WorldWideScience

Sample records for protoplast culture techniques

  1. Dendrobium protoplast co-culture promotes phytochemical assemblage in vitro.

    Science.gov (United States)

    Thomas, Abitha; Pujari, Ipsita; Shetty, Vasudeep; Joshi, Manjunath B; Rai, Padmalatha S; Satyamoorthy, Kapaettu; Babu, Vidhu Sankar

    2017-07-01

    The present study is intended to analyze the occurrence of potent, low produce, naturally occurring stilbenes in protoplasts of wild species and hybrids of Dendrobium. The wild species selected for the study was Dendrobium ovatum, endemic to Western Ghats of India. Protoplasts were isolated from leaves and tepal tissues of all the species and were cultured purely to generate homofusants and cross-cultured to raise heterofusants. Phytochemical composition of protoplast culture with atypical and pure microcolonies was performed using mass spectrometry. Enzyme cocktail of 4% pectinase together with 2% cellulase displayed the highest competence for protoplast isolations. Maximum protoplast density of 30.11 × 10 4 /ml was obtained from D. ovatum leaves in 2 h. Subcellular features such as the presence of partially formed cell wall, the position of the nucleus, chloroplast density, colony existence, and integrity of the plasma membrane were analyzed. Among the pure and cross-cultured protoplasts, the number of heterofusants and homofusants formed were enumerated. The spectral feature extraction of the mass spectrometry indicated the presence of five phenolic marker compounds, viz., tristin, confusarin, gigantol, moscatilin, and resveratrol, some of them in pure and others in assorted protoplast cultures raised from Dendrobium leaves and tepals. The study demonstrated that protoplast fusion technique enabled phytochemical assemblage in vitro as stilbenes tend to get restricted either in a tissue or species specific manner. This is the first report showing the presence of resveratrol, moscatilin, tristin, gigantol, and confusarin in wild and hybrid species from cultured Dendrobium protoplasts in vitro.

  2. 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.

  3. High efficiency protoplast isolation from in vitro cultures and hairy ...

    African Journals Online (AJOL)

    In vitro cultures of the medicinal plant Maesa lanceolata were established to enable the cultivation of plant material for the production of protoplasts. Callus cultures were initiated using leaves collected from shoot cultures and the root tips from hairy root cultures obtained upon Agrobacterium rhizogenes transformation.

  4. Isolation and culture of protoplast from leaves of Lactuca sativa

    Directory of Open Access Journals (Sweden)

    Witool Chaipakdee

    2007-07-01

    Full Text Available Protoplasts were isolated from leaves of lettuce (Lactuca sativa L. seedlings after in vitro germination for 25, 30, 40 and 50 days. The leaves were stripped and incubated in various combinations of cellulase and pectinase. Protoplasts were cultured on MS medium containing various kinds and concentrations of plant growth regulators in different culture systems including liquid media, hanging, drop culture and solid media. Results revealed that the highest number of viable protoplasts, 14.1x105 cells per gram of fresh weight, was obtained from 30 day-old leaves of lettuce seedlings and isolated by using 2% cellulase in combination with 1% pectinase. Liquid MS medium supplemented with 0.5 mg/l NAA and 0.5 mg/l BA promoted the highest cell division up to 17.67%. First division of protoplasts was observed at 4 days after culture and microcolony formation occurred at the 4th week after culturing. Unfortunately, neither callus formation nor plantlet regeneration were obtained.

  5. Isolation and culture of leaf protoplasts from Tunisian grapes

    Directory of Open Access Journals (Sweden)

    Ahmed Mliki

    2003-09-01

    Full Text Available Experimental conditions for leaf protoplast isolation and culture were optimised for in vitro plants deriving from shoot culture of two Tunisian grape varieties, Sakasly and Muscat d’Alexandrie (Vitis vinifera L.. The best yields were obtained from leaves of 4 to 5 weeks old in vitro plants, digested for 13 hours under 25 rpm agitation with an enzymatic mixture containing 0.25 % cellulase of Aspergillus niger, 0.25 % cellulase of Penicillium funiculosum, 0.5 % cellulysin of Trichoderma viridae, and 0.2 % macerozyme R-10 of Rhizopus sp. More than 50 % of the purified protoplasts had a diameter of 30-40 μm and were rich in chloroplasts. Best aptitude for cell division was found in protoplasts immobilised in sodium alginate layers at a density of 0.5x106 cell/ml, cultivated in CPW-13 medium containing 4 mg/l of NOA and 0.88 mg/l of TDZ. The variety Muscat d’Alexandrie gave better yield whereas Sakasly showed better cell division rates. Formation of micro and macrocallus have been obtained, but the oxidation of the medium has to be solved in order to promote plant regeneration.

  6. Can protoplast production from in vitro cultured shoots of Tanacetum vary during the season?

    Directory of Open Access Journals (Sweden)

    M. KESKITALO

    2008-12-01

    Full Text Available Two different experiments were carried out to study the production of protoplasts and the variation of protoplast yield from in vitro cultured shoot tips of tansy (Tanacetum vulgare L. and pyrethrum (Tanacetum cinerariifolium (Trevir. Schiltz-Bip. In the first experiment, light had more pronouced effect for tansy than for pyrethrum. When the donor tissues of tansy were cultured under high light intensity the leaves contained anthocyanin and became brown during enzyme maceration. In contrast, donor tissues cultured under low light intensity produced leaves without anthocyanin. Depending on the light intensity of donor tissues, on average 5.8 - 6.8 x 106 and 3.4 - 4.3 x 106 protoplasts were isolated from one gram of mesophyll leaves of tansy and pyrethrum, respectively. In the second experiment, the production of protoplasts from tansy and pyrethrum varied seasonally. The most successful season for the production of protoplasts from in vitro cultured shoot tips was between December and April, when also the highest number of protoplasts could be isolated. It was not possible to state whether Tanacetum species have rhythms, which could cause physiological or chemical changes for the in vitro grown shoot tips. However, some external or internal, possible seasonal-dependent stimuli may have caused variation in the number of protoplasts isolated from tansy and pyrethrum and favoured protoplast production during winter and spring.

  7. Secretory production of cell wall components by Saccharomyces cerevisiae protoplasts in static liquid culture.

    Science.gov (United States)

    Aoyagi, Hideki; Ishizaka, Mikiko; Tanaka, Hideo

    2012-04-01

    When protoplasts of Saccharomyces cerevisiae T7 and IFO 0309 are cultured in a static liquid culture at 2.5 × 10(6) protoplasts/ml, cell wall regeneration does not occur and cell wall components (CWC) are released into the culture broth. By using a specialized fluorometer, the concentrations of CWC could be measured on the basis of the fluorescence intensity of the CWC after staining with Fluostain I. The inoculum concentration, pH, and osmotic pressure of the medium were important factors for the production of CWC in culture. Under optimal culture conditions, S. cerevisiae T7 protoplasts produced 0.91 mg/ml CWC after 24 h. The CWC induced the tumor necrosis factor-α production about 1.3 times higher than that of the commercially available β-1,3/1,6-glucan from baker's yeast cells.

  8. Factors affecting callus and protoplast production and regeneration of plants from garlic tissue cultures

    International Nuclear Information System (INIS)

    Al-Safadi, B.; Nabulsi, I.

    2001-08-01

    Five cultivars of garlic, two explants, six callusing media, six regeneration media, two kinds of light and several doses of gamma irradiation were used to determine the best conditions for callus induction and plant regeneration from garlic tissue cultures. Also, some experiments were conducted to study the possibility to isolate protoplast and regenerate plants. The experiment showed that medium MS9 was good for regenerating plant directly from basal plate without going through callus phase. ANOVA exhibited significant differences among used cultivars in their ability to form callus. No significant difference was observed between 16 hr light and complete darkness in callus growth. However, appearance of callus was generally better on darkness. Cultivar varied in their ability to regenerate and interaction between cultivars and media was observed. Cultivar kisswany was the best in regeneration (38%) and medium MS47 was the best among used media (35%). Light type played a significant role in regeneration of plants where red light was much better than white light in inducing regeneration (68% vs 36%). ANOVA revealed significant effect of low doses of gamma irradiation on stimulation regeneration of plant whereas high doses prevented regeneration. Many experiments were conducted to isolate protoplast and regenerate plants. The best method for culturing was the droplet and the best conditions for incubation were complete darkness at 25 Degreed centigrade. This lead to formation of cell wall but no cell division was observed (author)

  9. Soft material-based microculture system having air permeable cover sheet for the protoplast culture of Nicotiana tabacum.

    Science.gov (United States)

    Ju, Jong Il; Ko, Jung-Moon; Kim, So Hyeon; Baek, Ju Yeoul; Cha, Hyeon-Cheol; Lee, Sang Hoon

    2006-08-01

    In plant cell culture, the delivery of nutrition and gas (mainly oxygen) to the cells is the most important factor for viability. In this paper, we propose a polydimethylsiloxane (PDMS)-based microculture system that is designed to have good aeration. PDMS is known to have excellent air permeability, and through the experimental method, we investigated the relation between the degree of air delivery and the thickness of the PDMS sheet covering the culture chamber. We determined the proper thickness of the cover sheet, and cultured protoplasts of Nicotiana tabacum in a culture chamber covered with a PDMS sheet having thickness of 400 microm. The cells were successfully divided, and lived well inside the culture chamber for 10 days. In addition, protoplasts were cultured inside the culture chambers covered with the cover glass and the PDMS sheet, respectively, and the microcolonies were formed well inside the PDMS covered chamber after 10 days.

  10. Patterns of indole alkaloids synthesis in response to heat shock, 5-azacytidine and Na-butyrate treatment of cultured catharanthus roseus mesophyll protoplasts

    International Nuclear Information System (INIS)

    Saleem, M.; Cutler, A.J.

    1986-01-01

    Alkaloids of C. roseus are in high demand for therapeutic and other reasons. Cultured Catharanthus cells can produce limited quantities of these alkaloids. The authors have found that cultured mesophyll protoplasts in the presence of 14 C-Tryptamine are capable of synthesizing alkaloids. The pattern of alkaloids synthesis changes when protoplasts are subjected to a heat shock at 37 0 C. The heat shocked protoplasts incorporated 33% more 14 C-Tryptamine and produced 3 new types of alkaloids. Treatment of protoplasts with 5-azacytidine, a DNA hypomethylating agent and Na-butyrate which induces hyperacetylation of histones produced qualitative and quantitative changes in the alkaloid pattern. Four new alkaloids following the above treatments were detected by TLC and HPLC of the extracts. It is suggested that the alkaloid pattern of the cultured protoplasts can be altered by treatment with compounds known as regulators of gene expression. Work is in progress to isolate and identify these new alkaloids

  11. Use of protoplast, cell, and shoot tip culture in an elm germ plasm improvement program

    Science.gov (United States)

    R. Daniel Lineberger; M.B. Sticklen; P.M. Pijut; Mark A. Kroggel; C.V.M. Fink; S.C. Domir

    1990-01-01

    An elm germplasm improvement program was established using three distinct approaches: (1) development of protoplast regeneration protocols with the goal of attempting somatic hybridization between Ulmus americana and disease resistant hybrids; (2) evaluation of the extent of somaclonal variation in plants regenerated from protoplasts; and (3)...

  12. Isolation and culture of protoplasts of Côte d'Ivoire's pearl millet ...

    African Journals Online (AJOL)

    SARAH

    2015-08-31

    . Journal of Biology and Chemical Science 8 (5):. 2222-2231. Timbo de Oliveira AL, Davide LC, Pereira Pinto JEB,. Pereira AV, 2010. Protoplast production from. Napier grass and Pearl millet triploid hybrids.Ciens.Agrotec.

  13. Methods for suspension culture, protoplast extraction, and transformation of high-biomass yielding perennial grass Arundo donax.

    Science.gov (United States)

    Pigna, Gaia; Dhillon, Taniya; Dlugosz, Elizabeth M; Yuan, Joshua S; Gorman, Connor; Morandini, Piero; Lenaghan, Scott C; Stewart, C Neal

    2016-12-01

    Arundo donax L. is a promising biofuel feedstock in the Mediterranean region. Despite considerable interest in its genetic improvement, Arundo tissue culture and transformation remains arduous. The authors developed methodologies for cell- and tissue culture and genetic engineering in Arundo. A media screen was conducted, and a suspension culture was established using callus induced from stem axillary bud explants. DBAP medium, containing 9 µM 2,4-D and 4.4 µM BAP, was found to be the most effective medium among those tested for inducing cell suspension cultures, which resulted in a five-fold increase in tissue mass over 14 days. In contrast, CIM medium containing 13 µM 2,4-D, resulted in just a 1.4-fold increase in mass over the same period. Optimized suspension cultures were superior to previously-described solidified medium-based callus culture methods for tissue mass increase. Suspension cultures proved to be very effective for subsequent protoplast isolation. Protoplast electroporation resulted in a 3.3 ± 1.5% transformation efficiency. A dual fluorescent reporter gene vector enabled the direct comparison of the CAMV 35S promoter with the switchgrass ubi2 promoter in single cells of Arundo. The switchgrass ubi2 promoter resulted in noticeably higher reporter gene expression compared with that conferred by the 35S promoter in Arundo. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Protoplasts and plant viruses

    International Nuclear Information System (INIS)

    Murakishi, H.; Lesney, M.S.; Carlson, P.

    1984-01-01

    The use of protoplasts in the study of plant viruses has attracted considerable attention since its inception in the late 1960s. This article is an attempt to assess the current status of protoplasts (primarily) and all cell cultures (in some instances) in studies of virus infection, virus replication, cytopathology, cross-protection, virus resistance, and the use of in vitro methods and genetic engineering to recover virus-resistant plants. These areas of study proved difficult to do entirely with whole plants or plant parts. However, because protoplasts could be synchronously infected with virus, they provided a valuable alternative means of following biochemical and cytological events in relation to the virus growth cycle in a more precise manner than previously possible

  15. Intergenus Protoplast Fusion between Pichia manshurica and Rhodosporidium paludigenum to Increase the Production of Inulinase

    Directory of Open Access Journals (Sweden)

    Wijanarka Wijanarka

    2014-12-01

    Full Text Available The purposes of this study was to identify the optimum concentration of the lytic enzyme Glucanex for protoplast isolation and to conduct fusion for the purpose of increasing inulinase production. The study performs the protoplast fusion technique using Pichia manshurica and Rhodosporidium paludigenum. Protoplast fusion consists of a series of stages: protoplast isolation, protoplast fusion, protoplast regeneration, and analysis of hybrid fusion results. Protoplast isolation and fusion success rate are determined by various factors, including age of the culture, media type, and type of lytic enzymes used. Hybrid results were analyzed using a fungicide as a marker and measuring specific growth rate (μ of the hybrid compared with parental growth rates. Results demonstrated that a concentration of 4 mg/mL of Glucanex produces the greatest number of protoplasts, 7.2 x 1010 (cell/mL for P. manshurica and 8.8 x 1010 (cell/mL for Rh. paludigenum. The results of analysis of hybrid fusions indicate that the study has identified a new fusant, called fusant F4. Fusant F4 is capable of producing the highest inulinase, 0.6892 IU, compared with parentals P. manshurica, 0557 IU, and Rh. paludigenum, 0.3263 IU. Fusant F4 has specific growth rate (μ of 0.3360/h and generation time (g of 2.0629 h.

  16. Effect of X-Rays on Growth Rate of Rose Shoot Cultures and the Ability of Isolated Protoplasts to Form Cell Colonies

    International Nuclear Information System (INIS)

    Moustafa, R. A. K.

    2004-01-01

    The popularity of rose as a garden plant, allied with its use in the production of cut flowers and also as a source of aromatic rose oils, make it one of the most important ornamental crops. Roses, however, have suffered from a narrow genetic base to which only few species have contributed significantly. In vitro culture of plants might facilitate the improvement of rose via the exploitation of somaclonal variation to generate new genetic variability and selection within the variation for desirable traits. The application of mutagens for in vitro cultures, in addition to the induced mutations, may lead to increase the somaclonal variation, thus providing additional variation for selection. On the other hand, plant protoplasts offer exciting possibilities to establish in vitro selection programs based on single cells. Induced variation in isolated protoplasts using mutagen agents may be one mean to select useful mutants. Thus the present experiments were conducted to determine the effect of X-rays on shoot cultures and the isolated protoplasts of rose (Rosa sp.). The materials consisted of the three rose varieties Rosa wichuriana, Paricer charm and Heckenzauber.The applied doses were 0, 10, 20, 30, 40, 50 and 60 Gy. Obtained results indicated that the genotypes differed in their sensitivity to X-rays. Rosa wichuriana seemed to be the most sensitive variety to radiation, where a dose of 20 Gy caused approximately 50% reduction in growth rate of shoot cultures, while the same dose decreased the growth rate of Paricer charm only by 25% and did not affect the growth of Heckenzauber. Results also revealed that the ability of irradiated protoplasts to form cell colonies increased when a dose of 10 Gy was performed. Doses higher than that level caused gradual decreasing in the forming of cell colonies, but however, the protoplasts could form colonies even when a dose of 60 Gy was applied. (Author)

  17. Nuclear RNA quantification in protoplast cell-cycle phases.

    Science.gov (United States)

    Bergounioux, C; Perennes, C; Brown, S C; Gadal, P

    1988-01-01

    Using acridine orange staining and flow cytometry the DNA and RNA levels (arbitrary units) of individual cells may be established. Here, this method has been applied to nuclei isolated from plant protoplasts during culture. The specificity of the technique has been validated for such plant material; ribonuclease markedly reduced nuclear staining without modifying the DNA histogram; ribonuclease inhibitor prevented the action of released cell nucleases; and protoplasts cultivated with actinomycin D did not synthesize RNA. First RNA synthesis was evident 18 h after Petunia hybrida protoplasts had been put into culture. An increase of RNA above a critical level was required for cells to be able to initiate DNA replication from G1, termed G1B. G2 nuclei had an RNA:DNA ratio similar to that of G1 nuclei.

  18. Plant tissue culture techniques

    Directory of Open Access Journals (Sweden)

    Rolf Dieter Illg

    1991-01-01

    Full Text Available Plant cell and tissue culture in a simple fashion refers to techniques which utilize either single plant cells, groups of unorganized cells (callus or organized tissues or organs put in culture, under controlled sterile conditions.

  19. Cellulose biodegradation studies: application of rDNA and protoplast fusion techniques

    International Nuclear Information System (INIS)

    Halos, S.C.; Caday, R.; Claudio, J. University of the Philippines, Diliman, Quezon City . Natural Sciences Research Inst.; Pham, L.J.

    1991-01-01

    A gene library of cellulomonas DNA digested with Sau3AI was constructed in BamHI site of pBr322. About 300 recombinants were screened for endo glucanase (CMC-ase) production using congo red assay technique. Out of seven CMC-ase positive clones, four were stable in E. coli C600. The plasmids were extracted from these stable clones and re transformed to HB101 and were analysed for cellulase production both intracellularly and extra cellularly. The plasmids were re isolated from HB101 clones and analysed for inserts using 23 restriction enzymes, which indicated inserts in the range of 6.8-12.5Kb. Cloning of sub fragments indicated two different fragments of 2.8 Kb and 3.7 Kb showing complete CMC-ase activity. This indicates the presence of isozymes in Cellulomonas and strengthens the reports on mutagenic expression of cellulase family. (author)

  20. Protoplast fusion in Streptomyces: fusions involving ultraviolet-irradiated protoplasts

    International Nuclear Information System (INIS)

    Hopwood, D.A.; Wright, H.M.

    1981-01-01

    Protoplasts of Streptomyces coelicolor showed the same ultraviolet killing kinetics as spores. Irradiated protoplasts gave rise to recombinants when they were fused with unirradiated protoplasts of a strain carrying complementary genetic markers. The decline with u.v. fluence in the capacity of irradiated protoplasts to yield recombinants inheriting individual markers was some six times less steep than the survival of unfused protoplasts; thus, for example, protoplasts reduced to only 0.01% survival still yielded 10% as many recombinants as unirradiated protoplasts. Each of six widely separated markers of the irradiated parent was inherited independently of the others, with a frequency falling exponentially with u.v. fluence. (author)

  1. Radiosensitivity of protoplasts of orange (Citrus sinensis)

    International Nuclear Information System (INIS)

    Goldman, M.H.S.; Ando, A.

    1990-01-01

    Full text: The Radiation Genetics Section of the Centre for Nuclear Energy in Agriculture (CENA), University of Sao Paulo, is utilising both ''in vivo'' and ''in vitro'' methods for mutation induction in Citrus, cv. ''Pera'', aiming at resistance to citrus canker. The experiments carried out so far determined the methodology to isolate protoplasts and their sensitivity to gamma-rays. Regarding the culture of protoplasts from embryogenic callus, the best experimental conditions were: enzymatic digestion for 5 h on a medium containing cellulase (307.6 mg/10 ml), macerozyme (30.3 mg/10 ml), mannitol (328.0 mM) and sucrose (336.2 mM) as osmotic stabilisers. The isolation efficiency of 1.2x10 6 viable protoplasts/g will make it possible to use protoplasts in mutation breeding. To determine radiosensitivity of protoplasts, gamma-irradiation from 60 Co source was conducted 42 h after their isolation. This time interval is recommended because during this period protoplasts will reach the stage prior to or at the first mitotic division. Survivals were determined by metylen-blue dyeing, and the LD 50 was found to be around 37.5 Gy. Any difference compared with other authors might be due to different genotypes used or different methods of calculation of survival. (author)

  2. Early Studies on Protoplast Isolation of Ludisia discolor, A Wild Orchid.

    Science.gov (United States)

    Poobathy, Ranjetta; Zakaria, Rahmad; Hamzah, Syed Mohd Edzham Syed; Subramaniam, Sreeramanan

    2016-11-01

    The terrestrial Ludisia discolor , also referred to as the jewel orchid is prized for the quality of its leaves. L. discolor is known as a medicinal herb and is touted for its heat- and pathogen-resisting qualities. L. discolor is valuable in the production of both flavonoids and anthocyanins, antioxidants that are exalted in the health industry. Plant cell cultures have emerged as alternative sources of anthocyanin production. Plant protoplast cultures are used frequently in transient gene expression studies and in the establishment of callus and cell suspension cultures. Benefits of plant protoplast system include similarity to cells found in plant tissues, reproduction under controlled conditions, and prevention of masking of stress responses to previous handling techniques. A study was conducted to assess the amenability of the stem and leaves of L. discolor to protoplast isolation. The stem and leaf segments were weighed, sliced into thin layers, immersed in a digestion medium, washed and then cultured onto a recovery medium. Results indicated that the production of plant protoplasts from L. discolor may be viewed as an alternative in the generation of cell cultures and ultimately in the production of anthocyanins from the cell cultures.

