells. 33KDsp was further applied for the secretion of mouse granulocyte-macrophage colony-stimulating factor (mGM-CSF) from transgenic rice suspension-cultured cells; approximately 76%–92% of total rice-derived mGM-CSF (rmGM-CSF) was detected in the culture medium. The rmGM-CSF was bioactive and could stimulate the proliferation of a murine myeloblastic leukemia cell line, NSF-60. The extracellular yield of rmGM-CSF reached 31.7 mg/L. Our study indicates that 33KDsp is better at promoting the secretion of recombinant proteins in rice suspension-cultured cell systems than the commonly used αAmy3sp. PMID:26473722

  12. Biomass Yield and Steviol Glycoside Production in Callus and Suspension Culture of Stevia rebaudiana Treated with Proline and Polyethylene Glycol.

    Science.gov (United States)

    Gupta, Pratibha; Sharma, Satyawati; Saxena, Sanjay

    2015-06-01

    Enhanced production of steviol glycosides (SGs) was observed in callus and suspension culture of Stevia rebaudiana treated with proline and polyethylene glycol (PEG). To study their effect, yellow-green and compact calli obtained from in vitro raised Stevia leaves were sub-cultured on MS medium supplemented with 2.0 mg l(-1) NAA and different concentrations of proline (2.5-10 mM) and PEG (2.5-10 %) for 2 weeks, and incubated at 24 ± 1 °C and 22.4 μmol m(-2) s(-1) light intensity provided by white fluorescent tubes for 16 h. Callus and suspension culture biomass (i.e. both fresh and dry weight content) was increased with 5 mM proline and 5 % PEG, while at further higher concentrations, they got reduced. Further, quantification of SGs content in callus (collected at 15th day) and suspension culture (collected at 10th and 15th day) treated with and without elicitors was analysed by HPLC. It was observed that chemical stress enhanced the production of SGs significantly. In callus, the content of SGs increased from 0.27 (control) to 1.09 and 1.83 % with 7.5 mM proline and 5 % PEG, respectively, which was about 4.0 and 7.0 times higher than control. However, in the case of suspension culture, the same concentrations of proline and polyethylene glycol enhanced the SG content from 1.36 (control) to 5.03 and 6.38 %, respectively, on 10th day which were 3.7 times and 4.7 times higher than control.

  13. Elicitor-Induced l-Tyrosine Decarboxylase from Plant Cell Suspension Cultures : II. Partial Characterization.

    Science.gov (United States)

    Marques, I A; Brodelius, P E

    1988-09-01

    Properties of purified l-tyrosine decarboxylase (EC 4.1.1.25) from elicitor-induced cell suspension cultures of Eschscholtzia californica Cham. and Thalictrum rugosum Ait. are described. l-Tyrosine decarboxylase is a dimeric enzyme with a molecular weight of 112,600 +/- 600 daltons. The isoelectric point was estimated to be at pH 5.2 and pH 5.4 for the enzyme from E. californica and T. rugosum, respectively. The purified enzymes were stabilized in the presence of pyridoxal-5-phosphate. Optimum pH for the enzyme from both plants was found to be 8.4. Enzyme activity was dependent on exogeneously supplied pyridoxal-5-phosphate. The enzyme decarboxylated l-tyrosine and l-beta-3,4-dihydroxyphenylalanine but was inactive toward l-phenylalanine and l-tryptophan. Apparent K(m) values of Eschscholtzia- and Thalictrum-decarboxylase for l-tyrosine were 0.25 +/- 0.03 and 0.27 +/- 0.04 millimolar, respectively. Similar affinities were found for l-3,4-dihydroxyphenylalanine. Eschscholtzial-tyrosine decarboxylase was strongly inhibited by the phenylalanine analogue l-alpha-aminooxy-beta-phenylpropionate and largely unaffected by d,l-alpha-monofluoromethyl-3,4-dihydroxyphenylalanine and alpha-difluoromethyltyrosine.

  14. Phenylpropanoid Metabolism in Suspension Cultures of Vanilla planifolia Andr. : IV. Induction of Vanillic Acid Formation.

    Science.gov (United States)

    Funk, C; Brodelius, P E

    1992-05-01

    Kinetin is used as an elicitor to induce vanillic acid formation in cell suspension cultures of Vanilla planifolia. Maximal induction is observed at a kinetin concentration of 20 micrograms per gram of fresh weight of cells. Vanillic acid synthesis is observed a few hours after elicitation. The effects of kinetin on the activity of some enzymes of the phenylpropanoid pathway, i.e. phenylalanine ammonia-lyase, 4-hydroxycinnamate:coenzyme A ligase and uridine 5'-diphosphate-glucose:trans-cinnamic acid glucosyltransferase, are reported and compared to the effects of chitosan. The former two enzymes are induced by chitosan with a maximum activity of approximately 25 to 40 hours after elicitation. All three enzymes are induced by kinetin with maximum activities for phenylalanine ammonia lyase and 4-hydroxycinnamate:coenzyme A ligase at approximately 50 hours after induction, whereas maximum glucosyltransferase activity is seen already after 24 hours. Furthermore, both elicitors induced the formation of lignin-like material, whereas only kinetin induced vanillic acid biosynthesis. Finally, kinetin but not chitosan induces catechol-4-O-methyltransferase activity, catalyzing the formation of 4-methoxycinnamic acids, which were shown to be intermediates of hydroxybenzoic acid biosynthesis within cells of V. planifolia. It is suggested that this methyltransferase is directly involved in the biosynthesis of vanillic acid.

  15. Treatment strategies for high resveratrol induction in Vitis vinifera L. cell suspension culture

    Directory of Open Access Journals (Sweden)

    Thu V. Vuong

    2014-06-01

    Full Text Available Bioprocesses capable of producing large scales of resveratrol at nutraceutical grade are in demand. This study herein investigated treatment strategies to induce the production of resveratrol in Vitis vinifera L. cell suspension cultures. Among seven investigated elicitors, jasmonic acid (JA, salicylic acid, β-glucan (GLU, and chitosan enhanced the production of intracellular resveratrol manyfold. The combined treatment of JA and GLU increased extracellular resveratrol production by up to tenfold. The application of Amberlite XAD-7 resin for in situ removal and artificial storage of secreted resveratrol further increased resveratrol production by up to four orders of magnitude. The level of resveratrol produced in response to the combined treatment with 200 g/L XAD-7, 10 μM JA and 1 mg/mL GLU was approximately 2400 mg/L, allowing the production of resveratrol at an industrial scale. The high yield of resveratrol is due to the involvement of a number of mechanisms working in concert.

  16. Agrobacterium-mediated transformation of Vitis Cv. Monastrell suspension-cultured cells: Determination of critical parameters.

    Science.gov (United States)

    Chu, Mingyu; Quiñonero, Carmen; Akdemir, Hülya; Alburquerque, Nuria; Pedreño, María Ángeles; Burgos, Lorenzo

    2016-05-01

    Although some works have explored the transformation of differentiated, embryogenic suspension-cultured cells (SCC) to produce transgenic grapevine plants, to our knowledge this is one of the first reports on the efficient transformation of dedifferentiated Vitis vinifera cv Monastrell SCC. This protocol has been developed using the sonication-assisted Agrobacterium-mediated transformation (SAAT) method. A construct harboring the selectable nptII and the eyfp/IV2 marker genes was used in the study and transformation efficiencies reached over 50 independent transformed SCC per gram of infected cells. Best results were obtained when cells were infected at the exponential phase. A high density plating (500 mg/dish) gave significantly better results. As selective agent, kanamycin was inefficient for the selection of Monastrell transformed SCC since wild type cells were almost insensitive to this antibiotic whereas application of paromomycin resulted in very effective selection. Selected eyfp-expressing microcalli were grown until enough tissue was available to scale up a new transgenic SCC. These transgenic SCC lines were evaluated molecularly and phenotypically demonstrating the presence and integration of both transgenes, the absence of Agrobacterium contamination and the ability of the transformed SCC to grow in highly selective liquid medium. The methodology described here opens the possibility of improving the production of valuable metabolites. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:725-734, 2016. © 2016 American Institute of Chemical Engineers.

  17. Affinity Purification and Characterization of Functional Tubulin from Cell Suspension Cultures of Arabidopsis and Tobacco1

    Science.gov (United States)

    Fujita, Satoshi; Uchimura, Seiichi; Noguchi, Masahiro; Demura, Taku

    2016-01-01

    Microtubules assemble into several distinct arrays that play important roles in cell division and cell morphogenesis. To decipher the mechanisms that regulate the dynamics and organization of this versatile cytoskeletal component, it is essential to establish in vitro assays that use functional tubulin. Although plant tubulin has been purified previously from protoplasts by reversible taxol-induced polymerization, a simple and efficient purification method has yet to be developed. Here, we used a Tumor Overexpressed Gene (TOG) column, in which the tubulin-binding domains of a yeast (Saccharomyces cerevisiae) TOG homolog are immobilized on resin, to isolate functional plant tubulin. We found that several hundred micrograms of pure tubulin can readily be purified from cell suspension cultures of tobacco (Nicotiana tabacum) and Arabidopsis (Arabidopsis thaliana). The tubulin purified by the TOG column showed high assembly competence, partly because of low levels of polymerization-inhibitory phosphorylation of α-tubulin. Compared with porcine brain tubulin, Arabidopsis tubulin is highly dynamic in vitro at both the plus and minus ends, exhibiting faster shrinkage rates and more frequent catastrophe events, and exhibits frequent spontaneous nucleation. Furthermore, our study shows that an internal histidine tag in α-tubulin can be used to prepare particular isotypes and specifically engineered versions of α-tubulin. In contrast to previous studies of plant tubulin, our mass spectrometry and immunoblot analyses failed to detect posttranslational modification of the isolated Arabidopsis tubulin or detected only low levels of posttranslational modification. This novel technology can be used to prepare assembly-competent, highly dynamic pure tubulin from plant cell cultures. PMID:26747285

  18. Up-scaling single cell-inoculated suspension culture of human embryonic stem cells.

    Science.gov (United States)

    Singh, Harmeet; Mok, Pamela; Balakrishnan, Thavamalar; Rahmat, Siti Norfiza Binte; Zweigerdt, Robert

    2010-05-01

    We have systematically developed single cell-inoculated suspension cultures of human embryonic stem cells (hESC) in defined media. Cell survival was dependent on hESC re-aggregation. In the presence of the Rho kinase inhibitor Y-27632 (Ri) only approximately 44% of the seeded cells were rescued, but an optimized heat shock treatment combined with Ri significantly increased cell survival to approximately 60%. Mechanistically, our data suggest that E-cadherin plays a role in hESC aggregation and that dissociation and re-aggregation upon passaging functions as a purification step towards a pluripotency markers-enriched population. Mass expansion of hESC was readily achieved by up-scaling 2 ml cultures to serial passaging in 50 ml spinner flasks. A media comparison revealed that mTeSR was superior to KnockOut-SR in supporting cell proliferation and pluripotency. Persistent expression of pluripotency markers was achieved for two lines (hES2, hES3) that were used at higher passages (>86). In contrast, rapid down regulation of Oct4, Tra-1-60, and SSEA4 was observed for ESI049, a clinically compliant line, used at passages 20-36. The up-scaling strategy has significant potential to provide pluripotent cells on a clinical scale. Nevertheless, our data also highlights a significant line-to-line variability and the need for a critical assessment of novel methods with numerous relevant cell lines. Copyright 2010 Elsevier B.V. All rights reserved.

  19. Proteins differentially expressed in elicited cell suspension culture of Podophyllum hexandrum with enhanced podophyllotoxin content

    Directory of Open Access Journals (Sweden)

    Bhattacharyya Dipto

    2012-05-01

    Full Text Available Abstract Background Podophyllotoxin (PTOX, the precursor for semi-synthesis of cancer therapeutics like etoposide, teniposide and etophos, is primarily obtained from an endangered medicinal herb, Podophyllum hexandrum Royle. PTOX, a lignan is biosynthetically derived from the phenylpropanoid pathway. The aim of this study is to investigate changes in the P. hexandrum cell proteome potentially related to PTOX accumulation in response to methyl jasmonate (MeJA elicitation. High-resolution two-dimensional gel electrophoresis (2-DE followed by colloidal Coomassie staining and mass spectrometric analysis was used to detect statistically significant changes in cell’s proteome. Result The HPLC analysis showed approximately 7–8 fold change in accumulation of PTOX, in the 12day old cell suspension culture (i.e. after 9days of elicitation elicited with 100 μM MeJA as compared to the control. Using 2-DE a total of 233 spots was detected, out of which 105 spots were identified by MALDI TOF-TOF MS/MS. Data were subjected to functional annotation from a biological point of view through KEGG. The phenylpropanoid and monolignol pathway enzymes were identified, amongst these, chalcone synthase, polyphenol oxidase, caffeoyl CoA 3-O-methyltransferase, S-adenosyl-L-methionine-dependent methyltransferases, caffeic acid-O-methyl transferase etc. are noted as important. The relation of other differentially accumulated proteins with varied effects caused by elicitors on P. hexandrum cells namely stress and defense related protein, transcription and DNA replication and signaling are also discussed. Conclusions Elicitor-induced PTOX accumulation in P. hexandrum cell cultures provides a responsive model system to profile modulations in proteins related to phenylpropanoid/monolignol biosynthesis and other defense responses. Present findings form a baseline for future investigation on a non-sequenced medicinal herb P. hexandrum at molecular level.

  20. Critical factors besides treatment dose and duration need to be controlled in Pb toxicity tests in plant cell suspension cultures

    Directory of Open Access Journals (Sweden)

    Klaudia Sychta

    2017-08-01

    When effective illumination was adjusted to correct for turbidity at the highest lead concentration and pH was adjusted to 5.7–5.8, cell viability decreased with the increase of Pb(NO32 concentration and with treatment time. These experiments demonstrate that the toxic action of lead on cells in suspension depends strongly on culture conditions, and not only on the metal concentration and duration of treatment.

  1. Effects of aluminum on growth, polyamine metabolism, and inorganic ions in suspension cultures of red spruce (Picea rubens)

    Science.gov (United States)

    Rakesh Minocha; Walter C. Shortle; Daniel J. Jr. Coughin; Subhash C. Minocha

    1996-01-01

    The influence of age of red spruce (Picea rubens Sarg.) cell suspensions on aluminum (Al) effects was studied by adding AICI3 (0.2, 0.5, and 1.0 mM) to the media on each day of a 7-day culture period and analyzing for changes in total cell mass, polyamines, arginine decarboxylase activity, and inorganic ions after 24 h of...

  2. Plasma Membrane Proteomics of Arabidopsis Suspension-Cultured Cells Associated with Growth Phase Using Nano-LC-MS/MS.

    Science.gov (United States)

    Li, Bin; Takahashi, Daisuke; Kawamura, Yukio; Uemura, Matsuo

    2018-01-01

    Arabidopsis thaliana suspension-cultured cells (T87 line) are important model system for studies of responses to biotic and abiotic stresses at the cellular level in vitro since the cells have certain advantages compared with the whole plant system. However, the physiological and morphological characteristics of the cells are influenced by the progress of the growth phase of cells, which may result in different stress tolerance. To obtain comprehensive proteome profiles of the plasma membrane of Arabidopsis thaliana T87 suspension-cultured cells at the lag, log, or stationary growth phase, a shotgun proteomics method using nano-LC-MS/MS is used. The results obtained indicate that proteome profiles of the plasma membrane with the progress of the growth phase of cells dynamically changed, which may be associated with the physiological and morphological characteristics of the plasma membrane of the suspension-cultured cells. The proteomics results are further applied to explain different responsive patterns in the plasma membrane to cold acclimation and ABA treatment, which lead to understanding of different freezing tolerance associated with the growth phase of the cells.

  3. Wireless Remote Monitoring System for Cultural Heritage

    Directory of Open Access Journals (Sweden)

    Allan HUYNH

    2010-07-01

    Full Text Available Existing systems to collect temperature and relative humidity data at cultural heritage buildings require technical knowledge by people who are working with it, which is very seldom that they do have. The systems available today also require manual downloading of the collected data from the sensor to a computer for central storage and for further analysis. In this paper a wireless remote sensor network based on the ZigBee technology together with a simplified data collection system is presented. The system does not require any knowledge by the building administrator after the network is deployed. The wireless sensor device will automatically join available network when the user wants to expand the network. The collected data will be automatically and periodically synchronized to a remote main server via an Internet connection. The data can be used for centralized monitoring and other purpose. The power consumption of the sensor module is also minimized and the battery lifetime is estimated up to 10 years.

  4. Embryogenic callus formation, growth and regeneration in callus and suspension cultures of Miscanthus x ogiformis Honda 'Giganteus' as affected by proline

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted; Krogstrup, Peter; Hansen, Jürgen

    1997-01-01

    to test the effect of proline in suspension cultures. The proline additions affected the formation of embryogenic callus and the growth of suspension cultures. Improvements depended on the proline concentration and the basal salts of the medium. Addition of 12.5 to 50 mM proline to callus induction medium...... with Murashige and Skoog salts increased embryogenic callus formation on shoot apices and leaf explants while proline had no effect on embryogenic callus formation in medium with N6 salts. Increased growth with increasing proline concentration was obtained in suspension aggregates grown in medium with N6 salts......The effects of proline additions to culture systems of Miscanthus x ogiformis Honda Giganteus' were investigated. Proline was added in concentrations of 0, 12.5, 25, 50, 100 or 300 mM to the callus induction and suspension culture media containing either Murashige and Skoog or N6 basal salts and 22...

  5. Enhanced biosynthesis of withanolides by elicitation and precursor feeding in cell suspension culture of Withania somnifera (L. Dunal in shake-flask culture and bioreactor.

    Directory of Open Access Journals (Sweden)

    Ganeshan Sivanandhan

    Full Text Available The present study investigated the biosynthesis of major and minor withanolides of Withania somnifera in cell suspension culture using shake-flask culture and bioreactor by exploiting elicitation and precursor feeding strategies. Elicitors like cadmium chloride, aluminium chloride and chitosan, precursors such as cholesterol, mevalonic acid and squalene were examined. Maximum total withanolides detected [withanolide A (7606.75 mg, withanolide B (4826.05 mg, withaferin A (3732.81 mg, withanone (6538.65 mg, 12 deoxy withanstramonolide (3176.63 mg, withanoside IV (2623.21 mg and withanoside V (2861.18 mg] were achieved in the combined treatment of chitosan (100 mg/l and squalene (6 mM along with 1 mg/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose in culture at 4 h and 48 h exposure times respectively on 28th day of culture in bioreactor. We obtained higher concentrations of total withanolides in shake-flask culture (2.13-fold as well as bioreactor (1.66-fold when compared to control treatments. This optimized protocol can be utilized for commercial level production of withanolides from suspension culture using industrial bioreactors in a short culture period.

  6. Enhanced Biosynthesis of Withanolides by Elicitation and Precursor Feeding in Cell Suspension Culture of Withania somnifera (L.) Dunal in Shake-Flask Culture and Bioreactor

    Science.gov (United States)

    Sivanandhan, Ganeshan; Selvaraj, Natesan; Ganapathi, Andy; Manickavasagam, Markandan

    2014-01-01

    The present study investigated the biosynthesis of major and minor withanolides of Withania somnifera in cell suspension culture using shake-flask culture and bioreactor by exploiting elicitation and precursor feeding strategies. Elicitors like cadmium chloride, aluminium chloride and chitosan, precursors such as cholesterol, mevalonic acid and squalene were examined. Maximum total withanolides detected [withanolide A (7606.75 mg), withanolide B (4826.05 mg), withaferin A (3732.81 mg), withanone (6538.65 mg), 12 deoxy withanstramonolide (3176.63 mg), withanoside IV (2623.21 mg) and withanoside V (2861.18 mg)] were achieved in the combined treatment of chitosan (100 mg/l) and squalene (6 mM) along with 1 mg/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose in culture at 4 h and 48 h exposure times respectively on 28th day of culture in bioreactor. We obtained higher concentrations of total withanolides in shake-flask culture (2.13-fold) as well as bioreactor (1.66-fold) when compared to control treatments. This optimized protocol can be utilized for commercial level production of withanolides from suspension culture using industrial bioreactors in a short culture period. PMID:25089711

  7. Enhanced Production of Bioactive Isoprenoid Compounds from Cell Suspension Cultures of Artemisia annua L. Using β-Cyclodextrins

    Science.gov (United States)

    Rizzello, Francesca; De Paolis, Angelo; Durante, Miriana; Blando, Federica; Mita, Giovanni; Caretto, Sofia

    2014-01-01

    Plant cell cultures as valuable tools for the production of specific metabolites can be greatly improved by the application of elicitors including cyclodextrins (CDs) for enhancing the yields of the desired plant compounds. Here the effects of 2,6-dimethyl-β-cyclodextrins (DIMEB) on the production of carotenoids and quinones from Artemisia annua L. cell suspension cultures were investigated. The addition of 50 mM DIMEB induced an early increase of intracellular carotenoid and quinone contents, which could be observed to a higher extent for lutein (10-fold), Q9 (3-fold) and Q10 (2.5-fold). Real Time PCR analysis revealed that the expression of 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) gene in DIMEB treated cell cultures after three days was 2.5-fold higher than in untreated samples, thus suggesting that the DIMEB induced increase of carotenoids and quinones could be due to the induction of the plastidial isoprenoid biosynthetic route. In addition, the DIMEB treatment induced an enhanced release of carotenoids and quinones into the culture medium of A. annua cell suspension cultures possibly due to the ability of CDs to form inclusion complexes with hydrophobic molecules. PMID:25338048

  8. Effect of Different Carbon Sources on Relative Growth Rate, Internal Carbohydrates, and Mannitol 1-Oxidoreductase Activity in Celery Suspension Cultures.

    Science.gov (United States)

    Stoop, JMH.; Pharr, D. M.

    1993-11-01

    Little information exists concerning the biochemical route of mannitol catabolism in higher plant cells. In this study, the role of a recently discovered mannitol 1-oxidoreductase (MDH) in mannitol catabolism was investigated. Suspension cultures of celery (Apium graveolens L. var dulce [Mill.] Pers.) were successfully grown on nutrient media with either mannitol, mannose, or sucrose as the sole carbon source. Cell cultures grown on any of the three carbon sources did not differ in relative growth rate, as measured by packed cell volume, but differed drastically in internal carbohydrate concentration. Mannitol-grown cells contained high concentrations of mannitol and extremely low concentrations of sucrose, fructose, glucose, and mannose. Sucrose-grown cells had high concentrations of sucrose early in the growth cycle and contained a substantial hexose pool. Mannose-grown cells had a high mannose concentration early in the cycle, which decreased during the growth cycle, whereas their internal sucrose concentrations remained relatively constant during the entire growth cycle. Celery suspension cultures on all three carbon substrates contained an NAD-dependent MDH. Throughout the growth cycle, MDH activity was 2- to 4-fold higher in mannitol-grown cells compared with sucrose- or mannose-grown cells, which did not contain detectable levels of mannitol, indicating that MDH functions pre-dominantly in an oxidative capacity in situ. The MDH activity observed in celery cells was 3-fold higher than the minimum amount required to account for the observed rate of mannitol utilization from the media. Cultures transferred from mannitol to mannose underwent a decrease in MDH activity over a period of days, and transfer from mannose to mannitol resulted in an increase in MDH activity. These data provide strong evidence that MDH plays an important role in mannitol utilization in celery suspension cultures.

  9. Glycyrrhiza glabra (Linn.) and Lavandula officinalis (L.) cell suspension cultures-based biotransformation of β-artemether.

    Science.gov (United States)

    Patel, Suman; Gaur, Rashmi; Upadhyaya, Mohita; Mathur, Archana; Mathur, Ajay K; Bhakuni, Rajendra S

    2011-07-01

    The biotransformation of β-artemether (1) by cell suspension cultures of Glycyrrhiza glabra and Lavandula officinalis is reported here for the first time. The major biotransformed product appeared as a grayish-blue color spot on thin-layer chromatography (TLC) with transparent crystal-like texture. Based on its infrared (IR) and (1)H nuclear magnetic resonance (NMR) spectra, the product was characterized as a tetrahydrofuran (THF)-acetate derivative (2). The highest conversion efficiencies of 57 and 60% were obtained when 8-9-day-old cell suspensions of G. glabra and L. officinalis were respectively fed with 4-7 mg of compound 1 in 40 ml of medium per culture and the cells were harvested after 2-5 days of incubation. The addition of compound 1 at the beginning of the culture cycle caused severe growth depression in a dose-dependent manner, resulting in poor bioconversion efficiency of ~25% at 2-5 mg/culture dose only.

  10. Signal transduction in artichoke [Cynara cardunculus L. subsp. scolymus (L.) Hayek] callus and cell suspension cultures under nutritional stress.

    Science.gov (United States)

    Lattanzio, Vincenzo; Caretto, Sofia; Linsalata, Vito; Colella, Giovanni; Mita, Giovanni

    2018-03-16

    Stimulated production of secondary phenolic metabolites and proline was studied by using cell cultures of artichoke [Cynara cardunculus L. subsp. scolymus (L.) Hayek] submitted to nutritional stress. Artichoke cell cultures accumulated phenolic secondary metabolites in a pattern similar to that seen in artichoke leaves and heads (capitula). This paper shows that both callus and cell suspension cultures under nutritional stress accumulated phenolic compounds and proline, at the same time their biomass production was negatively affected by nutrient deficiency. The results obtained strongly suggest that plant tissues respond to nutrient deprivation by a defensive costly mechanism, which determines the establishment of a mechanism of trade-off between growth and adaptive response. Furthermore, the results of this research suggest that perception of abiotic stress and increased phenolic metabolites are linked by a sequence of biochemical processes that also involves the intracellular free proline and the oxidative pentose phosphate pathway. The main conclusion of this paper is that, once calli and cell suspension cultures respond to nutrient deficiency, in acclimated cells the establishment of a negative correlation between primary metabolism (growth) and secondary metabolism (defence compounds) is observed. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  11. Idaho National Laboratory Cultural Resource Monitoring Report for FY 2009

    Energy Technology Data Exchange (ETDEWEB)

    Brenda R. Pace; Julie B. Braun

    2009-10-01

    This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during fiscal year 2009 (FY 2009). Throughout the year, thirty-eight cultural resource localities were revisited including: two locations with Native American human remains, one of which is a cave, two additional caves, twenty-two prehistoric archaeological sites, six historic homesteads, two historic stage stations, two historic trails, and two nuclear resources, including Experimental Breeder Reactor-I, which is a designated National Historic Landmark. Several INL project areas were also monitored in FY 2009 to assess project compliance with cultural resource recommendations and monitor the effects of ongoing project activities. Although impacts were documented at a few locations and trespassing citations were issued in one instance, no significant adverse effects that would threaten the National Register eligibility of any resources were observed. Monitoring also demonstrated that several INL projects generally remain in compliance with recommendations to protect cultural resources.

  12. Induction and Analysis of the Alkaloid Mitragynine Content of a Mitragyna speciosa Suspension Culture System upon Elicitation and Precursor Feeding

    Directory of Open Access Journals (Sweden)

    Nor Nahazima Mohamad Zuldin

    2013-01-01

    Full Text Available This study aimed to determine the effects of different concentrations and combinations of the phytohormones 2,4-dichlorophenoxy acetic acid (2,4-D, kinetin, 6-benzylaminopurine (BAP, and 1-naphthaleneacetic acid (NAA on callus induction and to demonstrate the role of elicitors and exogenous precursors on the production of mitragynine in a Mitragyna speciosa suspension culture. The best callus induction was achieved from petiole explants cultured on WPM that was supplemented with 4 mg L−1 2, 4-D (70.83%. Calli were transferred to liquid media and agitated on rotary shakers to establish Mitragyna speciosa cell suspension cultures. The optimum settled cell volume was achieved in the presence of WPM that contained 3 mg L−1 2,4-D and 3% sucrose (9.47±0.4667 mL. The treatment of cultures with different concentrations of yeast extract and salicylic acid for different inoculation periods revealed that the highest mitragynine content as determined by HPLC was achieved from the culture treated with 250 mg L−1 yeast extract (9.275±0.082 mg L−1 that was harvested on day 6 of culturing; salicylic acid showed low mitragynine content in all concentrations used. Tryptophan and loganin were used as exogenous precursors; the highest level of mitragynine production was achieved in cultures treated with 3 μM tryptophan and harvested at 6 days (13.226±1.98 mg L−1.

  13. Propagation of Brazilian Zika virus strains in static and suspension cultures using Vero and BHK cells.

    Science.gov (United States)

    Nikolay, Alexander; Castilho, Leda R; Reichl, Udo; Genzel, Yvonne

    2017-03-23

    The recent spread of Zika virus (ZIKV) in the Americas and the Pacific has reached alarming levels in more than 60 countries. However, relatively little is known about the disease on a virological and epidemiological level and its consequences for humans. Accordingly, a large demand for in vitro derived Brazilian ZIKV material to support in vitro and in vivo studies has arisen. However, a prompt supply of ZIKV and ZIKV antigens cannot be guaranteed as the production of this virus typically using Vero or C6/36 cell lines remains challenging. Here we present a production platform based on BHK-21 suspension (BHK-21 SUS ) cells to propagate Brazilian ZIKV at larger quantities in perfusion bioreactors. Scouting experiments performed in tissue culture flasks using adherent BHK-21 and Vero cells have demonstrated similar permissivity and virus yields for four different Brazilian ZIKV isolates. The cell-specific yield of infectious virus particles varied between respective virus strains (1-48PFU/cell), and the ZIKV isolate from the Brazilian state Pernambuco (ZIKV PE ) showed to be a best performing isolate for both cell lines. However, infection studies of BHK-21 SUS cells with ZIKV PE in shake flasks resulted in poor virus replication, with a maximum titer of 8.9×10 3 PFU/mL. Additional RT-qPCR measurements of intracellular and extracellular viral RNA levels revealed high viral copy numbers within the cell, but poor virus release. Subsequent cultivation in a perfusion bioreactor using an alternating tangential flow filtration system (ATF) under controlled process conditions enabled cell concentrations of about 1.2×10 7 cells/mL, and virus titers of 3.9×10 7 PFU/mL. However, while the total number of infectious virus particles was increased, the cell-specific yield (3.3PFU/cell) remained lower than determined in adherent cell lines. Nevertheless, the established perfusion process allows to provide large amounts of ZIKV material for research and is a first step towards

  14. Idaho National Laboratory Cultural Resource Monitoring Report for 2013

    Energy Technology Data Exchange (ETDEWEB)

    Williams, Julie B. [Idaho National Lab. (INL), Idaho Falls, ID (United States)

    2013-10-01

    This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during 2013. Throughout the year, thirty-eight cultural resource localities were revisited including: two locations with Native American human remains, one of which is also a cave; fourteen additional caves; seven prehistoric archaeological sites ; four historic archaeological sites; one historic trail; one nuclear resource (Experimental Breeder Reactor-I, a designated National Historic Landmark); and nine historic structures located at the Central Facilities Area. Of the monitored resources, thirty-three were routinely monitored, and five were monitored to assess project compliance with cultural resource recommendations along with the effects of ongoing project activities. On six occasions, ground disturbing activities within the boundaries of the Power Burst Facility/Critical Infrastructure Test Range Complex (PBF/CITRC) were observed by INL CRM staff prepared to respond to any additional finds of Native American human remains. In addition, two resources were visited more than once as part of the routine monitoring schedule or to monitor for additional damage. Throughout the year, most of the cultural resources monitored had no visual adverse changes resulting in Type 1determinations. However, Type 2 impacts were noted at eight sites, indicating that although impacts were noted or that a project was operating outside of culturally cleared limitations, cultural resources retained integrity and noted impacts did not threaten National Register eligibility. No new Type 3 or any Type 4 impacts that adversely impacted cultural resources and threatened National Register eligibility were observed at cultural resources monitored in 2013.

  15. Biotransformation of artemisinin using cell suspension cultures of Catharanthus roseus (L.) G.Don and Lavandula officinalis L.

    Science.gov (United States)

    Patel, Suman; Gaur, Rashmi; Verma, Priyanka; Bhakuni, Rajendra S; Mathur, Archana

    2010-08-01

    Artemisinin, an antimalarial compound, at 5 mg/40 ml, was transformed by cell suspension cultures of Catharanthus roseus (L.) G.Don and Lavandula officinalis L. into deoxyartemisinin with yields >78% (3.93 mg deoxyartemisinin from 5 mg artemisinin). Maximum conversion (78.6 and 78%) occurred after 6 and 7 days of adding artemisinin to 20 and 9 days old cultures of C. roseus and L. officinalis, respectively. The procedure was scaled up by and 500 mg artemisinin was transformed into 390 mg deoxyartemisinin. Addition of artemisinin at the beginning of the culture cycle resulted in >50% reduction in dry biomass production with no bioconversion. Conversion of artemisinin occurred intracellularly followed by leaching of the product into the medium.

  16. Efficient embryogenic suspension culturing and rapid transformation of a range of elite genotypes of sweet potato (Ipomoea batatas [L.] Lam.).

    Science.gov (United States)

    Yang, Jun; Bi, Hui-Ping; Fan, Wei-Juan; Zhang, Min; Wang, Hong-Xia; Zhang, Peng

    2011-12-01

    Efficient Agrobacterium tumefaciens-mediated transformation was developed using embryogenic suspension cell cultures of elite sweet potato (Ipomoea batatas [L.] Lam.) cultivars, including Ayamurasaki, Sushu2, Sushu9, Sushu11, Wanshu1, Xushu18 and Xushu22. Embryogenic suspension cultures were established in LCP medium using embryogenic calli induced from apical or axillary buds on an induction medium containing 2 mg l(-1) 2,4-D. Suspension cultures were co-cultivated with A. tumefaciens strain LBA4404 harboring the binary plasmid pCAMBIA1301 with the hpt gene as a selectable marker and an intron-interrupted uidA gene as a visible marker. Several key steps of the sweet potato transformation system have been investigated and optimized, including the appropriate antibiotics and their concentrations for suppressing Agrobacterium growth and the optimal doses of hygromycin for transformant selection. A total of 485 putative transgenic plant lines were produced from the transformed calli via somatic embryogenesis and germination to plants under 10 mg l(-1) hygromycin and 200 mg l(-1) cefotaxime. PCR, GUS and Southern blot analyses of the regenerated plants showed that 92.35% of them were transgenic. The number of T-DNA insertions varied from one to three in most transgenic plant lines. Plants showed 100% survival when 308 transgenics were transferred to soil in the greenhouse and then to the field. Most of them were morphologically normal, with the production of storage roots after 3 months of cultivation in the greenhouse or fields. The development of such a robust transformation method suitable to a range of sweet potato genotypes not only provides a routine tool for genetic improvement via transgenesis but also allows us to conduct a functional verification of endogenous genes in sweet potato. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  17. EPR monitoring of the bioavailability of an organic xenobiotic (4-hydroxy-TEMPO) in model clay suspensions and pastes

    International Nuclear Information System (INIS)

    Dumestre, Alain; Spagnuolo, Matteo; Bladon, Rebecca; Berthelin, Jacques; Baveye, Philippe

    2006-01-01

    Electron paramagnetic resonance spectroscopy is used to monitor the bioavailability of a nitroxide spin probe, 4-hydroxy-Tempo or Tempol, in Ca-hectorite suspensions and pastes, to bacteria capable of degrading this probe co-metabolically. In nutrient solutions with an initial probe concentration of 1.2 mM and in the absence of hectorite, bacteria are able to denature Tempol and eliminate its paramagnetic signal within 48 h. In the presence of hectorite and after flocculation, the effect of bacteria is significantly delayed, but almost complete denaturation still occurs, after roughly 120 h. When hectorite is added but the bacterial/clay suspension is not centrifuged, Tempol denaturation levels off after about 24 h and reaches a plateau with approximately 45% of Tempol remaining. This plateau does not constitute evidence of limited bioavailability, as is widely assumed, since subsequent addition of nutrients causes the denaturation reaction to proceed to a second plateau, with merely 10% of Tempol remaining. - Spectroscopic data demonstrate that the bioavailability of an organic compound in clay suspensions and pastes strongly depends on conditions that affect bacterial metabolism

  18. EPR monitoring of the bioavailability of an organic xenobiotic (4-hydroxy-TEMPO) in model clay suspensions and pastes

    Energy Technology Data Exchange (ETDEWEB)

    Dumestre, Alain [Laboratory of Geoenvironmental Science and Engineering, Bradfield Hall, Cornell University, Ithaca, NY 14853 (United States); Spagnuolo, Matteo [Dipartimento di Biologia e Chimica Agro-forestale ed Ambientale, Universita degli Studi di Bari, via Amendola 165/a, 70126 Bari (Italy); Bladon, Rebecca [158 Kottinger Drive, Pleasanton, CA (United States); Berthelin, Jacques [CNRS-LIMOS Laboratoire des Interactions Microorganismes-Mineraux-Matieres Organiques, UMR 7137 du CNRS, Universite Henri Poincare, B.P. 239, F-54506 Vandoeuvre les Nancy (France); Baveye, Philippe [Laboratory of Geoenvironmental Science and Engineering, Bradfield Hall, Cornell University, Ithaca, NY 14853 (United States)]. E-mail: philippe.baveye@cornell.edu

    2006-09-15

    Electron paramagnetic resonance spectroscopy is used to monitor the bioavailability of a nitroxide spin probe, 4-hydroxy-Tempo or Tempol, in Ca-hectorite suspensions and pastes, to bacteria capable of degrading this probe co-metabolically. In nutrient solutions with an initial probe concentration of 1.2 mM and in the absence of hectorite, bacteria are able to denature Tempol and eliminate its paramagnetic signal within 48 h. In the presence of hectorite and after flocculation, the effect of bacteria is significantly delayed, but almost complete denaturation still occurs, after roughly 120 h. When hectorite is added but the bacterial/clay suspension is not centrifuged, Tempol denaturation levels off after about 24 h and reaches a plateau with approximately 45% of Tempol remaining. This plateau does not constitute evidence of limited bioavailability, as is widely assumed, since subsequent addition of nutrients causes the denaturation reaction to proceed to a second plateau, with merely 10% of Tempol remaining. - Spectroscopic data demonstrate that the bioavailability of an organic compound in clay suspensions and pastes strongly depends on conditions that affect bacterial metabolism.

  19. Critical factors besides treatment dose and duration need to be controlled in Pb toxicity tests in plant cell suspension cultures

    OpenAIRE

    Klaudia Sychta; Janusz Szklarzewicz; Aneta Słomka; Ewa Gregoraszczuk; Elżbieta Kuta

    2017-01-01

    The study was designed to determine the proper conditions for suspension culture of Viola tricolor cells in toxicity studies of Pb at different concentrations (0, 200, 500, 1000, 2000 µM) and exposure times (24, 48, 72 h). By forming insoluble salts with ions from the medium, lead (II) nitrate added to the medium decreased the initial 5.7–5.8 pH of the medium, depending on the Pb salt concentration and light intensity. In alamarBlue assays, we found no dose- or time-dependent effect of Pb...

  20. Experimental infection of New Zealand Merino sheep with a suspension of Mycobacterium avium subspecies paratuberculosis (Map) strain Telford: Kinetics of the immune response, histopathology and Map culture.

    Science.gov (United States)

    Dukkipati, Venkata S R; Ridler, Anne L; Thompson, Keith G; Buddle, Bryce M; Hedgespeth, Barry A; Price-Carter, Marian; Begg, Douglas J; Whittington, Richard J; Gicquel, Brigitte; Murray, Alan

    2016-11-15

    A long-term study was undertaken to monitor immune responses, faecal cultures and clinical disease in sheep experimentally infected with Mycobacterium avium subspecies paratuberculosis (Map) strain Telford. New Zealand Merino lambs (N=56) were challenged with three oral doses of Map suspension. The lambs were weighed and faecal and blood samples obtained at different time-points. At 63 weeks post-challenge, surviving sheep were euthanised and samples of liver, ileo-caecal valve and mesenteric lymph node were collected for histopathology and Map culture. High IFN-γ and antibody responses were evident as early as 8 weeks post-C1 which persisted until the end of the trial. Approximately 92% of the sheep shed Map in faeces at 36 weeks post-challenge, with the prevalence decreasing to around 40% at the end of the trial. Thirteen sheep progressively lost weight and were euthanised between weeks 32 and 58 post-challenge. Nearly 58% of surviving sheep exhibited histo-pathological lesions in at least one of the three tissues sampled, while 42% showed acid-fast bacilli in at least one tissue. A positive Map culture in at least one tissue was obtained from approximately 85% of sheep. These results indicate that the three doses of Map challenge were highly effective in establishing Johne's disease in NZ Merino lambs. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  1. Altered nitrogen metabolism associated with de-differentiated suspension cultures derived from root cultures of Datura stramonium studied by heteronuclear multiple bond coherence (HMBC) NMR spectroscopy.

    Science.gov (United States)

    Fliniaux, Ophélie; Mesnard, François; Raynaud-Le Grandic, Sophie; Baltora-Rosset, Sylvie; Bienaimé, Christophe; Robins, Richard J; Fliniaux, Marc-André

    2004-05-01

    De-differentiation of transformed root cultures of Datura stramonium has previously been shown to cause a loss of tropane alkaloid synthetic capacity. This indicates a marked shift in physiological status, notably in the flux of primary metabolites into tropane alkaloids. Nitrogen metabolism in transformed root cultures of D. stramonium (an alkaloid-producing system) and de-differentiated suspension cultures derived therefrom (a non-producing system) has been compared using Nuclear Magnetic Resonance (NMR) spectroscopy. (15)N-Labelled precursors [((15)NH(4))(2)SO(4) and K(15)NO(3)] were fed and their incorporation into nitrogenous metabolites studied using Heteronuclear Multiple Bond Coherence (HMBC) NMR spectroscopy. In both cultures, the same amino acids were resolved in the HMBC spectra. However, marked differences were found in the intensity of labelling of a range of nitrogenous compounds. In differentiated root cultures, cross-peaks corresponding to secondary metabolites, such as tropine, were observed, whereas these were absent in the de-differentiated cultures. By contrast, N- acetylputrescine and gamma-aminobutyric acid (GABA) accumulated in the de-differentiated cultures to a much larger extent than in the root cultures. It can therefore be suggested that the loss of alkaloid biosynthesis was compensated by the diversion of putrescine metabolism away from the tropane pathway and toward the synthesis of GABA via N-acetylputrescine.

  2. Monitoring of cell cultures with LTCC microelectrode array.

    Science.gov (United States)

    Ciosek, P; Zawadzki, K; Łopacińska, J; Skolimowski, M; Bembnowicz, P; Golonka, L J; Brzózka, Z; Wróblewski, W

    2009-04-01

    Monitoring of cell cultures in microbioreactors is a crucial task in cell bioassays and toxicological tests. In this work a novel tool based on a miniaturized sensor array fabricated using low-temperature cofired ceramics (LTCC) technology is presented. The developed device is applied to the monitoring of cell-culture media change, detection of the growth of various species, and in toxicological studies performed with the use of cells. Noninvasive monitoring performed with the LTCC microelectrode array can be applied for future cell-engineering purposes.

  3. Rotary orbital suspension culture of embryonic stem cell-derived neural stem/progenitor cells: impact of hydrodynamic culture on aggregate yield, morphology and cell phenotype.

    Science.gov (United States)

    Laundos, Tiago L; Silva, Joana; Assunção, Marisa; Quelhas, Pedro; Monteiro, Cátia; Oliveira, Carla; Oliveira, Maria J; Pêgo, Ana P; Amaral, Isabel F

    2017-08-01

    Embryonic stem (ES)-derived neural stem/progenitor cells (ES-NSPCs) constitute a promising cell source for application in cell therapies for the treatment of central nervous system disorders. In this study, a rotary orbital hydrodynamic culture system was applied to single-cell suspensions of ES-NSPCs, to obtain homogeneously-sized ES-NSPC cellular aggregates (neurospheres). Hydrodynamic culture allowed the formation of ES-NSPC neurospheres with a narrower size distribution than statically cultured neurospheres, increasing orbital speeds leading to smaller-sized neurospheres and higher neurosphere yield. Neurospheres formed under hydrodynamic conditions (72 h at 55 rpm) showed higher cell compaction and comparable percentages of viable, dead, apoptotic and proliferative cells. Further characterization of cellular aggregates provided new insights into the effect of hydrodynamic shear on ES-NSPC behaviour. Rotary neurospheres exhibited reduced protein levels of N-cadherin and β-catenin, and higher deposition of laminin (without impacting fibronectin deposition), matrix metalloproteinase-2 (MMP-2) activity and percentage of neuronal cells. In line with the increased MMP-2 activity levels found, hydrodynamically-cultured neurospheres showed higher outward migration on laminin. Moreover, when cultured in a 3D fibrin hydrogel, rotary neurospheres generated an increased percentage of neuronal cells. In conclusion, the application of a constant orbital speed to single-cell suspensions of ES-NSPCs, besides allowing the formation of homogeneously-sized neurospheres, promoted ES-NSPC differentiation and outward migration, possibly by influencing the expression of cell-cell adhesion molecules and the secretion of proteases/extracellular matrix proteins. These findings are important when establishing the culture conditions needed to obtain uniformly-sized ES-NSPC aggregates, either for use in regenerative therapies or in in vitro platforms for biomaterial development or

  4. Panax ginseng Adventitious Root Suspension Culture: Protocol for Biomass Production and Analysis of Ginsenosides by High Pressure Liquid Chromatography.

    Science.gov (United States)

    Murthy, Hosakatte Niranjana; Paek, Kee Yoeup

    2016-01-01

    Panax ginseng C.A. Meyer (Korean ginseng) is a popular herbal medicine. It has been used in Chinese and Oriental medicines since thousands of years. Ginseng products are generally used as a tonic and an adaptogen to resist the adverse influence of a wide range of physical, chemical and biological factors, and to restore homeostasis. Ginsenosides or ginseng saponins are the principal active ingredients of ginseng. Since ginseng cultivation process is very slow and needs specific environment for field cultivation, cell and tissue cultures are sought as alternatives for the production of ginseng biomass and bioactive compounds. In this chapter, we focus on methods of induction of adventitious roots from ginseng roots, establishment of adventitious root suspension cultures using bioreactors, procedures for processing of adventitious roots, and analysis of ginsenosides by high pressure liquid chromatography.

  5. Idaho National Laboratory Cultural Resource Monitoring Report for FY 2010

    Energy Technology Data Exchange (ETDEWEB)

    INL Cultural Resource Management Office

    2010-10-01

    This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during fiscal year 2010 (FY 2010). Throughout the year, thirty-three cultural resource localities were revisited, including somethat were visited more than once, including: two locations with Native American human remains, one of which is a cave, two additional caves, twenty-six prehistoric archaeological sites, two historic stage stations, and Experimental Breeder Reactor-I, which is a designated National Historic Landmark. The resources that were monitored included seventeen that are routinely visited and sixteen that are located in INL project areas. Although impacts were documented at a few locations and one trespassing incident (albeit sans formal charges) was discovered, no significant adverse effects that would threaten the National Register eligibility of any resources were observed. Monitoring also demonstrated that several INL projects generally remain in compliance with recommendations to protect cultural resources.

  6. Opposite extremes in ethylene/nitric oxide ratio induce cell death in suspension culture and root apices of tomato exposed to salt stress.

    Science.gov (United States)

    Poór, P; Borbély, P; Kovács, Judit; Papp, Anita; Szepesi, Ágnes; Takács, Z; Tari, Irma

    2014-12-01

    The plant hormone ethylene or the gaseous signalling molecule nitric oxide (NO) may enhance salt stress tolerance by maintaining ion homeostasis, first of all K+/Na+ ratio of tissues. Ethylene and NO accumulation increased in the root apices and suspension culture cells of tomato at sublethal salt stress caused by 100 mM NaCl, however, the induction phase of programmed cell death (PCD) was different at lethal salt concentration. The production of ethylene by root apices and the accumulation of NO in the cells of suspension culture did not increase during the initiation of PCD after 250 mM NaCl treatment. Moreover, cells in suspension culture accumulated higher amount of reactive oxygen species which, along with NO deficiency contributed to cell death induction. The absence of ethylene in the apical root segments and the absence of NO accumulation in the cell suspension resulted in similar ion disequilibrium, namely K+/Na+ ratio of 1.41 ± 0.1 and 1.68 ± 0.3 in intact plant tissues and suspension culture cells, respectively that was not tolerated by tomato.

  7. Elicitation of Jerusalem artichoke (Helianthus tuberosus L.) cell suspension culture for enhancement of inulin production and altered degree of polymerisation.

    Science.gov (United States)

    Ma, Chunquan; Zhou, Dong; Wang, Haitao; Han, Dongming; Wang, Yang; Yan, Xiufeng

    2017-01-01

    Plant cell suspension cultures have emerged as a potential source of secondary metabolites for food additives and pharmaceuticals. In this study inulin accumulation and its degree of polymerisation (DP) in the treated cells in the same medium were investigated after treatment with six types of elicitors. An in vitro cell suspension culture of Jerusalem artichoke (Helianthus tuberosus L.) was optimised by adding an extra nitrogen source. According to the growth kinetics, a maximum biomass of 5.48 g L -1 was obtained from the optimal cell suspension medium consisted of Murashige and Skoog basic medium (MS) + 1.0 mg L -1 α-naphthalene acetic acid (NAA) + 1.0 mg L -1 6-benzylaminopurine (6-BA) + 0.5 mg L -1 proline + 1.0 mg L -1 glutamine. Methyl jasmonate (MeJA, 250 µmol L -1 ) treatment for 15 days led to the highest levels of inulin (2955.27 ± 9.81 mg L -1 compared to control of 1217.46 ± 0.26 mg L -1 ). The elicited effect of five elicitors to the suspension cells of Jerusalem artichoke is as follows: AgNO 3 (Ag, 10 µmol L -1 ), salicylic acid (SA, 75 µmol L -1 ), chitosan (KJT, 40 mg L -1 ), Trichoderma viride (Tv, 90 mg L -1 ), yeast extract (YE, 0.25 mg L -1 ), and the corresponding content of inulin is increased by 2.05-, 1.93-, 1.76-, 1.44- and 1.18-fold compared to control, respectively. The obvious effect on the percentage of lower DP in inulin was observed in cells treated with 40 mg L -1 KJT, 0.25 mg L -1 YE and 10 µmol L -1 Ag. Among the six types of elicitors, the descending order of inulin content is MeJA > Ag > SA > KJT > Tv > YE. For the purpose inulin with lower DP and its application to prebiotic food, three elicitors, including KJT, YE and Ag, can be used for the elicitation. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  8. Enhanced production of vanillin flavour metabolites by precursor feeding in cell suspension cultures of Decalepis hamiltonii Wight & Arn., in shake flask culture.

    Science.gov (United States)

    Matam, Pradeep; Parvatam, Giridhar; Shetty, Nandini P

    2017-12-01

    The flavour rich tuberous roots of Decalepis hamiltonii are known for its edible and medicinal use and have become endangered due to commercial over-exploitation. Besides 2-Hydroxy-4-methoxy benzaldehyde (2H4MB), other flavour metabolites in tuberous roots include vanillin, 4-Methoxy Cinnamic acid derivatives, aromatic alcohols etc. So far, there are no reports on the pathway of 2H4MB biosynthesis nor there is an organized work on biotransformation using normal and cell suspension cultures for obtaining these metabolites using precursors. The main aim of the study is to develop a method for enhanced production of flavour attributing metabolites through ferulic acid (FA) feeding to the D. hamiltonii callus culture medium. Biomass of D. hamiltonii cell suspension cultures was maximum (200.38 ± 1.56 g/l) by 4th week. Maximum production of 2H4MB was recorded on 4th week (0.08 ± 0.01 mg/100 g dry weight) as quantified by HPLC. Addition of 0.1-1.5 mM ferulic acid as precursor in the culture medium showed significant ( p  vanillin, 2H4MB, vanillic acid, ferulic acid were of 0.1 ± 0.02 mg/100 g, 0.44 ± 0.01 mg/100 g, 0.52 ± 0.04 mg/100 g, 0.18 ± 0.02 mg/100 g DW respectively in 4 weeks of cultured cells supplemented with 1 mM ferulic acid as a precursor. The results indicate that, substantial increase in the levels of flavour metabolites in D. hamiltonii callus suspension culture was achieved. This would be having implications in biosynthesis of respective vanilla flavour attributing metabolites at very high levels for their large scale production.

  9. Characterization of an immunomodulatory Der p 2-FIP-fve fusion protein produced in transformed rice suspension cell culture.

    Science.gov (United States)

    Su, Chin-Fen; Kuo, I-Chun; Chen, Peng-Wen; Huang, Chiung-Hui; Seow, See Voon; Chua, Kaw Yan; Yu, Su-May

    2012-02-01

    Der p 2, a major allergen of Dermatophagoides pteronyssinus mites, is one of the most clinically relevant allergens to allergic patients worldwide. FIP-fve protein (Fve) from the golden needle mushroom (Flammulina velutipes) is an immunomodulatory protein with potential Th1-skewed adjuvant properties. Here, we produced and immunologically evaluated a Der p 2-Fve fusion protein as a potential immunotherapeutic for allergic diseases. Using an inducible expression system in cultured rice suspension cells, the recombinant Der p 2-Fve fusion protein (designated as OsDp2Fve) was expressed in rice cells under the control of an α-amylase gene (αAmy8) promoter and secreted under sucrose starvation. OsDp2Fve was partially purified from the cultured medium. The conformation of Der p 2 in OsDp2Fve remains intact as reflected by its unaltered allergenicity, as assessed by human IgE ELISA and histamine release assays, compared to non-fusion Der p 2 protein. Furthermore, the Fve protein expressed in OsDp2Fve retains its in vitro lymphoproliferative activity but loses its hemagglutination and lymphoagglutination effects compared to the native protein. Notably, in vivo evaluation showed that mice administered with OsDp2Fve possessed an enhanced production of Der p 2-specific IgG antibodies without potentiating the production of Der p 2-specific IgE and Th2 effector cytokines in comparison with mice co-administered with native Fve and Der p 2 proteins. These results suggest that the recombinant Der p 2-Fve fusion protein produced in rice suspension cell cultures has a great potential for allergy immunotherapy.

  10. Idaho National Laboratory Cultural Resource Monitoring Report for FY 2008

    Energy Technology Data Exchange (ETDEWEB)

    Brenda R. Pace

    2009-01-01

    This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during fiscal year 2008 (FY 2008). Throughout the year, 45 cultural resource localities were revisited including: two locations of heightened Shoshone-Bannock tribal sensitivity, four caves, one butte, twenty-eight prehistoric archaeological sites, three historic homesteads, two historic stage stations, one historic canal construction camp, three historic trails, and Experimental Breeder Reactor-I, which is a designated National Historic Landmark. Several INL project areas were also monitored in FY 2008 to assess project compliance with cultural resource recommendations, confirm the locations of previously recorded cultural resources in relation to project activities, to assess the damage caused by fire-fighting efforts, and to watch for cultural materials during ground disturbing activities. Although impacts were documented at a few locations, no significant adverse effects that would threaten the National Register eligibility of any resource were observed. Monitoring also demonstrated that INL projects generally remain in compliance with recommendations to protect cultural resources

  11. Accumulation of ixerin F and activities of some terpenoid bisynthetic enzymes in a cell suspension culture of Lactuca virosa L.