  3. Transfer of herpes simplex virus thymidine kinase synthesized in bacteria by a high-expression plasmid to tissue culture cells by protoplast fusion

    International Nuclear Information System (INIS)

    Waldman, A.S.; Milman, G.

    1984-01-01

    The introduction of a protein into living tissue culture cells may permit the in vivo study of functions of the protein. The authors have previously described a high-efficiency-expression plasmid, pHETK2, containing the herpes simplex virus type 1 thymidine kinase (TK) gene which, upon temperature induction, causes TK to be synthesized as greater than 4% of the bacterial protein. In this report it is shown that enzymatically active TK was transferred to mouse Ltk- cells by polyethylene glycol-mediated fusion with protoplasts prepared from bacteria containing induced levels of TK. The presence of TK in the Ltk- cells was detected by the incorporation of [ 3 H]thymidine into cell nuclei as measured by autoradiography

  4. Plant regeneration from haploid cell suspension-derived protoplasts of Mediterranean rice (Oryza sativa L. cv. Miara).

    Science.gov (United States)

    Guiderdoni, E; Chaïr, H

    1992-11-01

    More than 750 plants were regenerated from protoplasts isolated from microspore callus-derived cell suspensions of the Mediterranean japonica rice Miara, using a nurse-feeder technique and N6-based culture medium. The mean plating efficiency and the mean regeneration ability of the protocalluses were 0.5% and 49% respectively. Flow cytometric evaluation of the DNA contents of 7 month old-cell and protoplast suspensions showed that they were still haploid. Contrastingly, the DNA contents of leaf cell nuclei of the regenerated protoclones ranged from 1C to 5C including 60% 2C plants. This was consistent with the morphological type and the fertility of the mature plants. These results and the absence of chimeric plants suggest that polyploidization occurred during the early phase of protoplast culture.

  5. Formation and cell wall regeneration of protoplasts from Schizophyllum commune

    NARCIS (Netherlands)

    de Vries, Onno Minne Hotze

    1974-01-01

    Osmotically sensitive protoplasts were released from the mycelium of the basidiomycete Schizophyllum commune through the action ofan extracellular enzyme preparation isolated from the culture filtrate of Trichoderma viride (recently renamed T. harzianum) grown on hyphal walls of the former organism.

  6. Effects of environmental preconditioning, donor tissue and isolation conditions on tomato (Lycopersicon esculentum Mill. protoplast yield

    Directory of Open Access Journals (Sweden)

    Elżbieta Kuźniak

    2013-12-01

    Full Text Available The effects of soil or in vitro grown plants, pretreatment conditions, donor tissue and isolation procedure on protoplast yield from cotyledons and leaves of tomato cv. 'Perkoz' and 'Zorza' were studied. The highest protoplast yield of 1.5 x 107/g FW was obtained from leaves of in vitro grown plants. Low light intensity during donor plants in vitro culture and dark pretreatment were essential for successful protoplast isolation while cold pretreatment was not. Tissue preplasmolysis prior to transfer to enzyme mixture increased 4-fold the number of isolated protoplasts. Glycine and bovine serum albumin in the isolation medium did not significantly influence the protoplast yield.

  7. Genetic engineering with tobacco protoplasts. [Hybridization by fusion of leaf protoplasts

    Energy Technology Data Exchange (ETDEWEB)

    Smith, H H

    1976-01-01

    Interspecific hybridization by fusion of leaf protoplasts of Nicotiana glauca (GG) and N. langsdorffii (LL) was confirmed and extended. Enzymatic digestion of leaf tissues to obtain protoplats was followed by fusion with the aid of polyethylene glycol. The hybrid calli were selected by their better growth on defined culture media. Mature hybrid plants were identified by their morphology and tumor formation. Cytological examination revealed a range in chromosome numbers from 56 to 64 rather than the amphiploid GGLL number of 42. About 75 percent of the hybrids were fertile. The potential range in combining widely disparate genotypes by somatic cell fusion was demonstrated by fusing tobacco GGLL protoplasts with human HeLa cells. The HeLa nucleus was observed inside the plant protoplasts, thus forming an interkingdom heterokaryon.

  8. Protoplast formation and regeneration in Lactobacillus delbrueckii

    OpenAIRE

    Singhvi, Mamta; Joshi, Dipti; Gaikaiwari, Shalaka; Gokhale, Digambar V.

    2010-01-01

    Method for production and regeneration of Lactobacillus delbrueckii protoplasts are described. The protoplasts were obtained by treatment with a mixture of lysozyme and mutanolysin in protoplast buffer at pH 6.5 with different osmotic stabilizers. The protoplasts were regenerated on deMan, Rogosa and Sharpe (MRS) with various osmotic stabilizers. Maximum protoplast formation was obtained in protoplast buffer with sucrose as an osmotic stabilizer using a combination of lysozyme (1 mg/ml) and m...

  9. Protoplast preparation from monokaryotic mycelium of Pleurotus sajor-caju using lysing enzyme

    International Nuclear Information System (INIS)

    Hassan Hamdani Mutaat; Mat Rasol Awang

    2004-01-01

    The objective of this study was to determine the optimum parameters of the factors influencing protoplast isolation from monokaryotic mycelium of Pleurotus sajor-caju using lysing enzyme from Trichoderma harzianurm. The study was conducted by manipulating the variables of the factors affecting protoplast isolation, such as age of mycelium culture, period for lysing of mycelium, concentration of lysing enzyme and concentration of osmotic stabilizer. The highest protoplast yield of 8.3 x 104 protoplast/ml was achieved when a 3-day P. sajor-caju mycelium, cultured statically, was incubated for 3 hours in a lytic mixture containing 7.5 mg/ml lysing enzyme and 1.2 M ammonium sulfate as osmotic stabilizer. This protoplast yield, however, is insufficient for regeneration and protoplast fusion works. (Author)

  10. Improved efficiency of plant regeneration from protoplasts of eggplant Solanum melongena L.

    Science.gov (United States)

    Guri, A; Izhar, S

    1984-12-01

    Eggplant (Solanum melongena L.) mesophyll protoplasts were obtained from in vitro growing plants of line 410 and cv. 'Classic'. Relatively high (15%) plating efficiency was achieved using petri dishes with alternate quadrants containing reservoir medium (R medium + 1% activated charcoal) and culture medium. Shoot regeneration occurred within 6 weeks following initiation of protoplast culture.

  11. Effects of environmental preconditioning, donor tissue and isolation conditions on tomato (Lycopersicon esculentum Mill.) protoplast yield

    OpenAIRE

    Elżbieta Kuźniak; Marzena Wielanek; Urszula Małolepsza; Henryk Urbaniak

    2013-01-01

    The effects of soil or in vitro grown plants, pretreatment conditions, donor tissue and isolation procedure on protoplast yield from cotyledons and leaves of tomato cv. 'Perkoz' and 'Zorza' were studied. The highest protoplast yield of 1.5 x 107/g FW was obtained from leaves of in vitro grown plants. Low light intensity during donor plants in vitro culture and dark pretreatment were essential for successful protoplast isolation while cold pretreatment was not. Tissue preplasmolysis prior to t...

  12. Plant regeneration from protoplasts of Gentiana straminea Maxim

    Directory of Open Access Journals (Sweden)

    Shi Guomin

    2016-01-01

    Full Text Available A protocol is described for plant regeneration from protoplasts of Gentiana straminea Maxim. via somatic embryogenesis. Protoplasts were isolated from embryogenic calli in an enzyme solution composed of 2% Cellulase Onozuka R-10, 0.5% Macerozyme R-10, 0.5% Hemicellulase, and 0.5 M sorbitol with a yield of 3.0 × 106 protoplasts per gram of fresh weight. Liquid, solid-liquid double layer (sLD and agar-pool (aPL culture systems were used for protoplast culture. The aPL culture was the only method that produced embryogenic, regenerative calli. With aPL culture, the highest frequencies of protoplast cell division and colony formation were 39.6% and 16.9%, respectively, on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D and 0.5 mg/L N6-benzylaminopurine (BA. Microcalli were transferred to solid MS medium containing a reduced concentration of 2,4-D (0.5 mg/L to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg/L BA at a rate of 43.7%.

  13. Bicarbonate utilization by leaf protoplasts from Potamogeton

    International Nuclear Information System (INIS)

    Staal, M.; Elzenga, J.T.M.; Prins, H.B.A.

    1987-01-01

    Leaves from the submerged angiosperm P. lucens are able to assimilate bicarbonate. These leaves behave polarly: during bicarbonate utilization protons (H + ) are excreted by the cells of the lower epidermis, while hydroxyl (OH - ) ions are excreted by the upper epidermal cells. It has been proposed that acidification of the apoplast is a prerequisite for bicarbonate utilization. To test this hypothesis 14 C fixation by protoplasts was determined at different pH values. Also experiments, using the isotopic disequilibrium technique were performed. They showed that at pH values > 8, bicarbonate is a major carbon source for photosynthesis in protoplasts, despite the absence of cell walls and polarity. At pH values around 6, the rate of 14 C-fixation in protoplasts equals that of intact leaves. At pH values > 8, however, intact leaves show a higher rate. From this, and other experiments, the authors conclude that at least 2 processes contribute to bicarbonate utilization in P. lucens leaves: active transport (H + -HCO 3 - symport?) and acidification of the apoplast resulting in the conversion of bicarbonate into CO 2 . Polarity may increase the efficiency of both

  14. Isolation of protoplast from soybean, cowpea, and tobacco and their fusion

    International Nuclear Information System (INIS)

    Irwansyah.

    1988-01-01

    Protoplast were isolated from leaf and callus. Young leaf of 3-4 weeks old plant of soybean T219 and A24, A27, C4, E1, and H6 of cowpeas strains (strains named by Prof. S. Sakamoto, University of Kyoto) were suspended in digestive medium containing cellulase 'Onuzuka' R-10, macerozyme R-10, mannitol, CaCl, and 2 (N-morpholilno) echane sulfonic acid (MES). For soybean leaf, the medium was enriched with driselase and pectolyase Y-23. They were incibated in full darkness at 27 Celcius centigrade by constant shaking at 50 rpm orbitor shaker. Callus wich has been two times resubcultured was suspended in the digestive medium without driselase, CaCl2, and MES and incubated in lowlight intensity by constant shaking at 100 rpm in reciprocal water shaker at 30 celcius centigrade. Leaf protoplast were releasaed in 10-14 h, soybean and tobacco callus protoplast in 3-4 h, and cowpeas callus protoplast in 4-6 h of incubation. Protoplast were collected by centrifugation of 400 g and a thin layer of the suspension was irradiated with ultraviolet light. Fusion was induced with PEG 6000 solution according to Uchimia and fused protoplasts were collected by centrifugation of 200 g. Protoplast were cultured on the medium of Ikeda and Uchimia. On both medium leaf protoplast, irradiated protoplasts and their fused do not regenerate cell wall and all cultured died out within four weeks incubation. Cell wall generation was observed. Regeneration of cell wall observed progessively in mother protoplast from tobacco, cowpea (A27, E1, and H6) and fused protoplast of soybean with tobacco, tobacco with cowpea (C4, E1, and H6), soybean with cowpea (C4) and between cowpea (C4) and cowpea (E1). (author). 25 refs, 4 tabs

  15. Magnetic field exposure stiffens regenerating plant protoplast cell walls.

    Science.gov (United States)

    Haneda, Toshihiko; Fujimura, Yuu; Iino, Masaaki

    2006-02-01

    Single suspension-cultured plant cells (Catharanthus roseus) and their protoplasts were anchored to a glass plate and exposed to a magnetic field of 302 +/- 8 mT for several hours. Compression forces required to produce constant cell deformation were measured parallel to the magnetic field by means of a cantilever-type force sensor. Exposure of intact cells to the magnetic field did not result in any changes within experimental error, while exposure of regenerating protoplasts significantly increased the measured forces and stiffened regenerating protoplasts. The diameters of intact cells or regenerating protoplasts were not changed after exposure to the magnetic field. Measured forces for regenerating protoplasts with and without exposure to the magnetic field increased linearly with incubation time, with these forces being divided into components based on the elasticity of synthesized cell walls and cytoplasm. Cell wall synthesis was also measured using a cell wall-specific fluorescent dye, and no changes were noted after exposure to the magnetic field. Analysis suggested that exposure to the magnetic field roughly tripled the Young's modulus of the newly synthesized cell wall without any lag.

  16. Efficient production of Aschersonia placenta protoplasts for transformation using optimization algorithms.

    Science.gov (United States)

    Wei, Xiuyan; Song, Xinyue; Dong, Dong; Keyhani, Nemat O; Yao, Lindan; Zang, Xiangyun; Dong, Lili; Gu, Zijian; Fu, Delai; Liu, Xingzhong; Qiu, Junzhi; Guan, Xiong

    2016-07-01

    The insect pathogenic fungus Aschersonia placenta is a highly effective pathogen of whiteflies and scale insects. However, few genetic tools are currently available for studying this organism. Here we report on the conditions for the production of transformable A. placenta protoplasts using an optimized protocol based on the response surface method (RSM). Critical parameters for protoplast production were modelled by using a Box-Behnken design (BBD) involving 3 levels of 3 variables that was subsequently tested to verify its ability to predict protoplast production (R(2) = 0.9465). The optimized conditions resulted in the highest yield of protoplasts ((4.41 ± 0.02) × 10(7) cells/mL of culture, mean ± SE) when fungal cells were treated with 26.1 mg/mL of lywallzyme for 4 h of digestion, and subsequently allowed to recover for 64.6 h in 0.7 mol/L NaCl-Tris buffer. The latter was used as an osmotic stabilizer. The yield of protoplasts was approximately 10-fold higher than that of the nonoptimized conditions. Generated protoplasts were transformed with vector PbarGPE containing the bar gene as the selection marker. Transformation efficiency was 300 colonies/(μg DNA·10(7) protoplasts), and integration of the vector DNA was confirmed by PCR. The results show that rational design strategies (RSM and BBD methods) are useful to increase the production of fungal protoplasts for a variety of downstream applications.

  17. Microcolony formation from embryogenic callus-derived protoplasts of oil palm

    Directory of Open Access Journals (Sweden)

    Sompong Te-chato

    2005-07-01

    Full Text Available Embryogenic callus of oil palm induced from young leaves of seedlings DxP was used as initial material for protoplast isolation. Various combinations of cellulase Onozuka RS and macerozyme R-10 were tested. Isolated protoplasts were cultured by various methods in MS medium supplemented with different phytohormones. The results revealed that 2% cellulase RS in combination with 2% macerozyme R-10 (adjusted osmoticum to 0.4 M by manitol yielded the highest number of viable protoplasts (1x107 per gram fresh weight. Dicamba at concentration 2 mg/l with 1 mg/l 6-benzyladenin (BA containing in phytagel semisolidified MS medium promoted the highest division of 2.3-4.0%. First division of the protoplasts was observed at 4 days after culture. Microcolony formation (8-10 cells was seen after three weeks of culture. Unfortunately, neither callus formation nor plantlet regeneration were obtained.

  18. Inoculation Technique for Fungus Cultures

    Science.gov (United States)

    Fusaro, Ramon M.

    1972-01-01

    A plastic straw and wood applicator stick serve as a simple, inexpensive, and disposable inoculation unit for fungal studies. The method gives a uniform and intact inoculum. The technique is especially useful because a large number of agar plates can be inoculated rapidly. Images PMID:5059618

  19. Optimization of Production Conditions for Protoplasts and Polyethylene Glycol-Mediated Transformation of Gaeumannomyces tritici.

    Science.gov (United States)

    Wang, Mei; Zhang, Jie; Wang, Lanying; Han, Lirong; Zhang, Xing; Feng, Juntao

    2018-05-24

    Take-all, caused by Gaeumannomyces tritici , is one of the most important wheat root diseases worldwide, as it results in serious yield losses. In this study, G. tritici was transformed to express the hygromycin B phosphotransferase using a combined protoplast and polyethylene glycol (PEG)-mediated transformation technique. Based on a series of single-factor experimental results, three major factors-temperature, enzyme lysis time, and concentration of the lysing enzyme-were selected as the independent variables, which were optimized using the response surface methodology. A higher protoplast yield of 9.83 × 10⁷ protoplasts/mL was observed, and the protoplast vitality was also high, reaching 96.27% after optimization. Protoplasts were isolated under the optimal conditions, with the highest transformation frequency (46⁻54 transformants/μg DNA). Polymerase chain reaction and Southern blotting detection indicated that the genes of hygromycin phosphotransferase were successfully inserted into the genome of G. tritici . An optimised PEG-mediated protoplast transformation system for G. tritici was established. The techniques and procedures described will lay the foundation for establishing a good mutation library of G. tritici and could be used to transform other fungi.

  20. Isolation and regeneration protoplast of an oil palm pathogen, Ganoderma boninense

    Science.gov (United States)

    Irene, Liza Isaac; Bakar, Farah Diba Abu; Idris, Abu Seman; Murad, Abdul Munir Abdul

    2015-09-01

    Ganoderma boninense is a known cause for basal stem rot (BSR) in oil palm. Thus, to curb the infection towards oil palm, the establishment of protoplast isolation and regeneration protocol is crucial to be studied. This will provide information on the functional genes especially those which leads towards infection and pathogenicity. In this study, a method was outlined to isolated protoplast in G. boninense by manipulating parameters such as mycelium age, concentration of lysing enzyme, and duration of mycelia incubation in lytic solution. The results shows that from 0.1 g of wet weight mycelia, the highest protoplast yield obtained was 5.5 × 108 protoplast/ml using 5th day old culture in a lytic mixture containing 2.0 % of lysing enzyme incubated for 4 hours at 30 °C with agitation of 80-100 rpm. The highest percentage of protoplast regeneration obtained from this study was 0.2 % using CYM medium supplemented with 0.6 M sorbitol. To date, this is the first report of protoplast isolation and regeneration for this phytopathogen.

  1. Cell culture techniques in honey bee research

    Science.gov (United States)

    Cell culture techniques are indispensable in most if not all life science disciplines to date. Wherever cell culture models are lacking scientific development is hampered. Unfortunately this has been and still is the case in honey bee research because permanent honey bee cell lines have not yet been...

  2. Enhancement of yellow pigment production by intraspecific protoplast fusion of Monascus spp. yellow mutant (ade(-)) and white mutant (prototroph).

    Science.gov (United States)

    Klinsupa, Worawan; Phansiri, Salak; Thongpradis, Panida; Yongsmith, Busaba; Pothiratana, Chetsada

    2016-01-10

    To breed industrially useful strains of a slow-growing, yellow pigment producing strain of Monascus sp., protoplasts of Monascus purpureus yellow mutant (ade(-)) and rapid-growing M. purpureus white mutant (prototroph) were fused and fusants were selected on minimal medium (MM). Preliminary conventional protoplast fusion of the two strains was performed and the result showed that only white colonies were detected on MM. It was not able to differentiate the fusants from the white parental prototroph. To solve this problem, the white parental prototroph was thus pretreated with 20mM iodoacetamide (IOA) for cytoplasm inactivation and subsequently taken into protoplast fusion with slow-growing Monascus yellow mutant. Under this development technique, only the fusants, with viable cytoplasm from Monascus yellow mutant (ade(-)), could thus grow on MM, whereas neither IOA pretreated white parental prototroph nor yellow auxotroph (ade(-)) could survive. Fifty-three fusants isolated from yellow colonies obtained through this developed technique were subsequently inoculated on complete medium (MY agar). Fifteen distinguished yellow colonies from their parental yellow mutant were then selected for biochemical, morphological and fermentative properties in cassava starch and soybean flour (SS) broth. Finally, three most stable fusants (F7, F10 and F43) were then selected and compared in rice solid culture. Enhancement of yellow pigment production over the parental yellow auxotroph was found in F7 and F10, while enhanced glucoamylase activity was found in F43. The formation of fusants was further confirmed by monacolin K content, which was intermediate between the two parents (monacolin K-producing yellow auxotroph and non-monacolin K producing white prototroph). Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Identification of protoplast-isolation responsive microRNAs in Citrus reticulata Blanco by high-throughput sequencing.

    Science.gov (United States)

    Xu, Xiaoyong; Xu, Xiaoling; Zhou, Yipeng; Zeng, Shaohua; Kong, Weiwen

    2017-01-01

    Protoplast isolation is a stress-inducing process, during which a variety of physiological and molecular alterations take place. Such stress response affects the expression of totipotency of cultured protoplasts. MicroRNAs (miRNAs) play important roles in plant growth, development and stress responses. However, the underlying mechanism of miRNAs involved in the protoplast totipotency remains unclear. In this study, high-throughput sequencing technology was used to sequence two populations of small RNA from calli and callus-derived protoplasts in Citrus reticulata Blanco. A total of 67 known miRNAs from 35 families and 277 novel miRNAs were identified. Among these miRNAs, 18 known miRNAs and 64 novel miRNAs were identified by differentially expressed miRNAs (DEMs) analysis. The expression patterns of the eight DEMs were verified by qRT-PCR. Target prediction showed most targets of the miRNAs were transcription factors. The expression levels of half targets showed a negative correlation to those of the miRNAs. Furthermore, the physiological analysis showed high levels of antioxidant activities in isolated protoplasts. In short, our results indicated that miRNAs may play important roles in protoplast-isolation response.

  4. Analysis of the effects of cerium on calcium ion in the protoplasts of ...

    African Journals Online (AJOL)

    The laser-scanning confocal microscopy has become a routine technique and indispensable tool for cell biological studies. In this study, the probe Fluo-3 AM was used to research the instantaneous changes of calcium ion (Ca2+) in the protoplasts of Arabidopsis thaliana. The laser-scanning mode of confocal microscope is ...

  5. Polyamine binding to proteins in oat and Petunia protoplasts

    Science.gov (United States)

    Mizrahi, Y.; Applewhite, P. B.; Galston, A. W.

    1989-01-01

    Previous work (A Apelbaum et al. [1988] Plant Physiol 88: 996-998) has demonstrated binding of labeled spermidine (Spd) to a developmentally regulated 18 kilodalton protein in tobacco tissue cultures derived from thin surface layer explants. To assess the general importance of such Spd-protein complexes, we attempted bulk isolation from protoplasts of Petunia and oat (Avena sativa). In Petunia, as in tobacco, fed radioactive Spd is bound to protein, but in oat, Spd is first converted to 1,3,-diaminopropane (DAP), probably by polyamine oxidase action. In oat, binding of DAP to protein depends on age of donor leaf and conditions of illumination and temperature, and the extraction of the DAP-protein complex depends upon buffer and pH. The yield of the DAP-protein complex was maximized by extraction of frozen-thawed protoplasts with a pH 8.8 carbonate buffer containing SDS. Its molecular size, based on Sephacryl column fractionation of ammonium sulfate precipitated material, exceeded 45 kilodaltons. Bound Spd or DAP can be released from their complexes by the action of Pronase, but not DNAse, RNAse, or strong salt solutions, indicating covalent attachment to protein.

  6. Infection of potato mesophyll protoplasts with five plant viruses.

    Science.gov (United States)

    Barker, H; Harrison, B D

    1982-12-01

    Methods are described for preparing potato mesophyll protoplasts that are suitable for infection with inocula of virus nucleoprotein or RNA. The protoplasts could be infected with four sap-transmissible viruses (tobacco mosaic, tobacco rattle, tobacco ringspot and tomato black ring viruses) and with potato leafroll virus, which is not saptransmissible. No differences were observed in ability to infect protoplasts with potato leafroll virus strains differing either in virulence in intact plants or in aphid transmissibility.

  7. Poinsettia protoplasts - a simple, robust and efficient system for transient gene expression studies

    Directory of Open Access Journals (Sweden)

    Pitzschke Andrea

    2012-05-01

    Full Text Available Abstract Background Transient gene expression systems are indispensable tools in molecular biology. Yet, their routine application is limited to few plant species often requiring substantial equipment and facilities. High chloroplast and chlorophyll content may further impede downstream applications of transformed cells from green plant tissue. Results Here, we describe a fast and simple technique for the high-yield isolation and efficient transformation (>70% of mesophyll-derived protoplasts from red leaves of the perennial plant Poinsettia (Euphorbia pulccherrima. In this method no particular growth facilities or expensive equipments are needed. Poinsettia protoplasts display an astonishing robustness and can be employed in a variety of commonly-used downstream applications, such as subcellular localisation (multi-colour fluorescence or promoter activity studies. Due to low abundance of chloroplasts or chromoplasts, problems encountered in other mesophyll-derived protoplast systems (particularly autofluorescence are alleviated. Furthermore, the transgene expression is detectable within 90 minutes of transformation and lasts for several days. Conclusions The simplicity of the isolation and transformation procedure renders Poinsettia protoplasts an attractive system for transient gene expression experiments, including multi-colour fluorescence, subcellular localisation and promoter activity studies. In addition, they offer hitherto unknown possibilities for anthocyan research and industrial applications.

  8. Cell wall regeneration in Bangia atropurpurea (Rhodophyta) protoplasts observed using a mannan-specific carbohydrate-binding module.

    Science.gov (United States)

    Umemoto, Yoshiaki; Araki, Toshiyoshi

    2010-02-01

    The cell wall of the red alga Bangia atropurpurea is composed of three unique polysaccharides (beta-1,4-mannan, beta-1,3-xylan, and porphyran), similar to that in Porphyra. In this study, we visualized beta-mannan in the regenerating cell walls of B. atropurpurea protoplasts by using a fusion protein of a carbohydrate-binding module (CBM) and green fluorescent protein (GFP). A mannan-binding family 27 CBM (CBM27) of beta-1,4-mannanase (Man5C) from Vibrio sp. strain MA-138 was fused to GFP, and the resultant fusion protein (GFP-CBM27) was expressed in Escherichia coli. Native affinity gel electrophoresis revealed that GFP-CBM27 maintained its binding ability to soluble beta-mannans, while normal GFP could not bind to beta-mannans. Protoplasts were isolated from the fronds of B. atropurpurea by using three kinds of bacterial enzymes. The GFP-CBM27 was mixed with protoplasts from different growth stages, and the process of cell wall regeneration was observed by fluorescence microscopy. Some protoplasts began to excrete beta-mannan at certain areas of their cell surface after 12 h of culture. As the protoplast culture progressed, beta-mannans were spread on their entire cell surfaces. The percentages of protoplasts bound to GFP-CBM27 were 3%, 12%, 17%, 29%, and 25% after 12, 24, 36, 48, and 60 h of culture, respectively. Although GFP-CBM27 bound to cells at the initial growth stages, its binding to the mature fronds was not confirmed definitely. This is the first report on the visualization of beta-mannan in regenerating algal cell walls by using a fluorescence-labeled CBM.

  9. Genetic variability in regenerated Metarhizium flavoviride protoplasts

    Directory of Open Access Journals (Sweden)

    Júlia Kuklinsky-Sobral

    2004-03-01

    Full Text Available Protoplast isolation and regeneration were evaluated in two wild-type and two colour mutant strains of Metarhizium flavoviride. Cultivation in liquid medium, followed by mycelium treatment with Novozym 234 in the presence of KCl 0.7M as osmotic stabilizer, produced 5.05 x 10(6 to 1.15 x 10(7x mL-1 protoplasts. The percentage of regeneration ranged from 6.65 to 27.92%. Following protoplast regeneration, one strain produced spontaneously stable morphological variant colonies. Although colonies with altered morphology have been reported in bacteria following protoplast regeneration, this is the first time that the same is described in a filamentous fungus. The original strain and one derived variant were tested for sensitivity to the fungicides benomyl and captan.A formação e regeneração de protoplastos foram avaliadas em duas linhagens selvagens e duas linhagens mutantes para coloração de conídios em Metarhizium flavoviride. O cultivo em meio líquido seguido do tratamento do micélio com Novozym 234 na presença de KCl 0,7 M como estabilizador osmótico, resultou na produção de 5,05´10(6 a 1,15´10(7 protoplastos´mL-1. A porcentagem de regeneração das diferentes linhagens variou de 6,65 a 27,92%. Após a regeneração, uma das linhagens selvagens produziu espontaneamente variantes estáveis, com morfologia alterada. Embora variantes morfológicos já tenham sido observados após regeneração de protoplastos em bactérias, esta parece ser a primeira vez que tal ocorrência é descrita em fungos filamentosos. Um desses variantes, além da linhagem selvagem da qual ele foi originado, foi testado para sensibilidade aos fungicidas benomil e captano.

  10. Proteins synthesized in tobacco mosaic virus infected protoplasts

    NARCIS (Netherlands)

    Huber, R.

    1979-01-01

    The study described here concerns the proteins, synthesized as a result of tobacco mosaic virus (TMV) multiplication in tobacco protoplasts and in cowpea protoplasts. The identification of proteins involved in the TMV infection, for instance in the virus RNA replication, helps to elucidate

  11. Interaction of E. coli DNA with tobacco mesophyll protoplasts

    International Nuclear Information System (INIS)

    Heyn, R.F.

    1975-01-01

    This chapter is part of a dissertation dealing with the interaction of DNA with protoplasts. Having established the length of time during which tobacco mesophyll protoplasts do not synthesize DNA following their isolation, it is important to know the extent of DNA uptake just before the onset of DNA synthesis (and possible integration) and to find optimal conditions for this uptake. Therefore, the association of E. coli DNA with tobacco protoplasts was studied. Care should be taken with the interpretation of ''uptake'' results: adsorption phenomena play a very important role and may do so at the plasmalemma of naked protoplasts. To solve the problems involved, the use of radiation-damaged DNA was attempted. With E. coli DNA possessing a large number of thymine containing pyrimidine dimers, the loss of dimers from DNA recovered from treated protoplasts was tested in order to obtain an indication of ''real'' uptake. The results are reported

  12. Improvement of large quantity breeding method for making radiation breeding efficient and development of cell culture techniques, (3)

    International Nuclear Information System (INIS)

    Hogetsu, Daisuke; Koyama, Motoko; Minami, Harufumi

    1990-01-01

    In the creation of useful mutant plants using cell culture techniques, the examination on the effectiveness of selecting useful mutation at cell level and the possibility of raising the selection efficiency by irradiation was aimed at. The experimental method is described. The young plants which accumulate proline were obtained. The cells which showed the resistance to hydroxyproline also showed the resistance to salt. In the improvement of redifferentiation ability by irradiation, the method of fixing IAA in the tissues of azuki plants was examined. The possibility of examining the change of IAA due to irradiation by microautoradiography was obtained. It is intended to examine the distribution of IAA in the formation of adventitious roots from the epicotyl of azuki plants. In the introduction of cell engineering techniques in radiation breeding, it is the objective to introduce the genes which resist sour rot that Brassica campestris, Brassica napus, Brassica oleracea and so on have by utilizing cell fusion process. The fusion of the reproduction cells of Brassica napus pollens and the cell protoplasts of Brassica campestris was successfully carried out. The possibility of new asymmetric fusion in Brassica napus was shown. (K.I.)

  13. Profound improvements of isolated microspores culture techniques ...

    African Journals Online (AJOL)

    We studied the effects of sampling stages, physical conditions (like temperature), culture conditions, embryo long-distance transportation methodology and plantlet regeneration on isolated microspores from donor plants in field. Results indicated that if microspores were sampled in bud stage instead of blooming stage to ...

  14. Basidiospore and Protoplast Regeneration from Raised Fruiting Bodies of Pathogenic Ganoderma boninense.

    Science.gov (United States)

    Govender, Nisha T; Mahmood, Maziah; Seman, Idris A; Mui-Yun, Wong

    2016-08-26

    Ganoderma boninense, a phytopathogenic white rot fungus had sought minimal genetic characterizations despite huge biotechnological potentials. Thus, efficient collection of fruiting body, basidiospore and protoplast of G. boninense is described. Matured basidiocarp raised under the glasshouse conditions yielded a total of 8.3 × 104 basidiospores/ml using the low speed centrifugation technique. Mycelium aged 3-day-old treated under an incubation period of 3 h in lysing enzyme from Trichoderma harzianum (10 mg/ml) suspended in osmotic stabilizer (0.6 M potassium chloride and 20 mM dipotassium phosphate buffer) yielded the highest number of viable protoplasts (8.9 × 106 single colonies) among all possible combinations tested (regeneration media, age of mycelium, osmotic stabilizer, digestive enzyme and incubation period).

  15. Plant regeneration from protoplasts ofVicia narbonensis via somatic embryogenesis and shoot organogenesis.

    Science.gov (United States)

    Tegeder, M; Kohn, H; Nibbe, M; Schieder, O; Pickardt, T

    1996-11-01

    Protoplasts ofVicia narbonensis isolated from epicotyls and shoot tips of etiolated seedlings were embedded in 1.4% sodium-alginate at a final density of 2.5×10(5) protoplasts/ml and cultivated in Kao and Michayluk-medium containing 0.5 mg/I of each of 2,4- dichlorophenoxyacetic acid, naphthylacetic acid and 6 -benzylaminopurine. A division frequency of 36% and a plating efficiency of 0.40-0.5% were obtained. Six weeks after embedding, protoplast-derived calluses were transferred onto gelrite-solidified Murashige and Skoog-media containing various growth regulators. Regeneration of plants was achieved via two morphologically distinguishable pathways. A two step protocol (initially on medium with a high auxin concentration followed by a culture phase with lowered auxin amount) was used to regenerate somatic embryos, whereas cultivation on medium containing thidiazuron and naphthylacetic acid resulted in shoot morphogenesis. Mature plants were recovered from both somatic embryos as well as from thidiazuron-induced shoots.

  16. Evidence for some signal transduction elements involved in UV-light-dependent responses in parsley protoplasts

    International Nuclear Information System (INIS)

    Frohnmeyer, H.; Bowler, C.; Schäfer, E.

    1997-01-01

    The signalling pathways used by UV-light are largely unknown. Using protoplasts from a heterotrophic parsley (Petroselinum crispum L.) cell culture that exclusively respond to UV-B light between 300 and 350 nm with a fast induction of genes encoding flavonoid biosynthetic enzymes, information was obtained about the UV-light signal transduction pathway for chalcone synthase (CHS) and phenylalanine ammonia-lyase (PAL) gene expression. Pharmacological effectors which influence intracellular calcium levels, calmodulin and the activity of serine/threonine kinases also changed the UV-light-dependent expression of these genes. This evaluation indicated the participation of these components on the UV-B-mediated signal transduction cascade to CHS. In contrast, neither membrane-permeable cyclic GMP nor the tyrosine kinase inhibitor genistein affected CHS or PAL expression. Similar results were obtained in protoplasts, which have been transiently transformed with CHS-promoter/GUS (β-glucuronidase) reporter fusion constructs. The involvement of calcium and calmodulin was further indicated in a cell-free light-responsive in vitro transcription system from evacuolated parsley protoplasts. In conclusion, there is evidence now that components of the UV-light-dependent pathway leading to the CHS-promoter are different from the previously characterized cGMP-dependent pathway to CHS utilized by phytochrome in soybean (Glycine max) and tomato seedlings (Lycopersicon esculentum). (author)

  17. Viral protein synthesis in cowpea mosaic virus infected protoplasts

    International Nuclear Information System (INIS)

    Rottier, P.

    1980-01-01

    Some aspects of cowpea mosaic virus (CPMV) multiplication in cowpea mesophyll protoplasts were studied. The detection and characterization of proteins whose synthesis is induced or is stimulated upon virus infection was performed with the aid of radioactive labelling. (Auth.)

  18. Culture technique of rabbit primary epidermal keratinocytes

    Directory of Open Access Journals (Sweden)

    Marini M

    2012-10-01

    Full Text Available The epidermis is the protective covering outer layer of the mammalian skin. The epidermal cells are stratified squamous epithelia which undergo continuous differentiation of loss and replacement of cells. Ninety per cent of epidermal cells consist of keratinocytes that are found in the basal layer of the stratified epithelium called epidermis. Keratinocytes are responsible for forming tight junctions with the nerves of the skin as well as in the process of wound healing. This article highlights the method of isolation and culture of rabbit primary epidermal keratinocytes in vitro. Approximately 2cm x 2cm oval shaped line was drawn on the dorsum of the rabbit to mark the surgical area. Then, the skin was carefully excised using a surgical blade and the target skin specimens harvested from the rabbits were placed in transport medium comprising of Dulbecco’s Modified Eagle Medium (DMEM and 1% of antibiotic-antimycotic solution. The specimens were transferred into a petri dish containing 70% ethanol and washed for 5 min followed by a wash in 1 x Dulbecco’s Phosphate Buffered Saline (DBPS. Then, the skin specimens were placed in DMEM and minced into small pieces using a scalpel. The minced pieces were placed in a centrifuge tube containing 0.6% Dispase and 1% antibiotic-antimycotic solution overnight at 4°C in a horizontal orientation. The epidermis layer (whitish, semi-transparent was separated from the dermis (pink, opaque, gooey with the aid of curved forceps by fixing the dermis with one pair of forceps while detaching the epidermis with the second pair. The cells were cultured at a density of 4 x 104 cells/cm2 in culture flask at 37°C and 5% CO2. The cell morphology of the keratinocytes was analyzed using inverted microscope.

  19. Proteins synthesized in tobacco mosaic virus infected protoplasts

    International Nuclear Information System (INIS)

    Huber, R.

    1979-01-01

    The author deals with research on the multiplication of tobacco mosaic virus (TMV) in leaf cell protoplasts. An attempt is made to answer three questions: (1) Which proteins are synthesized in TMV infected protoplasts as a result of TMV multiplication. (2) Which of the synthesized proteins are made under the direction of the TMV genome and, if any, which of the proteins are host specific. (3) In which functions are these proteins involved. (Auth.)

  20. The study of cultural objects by nuclear and conventional techniques

    International Nuclear Information System (INIS)

    Palacios, Tulio A.

    2000-01-01

    A survey is given of the techniques that are used at the National Atomic Energy Commission of Argentina for the characterization and study of cultural and archaeological specimens. A short history of these activities is also given. (author)

  1. Application of Hanging Drop Technique for Kidney Tissue Culture.

    Science.gov (United States)

    Wang, Shaohui; Wang, Ximing; Boone, Jasmine; Wie, Jin; Yip, Kay-Pong; Zhang, Jie; Wang, Lei; Liu, Ruisheng

    2017-01-01

    The hanging drop technique is a well-established method used in culture of animal tissues. However, this method has not been used in adult kidney tissue culture yet. This study was to explore the feasibility of using this technique for culturing adult kidney cortex to study the time course of RNA viability in the tubules and vasculature, as well as the tissue structural integrity. In each Petri dish with the plate covered with sterile buffer, a section of mouse renal cortex was cultured within a drop of DMEM culture medium on the inner surface of the lip facing downward. The tissue were then harvested at each specific time points for Real-time PCR analysis and histological studies. The results showed that the mRNA level of most Na+ related transporters and cotransporters were stably maintained within 6 hours in culture, and that the mRNA level of most receptors found in the vasculature and glomeruli were stably maintained for up to 9 days in culture. Paraffin sections of the cultured renal cortex indicated that the tubules began to lose tubular integrity after 6 hours, but the glomeruli and vasculatures were still recognizable up to 9 days in culture. We concluded that adult kidney tissue culture by hanging drop method can be used to study gene expressions in vasculature and glomeruli. © 2017 The Author(s). Published by S. Karger AG, Basel.

  2. Application of Hanging Drop Technique for Kidney Tissue Culture

    Directory of Open Access Journals (Sweden)

    Shaohui Wang

    2017-05-01

    Full Text Available Background/Aims: The hanging drop technique is a well-established method used in culture of animal tissues. However, this method has not been used in adult kidney tissue culture yet. This study was to explore the feasibility of using this technique for culturing adult kidney cortex to study the time course of RNA viability in the tubules and vasculature, as well as the tissue structural integrity. Methods: In each Petri dish with the plate covered with sterile buffer, a section of mouse renal cortex was cultured within a drop of DMEM culture medium on the inner surface of the lip facing downward. The tissue were then harvested at each specific time points for Real-time PCR analysis and histological studies. Results: The results showed that the mRNA level of most Na+ related transporters and cotransporters were stably maintained within 6 hours in culture, and that the mRNA level of most receptors found in the vasculature and glomeruli were stably maintained for up to 9 days in culture. Paraffin sections of the cultured renal cortex indicated that the tubules began to lose tubular integrity after 6 hours, but the glomeruli and vasculatures were still recognizable up to 9 days in culture. Conclusions: We concluded that adult kidney tissue culture by hanging drop method can be used to study gene expressions in vasculature and glomeruli.

  3. Nuclear techniques and in vitro culture for plant improvement

    International Nuclear Information System (INIS)

    1986-01-01

    The continuous series of food shortages in many parts of the world have led scientists to consider the possibilities of using the new techniques to develop better varieties of plants. The basis for plant breeding is suitable genetic variability and mutation induction as the means to create additional variation. In vitro techniques are a relatively new tool in practical plant breeding. These Proceedings contain 62 papers and posters presented at the symposium, as well as excerpts from the discussions. The Symposium presentations are divided into the following sessions: Genetic variation from in vitro culture; Genetic stability of in vitro cultures; In vitro culture with application of mutagens; Haploids; In vitro mutant selection; Use of genetic variation derived by in vitro culture; In vitro techniques as aids in mutation breeding and Genetic engineering. A separate abstract is prepared for each of these papers and posters

  4. Interspecific transfer of only part of genome by fusion between non-irradiated protoplasts of Nicotiana glauca and X-ray irradiated protoplasts of N. Langsdorffii

    International Nuclear Information System (INIS)

    Itoh, K.; Futsuhara, Y.

    1983-01-01

    To transfer only part of genome, X-ray irradiated suspension cell protoplasts of N. langsdorffii were fused with suspension cell protoplasts of N. glauca by polyethylene glycol. Somatic hybrid calli were selected by the growth in the hormone-free medium. Some of somatic hybrid calli from fusion with irradiated protoplasts indicated the loss of small subunit polypeptide of fraction 1 protein which was coded by N. langsdorffii nuclear DNA. Cytological analysis provided an information on significant decrease of chromosomes in somatic hybrid calli from fusion with irradiated protoplasts, compared with the somatic hybrid calli from fusion with non-irradiated protoplasts. In addition, isozyme analysis revealed that somatic hybrid calli from fusion with irradiated protoplasts lost particular bands of N. langsdorffli. These results demonstrate the tranfer of only part of genome from N, langsdorffii to N, glauca by fusion with X-ray irradiated protoplasts

  5. Cation selectivity of the plasma membrane of tobacco protoplasts in the electroporated state.

    Science.gov (United States)

    Wegner, Lars H

    2013-08-01

    Cation selectivity of the cellular membrane of tobacco culture cells (cell line 'bright yellow-2') exposed to pulsed electric fields in the millisecond range was investigated. The whole cell configuration of the patch clamp technique was established on protoplasts prepared from these cells. Ion selectivity of the electroporated membrane was investigated by measuring the reversal potential of currents passing through field-induced pores. To this end the membrane was hyper- or depolarized for 10ms (prepulse); subsequently the voltage was driven to opposite polarity at a constant rate (+40 or -40mV/ms, respectively). The experiment was started by polarizing the membrane to moderately negative or positive voltages (prepulse potential ±150mV) that would not induce pore formation. Subsequently, an extended voltage range was scanned in the porated state of the membrane (prepulse potential ±600mV). IV curves in the porated and the non-porated state (obtained at the same prepulse polarity) were superimposed to determine the voltage at which both curves intersected ('Intersection potential'). Using a modified version of the Goldmann-Hodgkin-Katz equation relative permeabilities to Ca(2+) and various monovalent alkali and organic cations were calculated. Pores were found to be fairly cation selective, with a selectivity sequence determined to be Ca(2+)>Li(+)>Rb(+)≈K(+)≈Na(+)>TEA(+)≈TBA(+)>Cl(-). Relative permeability to monovalent cations was inversely related to the ionic diameter. By fitting a formalism suggested by Dwyer at al. (J. Gen. Physiol. 75 (1980), 469-492) the effective average diameter of field induced pores was estimated to be about 1.8nm. Implications of these results for biotechnology and electroporation theory are discussed. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. Induction of recessive mutations in potato using tissue culture techniques

    International Nuclear Information System (INIS)

    Enckevort, L.J.G. van; Hoogkamp, T.J.H.; Bergervoet, J.E.M.; Visser, R.G.F.; Jacobsen, E.; Stiekma, W.J.; Pereira, A.