    Directory of Open Access Journals (Sweden)

    Anna Stojakowska

    2014-01-01

    Full Text Available A cell suspension culture of Lactuca virosa L. (Asteraceae, tribe Lactuceae is capable of synthesizing sesquiterpene lactones of which ixerin F is the main compound. The culture was characterized on growth (by dissimilation rates, on ixerin F accumulation (by RP-HPLC and on some enzyme activities involved in early steps of terpenoid biosynthesis. Acetoacetyl-coenzyme A thiolase (AACT, E.C. 2.3.1.9 and 3S-hydroxy-3-methylglutaryl-coenzyme A synthase (HMGS, E.C. 4.13.5 activities of the cells were assayed spectrophotometrically. HMGS activity increased during the culture period and reached a maximum during the stationary phase (190 pkat/mg protein, while AACT showed relatively high level of activity throughout the growth cycle, with transient decrease at the logarithmic growth phase and the beginning of stationary phase. Ixerin F accumulated inside the cells and the maximum concentration of 0.08% (on dry weight basis was found in the early stationary phase of the growth cycle of the culture.

  12. Changes in auxin level in the course of growth of a sunflower crown-gall suspension culture

    Directory of Open Access Journals (Sweden)

    Zofia Chirek

    2014-01-01

    Full Text Available The auxin level in the cell mass and culture medium was determined by means of the Avena straight caleoptile test in various periods of the suspension culture cycle of the sunflower crown-gall tumour. The investigations were performed in the course of the zero passage (PO and first one (Pl, differing in their time of duration of maximum growth and its intensity. In both passages the intra- and extra-cellular auxin levels reach values of the same order. At the beginning of the maximal growth phase the activity corresponding to IAA in the cells prevails over that of the other auxin-like compounds. This disproportion diminishes with further development of the culture, and with the beginning of the stationary phase the cellular IAA level is lower than that of the remaining auxin-like compounds. The short phase of maximal growth (PO occurs with an auxin level decreasing in the cell mass and increasing in the medium, and towards the end of the cycle these levels become equal. During the long phase of maximal growth (Pl the total amount of auxins in the cells increases and is 2-3 times higher than in the medium, whereas IAA in the cells remains at a constant level. These results suggest that the participation of IAA in the intracellular pool of auxin-like substances is decisive for the mitotic activity of the cells and maintenance of growth in the culture.

  13. The Effect of Plant Growth Regulators and Different Explants on the Response of Tissue Culture and Cell Suspension Cultures of German Chamomile (Matricaria chamomilla L.

    Directory of Open Access Journals (Sweden)

    L. Koohi,

    2014-07-01

    Full Text Available German chamomile (Matricaria chamomilla L. is one of the most important medicinal plants that its essential oils used in different medicinal industries. In this study which was carried out in 2013 growing season at the Faculty of Agricultural Sciences of the University of Mohaghegh Ardabili, the in vitro response of leaf and hypocotyl explants of German Chamomile in B5 medium supplemented with different levels of plant growth regulators including 2,4-D, naphthalene acetic acid (NAA, kinetin and 6-benzylaminopurine (BAP were investigated in a factorial experiment based on completely randomized design (CRD.In addition, cell suspension cultures were established and characterized. Hypocotyl and leaf explants exhibited cell proliferation and produced callus within 1-2 weeks. The highest fresh weight of the callus (264.1 mg was produced by leaf explants in the medium supplemented with 0.5 mg/l 2,4-D and 1 mg/l BAP. However, the leaf explants cultured on medium containing 1.5 mg/l 2,4-D showed the lowest cell proliferation and callus yield (40.42 mg. The highest percentage of root induction from leaf explants (58.73% was observed on the medium containing 4 mg/l 2,4-D and 1 mg/l Kin, and from hypocotyl explants (48.61% was observed on medium supplemented with 1.5 mg/l NAA. The 42.22% of calli derived from hypocotyl explants on B5 medium supplemented with 4 mg/l NAA and 3 mg/l BAP, were friable. Cell suspension cultures of German chamomile were established by transferring of hypocotyl-derived friable calli into the MS medium supplemented with 1.5 mg/l 2,4-D and 1 mg/l kinetin. The growth curve of cell proliferations started 4 days after culture and continued to grow until day 13th, where the cells entered stationary phase.

  14. Biophysical monitoring of cell cultures for quality assessment utilizing digital holographic microscopy

    Science.gov (United States)

    Kastl, Lena; Isbach, Michael; Dirksen, Dieter; Schnekenburger, Jürgen; Kemper, Björn

    2017-06-01

    Quality and reproducibility of cell-based assays strongly depend on the quality of the underlying cell culture which is influenced by various parameters like nutrient and growth factor availability, buffer conditions, subculture routines and optimal cell concentrations. Thus, methods for accurate assessment of objective cell parameters that characterize a specific cell line and detect global changes in cell culture are highly desirable. During the past years, quantitative phase imaging has been recognized as a promising tool for quantitative label-free live cell analysis. We demonstrate the utilization of quantitative phase imaging with digital holographic microscopy (DHM) to quantify the impact of cell culture conditions on single cells using a pancreatic tumor cell model. Label-free quantitative phase imaging of detached cells in suspension is performed by Michelson interferometer-based self-interference DHM. The quantitative phase images of the cells are analyzed for refractive index, volume and dry mass. We show that the evaluation of quantitative DHM phase images allows to extract absolute biophysical cellular parameters that are related to cell layer confluence states. In summary, the results of our study demonstrate that DHM is capable for label-free imaging cytometry with novel biophysical data sets that are acquired with minimum sample preparation for sophisticated monitoring of cell morphology alterations that are related to changes of cell culture conditions.

  15. Violin mode amplitude glitch monitor for the presence of excess noise on the monolithic silica suspensions of GEO 600

    Energy Technology Data Exchange (ETDEWEB)

    Sorazu, B; Strain, K A; Heng, I S; Kumar, R, E-mail: b.sorazu@physics.gla.ac.u [Department of Physics and Astronomy, University of Glasgow, University Avenue, Glasgow G12 8QQ (United Kingdom)

    2010-08-07

    Non-Gaussian features of data from gravitational wave detectors are of interest as unpredictable 'glitches' limit the sensitivity of searches for many kinds of signal. We consider events due to non-random excitations of the test masses and their suspension fibres. These events could, for example, be related to acoustic emissions in the fibres due to the presence and propagation of cracks or another type of structural perturbation, and they would generate excess noise above the Gaussian background, which matches the level expected due to thermal noise. We look for excess noise in the fundamental violin modes of the monolithic silica suspension fibres of GEO 600. We describe the algorithm used to monitor the violin mode amplitude for glitches, present our results and consider how these may be applied to advanced detectors. The conclusion of our analysis is that no excess noise above what was considered to be thermal noise was observed for several days of h(t) data analysed at the frequency of the selected violin modes.

  16. Violin mode amplitude glitch monitor for the presence of excess noise on the monolithic silica suspensions of GEO 600

    International Nuclear Information System (INIS)

    Sorazu, B; Strain, K A; Heng, I S; Kumar, R

    2010-01-01

    Non-Gaussian features of data from gravitational wave detectors are of interest as unpredictable 'glitches' limit the sensitivity of searches for many kinds of signal. We consider events due to non-random excitations of the test masses and their suspension fibres. These events could, for example, be related to acoustic emissions in the fibres due to the presence and propagation of cracks or another type of structural perturbation, and they would generate excess noise above the Gaussian background, which matches the level expected due to thermal noise. We look for excess noise in the fundamental violin modes of the monolithic silica suspension fibres of GEO 600. We describe the algorithm used to monitor the violin mode amplitude for glitches, present our results and consider how these may be applied to advanced detectors. The conclusion of our analysis is that no excess noise above what was considered to be thermal noise was observed for several days of h(t) data analysed at the frequency of the selected violin modes.

  17. Violin mode amplitude glitch monitor for the presence of excess noise on the monolithic silica suspensions of GEO 600

    Science.gov (United States)

    Sorazu, B.; Strain, K. A.; Heng, I. S.; Kumar, R.

    2010-08-01

    Non-Gaussian features of data from gravitational wave detectors are of interest as unpredictable 'glitches' limit the sensitivity of searches for many kinds of signal. We consider events due to non-random excitations of the test masses and their suspension fibres. These events could, for example, be related to acoustic emissions in the fibres due to the presence and propagation of cracks or another type of structural perturbation, and they would generate excess noise above the Gaussian background, which matches the level expected due to thermal noise. We look for excess noise in the fundamental violin modes of the monolithic silica suspension fibres of GEO 600. We describe the algorithm used to monitor the violin mode amplitude for glitches, present our results and consider how these may be applied to advanced detectors. The conclusion of our analysis is that no excess noise above what was considered to be thermal noise was observed for several days of h(t) data analysed at the frequency of the selected violin modes.

  18. Chemical Elicitor-Induced Modulation of Antioxidant Metabolism and Enhancement of Secondary Metabolite Accumulation in Cell Suspension Cultures of Scrophularia kakudensis Franch

    Directory of Open Access Journals (Sweden)

    Abinaya Manivannan

    2016-03-01

    Full Text Available Scrophularia kakudensis is an important medicinal plant with pharmaceutically valuable secondary metabolites. To develop a sustainable source of naturaceuticals with vital therapeutic importance, a cell suspension culture was established in S. kakudensis for the first time. Friable calli were induced from the leaf explants cultured on a Murashige and Skoog (MS medium containing 3.0 mg·L−1 6-benzyladenine (BA in a combination with 2 mg·L−1 2,4-dichlorophenoxy acetic acid (2,4-D. From the callus cultures, a cell suspension culture was initiated and the cellular differentiation was investigated. In addition, the effect of biotic elicitors such as methyl jasmonate (MeJa, salicylic acid (SA, and sodium nitroprusside (SNP on the accumulation of secondary metabolites and antioxidant properties was demonstrated. Among the elicitors, the MeJa elicited the accumulation of total phenols, flavonoids, and acacetin, a flavonoid compound with multiple pharmaceutical values. Similarly, the higher concentrations of the MeJa significantly modulated the activities of antioxidant enzymes and enhanced the scavenging potentials of free radicals of cell suspension extracts. Overall, the outcomes of this study can be utilized for the large scale production of pharmaceutically important secondary metabolites from S. kakudensis through cell suspension cultures.

  19. Chemical Elicitor-Induced Modulation of Antioxidant Metabolism and Enhancement of Secondary Metabolite Accumulation in Cell Suspension Cultures of Scrophularia kakudensis Franch.

    Science.gov (United States)

    Manivannan, Abinaya; Soundararajan, Prabhakaran; Park, Yoo Gyeong; Jeong, Byoung Ryong

    2016-03-18

    Scrophularia kakudensis is an important medicinal plant with pharmaceutically valuable secondary metabolites. To develop a sustainable source of naturaceuticals with vital therapeutic importance, a cell suspension culture was established in S. kakudensis for the first time. Friable calli were induced from the leaf explants cultured on a Murashige and Skoog (MS) medium containing 3.0 mg·L(-1) 6-benzyladenine (BA) in a combination with 2 mg·L(-1) 2,4-dichlorophenoxy acetic acid (2,4-D). From the callus cultures, a cell suspension culture was initiated and the cellular differentiation was investigated. In addition, the effect of biotic elicitors such as methyl jasmonate (MeJa), salicylic acid (SA), and sodium nitroprusside (SNP) on the accumulation of secondary metabolites and antioxidant properties was demonstrated. Among the elicitors, the MeJa elicited the accumulation of total phenols, flavonoids, and acacetin, a flavonoid compound with multiple pharmaceutical values. Similarly, the higher concentrations of the MeJa significantly modulated the activities of antioxidant enzymes and enhanced the scavenging potentials of free radicals of cell suspension extracts. Overall, the outcomes of this study can be utilized for the large scale production of pharmaceutically important secondary metabolites from S. kakudensis through cell suspension cultures.

  20. Safety Culture Monitoring: How to Assess Safety Culture in Real Time?

    International Nuclear Information System (INIS)

    Zronek, B.; Maryska, J.; Treslova, L.

    2016-01-01

    Do you know what is current level of safety culture in your company? Are you able to follow trend changes? Do you know what your recent issues are? Since safety culture is understood as vital part of nuclear industry daily life, it is crucial to know what the current level is. It is common to perform safety culture survey or ad hoc assessment. This contribution shares Temelin NPP, CEZ approach how to assess safety culture level permanently. Using behavioral related outputs of gap solving system, observation program, dedicated surveys, regulatory assessment, etc., allows creating real time safety culture monitoring without the need to perform any other activities. (author)

  1. Regeneration of transgenic cassava plants (Manihot esculenta Crantz) from microbombarded embryogenic suspension cultures.

    Science.gov (United States)

    Schöpke, C; Taylor, N; Cárcamo, R; Konan, N K; Marmey, P; Henshaw, G G; Beachy, R N; Fauquet, C

    1996-06-01

    A protocol was established for the introduction of DNA into embryogenic suspension-derived tissues of cassava via microparticle bombardment, for the selection of genetically transformed cells, and for the regeneration of fully transgenic plants from these cells. The plasmid DNA used for bombardment contained a gene encoding neomycin phosphotransferase (nptII) and a gene encoding beta-glucuronidase (uidA). Selection of bombarded tissue with paromomycin resulted in the establishment of putative transgenic embryogenic calli. In most of these calli, beta-glucuronidase was detected histochemically. Molecular analysis of paromomycin-resistant embryogenic calli and of plants regenerated from these calli, confirmed the stable integration of bombarded DNA into the cassava genome.

  2. Regulation of Cytoplasmic and Vacuolar Volumes by Plant Cells in Suspension Culture

    DEFF Research Database (Denmark)

    Owens, Trevor; Poole, Ronald J

    1979-01-01

    by cytoplasm is roughly correlated with protein content, but shows no correlation with cell size or with intracellular concentrations of K or Na. The most striking observation is that the growth of cytoplasmic volume for the culture as a whole appears to be constant throughout the culture cycle, despite...

  3. Isoferritins in rat Kupffer cells, hepatocytes, and extrahepatic macrophages. Biosynthesis in cell suspensions and cultures in response to iron

    International Nuclear Information System (INIS)

    Doolittle, R.L.; Richter, G.W.

    1981-01-01

    Cultures of Kupffer cells and of hepatocytes, prepared from single rat livers, synthesized ferritin protein equally efficiently. In culture but not in suspension, both sorts of cells responded significantly to stimulation with iron by increased ferritin synthesis. As determined by isoelectric focusing, the isoferritin profiles of newly synthesized 14 -labeled Kupffer cell and hepatocyte ferritin were identical, each having three bands. However, unlabeled ferritin, extracted from nonparenchymal liver cells (mainly Kupffer and endothelial cells) of iron-loaded rats, contained an acidic isoferritin that was not present in hepatocyte ferritin. Investigation of ferritin synthesis in cultured peritoneal and alveolar macrophages yielded similar results. The isofocusing profile of newly synthesized peritoneal macrophage ferritin was indistinguishable from the profile of fresh Kupffer cell or hepatocyte ferritin. Thus, the three isoferritins common to Kupffer cells, hepatocytes, and extrahepatic macrophages are neither cell- nor tissue-specific. However, modifications on intracellular storage may affect the isofocusing properties. The findings, although consistent with the LnH24-n subunit model of ferritin protein, indicate identical restrictive genomic control of the H:L ratios in these sorts of cells. Further, they make it probable that Kupffer cell ferritin iron, originating by endogenous synthesis, is the principal source of Kupffer cell hemosiderin iron

  4. Hydrodynamic stress induces monoterpenoid oxindole alkaloid accumulation by Uncaria tomentosa (Willd) D. C. cell suspension cultures via oxidative burst.

    Science.gov (United States)

    Trejo-Tapia, Gabriela; Sepúlveda-Jiménez, Gabriela; Trejo-Espino, José Luis; Cerda-García-Rojas, Carlos M; de la Torre, Mayra; Rodríguez-Monroy, Mario; Ramos-Valdivia, Ana C

    2007-09-01

    Uncaria tomentosa cell suspension cultures were grown in a 2-L stirred tank bioreactor operating at a shear rate gamma(.)(avg)=86 s(-1). The cultures showed an early monophasic oxidative burst measured as H2O2 production (2.15 micromol H2O2 g(-1) dw). This response was followed by a transient production of monoterpenoid oxindole alkaloids (178 +/- 40 microg L(-1) at 24 h). At the stationary phase (144 h), the increase of the shear rate gamma(.)(avg) up to 150 s(-1) and/or oxygen tension up to 85% generated H2O2, restoring oxindole alkaloid production. U. tomentosa cells cultured in Erlenmeyer flasks also exhibited the monophasic oxidative burst but the H2O2 production was 16-fold lower and the alkaloids were not detected. These cells exposed to H2O2 generated in situ produced oxindole alkaloids reaching a maximum of 234 +/- 40 microg L(-1). A positive correlation was observed between the oxindole alkaloid production and the endogenous H2O2 level. On the other hand, addition of 1 microM diphenyleneiodonium (NAD(P)H oxidase inhibitor) or 10 microM sodium azide (peroxidases inhibitor) reduced both H2O2 production and oxindole alkaloids build up, suggesting that these enzymes might play a role in the oxidative burst induced by the hydrodynamic stress.

  5. Role of Changes in Cell Fatty Acids Composition in the Increasing of Frost Resistance of Winter Wheat Suspension Culture

    Directory of Open Access Journals (Sweden)

    I.V. Lyubushkina

    2013-11-01

    Full Text Available Influences of low temperatures (4 and 8 ° С on the frost tolerance and fatty acid compositions of cells in a winter wheat suspension culture have been studied. It has been found that treatment of the culture with 4 °C (7 days did not protect cells from subsequent freezing temperature action (-8 °С, 6 h and was not accompanied significant changes in the fatty acid composition. On the contrary, the treatment of the culture with the temperature 8 °C (7 days prevented the death caused by freezing temperature and the content of saturated fatty acids decreased: pentadecanoic acid (by 35,0%, palmitic acid (by 19,9% and stearic acid (by 65,4%, and the content of α-linolenic acid increased by 94%. That was the cause of the double bond index (DBI increase by 16%. The role of fatty acids composition changes in the process of increasing frost tolerance in plants are discussed.

  6. Effects of aluminum on DNA synthesis, cellular polyamines, polyamine biosynthetic enzymes and inorganic ions in cell suspension cultures of a woody plant, Catharanthus roseus

    Science.gov (United States)

    Rakesh Minocha; Subhash C. Minocha; Stephanie L. Long; Walter C. Shortle

    1992-01-01

    Increased aluminum (Al) solubility in soil waters due to acid precipitation has aroused considerable interest in the problem of Al toxicity in plants. In the present study, an in vitro suspension culture system of Catharanthus roseus (L.) G. Don was used to analyze the effects of aluminum on several biochemical processes in these cells. The aliphatic...

  7. Changes in phytochelatins and their biosynthetic intermediates in red spruce (Picea rubens Sarg.) cell suspension cultures under cadmium and zinc stress

    Science.gov (United States)

    P. Thangavel; Stephanie Long; Rakesh Minocha

    2007-01-01

    Cell suspension cultures of red spruce (Picea rubens Sarg.) were selected to study the effects of cadmium (Cd) and zinc (Zn) on phytochelatins (PCs) and related metabolites after 24 h exposure. The PC2 and its precursor, γ-glutamylcysteine (γ-EC) increased two to fourfold with Cd concentrations ranging from 12...

  8. Quinoline Alkaloids in Suspension Cultures of Cinchona ledgeriana Treated with Various Substances

    Directory of Open Access Journals (Sweden)

    DIAH RATNADEWI

    2010-12-01

    Full Text Available Cinchona alkaloids are in extensive uses, not only for drugs but also for soft drink industries. They are harvested from the bark of trees Cinchona spp. after certain ages and therefore are available over a limited time. Cell culture is an alternative way to continuously produce such secondary metabolites in a much shorter time. Various substances were added in the normal growth media to promote quinoline alkaloids production by cell cultures of Cinchona ledgeriana. At the sixth week of culture, quinine and cinchonine contents were suppressed by paclobutrazol (PBZ, abscisic acid (ABA, or even by precursor tryptophan, while cinchonidine content was enhanced by 0.2 mg/l tryptophan to 43 fold of that produced by untreated cells (2.8% dry weight. At the seventh week of culture, the production of quinine and quinidine started to grow whereas the production of cinchonine and cinchonidine tended to decrease. An addition of 5 mg/l PBZ to culture media yielded the highest level of total quinine/quinidine after seven weeks, e.g. quinine 11 times more abundant and quinidine 23 fold higher compared to the untreated cells. Particularly the level of quinine which is the most demanded for medical and industrial purposes still need to be improved to approach to or even higher than that of extracted from the conventional source.

  9. Selection and optimization of transfection enhancer additives for increased virus-like particle production in HEK293 suspension cell cultures.

    Science.gov (United States)

    Cervera, Laura; Fuenmayor, Javier; González-Domínguez, Irene; Gutiérrez-Granados, Sonia; Segura, Maria Mercedes; Gòdia, Francesc

    2015-12-01

    The manufacturing of biopharmaceuticals in mammalian cells typically relies on the use of stable producer cell lines. However, in recent years, transient gene expression has emerged as a suitable technology for rapid production of biopharmaceuticals. Transient gene expression is particularly well suited for early developmental phases, where several potential therapeutic targets need to be produced and tested in vivo. As a relatively new bioprocessing modality, a number of opportunities exist for improving cell culture productivity upon transient transfection. For instance, several compounds have shown positive effects on transient gene expression. These transfection enhancers either facilitate entry of PEI/DNA transfection complexes into the cell or nucleus or increase levels of gene expression. In this work, the potential of combining transfection enhancers to increase Gag-based virus-like particle production levels upon transfection of suspension-growing HEK 293 cells is evaluated. Using Plackett-Burman design of experiments, it is first tested the effect of eight transfection enhancers: trichostatin A, valproic acid, sodium butyrate, dimethyl sulfoxide (DMSO), lithium acetate, caffeine, hydroxyurea, and nocodazole. An optimal combination of compounds exhibiting the highest effect on gene expression levels was subsequently identified using a surface response experimental design. The optimal consisted on the addition of 20 mM lithium acetate, 3.36 mM valproic acid, and 5.04 mM caffeine which increased VLP production levels 3.8-fold, while maintaining cell culture viability at 94%.

  10. Effects of exogenous growth regulators on cell suspension culture of yin-hong grape (vitis vinifera l.) and establishment of the optimum medium

    International Nuclear Information System (INIS)

    Chao, Y.; Feng, J.C.; Yan, W.Y.; Xiao, Y.; Jun, Y.Y

    2015-01-01

    Callus induced by stem of Yin-hong grape (Vitis vinifera L.) was used as materials and B5 medium as basic medium. The major growth parameters of cell suspension cultures with various levels of 1-Naphthaleneacetic acid (NAA) and 6-Benzyl aminopurine (6-BA) were investigated to provide a basis for the optimum medium of suspension cell cultures of Yin-hong grape regarding cell number, packed cell volume (PCV), dry cell weight (DCW), cell viability, and morphology. All data were analysed by of two-way analysis of variance (ANOVA). Results showed that the treatment of 6-BA and NAA would effect the cell growth dynamics, probably causing logarithmic phase in advance at higher levels of 6-BA. Different concentration of 6-BA and NAA had significant effects on cells number, PCV, DCW and viability (p<0.05), while no-significant effect was observed on the cells morphology. The optimum medium for suspension cell cultures of Yin-hong grape was identified as B5+1.5 mg/L6-BA+1.5 mg/LNAA+ 250 mg/L casein hydrolysate + 30 g/L sucrose. With the optimum medium, the maximum number of suspension cells after the logarithmic growth phase was 34.78 * 108 / mL, the highest cell viability reached 86.45%.; DCW reached 3.84 g/L and PCV reached 0.092 mL/mL after eight days cultivating. (author)

  11. Growth and production optimization of tropane alkaloids in Datura stramonium cell suspension culture.

    Science.gov (United States)

    Iranbakhsh, A R; Oshagi, M A; Ebadi, M

    2007-04-15

    Abstract: A number of physicochemical conditions such different concentration of glucose, sucrose, potassium nitrate, ammonium nitrate, calcium chloride and temperatures were tested to optimize growth and production of tropane alkaloids from Datura stramonium (Solanaceae) plants. Cell suspension from semi-clear calli of leave explants developed in MS medium containing kinetin (0.5 mg L(-1)) and NAA (2 mg L(-1)) hormones was used to measure biomass and total alkaloids and comparison of treatments. The results showed that 30 and 40 g L(-1) glucose led to the highest level of alkaloids and biomass productions, respectively. 20 and 40 g L(-1) sucrose concentrations resulted in order the most rates of alkaloids and biomass productions. The results showed that increasing of nitrate concentration led to the reduction of the alkaloids. The best concentration of potassium nitrate for the production of tropane alkaloids and biomass were in order 9.4 and 3.76 mM. Also it was evinced that the optimized concentration of ammonium nitrate for alkaloids production was 10.3 mM and for the biomass was 41.22 mM. The best concentration of calcium chloride for growth and production of the alkaloids was 7.92 mM. Testing different temperature specified that the best condition for production of the alkaloids was 20 degrees C whereas it was 25 degrees C for biomass production. The results of this study could be recommended to farmers involved in production of D. stramonium for tropain alkaloids at industrial and semi-industrial scales.

  12. Role of polyamines in DNA synthesis of Catharanthus roseus cells grown in suspension culture

    Science.gov (United States)

    Rakesh Minocha; Subhash C. Minocha; Atsushi Komamine; Walter C. Shortle

    1990-01-01

    The requirement for polyamines in the proliferation of cells was first demonstrated in bacteria (3). While significant progress has been made in this field using animal cell cultures, only preliminary studies have been reported with plant tissues. Serafini-Fracassini et al. (9) showed a marked increase in polyamine synthesis early during the G 1 phase, concomitant with...

  13. Establishment of Cell Suspension Culture and Plant Regeneration in Abrus precatorius L., a Rare Medicinal Plant

    Directory of Open Access Journals (Sweden)

    Mohammad Serajur RAHMAN

    2012-02-01

    Full Text Available A new protocol has been developed for cell culture and in vitro regeneration of Abrus precatorius that holds enormous potentiality for preparation of medicines. In vitro grown calli were cultured in Murashige and Skoog (MS liquid media in agitated condition fortified with 0.5 mg/l 6-Benzylaminopurine. Growth curve of cells revealed that the cells continued to grow until 12 days of culture and got the highest peak from day 6-8. Isolated cell was found to produce highest 8.2% calli when suspended on MS medium supplemented with 0.5 mg/l 6-Benzylaminopurine and 0.1 mg/l 1-Naphthaleneacetic acid. Callus derived from single cell produced highest number of embryo (25-28% cultured on MS medium fortified with 2.0 mg/l 6-Benzylaminopurine and 0.2 mg/l 1-Naphthaleneacetic acid. The bipolar embryos were selected and optimum shoot formation was recorded on MS medium supplemented with 2.0 mg/l 6-Benzylaminopurine and 0.1 mg/l 1-Naphthaleneacetic acid. The optimum root induction was noticed in MS medium supplemented with 1.0 mg/l 3-Indolebutyric acid. Rooted plantlets were successfully transferred to potting soil and acclimatized to outdoor conditions.

  14. Effects of 60Co γ-rays irradiation on cell growth and alkaloid accumulation of protocorm-like bodies in suspension cultures from Dendrobium huoshanense

    International Nuclear Information System (INIS)

    Hong Sali; Jin Qing; Huang Bei; Cai Yongping; Lin Yi

    2009-01-01

    Protocorm-like bodies (PLBs) in suspension cultures from Dendrobium huoshanense were irradiated by 60 Co γ-rays at doses of 5, 10, 20 and 30Gy, and alkaloid accumulation of PLBs was studied. The results showed that 60 Co γ-rays irradiation could improve the alkaloid content of PLBs, and the suitable dose was 10Gy. The fresh weight of 10Gy irradiated PLBs was 26.54g/flask, and the alkaloid content was 0.035% on the 36th day. The medium pH and electric conductivity of 10Gy irradiated PLBs changed slightly during the suspension culture period. The results suggested such cultural environment was suitable for PLBs growth continuely. Results also showed that 60 Co γ-rays irradiation could increase the activities of POD, SOD, CAT, PAL and decrease the activity of PPO, these were responsible for the improvement of cell growth and alkaloid accumulation in PLBs. (authors)

  15. Effect of salts (NaCl and Na2CO3) on callus and suspension culture of Stevia rebaudiana for Steviol glycoside production.

    Science.gov (United States)

    Gupta, Pratibha; Sharma, Satyawati; Saxena, Sanjay

    2014-03-01

    Steviol glycosides are natural non-caloric sweeteners which are extracted from the leaves of Stevia rebaudiana plant. Present study deals the effect of salts (NaCl and Na2CO3) on callus and suspension culture of Stevia plant for steviol glycoside (SGs) production. Yellow-green and compact calli obtained from in vitro raised Stevia leaves sub-cultured on MS medium supplemented with 2.0 mg l(-1) NAA and different concentrations of NaCl (0.05-0.20%) and Na2CO3 (0.0125-0.10%) for 2 weeks, and incubated at 24 ± 1 °C and 22.4 μmol m(-2) s(-1) light intensity provided by white fluorescent tubes for 16 h. Callus and suspension biomass cultured on salts showed less growth as well as browning of medium when compared with control. Quantification of SGs content in callus culture (collected on 15th day) and suspension cultures (collected at 10th and 15th days) treated with and without salts were analyzed by HPLC. It was found that abiotic stress induced by the salts increased the concentration of SGs significantly. In callus, the quantity of SGs got increased from 0.27 (control) to 1.43 and 1.57% with 0.10% NaCl, and 0.025% Na2CO3, respectively. However, in case of suspension culture, the same concentrations of NaCl and Na2CO3 enhanced the SGs content from 1.36 (control) to 2.61 and 5.14%, respectively, on the 10th day.

  16. Reactive oxygen and nitrogen (ROS and RNS) species generation and cell death in tomato suspension cultures--Botrytis cinerea interaction.

    Science.gov (United States)

    Pietrowska, E; Różalska, S; Kaźmierczak, A; Nawrocka, J; Małolepsza, U

    2015-01-01

    This article reports events connected to cell survival and Botrytis cinerea infection development in cell suspension cultures of two tomato cultivars which show different levels of susceptibility to the pathogen: cv. Corindo (more susceptible) and cv. Perkoz (less susceptible). In parallel changes in reactive oxygen (ROS) and nitrogen (RNS) species generation and in S-nitrosoglutathione reductase (GSNOR) activity were studied. In vivo staining methods with acridine orange (AO) and ethidium bromide (EB) as well as fluorescent microscopy were used to assess tomato and B. cinerea cells death. The biochemical studies of ROS and RNS concentrations in plant cell extract were complemented by in vivo ROS and nitric oxide (NO) imaging using nitro blue tetrazolium (NBT), diaminobenzidine (DAB) and diaminofluorescein diacetate (DAF-DA) staining methods, and confocal microscope technique. B. cinerea infection proceeded slower in Perkoz cell cultures. It was evidenced by measuring the pathogen conidia germination and germination tube development in which nuclei revealing cell death dominated. Two different types of tomato cell death were observed: cells with necrotic nuclei dominated in Corindo whereas in Perkoz cells with characteristic of vacuolar death type prevailed. In Perkoz cells, constitutive levels of NO and S-nitrosothiols (SNO) were significantly higher and hydrogen peroxide (H₂O₂) and superoxide anion (O₂(-)) concentrations were slightly higher as compared with Corindo cells. Moreover, increases in these molecule concentrations as a result of B. cinerea inoculation were observed in both, Perkoz and Corindo cell cultures. The enzymatic GSNOR activity seems to be an important player in controlling the SNO level in tomato cells. Involvements of the studied compounds in molecular mechanisms of tomato resistance to B. cinerea are discussed in the paper.

  17. Identification and expression analysis of methyl jasmonate responsive ESTs in paclitaxel producing Taxus cuspidata suspension culture cells

    Directory of Open Access Journals (Sweden)

    Lenka Sangram K

    2012-04-01

    Full Text Available Abstract Background Taxol® (paclitaxel promotes microtubule assembly and stabilization and therefore is a potent chemotherapeutic agent against wide range of cancers. Methyl jasmonate (MJ elicited Taxus cell cultures provide a sustainable option to meet the growing market demand for paclitaxel. Despite its increasing pharmaceutical importance, the molecular genetics of paclitaxel biosynthesis is not fully elucidated. This study focuses on identification of MJ responsive transcripts in cultured Taxus cells using PCR-based suppression subtractive hybridization (SSH to identify genes involved in global pathway control. Results Six separate SSH cDNA libraries of paclitaxel-accumulating Taxus cuspidata P991 cell lines were constructed at three different post-elicitation time points (6h, 18h and 5 day to identify genes that are either induced or suppressed in response to MJ. Sequencing of 576 differentially screened clones from the SSH libraries resulted in 331 unigenes. Functional annotation and Gene Ontology (GO analysis of up-regulated EST libraries showed enrichment of several known paclitaxel biosynthetic genes and novel transcripts that may be involved in MJ-signaling, taxane transport, or taxane degradation. Macroarray analysis of these identified genes unravelled global regulatory expression of these transcripts. Semi-quantitative RT-PCR analysis of a set of 12 candidate genes further confirmed the MJ-induced gene expression in a high paclitaxel accumulating Taxus cuspidata P93AF cell line. Conclusions This study elucidates the global temporal expression kinetics of MJ responsive genes in Taxus suspension cell culture. Functional characterization of the novel genes identified in this study will further enhance the understanding of paclitaxel biosynthesis, taxane transport and degradation.

  18. Identification and expression analysis of methyl jasmonate responsive ESTs in paclitaxel producing Taxus cuspidata suspension culture cells

    Science.gov (United States)

    2012-01-01

    Background Taxol® (paclitaxel) promotes microtubule assembly and stabilization and therefore is a potent chemotherapeutic agent against wide range of cancers. Methyl jasmonate (MJ) elicited Taxus cell cultures provide a sustainable option to meet the growing market demand for paclitaxel. Despite its increasing pharmaceutical importance, the molecular genetics of paclitaxel biosynthesis is not fully elucidated. This study focuses on identification of MJ responsive transcripts in cultured Taxus cells using PCR-based suppression subtractive hybridization (SSH) to identify genes involved in global pathway control. Results Six separate SSH cDNA libraries of paclitaxel-accumulating Taxus cuspidata P991 cell lines were constructed at three different post-elicitation time points (6h, 18h and 5 day) to identify genes that are either induced or suppressed in response to MJ. Sequencing of 576 differentially screened clones from the SSH libraries resulted in 331 unigenes. Functional annotation and Gene Ontology (GO) analysis of up-regulated EST libraries showed enrichment of several known paclitaxel biosynthetic genes and novel transcripts that may be involved in MJ-signaling, taxane transport, or taxane degradation. Macroarray analysis of these identified genes unravelled global regulatory expression of these transcripts. Semi-quantitative RT-PCR analysis of a set of 12 candidate genes further confirmed the MJ-induced gene expression in a high paclitaxel accumulating Taxus cuspidata P93AF cell line. Conclusions This study elucidates the global temporal expression kinetics of MJ responsive genes in Taxus suspension cell culture. Functional characterization of the novel genes identified in this study will further enhance the understanding of paclitaxel biosynthesis, taxane transport and degradation. PMID:22530557

  19. Comparison of the Production of Recombinant Protein in Suspension Culture of CHO Cells in Spinner Flask and Shake Flask System

    Directory of Open Access Journals (Sweden)

    S.N.Z Zainul Abidin

    2011-12-01

    Full Text Available Chinese hamster ovary (CHO cells have been most widely used as the production host for the commercial production of biopharmaceuticals product. They have been extensively studied and developed, and today provide a stable platform for producing monoclonal antibodies and recombinant proteins. This study was focusing on comparison of suspension culture system by using spinner flask and shake flask for the growth and production of recombinant protein in CHO cell line. The CHO cells were transfected with an expression of DNA plasmid containing lac Z gene which codes for β-galactosidase. The recombinant genes in these CHO cells and the β-galactosidase expressing cells were adapted to suspension culture. The agitation speed for both spinner and shake flask were adjusted accordingly. The experiments were carried out in duplicate and samples were taken for cell count, determination of glucose consumption, lactate production and protein level by using biochemical assay. The result showed that, the cell growth in spinner flask is more favorable then in shake flask. The cell concentration in spinner flask is 58% higher than in shake flask. On the other hand, specific activity of β-galactosidase is 25% higher in spinner flask compared to shake flask, at the same agitation speed.ABSTRAK: Sel ovari hamster China (Chinese hamster ovary (CHO digunakan secara meluas dalam hos pembiakan untuk tujuan komersil produk biofarmaseutikal. Ia telah dikaji dan dibangunkan secara ekstensif, dan kini ia menyediakan landasan yang stabil untuk penghasilan antibodi monoklon dan protein rekombinan. Kajian ini memfokuskan tentang penghasilan protein rekombinan menggunakan kultur ampaian sel CHO di dalam kelalang putar dan kelalang goncang. Sel CHO dimasukkan dengan plasmid DNA yang mengandungi gen lac Z yang juga memberikan kod untuk β-galaktosidase. Sel CHO β-galaktosidase-terungkap dimasukkan ke dalam kultur ampaian. Kelajuan agitasi untuk kedua-dua kelalang putar

  20. Chloride secretagogues stimulate inositol phosphate formation in shark rectal gland tubules cultured in suspension

    International Nuclear Information System (INIS)

    Ecay, T.W.; Valentich, J.D.

    1991-01-01

    Neuroendocrine activation of transepithelial chloride secretion by shark rectal gland cells is associated with increases in cellular cAMP, cGMP, and free calcium concentrations. We report here on the effects of several chloride secretagogues on inositol phosphate formation in cultured rectal gland tubules. Vasoactive intestinal peptide (VIP), atriopeptin (AP), and ionomycin increase the total inositol phosphate levels of cultured tubules, as measured by ion exchange chromatography. Forskolin, a potent chloride secretagogue, has no effect on inositol phosphate formation. The uptake of 3 H-myo-inositol into phospholipids is very slow, preventing the detection of increased levels of inositol trisphosphate. However, significant increases in inositol monophosphate (IP1) and inositol biphosphate (IP2) were measured. The time course of VIP- and AP-stimulated IP1 and IP2 formation is similar to the effects of these agents on the short-circuit current responses of rectal gland monolayer cultures. In addition, aluminum fluoride, an artificial activator of guanine nucleotide-binding proteins, stimulates IP1 and IP2 formation. We conclude that rectal gland cells contain VIP and AP receptors coupled to the activation of phospholipase C. Coupling may be mediated by G-proteins. Receptor-stimulated increases in inositol phospholipid metabolism is one mechanism leading to increased intracellular free calcium concentrations, an important regulatory event in the activation of transepithelial chloride secretion by shark rectal gland epithelial cells

  1. INL Cultural Resource Monitoring Report for FY 2015

    Energy Technology Data Exchange (ETDEWEB)

    Pace, Brenda Ringe [Idaho National Lab. (INL), Idaho Falls, ID (United States); Olson, Christina Liegh [Idaho National Lab. (INL), Idaho Falls, ID (United States); Gilbert, Hollie Kae [Idaho National Lab. (INL), Idaho Falls, ID (United States); Holmer, Marie Pilkington [Idaho National Lab. (INL), Idaho Falls, ID (United States)

    2015-10-01

    This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during fiscal year (FY) 2015. Throughout the year, 67 total monitoring visits were completed, with several especially sensitive resources visited on more than one occasion. Overall, FY 2015 monitoring included surveillance of the following 49 individual cultural resource localities: three locations with human remains, one of which is also a cave; nine additional caves; twenty prehistoric archaeological sites; five historic archaeological sites; two historic trails; Experimental Breeder Reactor I (EBR-I), a National Historic Landmark; Aircraft Nuclear Propulsion (ANP) objects located at EBR-I; and eight Arco Naval Proving Ground (NPG) property types. Several INL work processes and projects were also monitored to confirm compliance with original INL CRM recommendations and assess the effects of ongoing work. On two occasions, ground disturbing activities within the boundaries of the Power Burst Facility/Critical Infrastructure Test Range Complex (PBF/CITRC) were observed by INL CRM staff prepared to respond to any additional finds of Native American human remains. Finally, the current location housing INL Archives and Special Collections was evaluated once. Most of the cultural resources monitored in FY 2015 exhibited no adverse impacts, resulting in Type 1 impact assessments. However, Type 2 impacts were noted 13 times. In one case, a portion of a historic trail was graded without prior review or coordination with the INL CRM Office, resulting in impacts to the surface of the trail and one archaeological site. Evidence of unauthorized artifact collection/ looting was also documented at three archaeological sites located along INL powerlines. Federal agents concluded a FY 2012 investigation by filing civil charges and levying fine under the Archaeological Resource Protection Act against one INL employee for this kind

  2. Factorial experimental design for the culture of human embryonic stem cells as aggregates in stirred suspension bioreactors reveals the potential for interaction effects between bioprocess parameters.

    Science.gov (United States)

    Hunt, Megan M; Meng, Guoliang; Rancourt, Derrick E; Gates, Ian D; Kallos, Michael S

    2014-01-01

    Traditional optimization of culture parameters for the large-scale culture of human embryonic stem cells (ESCs) as aggregates is carried out in a stepwise manner whereby the effect of varying each culture parameter is investigated individually. However, as evidenced by the wide range of published protocols and culture performance indicators (growth rates, pluripotency marker expression, etc.), there is a lack of systematic investigation into the true effect of varying culture parameters especially with respect to potential interactions between culture variables. Here we describe the design and execution of a two-parameter, three-level (3(2)) factorial experiment resulting in nine conditions that were run in duplicate 125-mL stirred suspension bioreactors. The two parameters investigated here were inoculation density and agitation rate, which are easily controlled, but currently, poorly characterized. Cell readouts analyzed included fold expansion, maximum density, and exponential growth rate. Our results reveal that the choice of best case culture parameters was dependent on which cell property was chosen as the primary output variable. Subsequent statistical analyses via two-way analysis of variance indicated significant interaction effects between inoculation density and agitation rate specifically in the case of exponential growth rates. Results indicate that stepwise optimization has the potential to miss out on the true optimal case. In addition, choosing an optimum condition for a culture output of interest from the factorial design yielded similar results when repeated with the same cell line indicating reproducibility. We finally validated that human ESCs remain pluripotent in suspension culture as aggregates under our optimal conditions and maintain their differentiation capabilities as well as a stable karyotype and strong expression levels of specific human ESC markers over several passages in suspension bioreactors.

  3. Application of AN Automated Wireless Structural Monitoring System for Long-Span Suspension Bridges

    Science.gov (United States)

    Kurata, M.; Lynch, J. P.; van der Linden, G. W.; Hipley, P.; Sheng, L.-H.

    2011-06-01

    This paper describes an automated wireless structural monitoring system installed at the New Carquinez Bridge (NCB). The designed system utilizes a dense network of wireless sensors installed in the bridge but remotely controlled by a hierarchically designed cyber-environment. The early efforts have included performance verification of a dense network of wireless sensors installed on the bridge and the establishment of a cellular gateway to the system for remote access from the internet. Acceleration of the main bridge span was the primary focus of the initial field deployment of the wireless monitoring system. An additional focus of the study is on ensuring wireless sensors can survive for long periods without human intervention. Toward this end, the life-expectancy of the wireless sensors has been enhanced by embedding efficient power management schemes in the sensors while integrating solar panels for power harvesting. The dynamic characteristics of the NCB under daily traffic and wind loads were extracted from the vibration response of the bridge deck and towers. These results have been compared to a high-fidelity finite element model of the bridge.

  4. Enhanced accumulation of phytosterols and phenolic compounds in cyclodextrin-elicited cell suspension culture of Daucus carota.

    Science.gov (United States)

    Miras-Moreno, Begoña; Almagro, Lorena; Pedreño, M A; Sabater-Jara, Ana Belén

    2016-09-01

    In this work, suspension-cultured cells of Daucus carota were used to evaluate the effect of β-cyclodextrins on the production of isoprenoid and phenolic compounds. The results showed that the phytosterols and phenolic compounds were accumulated in the extracellular medium (15100μgL(-1) and 477.46μgL(-1), respectively) in the presence of cyclodextrins. Unlike the phytosterol and phenolic compound content, β-carotene (1138.03μgL(-1)), lutein (25949.54μgL(-1)) and α-tocopherol (8063.82μgL(-1)) chlorophyll a (1625.13μgL(-1)) and b (9.958 (9958.33μgL(-1)) were mainly accumulated inside the cells. Therefore, cyclodextrins were able to induce the cytosolic mevalonate pathway, increasing the biosynthesis of phytosterols and phenolic compounds, and accumulate them outside the cells. However, in the absence of these cyclic oligosaccharidic elicitors, carrot cells mainly accumulated carotenoids through the methylerythritol 4-phosphate pathway. Therefore, the use of cyclodextrins would allow the extracellular accumulation of both phytosterols and phenolic compounds by diverting the carbon flux towards the cytosolic mevalonate/phenylpropanoid pathway. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  5. Phenylpropanoid Metabolism in Suspension Cultures of Vanilla planifolia Andr. : II. Effects of Precursor Feeding and Metabolic Inhibitors.

    Science.gov (United States)

    Funk, C; Brodelius, P E

    1990-09-01

    Feeding of cinnamic acid and ferulic acid to non-treated and chitosan-treated cell suspension cultures of Vanilla planifolia resulted in the formation of trace amounts of p-hydroxy benzoic acid (5.2 micrograms per gram fresh weight of cells) and vanillic acid (6.4 micrograms per gram fresh weight of cells), respectively. Addition of a 4-hydroxycinnamate: CoA-ligase inhibitor, 3,4-(methylenedioxy)-cinnamic acid (MDCA), resulted in a reduced biosynthesis of ligneous material with a simultaneous significant increased vanillic acid formation (around 75 micrograms per gram fresh weight of cells). A K(1) of 100 micromolar for 4-hydroxycinnamate: CoA-ligase in a crude preparation was estimated for this inhibitor. It is suggested that the conversion of cinnamic acids into benzoic acids does not involve cinnamoyl CoA esters as intermediates. Feeding of (14)C-cinnamic acid and (14)C-ferulic acid to cells treated with MDCA indicate that cinnamic acid, but not ferulic acid, is a precursor of vanillic acid in these cultivated cells of V. planifolia.

  6. Phenylpropanoid Metabolism in Suspension Cultures of Vanilla planifolia Andr. : III. Conversion of 4-Methoxycinnamic Acids into 4-Hydroxybenzoic Acids.

    Science.gov (United States)

    Funk, C; Brodelius, P E

    1990-09-01

    Feeding of 4-methoxycinnamic acid, 3,4-dimethoxycinnamic acid and 3,4,5-trimethoxycinnamic acid to cell suspension cultures of Vanilla planifolia resulted in the formation of 4-hydroxybenzoic acid, vanillic acid, and syringic acid, respectively. The homologous 4-methoxybenzoic acids were demethylated to the same products. It is concluded that the side chain degrading enzyme system accepts the 4-methoxylated substrates while the demethylation occurs at the benzoic acid level. The demethylating enzyme is specific for the 4-position. Feeding of [O-(14)C-methyl]-3,4-dimethoxycinnamic acid revealed that the first step in the conversion is the glycosylation of the cinnamic acid to its glucose ester. A partial purification of a UDP-glucose: trans-cinnamic acid glucosyltransferase is reported. 4-Methoxy substituted cinnamic acids are better substrates for this enzyme than 4-hydroxy substituted cinnamic acid. It is suggested that 4-methoxy substituted cinnamic acids are intermediates in the biosynthetic conversion of cinnamic acids to benzoic acids in cells of V. planifolia.

  7. Methyl Jasmonate and Salicylic Acid Induced Oxidative Stress and Accumulation of Phenolics in Panax ginseng Bioreactor Root Suspension Cultures

    Directory of Open Access Journals (Sweden)

    Kee-Yoeup Paek

    2007-03-01

    Full Text Available To investigate the enzyme variations responsible for the synthesis of phenolics, 40 day-old adventitious roots of Panax ginseng were treated with 200 μM methyl jasmonate (MJ or salicylic acid (SA in a 5 L bioreactor suspension culture (working volume 4 L. Both treatments caused an increase in the carbonyl and hydrogen peroxide (H2O2 contents, although the levels were lower in SA treated roots. Total phenolic, flavonoid, ascorbic acid, non-protein thiol (NPSH and cysteine contents and 1,1-diphenyl-2-picrylhydrazyl (DPPH radical reducing activity were increased by MJ and SA. Fresh weight (FW and dry weight (DW decreased significantly after 9 days of exposure to SA and MJ. The highest total phenolics (62%, DPPH activity (40%, flavonoids (88%, ascorbic acid (55%, NPSH (33%, and cysteine (62% contents compared to control were obtained after 9 days in SA treated roots. The activities of glucose 6-phosphate dehydrogenase, phenylalanine ammonia lyase, substrate specific peroxidases (caffeic acid peroxidase, quercetin peroxidase and ferulic acid peroxidase were higher in MJ treated roots than the SA treated ones. Increased shikimate dehydrogenase, chlorogenic acid peroxidase and β-glucosidase activities and proline content were observed in SA treated roots than in MJ ones. Cinnamyl alcohol dehydrogenase activity remained unaffected by both MJ and SA. These results strongly indicate that MJ and SA induce the accumulation of phenolic compounds in ginseng root by altering the phenolic synthesis enzymes.

  8. Monitoring the differentiation and migration patterns of neural cells derived from human embryonic stem cells using a microfluidic culture system.

    Science.gov (United States)

    Lee, Nayeon; Park, Jae Woo; Kim, Hyung Joon; Yeon, Ju Hun; Kwon, Jihye; Ko, Jung Jae; Oh, Seung-Hun; Kim, Hyun Sook; Kim, Aeri; Han, Baek Soo; Lee, Sang Chul; Jeon, Noo Li; Song, Jihwan

    2014-06-01

    Microfluidics can provide unique experimental tools to visualize the development of neural structures within a microscale device, which is followed by guidance of neurite growth in the axonal isolation compartment. We utilized microfluidics technology to monitor the differentiation and migration of neural cells derived from human embryonic stem cells (hESCs). We co-cultured hESCs with PA6 stromal cells, and isolated neural rosette-like structures, which subsequently formed neurospheres in suspension culture. Tuj1-positive neural cells, but not nestin-positive neural precursor cells (NPCs), were able to enter the microfluidics grooves (microchannels), suggesting that neural cell-migratory capacity was dependent upon neuronal differentiation stage. We also showed that bundles of axons formed and extended into the microchannels. Taken together, these results demonstrated that microfluidics technology can provide useful tools to study neurite outgrowth and axon guidance of neural cells, which are derived from human embryonic stem cells.

  9. A Novel Hydroxyproline-Deficient Arabinogalactan Protein Secreted by Suspension-Cultured Cells of Daucus carota (Purification and Partial Characterization).

    Science.gov (United States)

    Baldwin, T. C.; McCann, M. C.; Roberts, K.

    1993-09-01

    Arabinogalactan proteins (AGPs) are secreted or membrane-associated glycoproteins that have been operationally defined as binding to [beta]-glucosyl Yariv artificial antigen, being rich in arabinose and galactose, and containing high levels of alanine, serine, and hydroxyproline. Using an anti-AGP monoclonal antibody (MAC 207) bound to cyanogen bromide-activated Sepharose 4B, we have purified by immunoaffinity chromatography an extracellular AGP from the culture medium of suspension-cultured cells of carrot (Daucus carota). The apparent molecular mass of this highly glycosylated proteoglycan is 70 to 100 kD as judged by sodium dodecyl sulfate-polyacrylamide gels. Although its sugar analysis, [beta]-glucosyl Yariv binding, and high alanine, serine, and proline content are consistent with it being an AGP, the amino acid composition unexpectedly revealed this molecule to have no detectable hydroxyproline. This suggests that this glycoprotein is not a "classical" AGP, but represents the first example of a new class of hydroxyproline-poor AGPs. Deglycosylation of the AGP with anhydrous hydrogen fluoride revealed that the purified proteoglycan contains probably a single core protein with an apparent molecular mass of 30 kD. Direct visualization of the native AGP in the electron microscope showed ellipsoidal putative AGP monomers, approximately 25 nm by 15 nm, that showed a strong tendency to self assemble into higher-order structures. Upon desiccation, the glycosylated AGP formed paracrystalline arrays visible in the light microscope. Polarized Fourier transform infrared microspectroscopy of these arrays demonstrated a high degree of polarization of the sugar moieties under these conditions. These results put possible constraints on current models of AGP structure; a putative role for these novel AGPs as pectin-binding proteins is discussed.

  10. Generation of highly productive polyclonal and monoclonal tobacco suspension lines from a heterogeneous transgenic BY-2 culture through flow cytometric sorting

    OpenAIRE

    Kirchhoff, Janina

    2012-01-01

    Over the last 20 years, plant cells gained increasing interest as host systems for the production of human and animal pharmaceuticals. Transgenic tobacco suspension cultures are an especially promising production host because of their beneficial growth, low maintenance requirement and the possibility of contained cultivation in bioreactors under controlled conditions. The excellent product quality of plant-produced proteins was demonstrated for a multitude of proteins, but the heterogeneous a...

  11. Detection of anthraquinones and identification of 1,4-naphthohydroquinone in cell suspension cultures of Rudgea jasminoides (Cham.) Müll. Arg. (Rubiaceae)

    OpenAIRE

    Oliveira, Marisa de Cacia; Negri, Giusepina; Salatino, Antônio; Braga, Márcia Regina

    2007-01-01

    In Rubiaceae, anthraquinones and naphthoquinones are secondary metabolites characteristic of the subfamily Rubioideae, in which Rudgea jasminoides is included. Thin-layer chromatography using specific solvent systems and spray reagents indicated the presence of anthraquinones constitutively produced by cell suspension cultures of R. jasminoides. GC/MS analysis detected 1,4-naphthohydroquinone as a product of biosynthesis only after elicitation of the cells with yeast extract (Saccharomyces ce...

  12. Innovative, non-stirred bioreactors in scales from milliliters up to 1000 liters for suspension cultures of cells using disposable bags and containers--a Swiss contribution.

    Science.gov (United States)

    Werner, Sören; Eibl, Regine; Lettenbauer, Christine; Röll, Marcel; Eibl, Dieter; De Jesus, Maria; Zhang, Xiaowei; Stettler, Matthieu; Tissot, Stephanie; Bürki, Cedric; Broccard, Gilles; Kühner, Markus; Tanner, Rolf; Baldi, Lucia; Hacker, David; Wurm, Florian M

    2010-01-01

    Innovative mixing principles in bioreactors, for example using the rocking of a platform to induce a backwards and forwards 'wave', or using orbital shaking to generate a 'wave' that runs round in a cylindrical container, have proved to be successful for the suspension cultures of cells, especially when combined with disposable materials. This article presents an overview of the engineering characteristics when these new principles are applied in bioreactors, and case studies covering scales of operation from milliliters to 1000 liters.