    2001-01-01

    In potato, two different in vitro approaches were used to generate recessive mutants. In the first method, monoploid plant material was irradiated to isolate and identify amylose-free (amf) mutants in potato. For isolating secondary mutants in the amf background new monoploids of the amf type were developed. A few selected amf monoploids showed excellent vigour in vitro, large leave; and microtuber formation. A diploid and a monoploid were tested for in vitro mutation induction and irradiated with 0 to 16 Gy X rays. The optimal dose for survival and mutation induction was between 4 and 8 Gy and plants were regenerated from irradiated leaf explants. In the second approach, mutants were induced by insertion of transposable elements in the diploids. This method was used to mutate R genes for resistance to Phytophthora infestans. Diploid heterozygous Rr plants with the immobilised Ds element, closely linked to one of the R genes, were selected. Mobilisation of Ds using Ac element transposase resulted in the selection of plants with active somatic Ds excision frequency of about 10%. In vitro protoplast isolation and plant regeneration from such plants enabled the selection of regenerants with new independent Ds insertions. Hygromycin selection (Ds excision marker on the T-DNA) during protoplast regeneration increased the frequency of Ds excision regenerants to 56%. A total of 582 hygromycin resistant plants were regenerated and selected in vitro. Preliminary analysis of the regenerants showed re-insertions of Ds in the predicted coding sequences of genes. (author)

  7. Tissue culture as a plant production technique for horticultural crops

    African Journals Online (AJOL)

    STORAGESEVER

    2009-08-18

    Aug 18, 2009 ... Recovery of regenerants from transformed cells. - Cell culture .... methods. Micropropagation techniques. Micropropagation is a simple concept. The basic pro- tocols were well established by the 1960s and a whole research field and ... the environment are naturally contaminated on their sur- faces (and ...

  8. Combined neutron imaging techniques for cultural heritage purpose

    International Nuclear Information System (INIS)

    Materna, T.

    2009-01-01

    This article presents the different new neutron techniques developed by the Ancient Charm collaboration to image objects of cultural heritage importance: Prompt-gamma-ray activation imaging (PGAI) coupled to cold/thermal neutron transmission tomography, Neutron Resonance Capture Imaging (NRCI) and Neutron Resonance Tomography.

  9. Propagation of Aquilaria malaccensis seedlings through tissue culture techniques

    International Nuclear Information System (INIS)

    Salahbiah Abdul Majid; Zaiton Ahmad; Mohd Rafaie Abdul Salam; Nurhayati Irwan; Affrida Abu Hassan; Rusli Ibrahim

    2010-01-01

    Aquilaria malaccensis or karas is the principal source of gaharu resin, which is used in many cultures for incense, perfumes and traditional medicines. The species is mainly propagated conventionally through seeds, cuttings and graftings. Propagation by seeds is usually a reliable method for other forest species, but for karas, this technique is inadequate to meet the current demand of seedling supplies. This is principally due to its low seed viability, low germination rate, delayed rooting of seedlings, long life-cycle and rare seed production. Tissue culture has several advantages over conventional propagation, especially for obtaining large number of uniform and high-yielding plantlets or clones. This paper presents the current progress on mass-propagation of Aquilaria malaccensis seedlings through tissue culture technique at Nuclear Malaysia. (author)

  10. Transformation of undomesticated strains of Bacillus subtilis by protoplast electroporation

    NARCIS (Netherlands)

    Romero, Diego; Perez-Garcia, Alejandro; Veening, Jan-Willem; de Vicente, Antonio; Kuipers, Oscar P.; de, Vicente A.

    A rapid method combining the use of protoplasts and electroporation was developed to transform recalcitrant wild strains of Bacillus subtilis. The method described here allows transformation with both replicative and integrative plasmids, as well as with chromosomal DNA, and provides a valuable tool

  11. Internalisation of cell-penetrating peptides into tobacco protoplasts.

    Science.gov (United States)

    Mäe, Maarja; Myrberg, Helena; Jiang, Yang; Paves, Heiti; Valkna, Andres; Langel, Ulo

    2005-05-20

    Cells are protected from the surrounding environment by plasma membrane which is impenetrable for most hydrophilic molecules. In the last 10 years cell-penetrating peptides (CPPs) have been discovered and developed. CPPs enter mammalian cells and carry cargo molecules over the plasma membrane with a molecular weight several times their own. Known transformation methods for plant cells have relatively low efficiency and require improvement. The possibility to use CPPs as potential delivery vectors for internalisation in plant cells has been studied in the present work. We analyse and compare the uptake of the fluorescein-labeled CPPs, transportan, TP10, penetratin and pVEC in Bowes human melanoma cells and Nicotiana tabacum cultivar (cv.) SR-1 protoplasts (plant cells without cell wall). We study the internalisation efficiency of CPPs with fluorescence microscopy, spectrofluorometry and fluorescence-activated cell sorter (FACS). All methods indicate, for the first time, that these CPPs can internalise into N. tabacum cv. SR-1 protoplasts. Transportan has the highest uptake efficacy among the studied peptides, both in mammalian cells and plant protoplast. The internalisation of CPPs by plant protoplasts may open up a new effective method for transfection in plants.

  12. Uniformity of plants regenerated from orange (Citrus sinensis Osb.) protoplasts.

    Science.gov (United States)

    Kobayashi, S

    1987-05-01

    Using 25 plants (protoclones) regenerated from orange (Citrus sinensis Osb.) protoplasts, several characters, including leaf and flower morphology, leaf oil, isozyme patterns and chromosome number, were examined. No significant variations in each character were recorded among the protoclones. Uniformity observed among protoclones was identical to that of nucellar seedlings.

  13. Genetic transformation of the white-rot fungus Dichomitus squalens using a new commercial protoplasting cocktail.

    Science.gov (United States)

    Daly, Paul; Slaghek, Gillian G; Casado López, Sara; Wiebenga, Ad; Hilden, Kristiina S; de Vries, Ronald P; Mäkelä, Miia R

    2017-12-01

    D. squalens, a white-rot fungus that efficiently degrades lignocellulose in nature, can be used in various biotechnological applications and has several strains with sequenced and annotated genomes. Here we present a method for the transformation of this basidiomycete fungus, using a recently introduced commercial ascomycete protoplasting enzyme cocktail, Protoplast F. In protoplasting of D. squalens mycelia, Protoplast F outperformed two other cocktails while releasing similar amounts of protoplasts to a third cocktail. The protoplasts released using Protoplast F had a regeneration rate of 12.5% (±6 SE). Using Protoplast F, the D. squalens monokaryon CBS464.89 was conferred with resistance to the antibiotics hygromycin and G418 via polyethylene glycol mediated protoplast transformation with resistance cassettes expressing the hygromycin phosphotransferase (hph) and neomycin phosphotransferase (nptII) genes, respectively. The hph gene was expressed in D. squalens using heterologous promoters from genes encoding β-tubulin or glyceraldehyde 3-phosphate dehydrogenase. A Southern blot confirmed integration of a resistance cassette into the D. squalens genome. An average of six transformants (±2 SE) were obtained when at least several million protoplasts were used (a transformation efficiency of 0.8 (±0.3 SE) transformants per μg DNA). Transformation of D. squalens demonstrates the suitability of the Protoplast F cocktail for basidiomycete transformation and furthermore can facilitate understanding of basidiomycete gene function and development of improved strains for biotechnological applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Plant regeneration from leaf protoplasts of Solanum torvum.

    Science.gov (United States)

    Guri, A; Volokita, M; Sink, K C

    1987-07-01

    A protocol to obtain regenerated plants from protoplasts of Solanum torvum Sw a wild species of eggplant resistant to Verticillium wilt is reported. Leaf protoplasts were enzymatically isolated from six-week old seedlings grown in a controlled environment chamber. Protoplasts were plated on modified KM medium (0.4 M glucose)+(mg/l): 1.0 p-chlorophenoxyacetic acid (CPA)+1.0 naphthaleneacetic acid (NAA)+0.5 6-benzylaminopurine (BAP) and 0.02 abscisic acid (ABA). The protoplast density was 5×10(4) per ml with 5 ml placed in each of two quadrants in X-dishes (100×15 mm). The reservoir medium was modified KM+(mg/l): 0.1 NAA+0.5 BAP+0.1 M sucrose+0.1 M mannitol+0.6% washed agar+1% activated charcoal. Dishes were initially placed in the dark at 27°C. Protoplast division was initiated in 1-2 weeks and 4 weeks later p-calli were 1-3 mm. Plating efficiency was 11% when measured at 3 weeks. Six-week old p-calli were transferred individually onto Whatman No. 1 filter paper layered on modified KM (0.15 M sucrose)+mg/l: 2.0 indoleacetic acid (IAA)+2.0 zeatin+0.5% washed agar for 2 weeks. Subsequently, shoots occurred within 4 weeks at 70% efficiency on MS+30 g/l sucrose+2 mg/l zeatin. Shoots were rooted on half strength MS+10 g/l sucrose.

  15. Protoplast isolation from Ulmus americana l. Pollen mother cells, tetrads, and microspores

    Energy Technology Data Exchange (ETDEWEB)

    Redenbaugh, M K; Westfall, R D; Karnosky, D F

    1980-01-01

    Meiotic protoplasts of U. amerciana are potentially valuable for producing interspecific elm hybrids through protoplast fusion. Meiotic cells(pollen mother cells, tetrads, and microspores) were incubated in either a cellulase, hemicellylase and pectinase enzyme solution of a beta-1,3-glucanase (lainarinase) solution. Respective protoplast isolation frequencies for the three meiotic cell types were 100, 50, and 10%. Exclusion staining with 0.2% Evans blue and 0.1% methyl blue suggested protoplast viability. Some of the microspore protoplasts were vacuolated, which is an important condition for cell division. Although attempts of regenerating cell walls and inducing cell division were unsuccessful, these two problems may be superceded by protoplast fusion with more regenerative protoplasts.

  16. Expression and epigenetic profile of protoplast cultures (Cucumis sativus L.)

    Czech Academy of Sciences Publication Activity Database

    Cápal, Petr; Ondřej, V.

    2014-01-01

    Roč. 50, č. 6 (2014), s. 789-794 ISSN 1054-5476 Institutional support: RVO:61389030 Keywords : SOMATIC EMBRYOGENESIS * DNA METHYLATION * CHROMATIN Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 0.981, year: 2014

  17. Antibodies to the CFTR modulate the turgor pressure of guard cell protoplasts via slow anion channels.

    Science.gov (United States)

    Leonhardt, N; Bazin, I; Richaud, P; Marin, E; Vavasseur, A; Forestier, C

    2001-04-06

    The plasma membrane guard cell slow anion channel is a key element at the basis of water loss control in plants allowing prolonged osmolite efflux necessary for stomatal closure. This channel has been extensively studied by electrophysiological approaches but its molecular identification is still lacking. Recently, we described that this channel was sharing some similarities with the mammalian ATP-binding cassette protein, cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel [Leonhardt, N. et al. (1999) Plant Cell 11, 1141-1151]. Here, using the patch-clamp technique and a bioassay, consisting in the observation of the change in guard cell protoplasts volume, we demonstrated that a functional antibody raised against the mammalian CFTR prevented ABA-induced guard cell protoplasts shrinking and partially inhibited the slow anion current. Moreover, this antibody immunoprecipitated a polypeptide from guard cell protein extracts and immunolabeled stomata in Vicia faba leaf sections. These results indicate that the guard cell slow anion channel is, or is closely controlled by a polypeptide, exhibiting one epitope shared with the mammalian CFTR.

  18. Somatic embryogenesis and plantlet regeneration from protoplast ...

    African Journals Online (AJOL)

    Administrator

    2010-05-30

    May 30, 2010 ... supplemented with 1 mg/l each of NAA and BA, 100 mg/l ascorbic acid and 0.5 M mannitol at 25°C in ... Key words: Ca-alginate beads, Muscari neglectum, nurse culture, plantlet regeneration, ..... Maddock SR (1987).

  19. Somatic embryogenesis and plantlet regeneration from protoplast ...

    African Journals Online (AJOL)

    After 4 weeks of culture, microcalli appeared on the surface of the Ca-alginate beads. Growth of microcalli in the medium with nurse cells (33.3%) was much better than those in the medium without nurse cells (6.5%). Transferring beads onto MS medium supplemented with 0.1 mg/l BA increased the growth of embryogenic ...

  20. The use of tissue culture techniques to detect irradiated vegetables

    International Nuclear Information System (INIS)

    Al-Safadi, B.; Sharabi, N.E.; Nabulsi, I

    2001-01-01

    the ability of two tissue culture methods, callus and vegetable growth induction, to detect irradiated vegetables was evaluated. Potato tubers, carrot roots, garlic cloves and onion bulbs were subjected to various gamma radiation doses (0, 25, 100, 150, 250, 500, 750, and 1000 Gy). Irradiated vegetables were cultured in vitro and in vivo (pots). Gamma irradiation significantly reduced callus-forming ability especially in carrot and potato where no callus was observed in doses higher than 50 Gy. Length of shoots and roots growing from irradiated garlic and onion explants was considerably reduced starting from the 25 Gy dose. No roots were formed on garlic explants at any irradiation dose. Garlic leaves growing from irradiated explants were spotted with purple to brown spots. The intensity of these spots increased as gamma ray dosage increased. In the pot experiment, potato plant appeared in the control only. On the contrary, a complete sprouting of garlic and onion was seen in all irradiation treatments. It was not possible to distinguish between the various irradiation treatments and the control 3 days after planting in pots. The two in vitro techniques, tested in our study, may effectively be used to detect irradiated vegetables and estimate the range of doses used. The callus formation method is more useful for potato and carrot, since regeneration of shoots in vitro from these two plants takes along time, making this method unpractical. The other technique is very useful in the case of onion and garlic since it is rapid. The two techniques can be used with most of the vegetables that can be cultured in vitro. (Author)

  1. Analytical techniques applied to study cultural heritage objects

    International Nuclear Information System (INIS)

    Rizzutto, M.A.; Curado, J.F.; Bernardes, S.; Campos, P.H.O.V.; Kajiya, E.A.M.; Silva, T.F.; Rodrigues, C.L.; Moro, M.; Tabacniks, M.; Added, N.

    2015-01-01

    The scientific study of artistic and cultural heritage objects have been routinely performed in Europe and the United States for decades. In Brazil this research area is growing, mainly through the use of physical and chemical characterization methods. Since 2003 the Group of Applied Physics with Particle Accelerators of the Physics Institute of the University of Sao Paulo (GFAA-IF) has been working with various methodologies for material characterization and analysis of cultural objects. Initially using ion beam analysis performed with Particle Induced X-Ray Emission (PIXE), Rutherford Backscattering (RBS) and recently Ion Beam Induced Luminescence (IBIL), for the determination of the elements and chemical compounds in the surface layers. These techniques are widely used in the Laboratory of Materials Analysis with Ion Beams (LAMFI-USP). Recently, the GFAA expanded the studies to other possibilities of analysis enabled by imaging techniques that coupled with elemental and compositional characterization provide a better understanding on the materials and techniques used in the creative process in the manufacture of objects. The imaging analysis, mainly used to examine and document artistic and cultural heritage objects, are performed through images with visible light, infrared reflectography (IR), fluorescence with ultraviolet radiation (UV), tangential light and digital radiography. Expanding more the possibilities of analysis, new capabilities were added using portable equipment such as Energy Dispersive X-Ray Fluorescence (ED-XRF) and Raman Spectroscopy that can be used for analysis 'in situ' at the museums. The results of these analyzes are providing valuable information on the manufacturing process and have provided new information on objects of different University of Sao Paulo museums. Improving the arsenal of cultural heritage analysis it was recently constructed an 3D robotic stage for the precise positioning of samples in the external beam setup

  2. Analytical techniques applied to study cultural heritage objects

    Energy Technology Data Exchange (ETDEWEB)

    Rizzutto, M.A.; Curado, J.F.; Bernardes, S.; Campos, P.H.O.V.; Kajiya, E.A.M.; Silva, T.F.; Rodrigues, C.L.; Moro, M.; Tabacniks, M.; Added, N., E-mail: rizzutto@if.usp.br [Universidade de Sao Paulo (USP), SP (Brazil). Instituto de Fisica

    2015-07-01

    The scientific study of artistic and cultural heritage objects have been routinely performed in Europe and the United States for decades. In Brazil this research area is growing, mainly through the use of physical and chemical characterization methods. Since 2003 the Group of Applied Physics with Particle Accelerators of the Physics Institute of the University of Sao Paulo (GFAA-IF) has been working with various methodologies for material characterization and analysis of cultural objects. Initially using ion beam analysis performed with Particle Induced X-Ray Emission (PIXE), Rutherford Backscattering (RBS) and recently Ion Beam Induced Luminescence (IBIL), for the determination of the elements and chemical compounds in the surface layers. These techniques are widely used in the Laboratory of Materials Analysis with Ion Beams (LAMFI-USP). Recently, the GFAA expanded the studies to other possibilities of analysis enabled by imaging techniques that coupled with elemental and compositional characterization provide a better understanding on the materials and techniques used in the creative process in the manufacture of objects. The imaging analysis, mainly used to examine and document artistic and cultural heritage objects, are performed through images with visible light, infrared reflectography (IR), fluorescence with ultraviolet radiation (UV), tangential light and digital radiography. Expanding more the possibilities of analysis, new capabilities were added using portable equipment such as Energy Dispersive X-Ray Fluorescence (ED-XRF) and Raman Spectroscopy that can be used for analysis 'in situ' at the museums. The results of these analyzes are providing valuable information on the manufacturing process and have provided new information on objects of different University of Sao Paulo museums. Improving the arsenal of cultural heritage analysis it was recently constructed an 3D robotic stage for the precise positioning of samples in the external beam setup

  3. IP3 stimulates CA++ efflux from fusogenic carrot protoplasts

    International Nuclear Information System (INIS)

    Rincon, M.; Boss, W.F.

    1986-01-01

    Polyphosphoinositide breakdown plays an important role in signal transduction in animal cells (Berridge and Irvine, 1984, Nature, 312:315). Upon stimulation, phospholipase C hydrolyzes phosphatidylinositol 4,5-bisphosphate to inositol 1,4,5-trisphosphate (IP 3 ) and diacylglycerol both of which act as cellular second messengers. IP 3 mobilizes Ca ++ from internal stores, hence the cytosolic free Ca ++ concentration increases and those physiological activities regulated by Ca ++ are stimulated. To test if plant cells also responded to IP 3 , Ca ++ efflux studies were done with fusogenic carrot protoplasts released in EGTA. The protoplasts were preloaded with 45 Ca ++ placed in a Ca ++ -free medium, and efflux determined as 45 Ca ++ loss from the protoplasts. IP 3 (10-20μM) caused enhanced 45 Ca ++ efflux and the response was sustained for at least 15 min. In plants, as in animals, the observed IP 3 -enhanced 45 Ca ++ efflux suggested that IP 3 released Ca ++ from internal stores, and the increased free cytosolic Ca ++ activated Ca ++ pumping mechanisms which restored the Ca ++ concentration in the cytosol to the normal level

  4. Ion beam analysis and spectrometry techniques for Cultural Heritage studies

    International Nuclear Information System (INIS)

    Beck, L.

    2013-01-01

    The implementation of experimental techniques for the characterisation of Cultural heritage materials has to take into account some requirements. The complexity of these past materials requires the development of new techniques of examination and analysis, or the transfer of technologies developed for the study of advanced materials. In addition, due to precious aspect of artwork it is also necessary to use the non-destructive methods, respecting the integrity of objects. It is for this reason that the methods using radiations and/or particles play a important role in the scientific study of art history and archaeology since their discovery. X-ray and γ-ray spectrometry as well as ion beam analysis (IBA) are analytical tools at the service of Cultural heritage. This report mainly presents experimental developments for IBA: PIXE, RBS/EBS and NRA. These developments were applied to the study of archaeological composite materials: layered materials or mixtures composed of organic and non-organic phases. Three examples are shown: evolution of silvering techniques for the production of counterfeit coinage during the Roman Empire and in the 16. century, the characterization of composites or mixed mineral/organic compounds such as bone and paint. In these last two cases, the combination of techniques gave original results on the proportion of both phases: apatite/collagen in bone, pigment/binder in paintings. Another part of this report is then dedicated to the non-invasive/non-destructive characterization of prehistoric pigments, in situ, for rock art studies in caves and in the laboratory. Finally, the perspectives of this work are presented. (author) [fr

  5. Characterisation and preservation of cultural heritage artefacts using nuclear techniques

    International Nuclear Information System (INIS)

    2011-01-01

    This report covers the studies performed for the identification and preservation of cultural heritage using nuclear analytical techniques (NAT). Within the context of the project financed by the IAEA, cultural articles from various excavation regions and from the Anatolian Civilizations Museum were analyzed and identified using the instruments at our Center and information was provided regarding their manufacturing techniques, past restoration history and socioeconomic indicators about the period within which these articles were used. The analysis of the articles which could not be removed from the museum were performed in-situ using portable instruments and support was provided to the experts for some articles from excavation regions for the evaluation of their originality. Within the framework the of this Project, five experts attended to the workshops and meetings organised by the IAEA and in the context of scientific visits and bilateral cooperation, one expert from Greece and three experts from Macedonia visited our Center and Anatolian Civilizations Museum and experimental studies were performed together

  6. From gene manipulation to forest establishment: shoot cultures of woody plants can be a central tool

    Energy Technology Data Exchange (ETDEWEB)

    McCown, B.H.

    1985-05-01

    Establishing germplasm of woody plants in microculture as shoot cultures has proved to be an effective method of overcoming many of the obstacles in working with these crops. Shoot cultures eliminate the changes associated with seasonal growth cycles and phase change and put large plants into a more manageable form. Well-established shoot cultures are central to successful clonal propagation systems for forest trees as well as to genetic improvement based on the use of cellular techniques such as protoplast manipulation. The physiological basis as to why tissues from shoot cultures are so readily manipulated is not well understood.

  7. The influence of organizational culture on the use of quality techniques and its impact on performance

    DEFF Research Database (Denmark)

    Gambi, Lillian; Jørgensen, Frances; Boer, Harry

    2013-01-01

    This report presents the results of a study about the influence of organizational culture on quality techniques and the impact of matching culture and technique to enhance performance. Data were drawn from 250 manufacturing companies in Brazil and Denmark. Profiles were identified according....... 2- Quality techniques contribute to improve performance, provided they are supported by appropriate cultural characteristics. For instance, the use of goal setting, continuous improvement, and failure prevention and control techniques supported by group and developmental cultures contribute...

  8. Characterisation of prototype Nurmi cultures using culture-based microbiological techniques and PCR-DGGE.

    Science.gov (United States)

    Waters, Sinéad M; Murphy, Richard A; Power, Ronan F G

    2006-08-01

    Undefined Nurmi-type cultures (NTCs) have been used successfully to prevent salmonella colonisation in poultry for decades. Such cultures are derived from the caecal contents of specific-pathogen-free birds and are administered via drinking water or spray application onto eggs in the hatchery. These cultures consist of many non-culturable and obligately anaerobic bacteria. Due to their undefined nature it is difficult to obtain approval from regulatory agencies to use these preparations as direct fed microbials for poultry. In this study, 10 batches of prototype NTCs were produced using an identical protocol over a period of 2 years. Traditional microbiological techniques and a molecular culture-independent methodology, polymerase chain reaction combined with denaturing gradient gel electrophoresis (PCR-DGGE), were applied to characterise these cultures and also to examine if the constituents of the NTCs were identical. Culture-dependent analysis of these cultures included plating on a variety of selective and semi-selective agars, examination of colony morphology, Gram-staining and a series of biochemical tests (API, BioMerieux, France). Two sets of PCR-DGGE studies were performed. These involved the amplification of universal and subsequently lactic acid bacteria (LAB)-specific hypervariable regions of a 16S rRNA gene by PCR. Resultant amplicons were subjected to DGGE. Sequence analysis was performed on subsequent bands present in resultant DGGE profiles using the Basic Local Alignment Search Tool (BLAST). Microbiological culturing techniques tended to isolate common probiotic bacterial species from the genera Lactobacillus, Lactococcus, Bifidobacterium, Enterococcus, Clostridium, Escherichia, Pediococcus and Enterobacterium as well as members of the genera, Actinomyces, Bacteroides, Propionibacterium, Capnocytophaga, Proteus, and Klebsiella. Bacteroides, Enterococcus, Escherichia, Brevibacterium, Klebsiella, Lactobacillus, Clostridium, Bacillus, Eubacterium

  9. Analysis of Cultural Heritage by Accelerator Techniques and Analytical Imaging

    Science.gov (United States)

    Ide-Ektessabi, Ari; Toque, Jay Arre; Murayama, Yusuke

    2011-12-01

    In this paper we present the result of experimental investigation using two very important accelerator techniques: (1) synchrotron radiation XRF and XAFS; and (2) accelerator mass spectrometry and multispectral analytical imaging for the investigation of cultural heritage. We also want to introduce a complementary approach to the investigation of artworks which is noninvasive and nondestructive that can be applied in situ. Four major projects will be discussed to illustrate the potential applications of these accelerator and analytical imaging techniques: (1) investigation of Mongolian Textile (Genghis Khan and Kublai Khan Period) using XRF, AMS and electron microscopy; (2) XRF studies of pigments collected from Korean Buddhist paintings; (3) creating a database of elemental composition and spectral reflectance of more than 1000 Japanese pigments which have been used for traditional Japanese paintings; and (4) visible light-near infrared spectroscopy and multispectral imaging of degraded malachite and azurite. The XRF measurements of the Japanese and Korean pigments could be used to complement the results of pigment identification by analytical imaging through spectral reflectance reconstruction. On the other hand, analysis of the Mongolian textiles revealed that they were produced between 12th and 13th century. Elemental analysis of the samples showed that they contained traces of gold, copper, iron and titanium. Based on the age and trace elements in the samples, it was concluded that the textiles were produced during the height of power of the Mongol empire, which makes them a valuable cultural heritage. Finally, the analysis of the degraded and discolored malachite and azurite demonstrates how multispectral analytical imaging could be used to complement the results of high energy-based techniques.

  10. Influences of explant type and enzyme incubation on isolated protoplast density and viability in two garlic cultivars

    International Nuclear Information System (INIS)

    Metwally, E.I.

    2014-01-01

    The present study reports on optimizing protoplast isolation and fusion in two garlic cultivars Balady and Seds 40. Protoplast density and viability were investigated in four different explants (etiolated and green parts of the pseudostem and lower and upper parts of the leaves) under enzyme incubation for 1, 2, 3 and 4 h. Among different explants, used for protoplast isolation in Balady cultivar, the upper and lower parts of the leaves produced the highest number of total protoplasts (70 and 66 pps/0.1 ml) at 4 and 3 h enzyme incubation, respectively. However, the etiolated part of pseudostem produced the highest number of viable protoplast in which 52.5 pps/0.1 ml were obtained at 3 h enzyme incubation. For protoplast isolation in Seds 40 cultivar, the highest number of total protoplasts (125 and 107.5 pps/0.1 ml) as well as viable protoplasts (105 and 107.5 pps/0.1 ml) was obtained from the etiolated and the green parts of pseudostem, respectively. The cultivar Seds 40 yielded higher total and viable protoplasts than Balady cultivar. Isolated protoplasts of Seds 40 and Balady were fused successfully at a protoplast density of 1 * 105 using either physical and/or electrical method. Optimization of the source of plant material as well as protoplast isolation conditions for garlic is a crucial step towards a successful protoplast fusion and subsequent colony formation. (author)

  11. Protection of the cultural environment by nuclear techniques

    International Nuclear Information System (INIS)

    Ramiere, R.

    1982-01-01

    The use of gamma radiation in the preservation of the cultural environment, and in particular art objects, is examined. In each branch, the present status of research on, and its application to, the treatment of art objects is described. Two major applications - disinfection and consolidation - are reviewed: (1) Disinfection: The destruction by gamma irradiation of xylophagous insects and of fungi which contaminate paintings, pictorial documents and mummies, is discussed. The effect of gamma-rays on the constituent materials of the infected objects is examined. (2) Consolidation: this process represents an extrapolation to the preservation of cultural objects of research performed over the last 20 years with the aim of improving the mechanical properties of porous materials (by impregnation of the material in a vacuum by a liquid plastic substance which is polymerized by radiation in situ). This technique is currently being used to treat ''uncoated'' wooden objects and, to a more limited extent, ''coated'' wood (polychromy and gilt wood), depending on the fragility of the covering. With regard to stone materials, the enhancement of the mechanical properties of a limestone impregnated with a styrene-polyester resin is described; this resin has been used to treat several sculptures. The use of acrylics and methacrylics has caused some samples to crack during radiation polymerization; theories put forward to explain this phenomenon are discussed. The consolidation of terracotta and bone has also been studied. The consolidation of waterlogged wood (archaeological wood) requires a different impregnation technique (diffusion in liquid). This preservation method has already been used to treat archaeological artifacts. (author)

  12. Tissue culture as a plant production technique for horticultural crops ...

    African Journals Online (AJOL)

    Over 100 years ago, Haberlandt envisioned the concept of plant tissue culture and provided the groundwork for the cultivation of plant cells, tissues and organs in culture. Initially plant tissue cultures arose as a research tool and focused on attempts to culture and study the development of small, isolated cells and segments ...

  13. Infection of cowpea protoplasts with sonchus yellow net virus and festuca leaf streak virus

    NARCIS (Netherlands)

    Beek, van N.A.M.

    1986-01-01

    The advantages of protoplast systems for plant virus research have been frequently reviewed (Zaitlin & Beachy, 1974; Takebe, 1975; Muhlbach, 1982; Sander & Mertens, 1984). Relatively little attention has been given to the limitations of such a system.

    Protoplasts do not

  14. A simple and effective method to encapsulate tobacco mesophyll protoplasts to maintain cell viability

    Directory of Open Access Journals (Sweden)

    Rong Lei

    2015-01-01

    • It is very convenient to change or collect the solution without mechanically disturbing the protoplasts. This simple and effective silica sol–gel/alginate two-step immobilization of protoplasts in Transwell has great potential for applications in genetic transformation, metabolite production, and migration assays.

  15. Application of optical tweezers and excimer laser to study protoplast fusion

    Science.gov (United States)

    Kantawang, Titirat; Samipak, Sompid; Limtrakul, Jumras; Chattham, Nattaporn

    2015-07-01

    Protoplast fusion is a physical phenomenon that two protoplasts come in contact and fuse together. Doing so, it is possible to combine specific genes from one protoplast to another during fusion such as drought resistance and disease resistance. There are a few possible methods to induce protoplast fusion, for example, electrofusion and chemical fusion. In this study, chemical fusion was performed with laser applied as an external force to enhance rate of fusion and observed under a microscope. Optical tweezers (1064 nm with 100X objective N.A. 1.3) and excimer laser (308 nm LMU-40X-UVB objective) were set with a Nikon Ti-U inverted microscope. Samples were prepared by soaking in hypertonic solution in order to induce cell plasmolysis. Elodea Canadensis and Allium cepa plasmolysed leaves were cut and observed under microscope. Concentration of solution was varied to induce difference turgor pressures on protoplasts pushing at cell wall. Free protoplasts in solution were trapped by optical tweezers to study the effect of Polyethylene glycol (PEG) solution. PEG was diluted by Ca+ solution during the process to induced protoplast cell contact and fusion. Possibility of protoplast fusion by excimer laser was investigated and found possible. Here we report a novel tool for plant cell fusion using excimer laser. Plant growth after cell fusion is currently conducted.

  16. Formation and regeneration of protoplasts and spheroplasts of gastrointestinal strains of lactobacilli.

    OpenAIRE

    Connell, H; Lemmon, J; Tannock, G W

    1988-01-01

    Methods were developed for the formation of protoplasts and spheroplasts of gastrointestinal strains of Lactobacillus reuteri, Lactobacillus gasseri, and Lactobacillus salivarius. Attempts to regenerate vegetative cells from protoplasts were not successful, but spheroplasts could be regenerated consistently for five of six strains.

  17. Do phosphoinositides regulate membrane water permeability of tobacco protoplasts by enhancing the aquaporin pathway?

    Science.gov (United States)

    Ma, Xiaohong; Shatil-Cohen, Arava; Ben-Dor, Shifra; Wigoda, Noa; Perera, Imara Y; Im, Yang Ju; Diminshtein, Sofia; Yu, Ling; Boss, Wendy F; Moshelion, Menachem; Moran, Nava

    2015-03-01

    Enhancing the membrane content of PtdInsP 2 , the already-recognized protein-regulating lipid, increased the osmotic water permeability of tobacco protoplasts, apparently by increasing the abundance of active aquaporins in their membranes. While phosphoinositides are implicated in cell volume changes and are known to regulate some ion channels, their modulation of aquaporins activity has not yet been reported for any organism. To examine this, we compared the osmotic water permeability (P f) of protoplasts isolated from tobacco (Nicotiana tabacum) cultured cells (NT1) with different (genetically lowered or elevated relative to controls) levels of inositol trisphosphate (InsP3) and phosphatidyl inositol [4,5] bisphosphate (PtdInsP2). To achieve this, the cells were transformed with, respectively, the human InsP3 5-phosphatase ('Ptase cells') or human phosphatidylinositol (4) phosphate 5-kinase ('PIPK cells'). The mean P f of the PIPK cells was several-fold higher relative to that of controls and Ptase cells. Three results favor aquaporins over the membrane matrix as underlying this excessive P f: (1) transient expression of the maize aquaporin ZmPIP2;4 in the PIPK cells increased P f by 12-30 μm s(-1), while in the controls only by 3-4 μm s(-1). (2) Cytosol acidification-known to inhibit aquaporins-lowered the P f in the PIPK cells down to control levels. (3) The transcript of at least one aquaporin was elevated in the PIPK cells. Together, the three results demonstrate the differences between the PIPK cells and their controls, and suggest a hitherto unobserved regulation of aquaporins by phosphoinositides, which could occur through direct interaction or indirect phosphoinositides-dependent cellular effects.

  18. [Isolation and regeneration of the protoplasts of the streptomycete producers of actinomycins C and X].

    Science.gov (United States)

    Orlova, T I; Masha, G G; Kliueva, N A

    1986-09-01

    Protoplasts of S. michiganensis, S. chrysomallus and Streptomyces sp. 26-115, organisms producing actinomycins C and X form in hypertonic salt solution under the action of 3-4,5 mg/ml of lysozyme on the mycelium suspension. For protoplasting, the streptomycetes were grown on the soybean medium in the presence of 0.2-0.8 per cent of glycine. The mycelium of the streptomycete exponential growth phase was more favourable for protoplast formation. Protoplast regeneration was studied on the medium described by Okanishi et al. The quantitative composition of this medium was not optimal for regeneration of protoplasts of the above streptomycetes. The level of their regeneration depended to various extents on concentration of phosphate, magnesium and calcium ions and sucrose in the regeneration medium.

  19. Liposome-enhanced transformation of Streptococcus lactis and plasmid transfer by intergeneric protoplast fusion of Streptococcus lactis and Bacillus subtilis

    NARCIS (Netherlands)

    Vossen, Jos M.B.M. van der; Kok, Jan; Lelie, Daniel van der; Venema, Gerhardus

    An efficient protoplast transformation system and a procedure of plasmid transfer by means of protoplast fusion is described for Streptococcus lactis. Protoplasts of S. lactis IL1403 and S. lactis MG1363 were transformed by pGK12 [2.9 MDa erythromycin resistance (Emr)] with an efficiency of 3 × 10^5

  20. In vitro techniques for crop improvement

    International Nuclear Information System (INIS)

    1985-01-01

    The film refers to principles of plant tissue culture - laboratory requirements, media preparation, explant establishment and subculturing method. In vitro growth and development of crop plants are demonstrate and the application of in vitro techniques in plant breeding is discussed. The second part of the film shows the application of cell, tissue and organ culture in plants. Micropropagation and virus eradication are important technologies for the improvement of vegetatively propagated plants; zygotic embryo rescue techniques are used for distant hybridization, especially in cereals. Plant biotechnology offers a potent means for the in vitro generation of enhanced genetic variability - somaclonal and mutagen induced variation. Principles of the isolation and culture of plant protoplasts are explained and their potential for somatic hybridization in higher plants is demonstrated. Haploids are valuable to accelerate breeding cycles of plants. Plant biotechnology is described as an important complementary tool to conventional plant breeding methods

  1. Efficient Transformation of Oil Palm Protoplasts by PEG-Mediated Transfection and DNA Microinjection

    Science.gov (United States)

    Masani, Mat Yunus Abdul; Noll, Gundula A.; Parveez, Ghulam Kadir Ahmad; Sambanthamurthi, Ravigadevi; Prüfer, Dirk

    2014-01-01

    Background Genetic engineering remains a major challenge in oil palm (Elaeis guineensis) because particle bombardment and Agrobacterium-mediated transformation are laborious and/or inefficient in this species, often producing chimeric plants and escapes. Protoplasts are beneficial as a starting material for genetic engineering because they are totipotent, and chimeras are avoided by regenerating transgenic plants from single cells. Novel approaches for the transformation of oil palm protoplasts could therefore offer a new and efficient strategy for the development of transgenic oil palm plants. Methodology/Principal Findings We recently achieved the regeneration of healthy and fertile oil palms from protoplasts. Therefore, we focused on the development of a reliable PEG-mediated transformation protocol for oil palm protoplasts by establishing and validating optimal heat shock conditions, concentrations of DNA, PEG and magnesium chloride, and the transfection procedure. We also investigated the transformation of oil palm protoplasts by DNA microinjection and successfully regenerated transgenic microcalli expressing green fluorescent protein as a visible marker to determine the efficiency of transformation. Conclusions/Significance We have established the first successful protocols for the transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection. These novel protocols allow the rapid and efficient generation of non-chimeric transgenic callus and represent a significant milestone in the use of protoplasts as a starting material for the development of genetically-engineered oil palm plants. PMID:24821306

  2. Teaching Organizational Culture Using a Projective Technique: Collage Construction

    Science.gov (United States)

    Colakoglu, Saba; Littlefield, Jon

    2011-01-01

    Although the topic of "organizational culture" is an integral part of syllabi across a wide range of core business classes such as Principles of Management, Organizational Behavior, and Human Resource Management, few experiential exercises exist that can enhance student understanding and learning of different layers of organizational culture. In…

  3. Analysis of culture-dependent versus culture-independent techniques for identification of bacteria in clinically obtained bronchoalveolar lavage fluid.

    Science.gov (United States)

    Dickson, Robert P; Erb-Downward, John R; Prescott, Hallie C; Martinez, Fernando J; Curtis, Jeffrey L; Lama, Vibha N; Huffnagle, Gary B

    2014-10-01

    The diagnosis and management of pneumonia are limited by the use of culture-based techniques of microbial identification, which may fail to identify unculturable, fastidious, and metabolically active viable but unculturable bacteria. Novel high-throughput culture-independent techniques hold promise but have not been systematically compared to conventional culture. We analyzed 46 clinically obtained bronchoalveolar lavage (BAL) fluid specimens from symptomatic and asymptomatic lung transplant recipients both by culture (using a clinical microbiology laboratory protocol) and by bacterial 16S rRNA gene pyrosequencing. Bacteria were identified in 44 of 46 (95.7%) BAL fluid specimens by culture-independent sequencing, significantly more than the number of specimens in which bacteria were detected (37 of 46, 80.4%, P ≤ 0.05) or "pathogen" species reported (18 of 46, 39.1%, P ≤ 0.0001) via culture. Identification of bacteria by culture was positively associated with culture-independent indices of infection (total bacterial DNA burden and low bacterial community diversity) (P ≤ 0.01). In BAL fluid specimens with no culture growth, the amount of bacterial DNA was greater than that in reagent and rinse controls, and communities were markedly dominated by select Gammaproteobacteria, notably Escherichia species and Pseudomonas fluorescens. Culture growth above the threshold of 10(4) CFU/ml was correlated with increased bacterial DNA burden (P Microbiology. All Rights Reserved.

  4. Tomato protoplast DNA transformation : physical linkage and recombination of exogenous DNA sequences

    NARCIS (Netherlands)

    Jongsma, Maarten; Koornneef, Maarten; Zabel, Pim; Hille, Jacques

    1987-01-01

    Tomato protoplasts have been transformed with plasmid DNA's, containing a chimeric kanamycin resistance gene and putative tomato origins of replication. A calcium phosphate-DNA mediated transformation procedure was employed in combination with either polyethylene glycol or polyvinyl alcohol. There

  5. A comparison of different Gracilariopsis lemaneiformis (Rhodophyta) parts in biochemical characteristics, protoplast formation and regeneration

    Science.gov (United States)

    Wang, Zhongxia; Sui, Zhenghong; Hu, Yiyi; Zhang, Si; Pan, Yulong; Ju, Hongri

    2014-08-01

    Gracilariopsis lemaneiformis is a commercially exploited alga. Its filaceous thallus can be divided into three parts, holdfast, middle segment and tip. The growth and branch forming trend and agar content of these three parts were analyzed, respectively, in this study. The results showed that the tip had the highest growth rate and branched most, although it was the last part with branch forming ability. The holdfast formed branches earliest but slowly. Holdfast had the highest agar content. We also assessed the difference in protoplast formation and regeneration among three parts. The middle segment displayed the shortest enzymolysis time and the highest protoplast yield; whereas the tip had the strongest vitality of protoplasts formation. Juvenile plants were only obtained from the protoplasts generated from the tip. These results suggested that the differentiation and function of G. lemaneiformis was different.

  6. Evaluation of conventional castaneda and lysis centrifugation blood culture techniques for diagnosis of human brucellosis.

    Science.gov (United States)

    Mantur, Basappa G; Mangalgi, Smita S

    2004-09-01

    We investigated the role of the lysis centrifugation blood culture technique over the conventional Castaneda technique for the diagnosis of human brucellosis. The lysis centrifugation technique has been found to be more sensitive in both acute (20% higher sensitivity; P centrifugation was in the mean detection time, which was only 2.4 days in acute and 2.7 days in chronic cases, with 103 out of 110 (93.6%) and 17 out of 20 (85%) cultures from acute and chronic brucellosis, respectively, detected before the conventional culture was positive. Our results confirmed the potential usefulness of the lysis technique in diagnosis and institution of appropriate antibiotic therapy.

  7. CATEGORICAL IMAGE COMPONENTS IN THE FORMING SYSTEM OF A MARKETING TECHNIQUES MANAGER’S IMAGE CULTURE

    OpenAIRE

    Anna Borisovna Cherednyakova

    2015-01-01

    Based on the understanding of the image culture formation of managers of marketing techniques, as a representative of the social and communication interaction of public structures, categorical apparatus of image culture with an emphasis on the etymology of the image, as an integral component of image culture was analyzed. Categorical components of the image are presented from the standpoint of image culture, as personal new formation, an integral part of the professional activity of the marke...