  13. Cell suspension culture-mediated incorporation of the rice bel gene into transgenic cotton.

    Directory of Open Access Journals (Sweden)

    Liping Ke

    Full Text Available Cotton plants engineered for resistance to the herbicides, glyphosate or glufosinate have made a considerable impact on the production of the crop worldwide. In this work, embryogenic cell cultures derived from Gossypium hirsutum L. cv Coker 312 hypocotyl callus were transformed via Agrobacterium tumefaciens with the rice cytochrome P450 gene, CYP81A6 (bel. In rice, bel has been shown to confer resistance to both bentazon and sulfanylurea herbicides. Transformed cells were selected on a liquid medium supplemented alternately or simultaneously with kanamycin (50mg/L and bentazon (4.2 µmol. A total of 17 transgenic cotton lines were recovered, based on the initial resistance to bentazon and on PCR detection of the bel transgene. Bel integration into the cotton genome was confirmed by Southern blot and expression of the transgene was verified by RT-PCR. In greenhouse and experimental plot trials, herbicide (bentazon tolerance of up to 1250 mg/L was demonstrated in the transgenic plants. Transgenic lines with a single copy of the bel gene showed normal Mendelian inheritance of the characteristic. Importantly, resistance to bentazon was shown to be stably incorporated in the T1, T2 and T3 generations of self-fertilised descendents and in plants outcrossed to another upland cotton cultivar. Engineering resistance to bentazon in cotton through the heterologous expression of bel opens the possibility of incorporating this trait into elite cultivars, a strategy that would give growers a more flexible alternative to weed management in cotton crops.

  14. Fast Filtration of Bacterial or Mammalian Suspension Cell Cultures for Optimal Metabolomics Results.

    Directory of Open Access Journals (Sweden)

    Natalie Bordag

    Full Text Available The metabolome offers real time detection of the adaptive, multi-parametric response of the organisms to environmental changes, pathophysiological stimuli or genetic modifications and thus rationalizes the optimization of cell cultures in bioprocessing. In bioprocessing the measurement of physiological intracellular metabolite levels is imperative for successful applications. However, a sampling method applicable to all cell types with little to no validation effort which simultaneously offers high recovery rates, high metabolite coverage and sufficient removal of extracellular contaminations is still missing. Here, quenching, centrifugation and fast filtration were compared and fast filtration in combination with a stabilizing washing solution was identified as the most promising sampling method. Different influencing factors such as filter type, vacuum pressure, washing solutions were comprehensively tested. The improved fast filtration method (MxP® FastQuench followed by routine lipid/polar extraction delivers a broad metabolite coverage and recovery reflecting well physiological intracellular metabolite levels for different cell types, such as bacteria (Escherichia coli as well as mammalian cells chinese hamster ovary (CHO and mouse myeloma cells (NS0.The proposed MxP® FastQuench allows sampling, i.e. separation of cells from medium with washing and quenching, in less than 30 seconds and is robustly designed to be applicable to all cell types. The washing solution contains the carbon source respectively the 13C-labeled carbon source to avoid nutritional stress during sampling. This method is also compatible with automation which would further reduce sampling times and the variability of metabolite profiling data.

  15. Purification, molecular cloning, and characterization of glutathione S-transferases (GSTs) from pigmented Vitis vinifera L. cell suspension cultures as putative anthocyanin transport proteins

    Science.gov (United States)

    Conn, Simon; Curtin, Chris; Bézier, Annie; Franco, Chris; Zhang, Wei

    2008-01-01

    The ligandin activity of specific glutathione S-transferases (GSTs) is necessary for the transport of anthocyanins from the cytosol to the plant vacuole. Five GSTs were purified from Vitis vinifera L. cv. Gamay Fréaux cell suspension cultures by glutathione affinity chromatography. These proteins underwent Edman sequencing and mass spectrometry fingerprinting, with the resultant fragments aligned with predicted GSTs within public databases. The corresponding coding sequences were cloned, with heterologous expression in Escherichia coli used to confirm GST activity. Transcriptional profiling of these candidate GST genes and key anthocyanin biosynthetic pathway genes (PAL, CHS, DFR, and UFGT) in cell suspensions and grape berries against anthocyanin accumulation demonstrated strong positive correlation with two sequences, VvGST1 and VvGST4, respectively. The ability of VvGST1 and VvGST4 to transport anthocyanins was confirmed in the heterologous maize bronze-2 complementation model, providing further evidence for their function as anthocyanin transport proteins in grape cells. Furthermore, the differential induction of VvGST1 and VvGST4 in suspension cells and grape berries suggests functional differences between these two proteins. Further investigation of these candidate ligandins may identify a mechanism for manipulating anthocyanin accumulation in planta and in vitro suspension cells. PMID:18836188

  16. Factors influencing cucumber (Cucumis sativus L. somatic embryogenesis. I. The crucial role of pH and nitrogen in suspension culture

    Directory of Open Access Journals (Sweden)

    Tadeusz Wróblewski

    2014-01-01

    Full Text Available A method of obtaining and the characteristics of an embryogenic stabilised cucumber (Cucumis sativus L. suspension culture which has many similarities to the carrot model are presented. The Specific Type I cells and proembryogenic mass were present in such a suspension. The maintenance of the proembryogenic stage took place in medium containing 2,4-D as the sole growth regulator, subsequent stages of embryogenesis occurred in hormone-free medium. Embryonic structures were also observed in medium with auxin in the late stages of growth, probably due to the depletion of 2,4-D in the medium during subculture. The choice of the proper inorganic nitrogen sources and the maintenance of correct proportions between them had a significant effect on the formation of these structures. We have shown that the pH of the medium with an embryogenic culture became stabilized regardless of the initial pH value and depended on the medium composition. The inoculum used for the initiation of subsequent subcultures of the stable suspension culture was 1 part tissue to 300 parts medium and was small in comparison to the systems described for the cucumber so far. From 1 ml of basic suspension 7 embryos were obtained on medium without growth regulators 10 days after inoculation, and this amount increased to 21 after 3 weeks. From 3.2% of the somatic embryos it was posible to regenerate plants. The high yield and synchronisation of the process and the development of embryos without passing through callus tissue create the possibility of using this system for molecular investigations and in the technology of somatic seed production.

  17. Jasmonic Acid Effect on the Fatty Acid and Terpenoid Indole Alkaloid Accumulation in Cell Suspension Cultures of Catharanthus roseus

    Directory of Open Access Journals (Sweden)

    Guitele Dalia Goldhaber-Pasillas

    2014-07-01

    Full Text Available The stress response after jasmonic acid (JA treatment was studied in cell suspension cultures of Catharanthus roseus. The effect of JA on the primary and secondary metabolism was based on changes in profiles of fatty acids (FA and terpenoid indole alkaloids (TIA. According to multivariate data analyses (MVDA, three major time events were observed and characterized according to the variations of specific FA and TIA: after 0–30 min of induction FA such as C18:1, C20:0, C22:0 and C24:0 were highly induced by JA; 90–360 min after treatment was characterized by variations of C14:0 and C15:0; and 1440 min after induction JA had the largest effect on both group of metabolites were C18:1, C18:2, C18:3, C16:0, C20:0, C22:0, C24:0, catharanthine, tabersonine-like 1, serpentine, tabersonine and ajmalicine-like had the most significant variations. These results unambiguously demonstrate the profound effect of JA particularly on the accumulation of its own precursor, C18:3 and the accumulation of TIA, which can be considered as late stress response events to JA since they occurred only after 1440 min. These observations show that the early events in the JA response do not involve the de novo biosynthesis of neither its own precursor nor TIA, but is due to an already present biochemical system.

  18. [Effect of vanadium compounds on the growth and production of coumarins in the suspension culture of Angelica archangelica L].

    Science.gov (United States)

    Siatka, T; Kasparová, M

    2007-10-01

    Plant tissue cultures represent a promising source of substances of natural origin. The main problem of their use is a low production of the majority of secondary metabolites. One of the methods of increasing the production of these substances is elicitation, because the biosynthesis of many secondary metabolites in plant cells is part of the defensive reaction against biological or abiotic stress influences. The paper tested vanadium compounds as potential elicitors of the production of coumarins: sodium vanadate (0.2; 1; 10; 100 and 1000 microM/l of medium) and vanadyl sulfate (10; 20; 50; 100; 200 and 500 microM/l of medium). The toxicity of these substances for the culture was simultaneously monitored by means of the evaluation of the effects on growth (characterized by fresh and dry weights of biomass at the end of two-week cultivation). The cultures were grown both in the light and dark. The growth of the cultures was not influenced by sodium vanadate at concentrations of 0.2 to 100 microM. A vanadate concentration of 1000 microM acted already toxically (in comparison with the control culture, the fresh weight was decreased by 28 % and the dry weight by 41% when cultivated in the light; the fresh weight by 69% and the dry weight by 66% when cultivated in the dark). Vanadyl sulfate in concentrations of 10 to 50 microM did not affect the growth of the culture, at higher concentrations it decreased it gradually: a concentration of 500 microM acted already toxically, again more markedly when cultivated in the light (in comparison with the control culture, the fresh weight was decreased by 27% and dry weight by 38% when cultivated in the light; the fresh weight by 65% and the dry weight by 61% when cultivated in the dark). The production of coumarins was stimulated by sodium vanadate in a concentration of 0.2 and 1 microM when cultivated in the light. The content of coumarins increased in comparison with the control culture mainly in the medium by 46% and by 25

  19. Positive selection of Wharton's jelly-derived CD105(+) cells by MACS technique and their subsequent cultivation under suspension culture condition: A simple, versatile culturing method to enhance the multipotentiality of mesenchymal stem cells.

    Science.gov (United States)

    Amiri, Fatemeh; Halabian, Raheleh; Dehgan Harati, Mozhgan; Bahadori, Marzie; Mehdipour, Ahmad; Mohammadi Roushandeh, Amaneh; Habibi Roudkenar, Mehryar

    2015-05-01

    Wharton's jelly (WJ), an appropriate source of mesenchymal stem cells (MSCs), has been shown to have a wide array of therapeutic applications. However, the WJ-derived MSCs are very heterogeneous and have limited expression of pluripotency markers. Hence, improvement of their culture condition would promote the efficiency of WJ-MSCs. This study aims to employ a simple method of cultivation to obtain WJ-MSCs which express more pluripotency markers. CD105(+) cells were separated by magnetic-associated (activated) cell sorting from umbilical cord mucous tissue. CD105(+) cells were added to Methocult medium diluted in α-minimum essential medium (α-MEM) and seeded in poly(2-hydroxyethyl methacrylate) (poly-HEMA)-coated plates for suspension culture preparation. Differentiation capacity of isolated cells was evaluated in the presence of differentiation-inducing media. The expression of pluripotency markers such as Oct3/4, Nanog, and Sox2 was also analyzed by RT-PCR and western blot techniques. Moreover, immunocytochemistry was performed to detect alpha-smooth muscle actin (antigene) (α-SMA) protein. WJ-MSCs grew homogeneously and formed colonies when cultured under suspension culture conditions (Non-adhesive WJ-MSCs). They maintained their growth ability in both adherent and suspension cultures for several passages. Non-adhesive WJ-MSCs expressed Oct3/4, Nanog, and Sox2 both at transcriptional and translational levels in comparison to those cultured in conventional adherent cultures. They also expressed α-SMA protein. In this study, we isolated WJ-MSCs using a slightly modified culture condition. Our simple non-genetic method resulted in a homogeneous population of WJ-MSCs, which highly expressed pluripotency markers. In the future, more multipotent WJ-MSCs can be harnessed as a non-embryonic source of MSCs in MSC-based cell therapy.

  20. Cell Suspension Culture of Eriobotrya japonica Regulates the Diabetic and Hyperlipidemic Signs of High-Fat-Fed Mice

    Directory of Open Access Journals (Sweden)

    Jin-Bin Wu

    2013-03-01

    Full Text Available The present study investigates the anti-hyperlipidemic and antihyperglycemic effects and mechanism in high-fat (HF-fed mice of cell suspension culture of Eriobotrya japonica (TA, which contains a great number of pentacyclic terpenoids. Firstly, C57BL/6J mice were randomly divided into two groups: the control (CON group was fed with a low-fat diet (n = 9, whereas the experimental group was fed a 45% HF diet for 8 weeks. Afterwards, the CON group was treated with vehicle, whereas the HF group was subdivided into five groups and was orally given TA or rosiglitazone or not for 4 weeks. Blood and visceral adipose tissue, liver tissue and skeletal muscle were examined. Treatment with TA reduced body weight gain, weights of white adipose tissue (WAT (including epididymal, perirenal, mesenteric WAT and visceral fat, and hepatic triacylglycerol content significantly without affecting food intake in diet-induced diabetic mice. TA effectively prevented HF diet-induced increases in the levels of blood glucose, insulin, leptin and HOMA-IR index (p < 0.001, p < 0.05, p < 0.05, p < 0.01, respectively and attenuated insulin resistance. Treatment with TA, adipocytes in the visceral depots showed a reduction in size. TA effectively significantly increased the protein contents of phosphorylation of AMPK-α (Thr172 both in liver and adipose tissue. It is shown that TA exhibits hypolipidemic effect in HF-fed mice by decreasing gene expressions of fatty acid synthesis, including acyl-coenzyme A: diacylglycerol acyltransferase (DGAT 2, which catalyzes the final step in the synthesis of triglycerides, and antidiabetic properties occurred as a result of decreased hepatic glucose production via phosphenolpyruvate carboxykinase (PEPCK down- regulation, improved insulin sensitization and TA (at 1.0 g/kg dose decreased expression of hepatic and adipose 11-β-hydroxysteroid dehydroxygenase (11β-HSD1 gene, which contributed in attenuating diabetic state. Futhermore, TA at

  1. Effect of sucrose and methyl jasmonate on biomass and anthocyanin production in cell suspension culture of Melastoma malabathricum (Melastomaceae

    Directory of Open Access Journals (Sweden)

    Koay Suan See

    2011-06-01

    Full Text Available Melastoma malabathricum, belongs to the Melastomaceae family, is an important medicinal plant widely distributed from Madagascar to Australia, that is used in traditional remedies for the treatment of variousailments. Besides its medicinal properties, it has been identified as a potential source of anthocyanin production.The present study was carried out to investigate the effect of sucrose and methyl jasmonate and feeding time oncell biomass yield and anthocyanin production in cell suspension culture of M. malabathricum. Addition of differentconcentrations of sucrose into the cell culture of M. malabathricum influenced cell biomass and pigment accumulation. The addition of methyl jasmonate was found to have no effect on cell biomass but the presence of higher amount (12.5-50mg/L had caused a reduction in anthocyanin production and accumulation. MS medium supplemented with 30g/L sucrose and 3.5 mg/L of MeJA added on cero day and 3rd day produced high fresh cell mass at the end of nine days of culture but did not support the production of anthocyanins. However, cells cultured in the medium supplemented with 45g/L sucrose without MeJA showed the highest pigment content (0.69±0.22Cv/g-FCM. The cells cultured in MS medium supplemented with 30 g/L sucrose with 3.5mg/L MeJA added on the 3rd and 6th day of culture, showed the lowest pigment content (0.37-0.40Cv/g-FCM. This study indicated that MeJA was not necessary but sucrose was needed for the enhancement of cell growth and anthocyanin production in M. malabathricum cell cultures. Rev. Biol. Trop. 59 (2: 597-606. Epub 2011 June 01.elastoma malabathricum pertenece a la familia de las melastomáceas, es una planta medicinal importante ampliamente distribuida desde Madagascar hasta Australia, que se utiliza en remedios tradicionales para el tratamiento de diversas dolencias. Además de sus propiedades medicinales, se ha identificado como una fuente potencial de producción de antocianinas. En esta

  2. Effective Rho-associated protein kinase inhibitor treatment to dissociate human iPS cells for suspension culture to form embryoid body-like cell aggregates.

    Science.gov (United States)

    Horiguchi, Ayumi; Yazaki, Koyuki; Aoyagi, Mami; Ohnuki, Yoshitsugu; Kurosawa, Hiroshi

    2014-11-01

    Treatment conditions using Y-27632 in the preparation of cell suspension of dissociated human pluripotent stem cells (hiPSCs) were investigated in the context of embryoid body (EB)-like cell aggregates. The effectiveness of a pretreatment with Y-27632 before cell dissociation and that of a Y-27632 treatment during cell dissociation were investigated from the viewpoint of simplicity and robustness. The duration of Y-27632 treatment in the preparation process affected the circularity and agglomeration of dissociated hiPSCs. A single application of pretreatment failed to prevent the onset of blebbing. However, a pretreatment promoted the agglomeration of dissociated hiPSCs when combined with the addition of Y-27632 to cell suspension. Our results indicate that pretreatment enhances the agglomeration potential of dissociated hiPSCs. When cell dissociation was performed in the presence of Y-27632, dissociated hiPSCs possessed the highest circularity and significant agglomerating property. It was shown that treatment with Y-27632 during cell dissociation is a simple and robust method to prepare dissociated hiPSCs for suspension culture to form EB-like cell aggregates. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  3. LC-MS determination of steroidal glycosides from Dioscorea deltoidea Wall cell suspension culture: Optimization of pre-LC-MS procedure parameters by Latin Square design.

    Science.gov (United States)

    Sarvin, Boris; Fedorova, Elizaveta; Shpigun, Oleg; Titova, Maria; Nikitin, Mikhail; Kochkin, Dmitry; Rodin, Igor; Stavrianidi, Andrey

    2018-03-30

    In this paper, the ultrasound assisted extraction method for isolation of steroidal glycosides from D. deltoidea plant cell suspension culture with a subsequent HPLC-MS determination was developed. After the organic solvent was selected via a two-factor experiment the optimization via Latin Square 4 × 4 experimental design was carried out for the following parameters: extraction time, organic solvent concentration in extraction solution and the ratio of solvent to sample. It was also shown that the ultrasound assisted extraction method is not suitable for isolation of steroidal glycosides from the D. deltoidea plant material. The results were double-checked using the multiple successive extraction method and refluxing extraction. Optimal conditions for the extraction of steroidal glycosides by the ultrasound assisted extraction method were: extraction time, 60 min; acetonitrile (water) concentration in extraction solution, 50%; the ratio of solvent to sample, 400 mL/g. Also, the developed method was tested on D. deltoidea cell suspension cultures of different terms and conditions of cultivation. The completeness of the extraction was confirmed using the multiple successive extraction method. Copyright © 2018 Elsevier B.V. All rights reserved.

  4. Temperature response of the cell cycle of Haplopappus gracilis in suspension culture and its significance to the G1 transition probability model.

    Science.gov (United States)

    Gould, A R

    1977-01-01

    The effects of temperature on the cell cycle of Haplopappus gracilis suspension cultures were analysed by the fraction of labelled mitoses method. Sphase in these cultures shows a different temperature optimum as compared to optima derived for G2 and mitosis. G1 phase has a much lower Q10 than the other cell cycle phases and shows no temperature optimum between 22 and 34° C. These results are discussed in relation to a transition probability model of the cell cycle proposed by Smith and Martin (Proc. Natl. Acad. Sci. USA 70, 1263-1267, 1973), in which each cell has a time independent probability of initiating the transition into another round of DNA replication and division. The implications of such a model for cell cycle analysis are discussed and a tentative model for a probabilistic transition trigger is advanced.

  5. Use of embryogenic cell suspension and meristem-tip cultures for mutation breeding of apomictic Musa species

    International Nuclear Information System (INIS)

    Novak, F.J.; Afza, R.; Duren, M. van

    1990-01-01

    Full text: Breeding by crossing is difficult for banana and plantain. The plants are heterozygous, therefore mutagenic treatment may uncover a recessive allele by mutating or deleting a corresponding dominant allele. Meristem tips were excised from in vitro growing shoots and used for mutation experiments. Induction was carried out by irradiating shoot tips with γ rays and/or by treatment of explants with ethylmethanesulfonate (EMS). Cell suspension was initiated from corm and leaf tissue excised from in vitro grown plantlets. Mutagenised cell suspensions were derived from leaf and corm tissues irradiated with 60 Co γ rays - (10 to 60 Gy, 8 Gy/min). Musa clones exhibited differences in radiosensitivity and post-irradiation recovery. Doses of 20 to 40 Gy seem suitable for mutation induction. The EMS concentration of 25 mM for 4 hours was found effective for isolated shoot tips. Considerable phenotypic variation was observed among plants regenerated from in vitro shoot tips after mutagenic treatment. Leaf and corm explants kept their morphogenic ability in embryogenic cell suspensions after irradiation up to 25 Gy. (author)

  6. Botulinum hemagglutinin-mediated in situ break-up of human induced pluripotent stem cell aggregates for high-density suspension culture.

    Science.gov (United States)

    Nath, Suman C; Tokura, Tomohiro; Kim, Mee-Hae; Kino-Oka, Masahiro

    2018-04-01

    Large numbers of human induced pluripotent stem cells (hiPSCs) are required for making stable cell bank. Although suspension culture yields high cell numbers, there remain unresolved challenges for obtaining high-density of hiPSCs because large size aggregates exhibit low growth rates. Here, we established a simple method for hiPSC aggregate break-up using botulinum hemagglutinin (HA), which specifically bound with E-cadherin and disrupted cell-cell connections in hiPSC aggregates. HA showed temporary activity for disrupting the E-cadherin-mediated cell-cell connections to facilitate the break-up of aggregates into small sizes only 9 hr after HA addition. The transportation of HA into the aggregates was mediated by transcellular and paracellular way after HA addition to the culture medium. hiPSC aggregates broken up by HA showed a higher number of live cells, higher cell density, and higher expansion fold compared to those of aggregates dissociated with enzymatic digestion. Moreover, a maximum cell density of 4.5 ± 0.2 × 10 6 cells ml -1 was obtained by aggregate break-up into small ones, which was three times higher than that with the conventional culture without aggregate break-up. Therefore, the temporary activity of HA for disrupting E-cadherin-mediated cell-cell connection was key to establishing a simple in situ method for hiPSC aggregate break-up in bioreactors, leading to high cell density in suspension culture. © 2017 Wiley Periodicals, Inc.

  7. Unexpected features of exponentially growing Tobacco Bright Yellow-2 cell suspension culture in relation to excreted extracellular polysaccharides and cell wall composition.

    Science.gov (United States)

    Issawi, Mohammad; Muhieddine, Mohammad; Girard, Celine; Sol, Vincent; Riou, Catherine

    2017-10-01

    This article presents a new insight about TBY-2 cells; from extracellular polysaccharides secretion to cell wall composition during cell suspension culture. In the medium of cells taken 2 days after dilution (end of lag phase), a two unit pH decrease from 5.38 to 3.45 was observed and linked to a high uronic acid (UA) amount secretion (47.8%) while, in 4 and 7 day-old spent media, pH increased and UA amounts decreased 35.6 and 42.3% UA, respectively. To attain deeper knowledge of the putative link between extracellular polysaccharide excretion and cell wall composition, we determined cell wall UA and neutral sugar composition of cells from D2 to D12 cultures. While cell walls from D2 and D3 cells contained a large amount of uronic acid (twice as much as the other analysed cell walls), similar amounts of neutral sugar were detected in cells from lag to end of exponential phase cells suggesting an enriched pectin network in young cultures. Indeed, monosaccharide composition analysis leads to an estimated percentage of pectins of 56% for D3 cell wall against 45% D7 cell walls indicating that the cells at the mid-exponential growth phase re-organized their cell wall linked to a decrease in secreted UA that finally led to a stabilization of the spent medium pH to 5.4. In conclusion, TBY-2 cell suspension from lag to stationary phase showed cell wall remodeling that could be of interest in drug interaction and internalization study.

  8. Comparative study of withanolide production and the related transcriptional responses of biosynthetic genes in fungi elicited cell suspension culture of Withania somnifera in shake flask and bioreactor.

    Science.gov (United States)

    Ahlawat, Seema; Saxena, Parul; Ali, Athar; Khan, Shazia; Abdin, Malik Z

    2017-05-01

    Ashwagandha (Withania somnifera) is one of the most reputed medicinal plants in the traditional medicinal system. In this study, cell suspension culture of W. somnifera was elicited with cell homogenates of fungi (A. alternata, F. solani, V. dahliae and P. indica) in shake flask and the major withanolides like withanolide A, withaferin A and withanone were analysed. Simultaneously expression levels of key pathway genes from withanolides biosynthetic pathways were also checked via quantitative PCR in shake flask as well as in bioreactor. The results show that highest gene expression of 10.8, 5.8, 4.9, and 3.3 folds were observed with HMGR among all the expressed genes in cell suspension cultures with cell homogenates of 3% P. indica, 5% V. dahliae, 3% A. alternata and 3% F. solani, respectively, in comparison to the control in shake flask. Optimized concentration of cell homogenate of P. indica (3% v/v) was added to the growing culture in 5.0-l bioreactor under optimized up-scaling conditions and harvested after 22 days. The genes of MVA, MEP and withanolides biosynthetic pathways like HMGR, SS, SE, CAS, FPPS, DXR and DXS were up-regulated by 12.5, 4.9, 2.18, 4.65, 2.34, 1.89 and 1.4 folds, respectively in bioreactor. The enhancement of biomass (1.13 fold) and withanolides [withanolide A (1.7), withaferin A (1.5), and withanone (1.5) folds] in bioreactor in comparison to shake flask was also found to be in line with the up-regulation of genes of withanolide biosynthetic pathways. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  9. Effects of zinc and cadmium ions on cell growth and production of coumarins in cell suspension cultures of Angelica archangelica L.

    Science.gov (United States)

    Siatka, Tomáš; Kašparová, Marie; Spilková, Jiřina

    2012-12-01

    The plant cell may respond to the excess of heavy metals in its environment by various mechanisms, including enhanced biosynthesis of secondary metabolites. In this study, zinc (0 to 1500 μM) and cadmium ions (0 to 100 μM) were tested as potential elicitors of the production of coumarins in angelica cell suspension cultures. In addition, the toxicity of both metals was assessed by evaluating their effect on cell growth (characterized by fresh and dry biomass at the end of a two-week subculture). It has been found that fresh biomass was not influenced up to zinc concentrations of 150 and 300 μM in the dark-grown and light-grown cultures, resp. Then it declined with an increasing zinc level. Zinc at 1500 μM diminished it by 54% and 24% in the dark-grown and light-grown cultures, resp. Dry biomass was influenced in a similar way. Zinc at 1500 μM reduced dry cell weight by 30% and 20% in cultures in the dark and in the light, resp. Cadmium ions did not affect fresh and dry weights of cells up to concentrations of 10 μM and 50 μM in cultures in the dark and in the light, resp. Toxic concentrations of cadmium are by an order of magnitude lower than those of zinc. Cadmium at 50 μM reduced fresh and dry cell weights by 66% and 59%, resp., in the dark-grown cultures. Cadmium at 100 μM caused a decrease in fresh and dry biomass by 40% and 44%, resp., in the light-grown cultures. Neither zinc nor cadmium improved production of coumarins.

  10. The age-dependent epigenetic and physiological changes in an Arabidopsis T87 cell suspension culture during long-term cultivation

    International Nuclear Information System (INIS)

    Kwiatkowska, Aleksandra; Zebrowski, Jacek; Oklejewicz, Bernadetta; Czarnik, Justyna; Halibart-Puzio, Joanna; Wnuk, Maciej

    2014-01-01

    Highlights: • A decrease in proliferation rate during long-term cultivation of Arabidopsis cells. • Age-dependent increase in senescence-associated gene expression in Arabidopsis cells. • Age-related increase in DNA methylation, H3K9me2, and H3K27me3 in Arabidopsis cells. • High potential of photosynthetic efficiency of long-term cultured Arabidopsis cells. - Abstract: Plant cell suspension cultures represent good model systems applicable for both basic research and biotechnological purposes. Nevertheless, it is widely known that a prolonged in vitro cultivation of plant cells is associated with genetic and epigenetic instabilities, which may limit the usefulness of plant lines. In this study, the age-dependent epigenetic and physiological changes in an asynchronous Arabidopsis T87 cell culture were examined. A prolonged cultivation period was found to be correlated with a decrease in the proliferation rate and a simultaneous increase in the expression of senescence-associated genes, indicating that the aging process started at the late growth phase of the culture. In addition, increases in the heterochromatin-specific epigenetic markers, i.e., global DNA methylation, H3K9 dimethylation, and H3K27 trimethylation, were observed, suggesting the onset of chromatin condensation, a hallmark of the early stages of plant senescence. Although the number of live cells decreased with an increase in the age of the culture, the remaining viable cells retained a high potential to efficiently perform photosynthesis and did not exhibit any symptoms of photosystem II damage

  11. Suspension culture process of MethA tumor cell for the production of heat-shock protein glycoprotein 96: process optimization in spinner flasks.

    Science.gov (United States)

    Tang, Ya-Jie; Li, Hong-Mei; Hamel, Jean-François P

    2007-01-01

    Heat-shock proteins (HSPs) act like "chaperones", making sure that the cell's proteins are in the right shape and in the right place at the right time. Heat-shock protein glycoprotein 96 (gp96) is a member of the HSP90 protein family, which chaperones a number of molecules in protein folding and transportation. Heat-shock protein gp96 serves as a natural adjuvant for chaperoning antigenic peptides into the immune surveillance pathways. Currently, heat-shock protein gp96 was only isolated from murine and human tissues and cell lines. An animal cell suspension culture process for the production of heat-shock protein gp96 by MethA tumor cell was developed for the first time in spinner flasks. Effects of culture medium and condition were studied to enhance the MethA tumor cell density and the production and productivity of heat-shock protein gp96. Initial glucose concentration had a significant effect on the heat-shock protein gp96 accumulation, and an initial glucose level of 7.0 g/L was desirable for MethA tumor cell growth and heat-shock protein gp96 production and productivity. Cultures at an initial glutamine concentration of 3 and 6 mM were nutritionally limited by glutamine. At an initial glutamine concentration of 6 mM, the maximal viable cell density of 19.90 x 10(5) cells/mL and the maximal heat-shock protein gp96 production of 4.95 mg/L was obtained. The initial concentration of RPMI 1640 and serum greatly affected the MethA tumor cell culture process. Specifically cultures with lower initial concentration of RPMI 1640 resulted in lower viable cell density and lower heat-shock protein gp96 production. At an initial serum concentration of 8%, the maximal viable cell density of 19.18 x 10(5) cells/mL and the maximal heat-shock protein gp96 production of 5.67 mg/L was obtained. The spin rate significantly affected the cell culture process in spinner flasks, and a spin rate of 150 rpm was desirable for MethA tumor cell growth and heat-shock protein gp96

  12. The Metabolic Fate of Deoxynivalenol and Its Acetylated Derivatives in a Wheat Suspension Culture: Identification and Detection of DON-15-O-Glucoside, 15-Acetyl-DON-3-O-Glucoside and 15-Acetyl-DON-3-Sulfate

    Directory of Open Access Journals (Sweden)

    Clemens Schmeitzl

    2015-08-01

    Full Text Available Deoxynivalenol (DON is a protein synthesis inhibitor produced by the Fusarium species, which frequently contaminates grains used for human or animal consumption. We treated a wheat suspension culture with DON or one of its acetylated derivatives, 3-acetyl-DON (3-ADON, 15-acetyl-DON (15-ADON and 3,15-diacetyl-DON (3,15-diADON, and monitored the metabolization over a course of 96 h. Supernatant and cell extract samples were analyzed using a tailored LC-MS/MS method for the quantification of DON metabolites. We report the formation of tentatively identified DON-15-O-β-D-glucoside (D15G and of 15-acetyl-DON-3-sulfate (15-ADON3S as novel deoxynivalenol metabolites in wheat. Furthermore, we found that the recently identified 15-acetyl-DON-3-O-β-D-glucoside (15-ADON3G is the major metabolite produced after 15-ADON challenge. 3-ADON treatment led to a higher intracellular content of toxic metabolites after six hours compared to all other treatments. 3-ADON was exclusively metabolized into DON before phase II reactions occurred. In contrast, we found that 15-ADON was directly converted into 15-ADON3G and 15-ADON3S in addition to metabolization into deoxynivalenol-3-O-β-D-glucoside (D3G. This study highlights significant differences in the metabolization of DON and its acetylated derivatives.

  13. Production of high-titer human influenza A virus with adherent and suspension MDCK cells cultured in a single-use hollow fiber bioreactor.

    Science.gov (United States)

    Tapia, Felipe; Vogel, Thomas; Genzel, Yvonne; Behrendt, Ilona; Hirschel, Mark; Gangemi, J David; Reichl, Udo

    2014-02-12

    Hollow fiber bioreactors (HFBRs) have been widely described as capable of supporting the production of highly concentrated monoclonal antibodies and recombinant proteins. Only recently HFBRs have been proposed as new single-use platforms for production of high-titer influenza A virus. These bioreactors contain multiple hollow fiber capillary tubes that separate the bioreactor in an intra- and an extra-capillary space. Cells are usually cultured in the extra-capillary space and can grow to a very high cell concentration. This work describes the evaluation of the single-use hollow fiber bioreactor PRIMER HF (Biovest International Inc., USA) for production of influenza A virus. The process was setup, characterized and optimized by running a total of 15 cultivations. The HFBRs were seeded with either adherent or suspension MDCK cells, and infected with influenza virus A/PR/8/34 (H1N1), and the pandemic strain A/Mexico/4108/2009 (H1N1). High HA titers and TCID₅₀ of up to 3.87 log₁₀(HA units/100 μL) and 1.8 × 10(10)virions/mL, respectively, were obtained for A/PR/8/34 influenza strain. Influenza virus was collected by performing multiple harvests of the extra-capillary space during a virus production time of up to 12 days. Cell-specific virus yields between 2,000 and 8,000 virions/cell were estimated for adherent MDCK cells, and between 11,000 and 19,000 virions/cell for suspension MDCK.SUS2 cells. These results do not only coincide with the cell-specific virus yields obtained with cultivations in stirred tank bioreactors and other high cell density systems, but also demonstrate that HFBRs are promising and competitive single-use platforms that can be considered for commercial production of influenza virus. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Spontaneous pepsin C-catalyzed activation of human pepsinogen C in transgenic rice cell suspension culture: Production and characterization of human pepsin C.

    Science.gov (United States)

    Islam, Md Reyazul; Kim, Nan-Sun; Jung, Jae-Wan; Kim, Hyo-Boon; Han, So-Chon; Yang, Moon-Sik

    2018-01-01

    A human pepsinogen C (hPGC) gene was synthesized with rice-optimized codon usage and cloned into a rice expression vector containing the promoter, signal peptide, and terminator derived from the rice α-amylase 3D (Ramy3D) gene. In addition, a 6-His tag was added to the 3' end of the synthetic hPGC gene for easy purification. The plant expression vector was introduced into rice calli (Oryza sativa L. cv. Dongjin) mediated by Agrobacterium tumefaciens. The integration of the hPGC gene into the chromosome of the transgenic rice callus and hPGC expression in transgenic rice cell suspensions was verified via genomic DNA polymerase chain reaction amplification and Northern blot analysis. Western blot analysis indicated both hPGC and its mature form, human pepsin C, with masses of 42- and 36-kDa in the culture medium under sugar starvation conditions. Human pepsin C was purified from the culture medium using a Ni-NTA agarose column and the NH 2 -terminal 5-residue sequences were verified by amino acid sequencing. The hydrolyzing activity of human pepsin C was confirmed using bovine hemoglobin as a substrate. The optimum pH and temperature for pepsin activity were 2.0 and 40°C, respectively. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Wireless Sensor Platform for Cultural Heritage Monitoring and Modeling System

    Science.gov (United States)

    Bermudez, Sergio A.; Schrott, Alejandro G.; Tsukada, Masahiko; Kargere, Lucretia; Marianno, Fernando; Hamann, Hendrik F.; López, Vanessa; Leona, Marco

    2017-01-01

    Results from three years of continuous monitoring of environmental conditions using a wireless sensor platform installed at The Cloisters, the medieval branch of the New York Metropolitan Museum of Art, are presented. The platform comprises more than 200 sensors that were distributed in five galleries to assess temperature and air flow and to quantify microclimate changes using physics-based and statistical models. The wireless sensor network data shows a very stable environment within the galleries, while the dense monitoring enables localized monitoring of subtle changes in air quality trends and impact of visitors on the microclimate conditions. The high spatial and temporal resolution data serves as a baseline study to understand the impact of visitors and building operations on the long-term preservation of art objects. PMID:28858223

  16. Wireless Sensor Platform for Cultural Heritage Monitoring and Modeling System.

    Science.gov (United States)

    Klein, Levente J; Bermudez, Sergio A; Schrott, Alejandro G; Tsukada, Masahiko; Dionisi-Vici, Paolo; Kargere, Lucretia; Marianno, Fernando; Hamann, Hendrik F; López, Vanessa; Leona, Marco

    2017-08-31

    Results from three years of continuous monitoring of environmental conditions using a wireless sensor platform installed at The Cloisters, the medieval branch of the New York Metropolitan Museum of Art, are presented. The platform comprises more than 200 sensors that were distributed in five galleries to assess temperature and air flow and to quantify microclimate changes using physics-based and statistical models. The wireless sensor network data shows a very stable environment within the galleries, while the dense monitoring enables localized monitoring of subtle changes in air quality trends and impact of visitors on the microclimate conditions. The high spatial and temporal resolution data serves as a baseline study to understand the impact of visitors and building operations on the long-term preservation of art objects.

  17. Privacy Issues of Electronic Monitoring Of Employees: A Cross-Cultural Examination Of Gender Differences

    OpenAIRE

    Raymond E. Taylor

    2013-01-01

    This manuscript presents the results of a study which examined the privacy issues of electronic monitoring of employees from a cross-cultural perspective comparing participants from Taiwan with those from the United States. The results of the study suggest that gender differences exist between Taiwanese and American respondents’ attitudes concerning privacy issues of electronic monitoring of employees. The study suggests that monitoring with notice was an important parameter in determining ho...

  18. [The perfusate culture--bacteriologic monitoring of kidney grafts].

    Science.gov (United States)

    Buchholz, B; Zastrow, F; Lison, A E; Ritzerfeld, W

    1985-01-01

    Since the kidney recipient's immune system is entirely suppressed, any bacterial contamination from a graft might be hazardous. Major statistics [1,3,4,5] reveal a mortality as high as 10% due to infectious and gastrointestinal complications. From July 1979 to December 1983 114 kidney grafts have been done in our center. After transplantation none of the patients died as a result of complications due to infection. Microbiologic examination of the perfusate is obligatory to detect contamination. It was used in 145 donor nephrectomies; 28% of the perfusate culture samples were positive: In 4 of 5 cases (81%) the bacteria isolated were of the non-pathogenic type seen in the normal flora of the skin (Staphylococcus epidermidis). Introduction of cover drapes lowered the positive culture rate to 8%. Isolation of S. epidermidis after desinfection of the skin (6x) with 70% spore-free alcohol is proof of the extraordinary sensitivity of the method used. The outstanding clinical importance of this method is the rapid information obtained on any contamination and the early suggestion concerning the first choice of antibiotic. Though E.coli and Pseudomonas aeruginosa were found in the culture, no clinical infection was seen under adequate antimicrobial therapy. Among 114 kidney transplantations in our center no patient died of bacterial infection. Our experience points out that the effect of general antibiotic prophylaxis is negligible. Instead, the effect of early application of antibiotics in accordance with the results of the perfusate culture is superior.

  19. Uptake and metabolism of clomazone in tolerant-soybean and susceptible-cotton photomixotrophic cell suspension cultures

    Energy Technology Data Exchange (ETDEWEB)

    Norman, M.A.; Liebl, R.A.; Widholm, J.M. (Univ. of Illinois, Urbana (USA))

    1990-03-01

    Studies were conducted to determine the uptake and metabolism of the pigment synthesis inhibiting herbicide clomazone in tolerant-soybean (Glycine max (L.) Merr. cv Corsoy) and susceptible-cotton (Gossypium hirsutum (L.) cv Stoneville 825) photomixotrophic cell suspensions. Soybean and cotton on a whole plant level are tolerant and susceptible to clomazone, respectively. Preliminary studies indicated that I{sub 50} values for growth, chlorophyll (Chl), {beta}-carotene, and lutein were, respectively, >22, 14, 19, and 23 times greater for the soybean cell line (SB-M) 8 days after treatment (DAT) compared to the cotton cell line (COT-M) 16 DAT. Differences in ({sup 14}C)clomazone uptake cannot account for selectivity since there were significantly greater levels of domazone absorbed by the SB-M cells compared to the COT-M cells for each treatment. The percentage of absorbed clomazone converted to more polar metabolite(s) was significantly greater by the SB-M cells relative to COT-M cells at 6 and 24 hours after treatment, however, only small differences existed between the cell lines by 48 hours after treatment. Nearly identical levels of parental clomazone was recovered from both cell lines for all treatments. A pooled metabolite fraction isolated from SB-M cells had no effect on the leaf pigment content of susceptible velvetleaf or soybean seedlings. Conversely, a pooled metabolite fraction from COT-M cells reduced the leaf Chl content of velvetleaf. Soybean tolerance to clomazone appears to be due to differential metabolism (bioactivation) and/or differences at the site of action.

  20. Role of cellular antioxidants (glutathione and ascorbic acid) in the growth and development of wild carrot suspension cultures

    International Nuclear Information System (INIS)

    Earnshaw, B.A.

    1986-01-01

    Determinations of endogenous glutathione (GSH), glutathione disulfide (GSSG), ascorbic acid (AA) and dehydroascorbic acid (DHA) in proliferating and developing wild carrot cultures showed that lower levels of GSH and AA were associated with developing cultures. The GSSG and DHA levels did not account for the changes in the levels of antioxidants between proliferating and developing cultures. Studies were designed to test an observed auxin (2,4-Dichlorophenoxyacetic acid, 2,4-D)-antioxidant association. Two fractions (embryo and less developed) were obtained by screening developed cultures which were previously grown in the presence of 14 C-2, 4-D. The embryo fraction had a lower concentration of 14 C than the less developed fraction, supporting the association, since the two fractions showed this relationship with respect to GSH and AA concentrations. Determinations of GSH and AA levels of cells grown in various concentrations of 2,4-D showed the association, decreases in the 2,4-D concentration correlated with decreases in the GSH and AA concentrations. The existence of a respiratory pathway involving GSSG reductase, DHA reductase, and AA oxidase was investigated to test whether inhibition of AA oxidase by 2,4-D could explain the auxin-antioxidant association; however, AA oxidase activity was not detected

  1. Acoustic Methods to Monitor Protein Crystallization and to Detect Protein Crystals in Suspensions of Agarose and Lipidic Cubic Phase

    Energy Technology Data Exchange (ETDEWEB)

    Ericson, Daniel L.; Yin, Xingyu; Scalia, Alexander; Samara, Yasmin N.; Stearns, Richard; Vlahos, Harry; Ellson, Richard; Sweet, Robert M.; Soares, Alexei S.

    2016-02-01

    Improvements needed for automated crystallography include crystal detection and crystal harvesting. A technique that uses acoustic droplet ejection to harvest crystals was previously reported. Here a method is described for using the same acoustic instrument to detect protein crystals and to monitor crystal growth. Acoustic pulses were used to monitor the progress of crystallization trials and to detect the presence and location of protein crystals. Crystals were detected, and crystallization was monitored in aqueous solutions and in lipidic cubic phase. Using a commercially available acoustic instrument, crystals measuring ~150 µm or larger were readily detected. Simple laboratory techniques were used to increase the sensitivity to 50 µm by suspending the crystals away from the plastic surface of the crystallization plate. This increased the sensitivity by separating the strong signal generated by the plate bottom that can mask the signal from small protein crystals. It is possible to further boost the acoustic reflection from small crystals by reducing the wavelength of the incident sound pulse, but our current instrumentation does not allow this option. In the future, commercially available sound-emitting transducers with a characteristic frequency near 300 MHz should detect and monitor the growth of individual 3 µm crystals.

  2. Acoustic Methods to Monitor Protein Crystallization and to Detect Protein Crystals in Suspensions of Agarose and Lipidic Cubic Phase.

    Science.gov (United States)

    Ericson, Daniel L; Yin, Xingyu; Scalia, Alexander; Samara, Yasmin N; Stearns, Richard; Vlahos, Harry; Ellson, Richard; Sweet, Robert M; Soares, Alexei S

    2016-02-01

    Improvements needed for automated crystallography include crystal detection and crystal harvesting. A technique that uses acoustic droplet ejection to harvest crystals was previously reported. Here a method is described for using the same acoustic instrument to detect protein crystals and to monitor crystal growth. Acoustic pulses were used to monitor the progress of crystallization trials and to detect the presence and location of protein crystals. Crystals were detected, and crystallization was monitored in aqueous solutions and in lipidic cubic phase. Using a commercially available acoustic instrument, crystals measuring ~150 µm or larger were readily detected. Simple laboratory techniques were used to increase the sensitivity to 50 µm by suspending the crystals away from the plastic surface of the crystallization plate. This increased the sensitivity by separating the strong signal generated by the plate bottom that can mask the signal from small protein crystals. It is possible to further boost the acoustic reflection from small crystals by reducing the wavelength of the incident sound pulse, but our current instrumentation does not allow this option. In the future, commercially available sound-emitting transducers with a characteristic frequency near 300 MHz should detect and monitor the growth of individual 3 µm crystals. © 2015 Society for Laboratory Automation and Screening.

  3. The Influence of Organisational Culture on Mainstreaming Monitoring and Evaluation in Public Entities Revenue Services

    Directory of Open Access Journals (Sweden)

    K. Sebedi

    2012-12-01

    Full Text Available Research informs us that most of the problems confronting governments can be traced to their inability and failure to appropriately analyze and evaluate organizational culture. This article argues that there is a direct link between organizational culture, organizational performance, as well as monitoring and evaluation. Understanding the effect that organizational culture has, is a significant factor of good governance. The importance of a strong organizational culture together with effective monitoring and evaluation processes contribute appreciably towards efficient performance and effective service delivery. Yet mainstreaming monitoring and evaluation in government departments and entities is often met with varying challenges. Such challenges can be better understood and addressed through aligning the entities’ organizational culture to government’s monitoring and evaluation initiatives. The focus of the article is on responding to the pertinent question: what are the current challenges relative to implementation and how should they be addressed? The article will thus highlight some of the key challenges that are associated with organizational culture, which have direct impact on the mainstreaming of monitoring and evaluation.

  4. Effect of light wavelength on cell growth, content of phenolic compounds and antioxidant activity in cell suspension cultures of Thevetia peruviana.

    Science.gov (United States)

    Arias, J P; Zapata, K; Rojano, B; Arias, M

    2016-10-01

    Thevetia peruviana (T. peruviana) has been considered as a potentially important plant for industrial and pharmacological application. Among the number of compounds which are produced by T. peruviana, antioxidants and polyphenols are of particular interest due to their benefits on human health. Cell suspension cultures of T. peruviana were established under different conditions: 1) constant illumination (24h/day) at different light wavelengths (red, green, blue, yellow and white), 2) darkness and 3) control (12h/12h: day light/dark) to investigate their biomass, substrate uptake, polyphenols production and oxidizing activity. The results showed biomass concentrations between 17.1g dry weight (DW)/l (green light) and 18.2g DW/l (control) after 13days. The cultures that grew under green light conditions consumed completely all substrates after 10days, while other cultures required at least 13days or more. The total phenolic content was between 7.21 and 9.46mg gallic acid (GA)/g DW for all light conditions. In addition the ferric reducing antioxidant power and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid antioxidant activity ranged from 5.41-6.58mg ascorbic acid (AA)/g DW and 82.93-110.39μmol Trolox/g DW, respectively. Interestingly, the samples which grew under the darkness presented a higher phenolic content and antioxidant capacity when compared to the light conditions. All together, these results demonstrate the extraordinary effect of different lighting conditions on polyphenols production and antioxidant compounds by T. peruviana. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Reprogramming of enteroendocrine K cells to pancreatic β-cells through the combined expression of Nkx6.1 and Neurogenin3, and reaggregation in suspension culture

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Esder; Ryu, Gyeong Ryul; Moon, Sung-Dae; Ko, Seung-Hyun; Ahn, Yu-Bae; Song, Ki-Ho, E-mail: kihos@catholic.ac.kr

    2014-01-17

    Highlights: •K cells were selected from STC-1 cells, a heterogeneous enteroendocrine cell line. •K cells did not express Nkx6.1 and Neurogenin3. •Combined expression of Nkx6.1 and Neurogenin3 reprogrammed K cells to β-cells. •Reprogramming of K cells to β-cells was not complete. -- Abstract: Recent studies have demonstrated that adult cells such as pancreatic exocrine cells can be converted to pancreatic β-cells in a process called cell reprogramming. Enteroendocrine cells and β-cells share similar pathways of differentiation during embryonic development. Notably, enteroendocrine K cells express many of the key proteins found in β-cells. Thus, K cells could be reprogrammed to β-cells under certain conditions. However, there is no clear evidence on whether these cells convert to β-cells. K cells were selected from STC-1 cells, an enteroendocrine cell line expressing multiple hormones. K cells were found to express many genes of transcription factors crucial for islet development and differentiation except for Nkx6.1 and Neurogenin3. A K cell clone stably expressing Nkx6.1 (Nkx6.1{sup +}-K cells) was established. Induction of Neurogenin3 expression in Nkx6.1{sup +}-K cells, by either treatment with a γ-secretase inhibitor or infection with a recombinant adenovirus expressing Neurogenin3, led to a significant increase in Insulin1 mRNA expression. After infection with the adenovirus expressing Neurogenin3 and reaggregation in suspension culture, about 50% of Nkx6.1{sup +}-K cells expressed insulin as determined by immunostaining. The intracellular insulin content was increased markedly. Electron microscopy revealed the presence of insulin granules. However, glucose-stimulated insulin secretion was defective, and there was no glucose lowering effect after transplantation of these cells in diabetic mice. In conclusion, we demonstrated that K cells could be reprogrammed partially to β-cells through the combined expression of Nkx6.1 and Neurogenin3, and

  6. Reprogramming of enteroendocrine K cells to pancreatic β-cells through the combined expression of Nkx6.1 and Neurogenin3, and reaggregation in suspension culture

    International Nuclear Information System (INIS)

    Lee, Esder; Ryu, Gyeong Ryul; Moon, Sung-Dae; Ko, Seung-Hyun; Ahn, Yu-Bae; Song, Ki-Ho

    2014-01-01

    Highlights: •K cells were selected from STC-1 cells, a heterogeneous enteroendocrine cell line. •K cells did not express Nkx6.1 and Neurogenin3. •Combined expression of Nkx6.1 and Neurogenin3 reprogrammed K cells to β-cells. •Reprogramming of K cells to β-cells was not complete. -- Abstract: Recent studies have demonstrated that adult cells such as pancreatic exocrine cells can be converted to pancreatic β-cells in a process called cell reprogramming. Enteroendocrine cells and β-cells share similar pathways of differentiation during embryonic development. Notably, enteroendocrine K cells express many of the key proteins found in β-cells. Thus, K cells could be reprogrammed to β-cells under certain conditions. However, there is no clear evidence on whether these cells convert to β-cells. K cells were selected from STC-1 cells, an enteroendocrine cell line expressing multiple hormones. K cells were found to express many genes of transcription factors crucial for islet development and differentiation except for Nkx6.1 and Neurogenin3. A K cell clone stably expressing Nkx6.1 (Nkx6.1 + -K cells) was established. Induction of Neurogenin3 expression in Nkx6.1 + -K cells, by either treatment with a γ-secretase inhibitor or infection with a recombinant adenovirus expressing Neurogenin3, led to a significant increase in Insulin1 mRNA expression. After infection with the adenovirus expressing Neurogenin3 and reaggregation in suspension culture, about 50% of Nkx6.1 + -K cells expressed insulin as determined by immunostaining. The intracellular insulin content was increased markedly. Electron microscopy revealed the presence of insulin granules. However, glucose-stimulated insulin secretion was defective, and there was no glucose lowering effect after transplantation of these cells in diabetic mice. In conclusion, we demonstrated that K cells could be reprogrammed partially to β-cells through the combined expression of Nkx6.1 and Neurogenin3, and reaggregation

  7. Effect of NaCl on ionic content and distribution in suspension-cultured cells of the halophyte Sonneratia alba versus the glycophyte Oryza sativa.

    Science.gov (United States)

    Hayatsu, Manabu; Suzuki, Suechika; Hasegawa, Ai; Tsuchiya, Shinpei; Sasamoto, Hamako

    2014-09-15

    The effect of a high concentration of NaCl on the intra- (cytoplasmic matrix and vacuole) and extracellular (cell wall) distribution of Na, Cl, K, Mg, Ca, S, and P was investigated in suspension-cultured cells of the mangrove halophyte Sonneratia alba and compared to cultured cells of glycophytic rice (Oryza sativa). No significant differences were observed in ultrastructural features of cluster cells of both species cultured with and without 50mM NaCl. Quantitative X-ray microanalysis of cryosections of the cells cultured in the presence of 50mM NaCl showed that the Na concentration ([Na]) and Cl concentration ([Cl]) significantly increased in all three cell components measured. In S. alba, the [Na] was highest in the vacuole and lowest in the cytoplasmic matrix, while the [Cl] was highest in the cell wall and lowest in the cytoplasmic matrix. In O. sativa, however, the [Na] and [Cl] were highest in the cell wall, and the [Na] was lowest in the cytoplasmic matrix. Thus, the possible activities for Na and Cl transport from the cytoplasmic matrix into the vacuole were greater in S. alba than in O. sativa, suggesting that halophilic mangrove cells gain salt tolerance by transporting Na and Cl into their vacuoles. In O. sativa, the addition of NaCl to the culture medium caused no significant changes to the intracellular concentrations of various elements, such as K, P, S, Ca, and Mg, which suggests the absence of a direct relationship with the transport Na and Cl. In contrast, a marked decrease in the Ca concentration ([Ca]) in the cytoplasmic matrix and vacuole and an approximately two-fold increase in the P concentration ([P]) in the cytoplasmic matrix were found in S. alba, suggesting that the decrease in the [Ca] is related to the halophilic nature of S. alba (as indicated by the inward movement of Na(+) and Cl(-)). The possible roles of a Na(+)/Ca(2+) exchange mechanism in halophilism and the effect of the [P] on the metabolic activity under saline conditions are

  8. Privacy Issues of Electronic Monitoring Of Employees: A Cross-Cultural Examination Of Gender Differences

    Directory of Open Access Journals (Sweden)

    Raymond E. Taylor

    2013-07-01

    Full Text Available This manuscript presents the results of a study which examined the privacy issues of electronic monitoring of employees from a cross-cultural perspective comparing participants from Taiwan with those from the United States. The results of the study suggest that gender differences exist between Taiwanese and American respondents’ attitudes concerning privacy issues of electronic monitoring of employees. The study suggests that monitoring with notice was an important parameter in determining how privacy issues of electronic monitoring of employees were viewed by the participants.

  9. Volatiles identified from five stages of embryo development separated from a heterogeneous suspension culture of Daucus carota.