  8. Effect of Radiation Dosage on Efficiency of Chloroplast Transfer by Protoplast Fusion in Nicotiana

    OpenAIRE

    Menczel, László; Galiba, Gábor; Nagy, Ferenc; Maliga, Pál

    1982-01-01

    Chloroplasts of Nicotiana tabacum SR1 were transferred into Nicotiana plumbaginifolia by protoplast fusion. The protoplasts of the organelle donor were irradiated with different lethal doses using a 60Co source, to facilitate the elimination of their nuclei from the fusion products. After fusion induction, clones derived from fusion products and containing streptomycin-resistant N. tabacum SR1 chloroplasts were selected by their ability to green on a selective medium. When N. tabacum protopla...

  9. The use of flow cytometry to monitor chitin synthesis in regenerating protoplasts of Candida albicans.

    Science.gov (United States)

    Hector, R F; Braun, P C; Hart, J T; Kamarck, M E

    1990-01-01

    Flow cytometry was used to monitor chitin synthesis in regenerating protoplasts of the yeast Candida albicans. Comparisons of cells stained with Calcofluor White, a fluorochrome with known affinity for chitin, and cells incubated in the presence of N-[3H]-acetylglucosamine, the precursor substrate for chitin, showed a linear relationship between fluorescence and incorporation of label over time. Changes in both the fluorescence and light scatter of regenerating protoplasts treated with inhibitors of fungal chitin synthase were also quantitated by flow cytometry.

  10. A study on the isolation of protoplasts from mesophyll cells of Dendrobium Queen Pink

    International Nuclear Information System (INIS)

    Aqeel, R.; Zehra, M.; Kazmi, S. K.; Khan, S.

    2016-01-01

    Protoplasts were successfully isolated from one month old In vitro grown plantlets of Dendrobium cultivar Queen pink. The enzyme solution used was composed of 1 percent Cellulase Onozuka R-10, 0.5 percent Macerozyme R-10, 0.1 percent Pectinase, 0.3 M mannitol, 10 mM CaCl/sub 2/.2H/sub 2/O and 10 mM 2 (N-morpholino)-ethanesulfonic acid (MES) at pH 5.8. Protoplast highest yield with 15.7x104 protoplasts per 1.5 gm freshly chopped leaves were obtained when digested in enzyme solution for 4 hrs on a rotary shaker with an agitation speed of 45 rpm in dark conditions. Protoplasts were filtered with 45 micro m nylon sieve and washed with 0.3 M mannitol solution supplemented with 10 mM CaCl/sub 2/.2H/sub 2/O and 10 mM MES, and purified with 0.3 M sucrose solution gradient. Purification of protoplasts on a sucrose mannitol gradient yielded clean protoplasts that were free from debris. (author)

  11. 14C fixation by leaves and leaf cell protoplasts of the submerged aquatic angiosperm Potamogeton lucens: Carbon dioxide or bicarbonate?

    International Nuclear Information System (INIS)

    Staal, M.; Elzenga, J.T.M.; Prins, H.B.A.

    1989-01-01

    Protoplasts were isolated from leaves of the aquatic angiosperm Potamogeton lucens L. The leaves utilize bicarbonate as a carbon source for photosynthesis, and show polarity; that is acidification of the periplasmic space of the lower, and alkalinization of the space near the upper leaf side. At present there are two models under consideration for this photosynthetic bicarbonate utilization process: conversion of bicarbonate into free carbon dioxide as a result of acidification and, second, a bicarbonate-proton symport across the plasma membrane. Carbon fixation of protoplasts was studied at different pH values and compared with that in leaf strips. Using the isotopic disequilibrium technique, it was established that carbon dioxide and not bicarbonate was the form in which DIC actually crossed the plasma membrane. It is concluded that there is probably no true bicarbonate transport system at the plasma membrane of these cells and that bicarbonate utilization in this species apparently rests on the conversion of bicarbonate into carbon dioxide. Experiments with acetazolamide, an inhibitor of periplasmic carbonic anhydrase, and direct measurements of carbonic anhydrase activity in intact leaves indicate that in this species the role of this enzyme for periplasmic conversion of bicarbonate into carbon dioxide is insignificant

  12. Comparison of Nested-PCR technique and culture method in ...

    African Journals Online (AJOL)

    USER

    2010-04-05

    Apr 5, 2010 ... Full Length Research Paper. Comparison of ... The aim of the present study was to evaluate the diagnostic value of nested PCR in genitourinary ... method. Based on obtained results, the positivity rate of urine samples in this study was 5.0% by using culture and PCR methods and 2.5% for acid fast staining.

  13. From cultural aesthetic to perfor- mance technique: continuities and ...

    African Journals Online (AJOL)

    Introduction. A broad-based survey of Malawian dances that I undertook several years ago aimed rather ambitiously to identify persistent regional forms of these dances as well as their governing aesthetic(s). In the course of the field work, a recurrent ~estion emerged: how do we approach dance as cultural performance?

  14. Human mixed lymphocyte cultures. Evaluation of microculture technique utilizing the multiple automated sample harvester (MASH)

    Science.gov (United States)

    Thurman, G. B.; Strong, D. M.; Ahmed, A.; Green, S. S.; Sell, K. W.; Hartzman, R. J.; Bach, F. H.

    1973-01-01

    Use of lymphocyte cultures for in vitro studies such as pretransplant histocompatibility testing has established the need for standardization of this technique. A microculture technique has been developed that has facilitated the culturing of lymphocytes and increased the quantity of cultures feasible, while lowering the variation between replicate samples. Cultures were prepared for determination of tritiated thymidine incorporation using a Multiple Automated Sample Harvester (MASH). Using this system, the parameters that influence the in vitro responsiveness of human lymphocytes to allogeneic lymphocytes have been investigated. PMID:4271568

  15. CATEGORICAL IMAGE COMPONENTS IN THE FORMING SYSTEM OF A MARKETING TECHNIQUES MANAGER’S IMAGE CULTURE

    Directory of Open Access Journals (Sweden)

    Anna Borisovna Cherednyakova

    2015-08-01

    Full Text Available Based on the understanding of the image culture formation of managers of marketing techniques, as a representative of the social and communication interaction of public structures, categorical apparatus of image culture with an emphasis on the etymology of the image, as an integral component of image culture was analyzed. Categorical components of the image are presented from the standpoint of image culture, as personal new formation, an integral part of the professional activity of the marketing techniques manager: object-communicative categorical component, subject-activity categorical component of image, personality-oriented categorical component, value-acmeological categorical component of image.The aim is to identify and justify the image categorical components as a component of image culture of the marketing techniques manager.Method and methodology of work – a general scientific research approach reflecting scientific apparatus of research.Results. Categorical components of the image, as an image culture component of manager of marketing techniques were defined.Practical implication of the results. The theoretical part of «Imageology» course, special course «Image culture of manager of marketing techniques», the theoretical and methodological study and the formation of image culture.

  16. Citrus asymmetric somatic hybrids produced via fusion of gamma-irradiated and iodoacetamide-treated protoplasts

    Energy Technology Data Exchange (ETDEWEB)

    Bona, Claudine Maria de [Instituto Agronomico do Parana (IAPAR), Curitiba, PR (Brazil)], e-mail: debona@iapar.br; Gould, Jean Howe [Texas A and M University, College Station, TX (United States). Dept. of Ecosystem Science and Management], e-mail: gould@tamu.edu; Miller Junior, J. Creighton [Texas A and M University, College Station, TX (United States). Dept. of Horticultural Sciences], e-mail: jcmillerjr@tamu.edu; Stelly, David [Texas A and M University, College Station, TX (United States). Dept. of Soil and Crop Sciences], e-mail: stelly@tamu.edu; Louzada, Eliezer Silva [Texas A and M University, Kingsville, TX (United States). Citrus Center], e-mail: e-louzada@tamu.edu

    2009-05-15

    The objective of this study was to produce citrus somatic asymmetric hybrids by fusing gamma.irradiated protoplasts with iodoacetamide-treated protoplasts. Protoplasts were isolated from embryogenic suspension cells of grapefruit (Citrus paradisi Macfad.) cultivars Ruby Red and Flame, sweet oranges (C. sinensis Osbeck) 'Itaborai', 'Natal', Valencia', and 'Succari', from 'Satsuma' (C. unshiu Marcow.) and 'Changsha' mandarin (C. reticulata Blanco) and 'Murcott' tangor (C. reticulata x C. sinensis). Donor protoplasts were exposed to gamma rays and receptor protoplasts were treated with 3 mmol L{sup -1} iodoacetamide (IOA), and then they were fused for asymmetric hybridization. Asymmetric embryos were germinated, and the resulting shoots were either grafted onto sour orange, rough lemon or 'Swingle' (C. paradisi x Poncirus trifoliata) x 'Sunki' mandarin rootstock seedlings, or rooted after dipping their bases in indol.butyric acid (IBA) solution. The products were later acclimatized to greenhouse conditions. Ploidy was analyzed by flow cytometry, and hybridity was confirmed by amplified fragment length polymorphism (AFLP) analysis of plantlet DNA samples. The best treatment was the donor-recipient fusion combination of 80 Gy.irradiated 'Ruby Red' protoplasts with 20 min IOA.treated 'Succari' protoplasts. Tetraploid and aneuploid plants were produced. Rooting recalcitrance was solved by dipping shoots' stems in 3,000 mg L{sup -1} IBA solution for 10 min. (author)

  17. Effect of postmortem sampling technique on the clinical significance of autopsy blood cultures.

    Science.gov (United States)

    Hove, M; Pencil, S D

    1998-02-01

    Our objective was to investigate the value of postmortem autopsy blood cultures performed with an iodine-subclavian technique relative to the classical method of atrial heat searing and antemortem blood cultures. The study consisted of a prospective autopsy series with each case serving as its own control relative to subsequent testing, and a retrospective survey of patients coming to autopsy who had both autopsy blood cultures and premortem blood cultures. A busy academic autopsy service (600 cases per year) at University of Texas Medical Branch Hospitals, Galveston, Texas, served as the setting for this work. The incidence of non-clinically relevant (false-positive) culture results were compared using different methods for collecting blood samples in a prospective series of 38 adult autopsy specimens. One hundred eleven adult autopsy specimens in which both postmortem and antemortem blood cultures were obtained were studied retrospectively. For both studies, positive culture results were scored as either clinically relevant or false positives based on analysis of the autopsy findings and the clinical summary. The rate of false-positive culture results obtained by an iodine-subclavian technique from blood drawn soon after death were statistically significantly lower (13%) than using the classical method of obtaining blood through the atrium after heat searing at the time of the autopsy (34%) in the same set of autopsy subjects. When autopsy results were compared with subjects' antemortem blood culture results, there was no significant difference in the rate of non-clinically relevant culture results in a paired retrospective series of antemortem blood cultures and postmortem blood cultures using the iodine-subclavian postmortem method (11.7% v 13.5%). The results indicate that autopsy blood cultures obtained using the iodine-subclavian technique have reliability equivalent to that of antemortem blood cultures.

  18. Project 'Use of nuclear techniques in investigation, conservation and management of the cultural historical patrimony

    International Nuclear Information System (INIS)

    Kochmann, Sonnia

    2000-12-01

    This project is aimed at solving problems of conservation of the cultural historical patrimony through the active participation of the member countries of ARCAL by the application of Analytic Nuclear Techniques [es

  19. Micropropagation of Dalbergia sissoo Roxb. through tissue culture technique.

    Science.gov (United States)

    Sahu, Jyoti; Khan, Shagufta; Sahu, Ram Kumar; Roy, Amit

    2014-04-01

    Multiple shoots of Dalbergia sissoo Roxb. (Sissoo) were incited from seeds through indirect somatic embryogenesis method. Seeds were inoculated in Murashige and Skoog's medium without any growth hormone. Than cotyledonary leaves were struck and used for callus induction on MS medium amplified with 2, 4-dichlorophenoxyacetic acid (0.5 to 4 mg mL(-1)). After 3 to 4 weeks the embryogenic callus clumps was transferred to medium supplemented with cytokinin (BAP 1 to 5 mg L(-1), kinetin 1-5.0 mg L(-1)) for embryo maturation and germination. The high-frequency shoot proliferation (82%) and maximum number of shoots per explants were recorded in MS medium containing NAA (0.5)+BAP (0.5). The findings of recent investigations have shown that, it is possible to induce indirect somatic embryogenesis in Dalbergia sissoo and plant regeneration from callus cultures derived from cotyledonary leaves as explants.

  20. An optimized protocol for handling and processing fragile acini cultured with the hanging drop technique.

    Science.gov (United States)

    Snyman, Celia; Elliott, Edith

    2011-12-15

    The hanging drop three-dimensional culture technique allows cultivation of functional three-dimensional mammary constructs without exogenous extracellular matrix. The fragile acini are, however, difficult to preserve during processing steps for advanced microscopic investigation. We describe adaptations to the protocol for handling of hanging drop cultures to include investigation using confocal, scanning, and electron microscopy, with minimal loss of cell culture components. Copyright © 2011 Elsevier Inc. All rights reserved.

  1. Cultural Variations in Mothers' Acceptance of and Intent to Use Behavioral Child Management Techniques

    Science.gov (United States)

    Mah, Janet W. T.; Johnston, Charlotte

    2012-01-01

    We examined cultural differences in mothers' acceptance of and intent to use behavioral parenting techniques for managing disruptive child behavior, and the possible roles of parenting styles and implicit theories in explaining these cultural differences. A community sample of 117 Euro-Canadian and Chinese-immigrant mothers of boys aged 4- to…

  2. Analysis of DNA polymerase activity in Petunia protoplasts treated with clastogenic agents

    International Nuclear Information System (INIS)

    Benediktsson, I.; Spampinato, C.P.; Andreo, C.S.; Schieder, O.

    1994-01-01

    Clastogenic agents, i.e. agents that can induce chromosome or DNA breakage, have been shown to enhance the role of direct gene transfer to protoplasts. The effect was analysed at the enzymatic level using protoplast homogenates as well as intact protoplasts. For that purpose existing procedures were modified to enable measurement of DNA polymerase in vivo. In the system used, external DNA was able to enter the cells without the addition of membrane-permeabilizing compounds. When comparing total DNA polymerase activity of protoplasts irradiated with X-rays or UV-light with that of untreated cells we did not observe significant differences. Incubation of protoplasts with high doses of bleomycin affected total DNA polymerase activity negatively. but dideoxythymidine triphosphate-sensitive activity was not influenced. We conclude that the DNA strand-breaks induced by low doses of X-rays. UV-light or bleomycin do not increase the total or the repair-DNA polymerase activity and. therefore. that the increase in the transformation rates after DNA strand-breaking is not preceded by enhanced DNA polymerase activity. (author)

  3. Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory.

    Science.gov (United States)

    Bowey-Dellinger, Kristen; Dixon, Luke; Ackerman, Kristin; Vigueira, Cynthia; Suh, Yewseok K; Lyda, Todd; Sapp, Kelli; Grider, Michael; Crater, Dinene; Russell, Travis; Elias, Michael; Coffield, V McNeil; Segarra, Verónica A

    2017-01-01

    Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented-one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.

  4. The retraction of the protoplast during PCD is an active, and interruptible, calcium-flux driven process.

    Science.gov (United States)

    Kacprzyk, Joanna; Brogan, Niall P; Daly, Cara T; Doyle, Siamsa M; Diamond, Mark; Molony, Elizabeth M; McCabe, Paul F

    2017-07-01

    The protoplast retracts during apoptosis-like programmed cell death (AL-PCD) and, if this retraction is an active component of AL-PCD, it should be used as a defining feature for this type of programmed cell death. We used an array of pharmacological and genetic tools to test if the rates of protoplast retraction in cells undergoing AL-PCD can be modulated. Disturbing calcium flux signalling, ATP synthesis and mitochondrial permeability transition all inhibited protoplast retraction and often also the execution of the death programme. Protoplast retraction can precede loss of plasma membrane integrity and cell death can be interrupted after the protoplast retraction had already occurred. Blocking calcium influx inhibited the protoplast retraction, reduced DNA fragmentation and delayed death induced by AL-PCD associated stresses. At higher levels of stress, where cell death occurs without protoplast retraction, blocking calcium flux had no effect on the death process. The results therefore strongly suggest that retraction of the protoplast is an active biological process dependent on an early Ca 2+ -mediated trigger rather than cellular disintegration due to plasma membrane damage. Therefore this morphologically distinct cell type is a quantifiable feature, and consequently, reporter of AL-PCD. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Molecular characterization of intergeneric hybrid between Aspergillus oryzae and Trichoderma harzianum by protoplast fusion.

    Science.gov (United States)

    Patil, N S; Patil, S M; Govindwar, S P; Jadhav, J P

    2015-02-01

    Protoplast fusion between Aspergillus oryzae and Trichoderma harzianum and application of fusant in degradation of shellfish waste. The filamentous chitinolytic fungal strains A. oryzae NCIM 1272 and T. harzianum NCIM 1185 were selected as parents for protoplast fusion. Viable protoplasts were released from fungal mycelium using enzyme cocktail containing 5 mg ml(-1) lysing enzymes from T. harzianum, 0.06 mg ml(-1) β-glucuronidase from Helix pomatia and 1 mg ml(-1) purified Penicillium ochrochloron chitinase in 0.8 mol l(-1) sorbitol as an osmotic stabilizer. Intergeneric protoplast fusion was carried out using 60% polyethylene glycol as a fusogen. At optimum conditions, the regeneration frequency of the fused protoplasts on colloidal chitin medium and fusion frequency were calculated. Fusant showed higher rate of growth pattern, chitinase activity and protein content than parents. Fusant formation was confirmed by morphological markers, viz. colony morphology and spore size and denaturation gradient gel electrophoresis (DGGE). This study revealed protoplast fusion between A. oryzae and T. harzianum significantly enhanced chitinase activity which ultimately provides potential strain for degradation of shellfish waste. Consistency in the molecular characterization results using DGGE is the major outcome of this study which can be emerged as a fundamental step in fusant identification. Now it is need to provide attention over effective chitin degradation to manage shrimp processing issues. In this aspect, ability of fusant to degrade shellfish waste efficiently in short incubation time revealed discovery of potential strain in the reclamation of seafood processing crustacean bio-waste. © 2014 The Society for Applied Microbiology.

  6. Ability of Bacillus subtilis protoplasts to repair irradiated bacteriophage deoxyribonucleic acid via acquired and natural enzymatic systems

    International Nuclear Information System (INIS)

    Yasbin, R.E.; Andersen, B.J.; Sutherland, B.M.

    1981-01-01

    A novel form of enzyme therapy was achieved by utilizing protoplasts of Bacillus subtilis. Photoreactivating enzyme of Escherichia coli was successfully inserted into the protoplasts of B. subtilis treated with polyethylene glycol. This enzyme was used to photoreactivate ultraviolet-damaged bacteriophage deoxyribonucleic acid (DNA). Furthermore, in polyethylene glycol-treated protoplasts, ultraviolet-irradiated transfecting bacteriophage DNA was shown to be a functional substrate for the host DNA excision repair system. Previous results (R.E. Yasbin, J.D. Fernwalt, and P.I. Fields, J. Bacteriol.; 137: 391-396) showed that ultraviolet-irradiated bacteriophage DNA could not be repaired via the excision repair system of competent cells. Therefore, the processing of bacteriophage DNA by protoplasts and by competent cells must be different. This sensitive protoplast assay can be used to identify and to isolate various types of DNA repair enzymes

  7. Study on production of useful metabolites by development of advanced cell culture techniques using radiation

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Byung Yeoup; Kim, Jin Hong; Lee, Seung Sik; Kim, Jae Sung; An, Byung Chull; Moon, Yu Ran; Lee, Eun Mi; Lee, Min Hee; Lee, Jae Tack [KAERI, Daejeon (Korea, Republic of)

    2010-02-15

    The purpose of this project is improvement of investigation, materialization and evaluation techniques on effectiveness for functional natural compounds throughout development of tissue/cell culture techniques for mass production of useful metabolites using radiation. Research scope includes 1) Development of a technique for radiation tissue and cell culture, 2) Database construction for radiation response in plants and radiation effects, 3) Construction of general-purpose national based techniques of cell culture technique using radiation. Main results are as follow: mass culture of the adventitious roots of mountain ginseng (Panax ginseng C. A. Meyer) roots using rare earth elements in bioreactor: characterization of a transcription factor EoP gene from centipedegrass and the transcription regulation of LexA from Synechocystis sp PCC6803 and E. coli: identification of gamma-ray induced hydrogenase synthesis in hox gene transformed E. coli: transformation and the selection of the EoP transgene from Arabidopsis, rice and lettuce: Identification of the maysin and maysin derivatives in centipedegrass: characterization of gamma-ray induced color change in Taxus cuspidata: verification of the expression of antioxidant proteins (POD, APX and CAT) to gamma-ray in Arabidopsis: comparison of the response of the expression level to gamma-ray or H{sub 2}O{sub 2} in Arabidopsis; verification of the responses and effects to gamma-ray from plants (analysis of NPQ and ROS levels): the development method for rapidly enhancing maysin content of centipede grass; establishment of mass culture system for red beet

  8. Study on Production of Useful Metabolites by Development of Advanced Cell Culture Techniques Using Radiation

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Byung Yeoup; Kim, J. H.; Lee, S. S.; Shyamkumar, B.; An, B. C.; Moon, Y. R.; Lee, E. M.; Lee, M. H.

    2009-02-15

    The purpose of this project is improvement of investigation, materialization and evaluation techniques on effectiveness for functional natural compounds throughout development of tissue/cell culture techniques for mass production of useful metabolites using radiation. Research scope includes 1) Development of a technique for radiation tissue and cell culture, 2) Database construction for radiation response in plants and radiation effects, 3) Construction of general-purpose national based techniques of cell culture technique using radiation. Main results are as follow: Establishment of a tissue culture system (Rubus sp., Lithospermum erythrorhizon, and Rhodiola rosea); characterization of radiation activated gene expression from cultivated bokbunja (Rubus sp.) and Synechocystis sp., identification of gamma-ray induced color change in plants; identification of sensitivity to gamma-ray from Omija (Schisandra chinensis) extract; identification of the response of thylakoid proteins to gamma-ray in spinach and Arabidopsis; identification of gamma-ray induced gene relating to pigment metabolism; characterization of different NPQ changes to gamma-irradiated plants; verification of the effects of rare earth element including anti-bacterial and anti-fungal properties and as a growth enhancer; identification of changes in the growth of gamma-irradiated Synechocystis; and investigation of liquid cell culture conditions from Rhodiola rosea

  9. Study on production of useful metabolites by development of advanced cell culture techniques using radiation

    International Nuclear Information System (INIS)

    Chung, Byung Yeoup; Kim, Jin Hong; Lee, Seung Sik; Kim, Jae Sung; An, Byung Chull; Moon, Yu Ran; Lee, Eun Mi; Lee, Min Hee; Lee, Jae Tack

    2010-02-01

    The purpose of this project is improvement of investigation, materialization and evaluation techniques on effectiveness for functional natural compounds throughout development of tissue/cell culture techniques for mass production of useful metabolites using radiation. Research scope includes 1) Development of a technique for radiation tissue and cell culture, 2) Database construction for radiation response in plants and radiation effects, 3) Construction of general-purpose national based techniques of cell culture technique using radiation. Main results are as follow: mass culture of the adventitious roots of mountain ginseng (Panax ginseng C. A. Meyer) roots using rare earth elements in bioreactor: characterization of a transcription factor EoP gene from centipedegrass and the transcription regulation of LexA from Synechocystis sp PCC6803 and E. coli: identification of gamma-ray induced hydrogenase synthesis in hox gene transformed E. coli: transformation and the selection of the EoP transgene from Arabidopsis, rice and lettuce: Identification of the maysin and maysin derivatives in centipedegrass: characterization of gamma-ray induced color change in Taxus cuspidata: verification of the expression of antioxidant proteins (POD, APX and CAT) to gamma-ray in Arabidopsis: comparison of the response of the expression level to gamma-ray or H 2 O 2 in Arabidopsis; verification of the responses and effects to gamma-ray from plants (analysis of NPQ and ROS levels): the development method for rapidly enhancing maysin content of centipede grass; establishment of mass culture system for red beet

  10. Study on Production of Useful Metabolites by Development of Advanced Cell Culture Techniques Using Radiation

    International Nuclear Information System (INIS)

    Chung, Byung Yeoup; Kim, J. H.; Lee, S. S.; Shyamkumar, B.; An, B. C.; Moon, Y. R.; Lee, E. M.; Lee, M. H.

    2009-02-01

    The purpose of this project is improvement of investigation, materialization and evaluation techniques on effectiveness for functional natural compounds throughout development of tissue/cell culture techniques for mass production of useful metabolites using radiation. Research scope includes 1) Development of a technique for radiation tissue and cell culture, 2) Database construction for radiation response in plants and radiation effects, 3) Construction of general-purpose national based techniques of cell culture technique using radiation. Main results are as follow: Establishment of a tissue culture system (Rubus sp., Lithospermum erythrorhizon, and Rhodiola rosea); characterization of radiation activated gene expression from cultivated bokbunja (Rubus sp.) and Synechocystis sp., identification of gamma-ray induced color change in plants; identification of sensitivity to gamma-ray from Omija (Schisandra chinensis) extract; identification of the response of thylakoid proteins to gamma-ray in spinach and Arabidopsis; identification of gamma-ray induced gene relating to pigment metabolism; characterization of different NPQ changes to gamma-irradiated plants; verification of the effects of rare earth element including anti-bacterial and anti-fungal properties and as a growth enhancer; identification of changes in the growth of gamma-irradiated Synechocystis; and investigation of liquid cell culture conditions from Rhodiola rosea

  11. Study on production of useful metabolites by development of advanced cell culture techniques using radiation

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Byung Yeoup; Lee, Seung Sik; Bai, Hyounwoo; Singh, Sudhir; Lee, Eun Mi; Hong, Sung Hyun; Park, Chul Hong; Srilatha, B.; Kim, Mi Ja; Lee, Ohchul

    2012-01-15

    The purpose of this project is improvement of investigation, materialization and evaluation techniques on effectiveness for functional natural compounds throughout development of tissue/cell culture techniques for mass production of useful metabolites using radiation. Research scope includes Development of a technique for radiation tissue and cell culture, Database construction for radiation response in plants and radiation effects, Construction of general-purpose national based techniques of cell culture technique using radiation. Main results are as follow: Development of a technique for radiation tissue and cell culture for Erigeron breviscapus (Vant.) Hand. Mazz.; Identification and functional analysis of AtTDX (chaperone and peroxidase activities); Functional analysis of radiation(gamma ray, electron beam, and proton beam) induced chaperon protein activities (AtTDX); Determine the action mechanism of yPrx2; Development of transgenic plant with bas I gene from Arabidopsis; Development of transgenic plant with EoP gene from centipedegrass; Identification of radiation induced multi functional compounds from Aloe; Determination of the effects of radiation on removing undesirable color and physiological activities (Schizandra chinensis baillon, centipedegrass); Determine the action mechanism of transgenic plant with 2-Cys Prx for heat stress resistance; Determination of the effects of centipedegrass extracts on anti-cancer activities; Functional analysis of centipedegrass extracts (anti-virus effects)

  12. Protoplast formation, regeneration and transformation from the taxol ...

    African Journals Online (AJOL)

    producing fungus Ozonium sp. BT2 were investigated, including the enzymolysis time and temperature, the osmotic pressure stabilizer, mycelial incubation time, the culture medium, the culture methods and preprocessing. The mycelia were ...

  13. PEDAGIGOCAL TECHNIQUE OF BUILDING THE CULTURE OF INTERPERSONAL RELATIONS IN PRESCHOOL CHILDREN AT ART CLASSES

    Directory of Open Access Journals (Sweden)

    Svetlana Vyacheslavovna Kahnovich

    2013-10-01

    Full Text Available The article looks at the pedagogical technique of building the culture of interpersonal relations in preschool children at the local and modular level. Interpersonal relations are viewed as the module and art classes as the local level. The research is timely as it can assist in studying the problem of moral development of preschool children by building the culture of interpersonal relations by artistic education means. The study presents novelty concluding from the survey of scientific literature. The process of building the culture of interpersonal relations in children has not been properly studied by preschool pedagogy. The task of the present study is to elaborate a pedagogical technique to build the culture of interpersonal relations between children at art classes. The article discusses ‘technological’ criteria (term by G.K. Selevko and presents interactive principles of the pedagogical technique. Group activities alongside with individual ones were viewed as organizational forms of art classes. Building the culture of interpersonal relations in preschool children at art classes is closely connected with the development of their personality, a child’s  consciousness, their motivational and conceptual spheres during their gradual moral development at various levels - emotional (attitude, axiological level, psychic (intentional cognitive processes, activity (artistic and interpersonal literacy. Graphic (projective methods were used to analyze age dynamics of ethical and moral development. The conclusion describes a set of pedagogical conditions for efficient building of the culture of interpersonal relations in children at art classes.  Goal. To elaborate a pedagogical technique for building the culture of interpersonal relations in preschool children at art classes. The technique can be applied at local and modular level.Methods and Methodology. The pedagogical technique is aimed at building the culture of interpersonal relations

  14. Improvement of rice anther culture and application of the technique in mutation breeding

    International Nuclear Information System (INIS)

    Chen Qiufang; Wang Cailian; Lu Yimei; Jin Wei

    2001-01-01

    The ability of callus formation and green plant regeneration was very different for different rice type and varieties in anther culture. The differentiation and regeneration of green plants were obviously improved when the rice anthers at about 30 d after culture on induction medium were irradiated with 20 Gy of γ-rays and calli were cultured on the differentiation medium containing 30 mg/L colchicines. The stimulation effect of γ-irradiation combined with colchicines was much better than that of their single use. Mutation frequency and selective efficiency in M 2 were obviously increased by application of the technique

  15. Protoplast isolation, transient transformation of leaf mesophyll protoplasts and improved Agrobacterium-mediated leaf disc infiltration of Phaseolus vulgaris: tools for rapid gene expression analysis.

    Science.gov (United States)

    Nanjareddy, Kalpana; Arthikala, Manoj-Kumar; Blanco, Lourdes; Arellano, Elizabeth S; Lara, Miguel

    2016-06-24

    Phaseolus vulgaris is one of the most extensively studied model legumes in the world. The P. vulgaris genome sequence is available; therefore, the need for an efficient and rapid transformation system is more imperative than ever. The functional characterization of P. vulgaris genes is impeded chiefly due to the non-amenable nature of Phaseolus sp. to stable genetic transformation. Transient transformation systems are convenient and versatile alternatives for rapid gene functional characterization studies. Hence, the present work focuses on standardizing methodologies for protoplast isolation from multiple tissues and transient transformation protocols for rapid gene expression analysis in the recalcitrant grain legume P. vulgaris. Herein, we provide methodologies for the high-throughput isolation of leaf mesophyll-, flower petal-, hypocotyl-, root- and nodule-derived protoplasts from P. vulgaris. The highly efficient polyethylene glycol-mannitol magnesium (PEG-MMG)-mediated transformation of leaf mesophyll protoplasts was optimized using a GUS reporter gene. We used the P. vulgaris SNF1-related protein kinase 1 (PvSnRK1) gene as proof of concept to demonstrate rapid gene functional analysis. An RT-qPCR analysis of protoplasts that had been transformed with PvSnRK1-RNAi and PvSnRK1-OE vectors showed the significant downregulation and ectopic constitutive expression (overexpression), respectively, of the PvSnRK1 transcript. We also demonstrated an improved transient transformation approach, sonication-assisted Agrobacterium-mediated transformation (SAAT), for the leaf disc infiltration of P. vulgaris. Interestingly, this method resulted in a 90 % transformation efficiency and transformed 60-85 % of the cells in a given area of the leaf surface. The constitutive expression of YFP further confirmed the amenability of the system to gene functional characterization studies. We present simple and efficient methodologies for protoplast isolation from multiple P

  16. Improvement of polysaccharide and triterpenoid production of Ganoderma lucidum through mutagenesis of protoplasts

    Directory of Open Access Journals (Sweden)

    Ren Peng

    2016-03-01

    Full Text Available Ganoderma lucidum is a traditional medicinal macrofungus in China, which has two kinds of key bioactive compounds -- polysaccharides and triterpenoids. To improve the polysaccharide and triterpenoid production from G. lucidum, the preparation and regeneration conditions of protoplasts were optimized. This was done by systematic trials with various parameters, and protoplast mutation was subsequently performed. A mycelium that was cultivated for seven days and treated with 0.33 mL of 1% snailase and 0.66 mL of 0.5% cellulase solution for 2.5 h at 30 °C in the presence of osmotic pressure stabilizer mannitol (0.5 mol/L, had the best conditions, in which the resultant protoplasts were 6.40 × 105/mL and the regeneration rate was 6.25%. The resultant protoplasts were subjected to subsequent mutation by lithium chloride or by the combination of lithium chloride and Triton X-100. The highest yields of intracellular polysaccharide and triterpenoid in two mutant strains were 37.50 and 40.81 mg/g, which were increased with 568.45% and 373.43%, respectively, as compared to the original strain. Furthermore, the yields of intracellular polysaccharides and triterpenoids in the second generation and the third generation of the mutants were comparable to that of the first generation, which showed genetic stability of the mutants for the production of polysaccharides and triterpenoids.

  17. Induction and catabolite repression of α-glucosidase synthesis in protoplasts of Saccharomyces carlsbergensis

    NARCIS (Netherlands)

    Wijk, R. van; Ouwehand, J.; Bos, T. van den; Koningsberger, V.V.

    1969-01-01

    1. 1. Kinetic data on the repression, the derepression and the induction of α-glucosidase synthesis in protoplasts of Saccharomyces carlsbergensis suggested that some site other than the stereospecific site for the induction by maltose was involved in the repression by glucose. 2. 2. A study of the

  18. AGGREGATION AND FUSION OF PLANT-PROTOPLASTS AFTER SURFACE-LABELING WITH BIOTIN AND AVIDIN

    NARCIS (Netherlands)

    VANKESTEREN, WJP; MOLEMA, E; TEMPELAAR, MJ

    1993-01-01

    In mass electrofusion systems with aggregation of protoplasts by alignment, the yield and composition of fusion products can be predicted by a simple model. Through computer simulation, upper limits were found for the yield of binary and multi fusions. To overcome constraints on binary products,

  19. Study on production of useful metabolites by development of advanced cell culture techniques using radiation

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Byung Yeoup; Kim, Jinhong; Lee, Seung Sik; Bai, Hyounwoo; An, Byung Chull; Lee, Eun Mi; Lee, Jae Taek; Kim, Mi Ja

    2010-12-15

    The purpose of this project is improvement of investigation, materialization and evaluation techniques on effectiveness for functional natural compounds throughout development of tissue/cell culture techniques for mass production of useful metabolites using radiation. Research scope includes Development of a technique for radiation tissue and cell culture, Database construction for radiation response in plants and radiation effects, Construction of general-purpose national based techniques of cell culture technique using radiation. Main results are as follow: Isolation and identification of radiation induced basI gene; Determination of stresses sensitivities by transformating basI gene into arabidopsis; Isolation and identification of radiation induced chaperon proteins (PaAhpC and yPrxII) from Pseudomonas and yeast, and structural and functional analysis of the proteins; Determination of oxidative and heat resistance by transformating PaAhpC; Isolation and identification of maysin and its derivatives from centipedgrass; Investigation of enhancement technique for improving maysin and its derivatives production using radiation; Investigation of removing undesirable color in maysin and its derivatives using radiation; Determination of the effect of radiation on physiological functions of centipedgrass extracts; Identification of H{sub 2}O{sub 2} removing enzyme in radiation irradiated plant (Spinach); Determination of the effects of centipedgrass extracts on anti-obesity and anti-cancer activities.

  20. Study on production of useful metabolites by development of advanced cell culture techniques using radiation

    International Nuclear Information System (INIS)

    Chung, Byung Yeoup; Kim, Jinhong; Lee, Seung Sik; Bai, Hyounwoo; An, Byung Chull; Lee, Eun Mi; Lee, Jae Taek; Kim, Mi Ja

    2010-12-01

    The purpose of this project is improvement of investigation, materialization and evaluation techniques on effectiveness for functional natural compounds throughout development of tissue/cell culture techniques for mass production of useful metabolites using radiation. Research scope includes Development of a technique for radiation tissue and cell culture, Database construction for radiation response in plants and radiation effects, Construction of general-purpose national based techniques of cell culture technique using radiation. Main results are as follow: Isolation and identification of radiation induced basI gene; Determination of stresses sensitivities by transformating basI gene into arabidopsis; Isolation and identification of radiation induced chaperon proteins (PaAhpC and yPrxII) from Pseudomonas and yeast, and structural and functional analysis of the proteins; Determination of oxidative and heat resistance by transformating PaAhpC; Isolation and identification of maysin and its derivatives from centipedgrass; Investigation of enhancement technique for improving maysin and its derivatives production using radiation; Investigation of removing undesirable color in maysin and its derivatives using radiation; Determination of the effect of radiation on physiological functions of centipedgrass extracts; Identification of H 2 O 2 removing enzyme in radiation irradiated plant (Spinach); Determination of the effects of centipedgrass extracts on anti-obesity and anti-cancer activities

  1. Protoplasting impact on polyketide activity and characterization of the interspecific fusants from Streptomyces spp

    International Nuclear Information System (INIS)

    Slama, N.; Lazim, H.; Barkallah, Insaf; Abbassi, M.; Ben Hassen, A.; Limam, F.

    2009-01-01

    Streptomycetes are gram-positive, soil-inhabiting bacteria of the order Actinomycetales. These organisms exhibit an unusual, developmentally complex life cycle and produce many economically important secondary metabolites, such as antibiotics, immunosuppressants, insecticides, and antitumor agents. Streptomyces species have been the subject of genetic investigation for over 50 years, with many studies focusing on the production of bioactives compounds. The protoplast formation and regeneration are important processes, and they are a major step following genetic manipulations such as fusion and DNA-mediated transformation, which can improve antibiotic production. The protoplast fusion, transformation and improved fermentation features can be used to regenerate strains with increased antibiotic activity. Local Streptomyces spp. CN207 produce a broad range of secondary metabolites which is active against bacteria and fungi. This strain was used as a donor and S. coelicolor strain M145 was used as a recipient host for protoplast fusion. The protoplast fusion resulted in increased isolation of variants with higher antibiotic activity. Recombinant Streptomyces coelicolor PF04 was increased 10 times more than the wild strain. The antimicrobial activity from PF04 strain was studied using the disc method agar. TLC analysis confirmed that the Rf of cell extract for PF04 strain is identical to antimicrobial compound of Streptomyces CN207. Our results confirm the possibility of transferring antibiotics cluster genes by fusion. In fact, many of the selective markers such as Ticarcillin, Cefalotin, Oxacillin and Cefotaxim were transferred during the protoplast fusion. PFGE analysis and DNA-hybridization confirmed the presence of homologous fragments between a wild-type Streptomyces CN207 and a recombinant S. coelicolor PF04

  2. Technique of the 'in vitro' fertilization and the culture of mouse embryos at preimplantation

    International Nuclear Information System (INIS)

    Kikuchi, Olivia Kimiko; Yamada, Takeshi

    1993-03-01

    The mammal embryo is an intensive cellular proliferating system, very radiosensitive and therefore adequate to the study of the biological effects of ionizing radiation. The technique of the in vitro fertilization and the culture of mouse embryos at preimplantation period, modified by Yamada et al (1982) to improve the efficiency of more than 95% of blastocyst formation is described. (author)

  3. Agricultural production - Phase 2. Indonesia. Rice - azolla - fish culture - use of nuclear technique

    International Nuclear Information System (INIS)

    Watanabe, Iwao.

    1991-01-01

    The primary aim of the expert mission was to provide advice on the use of nuclear techniques to study rice-azolla-fish culture. Results of the work performed so far show that basal application of azolla gives similar or better yields of rice than basal application of urea. Fish productivity was also found to be significantly higher when azolla is present. 2 tabs

  4. In vitro culture techniques as a tool of sugarcane bud germination ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-10-20

    Oct 20, 2008 ... and shoot growth under salt stress with different NaCl concentrations (0, 17, 34, 68 and 102 mM) using cultivar NCo310. ... techniques could be used to evaluate salt stress effects in sugarcane at the germination stage. Key words: in vitro culture, .... l'Enseignement Supérieur, de la Formation des Cadres et.

  5. A thin-layer liquid culture technique for the growth of Helicobacter pylori.

    Science.gov (United States)

    Joo, Jung-Soo; Park, Kyung-Chul; Song, Jae-Young; Kim, Dong-Hyun; Lee, Kyung-Ja; Kwon, Young-Cheol; Kim, Jung-Min; Kim, Kyung-Mi; Youn, Hee-Shang; Kang, Hyung-Lyun; Baik, Seung-Chul; Lee, Woo-Kon; Cho, Myung-Je; Rhee, Kwang-Ho

    2010-08-01

    Several attempts have been successful in liquid cultivation of Helicobaccter pylori. However, there is a need to improve the growth of H. pylori in liquid media in order to get affluent growth and a simple approach for examining bacterial properties. We introduce here a thin-layer liquid culture technique for the growth of H. pylori. A thin-layer liquid culture system was established by adding liquid media to a 90-mm diameter Petri dish. Optimal conditions for bacterial growth were investigated and then viability, growth curve, and released proteins were examined. Maximal growth of H. pylori was obtained by adding 3 mL of brucella broth supplemented with 10% horse to a Petri dish. H. pylori grew in both DMEM and RPMI-1640 supplemented with 10% fetal bovine serum and 0.5% yeast extract. Serum-free RPMI-1640 supported the growth of H. pylori when supplemented with dimethyl-beta-cyclodextrin (200 microg/mL) and 1% yeast extract. Under optimal growth, H. pylori grew exponentially for 28 hours, reaching a density of 3.4 OD(600) with a generation time of 3.3 hours. After 24 hours, cultures at a cell density of 1.0 OD(600) contained 1.3 +/- 0.1 x 10(9 )CFU/mL. gamma-Glutamyl transpeptidase, nuclease, superoxide dismutase, and urease were not detected in culture supernatants at 24 hours in thin-layer liquid culture, but were present at 48 hours, whereas alcohol dehydrogenase, alkylhydroperoxide reductase, catalase, and vacuolating cytotoxin were detected at 24 hours. Thin-layer liquid culture technique is feasible, and can serve as a versatile liquid culture technique for investigating bacterial properties of H. pylori.

  6. Protoplast formation, regeneration and transformation from the taxol ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-06-17

    Jun 17, 2008 ... BT2 were investigated, including the enzymolysis time and temperature, the osmotic pressure stabilizer, mycelial incubation time, the culture medium, the culture methods and preprocessing. The mycelia were ... foundation of fungi genetic manipulation and improvement but also a good experimental ...

  7. Single Spore Isolation as a Simple and Efficient Technique to obtain fungal pure culture

    Science.gov (United States)

    Noman, E.; Al-Gheethi, AA; Rahman, N. K.; Talip, B.; Mohamed, R.; H, N.; Kadir, O. A.

    2018-04-01

    The successful identification of fungi by phenotypic methods or molecular technique depends mainly on the using an advanced technique for purifying the isolates. The most efficient is the single spore technique due to the simple requirements and the efficiency in preventing the contamination by yeast, mites or bacteria. The method described in the present work is depends on the using of a light microscope to transfer one spore into a new culture medium. The present work describes a simple and efficient procedure for single spore isolation to purify of fungi recovered from the clinical wastes.

  8. Ex Vivo Produced Oral Mucosa Equivalent by Using the Direct Explant Cell Culture Technique

    Directory of Open Access Journals (Sweden)

    Kamile Öztürk

    2012-09-01

    Full Text Available Objective: The aim of this study is the histological and immunohistochemical evaluation of ex vivo produced oral mucosal equivalents using keratinocytes cultured by direct explant technique.Material and Methods: Oral mucosa tissue samples were obtained from the keratinized gingival tissues of 14 healthy human subjects. Human oral mucosa keratinocytes from an oral mucosa biopsy specimen were dissociated by the explant technique. Once a sufficient population of keratinocytes was reached, they were seeded onto the type IV collagen coated “AlloDerm” and taken for histological and immunohistochemical examinations at 11 days postseeding of the keratinocytes on the cadaveric human dermal matrix.Results: Histopathologically and immunohistochemically, 12 out of 14 successful ex vivo produced oral mucosa equivalents (EVPOME that consisted of a stratified epidermis on a dermal matrix have been developed with keratinocytes cultured by the explant technique.Conclusion: The technical handling involved in the direct explant method at the beginning of the process has fewer steps than the enzymatic method and use of the direct explant technique protocol for culturing of human oral mucosa keratinocyte may be more adequate for EVPOME production.