    Science.gov (United States)

    Kennedy, A H; Chamberlain, D; Wilson, G; Ryan, M F

    1991-11-01

    Five stages of embryo development were fractionated from a mature culture of Daucus carota (Gelbe Rheinsche), using a series of metal sieves. The composition of the population of embryos in each fraction was determined quantitatively from microscopic investigations. Volatiles from samples of tissue from six stages of development were trapped on activated charcoal cartridges. These volatiles, some of which may play a significant role in the interaction of the plant with the carrot root fly (Psila rosae), were analysed using gas chromatography/mass spectroscopy. The resulting chromatograms are arranged in order of embryo development. The progressive elaboration of the volatile profile reflects the increased biosynthetic capacity of the developing embryo.

  10. EDITORIAL: Colloidal suspensions Colloidal suspensions

    Science.gov (United States)

    Petukhov, Andrei; Kegel, Willem; van Duijneveldt, Jeroen

    2011-05-01

    Colloid-polymer mixtures and depletion interactions Phase stability of a reversible supramolecular polymer solution mixed with nanospheres Remco Tuinier When depletion goes critical Roberto Piazza, Stefano Buzzaccaro, Alberto Parola and Jader Colombo Tuning the demixing of colloid-polymer systems through the dispersing solvent E A G Jamie, R P A Dullens and D G A L Aarts Polydispersity effects in colloid-polymer mixtures S M Liddle, T Narayanan and W C K Poon Colloidal dynamics and crystallization Crystallization and aging in hard-sphere glasses C Valeriani, E Sanz, E Zaccarelli, W C K Poon, M E Cates and P N Pusey Real-time monitoring of complex moduli from micro-rheology Taiki Yanagishima, Daan Frenkel, Jurij Kotar and Erika Eiser Brownian motion of a self-propelled particle B ten Hagen, S van Teeffelen and H Löwen Crystallization in suspensions of hard spheres: a Monte Carlo and molecular dynamics simulation study T Schilling, S Dorosz, H J Schöpe and G Opletal Structural signature of slow dynamics and dynamic heterogeneity in two-dimensional colloidal liquids: glassy structural order Takeshi Kawasaki and Hajime Tanaka

  11. Human adipose stem cells maintain proliferative, synthetic and multipotential properties when suspension cultured as self-assembling spheroids

    International Nuclear Information System (INIS)

    Kapur, S K; Wang, X; Shang, H; Yun, S; Li, X; Feng, G; Khurgel, M; Katz, A J

    2012-01-01

    Adipose-derived stromal/stem cells (ASCs) have been gaining recognition as an extremely versatile cell source for tissue engineering. The usefulness of ASCs in biofabrication is further enhanced by our demonstration of the unique properties of these cells when they are cultured as three-dimensional cellular aggregates or spheroids. As described herein, three-dimensional formulations, or self-assembling ASC spheroids develop their own extracellular matrix that serves to increase the robustness of the cells to mechanical stresses. The composition of the extracellular matrix can be altered based on the external environment of the spheroids and these constructs can be grown in a reproducible manner and to a consistent size. The spheroid formulation helps preserve the viability and developmental plasticity of ASCs even under defined, serum-free media conditions. For the first time, we show that multiple generations of adherent ASCs produced from these spheroids retain their ability to differentiate into multiple cell/tissue types. These demonstrated properties support the idea that culture-expanded ASCs are an excellent candidate cellular material for ‘organ printing’—the approach of developing complex tissue structures from a standardized cell ‘ink’ or cell formulation. (paper)

  12. Expression of a fungal ferulic acid esterase in suspension cultures of tall fescue (Festuca arundinacea) decreases cell wall feruloylation and increases rates of cell wall digestion.

    Science.gov (United States)

    Morris, Phillip; Dalton, Sue; Langdon, Tim; Hauck, Barbara; de Buanafina, Marcia M O

    2017-01-01

    In the cell walls of grasses ferulic acid is esterified to arabinosyl residues in arabinoxylans that can then undergo oxidative coupling reactions to form ferulate dehydrodimers, trimers and oligomers which function to cross-link cell-wall polysaccharides, limiting cell wall degradability. Fungal ferulic acid esterase can release both esterified monomeric and dimeric ferulic acids from these cell wall arabinoxylans making the cell wall more susceptible to further enzymatic attack and increasing cell wall degradability. Non-embryogenic cell suspension cultures of Festuca arundinacea expressing a Aspergillus niger ferulic acid esterase ( faeA ) targeted to either the apoplast, or endoplasmic reticulum under the control of a constitutive actin promoter, or to the vacuole under the control of a soybean heat shock promoter, were established and FAE activity determined in the cells and medium during a growth cycle. Analysis of the ester-linked ferulates of the cell walls showed that all three transformed cell lines had both reduced ferulate levels and increased levels of xylanase mediated release of wall phenolics on autodigestion as well as increased rates of cell wall digestion in a simulated rumen environment, when compared to control non-transformed cells.

  13. Influence of a specific xyloglucan-nonasaccharide derived from cell walls of suspension-cultured cells of Daucus carota L. on regenerating carrot protoplasts.

    Science.gov (United States)

    Emmerling, M; Seitz, H U

    1990-09-01

    A xyloglucan oligosaccharide was isolated from cell walls of Daucus carota L. suspension-cultured cells. From analytical data (gel-permeation chromatography, thin-layer chromatography, monosaccharide analysis, methylation analysis) it can be concluded that this oligosaccharide preparation consists mainly of a nonasaccharide known as XG9 (Glc4Xyl3GalFuc). This nonasaccharide showed excellent "anti-auxin" properties in the pea-stem bioassay, with 80% inhibition of 2,4-dichlorophenoxyacetic acid (2,4-D)-induced longitudinal growth of etiolated pea stem segments at concentrations of 1-0.1 nM. Applied in nanomolar concentrations to protoplasts regenerating in a medium containing 4.52 μM 2,4-D, the nonasaccharide influenced the viability of the protoplasts and the activities of glycan synthases in vitro. The effects were similar to those achieved by the omission of 2,4-D from the regeneration medium. The composition of the regenerated cell wall was not changed significantly by the use of 2,4-D-depleted medium or the addition of XG9 to 2,4-D-containing medium.

  14. Rice suspension cultured cells are evaluated as a model system to study salt responsive networks in plants using a combined proteomic and metabolomic profiling approach.

    Science.gov (United States)

    Liu, Dawei; Ford, Kristina L; Roessner, Ute; Natera, Siria; Cassin, Andrew M; Patterson, John H; Bacic, Antony

    2013-06-01

    Salinity is one of the major abiotic stresses affecting plant productivity but surprisingly, a thorough understanding of the salt-responsive networks responsible for sustaining growth and maintaining crop yield remains a significant challenge. Rice suspension culture cells (SCCs), a single cell type, were evaluated as a model system as they provide a ready source of a homogenous cell type and avoid the complications of multicellular tissue types in planta. A combination of growth performance, and transcriptional analyses using known salt-induced genes was performed on control and 100 mM NaCl cultured cells to validate the biological system. Protein profiling was conducted using both DIGE- and iTRAQ-based proteomics approaches. In total, 106 proteins were identified in DIGE experiments and 521 proteins in iTRAQ experiments with 58 proteins common to both approaches. Metabolomic analysis provided insights into both developmental changes and salt-induced changes of rice SCCs at the metabolite level; 134 known metabolites were identified, including 30 amines and amides, 40 organic acids, 40 sugars, sugar acids and sugar alcohols, 21 fatty acids and sterols, and 3 miscellaneous compounds. Our results from proteomic and metabolomic studies indicate that the salt-responsive networks of rice SCCs are extremely complex and share some similarities with thee cellular responses observed in planta. For instance, carbohydrate and energy metabolism pathways, redox signaling pathways, auxin/indole-3-acetic acid pathways and biosynthesis pathways for osmoprotectants are all salt responsive in SCCs enabling cells to maintain cellular function under stress condition. These data are discussed in the context of our understanding of in planta salt-responses. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Video imaging of firefly luciferase activity to identify and monitor herpesvirus infection in cell culture.

    Science.gov (United States)

    Mettenleiter, T C; Gräwe, W

    1996-05-01

    Expression of reporter genes incorporated into complex viral genomes is being used increasingly to monitor virus infection in cell culture and in the organism. One of the most sensitive markers is luciferase from Photinus pyralis which catalyzes a luminescent reaction that can be traced in cell and organ extracts in luminometers. A novel method is described for monitoring Iuciferase activity after infection of cells with recombinant herpesvirus (pseudorabies virus) which carries stably and expresses luciferase using ultra high sensitive photon-counting enhanced video image acquisition. The data show that firefly luciferase activity in virus-infected cells can be monitored without destruction of the cells using video equipment for either macroscopic image acquisition or microscopy. Resolution down to a single-cell level can thereby be achieved. This method increases greatly the potential of monitoring virus infection in real time with a non-destructive highly sensitive method in cell culture and should also help to assay viral spread in the animal.

  16. Bioimpedance monitoring of 3D cell culturing-Complementary electrode configurations for enhanced spatial sensitivity

    DEFF Research Database (Denmark)

    Canali, Chiara; Heiskanen, Arto; Muhammad, Haseena Bashir

    2015-01-01

    of a bare 3D gelatin scaffold, to human mesenchymal stem cell (MSC) encapsulation and proliferation, was monitored over time. The platform consists of a large rectangular culture chamber with four embedded vertical gold plate electrodes that were exploited in two- and three terminal (2T and 3T) measurement...

  17. Production of recombinant human granulocyte macrophage-colony stimulating factor in rice cell suspension culture with a human-like N-glycan structure.

    Science.gov (United States)

    Shin, Yun-Ji; Chong, Yun-Jo; Yang, Moon-Sik; Kwon, Tae-Ho

    2011-12-01

    The rice α-amylase 3D promoter system, which is activated under sucrose-starved conditions, has emerged as a useful system for producing recombinant proteins. However, using rice as the production system for therapeutic proteins requires modifications of the N-glycosylation pattern because of the potential immunogenicity of plant-specific sugar residues. In this study, glyco-engineered rice were generated as a production host for therapeutic glycoproteins, using RNA interference (RNAi) technology to down-regulate the endogenous α-1,3-fucosyltransferase (α-1,3-FucT) and β-1,2-xylosyltransferase (β-1,2-XylT) genes. N-linked glycans from the RNAi lines were identified, and their structures were compared with those isolated from a wild-type cell suspension. The inverted-repeat chimeric RNA silencing construct of α-1,3-fucosyltransferase and β-1,2-xylosyltransferase (Δ3FT/XT)-9 glyco-engineered line with significantly reduced core α-1,3-fucosylated and/or β-1,2-xylosylated glycan structures was established. Moreover, levels of plant-specific α-1,3-fucose and/or β-1,2-xylose residues incorporated into recombinant human granulocyte/macrophage colony-stimulating factor (hGM-CSF) produced from the N44 + Δ3FT/XT-4 glyco-engineered line co-expressing ihpRNA of Δ3FT/XT and hGM-CSF were significantly decreased compared with those in the previously reported N44-08 transgenic line expressing hGM-CSF. None of the glyco-engineered lines differed from the wild type with respect to cell division, proliferation or ability to secrete proteins into the culture medium. © 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.

  18. Induction of extracellular defense-related proteins in suspension cultured-cells of Daucus carota elicited with cyclodextrins and methyl jasmonate.

    Science.gov (United States)

    Sabater-Jara, Ana B; Almagro, Lorena; Pedreño, María A

    2014-04-01

    Suspension cultured-cells (SCC) of Daucus carota were used to evaluate the effect of methyl jasmonate and cyclodextrins, separately or in combination, on the induction of defense responses, particularly the accumulation of pathogenesis-related proteins. A comparative study of the extracellular proteome (secretome) between control and elicited carrot SCC pointed to the presence of amino acid sequences homologous to glycoproteins which have inhibitory activity against the cell-wall-degrading enzymes secreted by pathogens and/or are induced when carrot cells are exposed to a pathogen elicitor. Other amino acid sequences were homologous to Leucine-Rich Repeat domain-containing proteins, which play an essential role in defense against pathogens, as well as in the recognition of microorganisms, making them important players in the innate immunity of this plant. Also, some tryptic peptides were shown to be homologous to a thaumatin-like protein, showing high specificity to abiotic stress and to different reticuline oxidase-like proteins that displayed high levels of antifungal activity, suggesting that methyl jasmonate and cyclodextrins could play a role in mediating defense-related gene product expression in SCC of D. carota. Apart from these elicitor-inducible proteins, we observed the presence of PR-proteins in both control and elicited carrot SCC, suggesting that their expression is mainly constitutive. These PR-proteins are putative class IV chitinases, which also have inhibitory activity against pathogen growth and the class III peroxidases that participate in response to environmental stress (e.g. pathogen attack and oxidative), meaning that they are involved in defense responses triggered by both biotic and abiotic factors. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  19. Vertical Deformation Monitoring of the Suspension Bridge Tower Using GNSS: A Case Study of the Forth Road Bridge in the UK

    Directory of Open Access Journals (Sweden)

    Qusen Chen

    2018-02-01

    Full Text Available The vertical deformation monitoring of a suspension bridge tower is of paramount importance to maintain the operational safety since nearly all forces are eventually transferred as the vertical stress on the tower. This paper analyses the components affecting the vertical deformation and attempts to reveal its deformation mechanism. Firstly, we designed a strategy for high-precision GNSS data processing aiming at facilitating deformation extraction and analysis. Then, 33 months of vertical deformation time series of the southern tower of the Forth Road Bridge (FRB in the UK were processed, and the accurate subsidence and the parameters of seasonal signals were estimated based on a classic function model that has been widely studied to analyse GNSS coordinate time series. We found that the subsidence rate is about 4.7 mm/year, with 0.1 mm uncertainty. Meanwhile, a 15-month meteorological dataset was utilised with a thermal expansion model (TEM to explain the effects of seasonal signals on tower deformation. The amplitude of the annual signals correlated quite well that obtained by the TEM, with the consistency reaching 98.9%, demonstrating that the thermal effect contributes significantly to the annual signals. The amplitude of daily signals displays poor consistency with the ambient temperature data. However, the phase variation tendencies between the daily signals of the vertical deformation and the ambient temperature are highly consistent after February 2016. Finally, the potential contribution of the North Atlantic Drift (NAD to the characteristics of annual and daily signals is discussed because of the special geographical location of the FRB. Meanwhile, this paper emphasizes the importance of collecting more detailed meteorological and other loading data for the investigation of the vertical deformation mechanism of the bridge towers over time with the support of GNSS.

  20. Stress Response Monitoring of Photoautotrophic Higher Plant Suspension Cultures by Fluorescence Imaging for High-Throughput Toxic Compound Screening

    Czech Academy of Sciences Publication Activity Database

    Segečová, Anna; Červený, Jan; Roitsch, Thomas

    2017-01-01

    Roč. 8, č. 6 (2017), s. 678-692 ISSN 2152-2197 R&D Projects: GA MŠk(CZ) LO1415; GA ČR(CZ) GA15-17367S; GA MŠk(CZ) LM2015055 Institutional support: RVO:67179843 Keywords : Toxins * Toxicants * Ecotoxicology * PAM Chlorophyll Fluorescence Subject RIV: CE - Biochemistry OBOR OECD: Biochemistry and molecular biology

  1. Idaho National Laboratory Cultural Resource Monitoring Report for Fiscal Year 2016

    Energy Technology Data Exchange (ETDEWEB)

    Gilbert, Hollie Kae [Idaho National Lab. (INL), Idaho Falls, ID (United States); Holmer, Marie Pilkington [Idaho National Lab. (INL), Idaho Falls, ID (United States); Olson, Christina Liegh [Idaho National Lab. (INL), Idaho Falls, ID (United States); Pace, Brenda Ringe [Idaho National Lab. (INL), Idaho Falls, ID (United States)

    2016-11-01

    This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during fiscal year (FY) 2016. Overall monitoring included surveillance of the following 23 individual cultural resource localities: two locations with human remains, one of which is also a cave; seven additional caves; six prehistoric archaeological sites; four historic archaeological sites; one historic trail; Experimental Breeder Reactor I (EBR-I), a National Historic Landmark; Aircraft Nuclear Propulsion (ANP) objects located at EBR-I; and one Arco Naval Proving Ground (NPG) property, CF-633 and related objects and structures. Several INL work processes and projects were also monitored to confirm compliance with original INL CRM recommendations and assess the effects of ongoing work. On one occasion, ground disturbing activities within the boundaries of the Critical Infrastructure Test Range Complex (CITRC) were observed by INL CRM staff prepared to respond to any additional finds of Native American human remains. Additionally, the CRM office was notified during two Trespass Investigations conducted by INL Security. Most of the cultural resources monitored in FY 2016 exhibited no adverse impacts, resulting in Type 1 impact assessments. However, Type 2 impacts were noted five times. Three previously reported Type 2 impacts were once again documented at the EBR-I National Historic Landmark, including spalling and deterioration of bricks due to inadequate drainage, minimal maintenance, and rodent infestation. The ANP engines and locomotive on display at the EBR-I Visitors Center also exhibited impacts related to long term exposure. Finally, most of the Arco NPG properties monitored at Central Facilities Area exhibited problems with lack of timely and appropriate maintenance as well as inadequate drainage. No new Type 3 or Type 4 impacts that adversely affected significant cultural resources and threatened National

  2. Pericellular oxygen monitoring with integrated sensor chips for reproducible cell culture experiments.

    Science.gov (United States)

    Kieninger, J; Aravindalochanan, K; Sandvik, J A; Pettersen, E O; Urban, G A

    2014-04-01

    Here we present an application, in two tumour cell lines, based on the Sensing Cell Culture Flask system as a cell culture monitoring tool for pericellular oxygen sensing. T-47D (human breast cancer) and T98G (human brain cancer) cells were cultured either in atmospheric air or in a glove-box set at 4% oxygen, in both cases with 5% CO2 in the gas phase. Pericellular oxygen tension was measured with the help of an integrated sensor chip comprising oxygen sensor arrays. Obtained results illustrate variation of pericellular oxygen tension in attached cells covered by stagnant medium. Independent of incubation conditions, low pericellular oxygen concentration levels, usually associated with hypoxia, were found in dense cell cultures. Respiration alone brought pericellular oxygen concentration down to levels which could activate hypoxia-sensing regulatory processes in cultures believed to be aerobic. Cells in culture believed to experience conditions of mild hypoxia may, in reality, experience severe hypoxia. This would lead to incorrect assumptions and suggests that pericellular oxygen concentration readings are of great importance to obtain reproducible results when dealing with hypoxic and normoxic (aerobic) incubation conditions. The Sensing Cell Culture Flask system allows continuous monitoring of pericellular oxygen concentration with outstanding long-term stability and no need for recalibration during cell culture experiments. The sensor is integrated into the flask bottom, thus in direct contact with attached cells. No additional equipment needs to be inserted into the flask during culturing. Transparency of the electrochemical sensor chip allows optical inspection of cells attached on top of the sensor. © 2014 John Wiley & Sons Ltd.

  3. Cell division in Escherichia coli cultures monitored at single cell resolution

    Directory of Open Access Journals (Sweden)

    Luidalepp Hannes

    2008-04-01

    Full Text Available Abstract Background A fundamental characteristic of cells is the ability to divide. To date, most parameters of bacterial cultures, including cell division, have been measured as cell population averages, assuming that all bacteria divide at a uniform rate. Results We monitored the division of individual cells in Escherichia coli cultures during different growth phases. Our experiments are based on the dilution of green fluorescent protein (GFP upon cell division, monitored by flow cytometry. The results show that the vast majority of E. coli cells in exponentially growing cultures divided uniformly. In cultures that had been in stationary phase up to four days, no cell division was observed. However, upon dilution of stationary phase culture into fresh medium, two subpopulations of cells emerged: one that started dividing and another that did not. These populations were detectable by GFP dilution and displayed different side scatter parameters in flow cytometry. Further analysis showed that bacteria in the non-growing subpopulation were not dead, neither was the difference in growth capacity reducible to differences in stationary phase-specific gene expression since we observed uniform expression of several stress-related promoters. The presence of non-growing persisters, temporarily dormant bacteria that are tolerant to antibiotics, has previously been described within growing bacterial populations. Using the GFP dilution method combined with cell sorting, we showed that ampicillin lyses growing bacteria while non-growing bacteria retain viability and that some of them restart growth after the ampicillin is removed. Thus, our method enables persisters to be monitored even in liquid cultures of wild type strains in which persister formation has low frequency. Conclusion In principle, the approaches developed here could be used to detect differences in cell division in response to different environmental conditions and in cultures of unicellular

  4. A multi-disciplinary approach for the structural monitoring of Cultural Heritages in a seismic area

    Science.gov (United States)

    Fabrizia Buongiorno, Maria; Musacchio, Massimo; Guerra, Ignazio; Porco, Giacinto; Stramondo, Salvatore; Casula, Giuseppe; Caserta, Arrigo; Speranza, Fabio; Doumaz, Fawzi; Giovanna Bianchi, Maria; Luzi, Guido; Ilaria Pannaccione Apa, Maria; Montuori, Antonio; Gaudiosi, Iolanda; Vecchio, Antonio; Gervasi, Anna; Bonali, Elena; Romano, Dolores; Falcone, Sergio; La Piana, Carmelo

    2014-05-01

    In the recent years, the concepts of seismic risk vulnerability and structural health monitoring have become very important topics in the field of both structural and civil engineering for the identification of appropriate risk indicators and risk assessment methodologies in Cultural Heritages monitoring. The latter, which includes objects, building and sites with historical, architectural and/or engineering relevance, concerns the management, the preservation and the maintenance of the heritages within their surrounding environmental context, in response to climate changes and natural hazards (e.g. seismic, volcanic, landslides and flooding hazards). Within such a framework, the complexity and the great number of variables to be considered require a multi-disciplinary approach including strategies, methodologies and tools able to provide an effective monitoring of Cultural Heritages form both scientific and operational viewpoints. Based on this rationale, in this study, an advanced, technological and operationally-oriented approach is presented and tested, which enables measuring and monitoring Cultural Heritage conservation state and geophysical/geological setting of the area, in order to mitigate the seismic risk of the historical public goods at different spatial scales*. The integration between classical geophysical methods with new emerging sensing techniques enables a multi-depth, multi-resolution, and multi-scale monitoring in both space and time. An integrated system of methodologies, instrumentation and data-processing approaches for non-destructive Cultural Heritage investigations is proposed, which concerns, in detail, the analysis of seismogenetic sources, the geological-geotechnical setting of the area and site seismic effects evaluation, proximal remote sensing techniques (e.g. terrestrial laser scanner, ground-based radar systems, thermal cameras), high-resolution aerial and satellite-based remote sensing methodologies (e.g. aeromagnetic surveys

  5. Monitoring Campylobacter in the poultry production chain - From culture to genes and beyond

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hasseldam; Bhunia, Arun; Engvall, Eva Olsson

    2015-01-01

    numbers of samples. The ultimate goal for rapid monitoring of Campylobacter is to prevent contaminated meat from entering the food market. Currently, real-time PCR is fulfilling abovementioned criteria to a certain extent. Further development of real-time PCR, microarray PCR, miniaturized biosensors......Improved monitoring tools are important for the control of Campylobacter bacteria in poultry production. Standardized reference culture methods issued by national and international standardization organizations are time-consuming, cumbersome and not amenable to automation for screening of large......, chromatographic techniques and DNA sequencing can improve our monitoring capacity at a lower cost. Combined with innovative sampling and sample treatment, these techniques could become realistic options for on-farm and liquid-sample monitoring at slaughterhouses....

  6. Monitoring Campylobacter in the poultry production chain—from culture to genes and beyond.

    Science.gov (United States)

    Josefsen, Mathilde H; Bhunia, Arun K; Engvall, Eva Olsson; Fachmann, Mette S R; Hoorfar, Jeffrey

    2015-05-01

    Improved monitoring tools are important for the control of Campylobacter bacteria in poultry production. Standardized reference culture methods issued by national and international standardization organizations are time-consuming, cumbersome and not amenable to automation for screening of large numbers of samples. The ultimate goal for rapid monitoring of Campylobacter is to prevent contaminated meat from entering the food market. Currently, real-time PCR is fulfilling abovementioned criteria to a certain extent. Further development of real-time PCR, microarray PCR, miniaturized biosensors, chromatographic techniques and DNA sequencing can improve our monitoring capacity at a lower cost. Combined with innovative sampling and sample treatment, these techniques could become realistic options for on-farm and liquid-sample monitoring at slaughterhouses. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. [Quality indicators for blood culture: three years of monitoring at a university hospital in Chile].

    Science.gov (United States)

    Guzmán, Ana María; Sánchez, Tomás; de la Barra, Ricardo

    2012-08-01

    Blood culture is considered the "gold standard" for the diagnosis of bacteremia, critical condition with high morbidity and mortality. Because of its importance, it is estimated that the blood culture is a critical test that requires close monitoring on the quality with which the process is performed. The objective of this work is to show the results of the monitoring carried out during the past three years, of 5 quality indicators of blood cultures in the laboratory of the Hospital Clínico de la Pontificia Universidad Católica de Chile, considering pre-analytical, analytical and post-analytical aspects. In the 3 years monitored the mean contamination was 0,7%, 46% of adult bottles had adequate volume, match between Gram stain with final identification was 99.4%, 100% of correct participations were achieved in surveys of external quality control and Gram staining notification before 1 hour was 88.7%. With regard to proposed aims, in 2011 the laboratory complies with all, except the percentage of bottles with appropriate volume of blood inoculated. This indicator is very low and should be corrected as soon as possible since it is known that it is an important condition for optimum performance of blood cultures.

  8. Multifunctional sensing membrane-based platform for tissue or cell culturing and monitoring

    DEFF Research Database (Denmark)

    2014-01-01

    The present application discloses a water-permeable sensor membrane comprising i) a first layer of a conductive material defining at least one electrode and having a thickness of 0.1-,000 [mu]m; ii) a second layer of a nanostructure material build on the first layer; and iii) a third, topmost......, layer of a conducting polymer material defining at least one electrode and having a thickness of 0.001-1.0 [mu]m. The application also discloses a tissue or cell culture sample monitoring assembly comprising a sensor assembly and a tissue sample or a cell culture sample arranged on top of the third...

  9. Toward an integrated system concept for monitoring and evaluation of safety culture

    International Nuclear Information System (INIS)

    Makino, Maomi; Sakaue, Takeharu

    2004-01-01

    The concept of ''nuclear safety culture'' has been advocated and has been much discussed internationally by INSAG (The International Nuclear Safety Advisory Group) under IAEA (the International Atomic Energy Agency) and other institutions since Chernobyl accident. On the safety front, Japan had maintained an excellent track record in nuclear power operations throughout the 1990s. However, there have been a series of new type of problems strongly implying degradation of safety culture, e.g., Monju accident, fire and explosion accident at an Asphalt Solidification Process Facility at Tokai, falsification of annealing data at nuclear power plants (NPP), another data falsification for transport cask of spent fuel and JCO criticality accident. Then the TEPCO (Tokyo Electric Power Company) issue was revealed in 2002. Triggered by this issue, the Nuclear and Industrial Safety Agency (NISA) has been implementing a variety of improvements, one of which was the establishment of a study group in 2003, which invited experts from other fields as well as from nuclear-related industries, to study on how to implement safety culture sufficiently and possible recommendations. Subjects such as the followings piled in the study report will indicate leading keys in case it is going to realize such efforts: ''Foundation of safety culture is a quality management'' and ''Realistic and scientific technique is necessary for the evaluation of safety culture''. In order to respond to these requests, JNES have been advancing the development toward an Integrated System Concept for Monitoring and Evaluation of Safety Culture. This paper describes the outline of the study results reported by the study group and then introduces one of subsystems, SCEST, structuring the integrated system concept for Monitoring and Evaluation of Safety Culture. (author)

  10. Conducting Polymer Scaffolds for Hosting and Monitoring 3D Cell Culture

    KAUST Repository

    Inal, Sahika

    2017-05-03

    This work reports the design of a live-cell monitoring platform based on a macroporous scaffold of a conducting polymer, poly(3,4-ethylene dioxythiophene):poly(styrenesulfonate). The conducting polymer scaffolds support 3D cell cultures due to their biocompatibility and tissue-like elasticity, which can be manipulated by inclusion of biopolymers such as collagen. Integration of a media perfusion tube inside the scaffold enables homogenous cell spreading and fluid transport throughout the scaffold, ensuring long term cell viability. This also allows for co-culture of multiple cell types inside the scaffold. The inclusion of cells within the porous architecture affects the impedance of the electrically conducting polymer network and, thus, is utilized as an in situ tool to monitor cell growth. Therefore, while being an integral part of the 3D tissue, the conducting polymer is an active component, enhancing the tissue function, and forming the basis for a bioelectronic device with integrated sensing capability.

  11. The efficient method for simultaneous monitoring of the culturable as well as nonculturable airborne microorganisms.

    Directory of Open Access Journals (Sweden)

    Barbara Hubad

    Full Text Available Cultivation-based microbiological methods are a gold standard for monitoring of airborne micro-organisms to determine the occupational exposure levels or transmission paths of a particular infectious agent. Some highly contagious microorganisms are not easily culturable but it is becoming evident that cultivation and molecular methods are complementary and in these cases highly relevant. We report a simple and efficient method for sampling and analyzing airborne bacteria with an impactor-type high-flow-rate portable air sampler, currently used for monitoring culturable bacteria or fungi. A method is reported for extraction of nucleic acids from impacted cells without prior cultivation and using agarose as a sampling matrix. The DNA extraction efficiency was determined in spiked samples and, samples taken from a wastewater treatment plant and an alpine area. The abundance, diversity and quantity of total bacteria were analysed by a quantitative polymerase chain reaction, and by construction and analysis of clone libraries. The method does not interfere with downstream PCR analysis and can cover the gap between traditional culture and molecular techniques of bioaerosol monitoring.

  12. The Contribution of Earth Observation Technologies to Monitoring Strategies of Cultural Landscapes and Sites

    Science.gov (United States)

    Cuca, B.

    2017-08-01

    Coupling of Climate change effects with management and protection of cultural and natural heritage has been brought to the attention of policy makers since several years. On the worldwide level, UNESCO has identified several phenomena as the major geo-hazards possibly induced by climate change and their possible hazardous impact to natural and cultural heritage: Hurricane, storms; Sea-level rise; Erosion; Flooding; Rainfall increase; Drought; Desertification and Rise in temperature. The same document further referrers to satellite Remote Sensing (EO) as one of the valuable tools, useful for development of "professional monitoring strategies". More recently, other studies have highlighted on the impact of climate change effects on tourism, an economic sector related to build environment and traditionally linked to heritage. The results suggest that, in case of emergency the concrete threat could be given by the hazardous event itself; in case of ordinary administration, however, the threat seems to be a "hazardous attitude" towards cultural assets that could lead to inadequate maintenance and thus to a risk of an improper management of cultural heritage sites. This paper aims to illustrate potential benefits that advancements of Earth Observation technologies can bring to the domain of monitoring landscape heritage and to the management strategies, including practices of preventive maintenance. The attempt here is to raise awareness on the importance of integrating satellite remote sensing imagery and the deriving products with other geospatial information (even geo-referenced historic maps) for a more complete insight on the environmental dynamics of landscapes.

  13. Application of near-infrared spectroscopy for monitoring and control of cell culture and fermentation

    DEFF Research Database (Denmark)

    Cervera Padrell, Albert Emili; Petersen, Nanna; Eliasson Lantz, Anna

    2009-01-01

    Near-infrared (NIR) spectroscopy can potentially provide on-line information on substrate, biomass, product, and metabolite concentrations in fermentation processes, which could be useful for improved monitoring or control. However, several factors can negatively influence the quality...... of chemometric models built for interpretation of the spectra, thus impairing the analyte concentration predictions. The aim of this review was to provide an overview of necessary conditions and challenges that one has to face when developing a NIR application for monitoring of cell culture or fermentation...... fusion), and the description of process trajectories. On the basis of the review, we conclude that acceptance of NIR spectroscopy as a standard monitoring tool by the fermentation industry will necessitate considerably more on-line studies using industrially relevant—and highly challenging...

  14. Whole-cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production.

    Science.gov (United States)

    Goers, Lisa; Ainsworth, Catherine; Goey, Cher Hui; Kontoravdi, Cleo; Freemont, Paul S; Polizzi, Karen M

    2017-06-01

    Many high-value added recombinant proteins, such as therapeutic glycoproteins, are produced using mammalian cell cultures. In order to optimize the productivity of these cultures it is important to monitor cellular metabolism, for example the utilization of nutrients and the accumulation of metabolic waste products. One metabolic waste product of interest is lactic acid (lactate), overaccumulation of which can decrease cellular growth and protein production. Current methods for the detection of lactate are limited in terms of cost, sensitivity, and robustness. Therefore, we developed a whole-cell Escherichia coli lactate biosensor based on the lldPRD operon and successfully used it to monitor lactate concentration in mammalian cell cultures. Using real samples and analytical validation we demonstrate that our biosensor can be used for absolute quantification of metabolites in complex samples with high accuracy, sensitivity, and robustness. Importantly, our whole-cell biosensor was able to detect lactate at concentrations more than two orders of magnitude lower than the industry standard method, making it useful for monitoring lactate concentrations in early phase culture. Given the importance of lactate in a variety of both industrial and clinical contexts we anticipate that our whole-cell biosensor can be used to address a range of interesting biological questions. It also serves as a blueprint for how to capitalize on the wealth of genetic operons for metabolite sensing available in nature for the development of other whole-cell biosensors. Biotechnol. Bioeng. 2017;114: 1290-1300. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

  15. Dual Monitoring of Secretion and ATP Levels during Chondrogenesis Using Perfusion Culture-Combined Bioluminescence Monitoring System

    Directory of Open Access Journals (Sweden)

    Hyuck Joon Kwon

    2015-01-01

    Full Text Available Skeletal pattern formation in limb development depends on prechondrogenic condensation which prefigures the cartilage template. However, although morphogens such as TGF-βs and BMPs have been known to play essential roles in skeletal patterning, how the morphogens induce prechondrogenic cells to aggregate and determine patterns of cartilage elements has remained unclear. Our previous study reported that ATP oscillations are induced during chondrogenesis. This result suggests the possibility that ATP oscillations lead to the oscillatory secretion of morphogens, due to the fact that secretion process requires ATP. To examine the correlation between ATP oscillations and secretion levels of morphogens, we have developed perfusion culture-combined bioluminescence monitoring system to simultaneously monitor intracellular ATP levels and secretion levels. Using this system, we found that secretory activity oscillates in phase with ATP oscillations and that secretion levels of TGF-β1 and BMP2 oscillate during chondrogenesis. The oscillatory secretion of the morphogens would contribute to amplifying the fluctuation of the morphogens, underlie the spatial patterning of morphogens, and consequently lead to skeletal pattern formation.

  16. Monitoring cell culture media degradation using surface enhanced Raman scattering (SERS) spectroscopy.

    Science.gov (United States)

    Calvet, Amandine; Ryder, Alan G

    2014-08-20

    The quality of the cell culture media used in biopharmaceutical manufacturing is a crucial factor affecting bioprocess performance and the quality of the final product. Due to their complex composition these media are inherently unstable, and significant compositional variations can occur particularly when in the prepared liquid state. For example photo-degradation of cell culture media can have adverse effects on cell viability and thus process performance. There is therefore, from quality control, quality assurance and process management view points, an urgent demand for the development of rapid and inexpensive tools for the stability monitoring of these complex mixtures. Spectroscopic methods, based on fluorescence or Raman measurements, have now become viable alternatives to more time-consuming and expensive (on a unit analysis cost) chromatographic and/or mass spectrometry based methods for routine analysis of media. Here we demonstrate the application of surface enhanced Raman scattering (SERS) spectroscopy for the simple, fast, analysis of cell culture media degradation. Once stringent reproducibility controls are implemented, chemometric data analysis methods can then be used to rapidly monitor the compositional changes in chemically defined media. SERS shows clearly that even when media are stored at low temperature (2-8°C) and in the dark, significant chemical changes occur, particularly with regard to cysteine/cystine concentration. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Genome profiling (GP) as an effective tool for monitoring culture collections: a case study with Trichosporon.

    Science.gov (United States)

    Hamano, Keiichi; Ueno-Tsuji, Sachika; Tanaka, Reiko; Suzuki, Motofumi; Nishimura, Kazuko; Nishigaki, Koichi

    2012-05-01

    Species identification and classification of a large number of microbes are essential and heavy workloads in culture collections and relevant laboratories. The identification of species usually requires different methods depending on species. Therefore, the development of a method which is simple and applicable to any organisms will lessen the burdens, increase the reliability of databases and thus enhance the science on microbes. The genome profiling (GP) method, developed previously, was found effective in monitoring authenticities of all strains/species tested in culture collections and expectedly various species, which was shown by applying the GP and the conventional sequencing methods to identifying and classifying species/strains belonging to the genus Trichosporon (38 strains; 16 species). Small differences between strains (11 strains of Trichosporon asahii and 4 strains of Trichosporon coremiiforme) can be reliably discriminated by GP, which was unsuccessful in the conventional sequencing approach. Importantly, seven possible false-assignments contained in the database were all pointed out by the GP method with near-perfect correctness, showing the power of the GP method.GP was shown to be a potent tool for rapidly and correctly monitoring species and strains of fungi in culture collections owing to its simple and informative natures. Copyright © 2012 Elsevier B.V. All rights reserved.

  18. Idaho National Laboratory Cultural Resource Monitoring Report for Fiscal Year 2007

    Energy Technology Data Exchange (ETDEWEB)

    Brenda R. Pace

    2007-10-01

    This report describes the cultural resource monitoring activities of the Idaho National Laboratory’s (INL) Cultural Resource Management (CRM) Office during fiscal year 2007 (FY 2007). In FY 2007, 40 localities were revisited: two locations of heightened Shoshone-Bannock tribal sensitivity, four caves, three butte/craters, twelve prehistoric archaeological sites, two historic stage stations, nine historic homesteads, a portion of Goodale’s Cutoff of the Oregon Trail, a portion of historic trail T-16, one World War II dump, four buildings from the World War II period, and Experimental Breeder Reactor –I, a modern scientific facility and National Historic Landmark. Several INL project areas were also monitored in FY 2007. This included direct observation of ground disturbing activities within the Power Burst Facility (PBF, now designated as the Critical Infrastructure Test Range Complex – CITRC), backfilling operations associated with backhoe trenches along the Big Lost River, and geophysical surveys designed to pinpoint subsurface unexploded ordnance in the vicinity of the Naval Ordnance Disposal Area. Surprise checks were also made to three ongoing INL projects to ensure compliance with INL CRM Office recommendations to avoid impacts to cultural resources. Although some impacts were documented, no significant adverse effects that would threaten the National Register eligibility of any resource were observed at any location.

  19. An electrochemical approach to monitor pH change in agar media during plant tissue culture.

    Science.gov (United States)

    Wang, Min; Ha, Yang

    2007-05-15

    In this work, metal oxide microelectrodes were developed to monitor pH change in agar media during plant tissue culture. An antimony wire was produced by a new approach "capillary melt method". The surface of the obtained antimony wire was oxidized in a potassium nitrate melt to fabricate an antimony oxide film for pH sensing. Characterization results show that the oxide layer grown on the wire surface consists of Sb(2)O(3) crystal phase. The sensing response, open-circuit potential, of the electrode has a good linear relationship (R(2)=1.00) with pH value of the test solution. Adding organic compounds into the test media would not affect the linear relationship, although the slope of the lines varied with different ingredients added. The antimony oxide electrodes were employed to continuously monitor pH change of agar culture media during a 2-week plant tissue culture of Dendrobium candidum. The antimony oxide electrode fabricated this way has the advantages of low cost, easy fabrication, fast response, and almost no contamination introduced into the system. It would be suitable for in situ and continuous pH measurement in many bio applications.

  20. EOP Acoustic tagging and monitorings of cultured and wild juvenile crimson jobfish (Pristipomoides filamentosus) in a nursery habitat

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Raw data from Vemco receivers that monitored the Kaneohe, Oahu nursery grounds while tagged juvenile snapper were released in 2006 (cultured) and 2007 (wild). Also...

  1. 2,3,5,4′- Tetrahydroxystilbene-2-O-β-D-glycoside Biosynthesis by Suspension Cells Cultures of Polygonum multiflorum Thunb and Production Enhancement by Methyl Jasmonate and Salicylic Acid

    Directory of Open Access Journals (Sweden)

    Li Shao

    2012-02-01

    Full Text Available Friable calli of Polygonum multiflorum Thunb have been induced in MS medium supplemented with 6-benzylaminopurine (6-BA and kinetin (KT. Suspension cultures were initiated from friable calli by inoculating calli in liquid MS medium in shake flasks in the dark and 25 °C on an orbital shaker at 100 rpm. The maximum dry weight (DW, 7.85 g/L and 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glycoside (THSG, 56.39 mg/L of suspension cells was obtained in MS medium after 16 days culture. Both methyl jasmonate (MeJA and salicylic acid (SA could increase THSG production. The most appropriate concentration of MeJA was 100 μmol/L in MS medium, in which concentration THSG content reached the maximum value of 147.79 mg/L, which represented a 162.36% increase compared to that of the control (56.33 mg/L. The most appropriate concentration of SA was 125 μmol/L in MS medium, at which concentration THSG content reached its maximum value of 116.43 mg/L, a 106.69% increase compared to that of the control (56.33 mg/L.

  2. 2,3,5,4'- tetrahydroxystilbene-2-O-β-D-glycoside biosynthesis by suspension cells cultures of Polygonum multiflorum Thunb and production enhancement by methyl jasmonate and salicylic acid.

    Science.gov (United States)

    Shao, Li; Zhao, Shu-Jin; Cui, Tang-Bing; Liu, Zhong-Yu; Zhao, Wei

    2012-02-22

    Friable calli of Polygonum multiflorum Thunb have been induced in MS medium supplemented with 6-benzylaminopurine (6-BA) and kinetin (KT). Suspension cultures were initiated from friable calli by inoculating calli in liquid MS medium in shake flasks in the dark and 25 °C on an orbital shaker at 100 rpm. The maximum dry weight (DW, 7.85 g/L) and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glycoside (THSG, 56.39 mg/L) of suspension cells was obtained in MS medium after 16 days culture. Both methyl jasmonate (MeJA) and salicylic acid (SA) could increase THSG production. The most appropriate concentration of MeJA was 100 μmol/L in MS medium, in which concentration THSG content reached the maximum value of 147.79 mg/L, which represented a 162.36% increase compared to that of the control (56.33 mg/L). The most appropriate concentration of SA was 125 μmol/L in MS medium, at which concentration THSG content reached its maximum value of 116.43 mg/L, a 106.69% increase compared to that of the control (56.33 mg/L).

  3. Particle size distribution and total solids suspension in samples monitoring of capturing water for optimization of water injection filtration system; Monitoramento da quantidade de particulas e do total de solidos em suspensao em amostras de agua de captacao

    Energy Technology Data Exchange (ETDEWEB)

    Ramalhao, Adriano Gorga; Seno, Carlos Eduardo; Ribeiro, Alice [3M do Brasil, Sumare, SP (Brazil)

    2008-07-01

    There is a wide variation in the amount of particulate material in sea water by a great number of reasons. The most well-known contaminant is the organic material derived from seaweed or fish spawning causing seasonally sensitive variations in the water quality treated and injected for enhance oil recovery. This paper presents the results of one year the water monitoring form water sampled at 30 meters deep in the Roncador field, which is located 125 km from the coast with a depth of 1290 meters. It was observed the water seasonal variation with peaks in summer and winter. The monitoring was done through particle counting and distribution analysis and total solids in suspension. It was noted that even in peak with largest amount of particles and greater quantity of solid in suspension the particles had remained concentrated in the range bellow 25 {mu}m. For that reason the life of final filter elements may vary and pre-filters are many times ineffective and sometimes even bypassed due to frequent clogging and not to do the protecting job of the final filter. (author)

  4. Suspension Trauma / Orthostatic Intolerance

    Science.gov (United States)

    ... Technology Assessment Printer Friendly Version Suspension Trauma/Orthostatic Intolerance Safety and Health Information Bulletin SHIB 03-24- ... with important information about the hazards of orthostatic intolerance and suspension trauma when using fall arrest systems. ...

  5. Monitoring and robust adaptive control of fed-batch cultures of microorganisms exhibiting overflow metabolism [abstract

    Directory of Open Access Journals (Sweden)

    Vande Wouwer, A.

    2010-01-01

    Full Text Available Overflow metabolism characterizes cells strains that are likely to produce inhibiting by-products resulting from an excess of substrate feeding and a saturated respiratory capacity. The critical substrate level separating the two different metabolic pathways is generally not well defined. Monitoring of this kind of cultures, going from model identification to state estimation, is first discussed. Then, a review of control techniques which all aim at maximizing the cell productivity of fed-batch fermentations is presented. Two main adaptive control strategies, one using an estimation of the critical substrate level as set-point and another regulating the by-product concentration, are proposed. Finally, experimental investigations of an adaptive RST control scheme using the observer polynomial for the regulation of the ethanol concentration in Saccharomyces cerevisiae fed-batch cultures ranging from laboratory to industrial scales, are also presented.

  6. Multichannel Bipotentiostat Integrated With a Microfluidic Platform for Electrochemical Real-Time Monitoring of Cell Cultures

    DEFF Research Database (Denmark)

    Vergani, Marco; Carminati, Marco; Ferrari, Giorgio

    2012-01-01

    An electrochemical detection system specifically designed for multi-parameter real-time monitoring of stem cell culturing/differentiation in a microfluidic system is presented. It is composed of a very compact 24-channel electronic board, compatible with arrays of microelectrodes and coupled...... to a microfluidic cell culture system. A versatile data acquisition software enables performing amperometry, cyclic voltammetry and impedance spectroscopy in each of the 12 independent chambers over a 100 kHz bandwidth with current resolution down to 5 pA for 100 ms measuring time. The design of the platform, its...... realization and experimental characterization are reported, with emphasis on the analysis of impact of input capacitance (i.e., microelectrode size) and microfluidic pump operation on current noise. Programmable sequences of successive injections of analytes (ferricyanide and dopamine) and rinsing buffer...

  7. Regulated bioluminescence as a tool for bioremediation process monitoring and control of bacterial cultures

    Science.gov (United States)

    Burlage, Robert S.; Heitzer, Armin; Digrazia, Philip M.

    1991-01-01

    An effective on-line monitoring technique for toxic waste bioremediation using bioluminescent microorganisms has shown great potential for the description and optimization of biological processes. The lux genes of the bacterium Vibrio fischeri are used by this species to produce visible light. The lux genes can be genetically fused to the control region of a catabolic gene, with the result that bioluminescence is produced whenever the catabolic gene is induced. Thus the detection of light from a sample indicates that genetic expression from a specific gene is occurring. This technique was used to monitor biodegradation of specific contaminants from waste sites. For these studies, fusions between the lux genes and the operons for naphthalene and toluene/xylene degradation were constructed. Strains carrying one of these fusions respond sensitively and specifically to target substrates. Bioluminescence from these cultures can be rapidly measured in a nondestructive and noninvasive manner. The potential for this technique in this and other biological systems is discussed.

  8. Using white noise to gate organic transistors for dynamic monitoring of cultured cell layers.

    Science.gov (United States)

    Rivnay, Jonathan; Leleux, Pierre; Hama, Adel; Ramuz, Marc; Huerta, Miriam; Malliaras, George G; Owens, Roisin M

    2015-06-26

    Impedance sensing of biological systems allows for monitoring of cell and tissue properties, including cell-substrate attachment, layer confluence, and the "tightness" of an epithelial tissue. These properties are critical for electrical detection of tissue health and viability in applications such as toxicological screening. Organic transistors based on conducting polymers offer a promising route to efficiently transduce ionic currents to attain high quality impedance spectra, but collection of complete impedance spectra can be time consuming (minutes). By applying uniform white noise at the gate of an organic electrochemical transistor (OECT), and measuring the resulting current noise, we are able to dynamically monitor the impedance and thus integrity of cultured epithelial monolayers. We show that noise sourcing can be used to track rapid monolayer disruption due to compounds which interfere with dynamic polymerization events crucial for maintaining cytoskeletal integrity, and to resolve sub-second alterations to the monolayer integrity.

  9. Selection of microbial populations degrading recalcitrant hydrocarbons of gasoline by monitoring of culture-headspace composition.

    Science.gov (United States)

    Solano-Serena, F; Marchal, R; Lebeault, J M; Vandecasteele, J P

    2000-01-01

    A methodology was devised and was found useful for the selection of populations degrading recalcitrant hydrocarbons. The work was part of a programme aiming at developing knowledge of the intrinsic capacities of autochtonous microflorae of the environment for gasoline biodegradation. The methodology involved monitoring the progress of degradation in enrichment liquid cultures on the selected hydrocarbon by gas chromatographic analysis of CO2 production and O2 consumption. Populations degrading in particular o-xylene, 1,2,4-trimethylbenzene, cyclohexane were obtained. Concerning 2,2,4-trimethylpentane (isooctane), one microflora (and a pure strain derived from it) growing on this hydrocarbon were obtained from gasoline-polluted water.

  10. THE CONTRIBUTION OF EARTH OBSERVATION TECHNOLOGIES TO MONITORING STRATEGIES OF CULTURAL LANDSCAPES AND SITES

    Directory of Open Access Journals (Sweden)

    B. Cuca

    2017-08-01

    Full Text Available Coupling of Climate change effects with management and protection of cultural and natural heritage has been brought to the attention of policy makers since several years. On the worldwide level, UNESCO has identified several phenomena as the major geo-hazards possibly induced by climate change and their possible hazardous impact to natural and cultural heritage: Hurricane, storms; Sea-level rise; Erosion; Flooding; Rainfall increase; Drought; Desertification and Rise in temperature. The same document further referrers to satellite Remote Sensing (EO as one of the valuable tools, useful for development of “professional monitoring strategies”. More recently, other studies have highlighted on the impact of climate change effects on tourism, an economic sector related to build environment and traditionally linked to heritage. The results suggest that, in case of emergency the concrete threat could be given by the hazardous event itself; in case of ordinary administration, however, the threat seems to be a “hazardous attitude” towards cultural assets that could lead to inadequate maintenance and thus to a risk of an improper management of cultural heritage sites. This paper aims to illustrate potential benefits that advancements of Earth Observation technologies can bring to the domain of monitoring landscape heritage and to the management strategies, including practices of preventive maintenance. The attempt here is to raise awareness on the importance of integrating satellite remote sensing imagery and the deriving products with other geospatial information (even geo-referenced historic maps for a more complete insight on the environmental dynamics of landscapes.

  11. Relationship between IQ, cultural intelligence and self-monitoring in the students of Birjand University of Medical Sciences

    Directory of Open Access Journals (Sweden)

    Aliakbar Esmaeili

    2016-09-01

    Full Text Available Background and Aim: Intelligence quotient (IQ, cultural intelligence, and self-monitoring are among important and influential parameters in learning-teaching process of students. Thus, the current study examined the relationship between these parameters in the students of Birjand University of Medical Science. Materials and Methods: The present study was a descriptive-analytic, cross-sectional type. The study population included all the students at Birjand University of Medical Sciences, selected through stratified randomized sampling method. In order to study IQ, cultural intelligence, and self-monitoring parameters R & B Cattell scale (Scale III, Erli’s Cultural Intelligence Inventory, and Snyder’s Self-monitoring Test were applied, respectively. The obtained data was fed into SPSS (V:21 software using Pearson correlation test, ANOVA, and t-test at the significant level of P≤0.05. Results: From a total of 171 subjects participating in the study, 53.2% were female. The average age of the participants was 21.3±2.7 years. The average IQ, cultural intelligence, and self-monitoring scores were 106±10.44, 85.73±17.31, and 12.35±3.20, respectively. There was a significant correlation between cultural intelligence and self-monitoring (P<0.000; r=0/37. However, there were no significant associations between cultural intelligence and IQ scores as well as between self-monitoring and IQ scores. Conclusion: Regarding the unfavorable cultural intelligence’ skills and abilities ;and their acquirable nature, it is suggested that University consider a significant position for educational and cultural programs in order to enhance cultural intelligence.

  12. Application of near-infrared spectroscopy for monitoring and control of cell culture and fermentation.

    Science.gov (United States)

    Cervera, Albert E; Petersen, Nanna; Lantz, Anna Eliasson; Larsen, Anders; Gernaey, Krist V

    2009-01-01

    Near-infrared (NIR) spectroscopy can potentially provide on-line information on substrate, biomass, product, and metabolite concentrations in fermentation processes, which could be useful for improved monitoring or control. However, several factors can negatively influence the quality of chemometric models built for interpretation of the spectra, thus impairing the analyte concentration predictions. The aim of this review was to provide an overview of necessary conditions and challenges that one has to face when developing a NIR application for monitoring of cell culture or fermentation processes. Important practical aspects are introduced, such as sampling, modeling of biomass concentration, influence of microorganism morphology on the spectra, effects of the hydrodynamic conditions in the fermenter, temperature influence, instrument settings, and signal optimization. Several examples from the literature are provided, which will hopefully guide the reader interested in the topic. Furthermore, the general procedure used for the development of calibration models is presented, and the influence of microorganism metabolism-potential source of correlation between analytes-is commented. Other important issues such as wavelength selection and evaluation of robustness are shortly introduced. Finally, some examples of potential applications of NIR monitoring are provided, including the implementation of control strategies, the combination with other monitoring tools (the so-called sensor fusion), and the description of process trajectories. On the basis of the review, we conclude that acceptance of NIR spectroscopy as a standard monitoring tool by the fermentation industry will necessitate considerably more on-line studies using industrially relevant-and highly challenging-fermentation conditions (high aeration intensity, high biomass concentration and viscosity, and filamentous production strain). (c) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009.

  13. A system for Real time monitoring of buildings with cultural heritage importance using wireless sensor networks

    Science.gov (United States)

    Hloupis, G.; Vallianatos, F.

    2009-04-01

    Simultaneously real time monitoring of buildings usually requires several different sensors. Even if the number of monitoring items is small the cost of monitoring devices plus the telemetry needed could increased in excessive values. For this reason the use of autonomous sensors is indicated. These devices are self contained embedded computers capable of hosting several sensors and communication boards and providing local computing processing. The acquisition of high resolution physical quantities using low power wireless sensor nodes consist a Wireless Sensor Network (WSN). By using these systems it is easy to collect different data from different clusters of sensors using low cost sensor nodes. The main concern for these approaches is the optimization of data acquisition regarding the management of energy capacity and available radio bandwidth. In this study we propose an optimized management scheme for monitoring historical buildings at the city of Chania using sensor nodes connected to high resolution uniaxial and triaxial embedded accelerometers. A number of sensor nodes are placed in every building. Since an event is not a linear process regarding its time occurrence and the produced results in each sensor node (due to sensor temporary malfunction or existence of noise) we followed a non linear approach. The proposed management scheme focus on the optimum self configuration of the network in a hybrid star topology. It is based on public available TinyOS and produces hierarchical rules in order to have at least one central node (the one that sends all the data to the remote data centre). Example policies that demonstrated is thresholding, noise removal, triggering and event correlation which are implemented using wavelet transform techniques. Acknowledgements This work is partially supported by SE-RISK Project (INTERREG III, STRAND B ARCHIMED/Axis: Integrated and Sustainable Management of Cultural and Natural Resources and of Landscape and Risk Management)

  14. A biocompatible micro cell culture chamber (mu CCC) for the culturing and on-line monitoring of eukaryote cells

    DEFF Research Database (Denmark)

    Stangegaard, Michael; Petronis, Sarunas; Jørgensen, Anders Michael

    2006-01-01

    We have previously shown that a polymeric (PMMA) chip with medium perfusion and integrated heat regulation provides sufficiently precise heat regulation, pH-control and medium exchange to support cell growth for weeks. However, it was unclear how closely the cells cultured in the chip resembled c...... compared to cell cultured in culture flasks incubated in a dark and CO2 conditioned incubator....

  15. Non-Invasive Optical Sensor Based Approaches for Monitoring Virus Culture to Minimize BSL3 Laboratory Entry

    Directory of Open Access Journals (Sweden)

    Viswanath Ragupathy

    2015-06-01

    Full Text Available High titers of infectious viruses for vaccine and diagnostic reference panel development are made by infecting susceptible mammalian cells. Laboratory procedures are strictly performed in a Bio-Safety Level-3 (BSL3 laboratory and each entry and exit involves the use of  disposable Personnel Protective Equipment (PPE to observe cell culture conditions. Routine PPE use involves significant recurring costs. Alternative non-invasive optical sensor based approaches to remotely monitor cell culture may provide a promising and cost effective approach to monitor infectious virus cultures resulting in lower disruption and costs. We report here the monitoring of high titer cultures of Human Immunodeficiency Virus-1 (HIV-1 and Herpes Simplex Virus-2 (HSV-2 remotely with the use of optical oxygen sensors aseptically placed inside the cell culture vessel. The replacement of culture media for cell and virus propagation and virus load monitoring was effectively performed using this fluorescent sensor and resulted in half the number of visits to the BSL3 lab (five versus ten.

  16. A biocompatible micro cell culture chamber (mu CCC) for the culturing and on-line monitoring of eukaryote cells

    DEFF Research Database (Denmark)

    Stangegaard, Michael; Petronis, Sarunas; Jørgensen, Anders Michael

    2006-01-01

    on cell survival. Low grade light exposure was however compatible with optical recordings as well as cell viability. These results strongly indicate that a cell culture chip could be constructed that allowed for on-line optical recording of cellular events without affecting the cell culturing condition...... culture chip compared to cell culture flasks. The cell culture chip could without further modification support cell growth of two other cell lines. Light coming from the microscope lamp during optical recordings of the cells was the only external factor identified, that could have a negative effect...

  17. Purification and characterization of three chitinases and one beta-1,3-glucanase accumulating in the medium of cell suspension cultures of barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Kragh, K.M.; Jacobsen, S.; Dalgaard Mikkelsen, J.

    1991-01-01

    -terminal sequence (24 residues) this enzyme was identified as the previously described beta-1,3-endoglucanase GII from germinating barley grain. The results suggest that secretion of chitinase and beta-1,3-glucanase from the embryogenic cell suspensions of barley corresponds to accumulation of the same enzymes...... chromatography. Two of the chitinases were identified as the previously described endochitinases T and C from barley grain. The third and novel chitinase, designated K, was the major basic chitinase in the medium constituting 4% of the soluble protein. Chitinase K was found to be a 33-kDa endochitinase with p......I at 8.7. Further analysis showed that this enzyme is also expressed in barley grain. The amino acid composition and five partial amino acid sequences covering 93 residues of chitinase K were determined. A high similarity was found between chitinase K and barley chitinase T and C as well as basic...

  18. Surveying and Monitoring of Cultural Heritage: The Role of COSMO-SkyMed Mission

    Science.gov (United States)

    Sacco, Patrizia; Battagliere, Maria Libera; Daraio, Maria Girolamo; Coletta, Alessandro

    2016-08-01

    Early applications of SAR in archaeology date back to the 1980s and, although being mainly demonstrative experimentations, they enabled numerous important discoveries. With the development of Synthetic Aperture Radar (SAR) in terms of multi-band, multi- polarization and high-resolution data, space radar remote sensing for archaeology has become a potential field for research. Although the archaeological detection capability of this technology has so far not been fully assessed, the advent of Very High Resolution (VHR) space borne SAR sensors, such as Italian COSMO- SkyMed, offered advanced mapping capability at high resolution for archaeology investigations. This paper aims to give an overview of the contribution provided by the COSMO-SkyMed mission in the framework of specific projects in which SAR X-band data have been exploited for the monitoring and the management of cultural and archeological heritage, showing some obtained results.

  19. Development of an automated chip culture system with integrated on-line monitoring for maturation culture of retinal pigment epithelial cells

    Directory of Open Access Journals (Sweden)

    Mee-Hae Kim

    2017-10-01

    Full Text Available In cell manufacturing, the establishment of a fully automated, microfluidic, cell culture system that can be used for long-term cell cultures, as well as for process optimization is highly desirable. This study reports the development of a novel chip bioreactor system that can be used for automated long-term maturation cultures of retinal pigment epithelial (RPE cells. The system consists of an incubation unit, a medium supply unit, a culture observation unit, and a control unit. In the incubation unit, the chip contains a closed culture vessel (2.5 mm diameter, working volume 9.1 μL, which can be set to 37 °C and 5% CO2, and uses a gas-permeable resin (poly- dimethylsiloxane as the vessel wall. RPE cells were seeded at 5.0 × 104 cells/cm2 and the medium was changed every day by introducing fresh medium using the medium supply unit. Culture solutions were stored either in the refrigerator or the freezer, and fresh medium was prepared before any medium change by warming to 37 °C and mixing. Automated culture was allowed to continue for 30 days to allow maturation of the RPE cells. This chip culture system allows for the long-term, bubble-free, culture of RPE cells, while also being able to observe cells in order to elucidate their cell morphology or show the presence of tight junctions. This culture system, along with an integrated on-line monitoring system, can therefore be applied to long-term cultures of RPE cells, and should contribute to process control in RPE cell manufacturing.

  20. Real-time monitoring of cellular dynamics using a microfluidic cell culture system with integrated electrode array and potentiostat

    DEFF Research Database (Denmark)

    Zor, Kinga; Vergani, M.; Heiskanen, Arto

    2011-01-01

    A versatile microfluidic, multichamber cell culture and analysis system with an integrated electrode array and potentiostat suitable for electrochemical detection and microscopic imaging is presented in this paper. The system, which allows on-line electrode cleaning and modification, was developed...... for real-time monitoring of cellular dynamics, exemplified in this work by monitoring of redox metabolism inside living yeast cells and dopamine release from PC12 cells....

  1. Autologous epidermal cell suspension: A promising treatment for chronic wounds.

    Science.gov (United States)

    Zhao, Hongliang; Chen, Yan; Zhang, Cuiping; Fu, Xiaobing

    2016-02-01

    Chronic wounds have become an increasing medical and economic problem of aging societies because they are difficult to manage. Skin grafting is an important treatment method for chronic wounds, which are refractory to conservative therapy. The technique involving epidermal cell suspensions was invented to enable the possibility of treating larger wounds with only a small piece of donor skin. Both uncultured and cultured autologous epidermal cell suspensions can be prepared and survive permanently on the wound bed. A systematic search was conducted of EMBASE, Cochrane Library, PubMed and web of science by using Boolean search terms, from the establishment of the database until May 31, 2014. The bibliographies of all retrieved articles in English were searched. The search terms were: (epithelial cell suspension OR keratinocyte suspension) and chronic and wound. From the included, 6 studies are descriptive interventions and discussed the use of autologous keratinocyte suspension to treat 61 patients' chronic wound. The various methods of preparation of epidermal cell suspension are described. The advantages and shortcomings of different carriers for epidermal cell suspensions are also summarised. Both uncultured and cultured autologous epidermal cell suspensions have been used to treat chronic wounds. Although the limitations of these studies include the small number of patient populations with chronic wounds and many important problems that remain to be solved, autologous epidermal cell suspension is a promising treatment for chronic wounds. Copyright © 2015 Tissue Viability Society. Published by Elsevier Ltd. All rights reserved.

  2. Design of a Basic System of Indicators for Monitoring and Evaluating Spanish Cooperation's Culture and Development Strategy

    Science.gov (United States)

    Coll-Serrano, Vicente; Carrasco-Arroyo, Salvador; Blasco-Blasco, Olga; Vila-Lladosa, Luis

    2012-01-01

    Objective: This article describes the process implemented in order to define and build up a Basic Monitoring and Evaluation System for Spanish Cooperation's Culture and Development Strategy (CD-S). Research Design: Delphi techniques were used to assess a wide catalogue of indicators for each of the strategic areas included in the CD-S. Afterward,…

  3. Promising Data for Public Empowerment: The Making of Data Culture and Water Monitoring Infrastructures in the Marcellus Shale Gas Rush

    Science.gov (United States)

    Jalbert, Kirk

    A recent wave of advanced technologies for collecting and interpreting data offer new opportunities for laypeople to contribute to environmental monitoring science. This dissertation examines the conditions in which building knowledge infrastructures and embracing data "cultures" empowers and disempowers communities to challenge polluting industries. The processes and technologies of data cultures give people new capacities to understand their world, and to formulate powerful scientific arguments. However, data cultures also make many aspects of social life invisible, and elevate quantitative objective analysis over situated, subjective observation. This study finds that data cultures can empower communities when concerned citizens are equal contributors to research partnerships; ones that enable them to advocate for more nuanced data cultures permitting of structural critiques of status-quo environmental governance. These arguments are developed through an ethnographic study of participatory watershed monitoring projects that seek to document the impacts of shale gas extraction in Pennsylvania, New York, and West Virginia. Energy companies are drilling for natural gas using highly controversial methods of extraction known as hydraulic fracturing. Growing evidence suggests that nearby watersheds can be impacted by a myriad of extraction related problems including seepage from damaged gas well casing, improper waste disposal, trucking accidents, and the underground migration of hydraulic fracking fluids. In response to these risks, numerous organizations are coordinating and carrying out participatory water monitoring efforts. All of these projects embrace data culture in different ways. Each monitoring project has furthermore constructed its own unique infrastructure to support the sharing, aggregation, and analysis of environmental data. Differences in data culture investments and infrastructure building make some projects more effective than others in empowering

  4. [Effect of manganese (II), cobalt (II), and nickel (II) ions on the growth and production of coumarins in the suspension culture of Angelica archangelica L].

    Science.gov (United States)

    Siatka, T; Kasparová, M; Sklenárová, H; Solich, P

    2005-01-01

    The plant cell reacts to an increased concentration of metals in the environment by various mechanisms. They include an increase in the formation of heat-shock proteins, metallothioneins, phytochelatins, amino acids (cysteine, histidine), organic acids (citric, malic), or secondary metabolites. The latter mechanism is being investigated for its possible use in explant cultures for the stimulation of secondary metabolism, which is the source of substances of pharmaceutical importance. The study tested manganese (II) (0, 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, and 50 mM in the medium), cobalt (II), and nickel (II) ions (0, 0.1, 0.5, 1, 5, 10, 50, 100, 200, and 500 microM in the medium) as potential elicitors of coumarin production. At the same time, toxicity of these metals for the culture was examined by evaluating their effect on growth (characterized by fresh and dry weight of biomass at the end of a two-week cultivation). Cultures were cultivated in the dark and in the light. It has been found that the growth of cultures is not influenced by manganese in concentrations ranging from 0 to 2 mM, then it slightly decreases, at a concentration of 50 mM it is lower by 20 % when cultivated in the dark and by 30 % when cultivated in the light in comparison with the control. Cobalt in concentrations of 0 to 50 microM does not significantly influence the growth of the culture, higher concentrations decrease the biomass yields, more markedly when cultivated in the light (at 500 microM Co by 60 %, in the dark only by 30 % in comparison with the controls). Nickel in concentrations of 0.1 to 200 microM does not influence growth, and in a concentration of 500 microM decreases it by approximately 30 % in comparison with the control both in the light and dark. Production of coumarins was not stimulated by any metal in comparison with the control cultures, only the removal of manganese from the medium in the culture cultivated in the dark increased production by about 15 % versus the

  5. A service dedicated to Cultural Heritage Risk Assessment and Monitoring on the Web

    Science.gov (United States)

    Dore, Nicole; Monteleone, Antonio; Benenati, Luca; Bernardi, Lorenzo; Giovagnoli, Annamaria; Cacace, Carlo

    2017-04-01

    VIDEOR project, financed by the Italian Ministry of Economic Development (MISE) and strongly supported by the Ministry of Cultural Heritage (MiBACT), is developed by NAIS (Nextant Applications and Innovative Solutions) in collaboration with ISCR (Institute for Conservation and Restoration, MiBACT) and SUPERELECTRIC s.r.l. The project has the aim to provide a service to public institutions responsible of CH preservation, maintenance and restoration, for the assessment of the potential level of aggressiveness of factors responsible for cultural heritage degradation. VIDEOR represents the first example of a continuative monitoring, consultable on the web and constantly updated. VIDEOR is based on the production of a set of products that will help institutions in the evaluation of threats linked to damages and/or loss of the cultural asset. This new approach of cultural heritage condition assessment will support "Carta del Rischio" Italian methodology, a GIS for a scientific and administrative support furnished to Public Entities and developed by ISCR. Test site selected for project demonstration is the archaeological area of Villa Adriana, UNESCO site since 1999. The property, located near Tivoli town (30 km east from Rome), has an extension of 80ha and the buffer zone has an extension of 500ha. This area, near Tivoli and not far from Rome -political and administrative location of the Roman Empire- was chosen by Adriano emperor for the construction of his magnificent residence. VIDEOR products and analyses are based on data coming from several sensors, such as satellites images (optical and SAR) and drones, these last used when satellites spatial resolution is considered not appropriate or when, after severe events, deeper evaluations are necessary. After the earthquake swarm that interested Italy from August 2016 to January 2017 and that destroyed a huge amount of unmovable cultural properties close to zone of the epicenter, analyses were performed over the test site

  6. Micro fluidic System for Culturing and Monitoring of Neuronal Cells and Tissue

    DEFF Research Database (Denmark)

    Bakmand, Tanya; Waagepetersen, Helle S.

    The aim of this Ph.D. project was to combine experience within cell and tissue culturing, electrochemistry and microfabrication in order to develop an in vivo-like fluidic culturing platform, challenging the traditional culturing methods. The first goal was to develope a fluidic system for cultur...

  7. Routing Topologies of Wireless Sensor Networks for Health Monitoring of a Cultural Heritage Site.

    Science.gov (United States)

    Aparicio, Sofía; Martínez-Garrido, María I; Ranz, Javier; Fort, Rafael; Izquierdo, Miguel Ángel G

    2016-10-19

    This paper provides a performance evaluation of tree and mesh routing topologies of wireless sensor networks (WSNs) in a cultural heritage site. The historical site selected was San Juan Bautista church in Talamanca de Jarama (Madrid, Spain). We report the preliminary analysis required to study the effects of heating in this historical location using WSNs to monitor the temperature and humidity conditions during periods of weeks. To test which routing topology was better for this kind of application, the WSNs were first deployed on the upper floor of the CAEND institute in Arganda del Rey simulating the church deployment, but in the former scenario there was no direct line of sight between the WSN elements. Two parameters were selected to evaluate the performance of the routing topologies of WSNs: the percentage of received messages and the lifetime of the wireless sensor network. To analyze in more detail which topology gave the best performance, other communication parameters were also measured. The tree topology used was the collection tree protocol and the mesh topology was the XMESH provided by MEMSIC (Andover, MA, USA). For the scenarios presented in this paper, it can be concluded that the tree topology lost fewer messages than the mesh topology.

  8. Routing Topologies of Wireless Sensor Networks for Health Monitoring of a Cultural Heritage Site

    Directory of Open Access Journals (Sweden)

    Sofía Aparicio

    2016-10-01

    Full Text Available This paper provides a performance evaluation of tree and mesh routing topologies of wireless sensor networks (WSNs in a cultural heritage site. The historical site selected was San Juan Bautista church in Talamanca de Jarama (Madrid, Spain. We report the preliminary analysis required to study the effects of heating in this historical location using WSNs to monitor the temperature and humidity conditions during periods of weeks. To test which routing topology was better for this kind of application, the WSNs were first deployed on the upper floor of the CAEND institute in Arganda del Rey simulating the church deployment, but in the former scenario there was no direct line of sight between the WSN elements. Two parameters were selected to evaluate the performance of the routing topologies of WSNs: the percentage of received messages and the lifetime of the wireless sensor network. To analyze in more detail which topology gave the best performance, other communication parameters were also measured. The tree topology used was the collection tree protocol and the mesh topology was the XMESH provided by MEMSIC (Andover, MA, USA. For the scenarios presented in this paper, it can be concluded that the tree topology lost fewer messages than the mesh topology.

  9. Advanced process monitoring and feedback control to enhance cell culture process production and robustness.

    Science.gov (United States)

    Zhang, An; Tsang, Valerie Liu; Moore, Brandon; Shen, Vivian; Huang, Yao-Ming; Kshirsagar, Rashmi; Ryll, Thomas

    2015-12-01

    It is a common practice in biotherapeutic manufacturing to define a fixed-volume feed strategy for nutrient feeds, based on historical cell demand. However, once the feed volumes are defined, they are inflexible to batch-to-batch variations in cell growth and physiology and can lead to inconsistent productivity and product quality. In an effort to control critical quality attributes and to apply process analytical technology (PAT), a fully automated cell culture feedback control system has been explored in three different applications. The first study illustrates that frequent monitoring and automatically controlling the complex feed based on a surrogate (glutamate) level improved protein production. More importantly, the resulting feed strategy was translated into a manufacturing-friendly manual feed strategy without impact on product quality. The second study demonstrates the improved process robustness of an automated feed strategy based on online bio-capacitance measurements for cell growth. In the third study, glucose and lactate concentrations were measured online and were used to automatically control the glucose feed, which in turn changed lactate metabolism. These studies suggest that the auto-feedback control system has the potential to significantly increase productivity and improve robustness in manufacturing, with the goal of ensuring process performance and product quality consistency. © 2015 Wiley Periodicals, Inc.

  10. Sucrose metabolizing enzymes in cell suspension cultures of Bauhinia forficata, Curcuma zedoaria and Phaseolus vulgaris Enzimas do metabolismo da sacarose em cultura celular de Bauhinia forficata, Curcuma zedoaria e Phaseolus vulgaris

    Directory of Open Access Journals (Sweden)

    Marcia Ometto de Mello

    2001-09-01

    Full Text Available The objective of this work was to study the activity of sucrose metabolizing enzymes in extracts of cell suspension cultures of Bauhinia forficata Link, Curcuma zedoaria Roscoe and Phaseolus vulgaris L. Invertase pathway was identified in the three studied species. Sucrose synthase pathway was also responsible for sucrose metabolism in Curcuma zedoaria and Phaseolus vulgaris cells. Activity values higher than 300 nmol min-1 mg-1 of protein were found for acid and neutral invertases, UDPglucose pyrophosphorylase and phosphoglucomutase in the cell extract of the three plant species. Sucrose synthase showed low activity in Bauhinia forficata cells. As sucrose concentration in the culture medium decreased, sucrose synthase activity increased in C. zedoaria and P. vulgaris cells. The glycolytic enzymes activity gradually reduced at the end of the culture period, when carbohydrate was limited.O objetivo deste trabalho foi estudar as enzimas do metabolismo da sacarose em culturas de célula em suspensão de Bauhinia forficata Link, Curcuma zedoaria Roscoe e Phaseolus vulgaris L. A via da invertase foi identificada nas três espécies estudadas. A via da sacarose sintase também foi responsável pelo metabolismo da sacarose em células de Curcuma zedoaria e Phaseolus vulgaris. Foram encontradas atividades maiores que 300 nmol min-1 mg-1 de proteína das enzimas invertase ácida e alcalina, UDPglicose pirofosforilase e fosfoglicomutase no extrato celular das três espécies de plantas. A sacarose sintase mostrou atividade baixa nas células de Bauhinia forficata. À medida que a concentração de sacarose no meio de cultura diminuiu, a atividade da sacarose sintase aumentou em células de Curcuma zedoaria e Phaseolus vulgaris. Ao final do período de cultura, quando os carboidratos se tornaram limitantes, as atividades das enzimas glicolíticas reduziram-se gradualmente.

  11. Suture midface suspension

    Directory of Open Access Journals (Sweden)

    Murthy Rachna

    2006-11-01

    Full Text Available Abstract Objective To describe a simple and effective facelift technique useful as an adjunct to other oculoplastic procedures Methods Retrospective, non-comparative case series. Thirty five patients undergoing suture midface suspension from 1998 to 2000. Suspension sutures were passed from the nasolabial fold to the temporalis fascia to elevate the midface and the corner of the mouth. Results A satisfactory and stable outcome is obtained in 2 years of follow up. Conclusion Suture midface suspension is a safe and effective technique for the management of midface descent.

  12. Numerical and clinical precision of continuous glucose monitoring in Colombian patients treated with insulin infusion pump with automated suspension in hypoglycemia.

    Science.gov (United States)

    Gómez, Ana M; Marín Sánchez, Alejandro; Muñoz, Oscar M; Colón Peña, Christian Alejandro

    2015-12-01

    Insulin pump therapy associated with continuous glucose monitoring has shown a positive clinical impact on diabetes control and reduction of hypoglycemia episodes. There are descriptions of the performance of this device in other populations, but its precision and accuracy in Colombia and Latin America are unknown, especially in the routine outpatient setting. Data from 33 type 1 and type 2 diabetes patients with sensor-augmented pump therapy with threshold suspend automation, MiniMed Paradigm® Veo™ (Medtronic, Northridge, California), managed at Hospital Universitario San Ignacio (Bogotá, Colombia) and receiving outpatient treatment, were analyzed. Simultaneous data from continuous glucose monitoring and capillary blood glucose were compared, and their precision and accuracy were calculating with different methods, including Clarke error grid. Analyses included 2,262 continuous glucose monitoring -reference paired glucose values. A mean absolute relative difference of 20.1% was found for all measurements, with a value higher than 23% for glucose levels ≤75mg/dL. Global compliance with the ISO criteria was 64.9%. It was higher for values >75mg/dl (68.3%, 1,308 of 1,916 readings), than for those ≤ 75mg/dl (49.4%, 171 of 346 readings). Clinical accuracy, as assessed by the Clarke error grid, showed that 91.77% of data were within the A and B zones (75.6% in hypoglycemia). A good numerical accuracy was found for continuous glucose monitoring in normo and hyperglycemia situations, with low precision in hypoglycemia. The clinical accuracy of the device was adequate, with no significant safety concerns for patients. Copyright © 2015 SEEN. Published by Elsevier España, S.L.U. All rights reserved.

  13. Induced accumulation of 20-hydroxyecdysone in cell suspension ...

    African Journals Online (AJOL)

    This study describes the effects of culture medium, culture temperature, sucrose concentration and cholesterol feeding on cell growth and 20-hydroxyecdysone production in suspension cultures of Vitex glabrata, an important medicinal plant in Thailand. Cell growth and 20-hydroxyecdysone production were not significantly ...

  14. Induced accumulation of 20-hydroxyecdysone in cell suspension ...

    African Journals Online (AJOL)

    Administrator

    2011-09-12

    Sep 12, 2011 ... This study describes the effects of culture medium, culture temperature, sucrose concentration and cholesterol feeding on cell growth and 20-hydroxyecdysone production in suspension cultures of Vitex glabrata, an important medicinal plant in Thailand. Cell growth and 20-hydroxyecdysone production.

  15. In vitro morphogenesis and cell suspension culture establishment in Piper solmsianum C. DC. (Piperaceae Morfogênese in vitro e estabelecimento de culturas de suspensão celular em Piper solmsianum C. DC. (Piperaceae

    Directory of Open Access Journals (Sweden)

    Tiago Santana Balbuena

    2009-03-01

    Full Text Available Piper solmsianum is a shrub from Southeast Brazil in which many biologically active compounds were identified. The aim of this work was to establish a cell suspension culture system for this species. With this in mind, petiole and leaf explants obtained from in vitro plantlets were cultured in the presence of different plant growth regulator combinations (IAA, NAA, 2,4-D and BA. Root and indirect shoot adventitious formation, detected by histological analysis, was observed. Besides the different combinations of plant growth regulators, light regime and the supplement of activated charcoal (1.5 mg.l-1 were tested for callus induction and growth. Cultures maintained in light, on a 0.2 mg.l-1 2,4-D and 2 mg.l-1 BA supplemented medium, and in the absence of activated charcoal, showed the highest calli fresh matter increment. From a callus culture, cell suspension cultures were established and their growth and metabolite accumulation studied. The achieved results may be useful for further characterization of the activated secondary metabolites pathways in in vitro systems of P. solmsianum.Piper solmsianum é uma espécie herbácea do sudeste brasileiro onde vários compostos biologicamente ativos já foram identificados. O objetivo deste trabalho foi estabelecer suspensões celulares nesta espécie. Para tanto, foram utilizados explantes de pecíolos e folhas, retirados de plântulas cultivadas in vitro, os quais foram submetidos a diferentes combinações de reguladores de crescimento (AIA, ANA, 2,4-D e BAP. Foi obtida a neo-formação de raízes e brotos, estes últimos através do processo de organogênese indireta evidenciada por estudos histológicos. Para a indução e crescimento dos calos, foram avaliados, além das diferentes combinações de reguladores de crescimento, a suplementação ao meio de cultura de carvão ativado (1,5 mg.l-1 e o regime de luz. Culturas mantidas na luz, em meio de cultura suplementado com 0,2 mg.l-1 2,4-D e 2 mg

  16. The Effect of Multicultural Experience in Conflicts Management Styles: Mediation of Cultural Intelligence and Self-Monitoring

    Directory of Open Access Journals (Sweden)

    Gabriela Gonçalves

    2015-03-01

    Full Text Available Conflict is an inevitable reality both in personal and in organizational life. For being inevitable, the conflict must be managed Defined as a process that occurs when one party feels adversely affected by another (e.g., De Dreu, 1997 the conflict management styles can be analysed as a function of personality variables. In this respect the cultural intelligence, self-monitoring and self-interdependent seem to be relevant variables, since characterised by flexibility and interest in other aspects present in conflict management styles. In this study, we propose that cultural intelligence, associated with the self-interdependent and self-monitoring, can have a positive impact on the choice of most effective interpersonal conflict resolution styles. Being cultural intelligence an attribute of extreme importance, we still sought to determine how the quantity and quality of intercultural contact and self-interdependent present themselves as predictors of it. With a sample of 399 individuals, the proposed model suggests that high levels of cultural intelligence mediated by a high self-monitoring and selfinterdependent positively affect and predict the conflict resolution styles adopted. Given the need to develop abilities aimed at increasing the skills of conflict resolution, this study adds to the existing literature new predictors, contributing to the welfare and performance of human resources, and consequently to success and organizational effectiveness.

  17. Innovative systems for cultural heritage conservation. Millimeter wave application for non-invasive monitoring and treatment of works of art.

    Science.gov (United States)

    Bruno, Bisceglia; De Leo, Roberto; Pastore, Anna Pia; von Gratowski, Svetlana; Meriakri, Viatcheslav

    2011-01-01

    A novel non invasive technique and a suitable apparatus for disinfestation of artworks is introduced. Non destructive and non invasive techniques are often irreplaceable in order to preserve and restore cultural heritage objects in its structure and shape. Although many techniques are available for art and archaeological works the non invasive methods are preferred as they leave the object untouched after treatment. Environmental parameters, such as humidity, can damage culture heritage objects and also results in spring up variety of pests and other micro-organisms. Non-invasive monitoring of these damage and also disinfestation treatments and drying with help of electromagnetic waves are preferred as they keep the object untouched after treatment. Application of millimeter waves for solving this problem is discussed here. Millimeter waves have high spatial resolution and absorption in water as well as in bio-objects that are usually moist and at the same time minimal interaction with dry culture heritage objects by itself. Different phases of the microwaves treatment (MW) of artworks are described, some results are shown and discussed. Many biological forms don't survive over a certain temperature, called lethal temperature which, for most xylophages is about 53-55 degrees C, while for moulds and funguses is between 65 and 70 degrees C. In order to evaluate the management of disinfestation of works of art, incident power, temperature, exposure time were monitored. The monitoring of temperature is essential in order to prevent damages. A computer simulation allows to predict and monitor the heating process.

  18. Design of a hybrid (wired/wireless) acquisition data system for monitoring of cultural heritage physical parameters in Smart Cities.

    Science.gov (United States)

    García Diego, Fernando-Juan; Esteban, Borja; Merello, Paloma

    2015-03-25

    Preventive conservation represents a working method and combination of techniques which helps in determining and controlling the deterioration process of cultural heritage in order to take the necessary actions before it occurs. It is acknowledged as important, both in terms of preserving and also reducing the cost of future conservation measures. Therefore, long-term monitoring of physical parameters influencing cultural heritage is necessary. In the context of Smart Cities, monitoring of cultural heritage is of interest in order to perform future comparative studies and load information into the cloud that will be useful for the conservation of other heritage sites. In this paper the development of an economical and appropriate acquisition data system combining wired and wireless communication, as well as third party hardware for increased versatility, is presented. The device allows monitoring a complex network of points with high sampling frequency, with wired sensors in a 1-wire bus and a wireless centralized system recording data for monitoring of physical parameters, as well as the future possibility of attaching an alarm system or sending data over the Internet. This has been possible with the development of three board's designs and more than 5000 algorithm lines. System tests have shown an adequate system operation.

  19. Design of a Hybrid (Wired/Wireless Acquisition Data System for Monitoring of Cultural Heritage Physical Parameters in Smart Cities

    Directory of Open Access Journals (Sweden)

    Fernando-Juan García Diego

    2015-03-01

    Full Text Available Preventive conservation represents a working method and combination of techniques which helps in determining and controlling the deterioration process of cultural heritage in order to take the necessary actions before it occurs. It is acknowledged as important, both in terms of preserving and also reducing the cost of future conservation measures. Therefore, long-term monitoring of physical parameters influencing cultural heritage is necessary. In the context of Smart Cities, monitoring of cultural heritage is of interest in order to perform future comparative studies and load information into the cloud that will be useful for the conservation of other heritage sites. In this paper the development of an economical and appropriate acquisition data system combining wired and wireless communication, as well as third party hardware for increased versatility, is presented. The device allows monitoring a complex network of points with high sampling frequency, with wired sensors in a 1-wire bus and a wireless centralized system recording data for monitoring of physical parameters, as well as the future possibility of attaching an alarm system or sending data over the Internet. This has been possible with the development of three board’s designs and more than 5000 algorithm lines. System tests have shown an adequate system operation.

  20. Urinary incontinence - retropubic suspension

    Science.gov (United States)

    ... your doctor will have you try bladder retraining, Kegel exercises, medicines, or other options. If you tried ... retropubic colposuspension; Needle suspension; Burch colposuspension Patient Instructions Kegel exercises - self-care Self catheterization - female Suprapubic catheter ...

  1. Rheology of organoclay suspension

    CSIR Research Space (South Africa)

    Hato, MJ

    2011-05-01

    Full Text Available The authors have studied the rheological properties of clay suspensions in silicone oil, where clay surfaces were modified with three different types of surfactants. Dynamic oscillation measurements showed a plateau-like behavior for all...

  2. Identification of Alternaria alternata Mycotoxins by LC-SPE-NMR and Their Cytotoxic Effects to Soybean (Glycine max Cell Suspension Culture

    Directory of Open Access Journals (Sweden)

    Edson Rodrigues-Filho

    2013-02-01

    Full Text Available This present work describes the application of liquid chromatograpy-solid phase extraction-nuclear magnetic resonance spectroscopy to analyse Alternaria alternata crude extracts. Altenusin (1, alternariol (2, 3'-hydroxyalternariol monomethyl ether (3, and alternariol monomethyl ether (4, were separated and identified. High-resolution mass spectrometry confirmed the proposed structures. The cytotoxic effects of these compounds towards plants were determined using soybean (Glycine max cell cultures as a model. EC50 values which range from 0.11 (±0.02 to 4.69 (±0.47 μM showed the high cytotoxicity of these compounds.

  3. The Mystical Suspension

    Directory of Open Access Journals (Sweden)

    Héctor Santiesteban Oliva

    2016-11-01

    Full Text Available Mistical suspension, silence, time, absolute, ontology, ineffability, aletheiaIn the mystical ecstasy there is a sensorial and intellectual suspension when contemplating the absolute, the ontological Being. Silence is not only significant: it is revealing. The greatest expression of experience inner silence . The word is insufficient when the ontological reality is revealed. Revelation or truth , the Greek concept of aletheia, takes on greater significance in that transcendental experience. It is also suspended phenomenological time and remains eternity open.

  4. Predictive modeling in Clostridium acetobutylicum fermentations employing Raman spectroscopy and multivariate data analysis for real-time culture monitoring

    Science.gov (United States)

    Zu, Theresah N. K.; Liu, Sanchao; Germane, Katherine L.; Servinsky, Matthew D.; Gerlach, Elliot S.; Mackie, David M.; Sund, Christian J.

    2016-05-01

    The coupling of optical fibers with Raman instrumentation has proven to be effective for real-time monitoring of chemical reactions and fermentations when combined with multivariate statistical data analysis. Raman spectroscopy is relatively fast, with little interference from the water peak present in fermentation media. Medical research has explored this technique for analysis of mammalian cultures for potential diagnosis of some cancers. Other organisms studied via this route include Escherichia coli, Saccharomyces cerevisiae, and some Bacillus sp., though very little work has been performed on Clostridium acetobutylicum cultures. C. acetobutylicum is a gram-positive anaerobic bacterium, which is highly sought after due to its ability to use a broad spectrum of substrates and produce useful byproducts through the well-known Acetone-Butanol-Ethanol (ABE) fermentation. In this work, real-time Raman data was acquired from C. acetobutylicum cultures grown on glucose. Samples were collected concurrently for comparative off-line product analysis. Partial-least squares (PLS) models were built both for agitated cultures and for static cultures from both datasets. Media components and metabolites monitored include glucose, butyric acid, acetic acid, and butanol. Models were cross-validated with independent datasets. Experiments with agitation were more favorable for modeling with goodness of fit (QY) values of 0.99 and goodness of prediction (Q2Y) values of 0.98. Static experiments did not model as well as agitated experiments. Raman results showed the static experiments were chaotic, especially during and shortly after manual sampling.

  5. Chlorogenic acid biosynthesis: characterization of a light-induced microsomal 5-O-(4-coumaroyl)-D-quinate/shikimate 3'-hydroxylase from carrot (Daucus carota L. ) cell suspension cultures

    Energy Technology Data Exchange (ETDEWEB)

    Kuehnl, T.K.; Koch, U.; Heller, W.; Wellmann, E.

    1987-10-01

    Microsomal preparations from carrot (Daucus carota L.) cell suspension cultures catalyze the formation of trans-5-O-caffeoyl-D-quinate (chlorogenate) from trans-5-O-(4-coumaroyl)-D-quinate. trans-5-O-(4-Coumaroyl)shikimate is converted to about the same extent to trans-5-O-caffeoylshikimate. trans-4-O-(4-Coumaroyl)-D-quinate, trans-3-O-(4-coumaroyl)-D-quinate, trans-4-coumarate, and cis-5-O-(4-coumaroyl)-D-quinate do not act as substrates. The reaction is strictly dependent on molecular oxygen and on NADPH as reducing cofactor. NADH and ascorbic acid cannot substitute for NADPH. Cytochrome c, Tetcyclacis, and carbon monoxide inhibit the reaction suggesting a cytochrome P-450-dependent mixed-function monooxygenase. Competition experiments as well as induction and inhibition phenomena indicate that there is only one enzyme species which is responsible for the hydroxylation of the 5-O-(4-coumaric) esters of both D-quinate and shikimate. The activity of this enzyme is greatly increased by in vivo irradiation of the cells with blue/uv light. We conclude that the biosynthesis of the predominant caffeic acid conjugates in carrot cells occurs via the corresponding 4-coumaric acid esters. Thus, in this system, 5-O-(4-coumaroyl)-D-quinate can be seen as the final intermediate in the chlorogenic acid pathway.

  6. The influence of glutamine on growth and viability of cell suspension cultures of Douglas-fir after exposure to polyethylene glycol.

    Science.gov (United States)

    Leustek, T; Kirby, E G

    1988-12-01

    The response of cell cultures of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) to osmotic stress was studied by measuring cell growth and viability after exposure to polyethylene glycol (PEG) (M(r) 6000-8000). Growth of cells inoculated in a medium containing 10% PEG was slightly inhibited, whereas growth in a medium containing 15% PEG was severely inhibited. Cells grown for 6 days in nutrient medium and then subcultured in a medium containing 15% PEG to induce water stress showed high viabilities, whereas cells grown for longer than 6 days before exposure to PEG showed decreased viabilities after subculture. Cells grown in medium containing 30 mM glutamine were significantly more resistant to PEG-induced water stress, as measured by viability, than cells grown in medium without glutamine.

  7. Design of a basic system of indicators for monitoring and evaluating Spanish Cooperation's Culture and Development Strategy.

    Science.gov (United States)

    Coll-Serrano, Vicente; Carrasco-Arroyo, Salvador; Blasco-Blasco, Olga; Vila-Lladosa, Luis

    2012-08-01

    This article describes the process implemented in order to define and build up a Basic Monitoring and Evaluation System for Spanish Cooperation's Culture and Development Strategy (CD-S). Delphi techniques were used to assess a wide catalogue of indicators for each of the strategic areas included in the CD-S. Afterward, those indicators considered most suitable for monitoring and evaluating purposes were selected based on the assessments provided by a panel of experts. As a consequence, the resulting system, made up by a total of 80 indicators, was designed to be a practical, manageable, and targeted tool for its potential users, who include managers and technical officers who map out strategy together with stakeholders involved in its implementation, and specifically with staff at cultural centers abroad and in technical aid offices.

  8. An improved culturing method for opiine fruit fly parasitoids and its application to parasitoid monitoring in the field.

    Science.gov (United States)

    Masry, Ayad; Furlong, Michael J; Clarke, Anthony R; Cunningham, John Paul

    2018-02-01

    Good culturing methods play an important role in the study of insect behavior and its application to pest management. Here, we describe and validate a new method for rearing the parasitoid wasp, Diachasmimorpha kraussii, which attacks some of the world's worst fruit fly pests and is an internationally used biological control agent. Our method differs from standard culturing approaches by presenting adult wasps with host-infested artificial media within a "culturing bag," which mimics a natural (fruit) oviposition substrate. In laboratory trials using wild collected D. kraussii, the culturing bag method was compared to the use of host-infested nectarines, and a commonly used laboratory method of presenting host-infested artificial media within Petri dishes. The culturing bag method proved to be a significant improvement on both methods, combining the advantages of high host survival in artificial media with parasitism levels that were the equivalent to those recorded using host-infested fruits. In our field study, culturing bags infested with the Queensland fruit fly, Bactrocera tryoni, and hung in a mixed peach and nectarine orchard proved to be effective "artificial fruits" attracting wild D. kraussii for oviposition. Significantly more adult wasps were reared from the culturing bags compared to field collected fruits. This was shown to be due to higher fruit fly larval density in the bags, as similar percentage parasitism rates were found between the culturing bags and ripe fruits. We discuss how this cheap, time-efficient method could be applied to collecting and monitoring wild D. kraussii populations in orchards, and assist in maintaining genetic variability in parasitoid laboratory cultures. © 2016 Institute of Zoology, Chinese Academy of Sciences.

  9. Visualization of morphological categories of colonies for monitoring of effect on induced pluripotent stem cell culture status

    Directory of Open Access Journals (Sweden)

    Risako Nagasaka

    2017-06-01

    Full Text Available From the recent advances, there are growing expectations toward the mass production of induced pluripotent stem cells (iPSCs for varieties of applications. For such type of industrial cell manufacturing, the technology which can stabilize the production efficiency is strongly required. Since the present iPSC culture is covered by delicate manual operations, there are still quality differences in produced cells from same culture protocols. To monitor the culture process of iPSCs with the quantified data to evaluate the culture status, we here introduce image-based visualization method of morphological diversity of iPSC colonies. We have set three types of experiments to evaluate the influential factors in iPSC culture technique that may disturb the undifferentiation status of iPSC colonies: (Exp. 1 technical differences in passage skills, (Exp. 2 technical differences in feeder cell preparation, and (Exp. 3 technical differences in maintenance skills (medium exchange frequency with the combination of manual removal of morphologically irregular colonies. By measuring the all existing colonies from real-time microscopic images, the heterogenous change of colony morphologies in the culture vessel was visualized. By such visualization with morphologically categorized Manhattan chart, the difference between technical skills could be compared for evaluating appropriate cell processing.

  10. Effects of gamma irradiation on the plasma membrane of suspension-cultured apple cells. Rapid irreversible inhibition of H+-ATPase activity

    International Nuclear Information System (INIS)

    Dong, C.-Z.; Montillet, J.-L.; Triantaphylides, C.

    1994-01-01

    The effects of ionizing radiation, used in post-harvest treatment of fruit and vegetables. were investigated on cultured apple cells (Pyrus malus L. cv. Royal Red) on a short-term period. Irradiation (2 kGy) induced an increase of passive ion effluxes from cells and a decrease of cell capacity to regulate external pH. These alterations are likely due to effects on plasma membrane structure and function and were further investigated by studying the effects of irradiation on plasma membrane H + -ATPase activity. Plasma membrane-enriched vesicles were prepared and the H + -ATPase activity was characterized. Irradiation of the vesicles induced a dose dependent inhibition of H + -ATPase activity. The loss of enzyme activity was immediate, even at low doses (0.5 kGy), and was not reversed by the addition of 2mM dithiothreitol. This inhibition may be the result of an irreversible oxidation of enzyme sulfhydryl moieties and/or the result of changes induced within the lipid bilayer affecting the membrane-enzyme interactions. Further analysis of the H + -ATPase activity was carried out on vesicles obtained from irradiated cells confirming the previous results. In vivo recovery of activity was not observed within 5 h following the treatment, thus explaining the decrease of cell capacity to regulate external pH. This rapid irreversible inhibition of the plasma membrane H + -ATPase must be considered as one of the most important primary biochemical events occurring in irradiated plant material. (author)

  11. Intracellular salicylic acid is involved in signal cascade regulating low ammonium-induced taxoid biosynthesis in suspension cultures of Taxus chinensis.

    Science.gov (United States)

    Zhou, Xin; Zhong, Jian-Jiang

    2011-05-01

    It was previously reported that low initial ammonium (2 mM) in medium had significant stimulating effects on the biosynthesis of taxuyunnanine C (Tc) by Taxus chinensis cells. However, the secondary metabolism induction mechanism of the low initial ammonium is yet unknown in plant cells. To provide an insight into the defense signals response to the low initial ammonium, oxidative burst and intracellular salicylic acid (SA) were detected, and their influences on the expression of important genes in taxoid biosynthetic pathway were examined in the cell cultures of T. chinensis. Induced H(2)O(2) production, elevated phenylalanine ammonia-lyase (PAL) activity, and enhanced SA biosynthesis were observed. Interestingly, inhibition of SA biosynthesis by paclobutrazol and (BOC-aminooxy) acetic acid significantly depressed the Tc stimulation and up-regulation of Tc biosynthetic genes of geranylgeranyl diphosphate synthase and taxadiene synthase. The role of intracellular SA in regulating Tc biosynthesis was further confirmed by applying exogenous SA in normal ammonium (20 mM) medium. The results indicated that SA acted as a signal in low initial ammonium-induced Tc biosynthesis. A signal transduction cascade from defense signal response to activated transcription of taxoid biosynthetic genes and enhanced Tc production is proposed.

  12. Connecting World Heritage Nominations and Monitoring with the Support of the Silk Roads Cultural Heritage Resource Information System

    Science.gov (United States)

    Vileikis, O.; Dumont, B.; Serruys, E.; Van Balen, K.; Tigny, V.; De Maeyer, P.

    2013-07-01

    Serial transnational World Heritage nominations are challenging the way cultural heritage has been managed and evaluated in the past. Serial transnational World Heritage nominations are unique in that they consist of multiple sites listed as one property, distributed in different countries, involving a large diversity of stakeholders in the process. As a result, there is a need for precise baseline information for monitoring, reporting and decision making. This type of nomination requires different methodologies and tools to improve the monitoring cycle from the beginning of the nomination towards the periodic reporting. The case study of the Silk Roads Cultural Heritage Resource Information System (CHRIS) illustrates the use of a Geographical Content Management System (Geo-CMS) supporting the serial transnational World Heritage nomination and the monitoring of the Silk Roads in the five Central Asian countries. The Silk Roads CHRIS is an initiative supported by UNESCO World Heritage Centre (WHC) and the Belgian Federal Science Policy Office (BELSPO), and developed by a consortium headed by the Raymond Lemaire International Centre for Conservation (RLICC) at the KULeuven. The Silk Roads CHRIS has been successfully assisting in the preparation of the nomination dossiers of the Republics of Kazakhstan, Tajikistan and Uzbekistan and will be used as a tool for monitoring tool in the Central Asian countries.

  13. Monitoring the ecology of Bacillus during Daqu incubation, a fermentation starter, using culture-dependent and culture-independent methods.

    Science.gov (United States)

    Yan, Zheng; Zheng, Xiao-Wei; Han, Bei-Zhong; Han, Jian-Shu; Nout, M J Robert; Chen, Jing-Yu

    2013-05-01

    Daqu, a traditional fermentation starter, has been used to produce attractively flavored foods such as vinegar and Chinese liquor for thousands of years. Although Bacillus spp. are one of the dominant microorganisms in Daqu, more precise information is needed to reveal why and how Bacillus became dominant in Daqu, and next, to assess the impact of Bacillus sp. on Daqu and its derived products. We combined culture-dependent and culture-independent methods to study the ecology of Bacillus during Daqu incubation. Throughout the incubation, 67 presumptive Bacillus spp. isolates were obtained, 52 of which were confirmed by 16S rDNA sequencing. The identified organisms belonged to 8 Bacillus species: B. licheniformis, B. subtilis, B. amyloliquefaciens, B. cereus, B. circulans, B. megaterium, B. pumilus, and B. anthracis. A primer set specific for Bacillus and related genera was used in a selective PCR study, followed by a nested DGGE PCR targeting the V9 region of the 16S rDNA. Species identified from the PCR-DGGE fingerprints were related to B. licheniformis, B. subtilis, B. amyloliquefaciens, B. pumilus, B. benzoevorans, and B. foraminis. The predominant species was found to be B. licheniformis. Certain B. licheniformis strains exhibited potent antimicrobial activities. The greatest species diversity occurred at the Liangmei stage of Daqu incubation. To date, we lack sufficient knowledge of Bacillus distribution in Daqu. Elucidating the ecology of Bacillus during Daqu incubation would enable the impact of Bacillus on Daqu to be accessed, and the quality and stabilization of Daqu-derived products to be optimized.

  14. Fluidic system for long-term in vitro culturing and monitoring of organotypic brain slices

    DEFF Research Database (Denmark)

    Bakmand, Tanya; Troels-Smith, Ane R.; Dimaki, Maria

    2015-01-01

    Brain slice preparations cultured in vitro have long been used as a simplified model for studying brain development, electrophysiology, neurodegeneration and neuroprotection. In this paper an open fluidic system developed for improved long term culturing of organotypic brain slices is presented. ...

  15. Magnetic Suspension Technology Workshop

    International Nuclear Information System (INIS)

    Keckler, C.R.; Groom, N.J.; Britcher, C.P.

    1993-01-01

    In order to identify the state of magnetic suspension technology in such areas as rotating systems, pointing of experiments or subsystems, payload isolation, and superconducting materials, a workshop on Magnetic Suspension Technology was held at the Langley Research Center in Hampton, Virginia, on 2-4 Feb. 1988. The workshop included five technical sessions in which a total of 24 papers were presented. The technical sessions covered the areas of pointing, isolation, and measurement, rotating systems, modeling and control, and superconductors. A list of attendees is provided. Separate abstracts have been prepared for articles from this report

  16. Monitoring ground deformation of cultural heritage sites using UAVs and geodetic techniques: the case study of Choirokoitia, JPI PROTHEGO project

    Science.gov (United States)

    Themistocleous, Kyriacos; Danezis, Chris; Mendonidis, Evangelos; Lymperopoulou, Efstathia

    2017-10-01

    This paper presents the integrated methods using UAVs and geodetic techniques to monitor ground deformation within the Choirokoitia UNESCO World Heritage Site in Cyprus. The Neolithic settlement of Choirokoitia, occupied from the 7th to the 4th millennium B.C., is one of the most important prehistoric sites in the eastern Mediterranean. The study is conducted under the PROTHEGO (PROTection of European Cultural HEritage from GeO-hazards) project, which is a collaborative research project funded in the framework of the Joint Programming Initiative on Cultural Heritage and Global Change (JPICH) - Heritage Plus in 2015-2018 (www.prothego.eu) and through the Cyprus Research Promotion Foundation. PROTHEGO aims to make an innovative contribution towards the analysis of geo-hazards in areas of cultural heritage, and uses novel space technology based on radar interferometry to retrieve information on ground stability and motion in the 400+ UNESCO's World Heritage List monuments and sites of Europe. The field measurements collected at the Choirokoitia site will be later compared with SAR data to verify micro-movements in the area to monitor potential geo-hazards. The site is located on a steep hill, which makes it vulnerable to rock falls and landslides.

  17. Simultaneous and quantitative monitoring of co-cultured Pseudomonas aeruginosa and Staphylococcus aureus with antibiotics on a diffusometric platform

    Science.gov (United States)

    Chung, Chih-Yao; Wang, Jhih-Cheng; Chuang, Han-Sheng

    2017-04-01

    Successful treatments against bacterial infections depend on antimicrobial susceptibility testing (AST). However, conventional AST requires more than 24 h to obtain an outcome, thereby contributing to high patient mortality. An antibiotic therapy based on experiences is therefore necessary for saving lives and escalating the emergence of multidrug-resistant pathogens. Accordingly, a fast and effective drug screen is necessary for the appropriate administration of antibiotics. The mixed pathogenic nature of infectious diseases emphasizes the need to develop an assay system for polymicrobial infections. On this basis, we present a novel technique for simultaneous and quantitative monitoring of co-cultured microorganisms by coupling optical diffusometry with bead-based immunoassays. This simple integration simultaneously achieves a rapid AST analysis for two pathogens. Triple color particles were simultaneously recorded and subsequently analyzed by functionalizing different fluorescent color particles with dissimilar pathogen-specific antibodies. Results suggested that the effect of the antibiotic, gentamicin, on co-cultured Pseudomonas aeruginosa and Staphylococcus aureus was effectively distinguished by the proposed technique. This study revealed a multiplexed and time-saving (within 2 h) platform with a small sample volume (~0.5 μL) and a low initial bacterial count (50 CFU per droplet, ~105 CFU/mL) for continuously monitoring the growth of co-cultured microorganisms. This technique provides insights into timely therapies against polymicrobial diseases in the near future.

  18. Crewbot Suspension Design

    Science.gov (United States)

    Wood, Nathan A.

    2005-01-01

    Planetary Surface Robot Work Crews (RWC) represent a new class of construction robots for future deployment in planetary exploration. Rovers currently being used for the RWC platform lack the load carrying capabilities required in regular work. Two new rovers, dubbed CrewBots, being designed in JPL's Planetary Robotics Lab specifically for RWC applications greatly increase the load carrying capabilities of the platform. A major component of the rover design was the design of the rocker type suspension, which increases rover mobility. The design of the suspension for the Crewbots departed from the design of recent rovers. While many previous rovers have used internal bevel gear differentials, the increased load requirements of the Crewbots calls for a more robust system. The solution presented is the use of an external modified three-bar, slider-linkage, rocker-style suspension that increases the moment arm of the differential. The final product is a suspension system capable of supporting the extreme loading cases the RWC platform presents, without consuming a large portion of the Crewbots' internal space.

  19. Observation of antioxidant activity of leaves, callus and suspension ...

    African Journals Online (AJOL)

    GREGORY

    2011-12-16

    Dec 16, 2011 ... Antioxidant activity and phenolic compound was found in Justicia gendarussa via total phenolic content (TPC) and α,α-diphenyl-β-pycrilhydrazil hydrate (DPPH) radical scavenging assays. The assays were applied on aqueous and methanolic extracts of leaves, callus culture and cell suspension culture.

  20. Enhancement of 20-hydroxyecdysone production in cell suspension ...

    African Journals Online (AJOL)

    ... most effective. The maximum 20-hydroxyecdysone productivity of about 1.31 mg/L/day was observed in culture with 10 mg/L 7-dehydrocholesterol. This data is the first indication that 7-dehydrocholesterol and ergosterol feeding are effective precursors for 20-hydroxyecdysone formation in plant cell suspension culture.

  1. Observation of antioxidant activity of leaves, callus and suspension ...

    African Journals Online (AJOL)

    Antioxidant activity and phenolic compound was found in Justicia gendarussa via total phenolic content (TPC) and ,-diphenyl--pycrilhydrazil hydrate (DPPH) radical scavenging assays. The assays were applied on aqueous and methanolic extracts of leaves, callus culture and cell suspension culture of J. gendarussa. Callus ...

  2. Mevastatin-induced inhibition of cell growth in avocado suspension ...

    African Journals Online (AJOL)

    Cell suspension cultures were established using soft, friable callus derived from nucellar tissue of 'Hass' avocado (Persea americana Mill.) seed from fruit harvested 190 days after full bloom. Cell cultures were maintained in liquid medium supplemented with naphthalene acetic acid (NAA), isopentenyl adenine (iP) and ...

  3. Design of a microscopic electrical impedance tomography system for 3D continuous non-destructive monitoring of tissue culture.

    Science.gov (United States)

    Lee, Eun Jung; Wi, Hun; McEwan, Alistair Lee; Farooq, Adnan; Sohal, Harsh; Woo, Eung Je; Seo, Jin Keun; Oh, Tong In

    2014-10-06

    Non-destructive continuous monitoring of regenerative tissue is required throughout the entire period of in vitro tissue culture. Microscopic electrical impedance tomography (micro-EIT) has the potential to monitor the physiological state of tissues by forming three-dimensional images of impedance changes in a non-destructive and label-free manner. We developed a new micro-EIT system and report on simulation and experimental results of its macroscopic model. We propose a new micro-EIT system design using a cuboid sample container with separate current-driving and voltage sensing electrodes. The top is open for sample manipulations. We used nine gold-coated solid electrodes on each of two opposing sides of the container to produce multiple linearly independent internal current density distributions. The 360 voltage sensing electrodes were placed on the other sides and base to measure induced voltages. Instead of using an inverse solver with the least squares method, we used a projected image reconstruction algorithm based on a logarithm formulation to produce projected images. We intended to improve the quality and spatial resolution of the images by increasing the number of voltage measurements subject to a few injected current patterns. We evaluated the performance of the micro-EIT system with a macroscopic physical phantom. The signal-to-noise ratio of the developed micro-EIT system was 66 dB. Crosstalk was in the range of -110.8 to -90.04 dB. Three-dimensional images with consistent quality were reconstructed from physical phantom data over the entire domain. From numerical and experimental results, we estimate that at least 20 × 40 electrodes with 120 μm spacing are required to monitor the complex shape of ingrowth neotissue inside a scaffold with 300 μm pore. The experimental results showed that the new micro-EIT system with a reduced set of injection current patterns and a large number of voltage sensing electrodes can be potentially used for tissue

  4. Car suspension system monitoring under road conditions

    Science.gov (United States)

    Fedotov, A. I.; Kuznetsov, N. Y.; Lysenko, A. V.; Vlasov, V. G.

    2017-12-01

    The paper describes an advanced gyro-based measuring system comprising a CGV-4K central vertical gyro and a G-3M gyrocompass. The advanced system provides additional functions that help measure unsprung mass rotation angles about a vertical axis, rolling angles, trim angles and movements of the unsprung masses of the front (ap and al) and rear b axes when a car wheel hits a single obstruction. The paper also describes the operation of the system, which measures movements of unsprung masses about the body of a car when it hits a single obstruction. The paper presents the dependency diagrams ap = f(t) and al = f(t) for front and rear wheels respectively, as well as b = f(t) for a rear left wheel, which were determined experimentally. Test results for a car equipped with an advanced gyro-based measuring system moving around a circle can form a basis for developing a mathematical model of the process.

  5. Comparison of spectroscopy technologies for improved monitoring of cell culture processes in miniature bioreactors

    DEFF Research Database (Denmark)

    Rowland-Jones, Ruth C.; van der Berg, Franciscus Winfried J; Racher, Andrew J.

    2017-01-01

    Cell culture process development requires the screening of large numbers of cell lines and process conditions. The development of miniature bioreactor systems has increased the throughput of such studies; however, there are limitations with their use. One important constraint is the limited numbe...

  6. Cultural

    Science.gov (United States)

    Wilbur F. LaPage

    1971-01-01

    A critical look at outdoor recreation research and some underlying premises. The author focuses on the concept of culture as communication and how it influences our perception of problems and our search for solutions. Both outdoor recreation and science are viewed as subcultures that have their own bodies of mythology, making recreation problems more difficult to...

  7. Monitoring Cultural Heritage Sites with Advanced Multi-Temporal InSAR Technique: The Case Study of the Summer Palace

    Directory of Open Access Journals (Sweden)

    Panpan Tang

    2016-05-01

    Full Text Available Cultural heritage sites are rare and irreplaceable wealth of human civilization. The majority of them are becoming unstable due to a combination of human and natural disturbances. High-precision, efficient deformation monitoring facilitates the early recognition of potential risks and enables preventive diagnosis of heritage sites. In this study, an advanced Multi-Temporal Interferometric Synthetic Aperture Radar (MTInSAR approach was developed by jointly analyzing Persistent Scatterers (PSs and Distributed Scatterers (DSs using high-resolution SAR images. Taking the World Heritage Site of Summer Palace in Beijing as the experimental site, deformation resulting from PSs/DSs showed that overall the site was generally stable except for specific areas and/or monuments. Urbanization (construction and demolition triggered new subsidence in the vicinity of East and South Gate of the site. Slight to moderate (mm/cm-level instabilities of ruins and monuments on Longevity Hill were detected, perhaps due to a combination of destructive anthropogenic activities and long-term natural decay. Subsidence was also detected along the Kunming Lakeside and was probably attributable to variation of the groundwater level, excessive visitor numbers as well as lack of maintenance. This study presents the potential of the MTInSAR approach for the monitoring and conservation of cultural heritage sites.

  8. CONCEPTUAL PRECONDITIONS OF SANOGENETICAL MONITORING OF PERSONS ENGAGED IN PHYSICAL CULTURE AND SPORTS

    OpenAIRE

    Romanchuk, A.P.

    2015-01-01

    The modern representations about human biology, theoretical medicine and psychology are reduced to that the individualized approach based on the account of typological features is necessary for optimization of the condition of a person. The purpose of occupations by sports is the development of physical qualities of the person, perfection of technical-tactical skills for achievement of the maximal sports result. At the same time the purpose of occupations by physical culture is the mainten...

  9. Monitoring the sensitivity of the oligochaete Tubifex tubifex in laboratory cultures using three toxicants.

    Science.gov (United States)

    Maestre, Zuriñe; Martinez-Madrid, Maite; Rodriguez, Pilar

    2009-11-01

    Acute toxicity tests were conducted to assess the sensitivity of laboratory cultured Tubifex tubifex (Annelida, Clitellata), as an intralaboratory quality assurance requirement for the test organisms used in sediment chronic bioassays. Worms were exposed to cadmium, copper, and chromium in water-only 96-h tests, in single metal exposure, over a three-year period. The lethal concentration for the x% of the exposed population (LC(x)), the No-Observable-Effect Concentration (NOEC) and the Lowest-Observable-Effect Concentration (LOEC) were estimated. Based on LC(50) values, the rank of toxicity was Cu(+2)>Cd(+2)>Cr(+6). Sensitivity remained relatively constant during the research period and was comparable with the sensitivity of the same culture stock examined more than 10 years ago. This observed consistency in sensitivity indicates that comparisons between different sediment chronic bioassays conducted during the study period, using T. tubifex worms from the same culture stock, was appropriate. It is necessary to control both worm biomass and age for the acute test in order to standardize the test-organism conditions in intralaboratory quality assurance programs.

  10. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz

    2006-01-01

    Full Text Available Es un estudio cualitativo que adoptó como referencial teorico-motodológico la antropología y la etnografía. Presenta las experiencias vivenciadas por mujeres de una comunidad en el proceso salud-enfermedad, con el objetivo de comprender los determinantes sócio-culturales e históricos de las prácticas de prevención y tratamiento adoptados por el grupo cultural por medio de la entrevista semi-estructurada. Los temas que emergieron fueron: la relación entre la alimentación y lo proceso salud-enfermedad, las relaciones con el sistema de salud oficial y el proceso salud-enfermedad y lo sobrenatural. Los dados revelaron que los moradores de la comunidad investigada tienen un modo particular de explicar sus procedimientos terapéuticos. Consideramos que es papel de los profesionales de la salud en sus prácticas, la adopción de abordajes o enfoques que consideren al individuo en su dimensión sócio-cultural e histórica, considerando la enorme diversidad cultural en nuestro país.

  11. Particle interactions in concentrated suspensions

    International Nuclear Information System (INIS)

    Mondy, L.A.; Graham, A.L.; Abbott, J.R.; Brenner, H.

    1993-01-01

    An overview is presented of research that focuses on slow flows of suspensions in which colloidal and inertial effects are negligibly small. The authors describe nuclear magnetic resonance imaging experiments to quantitatively measure particle migration occurring in concentrated suspensions undergoing a flow with a nonuniform shear rate. These experiments address the issue of how the flow field affects the microstructure of suspensions. In order to understand the local viscosity in a suspension with such a flow-induced, spatially varying concentration, one must know how the viscosity of a homogeneous suspension depends on such variables as solids concentration and particle orientation. The authors suggest the technique of falling ball viscometry, using small balls, as a method to determine the effective viscosity of a suspension without affecting the original microstructure significantly. They also describe data from experiments in which the detailed fluctuations of a falling ball's velocity indicate the noncontinuum nature of the suspension and may lead to more insights into the effects of suspension microstructure on macroscopic properties. Finally, they briefly describe other experiments that can be performed in quiescent suspensions (in contrast to the use of conventional shear rotational viscometers) in order to learn more about boundary effects in concentrated suspensions

  12. Feasibility of Focused Beam Reflectance Measurement (FBRM) for Analysis of Pharmaceutical Suspensions in Preclinical Development.

    Science.gov (United States)

    Dave, Kaushalkumar; Luner, Paul E; Forness, Cecile; Baker, Doris; Jankovsky, Corinne; Chen, Shirlynn

    2018-01-01

    This study examined the use of focused beam reflectance measurement (FBRM) for qualitative and quantitative analysis of pharmaceutical suspensions with particular application to toxicology supply preparations for use in preclinical studies. Aqueous suspensions of ibuprofen were used as prototype formulations. Initial experiments were conducted to examine the effects of operational conditions including FBRM probe angle, probe location, and mixing (method and rate of mixing) on the FBRM analysis. Once experimental conditions were optimized, the homogeneity and sedimentation-redispersion of particles in the suspensions were assessed. Ibuprofen suspension under continuous agitation was monitored using FBRM for 60 h to study particle size change over time. Another study was performed to determine if particle count rates obtained by FBRM could be correlated to suspension concentration. The location and the angle of the FBRM probe relative to the beaker contents, and the rate and the method of mixing the suspension were found to be sensitive parameters during FBRM analysis. FBRM was able to monitor the process of particle sedimentation in the suspension. The attrition of ibuprofen particles was detectable by FBRM during prolonged stirring with an increase in the number of smaller particles and decrease in the number of larger particles. A strong correlation was observed between particle count rate by FBRM and ibuprofen concentration in the suspension. Also, change in content uniformity in the suspension at different locations of the beaker was represented by FBRM particle count. Overall, FBRM has potential to be a useful tool for qualitative and quantitative analysis of pharmaceutical suspensions.

  13. Selection, application and monitoring of Lactobacillus paracasei strains as adjunct cultures in the production of Gouda-type cheeses.

    Science.gov (United States)

    Van Hoorde, Koenraad; Van Leuven, Isabelle; Dirinck, Patrick; Heyndrickx, Marc; Coudijzer, Kathleen; Vandamme, Peter; Huys, Geert

    2010-12-15

    Raw milk cheeses have more intense flavours than cheeses made from pasteurized milk and harbour strains with potential adjunct properties. Two Lactobacillus paracasei strains, R-40926 and R-40937, were selected as potential adjunct cultures from a total of 734 isolates from good quality artisan raw milk Gouda-type cheeses on the basis of their prevalence in different cheese types and/or over several production batches, safety and technological parameters. Conventional culturing, isolation and identification and a combined PCR-DGGE approach using total cheese DNA extracts and DNA extracts obtained from culturable fractions were employed to monitor viability of the introduced adjuncts and their effect on the cheese microbiota. The control cheese made without adjuncts was dominated by members of the starter, i.e. Lactococcus lactis and Leuconostoc pseudomesenteroides. In the cheeses containing either R-40926 or R-40937, the respective adjuncts increased in number as ripening progressed indicating that both strains are well adapted to the cheese environment and can survive in a competitive environment in the presence of a commercial starter culture. Principal component analysis of cheese volatiles determined by steam distillation-extraction and gas chromatography-mass spectrometry could differentiate cheeses made with different concentrations of adjunct R-40926 from the control cheese, and these differences could be correlated to the proteolytic and lipolytic properties of this strain. Collectively, results from microbiological and metabolic analyses indicate that the screening procedure followed throughout this study was successful in delivering potential adjunct candidates to enrich or extend the flavour palette of artisan Gouda-type cheeses under more controlled conditions. Copyright © 2010 Elsevier B.V. All rights reserved.

  14. Monitoring of water in soil in asparagus irrigated culture in Vale do Sao Francisco, Pernambuco, Brazil

    International Nuclear Information System (INIS)

    Antonino, Antonio C. Dantas; Sampaio, Everardo V.S.B.; Dall' Ollio, Attilio; Bernardo, Ana L. Alves; Audry, Pierre

    1996-08-01

    For many years the brazilian government has inactivated the implantation of irrigated areas in the Sao Francisco valley, obtaining high productivity.After the most appropriated areas having been occupied, the irrigation of second choice soils, usually more shallower and more clay is been tried. In one of these areas, the productivity of asparagus is less than the expected. trying to improve productivity by optimization of irrigation, the movement of water on soil and plants is being monitored far the last year and a half. the main results are shown with emphasis on the raining season, the most problematic

  15. Monitoring of Monthly Price Changes of Cultured Fishes in Sinop Province Between 2012 and 2013

    Directory of Open Access Journals (Sweden)

    Birol Baki

    2017-09-01

    Full Text Available The aim of this study was to determine the monthly price changes of cultured fish which were offered for sale in retail fish outlets in Sinop, Turkey. The fish enterprises were determined by stratified sampling method. The enterprises were grouped as I (the most, II (moderate and III (the least according to the amount of their sales. Each group was represented by two different fish enterprises. During the study, prices of the same fish species were obtained twice a month at the same time of the day and the data were calculated as average ± SD. The differences between the groups were determined by one-way variance analysis (ANOVA and the price relation level between species was evaluated by the correlation analysis using IBM SPSS 21 Statisticssoftware. It was determined that sea bream (Sparus aurata, sea bass (Dicentrarhus labrax, rainbow trout (Oncorhyncus mykiss (portion size and kilos and salmon (Salmo salar (import were offered for sale in fish markets as cultured fish species. These species were sold during the fishing ban period (April-August. They were also sold starting from September when the fishing ban period ended up and the fishery products were at their lowest. For all the species, the differences between the pricesice were not significant between the sampled enterprises. The average values of the current prices were higher than that of 2008-2013 period. A negative strong relation was determined between the rainbow trout and the salmon.

  16. Miniaturized Biosensors to Preserve and Monitor Cultural Heritage: from Medical Diagnosis to Conservation Diagnosis.

    Science.gov (United States)

    Sciutto, Giorgia; Zangheri, Martina; Anfossi, Laura; Guardigli, Massimo; Prati, Silvia; Mirasoli, Mara; Di Nardo, Fabio; Baggiani, Claudio; Mazzeo, Rocco; Roda, Aldo

    2018-04-10

    We have exploited the point-of-care testing concept to design and develop portable and cheap bioanalytical systems that can be used on-site by conservators. These systems employ lateral flow immunoassays to simultaneously detect two proteins (ovalbumin and collagen) in artworks. For an in-depth study on the application of these portable biosensors, both chemiluminescent and colorimetric detections were developed and compared in terms of sensitivity and feasibility. The chemiluminescent system displayed the best analytical performance (i.e., two orders of magnitude lower limits of detection than the colorimetric system). To simplify its use a disposable cartridge was designed ad hoc for this specific application. These results highlight the enormous potential of these inexpensive, easy-to-use, and minimally invasive diagnostic tools for conservators in the cultural heritage field. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. APPLICATION OF THE QUALITY NORMS TO THE MONITORING AND THE PREVENTIVE CONSERVATION ANALYSIS OF THE CULTURAL HERITAGE

    Directory of Open Access Journals (Sweden)

    Massimo Andretta

    2015-06-01

    Full Text Available In recent years, the study of the indoor microclimate has assumed increasing importance, especially for the problems associated with the conservation of the cultural heritage housed in museums, galleries and libraries. In this paper, we describe the most important national standards relative to the procedures for the measurements and the analysis of the environmental conditions regarding the preservation of the works of art. These methods are related to the measurement techniques, which have to be applied for monitoring and analyzing the microclimatic conditions of museums, galleries and archives; these norms report, also, the threshold reference values for optimal climatic conditions. Furthermore, we present some considerations on the importance and on the foundations of the proposed scientific/methodological approaches. Finally, we have done a reasoned analysis on some reference values reported by the international regulations with some considerations on the possible chemical/physical mechanisms of degradation of the valuable objects.

  18. Development of a technique based on multi-spectral imaging for monitoring the conservation of cultural heritage objects.

    Science.gov (United States)

    Marengo, Emilio; Manfredi, Marcello; Zerbinati, Orfeo; Robotti, Elisa; Mazzucco, Eleonora; Gosetti, Fabio; Bearman, Greg; France, Fenella; Shor, Pnina

    2011-11-14

    A new approach for monitoring the state of conservation of cultural heritage objects surfaces is being developed. The technique utilizes multi-spectral imaging, multivariate analysis and statistical process control theory for the automatic detection of a possible deterioration process, its localization and identification, and the wavelengths most sensitive to detecting this before the human eye can detect the damage or potential degradation changes occur. A series of virtual degradation analyses were performed on images of parchment in order to test the proposed algorithm in controlled conditions. The spectral image of a Dead Sea Scroll (DSS) parchment, IAA (Israel Antiquities Authority) inventory plate # 279, 4Q501 Apocryphal Lamentations B, taken during the 2008 Pilot of the DSS Digitization Project, was chosen for the simulation. Copyright © 2011 Elsevier B.V. All rights reserved.

  19. Development of a multiplexer for an automatic data acquisition system for the control and monitoring of microbiological cultures

    Energy Technology Data Exchange (ETDEWEB)

    Morales Rondon, A.; Paredes Puente, J.; Arana Alonso, S.

    2016-07-01

    An automatic data acquisition system has been developed for the control and monitoring of microbiological cultures. Turning an otherwise time-consuming process into a smooth one, by allowing the researcher to set the parameters at the beginning of the experiment and move on into the next task. The development of the hardware and software are key to achieving this system. The mux is custom-made with 22 channels, light weight therefore easy to move around the lab. Furthermore, the software allows the researcher to check the measurements in real-time. It is based on virtual instrumentation software therefore new features can be added easily, thus, the mux is capable of adapting to the scientist necessities. (Author)

  20. Reproducibility of measurement of the environmental carbon-14 samples prepared by the gel suspension method

    International Nuclear Information System (INIS)

    Ohura, Hirotaka; Wakabayashi, Genichiro; Nakamura, Kouji; Okai, Tomio; Matoba, Masaru; Kakiuchi, Hideki; Momoshima, Noriyuki; Kawamura, Hidehisa.

    1997-01-01

    Simple liquid scintillation counting technique for the assay of 14 C in the environment was developed. This technique was done by using gel suspension method, in which sample preparation is very simple and requires no special equipments. The reproducibility of this technique was considered and it was shown that the gel suspension method had enough reproducibility to monitor the environmental 14 C. (author)

  1. Importância do co-cultivo com fibroblastos de camundongo 3T3 para estabelecer cultura de suspensão de células epiteliais do limbo humano Importance of 3T3 feeder layer to establish epithelial cultures from cell suspension obtained from corneo-scleral rims

    Directory of Open Access Journals (Sweden)

    Priscila Cardoso Cristovam

    2008-10-01

    Full Text Available OBJETIVO: Avaliar a importância da presença de células 3T3 para estabelecer cultura de suspensão de células epiteliais do limbo obtido de rimas córneo-esclerais. MÉTODOS: Rimas de diferentes doadores tiveram seus estroma posterior e endotélio removidos (n=6. Cada rima foi dividida em três segmentos iguais, que foram colocados em cultura em três diferentes condições: um segmento foi colocado na placa de cultura com o lado epitelial para cima (Grupo A. Os dois segmentos restantes foram tripsinizados e a suspensão de células obtida foi cultivada com (Grupo B ou sem (Grupo C células 3T3 irradiadas. As células foram mantidas em meio de cultura "supplemental hormonal epithelial médium" (SHEM, a migração epitelial e a formação de clones nos grupos A, B e C foram avaliadas pela microscopia de contraste de fase e por coloração pela rodamina B. Os resultados foram comparados estatisticamente. RESULTADOS: O crescimento de células epiteliais foi observado em 4/6 rimas (Grupo A. Todas as suspensões de células epiteliais que foram cultivadas com células 3T3 (Grupo B formaram clones. Nenhuma adesão ou formação de clones verdadeiros (holo ou meroclones foi observada na cultura de células que foi cultivada sem 3T3 (Grupo C (p=0,009. CONCLUSÕES: Suspensão de células epiteliais límbicas obtidas de rimas córneo-esclerais no modelo utilizado precisa ser cultivada com células 3T3 para formar clones e estabelecer colônias epiteliais com perspectivas para uso terapêutico na reconstrução da superfície ocular.PURPOSE: To evaluate the importance of the presence of 3T3 fibroblasts for establishing limbal epithelial cultures from cell suspension obtained from corneo-scleral rims (CSR. METHODS: Corneo-scleral rims from different donors (n=6 had their posterior stroma and endothelium stripped away. Each corneo-scleral rim was divided into three equal segments that were set up in tissue culture in three different conditions: one of the

  2. Monitoring utilizations of amino acids and vitamins in culture media and Chinese hamster ovary cells by liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Qiu, Jinshu; Chan, Pik Kay; Bondarenko, Pavel V

    2016-01-05

    Monitoring amino acids and vitamins is important for understanding human health, food nutrition and the culture of mammalian cells used to produce therapeutic proteins in biotechnology. A method including ion pairing reversed-phase liquid chromatography with tandem mass spectrometry was developed and optimized to quantify 21 amino acids and 9 water-soluble vitamins in Chinese hamster ovary (CHO) cells and culture media. By optimizing the chromatographic separation, scan time, monitoring time window, and sample preparation procedure, and using isotopically labeled (13)C, (15)N and (2)H internal standards, low limits of quantitation (≤0.054 mg/L), good precision (vitamins accumulated continuously during the culture period, suggesting that they were fed in access. The method serves as an effective tool for the development and optimization of mammalian cell cultures. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. THE WATER QUALITY MONITORING IN THE RECIRCULATING SYSTEMS FOR INTENSIVE STURGEON CULTURE

    Directory of Open Access Journals (Sweden)

    C. SAVIN

    2009-10-01

    Full Text Available The major modification of those parameters go to apparition of the biological and physico-chemical imbalance of water environment who produce disturbance to normal way of vital function, that respond through the primary physiological modification (endocrine follow by secondary physiological modification (hormonal, with direct influence over the growing parameters and the pathological condition. The rearing experiments were lasting 60 days and have been achieved on young beluga sturgeon (Huso huso in two different locations, with two different recirculating pilot systems. Was analyzed the main physico-chemical parameters of sturgeon culture, obtaining 23 - 25°C temperature, 4 – 5 mg/l dissolved oxygen, 7,6 upH pH, 0,02 – 0,23 mg/l ammonia, 0,13 – 0,17 mg/l ammonium, 0,09 – 0,45 mg/l nitrite and 10,55 – 72,06 mg/l nitrate.

  4. Flow properties of concentrated suspensions

    Science.gov (United States)

    Hattori, K.; Izumi, K.

    1984-04-01

    The viscosity and flow behavior of a concentrated suspension, with special emphasis on fresh concrete containing a superplasticizer, is analyzed according to Newton's law of viscosity. The authors interpreted Newton's law in a new way, and explain non-Newton flow from Newton's law. The outline of this new theory is given. Viscosity of suspensions, and the effect of dispersants are analyzed.

  5. Biodiversity within hot spring microbial mat communities: molecular monitoring of enrichment cultures

    Science.gov (United States)

    Ward, D. M.; Santegoeds, C. M.; Nold, S. C.; Ramsing, N. B.; Ferris, M. J.; Bateson, M. M.

    1997-01-01

    We have begun to examine the basis for incongruence between hot spring microbial mat populations detected by cultivation or by 16S rRNA methods. We used denaturing gradient gel electrophoresis (DGGE) to monitor enrichments and isolates plated therefrom. At near extincting inoculum dilutions we observed Chloroflexus-like and cyanobacterial populations whose 16S rRNA sequences have been detected in the 'New Pit' Spring Chloroflexus mat and the Octopus Spring cyanobacterial mat. Cyanobacterial populations enriched from 44 to 54 degrees C and 56 to 63 degrees C samples at near habitat temperatures were similar to those previously detected in mat samples of comparable temperatures. However, a lower temperature enrichment from the higher temperature sample selected for the populations found in the lower temperature sample. Three Thermus populations detected by both DGGE and isolation exemplify even more how enrichment may bias our view of community structure. The most abundant population was adapted to the habitat temperature (50 degrees C), while populations adapted to 65 degrees C and 70 degrees C were 10(2)- and 10(4)-fold less abundant, respectively. However, enrichment at 70 degrees C favored the least abundant strain. Inoculum dilution and incubation at the habitat temperature favored the more numerically relevant populations. We enriched many other aerobic chemoorganotrophic populations at various inoculum dilutions and substrate concentrations, most of whose 16S rRNA sequences have not been detected in mats. A common feature of numerically relevant cyanobacterial, Chloroflexus-like and aerobic chemorganotrophic populations, is that they grow poorly and resist cultivation on solidified medium, suggesting plating bias, and that the medium composition and incubation conditions may not reflect the natural microenvironments these populations inhabit.

  6. Sensor technologies and non-destructive monitoring for dampness diagnosis in cultural heritage

    Science.gov (United States)

    Inmaculada Martínez Garrido, María; Gómez Heras, Miguel; Fort González, Rafael; Valles Iriso, Javier; José Varas Muriel, María

    2016-04-01

    This work presents a case study based on results of monitoring campaigns developed in San Juan Bautista church in Talamanca de Jarama (Madrid, Spain). This Church was built in the twelfth-thirteenth centuries (Romanesque style) with dolostone ashlars. It was reconstructed in the sixteenth century (Renaissance style) with rubble stone and mortar, brick and an earth fill. Different sections on walls and floors (north and south oriented) have been selected based on a preliminary study of moisture distribution on stone and masonry wall. The behavior of different materials has been studied according to the influence of indoor (microclimatic conditions) and outdoor conditions (weather conditions) and taking into account constructive facts. Several sensing technologies as dataloggers and wireless sensor networks (WSN) together to other non invasive techniques as thermal imaging, portable moisture meter, electrical resistivity tomography (ERT) and ground-penetrating radar (GPR) have been conducted. By means of this study it has been possible to establish an analysis methodology to determine the dampness origin in each case. Conclusions related to the each technique according to its effectiveness in the detection of decay problems have been established. Research funded by Geomateriales 2(S2013/MIT-2914) and Deterioration of stone materials in the interior of historic buildings as a result induced variation of its microclimate (CGL2011-27902) projects. The cooperation received from the Complutense University of Madrid's Research Group Alteración y Conservación de los Materiales Pétreos del Patrimonio (ref. 921349), the Laboratory Network in Science and Technology for Heritage Conservation (RedLabPat, CEI Moncloa) and the Diocese of Alcalá is gratefully acknowledged.

  7. Recording, Visualization and Documentation of 3D Spatial Data for Monitoring Topography in Areas of Cultural Heritage

    Science.gov (United States)

    Maravelakis, Emmanouel; Konstantaras, Antonios; Axaridou, Anastasia; Chrysakis, Ioannis; Xinogalos, Michalis

    2014-05-01

    . allowing them to interchange their knowledge, findings and observations at different time frames. Results outline the successful application of the above systems in certain Greek areas of important cultural heritage [3,11] were significant efforts are being made for their preservation through time. Acknowledgement The authors wish to thank the General Secretariat for Research and Technology of Ministry of Education and Religious Affairs, Culture and Sports in Greece for their financial support via program Cooperation: Partnership of Production and Research Institutions in Small and Medium Scale Projects, Project Title: "3D-SYSTEK - Development of a novel system for 3D Documentation, Promotion and Exploitation of Cultural Heritage Monuments via 3D data acquisition, 3D modeling and metadata recording". Keywords spatial data, land degradation monitoring, 3D modeling and visualization, terrestrial laser scanning, documentation and metadata repository, protection of cultural heritage References [1] Shalaby, A., and Tateishi, R.: Remote sensing and GIS for mapping and monitoring land cover and land-use changes in the northwestern coastal zone of egypt. Applied Geography, 27(1), 28-41, (2007) [2] Poesen, J. W. A., and Hooke, J. M.: Erosion, flooding and channel management in mediterranean environments of southern europe. Progress in Physical Geography, 21(2), 157-199, (1997) [3] Maravelakis, E., Bilalis, N., Mantzorou, I., Konstantaras, A., Antoniadis, A.: 3D modeling of the oldest olive tree of the world. IJCER 2(2), 340-347 (2012) [4] Manferdini, A.M., Remondino, F.: Reality-Based 3D Modeling, Segmentation and Web- Based Visualization. In: Ioannides, M., Fellner, D., Georgopoulos, A., Hadjimitsis, D.G. (eds.) EuroMed 2010. LNCS, vol. 6436, pp. 110-124. Springer, Heidelberg (2010) [5] Tapete, D., Casagli, N., Luzi, G., Fanti, R., Gigli, G., Leva, D.: Integrating radar and laserbased remote sensing techniques for monitoring structural deformation of archaeological monuments

  8. Mapping and characterisation of the sorghum cell suspension ...

    African Journals Online (AJOL)

    Here we reported the first secretomic study of sorghum (Sorghum bicolor), a naturally drought tolerant cereal crop. In this study, we used a gel-based proteomic approach in combination with mass spectrometry to separate and identify proteins secreted into the culture medium of sorghum cell suspensions, a first step ...

  9. Monitoring sputum culture in resected esophageal cancer patients with preoperative treatment.

    Science.gov (United States)

    Kosumi, K; Baba, Y; Yamashita, K; Ishimoto, T; Nakamura, K; Ohuchi, M; Kiyozumi, Y; Izumi, D; Tokunaga, R; Harada, K; Shigaki, H; Kurashige, J; Iwatsuki, M; Sakamoto, Y; Yoshida, N; Watanabe, M; Baba, H

    2017-12-01

    Pneumonia is a major cause of postesophagectomy mortality and worsens the long-term survival in resected esophageal cancer patients. Moreover, preoperative treatments such as chemotherapy or chemoradiotherapy (which have recently been applied worldwide) might affect the bacterial flora of the sputum. To investigate the association among preoperative treatments, the bacterial flora of sputum, and the clinical and pathological features in resected esophageal cancer patients, this study newly investigates the effect of preoperative treatments on the bacterial flora of sputum. We investigated the association among preoperative treatments, the bacterial flora of sputum, and clinical and pathological features in 163 resected esophageal cancer patients within a single institution. Pathogenic bacteria such as Candida (14.1%), Staphylococcus aureus (6.7%), Enterobacter cloacae (6.1%), Haemophilus parainfluenzae (4.9%), Klebisiella pneumoniae (3.7%), Methicillin-resistant Staphylococcus aureus (MRSA) (3.7%), Pseudomonas aeruginosa (2.5%), Escherichia coli (1.8%), Streptococcus pneumoniae (1.8%), and Haemophilus influenzae (1.2%) were found in the sputum. The pathogen detection rate in the present study was 34.3% (56/163). In patients with preoperative chemotherapy and chemoradiotherapy, the indigenous Neisseria and Streptococcus species were significantly decreased (P= 0.04 and P= 0.04). However, the detection rates of pathogenic bacteria were not associated with preoperative treatments (all P> 0.07). There was not a significant difference of hospital stay between the sputum-monitored patients and unmonitored patients (35.5 vs. 49.9 days; P= 0.08). Patients undergoing preoperative treatments exhibited a significant decrease of indigenous bacteria, indicating that the treatment altered the bacterial flora of their sputum. This finding needs to be confirmed in large-scale independent studies or well-designed multicenter studies. © The Authors 2017. Published by Oxford

  10. Strategies to Suspension Serum-Free Adaptation of Mammalian Cell Lines for Recombinant Glycoprotein Production.

    Science.gov (United States)

    Caron, Angelo Luis; Biaggio, Rafael Tagé; Swiech, Kamilla

    2018-01-01

    Serum-free suspension cultures are preferably required for recombinant protein production due to its readiness in upstream/downstream processing and scale-up, therefore increasing process productivity and competitiveness. This type of culture replaces traditional cell culturing as the presence of animal-derived components may introduce lot-a-lot variability and adventitious pathogens to the process. However, adapting cells to serum-free conditions is challenging, time-consuming, and cell line and medium dependent. In this chapter, we present different approaches that can be used to adapt mammalian cell lines from an anchorage-dependent serum supplemented culture to a suspension serum-free culture.

  11. Modeling and monitoring of the acclimatization of conventional activated sludge to a biohydrogen producing culture by biokinetic control

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez-Morales, F.J.; Villasenor, J.; Infantes, D. [University of Castilla-La Mancha, ITQUIMA, Chemical Engineering Department, Avenida Camilo Jose Cela S/N. 13071 Ciudad Real (Spain)

    2010-10-15

    The purpose of this work was to study the acclimatization of activated sludge as it evolved into an acidogenic culture capable of producing volatile fatty acids and hydrogen from glucose. The experiments were conducted in a Sequential Batch Reactor with a pH of 5, a temperature of 35 C and in the absence of oxygen. Biomass, substrate and product concentrations were monitored during the experiment. The presence of aerobic, facultative and acidogenic microorganisms was determined for the initial seed, and their main kinetic and stoichiometric properties were determined. For the acidogenic microorganisms, {mu}{sub max}, b and Y{sub x} increased during the acclimatization process, reaching final values of 0.125 h{sup -1}, 0.025 h{sup -1}, 0.025 g mmol{sup -1}, respectively. However, for the facultative microorganisms, {mu}{sub max} and b decreased, whereas Y{sub x} remained constant during the acclimatization process, reaching final values of 0.032 h{sup -1}, 0.007 h{sup -1}, 0.017 g mmol{sup -1}, respectively. The evolution of the facultative organisms was significantly related to lactic acid production, whereas the evolution of the acidogenic microorganisms was related to hydrogen, acetic acid and butyric acid production. The biomass concentration of each group was estimated by modeling the experimental data. (author)

  12. 49 CFR 238.427 - Suspension system.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Suspension system. 238.427 Section 238.427... Equipment § 238.427 Suspension system. (a) General requirements. (1) Suspension systems shall be designed to... equipment. (2) Passenger equipment shall meet the safety performance standards for suspension systems...

  13. "Point de suspension"

    CERN Document Server

    2004-01-01

    CERN - Globe of Science and Innovation 20 and 21 October Acrobatics, mime, a cappella singing, projections of images, a magical setting... a host of different tools of a grandeur matching that of the Universe they relate. A camera makes a massive zoom out to reveal the multiple dimensions of Nature. Freeze the frame: half way between the infinitesimally small and the infinitesimally large, a man suspends his everyday life (hence the title "Point de Suspension", which refers to the three dots at the end of an uncompleted sentence) to take a glimpse of the place he occupies in the great history of the Universe. An unusual perspective on what it means to be a human being... This wondrous show in the Globe of Science and Innovation, specially created by the Miméscope* company for the official ceremony marking CERN's fiftieth anniversary, is a gift from the Government of the Republic and Canton of Geneva, which also wishes to share this moment of wonder with the local population. There will be three perfo...

  14. "Point de suspension"

    CERN Multimedia

    2004-01-01

    http://www.cern.ch/cern50/ CERN - Globe of Science and Innovation 20 and 21 October Acrobatics, mime, a cappella singing, projections of images, a magical setting... a host of different tools of a grandeur matching that of the Universe they relate. A camera makes a massive zoom out to reveal the multiple dimensions of Nature. Freeze the frame: half way between the infinitesimally small and the infinitesimally large, a man suspends his everyday life (hence the title "Point de Suspension", which refers to the three dots at the end of an uncompleted sentence) to take a glimpse of the place he occupies in the great history of the Universe. An unusual perspective on what it means to be a human being... This wondrous show in the Globe of Science and Innovation, specially created by the Miméscope* company for the official ceremony marking CERN's fiftieth anniversary, is a gift from the Government of the Republic and Canton of Geneva, which also wishes to share this moment of wonder with the local pop...

  15. "Point de suspension"

    CERN Document Server

    2004-01-01

    CERN - Globe of Science and Innovation 20 and 21 October Acrobatics, mime, a cappella singing, projections of images, a magical setting... a host of different tools of a grandeur matching that of the Universe they relate. A camera makes a massive zoom out to reveal the multiple dimensions of Nature. Freeze the frame: half way between the infinitesimally small and the infinitesimally large, a man suspends his everyday life (hence the title "Point de Suspension", which refers to the three dots at the end of an uncompleted sentence) to take a glimpse of the place he occupies in the great history of the Universe. An unusual perspective on what it means to be a human being... This spectacle in the Globe of Science and Innovation, specially created by the Miméscope* company for the official ceremony marking CERN's fiftieth anniversary, is a gift from the Government of the Republic and Canton of Geneva, which also wishes to share this moment of wonder with the local population. There will be three performances for...

  16. Simple and versatile turbidimetric monitoring of bacterial growth in liquid cultures using a customized 3D printed culture tube holder and a miniaturized spectrophotometer: application to facultative and strictly anaerobic bacteria

    Directory of Open Access Journals (Sweden)

    Margarida R. G. Maia

    2016-08-01

    Full Text Available Here we introduce a novel strategy for turbidimetric monitoring of bacterial growth in liquid culture. The instrumentation comprises a light source, a customized 3D printed culture tube holder and a miniaturized spectrophotometer, connected through optical cables. Due to its small footprint and the possibility to operate with external light, bacterial growth was directly monitored from culture tubes in a simple and versatile fashion. This new portable measurement technique was used to monitor the growth of facultative (Escherichia coli ATCC/25922, and Staphylococcus aureus ATCC/29213 and strictly (Butyrivibrio fibrisolvens JW11, Butyrivibrio proteoclasticus P18, and Propionibacterium acnes DSMZ 1897 anaerobic bacteria. For E. coli and S. aureus, the growth rates calculated from normalized optical density values were compared with those ones obtained using a benchtop spectrophotometer without significant differences (P = 0.256. For the strictly anaerobic species, a high precision (RSD < 3.5% was observed between replicates up to 48 h. Regarding its potential for customization, this manifold could accommodate further developments for customized turbidimetric monitoring, such as the use of light-emitting diodes as a light source or flow cells.

  17. Simple and Versatile Turbidimetric Monitoring of Bacterial Growth in Liquid Cultures Using a Customized 3D Printed Culture Tube Holder and a Miniaturized Spectrophotometer: Application to Facultative and Strictly Anaerobic Bacteria.

    Science.gov (United States)

    Maia, Margarida R G; Marques, Sara; Cabrita, Ana R J; Wallace, R John; Thompson, Gertrude; Fonseca, António J M; Oliveira, Hugo M

    2016-01-01

    Here we introduce a novel strategy for turbidimetric monitoring of bacterial growth in liquid culture. The instrumentation comprises a light source, a customized 3D printed culture tube holder and a miniaturized spectrophotometer, connected through optical cables. Due to its small footprint and the possibility to operate with external light, bacterial growth was directly monitored from culture tubes in a simple and versatile fashion. This new portable measurement technique was used to monitor the growth of facultative (Escherichia coli ATCC/25922, and Staphylococcus aureus ATCC/29213) and strictly (Butyrivibrio fibrisolvens JW11, Butyrivibrio proteoclasticus P18, and Propionibacterium acnes DSMZ 1897) anaerobic bacteria. For E. coli and S. aureus, the growth rates calculated from normalized optical density values were compared with those ones obtained using a benchtop spectrophotometer without significant differences (P = 0.256). For the strictly anaerobic species, a high precision (relative standard deviation < 3.5%) was observed between replicates up to 48 h. Regarding its potential for customization, this manifold could accommodate further developments for customized turbidimetric monitoring, such as the use of light-emitting diodes as a light source or flow cells.

  18. Preparation, characterisation and antibacterial activity of a florfenicol-loaded solid lipid nanoparticle suspension.

    Science.gov (United States)

    Wang, Ting; Chen, Xiaojin; Lu, Mengmeng; Li, Xihe; Zhou, WenZhong

    2015-12-01

    A florfenicol-loaded solid lipid nanoparticle (FFC-SLN) suspension was prepared by hot homogenisation and ultrasonic technique. The suspension was characterised for its release profile, stability, toxicity, and the physicochemical properties of the nanoparticles. Antibacterial activity of the suspension was evaluated in vitro and in vivo. The results showed that the mean diameter, polydispersity index and zeta potential of the nanoparticles were 253 ± 3 nm, 0.409 ± 0.022 and 47.5 ± 0.21 mV, respectively. In vitro release profile showed the FFC-SLN suspension had sustained release effect. The minimum inhibition concentration values of the FFC-SLN suspension were 6 and 3 µg/mL against Staphylococcus aureus and Escherichia coli respectively, compared with 3.5 and 2 µg/mL of native florfenicol. The suspension was relatively stable at 4°C and less stable at room temperature during 9 months storage. Although the nanoparticle carriers exhibited cytotoxicity in cell cultures, the LD50 of the lyophilised dry power of the suspension was higher than 5 g/kg body weight. Mortality protection against E. coli lethal infection in mice showed that the nanoparticle suspension had much better efficacy (6/10) than native drug (1/10). These results indicate that FFC-SLN suspension could be a promising formulation in veterinary medicine.

  19. Effects of transients in LIGO suspensions on searches for gravitational waves

    Science.gov (United States)

    Walker, M.; Abbott, T. D.; Aston, S. M.; González, G.; Macleod, D. M.; McIver, J.; Abbott, B. P.; Abbott, R.; Adams, C.; Adhikari, R. X.; Anderson, S. B.; Ananyeva, A.; Appert, S.; Arai, K.; Ballmer, S. W.; Barker, D.; Barr, B.; Barsotti, L.; Bartlett, J.; Bartos, I.; Batch, J. C.; Bell, A. S.; Betzwieser, J.; Billingsley, G.; Birch, J.; Biscans, S.; Biwer, C.; Blair, C. D.; Bork, R.; Brooks, A. F.; Ciani, G.; Clara, F.; Countryman, S. T.; Cowart, M. J.; Coyne, D. C.; Cumming, A.; Cunningham, L.; Danzmann, K.; Da Silva Costa, C. F.; Daw, E. J.; DeBra, D.; DeRosa, R. T.; DeSalvo, R.; Dooley, K. L.; Doravari, S.; Driggers, J. C.; Dwyer, S. E.; Effler, A.; Etzel, T.; Evans, M.; Evans, T. M.; Factourovich, M.; Fair, H.; Fernández Galiana, A.; Fisher, R. P.; Fritschel, P.; Frolov, V. V.; Fulda, P.; Fyffe, M.; Giaime, J. A.; Giardina, K. D.; Goetz, E.; Goetz, R.; Gras, S.; Gray, C.; Grote, H.; Gushwa, K. E.; Gustafson, E. K.; Gustafson, R.; Hall, E. D.; Hammond, G.; Hanks, J.; Hanson, J.; Hardwick, T.; Harry, G. M.; Heintze, M. C.; Heptonstall, A. W.; Hough, J.; Izumi, K.; Jones, R.; Kandhasamy, S.; Karki, S.; Kasprzack, M.; Kaufer, S.; Kawabe, K.; Kijbunchoo, N.; King, E. J.; King, P. J.; Kissel, J. S.; Korth, W. Z.; Kuehn, G.; Landry, M.; Lantz, B.; Lockerbie, N. A.; Lormand, M.; Lundgren, A. P.; MacInnis, M.; Márka, S.; Márka, Z.; Markosyan, A. S.; Maros, E.; Martin, I. W.; Martynov, D. V.; Mason, K.; Massinger, T. J.; Matichard, F.; Mavalvala, N.; McCarthy, R.; McClelland, D. E.; McCormick, S.; McIntyre, G.; Mendell, G.; Merilh, E. L.; Meyers, P. M.; Miller, J.; Mittleman, R.; Moreno, G.; Mueller, G.; Mullavey, A.; Munch, J.; Nuttall, L. K.; Oberling, J.; Oliver, M.; Oppermann, P.; Oram, Richard J.; O'Reilly, B.; Ottaway, D. J.; Overmier, H.; Palamos, J. R.; Paris, H. R.; Parker, W.; Pele, A.; Penn, S.; Phelps, M.; Pierro, V.; Pinto, I.; Principe, M.; Prokhorov, L. G.; Puncken, O.; Quetschke, V.; Quintero, E. A.; Raab, F. J.; Radkins, H.; Raffai, P.; Reid, S.; Reitze, D. H.; Robertson, N. A.; Rollins, J. G.; Roma, V. J.; Romie, J. H.; Rowan, S.; Ryan, K.; Sadecki, T.; Sanchez, E. J.; Sandberg, V.; Savage, R. L.; Schofield, R. M. S.; Sellers, D.; Shaddock, D. A.; Shaffer, T. J.; Shapiro, B.; Shawhan, P.; Shoemaker, D. H.; Sigg, D.; Slagmolen, B. J. J.; Smith, B.; Smith, J. R.; Sorazu, B.; Staley, A.; Strain, K. A.; Tanner, D. B.; Taylor, R.; Thomas, M.; Thomas, P.; Thorne, K. A.; Thrane, E.; Torrie, C. I.; Traylor, G.; Tuyenbayev, D.; Vajente, G.; Valdes, G.; van Veggel, A. A.; Vecchio, A.; Veitch, P. J.; Venkateswara, K.; Vo, T.; Vorvick, C.; Ward, R. L.; Warner, J.; Weaver, B.; Weiss, R.; Weßels, P.; Willke, B.; Wipf, C. C.; Worden, J.; Wu, G.; Yamamoto, H.; Yancey, C. C.; Yu, Hang; Yu, Haocun; Zhang, L.; Zucker, M. E.; Zweizig, J.

    2017-12-01

    This paper presents an analysis of the transient behavior of the Advanced LIGO (Laser Interferometer Gravitational-wave Observatory) suspensions used to seismically isolate the optics. We have characterized the transients in the longitudinal motion of the quadruple suspensions during Advanced LIGO's first observing run. Propagation of transients between stages is consistent with modeled transfer functions, such that transient motion originating at the top of the suspension chain is significantly reduced in amplitude at the test mass. We find that there are transients seen by the longitudinal motion monitors of quadruple suspensions, but they are not significantly correlated with transient motion above the noise floor in the gravitational wave strain data, and therefore do not present a dominant source of background noise in the searches for transient gravitational wave signals. Using the suspension transfer functions, we compared the transients in a week of gravitational wave strain data with transients from a quadruple suspension. Of the strain transients between 10 and 60 Hz, 84% are loud enough that they would have appeared above the sensor noise in the top stage quadruple suspension monitors if they had originated at that stage at the same frequencies. We find no significant temporal correlation with the suspension transients in that stage, so we can rule out suspension motion originating at the top stage as the cause of those transients. However, only 3.2% of the gravitational wave strain transients are loud enough that they would have been seen by the second stage suspension sensors, and none of them are above the sensor noise levels of the penultimate stage. Therefore, we cannot eliminate the possibility of transient noise in the detectors originating in the intermediate stages of the suspension below the sensing noise.

  20. Effects of transients in LIGO suspensions on searches for gravitational waves.

    Science.gov (United States)

    Walker, M; Abbott, T D; Aston, S M; González, G; Macleod, D M; McIver, J; Abbott, B P; Abbott, R; Adams, C; Adhikari, R X; Anderson, S B; Ananyeva, A; Appert, S; Arai, K; Ballmer, S W; Barker, D; Barr, B; Barsotti, L; Bartlett, J; Bartos, I; Batch, J C; Bell, A S; Betzwieser, J; Billingsley, G; Birch, J; Biscans, S; Biwer, C; Blair, C D; Bork, R; Brooks, A F; Ciani, G; Clara, F; Countryman, S T; Cowart, M J; Coyne, D C; Cumming, A; Cunningham, L; Danzmann, K; Da Silva Costa, C F; Daw, E J; DeBra, D; DeRosa, R T; DeSalvo, R; Dooley, K L; Doravari, S; Driggers, J C; Dwyer, S E; Effler, A; Etzel, T; Evans, M; Evans, T M; Factourovich, M; Fair, H; Fernández Galiana, A; Fisher, R P; Fritschel, P; Frolov, V V; Fulda, P; Fyffe, M; Giaime, J A; Giardina, K D; Goetz, E; Goetz, R; Gras, S; Gray, C; Grote, H; Gushwa, K E; Gustafson, E K; Gustafson, R; Hall, E D; Hammond, G; Hanks, J; Hanson, J; Hardwick, T; Harry, G M; Heintze, M C; Heptonstall, A W; Hough, J; Izumi, K; Jones, R; Kandhasamy, S; Karki, S; Kasprzack, M; Kaufer, S; Kawabe, K; Kijbunchoo, N; King, E J; King, P J; Kissel, J S; Korth, W Z; Kuehn, G; Landry, M; Lantz, B; Lockerbie, N A; Lormand, M; Lundgren, A P; MacInnis, M; Márka, S; Márka, Z; Markosyan, A S; Maros, E; Martin, I W; Martynov, D V; Mason, K; Massinger, T J; Matichard, F; Mavalvala, N; McCarthy, R; McClelland, D E; McCormick, S; McIntyre, G; Mendell, G; Merilh, E L; Meyers, P M; Miller, J; Mittleman, R; Moreno, G; Mueller, G; Mullavey, A; Munch, J; Nuttall, L K; Oberling, J; Oliver, M; Oppermann, P; Oram, Richard J; O'Reilly, B; Ottaway, D J; Overmier, H; Palamos, J R; Paris, H R; Parker, W; Pele, A; Penn, S; Phelps, M; Pierro, V; Pinto, I; Principe, M; Prokhorov, L G; Puncken, O; Quetschke, V; Quintero, E A; Raab, F J; Radkins, H; Raffai, P; Reid, S; Reitze, D H; Robertson, N A; Rollins, J G; Roma, V J; Romie, J H; Rowan, S; Ryan, K; Sadecki, T; Sanchez, E J; Sandberg, V; Savage, R L; Schofield, R M S; Sellers, D; Shaddock, D A; Shaffer, T J; Shapiro, B; Shawhan, P; Shoemaker, D H; Sigg, D; Slagmolen, B J J; Smith, B; Smith, J R; Sorazu, B; Staley, A; Strain, K A; Tanner, D B; Taylor, R; Thomas, M; Thomas, P; Thorne, K A; Thrane, E; Torrie, C I; Traylor, G; Tuyenbayev, D; Vajente, G; Valdes, G; van Veggel, A A; Vecchio, A; Veitch, P J; Venkateswara, K; Vo, T; Vorvick, C; Ward, R L; Warner, J; Weaver, B; Weiss, R; Weßels, P; Willke, B; Wipf, C C; Worden, J; Wu, G; Yamamoto, H; Yancey, C C; Yu, Hang; Yu, Haocun; Zhang, L; Zucker, M E; Zweizig, J

    2017-12-01

    This paper presents an analysis of the transient behavior of the Advanced LIGO (Laser Interferometer Gravitational-wave Observatory) suspensions used to seismically isolate the optics. We have characterized the transients in the longitudinal motion of the quadruple suspensions during Advanced LIGO's first observing run. Propagation of transients between stages is consistent with modeled transfer functions, such that transient motion originating at the top of the suspension chain is significantly reduced in amplitude at the test mass. We find that there are transients seen by the longitudinal motion monitors of quadruple suspensions, but they are not significantly correlated with transient motion above the noise floor in the gravitational wave strain data, and therefore do not present a dominant source of background noise in the searches for transient gravitational wave signals. Using the suspension transfer functions, we compared the transients in a week of gravitational wave strain data with transients from a quadruple suspension. Of the strain transients between 10 and 60 Hz, 84% are loud enough that they would have appeared above the sensor noise in the top stage quadruple suspension monitors if they had originated at that stage at the same frequencies. We find no significant temporal correlation with the suspension transients in that stage, so we can rule out suspension motion originating at the top stage as the cause of those transients. However, only 3.2% of the gravitational wave strain transients are loud enough that they would have been seen by the second stage suspension sensors, and none of them are above the sensor noise levels of the penultimate stage. Therefore, we cannot eliminate the possibility of transient noise in the detectors originating in the intermediate stages of the suspension below the sensing noise.

  1. Obstetric and perinatal outcomes of pregnancies conceived with embryos cultured in a time-lapse monitoring system.

    Science.gov (United States)

    Insua, Maria Fernanda; Cobo, Ana Cristina; Larreategui, Zaloa; Ferrando, Marcos; Serra, Vicente; Meseguer, Marcos

    2017-09-01

    To compare obstetric and perinatal outcomes of singleton pregnancies resulting from embryos incubated in a time-lapse system (TLS) with those of embryos grown in standard IVF incubators (SI). Retrospective description of a cohort of patients who conceived during a randomized, controlled trial. Private university-affiliated IVF center. Of 856 randomized patients, 378 gave birth to a live-born infant: 216 of the deliveries originated from embryos incubated in TLS, and 162 deliveries were from embryos cultured in SI. Embryo incubation and selection in TLS. Delivery and neonatal outcomes. No significant differences were observed in the baseline characteristics of the study population. The delivery rate was 49.3% (TLS) vs. 40.0% (SI), and multiple deliveries were higher in the TLS group: 31.0% (67 of 216) vs. 24.7% (40 of 162) in the SI group. When singleton pregnancies were analyzed no differences were found between the two groups in the rate of obstetric problems with respect to weeks at delivery: 38.8 (95% confidence interval [CI] 38.4-39.1) (TLS) vs. 39.5 (95% CI 38.0-39.9) (SI); preterm births (outcomes such as birth weight: 3,163 g (95% CI 3,035-3,292 g) (TLS) vs. 3,074 (95% CI 2,913-3,236) (SI); low birth weight (obstetric and perinatal outcomes when a time-lapse incubator was used rather than a more widely used conventional incubator. As far as we know this is the first report from a randomized study of the neonatal outcomes of time-lapse monitoring. Our results suggest that this technology is an effective and safe alternative for embryo incubation, though trials of larger numbers of patients are required to further confirm our conclusions. NCT01549262. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  2. Mixing Suspensions in Slender Tanks

    Directory of Open Access Journals (Sweden)

    F. Rieger

    2002-01-01

    Full Text Available Industrial suspension mixing processes are carried out both in standard tanks (H/D =1 and in the tanks with height H/D > 1. When only one impeller is used in such slender tanks, it may be difficult to produce a suspension of desired homogeneity. Hence it may be necessary to install a larger number of impellers on the shaft.The aim of this study was to explain the mechanism of suspension formation in slender tanks (H/D = 2 with an increased number of impellers. On the basis of the solid bed height on the tank bottom, the position of the suspension - water interface and the concentration profile of solid particles in the suspension (standard deviation of solid body concentration the operation of the impellers was estimated and conclusions were drawn on how and at what distance from each other to install them were presented.The location of the upper, highest impeller appeared to be specially significant. On the basis of this study it is recommended to locate the upper impeller so that its distance from the free liquid surface is less than 0.8 D. It was found that such a position of the highest impeller was also advantageous from the energy point of view.

  3. Comparison of techniques for the determination of conversion during suspension polymerization reactions

    Directory of Open Access Journals (Sweden)

    J. C. Santos

    2008-06-01

    Full Text Available The determination of conversion during suspension polymerization reactions is not an easy task due to the heterogeneity of the reaction medium and the tendency of particles to agglomerate rapidly when stirring is stopped. Usually, bulk polymerization in ampoules is employed to study the kinetics of suspension polymerization reactions. In this work, a comparison of different techniques for the determination of conversion during suspension polymerization reactions is presented. Results showed a good agreement between the conversion obtained by gravimetry during styrene suspension polymerization and on-line conversion monitoring data using fiber-optic based Raman Spectroscopy. Nevertheless, the polymerization rate of styrene bulk polymerization carried out in ampoules was higher than the real reaction rate of styrene suspension polymerization due to slightly higher reaction temperatures. Simulation results using the experimental temperature data in a mathematical model confirmed these results.

  4. Precision magnetic suspension linear bearing

    Science.gov (United States)

    Trumper, David L.; Queen, Michael A.

    1992-01-01

    We have shown the design and analyzed the electromechanics of a linear motor suitable for independently controlling two suspension degrees of freedom. This motor, at least on paper, meets the requirements for driving an X-Y stage of 10 Kg mass with about 4 m/sq sec acceleration, with travel of several hundred millimeters in X and Y, and with reasonable power dissipation. A conceptual design for such a stage is presented. The theoretical feasibility of linear and planar bearings using single or multiple magnetic suspension linear motors is demonstrated.

  5. Active Control of Suspension Bridges

    DEFF Research Database (Denmark)

    Thoft-Christensen, Palle

    In this paper some recent research on active control of very long suspension bridges, is presented. The presentation is based on research work at Aalborg University, Denmark. The active control system is based on movable flaps attached to the bridge girder. Wind load on bridges with or without...... flaps attached to the girder is briefly presented. A simple active control system is discussed. Results from wind tunnel experiments with a bridge section show that flaps can be used effectively to control bridge girder vibrations. Flutter conditions for suspension bridges with and without flaps...... are presented. The theory is illustrated by an example....

  6. Growth behavior in plant cell cultures based on emissions detected by a multisensor array.

    Science.gov (United States)

    Komaraiah, Palle; Navratil, Marian; Carlsson, Maria; Jeffers, Paul; Brodelius, Maria; Brodelius, Peter E; Kieran, Patricia M; Mandenius, Carl-Fredrik

    2004-01-01

    The use of a multisensor array based on chemical gas sensors to monitor plant cell cultures is described. The multisensor array, also referred to as an electronic nose, consisted of 19 different metal oxide semiconductor sensors and one carbon dioxide sensor. The device was used to continuously monitor the off-gas from two plant cell suspension cultures, Morinda citrifolia and Nicotiana tabacum, cultivated under batch conditions. By analyzing the multiarray responses using two pattern recognition methods, principal component analysis and artificial neural networks, it was possible to monitor the course of the cultivations and, in turn, to predict (1) the biomass concentration in both systems and (2) the formation of the secondary metabolite, antraquinone, by M. citrifolia. The results identify the multisensor array method as a potentially useful analytical tool for monitoring plant process variables that are otherwise difficult to analyze on-line.

  7. Relaxation of jammed colloidal suspensions after shear cessation.

    Science.gov (United States)

    Ianni, Francesca; Lasne, David; Sarcia, Régis; Hébraud, Pascal

    2006-07-01

    The dynamics of heterogeneities in a shear thickening, concentrated colloidal suspension is investigated through speckle visibility spectroscopy, a dynamic light scattering technique recently introduced [P. K. Dixon and D. J. Durian, Phys. Rev. Lett. 90, 184302 (2003)]. Formation of shear-induced heterogeneities is observed in the jamming regime, and their relaxation after shear cessation is monitored as a function of the applied shear stress. The relaxation time of these heterogeneities increases when a higher stress is applied.

  8. Effective temperature for sheared suspensions: A route towards closures for migration in bidisperse suspension

    NARCIS (Netherlands)

    Vollebregt, H.M.; Sman, van der R.G.M.

    2012-01-01

    By extending the concept of an effective temperature, earlier introduced for sheared monodisperse suspensions, we propose a continuum theory for sheared bidisperse suspensions. We show the theory for sheared suspensions can be constructed from the theory for Brownian suspensions by replacing the

  9. Rotational diffusion in dense suspensions

    NARCIS (Netherlands)

    Hagen, M. H. J.; Frenkel, D.; Lowe, C.P.

    1999-01-01

    We have computed the rotational diffusion coefficient for a suspension of hard spheres. We find excellent agreement with experimental results over a density range up to, and including, the colloidal crystal. However, we find that theories derived to second order in the volume fraction overestimate

  10. Suspension-Firing of Biomass

    DEFF Research Database (Denmark)

    Bashir, Muhammad Shafique; Jensen, Peter Arendt; Frandsen, Flemming

    2012-01-01

    This paper is Part 1 in a series of two describing probe measurements of deposit build-up and removal (shedding) in a 350 MWth suspension boiler, firing straw and wood. The influence of fuel type (straw share in wood), probe exposure time, probe surface temperature (500, 550, and 600 °C), and flu...

  11. Suspension-Firing of Biomass

    DEFF Research Database (Denmark)

    Shafique Bashir, Muhammad; Jensen, Peter Arendt; Frandsen, Flemming

    2012-01-01

    This paper is the second of two papers, describing probe measurements of deposit buildup and removal (shedding), conducted in a 350 MWth suspension-fired boiler, firing straw and wood. Investigations of deposit buildup and shedding have been made by use of an advanced online deposit probe and a s...

  12. Fracture in Kaolinite clay suspensions

    Science.gov (United States)

    Kosgodagan Acharige, Sebastien; Jerolmack, Douglas J.; Arratia, Paulo E.

    2017-11-01

    Clay minerals are involved in many natural (landslides, river channels) and industrial processes (ceramics, cosmetics, oil recovery). They are plate shaped charged colloids and exhibit different flow properties than simpler colloids when suspended in a liquid such as thixotropy and shear-banding. kaolinite platelets are non-swelling, meaning that the stacks formed by the platelets do not have water layers, and thus the suspension does not have a sol-gel transition. However, it has been shown that kaolinite suspensions possesses a non-zero yield stress even at low concentrations, indicating that the particles arrange themselves in a structure through attractive interactions. Here, we experimentally investigate the sedimentation of kaolinite suspensions in a Hele-Shaw cell. The sedimentation of these dilute suspensions can display solid behavior like fracture, revealed in cross-polarized light, which is linked to the failure of the weakly-bonded structure (typical yield stress 10-2 Pa). By changing the interaction potential of the particles (by sonication or introducing salts), we show through these sedimentation experiments, how the fracture pattern can be avoided. Research was sponsored by the Army Research Laboratory and was accomplished under Grant Number 569074.

  13. Nonlinear models of suspension bridges

    Czech Academy of Sciences Publication Activity Database

    Malík, Josef

    2006-01-01

    Roč. 321, č. 2 (2006), s. 828-850 ISSN 0022-247X Institutional research plan: CEZ:AV0Z30860518 Keywords : suspension bridges * principle of minimum energy Subject RIV: BA - General Mathematics Impact factor: 0.758, year: 2006

  14. Raman microspectroscopy: a noninvasive analysis tool for monitoring of collagen-containing extracellular matrix formation in a medium-throughput culture system.

    Science.gov (United States)

    Kunstar, Aliz; Otto, Cees; Karperien, Marcel; van Blitterswijk, Clemens; van Apeldoorn, Aart

    2011-07-01

    The three-dimensional environment is known to play an important role in promoting cell-matrix interactions. We have investigated the possibility of using Raman microspectroscopy--which has the great advantage of noninvasive sensing--for in vitro monitoring of extracellular matrix (ECM) formation in a medium-throughput pellet (3D) culture system with soft-litography, agarose-microwell arrays. Chondrocytes were seeded in the agarose microwells in basic or chondrocyte medium. After 3, 7, and 14 days of culture, samples were analyzed for ECM formation by Raman microspectroscopy, histology, and immunofluorescence. ECM formation in the chondrocyte medium-cultured samples was detected by histology and immunofluorescence, and also noninvasively by Raman microspectroscopy. The Raman band of collagen found at 937 cm(-1) can be used as a Raman marker for collagen-containing ECM formation over time in the chondrocyte pellets. This culture system can be implemented as a medium-throughput platform for Raman applications and screening microtissue formation, since with these agarose-microwell arrays relatively large numbers of cell pellets could be screened in a short time in situ, without having to transfer the pellets onto microscopic slides. Moreover, in this manner the culture system is suitable for long-term, real-time live-cell measurements.

  15. Intercomparison of Numerical Inversion Algorithms for Particle Size Determination of Polystyrene Suspensions Using Spectral Turbidimetry

    OpenAIRE

    Glasse, Benjamin; Riefler, Norbert; Fritsching, Udo

    2015-01-01

    The continuous monitoring of the particle size distribution in particulate processes with suspensions or emulsions requires measurement techniques that can be used as in situ devices in contrast to ex situ or laboratory methods. In this context, for the evaluation of turbidimetric spectral measurements, the application of different numerical inversion algorithms is investigated with respect to the particle size distribution determination of polystyrene suspensions. A modified regularization c...

  16. Dense suspensions: force response and jamming

    NARCIS (Netherlands)

    von Kann, S.

    2012-01-01

    The response of dense suspensions to an external force was studied using two different experiments. In the first experiment, objects were settled in a deep bath of a dense cornstarch suspension. This is the only suspension to result in two unexpected phenomena: Velocity oscillations in the bulk, and

  17. 49 CFR 570.8 - Suspension systems.

    Science.gov (United States)

    2010-10-01

    ... Pounds or Less § 570.8 Suspension systems. (a) Suspension condition. Ball joint seals shall not be cut or..., shall be installed on both front springs, both rear springs, or on all four springs. Shock absorber...) Inspection procedure. Examine front and rear end suspension parts for conditions indicated. (b) Shock...

  18. Alternatives to Suspensions: Rationale and Recommendations

    Science.gov (United States)

    Chin, Jenna K.; Dowdy, Erin; Jimerson, Shane R.; Rime, W. Jeremy

    2012-01-01

    Suspensions are often used as an individual disciplinary consequence in attempts to reduce problem behaviors in the future. However, suspensions have shown to be less effective for students with specific behavioral challenges and problems. When examining suspensions in the context of behaviorist and social-ecological learning theories, suspending…

  19. 49 CFR 570.61 - Suspension system.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 6 2010-10-01 2010-10-01 false Suspension system. 570.61 Section 570.61 Transportation Other Regulations Relating to Transportation (Continued) NATIONAL HIGHWAY TRAFFIC SAFETY... 10,000 Pounds § 570.61 Suspension system. (a) Suspension condition. Ball joint seals shall not be cut...

  20. Early detection of mycobacteria using a novel hydrogel culture method.

    Science.gov (United States)

    Jang, Mi Hee; Kim, Shine Young; Kim, Chang-Ki; Hwang, Sang-Hyun; Park, Byung Kyu; Kim, Sung Soo; Lee, Eun Yup; Chang, Chulhun L

    2014-01-01

    Early laboratory detection of Mycobacterium tuberculosis is crucial for controlling tuberculosis. We developed a hydrogel mycobacterial culture method that retains the advantages of both solid and liquid methods in terms of speed, cost, and efficiency. Mycobacterium bovis bacillus Calmette-Guérin (BCG) suspensions and 200 acid-fast bacilli (AFB)-positive clinical specimens were inoculated in Middlebrook 7H9 liquid media (Becton-Dickinson and Company, USA) and mixed with 75 µL of 9-fluorenylmethoxycarbonyl (Fmoc)-Phe-Phe-OH hydrogel stock solution in an Eppendorf tube just before culture incubation. The mixtures were cultured at 37℃ for as long as 14 days to monitor culture status. The number of M. bovis BCG increased with time. For 200 AFB smear-positive specimens, 155 of 158 conventional culture-positive specimens and 4 culture-negative or contaminated specimens yielded positive cultures within 14 days. For 128 specimens positive with the liquid culture method, the time to positive culture using the hydrogel method (mean, 12.6 days; range, 7 to 14 days) was significantly shorter than that for conventional liquid culture (mean, 16.2 days; range, 6 to 31 days; Pculture system is useful for timely, economical, and efficient detection of mycobacteria in clinical specimens.

  1. KINETIC DISTRIBUTION MODEL OF EVAPORATION, BIOSORPTION AND BIODEGRADATION OF POLYCHLORINATED BIPHENYLS (PCBS) IN THE SUSPENSION OF PSEUDOMONAS STUTZERI. (R826652)

    Science.gov (United States)

    AbstractKinetics of distribution of PCBs in an active bacterial suspension of Pseudomonas stutzeri was studied by monitoring the evaporated amounts and the concentration remaining in the liquid medium with the biomass. To determine the biodegradation rate const...

  2. Cytokine gene expression in murine epidermal cell suspensions: interleukin 1 beta and macrophage inflammatory protein 1 alpha are selectively expressed in Langerhans cells but are differentially regulated in culture.

    Science.gov (United States)

    Heufler, C; Topar, G; Koch, F; Trockenbacher, B; Kämpgen, E; Romani, N; Schuler, G

    1992-10-01

    Epidermal Langerhans cells (LC) are considered direct yet immature precursors of dendritic cells (DC) in the draining lymph nodes. Although the development of LC into potent immunostimulatory DC occurs in vitro and has been studied in detail, little is known about their profile of cytokine gene expression. By using reverse transcriptase polymerase chain reaction analysis to screen 16 cytokines followed by Northern blotting for selected analysis, we determined the cytokine gene expression profile of murine LC at different time points in culture when T cell stimulatory activity is increasing profoundly. LC regularly expressed macrophage inflammatory proteins, MIP-1 alpha and MIP-2, and interleukin 1 beta (IL-1 beta). Both MIPs were downregulated upon culture and maturation into DC, whereas IL-1 beta was strongly upregulated in culture. MIP-1 alpha and IL-1 beta mRNA were found only in LC, but not in other epidermal cells. Apart from trace amounts of IL-6 in cultured LC, several macrophage and T cell products were not detected. The cytokine expression profile of LC thus appears distinct from typical macrophages. The exact role of the cytokine genes we found transcribed in LC remains to be determined.

  3. A rational approach for the improvement of biomass production and lipid profile in cacao cell suspensions.

    Science.gov (United States)

    Rúa, Adriana María Gallego; Rojas, Luisa Fernanda; Trujillo, Aura Ines Urrea; Zuleta, Oriana Parra; Alvarez, Cristian David Correa; Garcés, Lucía Atehortúa

    2017-06-23

    Cocoa butter (CB) is produced in the seeds of Theobroma cacao representing 50% of its dry weight. The lipid composition plays an important role in the physicochemical, rheological, and sensory properties of the CB, making this fat a valuable resource for the production of chocolates, cosmetics, and pharmaceuticals. In this paper, are described experimental strategies used for a rational improvement of biomass production and fatty acids in cacao cell suspension cultures. First, the lipid profile in four cacao varieties is characterized, and then, one variety is selected to induce cell suspensions using a direct method without previous establishment of a callus phase. To improve growth and total fat production in cell suspension cultures, modified DKW media and newly designed media culture, based on the mineral concentrations of cacao seeds (cacao biomass production, "CBP"), are analyzed and compared. In addition, the effect of acetate in the lipid profile of cell suspensions is evaluated. Ultrastructural histological analysis of lipid vesicles in cacao seeds and cell suspensions is also performed. The results will show that it is feasible to establish cacao suspensions without the calli step and increase the biomass production by selecting a suitable cacao variety and tissue and also applying a new culture media formulation. In addition, it is possible to synthesize fatty acids in cell cultures and modify the lipid profile adding a precursor of the novo biosynthesis of fatty acids such as the acetate. Transmission electronic microscopy examinations and differential interference contrast microscopy analysis will demonstrate that lipid vesicles are the main reserve substance in both cacao seeds and cell suspensions.

  4. Suspensions with reduced violin string modes

    International Nuclear Information System (INIS)

    Lee, B H; Ju, L; Blair, D G

    2006-01-01

    We discuss the possibility of significantly reducing the number and Q-factor of violin string modes in the mirror suspension. Simulations of a bar-flexure suspension and an orthogonal ribbon have shown a reduction in the number of violin string modes when compared to a normal ribbon suspension. By calculating the expected suspension thermal noise, we find that the orthogonal ribbon provides a promising suspension alternative. A lower number of violin modes oscillating in the direction of the laser and a reduction in violin mode peak values of at least 23dB can be achieved with a slight increase in thermal noise above 40Hz

  5. Suspensions with reduced violin string modes

    Energy Technology Data Exchange (ETDEWEB)

    Lee, B H; Ju, L; Blair, D G [School of Physics, University of Western Australia, Crawley 6009, WA (Australia)

    2006-03-02

    We discuss the possibility of significantly reducing the number and Q-factor of violin string modes in the mirror suspension. Simulations of a bar-flexure suspension and an orthogonal ribbon have shown a reduction in the number of violin string modes when compared to a normal ribbon suspension. By calculating the expected suspension thermal noise, we find that the orthogonal ribbon provides a promising suspension alternative. A lower number of violin modes oscillating in the direction of the laser and a reduction in violin mode peak values of at least 23dB can be achieved with a slight increase in thermal noise above 40Hz.

  6. Suspension state increases reattachment of breast cancer cells by up-regulating lamin A/C.

    Science.gov (United States)

    Zhang, Xiaomei; Lv, Yonggang

    2017-12-01

    Extravasation is a rate-limiting step of tumor metastasis, for which adhesion to endothelium of circulating tumor cells (CTCs) is the prerequisite. The suspension state of CTCs undergoing detachment from primary tumor is a persistent biomechanical cue, which potentially regulates the biophysical characteristics and cellular behaviors of tumor cells. In this study, breast tumor cells MDA-MB-231 in suspension culture condition were used to investigate the effect of suspension state on reattachment of CTCs. Our study demonstrated that suspension state significantly increased the adhesion ability of breast tumor cells. In addition, suspension state markedly promoted the formation of stress fibers and focal adhesions and reduced the motility in reattached breast cancer cells. Moreover, lamin A/C was reversibly accumulated at posttranscriptional level under suspension state, improving the cell stiffness of reattached breast cancer cells. Disruption of actin cytoskeleton by cytochalasin D caused lamin A/C accumulation. Conversely, decreasing actomyosin contraction by ROCK inhibitor Y27632 reduced lamin A/C level. Knocking down lamin A/C weakened the suspension-induced increase of adhesion, and also abolished the suspension-induced decrease of motility and increase of stress fibers and focal adhesion in reattaching tumor cells, suggesting a crucial role of lamin A/C. In conclusion, it was demonstrated that suspension state promoted the reattachment of breast tumor cells by up-regulating lamin A/C via cytoskeleton disruption. These findings highlight the important role of suspension state for tumor cells in tumor metastasis. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. ON-LINE MONITORING OF BIOMASS CONCENTRATION BASED ON A CAPACITANCE SENSOR: ASSESSING THE METHODOLOGY FOR DIFFERENT BACTERIA AND YEAST HIGH CELL DENSITY FED-BATCH CULTURES

    Directory of Open Access Journals (Sweden)

    A. C. L. Horta

    2015-12-01

    Full Text Available Abstract The performance of an in-situ capacitance sensor for on-line monitoring of biomass concentration was evaluated for some of the most important microorganisms in the biotechnology industry: Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris and Bacillus megaterium. A total of 33 batch and fed-batch cultures were carried out in a bench-scale bioreactor and biomass formation trends were followed by dielectric measurements during the growth phase as well as the induction phase, for 5 recombinant E. coli strains. Permittivity measurements and viable cellular concentrations presented a linear correlation for all the studied conditions. In addition, the permittivity signal was further used for inference of the cellular growth rate. The estimated specific growth rates mirrored the main trends of the metabolic states of the different cells and they can be further used for setting-up control strategies in fed-batch cultures.

  8. Antioxidant isoenzyme responses to nickel-induced stress in tobacco cell suspension culture Resposta de isoenzimas antioxidantes ao estresse induzido por níquel em cultura de células em suspensão de fumo

    Directory of Open Access Journals (Sweden)

    Georgia Bertoni Pompeu

    2008-01-01

    Full Text Available Exposure to nickel (Ni at high concentrations can lead to production of reactive oxygen species (ROS resulting in oxidative damage at the cellular level. We investigated the antioxidative responses of Nicotiana tabacum cv BY-2 cell suspension to Ni stress (0.075 and 0.75 mM NiCl2 over a 72 h period with special attention to potential alterations in isoenzymes of superoxide dismutase (SOD, catalase (CAT and glutathione reductase (GR. Two main SOD isoenzymes were observed, a Mn-SOD (band I and a Fe-SOD (band II, as well as one CAT isoenzyme and four GR isoenzymes. Activity staining analysis revealed that CAT activity plays a major role in the early response to Ni-induced oxidative stress, particularly when the Ni concentration used was low, whilst a specific GR isoenzyme appears to respond to the Ni-induced oxidative stress when a much higher Ni concentration was used to induce the stress for the same period of treatment. These results illustrate the importance and advantages of determining individual isoenzyme activities.A exposição ao níquel (Ni, em altas concentrações, pode levar à produção de espécies reativas de oxigênio (EAOs, resultando em danos oxidativos em nível celular. Foram investigadas as respostas antioxidativas de células em suspensão do cultivar BY-2 de Nicotiana tabacum submetidas ao estresse por Ni (0.075 e 0.75 mM de NiCl2 por 72 h, com atenção especial às alterações potencias em isoenzimas de superóxido dismutase (SOD, catalase (CAT e glutatione redutase (GR. Duas principais isoenzimas de SOD foram observadas, uma Mn-SOD (banda I e outra Fe-SOD (banda II, bem como uma isoenzima CAT e quatro isoenzimas de GR. As análises revelaram que a atividade de CAT tem papel principal no momento inicial de resposta ao estresse oxidativo induzido por Ni, particularmente, quando sua concentração foi mais baixa, enquanto uma isoenzima específica de GR parece responder a este estresse na concentração mais alta de Ni

  9. Robust Tensioned Kevlar Suspension Design

    Science.gov (United States)

    Young, Joseph B.; Naylor, Bret J.; Holmes, Warren A.

    2012-01-01

    One common but challenging problem in cryogenic engineering is to produce a mount that has excellent thermal isolation but is also rigid. Such mounts can be achieved by suspending the load from a network of fibers or strings held in tension. Kevlar fibers are often used for this purpose owing to their high strength and low thermal conductivity. A suite of compact design elements has been developed to improve the reliability of suspension systems made of Kevlar.

  10. Particle Suspension Mechanisms - Supplemental Material

    Energy Technology Data Exchange (ETDEWEB)

    Dillon, M B

    2011-03-03

    This supplemental material provides a brief introduction to particle suspension mechanisms that cause exfoliated skin cells to become and remain airborne. The material presented here provides additional context to the primary manuscript and serves as background for designing possible future studies to assess the impact of skin cells as a source of infectious aerosols. This introduction is not intended to be comprehensive and interested readers are encouraged to consult the references cited.

  11. Monitoring of Vibrations for the Conservation of Cultural Heritage the Case of the Summer Palace in Beijing

    Science.gov (United States)

    Gao, X.; Zhang, L.; Liu, G.; Ba, Z.

    2017-08-01

    In order to research the influence of the environmental vibration and the protection of the architectural heritage, CMG-5TCDE accelerometer was used to obtain the real-time vibration monitoring data of the Grand Stage in the Garden of Virtuous Harmony in Summer Palace. This paper employs the control variable method and the multivariate statistical analysis, integrating monitoring statistics with wind speed and transportation observation statistics to make interrelated analysis, in order to study the response rules and impact degree of the Grand Stage under the environmental vibration impact. The analyses of the vibration monitoring statistic of the recent two years show that the vibration acceleration of the Grand Stage is below 10mm/s2, which is remarkable related with factors such as visitors flow rate, transportation and weather.

  12. Polymorphism in Bacterial Flagella Suspensions

    Science.gov (United States)

    Schwenger, Walter J.

    Bacterial flagella are a type of biological polymer studied for its role in bacterial motility and the polymorphic transitions undertaken to facilitate the run and tumble behavior. The naturally rigid, helical shape of flagella gives rise to novel colloidal dynamics and material properties. This thesis studies methods in which the shape of bacterial flagella can be controlled using in vitro methods and the changes the shape of the flagella have on both single particle dynamics and bulk material properties. We observe individual flagellum in both the dilute and semidilute regimes to observe the effects of solvent condition on the shape of the filament as well as the effect the filament morphology has on reptation through a network of flagella. In addition, we present rheological measurements showing how the shape of filaments effects the bulk material properties of flagellar suspensions. We find that the individual particle dynamics in suspensions of flagella can vary with geometry from needing to reptate linearly via rotation for helical filaments to the prevention of long range diffusion for block copolymer filaments. Similarly, for bulk material properties of flagella suspensions, helical geometries show a dramatic enhancement in elasticity over straight filaments while block copolymers form an elastic gel without the aid of crosslinking agents.

  13. Monitoring UV-induced signalling pathways in an ex vivo skin organ culture model using phospho-antibody array.

    Science.gov (United States)

    Lenain, Christelle; Gamboa, Bastien; Perrin, Agnes; Séraïdaris, Alexia; Bertino, Béatrice; Rival, Yves; Bernardi, Mathieu; Piwnica, David; Méhul, Bruno

    2017-09-08

    We investigated UV-induced signalling in an ex vivo skin organ culture model using phospho-antibody array. Phosphorylation modulations were analysed in time-course experiments following exposure to solar-simulated UV and validated by Western blot analyses. We found that UV induced P-p38 and its substrates, P-ERK1/2 and P-AKT, which were previously shown to be upregulated by UV in cultured keratinocytes and in vivo human skin. This indicates that phospho-antibody array applied to ex vivo skin organ culture is a relevant experimental system to investigate signalling events following perturbations. As the identified proteins are components of pathways implicated in skin tumorigenesis, UV-exposed skin organ culture model could be used to investigate the effect on these pathways of NMSC cancer drug candidates. In addition, we found that phospho-HCK is induced upon UV exposure, producing a new candidate for future studies investigating its role in the skin response to UV and UV-induced carcinogenesis. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. A procedure for monitoring the presence of the virulence plasmid (pYV) in Yersinia pestis under culture conditions

    Science.gov (United States)

    The pathogenicity of Yersinia pestis depends on the presence of a virulence plasmid (pYV). The unstable nature of pYV in Y. pestis leads to the eventual outgrowth of pYV less cells due its higher growth rate. Thus, it was necessary to develop procedures to monitor the presence of the plasmid durin...

  15. Melding the Old with the New: Trends in Methods Used to Identify, Monitor, and Control Microorganisms on Cultural Heritage Materials.

    Science.gov (United States)

    Sanmartín, Patricia; DeAraujo, Alice; Vasanthakumar, Archana

    2016-04-27

    Microbial activity has an important impact on the maintenance of cultural heritage materials, owing to the key role of microorganisms in many deterioration processes. In order to minimize such deleterious effects, there is a need to fine-tune methods that detect and characterize microorganisms. Trends in microbiology indicate that this need can be met by incorporating modern techniques. All of the methods considered in this review paper are employed in the identification, surveillance, and control of microorganisms, and they have two points in common: They are currently used in microbial ecology (only literature from 2009 to 2015 is included), and they are often applied in the cultural heritage sector. More than 75 peer-reviewed journal articles addressing three different approaches were considered: molecular, sensory and morphological, and biocontrol methods. The goal of this review is to highlight the usefulness of the traditional as well as the modern methods. The general theme in the literature cited suggests using an integrated approach.

  16. Culture-dependent and independent techniques to monitor yeast species during cold soak carried out at different temperatures in winemaking.

    Science.gov (United States)

    Maturano, Y Paola; Mestre, M Victoria; Combina, Mariana; Toro, María Eugenia; Vazquez, Fabio; Esteve-Zarzoso, Braulio

    2016-11-21

    Transformation of grape must into wine is a process that may vary according to the consumers' requirements. Application of cold soak prior to alcoholic fermentation is a common practice in cellars in order to enhance flavor complexity and extraction of phenolic compounds. However, the effect of this step on wine yeast microbiota is not well-known. The current study simultaneously analyzed the effect of different cold soak temperatures on the microbiological population throughout the process and the use of culture-dependent and independent techniques to study this yeast ecology. The temperatures assayed were those normally applied in wineries: 2.5, 8 and 12°C. PCR-DGGE allowed detection of the most representative species such as Hanseniaspora uvarum, Starmerella bacillaris and Saccharomyces cerevisiae. As could be expected, highest diversity indices were obtained at the beginning of each process, and survival of H. uvarum or S. bacillaris depended on the temperature. Our results are in agreement with those obtained with culture independent methods, but qPCR showed higher precision and a different behavior was observed for each yeast species and at each temperature assayed. Comparison of both culture-independent techniques can provide a general overview of the whole process, although DGGE does not reveal the diversity expected due to the reported problems with the sensitivity of this technique. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Comparison between xCELLigence biosensor technology and conventional cell culture system for real-time monitoring human tenocytes proliferation and drugs cytotoxicity screening.

    Science.gov (United States)

    Chiu, Chih-Hao; Lei, Kin Fong; Yeh, Wen-Ling; Chen, Poyu; Chan, Yi-Sheng; Hsu, Kuo-Yao; Chen, Alvin Chao-Yu

    2017-10-16

    tenocytes could proliferate inside xCELLigence system. These responses varied when tenocytes were exposed to different concentrations of ketorolac tromethamine, bupivacaine, methylprednisolone, and betamethasone. The result of cell proliferation and gene expression of tenocytes in both xCELLigence and conventional culture system is strongly correlated. xCELLigence culture system may replace conventional cell culture, which made real-time tenocyte proliferation monitoring possible.

  18. Monitoring of lactic fermentation driven by different starter cultures via direct injection mass spectrometric analysis of flavour-related volatile compounds.

    Science.gov (United States)

    Benozzi, Elisabetta; Romano, Andrea; Capozzi, Vittorio; Makhoul, Salim; Cappellin, Luca; Khomenko, Iuliia; Aprea, Eugenio; Scampicchio, Matteo; Spano, Giuseppe; Märk, Tilmann D; Gasperi, Flavia; Biasioli, Franco

    2015-10-01

    In this work, we used Proton Transfer Reaction-Mass Spectrometry (PTR-ToF-MS), coupled with an automated sampling system, to monitor lactic fermentation driven by different yogurt commercial starter cultures via direct injection mass spectrometric analysis of flavour-related volatile compounds. The aim is the identification of markers for real-time and non-invasive bioprocess control and optimisation as an industrial driver of innovation in food technology and biotechnology. We detected more than 300 mass peaks, tentatively identifying all major yogurt aroma volatiles. Thirteen mass peaks showed statistically significant differences among the four commercial starters. Among these are acetaldehyde, methanethiol, butanoic acid, 2-butanone, diacetyl, acetoin, 2-hydroxy-3-pentanone/pentanoic acid, heptanoic acid and benzaldehyde which play a key role in yogurt flavour. These volatile described the diverse flavour properties claimed by food biotechnological companies and, considering the possible contribution to yogurt flavour, are potential markers for the rapid screening of starter cultures and for the quality design in this fermentation-driven production. The strength of our approach lies in the identification, for the first time, of specific depletion kinetics of four sulphur containing compounds occurring during fermentation (hydrogen sulphide, methanethiol, S-methyl thioacetate/S-ethyl thioformate, pentane-thiol), which suggest a new possible protechnological feature of yogurt starter cultures. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Time Varying Behavior of the Loudspeaker Suspension

    DEFF Research Database (Denmark)

    Pedersen, Bo Rohde; Agerkvist, Finn T.

    2007-01-01

    The suspension part of the electrodynamic loudspeaker is often modelled as a simple linear spring with viscous damping, however the dynamic behaviour of the suspension is much more complicated than predicted by such a simple model. At higher levels the compliance becomes non-linear and often chan...... changes during excitation at high levels. This paper investigates how the compliance of the suspension depends on the excitation, i.e. level and frequency content. The measurements are compared with other known measurement methods of the suspension....

  20. Monitoring techniques: “StegoGIS: a geographical information system for knowing and preventing infestation risks in cultural heritage”

    Directory of Open Access Journals (Sweden)

    Katia Baslé

    2011-08-01

    Full Text Available Since 2004, the Cicrp: “Centre Interrégional de Conservation et Restauration du Patrimoine”, located in Marseilles has been involved in an interdisciplinary research program dealing with infestation and re-infestation, on lining pastes used in painting conservation, by the Stegobium paniceum through a GIS system : a geographical information system called “StegoGIS”. The GIS helps understand the insects ethology in its environmental context, mainly in flour and semolina millings in order to determine analysis criteria to prevent, mitigate and fight infestation in the cultural property environment. Our approach is based on three main lines: 1- A transverse approach of infestation in any type of cultural heritage institution: archives, libraries, museums, historic buildings where organic material collections and environment are attractive on an “insect point of view”. 2 –An IPM strategy (Integrated Pest Management including conservation and management of collections and buildings also based on an infestation survey with actual or potential risks. 3- A “decision- making tool” in diagnosis, preventing methods and treatments for professional conservation staff.

  1. Design analysis of formula student race car suspension system

    Science.gov (United States)

    Wirawan, Julian Wisnu; Ubaidillah, Aditra, Rama; Alnursyah, Rafli; Rahman, Rizki Abdul; Cahyono, Sukmaji Indro

    2018-02-01

    Design analysis of suspension especially for racecar suspension is very crucial to achieve maximum performance and handling. Suspension design may vary depending on the road terrain and the vehicle purpose itself, such as high speed or off-road vehicle. This paper focused on the suspension which used for racecar vehicle. The suspension type used was unequal double wishbone. This model is used because of its stability for high-speed usage compared to another kind of suspension. The suspension parameter was calculated to achieve desired performance. The result is the motion ratio of the designed suspension geometry. The obtained value of motion ratio was 1:2 for front suspension and 1:1 for the rear suspension. These calculation result the front suspension is still too soft, which the optimal motion ratio should be kept around 1:1 for better handling. This problem caused by the lack of space for suspension linkage.

  2. 39 CFR 320.9 - Revocation or amendment of suspensions.

    Science.gov (United States)

    2010-07-01

    ... SUSPENSION OF THE PRIVATE EXPRESS STATUTES § 320.9 Revocation or amendment of suspensions. These suspensions... of operations (in dollar or volume terms, whichever is larger) lower than that antedating the...

  3. Photoelectrochemical studies on aqueous suspensions of some ...

    Indian Academy of Sciences (India)

    Keywords. Photoelectrochemical; nanomaterial; suspensions; hydrogen; oxides; chalcogenides. Abstract. Photoproduction of hydrogen was achieved by photolysis of aqueous suspensions of mixed TiO2/V2O5 or CdS/ZnS semiconductor (SC) nanoparticle in phosphate buffers containing [Fe(CN)6]4−. Manipulations of the ...

  4. Alternatives to Suspension: A Government Initiative

    Science.gov (United States)

    Ryan, Thomas G.; Zoldy, Scott

    2011-01-01

    Student discipline and the ineffectiveness of out-of-school suspension is examined in light of the Ontario (Canada) legislative reform that supported a greater emphasis on progressive discipline alternatives to out-of-school suspension. Alternative discipline herein is explored via the behavior education plan, the school survival group, and…

  5. 78 FR 54766 - Suspension of Community Eligibility

    Science.gov (United States)

    2013-09-06

    ... (NFIP) that are scheduled for suspension on the effective dates listed within this rule because of... Status Book (CSB). The CSB is available at http://www.fema.gov/fema/csb.shtm . DATES: Effective Dates... suspension date. Since these notifications were made, this final rule may take effect within less than 30...

  6. 49 CFR 238.227 - Suspension system.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Suspension system. 238.227 Section 238.227 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION... Equipment § 238.227 Suspension system. On or after November 8, 1999— (a) All passenger equipment shall...

  7. 36 CFR 25.3 - Supervision; suspensions.

    Science.gov (United States)

    2010-07-01

    ... 36 Parks, Forests, and Public Property 1 2010-07-01 2010-07-01 false Supervision; suspensions. 25.3 Section 25.3 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR NATIONAL MILITARY PARKS; LICENSED GUIDE SERVICE REGULATIONS § 25.3 Supervision; suspensions. (a) The guide...

  8. On the advance of non-invasive techniques implementation for monitoring moisture distribution in cultural heritage: a case study

    Science.gov (United States)

    Inmaculada Martínez Garrido, María; Gómez Heras, Miguel; Fort González, Rafael; Valles Iriso, Javier; José Varas Muriel, María

    2015-04-01

    This work presents a case study developed in San Juan Bautista church in Talamanca de Jarama (12th -16th Century), which have been selected as an example of a historical church with a complex construction with subsequent combination of architectural styles and building techniques and materials. These materials have a differential behavior under the influence of external climatic conditions and constructive facts. Many decay processes related to humidity are affecting the building's walls and also have influence in the environmental dynamics inside the building. A methodology for monitoring moisture distribution on stone and masonry walls and floors was performed with different non-invasive techniques as thermal imaging, wireless sensor networks (WSN), portable moisture meter, electrical resistivity tomography (ERT) and ground-penetrating radar (GPR), in order to the evaluate the effectiveness of these techniques for the knowledge of moisture distribution inside the walls and the humidity origin. North and south oriented sections, both on walls and floors, were evaluated and also a general inspection in the church was carried out with different non-invasive techniques. This methodology implies different monitoring stages for a complete knowledge of the implication of outdoors and indoors conditions on the moisture distribution. Each technique is evaluated according to its effectiveness in the detection of decay processes and maintenance costs. Research funded by Geomateriales (S2013/MIT-2914) and Deterioration of stone materials in the interior of historic buildings as a result induced variation of its microclimate (CGL2011-27902) projects. The cooperation received from the Complutense University of Madrid's Research Group Alteración y Conservación de los Materiales Pétreos del Patrimonio (ref. 921349), the Laboratory Network in Science and Technology for Heritage Conservation (RedLabPat, CEI Moncloa) and the Diocese of Alcalá is gratefully acknowledged. MI Mart

  9. Use of molecular methods to characterize the bacterial community and to monitor different native starter cultures throughout the ripening of Galician chorizo.

    Science.gov (United States)

    Fonseca, Sonia; Ivette Ouoba, Labia Irène; Franco, Inmaculada; Carballo, Javier

    2013-05-01

    The development of Lactobacillus and Staphylococcus strains used as starter cultures throughout the ripening of Galician chorizo, a traditional dry fermented sausage from the north-west of Spain, was monitored combining different molecular-based techniques. The bacterial diversity occurring in the inoculated sausages at the beginning and the end of the ripening was also studied and compared to the indigenous population in an uninoculated control batch. Real-time PCR was used to monitor the Lactobacillus and Staphylococcus community using genus and species-specific primer to quantify the occurring microbiota. The identification of isolates at genus or species level was achieved by specific PCR and 16S rRNA gene sequencing. rep-PCR using (GTG)5-PCR primer was used to characterize this bacterial community at strain level. According to the data obtained, the strains Lactobacillus sakei LS131, Staphylococcus equorum SA25 and Staphylococcus saprophyticus SB12 were dominant during the ripening process, whereas the strain Staphylococcus epidermidis SA49, that was added in order to study its behaviour with a merely scientific purpose, did not succeed in dominating ripening, since it seemed to be outcompeted by autochthonous microbiota. In conclusion, the combination of a quantitative method such as real-time PCR with the identification and typing techniques used in this study (genus and species-specific PCR, 16S rRNA gene sequencing and (GTG)5-PCR) provided accurate and complete information about the starter cultures development, assessing their growth and survival over the ripening process. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Osmotic consolidation of suspensions and gels

    International Nuclear Information System (INIS)

    Miller, K.T.; Zukoski, C.F.

    1994-01-01

    An osmotic method for the consolidation of suspensions of ceramic particles is demonstrated. Concentrated solutions of poly(ethylene oxide) are separated from a suspension of ceramic particles by a semipermeable membrane, creating a gradient in solvent chemical potential. Solvent passes from the suspension into the polymer solution, lowering its free energy and consolidating the suspension. Dispersions of stable 8-nm hydrous zirconia particles were consolidated to over 47% by volume. Suspensions of α-alumina in three states of aggregation (dispersed, weakly flocculated, and strongly flocculated) were consolidated to densities greater than or equal to those produced in conventional pressure filtration. Moreover, the as-consolidated alumina bodies were partially drained of fluid during the osmotic consolidation process, producing cohesive partially dried bodies with improved handling characteristics

  11. Research of Biogas Purification Using Microalgae Monoraphidium Griffithii Suspension

    Directory of Open Access Journals (Sweden)

    Živilė Bingelytė

    2017-09-01

    Full Text Available Using biogas instead of fossil fuels decreases pollutants such as solid particles, sulphur dioxide, nitrogen oxides concentrations in the environment. Green energy and development of relevant infrastructure improves air quality considerably. Chemical, physical, biological methods are used for biogas purification. The main difficulties using biological methods are selection of suitable microorganisms’ suspensions and making optimal conditions in photobioreactor. Different origin and structure microalgae suspensions are used applying biological treatment methods. Monoraphidium griffithi, which is widespread in fresh water, has relatively high potential. Microalgae’ cultures absorb the main components of biogas – carbon dioxide (CO2 and hydrogen sulphide (H2S. Absorbtion processes are based on photosynthesis. Microalgae absorb specific components of biogas when there are suitable light source and nutrient solvent. The main purposes of the research are to asses emission of biogas using different substrates (chicken manure and wastewater sludge. Also, it was measured main physical and chemical characteristics of both substrates: acidicy, temperature, redox potential, conductivity, biohemical oxygen demand. According results of the research, emission from wastewater sludge is greater than from chicken manure so sludge was chosen in teh next stage of the research. The next stage – asssessment of purification efficienty using Monoraphidium grifftihii suspension. Raw biogas was supplied to photobioreacor (with microalgae suspension. Alterations of methane, carbon dioxide, oxygen, hydrogen sulphide concentrations were measured precisely. According to results concentration of methane in the beginning of the researc was 62%, after 35 days – 69%. Meanwhile carbon dioxide – 37% and 31% by analogy. Experimental research alows to assess Monoraphidium griffithi absorption capacity of ballast components. Results were compared to different scientists

  12. Development and application of a functional CE-SSCP fingerprinting method based on [Fe-Fe]-hydrogenase genes for monitoring hydrogen-producing Clostridium in mixed cultures

    Energy Technology Data Exchange (ETDEWEB)

    Quemeneur, Marianne; Hamelin, Jerome; Latrille, Eric; Steyer, Jean-Philippe; Trably, Eric [INRA, UR050, Laboratoire de Biotechnologie de l' Environnement, avenue des Etangs, Narbonne F-11100 (France)

    2010-12-15

    A Capillary Electrophoresis Single Strand Conformation Polymorphism (CE-SSCP) method based on functional [Fe-Fe]-hydrogenase genes was developed for monitoring the hydrogen (H{sub 2})-producing clostridial population in mixed-culture bioprocesses. New non-degenerated primers were designed and then validated on their specific PCR detection of a broad range of clostridial hydA genes. The hydA-based CE-SSCP method gave a specific and discriminating profile for each of the Clostridium strains tested. This method was validated using H{sub 2}-producing mixed cultures incubated at temperatures ranging from 25 C to 45 C. The hydA CE-SSCP profiles clearly differed between temperatures tested. Hence, they varied according to variations of the H{sub 2} production performances. The HydA sequences amplified with the new primer set indicated that diverse Clostridium strains impacted the H{sub 2} production yields. The highest performances were related to the dominance of Clostridium sporogenes-like hydA sequences. This CE-SSCP tool offers highly reliable and throughput analysis of the functional diversity and structure of the hydA genes for better understanding of the H{sub 2}-producing clostridial population dynamics in H{sub 2} dark fermentation bioreactors. (author)

  13. [Comparison of two types of cell cultures for preparation of sTNFRII-gAD fusion protein].

    Science.gov (United States)

    Huang, Shigao; Yin, Yuting; Xiong, Chunhui; Wang, Caihong; Lü, Jianxin; Gao, Jimin

    2013-01-01

    In this study we used two types of cell cultures, i.e., anchorage-dependent basket and full suspension batch cultures of sTNFRII-gAD-expressing CHO cells in the CelliGen 310 bioreactor (7.5 L) to compare their yields in order to optimize the culturing conditions for efficient expression of sTNFRII-gAD fusion protein consisting of soluble tumor necrosis factor receptor II and globular domain of adiponectin. The anchorage-dependent basket culture was performed in 4L 10% serum-containing medium with the final inoculating concentration of 3 x 10(5) to 4 x 10(5) cells/mL of sTNFRII-gAD-expressing CHO cells for 3 days, and then switched to 4 L serum-free LK021 medium to continue the culture for 4 days. The full suspension batch culture was carried out in the 4 L serum-free LK021 medium with the final inoculating concentration of 3 x 10(5) to 4 x 10(5) cells/mL of sTNFRII-gAD-expressing CHO cells for 7 days. The culturing conditions were monitored in real-time to maintain pH and dissolved oxygen stability through the whole process. The supernatants were collected by centrifuge, and the protein was concentrated through Pellicon flow ultrafiltration system and then purified by DEAE anion exchange. The results showed that the yields of sTNFRII-gAD fusion protein were 8.0 mg/L with 95% purity and 7.5 mg/L with 98% purity in the anchorage-dependent basket and the full suspension batch cultures, respectively. The study provided the framework for the pilot production of sTNFRII-gAD fusion protein.

  14. Nanometer-scale sizing accuracy of particle suspensions on an unmodified cell phone using elastic light scattering.

    Directory of Open Access Journals (Sweden)

    Zachary J Smith

    Full Text Available We report on the construction of a Fourier plane imaging system attached to a cell phone. By illuminating particle suspensions with a collimated beam from an inexpensive diode laser, angularly resolved scattering patterns are imaged by the phone's camera. Analyzing these patterns with Mie theory results in predictions of size distributions of the particles in suspension. Despite using consumer grade electronics, we extracted size distributions of sphere suspensions with better than 20 nm accuracy in determining the mean size. We also show results from milk, yeast, and blood cells. Performing these measurements on a portable device presents opportunities for field-testing of food quality, process monitoring, and medical diagnosis.

  15. Changes in gene expression during programmed cell death in tomato cell suspensions

    NARCIS (Netherlands)

    Hoeberichts, F.A.; Orzaez, D.; Plas, van der L.H.W.; Woltering, E.J.

    2001-01-01

    To identify genes involved in plant programmed cell death (PCD), changes in gene expression during PCD in a model system of suspension-cultured tomato cells were studied. In this system, cell death is triggered by treatment with camptothecin, an inhibitor of topoisomerase I. Cell death was

  16. [Biotransformation of daphnetin by suspension transgenic hairy roots of Polygonum multiflorum].

    Science.gov (United States)

    Deng, Wenjuan; Zhou, Liangbin; Yu, Rongmin

    2011-02-01

    To investigate the biotransformation of daphnetin by suspension transgenic hairy root of Polygonum multiflorum and provide a biotechnological method for large-scale production of the daphnetin-8-O-beta-D-glucoside using this new culture system. Daphnetin was added into the media of suspension to culture 36 h. The biotransformation product was detected with TLC and HPLC, and isolated by various chromatographic methods. The influence of co-cultured time on conversion ratio, content of degradation product and the reason for the degradation of product II were investigated using HPLC. One biotransformation product, daphnetin-8-O-beta-D-glucoside, was obtained, the optimal co-cultured time in suspension hairy root of P. multiflorum was 36 h with the highest biotransformation molar ratio of 32.11%, the sucrose medium (sucrose-only) can increase the biotransformation molar ratio to 72.44%. The result demonstrated that the degradation products of the product II was induced by the MS medium. The potential application of suspension transgenic hairy root of P. multflorum in the sucrose-only medium on generating daphnetin-8-Obeta-D- glucoside could be prospective.

  17. Involvement of ethylene and lipid signalling in cadmium-induced programmed cell death in tomato suspension cells

    NARCIS (Netherlands)

    Yakimova, E.T.; Kapchina-Toteva, V.M.; Laarhoven, L.J.J.; Harren, F.J.M.; Woltering, E.J.

    2006-01-01

    Cadmium-induced cell death was studied in suspension-cultured tomato (Lycopersicon esculentum Mill.) cells (line MsK8) treated with CdSO4. Within 24 h, cadmium treatment induced cell death in a concentration-dependent manner. Cell cultures showed recovery after 23 days which indicates the existence

  18. Involvement of ethylene and lipid signalling in cadmium-induced programmed cell death in tomato suspension cells

    NARCIS (Netherlands)

    Iakimova, E.T.; Kapchina-Toteva, V.M.; Laarhoven, L.J.; Harren, F.; Woltering, E.J.

    2006-01-01

    Cadmium-induced cell death was studied in suspension-cultured tomato (Lycopersicon esculentum Mill.) cells (line MsK8) treated with CdSO4. Within 24 h, cadmium treatment induced cell death in a concentration-dependent manner. Cell cultures showed recovery after 2¿3 days which indicates the existence

  19. Clogging in constricted suspension flows

    Science.gov (United States)

    Marin, Alvaro; Lhuissier, Henri; Rossi, Massimiliano; Kähler, Christian J.

    2018-02-01

    The flow of a charged-stabilized suspension through a single constricted channel is studied experimentally by tracking the particles individually. Surprisingly, the behavior is found to be qualitatively similar to that of inertial dry granular systems: For small values of the neck-to-particle size ratio (D /d reported for granular systems and agree for moderate particle volume fraction (ϕ ≈20 % ) with a simple stochastic model for the number of particles at the neck. For larger neck sizes (D /d >3 ), even at the largest ϕ (≈60 %) achievable in the experiments, an uninterrupted particle flow is observed, which resembles that of an hourglass. This particularly small value of D /d (≃3 ) at the transition to a practically uninterrupted flow is attributed to the low effective friction between the particles, achieved by the particle's functionalization and lubrication.

  20. Next Generation Suspension Dynamics Algorithms

    Energy Technology Data Exchange (ETDEWEB)

    Schunk, Peter Randall [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Higdon, Jonathon [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Chen, Steven [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2014-12-01

    This research project has the objective to extend the range of application, improve the efficiency and conduct simulations with the Fast Lubrication Dynamics (FLD) algorithm for concentrated particle suspensions in a Newtonian fluid solvent. The research involves a combination of mathematical development, new computational algorithms, and application to processing flows of relevance in materials processing. The mathematical developments clarify the underlying theory, facilitate verification against classic monographs in the field and provide the framework for a novel parallel implementation optimized for an OpenMP shared memory environment. The project considered application to consolidation flows of major interest in high throughput materials processing and identified hitherto unforeseen challenges in the use of FLD in these applications. Extensions to the algorithm have been developed to improve its accuracy in these applications.

  1. Naratriptan hydrochloride in extemporaneosly compounded oral suspensions.

    Science.gov (United States)

    Zhang, Y P; Trissel, L A; Fox, J L

    2000-01-01

    The purpose of this study was to determine the pharmaceutical acceptability and chemical stability of naratriptan hydrochloride in three extemporaneously compounded suspension formulations. The naratriptan-hydrochloride oral suspensions were prepared from 2.5-mg commercial tablets yielding a nominal naratriptan concentration of 0.5 mg/mL. The suspension vehicles selected for testing were Syrpalta, an equal-parts mixture of Ora-Plus and Ora-Sweet, and an equal-parts mixture of Ora-Plus and Ora-Sweet SF. The tablets were crushed and thoroughly triturated to a fine powder using a porcelain mortar and pestle. The powder was incorporated into a portion of the Syrpalta or Ora-Plus suspension vehicle and mixed until homogeneous. The mixtures were then brought to volume with Syrpalta, Ora-Sweet or Ora-Sweet SF, as appropriate. The suspensions were packaged in amber, plastic, screw-cap prescription bottles and stored at 23 deg C for seven days and 4 deg C for 90 days. An adequate suspension was never achieved in Syrpalta. The crushed-tablet powder did not produce a uniformly dispersed mixture and exhibited clumping and a high rate of sedimentation. A distinct layer of the solid tablet material settled immediately after shaking. Over the next four hours, a densely packed, yellow, caked layer formed at the bottom of the containers, making resuspension difficult. During storage, the caking became worse. Chemical analysis was not performed. The Ora-Plus and Ora-Sweet or Ora-Sweet SF suspensions had a slight greenish cast and were resuspended without difficulty by shaking for approximately ten seconds, yielding easily poured and homogeneous mixtures throughout the study. Visible settling and layering did not begin for four hours with the Ora-Sweet suspension and 24 hours for the Ora-Sweet SF suspension. High pressure liquid chromatographic analysis found that the naratriptan concentration in both suspension-vehicle combinations exhibited little or no loss for seven days at 23

  2. Optimization of callus and cell suspension cultures of Barringtonia racemosa (Lecythidaceae family for lycopene production Otimização de culturas de suspensões de calos e células de Barringtonia racemosa (família Lecythidaceae para produção de licopeno

    Directory of Open Access Journals (Sweden)

    Mandana Behbahani

    2011-02-01

    Full Text Available Lycopene is present in a range of fresh fruits and vegetables, especially in the leaves of Barringtonia racemosa. The traditional lycopene extraction from the plant is being employed instead of an easy propagation technique like cell culture process from the leaf explants. We intend to assess how lycopene could be extracted via tissue culture under light (illuminance: 8,200 lux under white fluorescent lamps, photoperiod 16 h per day at 25ºC and dark. Leaf explants of Barringtonia racemosa were cultured on modified Murashige and Skoog (MS, Woody Plant Medium (WPM and B5 media, supplemented with different concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D. Optimal conditions for callus induction and maintenance under both dark and light were investigated, and growth and lycopene accumulation were evaluated. Among media with different concentrations of 2,4-D, fast growing, friable callus initiated within three weeks after culturing on WPM basal medium supplemented with 2.0 mg L-1 (weight per volume of 2,4-D, whereas callus induction in explants cultured on all other media started only after five weeks. Calli were subcultured once every fortnight. Pale yellow and green calli developed under conditions of dark and light respectively were then selected for evaluation of their lycopene contents. An improved reversed phase of high performance liquid chromatography (HPLC method was used for a selective chemical determination of the lycopene content. Light induced lycopene production; and likewise maximum lycopene level incubated in light was higher than those incubated in darkness. The best growth rates of callus and cell suspension were achieved in WPM and B5 media respectively. The production of lycopene was growth-dependent through analysis of growth and lycopene content of both callus and cell suspension cultures.O licopeno está presente numa série de frutas frescas e hortaliças principalmente na folhas de Barringtonia racemosa. A extra

  3. Monitoring of the region’s socio-cultural development as a component to enhance the effectiveness of social administration

    Directory of Open Access Journals (Sweden)

    Mariya Aleksandrovna Lastochkina

    2014-11-01

    Full Text Available The article presents the results of the next phase of the study on the subject “Socio-cultural portrait of the region”. It gives the estimation of the demographic situation, standard of living and welfare of the Vologda Oblast residents. Besides, the article shows the people’s assessment of the performance of the authorities, social well-being and degree of safety, as well as their trust in the government. All these aspects are studied in their connection with social policy and social protection measures undertaken in the region. The study reveals that the society is disappointed in the actions of the authorities; it does not understand the true ultimate effect and the feasibility of ongoing regional social events. The greatest number of critical remarks addressed to local authorities comes from people with unstable position in life. Negative evaluations are also more likely expressed by the poorest population groups, by women, by the less educated, by village dwellers and urban-type settlements residents, as well as by individuals with a low level of education. Social policy in t he minds of a large part of the population is associated with principles of social justice, equality, rule of law, responsibility and socially important values. The Vologda Oblast residents do not feel that their spiritual and ideological views are being endangered; and the issues associated with ensuring the security of life and health, decent standard of living, legitimate and fair performance of the basic social institutions remain unsettled, according to their understanding. At the same time, social policy should create conditions to stabilize social relations, to ensure efficiency and sustainability in the development of the whole society, which should adequately respond to all positive and negative effects

  4. Bridging cross-cultural gaps: monitoring herbal use during chemotherapy in patients referred to integrative medicine consultation in Israel.

    Science.gov (United States)

    Almog, Limor; Lev, Efraim; Schiff, Elad; Linn, Shai; Ben-Arye, Eran

    2014-10-01

    The high prevalence of the use of traditional herbs among patients with cancer is a cause for concern with regard to potentially adverse interactions with conventional oncology treatments. In this study, we explore herbal use among patients with cancer in northern Israel who are referred by their health care providers to complementary and traditional medicine (CTM) consultations provided to them within the conventional oncology department. The study's objectives were to identify which herbs patients use and to examine the scope of current research on the efficacy and safety regarding the identified herbs. Herbal use by patients receiving oncology care was assessed prospectively from July 2009 to July 2012 by integrative physicians (IPs) trained in herbal medicine. Historical, ethnobotanical, basic research, and clinical data regarding the identified herbs were explored by using a keyword search in PubMed and Middle Eastern ethnohistorical literature. Disclosure of herbal use was reported by 154 of the 305 patients (50.5 %) interviewed by IPs. The use of 85 single herbs and 30 different herbal formulas was documented during the initial or follow-up IP assessments. Patients reported 14 quality of life-associated indications for herbal use. The ten most prevalent herbs displaying in vitro/in vivo anticancer activity and nine other herbs were preliminarily assessed concerning potential risks, safety, and interaction with chemotherapy. Herbal use by patients with cancer in northern Israel is widespread and calls for further study in order to address issues of safety and effectiveness. We recommend constructing a multinational and multidisciplinary team of researchers with ethnopharmacological and clinical expertise that will explore the use of herbs among patients with cancer in a cross-cultural perspective attuned with patients' affinity to traditional herbal medicine.

  5. Safety culture

    International Nuclear Information System (INIS)

    Drukraroff, C.

    2010-01-01

    The concept of Safety Culture was defined after Chernobyl's nuclear accident in 1986. It has not been exempt from discussion interpretations, adding riders, etc..., over the last 24 years because it has to do with human behavior and performance in the organizations. Safety Culture is not an easy task to define, assess and monitor. The proof of it is that today we still discussing and writing about it. How has been the evolution of Safety Culture at the Juzbado Factory since 1985 to today?. What is the strategy that we will be following in the future. (Author)

  6. Fifth International Symposium on Magnetic Suspension Technology

    Science.gov (United States)

    Groom, Nelson J. (Editor); Britcher, Colin P.

    2000-01-01

    In order to examine the state of technology of all areas of magnetic suspension and to review recent developments in sensors, controls, superconducting magnet technology, and design/implementation practices, the Fifth International Symposium on Magnetic Suspension Technology was held at the Radisson Hotel Santa Barbara, Santa Barbara, California, on December 1-3, 1999. The symposium included 18 sessions in which a total of 53 papers were presented. The technical sessions covered the areas of bearings, controls, modeling, electromagnetic launch, magnetic suspension in wind tunnels, applications flywheel energy storage, rotating machinery, vibration isolation, and maglev. A list of attendees is included in the document.

  7. Vehicle lateral dynamics stabilization using active suspension

    Directory of Open Access Journals (Sweden)

    Drobný V.

    2008-12-01

    Full Text Available The paper deals with the investigation of active nonlinear suspension control in order to stabilize the lateral vehicle motion in similar way as systems like ESP do. The lateral stabilization of vehicle based on braking forces can be alternatively provided by the different setting of suspension forces. The basis of this control is the nonlinear property of the tyres. The vehicle has at least four wheels and it gives one or more redundant vertical forces that can be used for the different distribution of vertical suspension forces in such a way that resulting lateral and/or longitudinal forces create the required correction moment for lateral dynamic vehicle stabilization.

  8. Suspension for the low frequency facility

    CERN Document Server

    Cella, G; Di Virgilio, A; Gaddi, A; Viceré, A

    2000-01-01

    We introduce the working principles of the VIRGO Low Frequency Facility (LFF), whose main aim is the measurement of the thermal noise in the VIRGO suspension system. We evaluate the displacement thermal noise of a mirror, which is an intermediate element of a double pendulum suspension system. This double pendulum will be suspended to the last stage of a VIRGO Super-Attenuator (SA), the prototype VIRGO suspension system being tested at the Pisa section of INFN. In the proposed configuration, we evaluate the spectrum of the thermal noise for different choices of the parameters: based on this study, we comment on the future directions to be undertaken in the LFF experiment.

  9. Conditions for a high plating efficiency of free cell suspensions of Haplopappus gracilis (Nutt) gray.

    Science.gov (United States)

    Werry, P A; Stoffelsen, K M

    1978-07-01

    Suspensions of Haplopappus gracilis cells, containing about 80% free cells, were obt ained from log-phase cultures by filtration through 3 nylon sieves having decreasing mesh widths from 297, 210 and 88 μm. From the free cell suspensions, 75 to 90% of the cells developed into visible colonies when the plating procedure was divided into two steps: a) plating the cells at high concentration in soft agar on feeder agar; b) replating the resulting aggregations at appropriate concentrations on fresh feeder agar. From the results, it is inferred that, in the replating step, the volume of the inoculum is the deciding factor which influences the resulting plating efficiency.

  10. Gravisensing, apoptosis, and drug recovery in Taxus cell suspensions

    Science.gov (United States)

    Durzan, D. J.

    1999-01-01

    Haploid and diploid cell suspensions of Taxus spp. were examined for their adaptive plasticity in response to simulated microgravity, unit gravity, and hypergravity. Cell suspensions produced the taxane, paclitaxel, (TAXOL (R)), which is useful for the treatment of various cancers. Amyloplasts contributed to taxane ring biosynthesis and to drug release at the cell wall. Drug-producing cells reacted as gravisensing osmotic tensiometers. In stressed cells, amyloplasts docked and fused in clusters to sites on the plasmalemma before taxane discharge into the culture medium. In simulated microgravity and compared to all other treatments, taxane production was reduced nearly 100-fold. The percent paclitaxel of total taxanes remained 3-to 6-fold greater, and biomass doubled. When p53-independent programmed cell death was induced, taxanes were released into the culture medium as free molecules (soluble and insoluble) or bound to membranes, nuclear fragments, xylan residues, and other particulate materials. Unit gravity and especially hypergravity promoted xylogenesis and significant drug overproduction. A model relating families of >touch = (TCH), taxane early response (TER), nuclear cycling, and apoptosis-regulating genes to gravisensing, cell wall modifications, and to taxane recovery accounted for most but not all of the observations.

  11. 48 CFR 42.1302 - Suspension of work.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 1 2010-10-01 2010-10-01 false Suspension of work. 42... MANAGEMENT CONTRACT ADMINISTRATION AND AUDIT SERVICES Suspension of Work, Stop-Work Orders, and Government Delay of Work 42.1302 Suspension of work. A suspension of work under a construction or architect...

  12. 12 CFR 367.8 - Causes for suspension.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 4 2010-01-01 2010-01-01 false Causes for suspension. 367.8 Section 367.8... SUSPENSION AND EXCLUSION OF CONTRACTOR AND TERMINATION OF CONTRACTS § 367.8 Causes for suspension. (a... suspension by another federal agency; (2) That a cause for exclusion under § 367.6 may exist; (3) Of the...

  13. 45 CFR 1641.13 - Causes for suspension.

    Science.gov (United States)

    2010-10-01

    ... 45 Public Welfare 4 2010-10-01 2010-10-01 false Causes for suspension. 1641.13 Section 1641.13..., SUSPENSION AND REMOVAL OF RECIPIENT AUDITORS Suspension § 1641.13 Causes for suspension. The debarring... that: (a) A cause for debarment under § 1641.7 may exist; (b) The IPA has been indicted for or...

  14. callus culture

    African Journals Online (AJOL)

    steve

    capacity in comparison to those of grown on 75%. NH4NO3 depleted media under the same light regime. Light dependent secondary metabolite production. Initially, we have cultured wounded grape leaf explants in the dark for callus induction and monitoring possible pigmentation and phenolics accumulation. Supplemen-.

  15. Stimulation of taxane production in suspension cultures of Taxus ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-06-17

    Jun 17, 2008 ... To improve the paclitaxel yield various kinds of elicitors such as methyl jasmonate, fungal elicitor, chitosan and silver ion have been applied and very effective for ... 81325, Fluka) by the procedure of Roco et al. (1993) with modification. Polygalac- turonic acid at 50 µg ml-1 with pectinase (Cat. 17389, Fluka) ...

  16. In vitro plant regeneration from embryogenic cell suspension culture ...

    African Journals Online (AJOL)

    After two weeks, induction of somatic embryos up to the torpedo stage occured at all tested concentrations of 2,4-D, IAA or NAA. Somatic embryos developed only in MS medium containing 0.5 mg/l IAA within two weeks and 2% of globular embryos were developed into the cotyledonary stage embryos. Eighty one percent of ...

  17. Regeneration from embryogenic callus and suspension cultures of ...

    African Journals Online (AJOL)

    ehab

    2012-04-25

    Apr 25, 2012 ... source of palmerosa, citronella, lemon-grass and oil of. Malabar, respectively. C. jwarancusa is used as condiment and for medicinal purpose. ... and rinsed under running tap water for 15 min, then washed with. El-Bakry and Abdel-Salam 10099 double distilled H2O (ddH2O). Seeds were then treated in 95 ...

  18. Stimulation of taxane production in suspension cultures of Taxus ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-06-17

    Jun 17, 2008 ... Planta 215: 596-605. Nothnagel EA, McNeil M, Albersheim P, Dell A (1983). Host pathogen interaction XXII: a galacturonic acid oligosaccharide from plant cell wall elicits phytoalexins. Plant Physiol. 71: 916-926. Ramesh NP (1998). Tour de paclitaxel: biocatalysis for semisynthesis. Annu. Rev. Microbiol.

  19. Inert Cities: Globalization, Mobility and Suspension in Visual Culture

    NARCIS (Netherlands)

    Donald, S.H.; Lindner, C.

    2014-01-01

    We usually associate contemporary urban life with movement and speed. But what about those instances when the forms of mobility associated with globalized cities - the flow of capital, people, labor and information - freeze, or decelerate? How can we assess the value of interruption in a city? What

  20. Satellite SAR imagery for site discovery, change detection and monitoring activities in cultural heritage sites: experiments on the Nasca region, Peru

    Science.gov (United States)

    Tapete, D.; Cigna, F.; Masini, N.; Lasaponara, R.

    2012-04-01

    data stacks were selected: ERS-2 ascending data acquired in 2001-2011, ENVISAT ASAR ascending and descending data acquired in 2003-2007, and ALOS PALSAR descending and ascending data acquired in 2007 and 2008. The feature extraction was specifically addressed to the recognition of buried structures, archaeological deposits and the study of the buried networks of aqueducts, as well as the morphological study of the Nasca geoglyphs. Change detection analysis also included the multi-temporal reconstruction of the evolution of the Rio Nasca catchment basin, while specific tests were performed to demonstrate the usefulness of SAR imagery for monitoring looting activities. The results of the radar-interpretation compared and integrated with the field investigations will support the archaeological activities and contribute to the monitoring and enhancement of archaeological heritage and cultural landscape of the Nasca region.

  1. A Course in Fluid Mechanics of Suspensions.

    Science.gov (United States)

    Davis, Robert H.

    1989-01-01

    Discusses a course focusing on fluid mechanics and physical chemistry of suspensions. Describes the main themes of the lectures and includes a list of course outlines. Possible textbooks and many journal articles are listed. (YP)

  2. Experimental Evaluation of Mountain Bike Suspension Systems

    Directory of Open Access Journals (Sweden)

    J. Titlestad

    2003-01-01

    Full Text Available A significant distinction between competitive mountain bikes is whether they have a suspension system. Research studies indicate that a suspension system gives advantages, but it is difficult to quantify the benefits because they depend on so many variables, including the physiology and psychology of the cyclist, the roughness of the track and the design of the suspension system. A laboratory based test rig has been built that allows the number of variables in the system to be reduced and test conditions to be controlled. The test rig simulates regular impacts of the rear wheel with bumps in a rolling road. The physiological variables of oxygen consumption and heart rate were measured, together with speeds and forces at various points in the system. Physiological and mechanical test results both confirm a significant benefit in using a suspension system on the simulated rough track, with oxygen consumption reduced by around 30 % and power transmitted through the pedals reduced by 30 % to 60 %.

  3. 78 FR 17133 - Suspension of Community Eligibility

    Science.gov (United States)

    2013-03-20

    ... scheduled for suspension on the effective dates listed within this rule because of noncompliance with the... participation status of a community can be obtained from FEMA's Community Status Book (CSB). The CSB is... [[Page 17134

  4. Heat and mass transfer in particulate suspensions

    CERN Document Server

    Michaelides, Efstathios E (Stathis)

    2013-01-01

    Heat and Mass Transfer in Particulate Suspensions is a critical review of the subject of heat and mass transfer related to particulate Suspensions, which include both fluid-particles and fluid-droplet Suspensions. Fundamentals, recent advances and industrial applications are examined. The subject of particulate heat and mass transfer is currently driven by two significant applications: energy transformations –primarily combustion – and heat transfer equipment. The first includes particle and droplet combustion processes in engineering Suspensions as diverse as the Fluidized Bed Reactors (FBR’s) and Internal Combustion Engines (ICE’s). On the heat transfer side, cooling with nanofluids, which include nanoparticles, has attracted a great deal of attention in the last decade both from the fundamental and the applied side and has produced several scientific publications. A monograph that combines the fundamentals of heat transfer with particulates as well as the modern applications of the subject would be...

  5. Modeling tensorial conductivity of particle suspension networks.

    Science.gov (United States)

    Olsen, Tyler; Kamrin, Ken

    2015-05-21

    Significant microstructural anisotropy is known to develop during shearing flow of attractive particle suspensions. These suspensions, and their capacity to form conductive networks, play a key role in flow-battery technology, among other applications. Herein, we present and test an analytical model for the tensorial conductivity of attractive particle suspensions. The model utilizes the mean fabric of the network to characterize the structure, and the relationship to the conductivity is inspired by a lattice argument. We test the accuracy of our model against a large number of computer-generated suspension networks, based on multiple in-house generation protocols, giving rise to particle networks that emulate the physical system. The model is shown to adequately capture the tensorial conductivity, both in terms of its invariants and its mean directionality.

  6. Suspension scheme for fuel pin

    International Nuclear Information System (INIS)

    Butts, C.E.; Gray, H.C.

    1975-01-01

    A description is presented of a nuclear fuel pin suspension arrangement comprising, in combination, a rod; a first beam member connected to said rod at one end; a plurality of parallel-spaced slidable fuel support plates attached to said first beam member, the longitudinal axis of first beam member being perpendicular to the longitudinal axis of each of said fuel support plates, a first coupling means disposed along the length of the first beam member for permitting slidable fuel support plates parallel movement with respect to the longitudinal axis of said first beam member, a second coupling means located at one end of each of slidable fuel plates for slidably engaging first coupling means of first beam member, a second beam member connected to the other end of each of parallel-spaced slidable fuel support plates and providing an extension, second beam member being provided with a third coupling means disposed along the length of second beam member at one end thereof; and a plurality of fuel pins provided with a fourth coupling means located at one end of each fuel pin for slidably engaging third coupling means of second beam member to permit each fuel pin parallel movement with respect to the longitudinal axis of second beam member. (U.S.)

  7. Increasing Possibilities of Nano suspension

    International Nuclear Information System (INIS)

    Sutradhar, K.B.; Khatun, S.; Luna, I.P.

    2013-01-01

    Nowadays, a very large proportion of new drug candidates emerging from drug discovery programmes are water insoluble and thus poorly bioavailable. To avoid this problem, nano technology for drug delivery has gained much interest as a way to improve the solubility problems. Nano refers to particles size range of 1-1000 nm. The reduction of drug particles into the submicron range leads to a significant increase in the dissolution rate and therefore enhances bioavailability. Nanosuspensions are part of nano technology. This interacts with the body at subcellular (i.e., molecular) scales with a high degree of specificity and can be potentially translated into targeted cellular and tissue-specific clinical applications designed to achieve maximal therapeutic efficacy with minimal side effects. Production of drugs as nanosuspensions can be developed for drug delivery systems as an oral formulation and no noral administration. Here, this review describes the methods of pharmaceutical nano suspension production including advantages and disadvantages, potential benefits, characterization tests, and pharmaceutical applications in drug delivery

  8. Characterization of Complex Colloidal Suspensions

    Science.gov (United States)

    Seaman, J. C.; Guerin, M.; Jackson, B. P.; Ranville, J. M.

    2003-04-01

    Surface chemical reactions play a major role in controlling contaminant fate and transport in the subsurface environment. Recent field and laboratory evidence suggests that mobile soil and groundwater colloids may facilitate the migration of sparingly soluble groundwater contaminants. Colloidal suspensions collected in the field or generated in laboratory column experiments tend to be fairly dilute in nature and comprised of relatively small particulates (reserved for studying ideal systems to the characterization of mobile colloids. However, many of these analytical techniques, including total/selective dissolution methods, dynamic light scattering, micro-electrophoresis, streaming potential, and even scanning electron microscopy (SEM), can be biased in of larger size fractions, and therefore, extremely sensitive to sampling, storage, and fractionation artifacts. In addition, surface modifiers such as sorbed oxides or organics can alter particulate appearance, composition, and behavior when compared to synthetic analogues or mineral standards. The current presentation will discuss the limitations and inherent biases associated with a number of analytical characterization techniques that are commonly applied to the study of mobile soil and groundwater colloids, including field flow fractionation (FFF) and acoustic based methods that have only recently become available.

  9. Core Muscle Activation in Suspension Training Exercises

    OpenAIRE

    Cugliari, Giovanni; Boccia, Gennaro

    2017-01-01

    Abstract A quantitative observational laboratory study was conducted to characterize and classify core training exercises executed in a suspension modality on the base of muscle activation. In a prospective single-group repeated measures design, seventeen active male participants performed four suspension exercises typically associated with core training (roll-out, bodysaw, pike and knee-tuck). Surface electromyographic signals were recorded from lower and upper parts of rectus abdominis, ext...

  10. Compressed Air Production Using Vehicle Suspension

    OpenAIRE

    Ninad Arun Malpure; Sanket Nandlal Bhansali

    2015-01-01

    Abstract Generally compressed air is produced using different types of air compressors which consumes lot of electric energy and is noisy. In this paper an innovative idea is put forth for production of compressed air using movement of vehicle suspension which normal is wasted. The conversion of the force energy into the compressed air is carried out by the mechanism which consists of the vehicle suspension system hydraulic cylinder Non-return valve air compressor and air receiver. We are co...

  11. Monitoring liquid and solid content in froth using conductivity

    Science.gov (United States)

    J.Y. Zhu; F. Tan; R. Gleisner

    2005-01-01

    This study reports the feasibility of monitoring liquid and fiber rejection during froth flotation of fiber suspensions through conductivity measurements of the rejected froth. The technique was demonstrated in laboratory flotation experiments using nylon and wood fiber suspensions in two laboratory flotation cells. We found that both the total wet rejection and the...

  12. Core muscle activity during suspension exercises.

    Science.gov (United States)

    Mok, Nicola W; Yeung, Ella W; Cho, Jeran C; Hui, Samson C; Liu, Kimee C; Pang, Coleman H

    2015-03-01

    Suspension exercise has been advocated as an effective means to improve core stability among healthy individuals and those with musculoskeletal complaints. However, the activity of core muscles during suspension exercises has not been reported. In this study, we investigated the level of activation of core muscles during suspension exercises within young and healthy adults. The study was conducted in a controlled laboratory setting. Surface electromyographic (sEMG) activity of core muscles (rectus abdominis, external oblique, internal oblique/transversus abdominis, and superficial lumbar multifidus) during four suspension workouts (hip abduction in plank, hamstring curl, chest press, and 45° row) was investigated. Muscle activity during a 5-s hold period of the workouts was measured by sEMG and normalized to the individual's maximal voluntary isometric contraction (MVIC). Different levels of muscle activation were observed during the hip abduction in plank, hamstring curl, and chest press. Hip abduction in plank generated the highest activation of most abdominal muscles. The 45° row exercise generated the lowest muscle activation. Among the four workouts investigated, the hip abduction in plank with suspension was found to have the strongest potential strengthening effect on core muscles. Also, suspension training was found to generate relatively high levels of core muscle activation when compared with that among previous studies of core exercises on stable and unstable support surfaces. Copyright © 2014 Sports Medicine Australia. Published by Elsevier Ltd. All rights reserved.

  13. Extensional viscosity of microfibrillated cellulose suspensions.

    Science.gov (United States)

    Moberg, Tobias; Rigdahl, Mikael; Stading, Mats; Levenstam Bragd, Emma

    2014-02-15

    The extensional properties of micro fibrillated cellulose (MFC)-suspensions at different fibril concentrations and with different amounts of added sodium chloride were evaluated. The MFC-suspensions were obtained by diluting a stock solution consisting of 0.95 wt.% cellulose with either deionized water or sodium chloride solution, giving a series of different concentrations and sodium chloride contents. The extensional viscosities of the suspensions were measured utilizing contraction flow geometry. Here the specimens were forced through a hyperbolic nozzle and the required pressure drop over the nozzle was measured. The extensional viscosity exhibited an extensional-thinning behaviour over the extensional strain rates used. Furthermore the extensional viscosity decreased with decreasing concentration of the suspensions, in similarities with the shear properties of the specimens. For the suspensions containing sodium chloride, the extensional viscosity appeared to increase when the concentration of sodium chloride was increased. But excessive amounts of added sodium chloride promoted an agglomeration of the suspensions. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Economic impact of cultural tourism

    OpenAIRE

    Zadel, Zrinka; Bogdan, Sinisa

    2013-01-01

    The subject of analysis in the paper is economic impact of cultural tourism and identification of the main factors which directly affect cultural tourism revenues. Most countries do not have a statistical system of monitoring and analysing individual factors of cultural tourism such as the number of arrivals of cultural tourists and consumption of cultural tourists. Therefore, it is hard to assess the economic impact of cultural tourism. In cultural tourism, cultural assets are prepared and p...

  15. Monitoring of Legionella pneumophila viability after chlorine dioxide treatment using flow cytometry.

    Science.gov (United States)

    Mustapha, Pascale; Epalle, Thibaut; Allegra, Séverine; Girardot, Françoise; Garraud, Olivier; Riffard, Serge

    2015-04-01

    The viability of three Legionella pneumophila strains was monitored after chlorine dioxide (ClO2) treatment using a flow cytometric assay. Suspensions of L. pneumophila cells were submitted to increasing concentrations of ClO2. Culturable cells were still detected when using 4 mg/L, but could no longer be detected after exposure to 6 mg/L of ClO2, although viable but not culturable (VBNC) cells were found after exposure to 4-5 mg/L of ClO2. When testing whether these VBNC were infective, two of the strains were resuscitated after co-culture with Acanthamoeba polyphaga, but neither of them could infect macrophage-like cells. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  16. Complementary analysis techniques applied on optimizing suspensions of yttria stabilized zirconia

    DEFF Research Database (Denmark)

    Della Negra, Michela; Foghmoes, Søren Preben Vagn; Klemensø, Trine

    2016-01-01

    Three different polymers with different functional groups and similar molecular weight were tested as dispersing agents for suspensions of yttria stabilized zirconia in ethanol: polyvinyl pyrrolidone, polyethylene imine, polyvinyl butyral/acetal. The stability of the system was assessed consideri...... excellent performance of polyvinyl pyrrolidone and polyethylene imine as dispersing agents. The stability and dispersing power were finally utilized for preparing concentrated suspensions for tape casting and subsequently to sinter the tapes into dense ceramic pieces......., in details, all the processing steps, including suspension de-agglomeration, slurry manipulation, quality of sintered tapes microstructure, and final layer leak tightness. Different analytical techniques were used to monitor ceramic de-agglomeration and stability as a function of time, for different types...

  17. Intercomparison of Numerical Inversion Algorithms for Particle Size Determination of Polystyrene Suspensions Using Spectral Turbidimetry

    Directory of Open Access Journals (Sweden)

    Benjamin Glasse

    2015-01-01

    Full Text Available The continuous monitoring of the particle size distribution in particulate processes with suspensions or emulsions requires measurement techniques that can be used as in situ devices in contrast to ex situ or laboratory methods. In this context, for the evaluation of turbidimetric spectral measurements, the application of different numerical inversion algorithms is investigated with respect to the particle size distribution determination of polystyrene suspensions. A modified regularization concept consisting of a Twomey-Phillips-Regularization with an integrated nonnegative constraint and a modified L-curve criterion for the selection of the regularization parameter is used. The particle size (i.e., particle diameter of polystyrene suspensions in the range x=0.03–3 µm was validated via dynamic light scattering and differential centrifugal sedimentation and compared to the retrieved particle size distribution from the inverted turbidimetry measurements.

  18. Culture conditions have an impact on the maturation of traceable, transplantable mouse embryonic stem cell-derived otic progenitor cells.

    Science.gov (United States)

    Abboud, Nesrine; Fontbonne, Arnaud; Watabe, Isabelle; Tonetto, Alain; Brezun, Jean Michel; Feron, François; Zine, Azel

    2017-09-01

    The generation of replacement inner ear hair cells (HCs) remains a challenge and stem cell therapy holds the potential for developing therapeutic solutions to hearing and balance disorders. Recent developments have made significant strides in producing mouse otic progenitors using cell culture techniques to initiate HC differentiation. However, no consensus has been reached as to efficiency and therefore current methods remain unsatisfactory. In order to address these issues, we compare the generation of otic and HC progenitors from embryonic stem (ES) cells in two cell culture systems: suspension vs. adherent conditions. In the present study, an ES cell line derived from an Atoh1-green fluorescent protein (GFP) transgenic mouse was used to track the generation of otic progenitors, initial HCs and to compare these two differentiation systems. We used a two-step short-term differentiation method involving an induction period of 5 days during which ES cells were cultured in the presence of Wnt/transforming growth factor TGF-β inhibitors and insulin-like growth factor IGF-1 to suppress mesoderm and reinforce presumptive ectoderm and otic lineages. The generated embryoid bodies were then differentiated in medium containing basic fibroblast growth factor (bFGF) for an additional 5 days using either suspension or adherent culture methods. Upon completion of differentiation, quantitative polymerase chain reaction analysis and immunostaining monitored the expression of otic/HC progenitor lineage markers. The results indicate that cells differentiated in suspension cultures produced cells expressing otic progenitor/HC markers at a higher efficiency compared with the production of these cell types within adherent cultures. Furthermore, we demonstrated that a fraction of these cells can incorporate into ototoxin-injured mouse postnatal cochlea explants and express MYO7A after transplantation. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons

  19. Suspension, a Wake-Up Call: Rural Educators' Attitudes toward Suspension.

    Science.gov (United States)

    Henderson, Joan; Friedland, Billie

    Data from the West Virginia Department of Education reveals that from September 1991 to January 1992, school districts reported 18,915 out-of-school suspensions involving 12,997 students. In 1995, the West Virginia State Legislature enacted the Safe Schools Act, which specifically mandates suspension for no less than 12 consecutive months for…

  20. On-line monitoring of important organoleptic methyl-branched aldehydes during batch fermentation of starter culture Staphylococcus xylosus reveal new insight into their production in a model fermentation

    DEFF Research Database (Denmark)

    de Vos Petersen, Christian; Beck, Hans Christian; Lauritsen, Frants R

    2004-01-01

    of the very important flavor compounds 2-methylbutanal, 3-methylbutanal, and 2-methylpropanal with various growth conditions. We found that the aldehydes were present in the culture broth only as transient metabolites. They were produced in the exponential growth phase, reached a maximum concentration when......A small fermentor (55 mL) was directly interfaced to a membrane inlet mass spectrometer for continuous on-line monitoring of oxygen and volatile metabolites during batch fermentations of the starter culture Staphylococcus xylosus. Using this technique, we were able to correlate production...... the culture became anaerobic, and then they rapidly disappeared from the culture medium. This general pattern was observed for three different strains of S. xylosus and S. carnosus. Small amounts of inoculum or increased exposure to oxygen were found to favor production of the aldehydes as a result...

  1. Rheological properties of nanocrystalline cellulose suspensions.

    Science.gov (United States)

    Chen, Yang; Xu, Chunjiang; Huang, Jing; Wu, Defeng; Lv, Qiaolian

    2017-02-10

    Rheological behavior, including linear and nonlinear, as well as transient rheology of nanocrystalline cellulose (NCC) suspensions was studied in this work. Two kinds of polymer solutions, aqueous poly(vinyl alcohol) (PVA) with flexible chain structure and aqueous carboxymethyl cellulose (CMC) with semi-rigid chain structure, were used as the suspension media to further explore the role that the interactions among NCC and polymers played during shear flow. The results reveal that NCC has lower values of percolation threshold in the PVA solution than in the CMC one during small amplitude oscillatory shear (SAOS) flow because the flexible PVA chain has higher adsorbed level onto NCC particles than the negatively charged semi-rigid CMC chain, which is further confirmed by the Fourier transformed infrared (FT-IR) spectroscopy tests. As a result, the NCC suspension shows a weak strain overshoot in PVA solution during large amplitude oscillatory shear (LAOS) flow, which cannot be seen on the one in CMC solution. During startup shear flow, both of these two suspensions show evident stress overshoot behavior with the strain-scaling characteristics, indicating the formation of ordered long-term structure of rod-like NCC particles with self-similarity during flow. However, NCC suspension have far stronger stress overshoot response in CMC solution relative to the one in PVA solution. A possible synergy mechanism between NCC and CMC chain is hence proposed. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Numerical study of suspensions of deformable particles.

    Science.gov (United States)

    Brandt, Luca; Rosti, Marco Edoardo

    2017-11-01

    We consider a model non-Newtonian fluid consisting of a suspension of deformable particles in a Newtonian solvent. Einstein showed in his pioneering work that the relative increase in effective viscosity is a linear function of the particle volume fraction for dilute suspensions of rigid particles. Inertia has been shown to introduce deviations from the behaviour predicted by the different empirical fits, an effect that can be related to an increase of the effective volume fraction. We here focus on the effect of elasticity, i.e. visco-elastic deformable particles. To tackle the problem at hand, we perform three-dimensional Direct Numerical Simulation of a plane Couette flow with a suspension of neutrally buoyant deformable viscous hyper-elastic particles. We show that elasticity produces a shear-thinning effect in elastic suspensions (in comparison to rigid ones) and that it can be understood in terms of a reduction of the effective volume fraction of the suspension. The deformation modifies the particle motion reducing the level of mutual interaction. Normal stress differences will also be considered. European Research Council, Grant No. ERC-2013-CoG- 616186, TRITOS; SNIC (the Swedish National Infrastructure for Computing).

  3. A review of dynamics modelling of friction wedge suspensions

    Science.gov (United States)

    Wu, Qing; Cole, Colin; Spiryagin, Maksym; Sun, Yan Quan

    2014-11-01

    Three-piece bogies with friction wedge suspensions are the most widely used bogies in heavy haul trains. Fiction wedge suspensions play a key role in these wagon systems. This article reviews current techniques in dynamic modelling of friction wedge suspension with various motivations: to improve dynamic models of friction wedge suspensions so as to improve general wagon dynamics simulations; to seek better friction wedge suspension models for wagon stability assessments in complex train systems; to improve the modelling of other friction devices, such as friction draft gear. Relevant theories and friction wedge suspension models developed by using commercial simulation packages and in-house simulation packages are reviewed.

  4. First results from the 'Violin-Mode' tests on an advanced LIGO suspension at MIT

    International Nuclear Information System (INIS)

    Lockerbie, N A; Tokmakov, K V; Carbone, L; Shapiro, B; Bell, A; Strain, K A

    2011-01-01

    This paper describes the first results from 'Violin-Mode' measurements made on the four suspension fibres of a fully suspended 40 kg test mass. These measurements were made at the LIGO lab, Gravitational Wave Observatory test facility, at MIT. Here, an aluminium-alloy (dummy) test mass, simulating an advanced LIGO (Laser Interferometer Gravitational Wave Observatory) test mass/mirror, had been suspended in air from a test suspension by four fused-silica suspension fibres, each measuring 400 μm in diameter x 600 mm long. Violin-Mode measurements were made on these highly tensioned fibres by retrofitting a prototype system of four novel shadow sensors to the test suspension, one per fibre, these sensors having, collectively, a displacement sensitivity of (6.9 ± 1.3) x 10 -11 m (rms) Hz -1/2 , at 500 Hz, over a measuring span of ±0.1 mm. Violin-Mode fundamental resonances were detected in all four fibres: with frequencies ∼ 485 Hz when the test mass was supported lightly from below, and at ∼500 Hz when it was fully suspended. In the latter case the Violin-Mode detection took place whilst the test mass, together with its suspension fibres, was undergoing relatively large-amplitude 'pendulum-mode' motion, at ∼0.6 Hz. This motion was measured to have a peak-peak amplitude at one of the suspension fibres of up to ∼140 μm (35 μm, rms) the shadow sensors each having subsidiary outputs for monitoring such low-frequency, large amplitude, motion. Under fully suspended conditions, a calibrated Violin-Mode 'free-oscillation' amplitude of 430 ± 20 picometres, rms, was measured at 500.875 Hz, in the same suspension fibre which was found to be undergoing, simultaneously, the ∼140 μm peak-peak motion. Over the bandwidth monitored (dc to 3.2 kHz), Violin-Mode harmonics up to the sixth were recorded in an evoked response. It was concluded that the prototype system had demonstrated amply its practical viability as a detector of Violin-Mode resonances in the test

  5. First results from the ‘Violin-Mode’ tests on an advanced LIGO suspension at MIT

    Science.gov (United States)

    Lockerbie, N. A.; Carbone, L.; Shapiro, B.; Tokmakov, K. V.; Bell, A.; Strain, K. A.

    2011-12-01

    This paper describes the first results from ‘Violin-Mode’ measurements made on the four suspension fibres of a fully suspended 40 kg test mass. These measurements were made at the LIGO lab, Gravitational Wave Observatory test facility, at MIT. Here, an aluminium-alloy (dummy) test mass, simulating an advanced LIGO (Laser Interferometer Gravitational Wave Observatory) test mass/mirror, had been suspended in air from a test suspension by four fused-silica suspension fibres, each measuring 400 µm in diameter × 600 mm long. Violin-Mode measurements were made on these highly tensioned fibres by retrofitting a prototype system of four novel shadow sensors to the test suspension, one per fibre, these sensors having, collectively, a displacement sensitivity of (6.9 ± 1.3) × 10-11 m (rms) Hz-1/2, at 500 Hz, over a measuring span of ±0.1 mm. Violin-Mode fundamental resonances were detected in all four fibres: with frequencies ˜ 485 Hz when the test mass was supported lightly from below, and at ˜500 Hz when it was fully suspended. In the latter case the Violin-Mode detection took place whilst the test mass, together with its suspension fibres, was undergoing relatively large-amplitude ‘pendulum-mode’ motion, at ˜0.6 Hz. This motion was measured to have a peak-peak amplitude at one of the suspension fibres of up to ˜140 µm (35 µm, rms)—the shadow sensors each having subsidiary outputs for monitoring such low-frequency, large amplitude, motion. Under fully suspended conditions, a calibrated Violin-Mode ‘free-oscillation’ amplitude of 430 ± 20 picometres, rms, was measured at 500.875 Hz, in the same suspension fibre which was found to be undergoing, simultaneously, the ˜140 µm peak-peak motion. Over the bandwidth monitored (dc to 3.2 kHz), Violin-Mode harmonics up to the sixth were recorded in an evoked response. It was concluded that the prototype system had demonstrated amply its practical viability as a detector of Violin-Mode resonances in the

  6. Crust formation in drying colloidal suspensions

    KAUST Repository

    Style, R. W.

    2010-06-30

    During the drying of colloidal suspensions, the desiccation process causes the suspension near the air interface to consolidate into a connected porous matrix or crust. Fluid transport in the porous medium is governed by Darcy\\'s law and the equations of poroelasticity, while the equations of colloid physics govern processes in the suspension. We derive new equations describing this process, including unique boundary conditions coupling the two regions, yielding a moving-boundary model of the concentration and stress profiles during drying. A solution is found for the steady-state growth of a nedimensional crust during constant evaporation rate from the surface. The solution is used to demonstrate the importance of the system boundary conditions on stress profiles and diffusivity in a drying crust. © 2011 The Royal Society.

  7. Anomalous electrical conductivity of nanoscale colloidal suspensions.

    Science.gov (United States)

    Chakraborty, Suman; Padhy, Sourav

    2008-10-28

    The electrical conductivity of colloidal suspensions containing nanoscale conducting particles is nontrivially related to the particle volume fraction and the electrical double layer thickness. Classical electrochemical models, however, tend to grossly overpredict the pertinent effective electrical conductivity values, as compared to those obtained under experimental conditions. We attempt to address this discrepancy by appealing to the complex interconnection between the aggregation kinetics of the nanoscale particles and the electrodynamics within the double layer. In particular, we model the consequent alterations in the effective electrophoretic mobility values of the suspension by addressing the fundamentals of agglomeration-deagglomeration mechanisms through the pertinent variations in the effective particulate dimensions, solid fractions, as well as the equivalent suspension viscosity. The consequent alterations in the electrical conductivity values provide a substantially improved prediction of the corresponding experimental findings and explain the apparent anomalous behavior predicted by the classical theoretical postulates.

  8. Intelligent systems of the vehicles’ suspension

    Science.gov (United States)

    Yurlin, D.

    2018-02-01

    The article is devoted to the current condition of car’s active suspension system. It presents the tendencies in development of the active systems of suspension system, adjustable elements incorporated in them and the companies succeeded in designing such systems. It also mirrors the problem of impact of active systems on car’s safety and their importance for the driver. Advantages and disadvantages of the most common types of active elements are being described, analyzed and compared. The author concludes about the perspectives of these systems’ development.

  9. Unifying Suspension and Granular flows near Jamming

    Directory of Open Access Journals (Sweden)

    DeGiuli Eric

    2017-01-01

    Full Text Available Rheological properties of dense flows of hard particles are singular as one approaches the jamming threshold where flow ceases, both for granular flows dominated by inertia, and for over-damped suspensions. Concomitantly, the lengthscale characterizing velocity correlations appears to diverge at jamming. Here we review a theoretical framework that gives a scaling description of stationary flows of frictionless particles. Our analysis applies both to suspensions and inertial flows of hard particles. We report numerical results in support of the theory, and show the phase diagram that results when friction is added, delineating the regime of validity of the frictionless theory.

  10. Suspension Hydrogen Reduction of Iron Oxide Concentrates

    Energy Technology Data Exchange (ETDEWEB)

    H.Y. Sohn

    2008-03-31

    The objective of the project is to develop a new ironmaking technology based on hydrogen and fine iron oxide concentrates in a suspension reduction process. The ultimate objective of the new technology is to replace the blast furnace and to drastically reduce CO2 emissions in the steel industry. The goals of this phase of development are; the performance of detailed material and energy balances, thermochemical and equilibrium calculations for sulfur and phosphorus impurities, the determination of the complete kinetics of hydrogen reduction and bench-scale testing of the suspension reduction process using a large laboratory flash reactor.

  11. Carbon nanotube suspensions, dispersions, & composites

    Science.gov (United States)

    Simmons, Trevor John

    Carbon Nanotubes (CNTs) are amazing structures that hold the potential to revolutionize many areas of scientific research. CNTs can be behave both as semiconductors and metals, can be grown in highly ordered arrays and patterns or in random orientation, and can be comprised of one graphene cylinder (single wall nanotube, SWNT) or several concentric graphene cylinders (multi-wall nanotube, MWNT). Although these structures are usually only a few nanometers wide, they can be grown up to centimeter lengths, and in massive quantities. CNTs can be produced in a variety of processes ranging from repeated combustion of organic material such as dried grass, arc-discharge with graphite electrodes, laser ablation of a graphitic target, to sophisticated chemical vapor deposition (CVD) techniques. CNTs are stronger than steel but lighter than aluminum, and can be more conductive than copper or semiconducting like silicon. This variety of properties has been matched by the wide variety of applications that have been developed for CNTs. Many of these applications have been limited by the inability of researchers to tame these structures, and incorporating CNTs into existing technologies can be exceedingly difficult and prohibitively expensive. It is therefore the aim of the current study to develop strategies for the solution processing and deposition of CNTs and CNT-composites, which will enable the use of CNTs in existing and emerging technologies. CNTs are not easily suspended in polar solvents and are extremely hydrophobic materials, which has limited much of the solution processing to organic solvents, which also cannot afford high quality dispersions of CNTs. The current study has developed a variety of aqueous CNT solutions that employ surfactants, water-soluble polymers, or both to create suspensions of CNTs. These CNT 'ink' solutions were deposited with a variety of techniques that have afforded many interesting structures, both randomly oriented as well as highly

  12. Rheological Properties of Aqueous Nanometric Alumina Suspensions

    Energy Technology Data Exchange (ETDEWEB)

    Li, Chuanping [Iowa State Univ., Ames, IA (United States)

    2004-01-01

    Colloidal processing is an effective and reliable approach in the fabrication of the advanced ceramic products. Successful colloidal processing of fine ceramic powders requires accurate control of the rheological properties. The accurate control relies on the understanding the influences of various colloidal parameters on the rheological properties. Almost all research done on the rheology paid less attention to the interactions of particle and solvent. However, the interactions of the particles are usually built up through the media in which the particles are suspended. Therefore, interactions of the particle with the media, the adsorbed layers on the particle surface, and chemical and physical properties of media themselves must influence the rheology of the suspension, especially for the dense suspensions containing nanosized particles. Relatively little research work has been reported in this area. This thesis addresses the rheological properties of nanometric alumina aqueous suspensions, and paying more attention to the interactions between particle and solvent, which in turn influence the particle-particle interactions. Dense nanometric alumina aqueous suspensions with low viscosity were achieved by environmentally-benign fructose additives. The rheology of nanometric alumina aqueous suspensions and its variation with the particle volume fraction and concentration of fructose were explored by rheometry. The adsorptions of solute (fructose) and solvent (water) on the nanometric alumina particle surfaces were measured and analyzed by TG/DSC, TOC, and NMR techniques. The mobility of water molecules in the suspensions and its variation with particle volume fractions and fructose additive were determined by the 17O NMR relaxation method. The interactions between the nanometric alumina particles in water and fructose solutions were investigated by AFM. The results indicated that a large number of water layers were physically bound on the particles

  13. Flow Electrification in Nonaqueous Colloidal Suspensions, studied with Video Microscopy

    NARCIS (Netherlands)

    Tolpekin, V.A.; van den Ende, Henricus T.M.; Duits, Michael H.G.; Mellema, J.

    2004-01-01

    Flow electrification in nonaqueous suspensions has been scarcely reported in the literature but can significantly affect colloidal stability and (phase) behavior, perhaps even without being recognized. We have observed it in shear flow experiments on concentrated binary suspensions of hydrophobized

  14. Etude thermique expérimentale des suspensions non newtoniennes ...

    African Journals Online (AJOL)

    Newtonian and pseudoplastic suspensions flowing in a horizontal pipe with variable geometry. These suspensions are composed of large hard spheres in a solution of Tylose. It helped to highlight the limits of the development of the thermal boundary ...

  15. Acid-Fast Staining and Petroff Hausser Chamber Counting of Mycobacterial Cells in Liquid Suspension

    OpenAIRE

    Treuer, Robin; Haydel, Shelley E.

    2011-01-01

    Accurate and rapid cell counts of mycobacterial species in culture are difficult to obtain. Here, a method using modified Kinyoun acid-fast staining was adapted for use with a Petroff-Hausser sperm and bacteria cell counting chamber by using a liquid suspension staining technique. Cell counts obtained by this method were compared to viable cell counts by agar plate counting, revealing accurate correlation.

  16. An engineering characterisation of shaken bioreactors: flow, mixing and suspension dynamics

    OpenAIRE

    Rodriguez, G.

    2017-01-01

    The thesis describes an experimental investigation of the flow, mixing and suspension dynamics in cylindrical orbitally shaken bioreactors (OSRs). Amongst the plethora of bioreactor types and geometries available for cell culture, the OSR is ubiquitous in bioprocess research and development. Offering a well defined liquid-gas interface, high throughput potential and experimental flexibility, it is the vessel of choice in early bioprocess research, either as microtiter-plates, Erlenmeyer flask...

  17. Tangential Ultrafiltration of Aqueous "Saccharomyces Cerevisiae" Suspensions

    Science.gov (United States)

    Silva, Carlos M.; Neves, Patricia S.; Da Silva, Francisco A.; Xavier, Ana M. R. B.; Eusebio, M. F. J.

    2008-01-01

    Experimental work on ultrafiltration is presented to illustrate the practical and theoretical principles of this separation technique. The laboratory exercise comprises experiments with pure water and with aqueous "Saccharomyces cerevisiae" (from commercial Baker's yeast) suspensions. With this work students detect the characteristic phenomena…

  18. Nucleonic coal detector with independent, hydropneumatic suspension

    Science.gov (United States)

    Jones, E. W.; Handy, K.

    1977-01-01

    The design of a nucleonic, coal interface detector which measures the depth of coal on the roof and floor of a coal mine is presented. The nucleonic source and the nucleonic detector are on independent hydropneumatic suspensions to reduce the measurement errors due to air gap.

  19. Aeroelastic Stability of Suspension Bridges using CFD

    DEFF Research Database (Denmark)

    Stærdahl, Jesper Winther; Sørensen, Niels; Nielsen, Søren R.K.

    2007-01-01

    In recent years large span suspension bridges with very thin and slender profiles have been built without proportional increasing torsional and bending stiffness. As a consequence large deformations at the mid-span can occur with risk of aeroelastic instability and structural failure. Analysis of...

  20. 21 CFR 520.1289 - Lufenuron suspension.

    Science.gov (United States)

    2010-04-01

    ...) Limitations. For oral use in cats 6 weeks of age or older, once a month, mixed with food. Administer in... Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS § 520.1289 Lufenuron suspension. (a...

  1. 21 CFR 520.1806 - Piperazine suspension.

    Science.gov (United States)

    2010-04-01

    ... Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS § 520.1806 Piperazine suspension..., reworm at monthly intervals. Not for use in unweaned pups or animals less than 3 weeks of age. [70 FR...

  2. 21 CFR 520.1630 - Oxfendazole suspension.

    Science.gov (United States)

    2010-04-01

    ... Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS § 520.1630 Oxfendazole suspension... use in female dairy cattle of breeding age. [55 FR 46943, Nov. 8, 1990, as amended at 56 FR 8710, Mar...

  3. Absorption Spectra of Gold Nanoparticle Suspensions

    Science.gov (United States)

    Anan'eva, M. V.; Nurmukhametov, D. R.; Zverev, A. S.; Nelyubina, N. V.; Zvekov, A. A.; Russakov, D. M.; Kalenskii, A. V.; Eremenko, A. N.

    2018-02-01

    Three gold nanoparticle suspensions are obtained, and mean radii in distributions - (6.1 ± 0.2), (11.9 ± 0.3), and (17.3 ± 0.7) nm - are determined by the transmission electron microscopy method. The optical absorption spectra of suspensions are obtained and studied. Calculation of spectral dependences of the absorption index of suspensions at values of the gold complex refractive index taken from the literature showed a significant deviation of experimental and calculated data in the region of 450-800 nm. Spectral dependences of the absorption of suspensions are simulated within the framework of the Mie-Drude theory taking into account the interband absorption in the form of an additional term in the imaginary part of the dielectric permittivity of the Gaussian type. It is shown that to quantify the spectral dependences in the region of the plasmon absorption band of nanoparticles, correction of the parameters of the interband absorption is necessary in addition to the increase of the relaxation parameter of the Drude theory. Spectral dependences of the dielectric permittivity of gold in nanodimensional state are refined from the solution of the inverse problem. The results of the present work are important for predicting the special features of operation of photonic devices and optical detonators based on gold nanoparticles.

  4. "Restorative Practices" Offer Alternatives to Suspension

    Science.gov (United States)

    Shah, Nirvi

    2012-01-01

    At City Springs and many other schools across the country, restorative practices are about holding students accountable and getting them to right a wrong. The approach is getting more notice than ever as criticism grows of zero-tolerance disciplinary policies that often require out-of-school suspension and expulsion. Educators are turning to…

  5. 45 CFR 1210.3-3 - Suspension.

    Science.gov (United States)

    2010-10-01

    ... 45 Public Welfare 4 2010-10-01 2010-10-01 false Suspension. 1210.3-3 Section 1210.3-3 Public Welfare Regulations Relating to Public Welfare (Continued) CORPORATION FOR NATIONAL AND COMMUNITY SERVICE VISTA TRAINEE DESELECTION AND VOLUNTEER EARLY TERMINATION PROCEDURES VISTA Volunteer Early Termination...

  6. Formation of Aqueous Suspensions of Fullerenes

    Science.gov (United States)

    Colloidal suspensions of C60, C70 and a derivative of C60, PCBM ([6,6]-Phenyl C61-butyric acid methyl ester) were produced by extended mixing in water. We examined the contribution of background solution chemistry (pH, ionic strength) on the formation kinetics of colloidal suspe...

  7. Pair-correlations in swimmer suspensions

    Science.gov (United States)

    Nambiar, Sankalp; Subramanian, Ganesh

    2017-11-01

    Suspensions of rear-actuated swimming microorganisms, such as E.coli, exhibit several interesting phenomena including spontaneous pattern formation above a critical concentration, novel rheological properties, shear-induced concentration banding etc. Explanations based on mean-field theory are only qualitative, since interactions between swimmers are important for typical experimental concentrations. We analytically characterize the hydrodynamic pair-interactions in a quiescent suspension of slender straight swimmers. The pair-correlation, calculated at leading order by integrating the swimmer velocity disturbances along straight trajectories, decays as 1/r2 for r >> L (L being the swimmer size). This allows us to characterize both polar and nematic correlations in an interacting swimmer suspension. In the absence of correlations, the velocity covariance asymptotes from a constant for r > L, the latter being characteristic of a suspension of non-interacting point force-dipoles. On including correlations, the slow decay of the pair-orientation correlation leads to an additional contribution to the velocity covariance that diverges logarithmically with system size.

  8. 78 FR 57523 - Suspension of Community Eligibility

    Science.gov (United States)

    2013-09-19

    ... scheduled for suspension on the effective dates listed within this rule because of noncompliance with the... participation status of a community can be obtained from FEMA's Community Status Book (CSB). The CSB is... made, this final rule may take effect within less than 30 days. National Environmental Policy Act. This...

  9. 78 FR 65212 - Suspension of Community Eligibility

    Science.gov (United States)

    2013-10-31

    ... scheduled for suspension on the effective dates listed within this rule because of noncompliance with the... participation status of a community can be obtained from FEMA's Community Status Book (CSB). The CSB is... made, this final rule may take effect within less than 30 days. National Environmental Policy Act. This...

  10. 78 FR 51076 - Suspension of Community Eligibility

    Science.gov (United States)

    2013-08-20

    ... scheduled for suspension on the effective dates listed within this rule because of noncompliance with the... participation status of a community can be obtained from FEMA's Community Status Book (CSB). The CSB is... made, this final rule may take effect within less than 30 days. National Environmental Policy Act. This...

  11. 78 FR 57525 - Suspension of Community Eligibility

    Science.gov (United States)

    2013-09-19

    ... scheduled for suspension on the effective dates listed within this rule because of noncompliance with the... participation status of a community can be obtained from FEMA's Community Status Book (CSB). The CSB is... made, this final rule may take effect within less than 30 days. National Environmental Policy Act. This...

  12. 78 FR 5734 - Suspension of Community Eligibility

    Science.gov (United States)

    2013-01-28

    ... scheduled for suspension on the effective dates listed within this rule because of noncompliance with the... participation status of a community can be obtained from FEMA's Community Status Book (CSB). The CSB is... made, this final rule may take effect within less than 30 days. National Environmental Policy Act. This...

  13. 78 FR 45460 - Suspension of Community Eligibility

    Science.gov (United States)

    2013-07-29

    ... scheduled for suspension on the effective dates listed within this rule because of noncompliance with the... participation status of a community can be obtained from FEMA's Community Status Book (CSB). The CSB is... made, this final rule may take effect within less than 30 days. National Environmental Policy Act. This...

  14. 78 FR 70235 - Suspension of Community Eligibility

    Science.gov (United States)

    2013-11-25

    ... scheduled for suspension on the effective dates listed within this rule because of noncompliance with the... participation status of a community can be obtained from FEMA's Community Status Book (CSB). The CSB is... made, this final rule may take effect within less than 30 days. National Environmental Policy Act. This...

  15. 78 FR 65882 - Suspension of Community Eligibility

    Science.gov (United States)

    2013-11-04

    ... scheduled for suspension on the effective dates listed within this rule because of noncompliance with the... participation status of a community can be obtained from FEMA's Community Status Book (CSB). The CSB is... made, this final rule may take effect within less than 30 days. National Environmental Policy Act. This...

  16. Frequency of morning ketosis after overnight insulin suspension using an automated nocturnal predictive low glucose suspend system.

    Science.gov (United States)

    Beck, Roy W; Raghinaru, Dan; Wadwa, R Paul; Chase, H Peter; Maahs, David M; Buckingham, Bruce A

    2014-01-01

    To assess the effect of overnight insulin pump suspension in an automated predictive low glucose suspend system on morning blood glucose and ketone levels in an attempt to determine whether routine measurement of ketone levels is useful when a closed-loop system that suspends insulin delivery overnight is being used. Data from an in-home randomized trial of 45 individuals with type 1 diabetes (age range 15-45 years) were analyzed, evaluating an automated predictive low glucose pump suspension system in which blood glucose, blood ketone, and urine ketone levels were measured on 1,954 mornings. One or more pump suspensions occurred during 744 of the 977 intervention nights (76%). The morning blood ketone level was ≥0.6 mmol/L after 11 of the 744 nights (1.5%) during which a pump suspension occurred and 2 of the 233 nights (0.9%) during which there was no suspension compared with 11 of 977 control nights (1.1%). The morning blood ketone level was ≥0.6 mmol/L after only 2 of 159 nights (1.3%) with a pump suspension exceeding 2 h. Morning fasting blood glucose level was not a good predictor of the presence of blood ketones. Routine measurement of blood or urine ketones during use of an automated pump suspension system using continuous glucose monitoring, whether threshold based or predictive, is not necessary. Recommendations for checking ketone levels should be no different when a patient is using a system with automated insulin suspension than it is for conventional diabetes self-management.

  17. 48 CFR 52.242-14 - Suspension of Work.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 2 2010-10-01 2010-10-01 false Suspension of Work. 52.242... Suspension of Work. As prescribed in 42.1305(a), insert the following clause in solicitations and contracts when a fixed-price construction or architect-engineer contract is contemplated: Suspension of Work (APR...

  18. 21 CFR 529.400 - Chlorhexidine tablets and suspension.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Chlorhexidine tablets and suspension. 529.400... Chlorhexidine tablets and suspension. (a) Specification. Each tablet and each 28-milliliter syringe of suspension contain 1 gram of chlorhexidine dihydrochloride.1 1 These conditions are NAS/NRC reviewed and...

  19. Behaviour of humic-bentonite aggregates in diluted suspensions ...

    African Journals Online (AJOL)

    Formation and disaggregation of micron-size aggregates in a diluted suspension made up of HSs and bentonite (B) were studied by tracing distribution of aggregate sizes and their counts in freshly prepared and aged suspensions, and at high (10 000) and low (1.0) [HS]/[B] ratios. Diluted HSB suspensions are unstable ...

  20. Fuzzy logic control of vehicle suspensions with dry friction nonlinearity

    Indian Academy of Sciences (India)

    We design and investigate the performance of fuzzy logic-controlled (FLC) active suspensions on a nonlinear vehicle model with four degrees of freedom, without causing any degeneration in suspension working limits. Force actuators were mounted parallel to the suspensions. In this new approach, linear combinations of ...