  9. Measurement techniques and safety culture in radiation protection -reflections after 37 years of occupation with measuring instruments

    International Nuclear Information System (INIS)

    Maushart, R.

    1994-01-01

    Safety Culture in radiation use and radiation protection implies primarily knowledge and competence of the decision makers. As the measuring techniques are basic for practical radiation protection, only such person can be called competent who has sufficient expertise on measuring techniques, and is able to evaluate its application and results. Safety Culture also implies the readiness to expose errors, and to learn from them. ''Believing in infallibility'' excludes Safety Culture. Therefore, correctly applied measuring technique contributes to recognize weak points early. How far it is used consciously and actively to prevent undesirable developments and exceeding of limits, can be considered outright as a yardstick for a high-ranking safety culture. Safety Culture as a whole, however, needs more than more measuring techniques. It requires its own and adequate Measurement Culture, presupposing also motivation and determination to measure. Therefore, education, training, knowledge and consciousness of safety of the people who are responsible for measurements are decisive for successful radiation protection. (orig.) [de

  10. Examination, characterisation and analysis techniques for the knowledge and the conservation / restoration of cultural heritage - importance of ionising radiation techniques

    International Nuclear Information System (INIS)

    Boutaine; J. L.

    2004-01-01

    For the examination, characterisation and analysis of cultural heritage artefacts or art objects and their component materials, the conservation scientist needs a palette of non destructive and non invasive techniques, in order to improve our knowledge concerning their elaboration, their evolution and/or degradation during time, and to give rational basis for their restoration and conservation. A general survey and illustrations showing the usefulness of these techniques will be presented. Among these methods, many are based on the use of ionising radiation. 1. Radiography (using X-rays, gamma rays, beta particles, secondary electrons, neutrons), electron emission radiography, tomodensimetry, 2. Scanning electron microscope associated with X-ray spectrometry, 3. X-ray diffraction, 4. Synchrotron radiation characterisation, 5. X-ray fluorescence analysis, 6. Activation analysis, 7. Ion beam analysis (PIXE, PIGE, RBS, secondary X-ray fluorescence), 8. Thermoluminescence dating, 9. Carbon-14 dating. These methods are used alone or in connection with other analytical methods. Any kind of materials can be encountered, for instance: i. stones, gems, ceramics, terracotta, enamels, glasses, i i. wood, paper, textile, bone, ivory, i i i. metals, jewellery, i v. paint layers, canvas and wooden backings, pigments, dyers, oils, binding media, varnishes, glues. Some examples will be taken, among recent work done at the Centre of Research and Restoration of the Museums of France (C2RMF), from various geographical origins, various ages and different art disciplines. This will illustrate the kind of assistance that science and technology can provide to a better knowledge of mankind's cultural heritage and also to the establishment of rational basis for its better conservation for the future generations. (Author)

  11. La3+ uptake and its effect on the cytoskeleton in root protoplasts of Zea mays L.

    Science.gov (United States)

    Liu, Min; Hasenstein, Karl H

    2005-03-01

    La(3+) ions are known to antagonize Ca(2+) and are used as a Ca(2+) channel blocker but little is known on the direct effects of La(3+). Micromolar La(3+) concentrations promoted root growth while higher concentrations were inhibitory. The uptake of La(3+) in maize root protoplasts revealed a membrane binding component (0.14 and 0.44 pmol min(-1) protoplast(-1) for 100 and 1,000 microM La(3+)) followed by a slower concentration and time-dependent uptake. Uptake was reduced by Ca(2+), but had no substantial effect on other ions. La(3+) shifted microtubule organization from random to parallel but caused aggregation of microfilaments. Our data suggest that La(3+) is taken up into plant cells and affects growth via stabilization of the cytoskeleton.

  12. Improved inhibitor tolerance in xylose-fermenting yeast Spathaspora passalidarum by mutagenesis and protoplast fusion

    DEFF Research Database (Denmark)

    Hou, Xiaoru; Yao, Shuo

    2012-01-01

    The xylose-fermenting yeast Spathaspora passalidarum showed excellent fermentation performance utilizing glucose and xylose under anaerobic conditions. But this yeast is highly sensitive to the inhibitors such as furfural present in the pretreated lignocellulosic biomass. In order to improve...... from fusion of the protoplasts of S. passalidarum M7 and a robust yeast, Saccharomyces cerevisiae ATCC 96581, were able to grow in 75% WSLQ and produce around 0.4 g ethanol/g consumed xylose. Among the selected hybrid strains, the hybrid FS22 showed the best fermentation capacity in 75% WSLQ...... the inhibitor tolerance of this yeast, a combination of UV mutagenesis and protoplast fusion was used to construct strains with improved performance. Firstly, UVinduced mutants were screened and selected for improved tolerance towards furfural. The most promised mutant, S. passalidarum M7, produced 50% more...

  13. Voltammetric detection of phytochelatin transported across unmodified and protoplast modified model phospholipid membranes

    Czech Academy of Sciences Publication Activity Database

    Navrátil, Tomáš; Nováková, Kateřina; Josypčuk, Bohdan; Sokolová, Romana; Šestáková, Ivana

    2016-01-01

    Roč. 147, č. 1 (2016), s. 165-171 ISSN 0026-9247 R&D Projects: GA ČR(CZ) GAP208/12/1645 Grant - others:Rada Programu interní podpory projektů mezinárodní spolupráce AV ČR(CZ) M200401201 Program:M Institutional support: RVO:61388955 Keywords : Barley protoplasts * electrochemical impendance spectroscopy * mercury electrode Subject RIV: CG - Electrochemistry Impact factor: 1.282, year: 2016

  14. Influence of protoplast fusion between two Trichoderma spp. on extracellular enzymes production and antagonistic activity.

    Science.gov (United States)

    Hassan, Mohamed M

    2014-11-02

    Biological control plays a crucial role in grapevine pathogens disease management. The cell-wall degrading enzymes chitinase, cellulase and β-glucanase have been suggested to be essential for the mycoparasitism activity of Trichoderma species against grapevine fungal pathogens. In order to develop a useful strain as a single source of these vital enzymes, it was intended to incorporate the characteristics of two parental fungicides tolerant mutants of Trichoderma belonging to the high chitinase producing species T. harzianum and the high cellulase producing species T. viride , by fusing their protoplasts. The phylogeny of the parental strains was carried out using a sequence of the 5.8S-ITS region. The BLAST of the obtained sequence identified these isolates as T. harzianum and T. viride . Protoplasts were isolated using lysing enzymes and were fused using polyethylene glycol. The fused protoplasts have been regenerated on protoplast regeneration minimal medium supplemented with two selective fungicides. Among the 40 fast growing fusants, 17 fusants were selected based on their enhanced growth on selective media for further studies. The fusant strains were growing 60%-70% faster than the parents up to third generation. All the 17 selected fusants exhibited morphological variations. Some fusant strains displayed threefold increased chitinase enzyme activity and twofold increase in β-glucanase enzyme activity compared to the parent strains. Most fusants showed powerful antagonistic activity against Macrophomin aphaseolina , Pythium ultimum and Sclerotium rolfsii pathogens. Fusant number 15 showed the highest inhibition percentage (92.8%) against M. phaseolina and P. ultimum, while fusant number 9 showed the highest inhibition percentage (98.2%) against the growth of S. rolfsii. A hyphal intertwining and degradation phenomenon was observed by scanning electron microscope. The Trichoderma antagonistic effect against pathogenic fungal mycelia was due to the

  15. Transformation of haploid, microspore-derived cell suspension protoplasts of rice (Oryza sativa L.).

    Science.gov (United States)

    Chaïr, H; Legavre, T; Guiderdoni, E

    1996-06-01

    We compared the transient activity of three cereal gene-derived promoter-gus fusions and the efficiency of selection mediated by three different selectable genes in a polyethylene glycol transformation system with haploid cell suspension protoplasts of rice. The maize ubiquitin promoter was found to be the most active in transformed protoplasts, and selection on ammonium glufosinate mediated by the bar gene was the most efficient for producing resistant calluses. Cotransformation of protoplasts with two separate plasmids carrying the gus and the bar genes, at either a 2∶1 or 1∶1 ratio, led to 0.8 × 10(-5) and 1.6 × 10(-5) resistant callus recovery frequencies and 59.7 and 37.9 cotransformation efficiencies respectively. No escapes were detected in dot blot analyses of 100 resistant calluses with a probe consisting of the bar coding region. Cotransformation efficiency, based on resistance to basta and β-glucuronidase staining of the leaf tissue of 115 regenerated plants, was 47%. Resistance tests and Southern analysis of seed progenies of three diploid transgenic plants demonstrated homozygous integration of multiple copies of the transgene at one locus at least in the first plant, heterozygous integration at one locus in the second plant and heterozygous integration at two loci in the third plant.

  16. Effects of ultraviolet radiation on viability of isolated Beta vulgaris and Hordeum vulgare protoplasts

    International Nuclear Information System (INIS)

    Bornman, J.F.; Bjoern, L.O.; Bornman, C.H.

    1982-01-01

    Estimates of viability as measured by vital straining with fluorescein diacetate were carried out on freshly isolated and partially aged (16-hour-old) Beta vulgaris and Hordeum vulgare mesophyll protoplasts following irradiation with UV-B. Damage to the photosynthetic system by UV-B was determined by delayed light emission (DLE). In the case of freshly isolated Protoplasts Beta was approximately 30% more susceptible than Hordeum following 3h irradiation, with viability decreasing from 90% to 40%. After storage of protoplasts on ice for 16 h UV-B radiation markedly depressed viability in both species, but in the case of Hordeum there was a substantial initial loss of nearly 70% in viability over the first hour of irradiation. The first 10 min of UV-B radiation decreased the intensity of DLE by 40% without appreciably affecting the decay rate. Longer treatment times did not give a proportional effect so that even after 60 min of UV-B the inhibition did not exceed 60%. This suggested that although the enzyme system responsible for FDA hydrolysis may be partially inactivated (viability was 75-80% as compared with 90% in the control), the UV-B did not penetrate the innermost parts of the chloroplasts, but left some thylakoids undamaged. (orig.)

  17. Construction of Potent Recombinant Strain Through Intergeneric Protoplast Fusion in Endophytic Fungi for Anticancerous Enzymes Production Using Rice Straw.

    Science.gov (United States)

    El-Gendy, Mervat Morsy Abbas Ahmed; Al-Zahrani, Salha Hassan Mastour; El-Bondkly, Ahmed Mohamed Ahmed

    2017-09-01

    Among all fungal endophytes isolates derived from different ethno-medical plants, the hyper-yield L-asparaginase and L-glutaminase wild strains Trichoderma sp. Gen 9 and Cladosporium sp. Gen 20 using rice straw under solid-state fermentation (SSF) were selected. The selected strains were used as parents for the intergeneric protoplast fusion program to construct recombinant strain for prompt improvement production of these enzymes in one recombinant strain. Among 21 fusants obtained, the recombinant strain AYA 20-1, with 2.11-fold and 2.58-fold increase in L-asparaginase and L-glutaminase activities more than the parental isolates Trichoderma sp. Gen 9 and Cladosporium sp. Gen 20, respectively, was achieved using rice straw under SSF. Both therapeutic enzymes L-asparaginase and L-glutaminase were purified and characterized from the culture supernatant of the recombinant AYA 20-1 strain with molecular weights of 50.6 and 83.2 kDa, respectively. Both enzymes were not metalloenzymes. Whereas thiol group blocking reagents such as p-chloromercurybenzoate and iodoacetamide totally inhibited L-asparaginase activity, which refer to sulfhydryl groups and cysteine residues involved in its catalytic activity, they have no effect toward L-glutaminase activity. Interestingly, potent anticancer, antioxidant, and antimicrobial activities were detected for both enzymes.

  18. Application of Tissue Culture and Transformation Techniques in Model Species Brachypodium distachyon.

    Science.gov (United States)

    Sogutmaz Ozdemir, Bahar; Budak, Hikmet

    2018-01-01

    Brachypodium distachyon has recently emerged as a model plant species for the grass family (Poaceae) that includes major cereal crops and forage grasses. One of the important traits of a model species is its capacity to be transformed and ease of growing both in tissue culture and in greenhouse conditions. Hence, plant transformation technology is crucial for improvements in agricultural studies, both for the study of new genes and in the production of new transgenic plant species. In this chapter, we review an efficient tissue culture and two different transformation systems for Brachypodium using most commonly preferred gene transfer techniques in plant species, microprojectile bombardment method (biolistics) and Agrobacterium-mediated transformation.In plant transformation studies, frequently used explant materials are immature embryos due to their higher transformation efficiencies and regeneration capacity. However, mature embryos are available throughout the year in contrast to immature embryos. We explain a tissue culture protocol for Brachypodium using mature embryos with the selected inbred lines from our collection. Embryogenic calluses obtained from mature embryos are used to transform Brachypodium with both plant transformation techniques that are revised according to previously studied protocols applied in the grasses, such as applying vacuum infiltration, different wounding effects, modification in inoculation and cocultivation steps or optimization of bombardment parameters.

  19. Application of molecular techniques for the assessment of microorganism diversity on cultural heritage objects.

    Science.gov (United States)

    Otlewska, Anna; Adamiak, Justyna; Gutarowska, Beata

    2014-01-01

    As a result of their unpredictable ability to adapt to varying environmental conditions, microorganisms inhabit different types of biological niches on Earth. Owing to the key role of microorganisms in many biogeochemical processes, trends in modern microbiology emphasize the need to know and understand the structure and function of complex microbial communities. This is particularly important if the strategy relates to microbial communities that cause biodeterioration of materials that constitute our cultural heritage. Until recently, the detection and identification of microorganisms inhabiting objects of cultural value was based only on cultivation-dependent methods. In spite of many advantages, these methods provide limited information because they identify only viable organisms capable of growth under standard laboratory conditions. However, in order to carry out proper conservation and renovation, it is necessary to know the complete composition of microbial communities and their activity. This paper presents and characterizes modern techniques such as genetic fingerprinting and clone library construction for the assessment of microbial diversity based on molecular biology. Molecular methods represent a favourable alternative to culture-dependent methods and make it possible to assess the biodiversity of microorganisms inhabiting technical materials and cultural heritage objects.

  20. Response of haploid and diploid protoplasts from Datura innoxia Mill. and Petunia hybrida L. to treatment with X-rays and a chemical mutagen

    International Nuclear Information System (INIS)

    Krumbiegel, G.

    1979-01-01

    Haploid and diploid protoplasts of the two Solanaceous species Datura innoxia Mill. and Petunia Hybridia L., were exposed to two different mutagens, increased doses of X-rays and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). With both species the survival rates of haploid protoplasts decreased exponentially with increased doses of X-rays and increased concentrations of MNNG. Diploid protoplasts showed a higher resistance than haploids only at higher mutagen doses or concentrations. After the MNNG-treatment of haploid protoplasts from Datura innoxia, four mutants with altered pigment patterns were isolated. (author)

  1. Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation.

    Science.gov (United States)

    Li, Jian-Feng; Bush, Jenifer; Xiong, Yan; Li, Lei; McCormack, Matthew

    2011-01-01

    Protein-protein interactions (PPIs) constitute the regulatory network that coordinates diverse cellular functions. There are growing needs in plant research for creating protein interaction maps behind complex cellular processes and at a systems biology level. However, only a few approaches have been successfully used for large-scale surveys of PPIs in plants, each having advantages and disadvantages. Here we present split firefly luciferase complementation (SFLC) as a highly sensitive and noninvasive technique for in planta PPI investigation. In this assay, the separate halves of a firefly luciferase can come into close proximity and transiently restore its catalytic activity only when their fusion partners, namely the two proteins of interest, interact with each other. This assay was conferred with quantitativeness and high throughput potential when the Arabidopsis mesophyll protoplast system and a microplate luminometer were employed for protein expression and luciferase measurement, respectively. Using the SFLC assay, we could monitor the dynamics of rapamycin-induced and ascomycin-disrupted interaction between Arabidopsis FRB and human FKBP proteins in a near real-time manner. As a proof of concept for large-scale PPI survey, we further applied the SFLC assay to testing 132 binary PPIs among 8 auxin response factors (ARFs) and 12 Aux/IAA proteins from Arabidopsis. Our results demonstrated that the SFLC assay is ideal for in vivo quantitative PPI analysis in plant cells and is particularly powerful for large-scale binary PPI screens.

  2. Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation.

    Directory of Open Access Journals (Sweden)

    Jian-Feng Li

    Full Text Available Protein-protein interactions (PPIs constitute the regulatory network that coordinates diverse cellular functions. There are growing needs in plant research for creating protein interaction maps behind complex cellular processes and at a systems biology level. However, only a few approaches have been successfully used for large-scale surveys of PPIs in plants, each having advantages and disadvantages. Here we present split firefly luciferase complementation (SFLC as a highly sensitive and noninvasive technique for in planta PPI investigation. In this assay, the separate halves of a firefly luciferase can come into close proximity and transiently restore its catalytic activity only when their fusion partners, namely the two proteins of interest, interact with each other. This assay was conferred with quantitativeness and high throughput potential when the Arabidopsis mesophyll protoplast system and a microplate luminometer were employed for protein expression and luciferase measurement, respectively. Using the SFLC assay, we could monitor the dynamics of rapamycin-induced and ascomycin-disrupted interaction between Arabidopsis FRB and human FKBP proteins in a near real-time manner. As a proof of concept for large-scale PPI survey, we further applied the SFLC assay to testing 132 binary PPIs among 8 auxin response factors (ARFs and 12 Aux/IAA proteins from Arabidopsis. Our results demonstrated that the SFLC assay is ideal for in vivo quantitative PPI analysis in plant cells and is particularly powerful for large-scale binary PPI screens.

  3. Analysis of the Growth Process of Neural Cells in Culture Environment Using Image Processing Techniques

    Science.gov (United States)

    Mirsafianf, Atefeh S.; Isfahani, Shirin N.; Kasaei, Shohreh; Mobasheri, Hamid

    Here we present an approach for processing neural cells images to analyze their growth process in culture environment. We have applied several image processing techniques for: 1- Environmental noise reduction, 2- Neural cells segmentation, 3- Neural cells classification based on their dendrites' growth conditions, and 4- neurons' features Extraction and measurement (e.g., like cell body area, number of dendrites, axon's length, and so on). Due to the large amount of noise in the images, we have used feed forward artificial neural networks to detect edges more precisely.

  4. Bridging Mediterranean cultures in the IYS: A documentary exhibition on irrigation techniques in water scarcity conditions

    Science.gov (United States)

    Barontini, Stefano; Louki, Amina; Ben Slima, Zied; Ezzahra Ghaouch, Fatima; Labaran, Raisa; Raffelli, Giulia; Peli, Marco; Vitale, Nicola

    2015-04-01

    Brescia, an industrial city in Northern Italy, is now experiencing a crucial change in its traditional structure. In recent years in fact it has been elected as living and working seat by many foreigners and it is now one of the cities with the greatest percentage of migrants in the Country. This is an important challenge for the city and an opportunity to merge, compare and integrate different cultures to build its future. In this context some students of different Courses (engineering and medicine), belonging both to the Arabian and local community, met together and with researchers in the study team 'Al-B¯i r¯u n¯i , for culture, science and society'. The team aims at organising cultural events in which, starting from the figure of the Persian scientist Ab¯u Raih. ¯a n Al-B¯i r¯u n¯i (about 973, 1051), the contribution of the Arabian and Islamic culture to the development of the European one in the middle ages is investigated. Moving from the initial idea of the study team Al-B¯i r¯u n¯i and from the suggestions of the World Soil Day 2014 and of the International Year of Soils 2015, we built a documentary exhibition entitled 'Irrigation techniques in water scarcity conditions'. The exhibition, which stresses the importance of the irrigation techniques for the soil conservation, is focused on the idea of disseminating two main concepts, i.e. (1) the technological continuity of some water supply systems in countries, around the Mediterranean Sea, affected by similar conditions of water availability, and (2) the possibility of building environments where, due to severe or extreme climatic conditions, the sustainability is reached when the man lives in equilibrium with the nature. The exhibition, which is written in Italian and will move around in the city during all 2015, consists of about twenty posters organized into three main chapters, corresponding to three main classes of water supply systems which are common in most of the countries surrounding

  5. A Cultural Psychological Approach to Analyze Intercultural Learning: Potential and Limits of the Structure Formation Technique

    Directory of Open Access Journals (Sweden)

    Doris Weidemann

    2009-01-01

    Full Text Available Despite the huge interest in sojourner adjustment, there is still a lack of qualitative as well as of longitudinal research that would offer more detailed insights into intercultural learning processes during overseas stays. The present study aims to partly fill that gap by documenting changes in knowledge structures and general living experiences of fifteen German sojourners in Taiwan in a longitudinal, cultural-psychological study. As part of a multimethod design a structure formation technique was used to document subjective theories on giving/losing face and their changes over time. In a second step results from this study are compared to knowledge-structures of seven long-term German residents in Taiwan, and implications for the conceptualization of intercultural learning will be proposed. Finally, results from both studies serve to discuss the potential and limits of structure formation techniques in the field of intercultural communication research. URN: urn:nbn:de:0114-fqs0901435

  6. The use of tissue culture techniques with irradiation to improve potato resistance to late blight

    International Nuclear Information System (INIS)

    Al-Safadi, B.; Arabi, M.I.E.

    2004-01-01

    A mutation breeding program was conducted to improve potato (Solanum tuberosum) resistance to late blight disease caused by Phytophthora infestans. In vitro cultured explants from potato cvs. Draga, Diamant, Spunta were irradiated with gamma ray doses 25, 30, and 35 Gy. Growing shoots were cut and re-cultured every 2 weeks until the 4 t h generation (MV 4 ) to make sure no chimeral tissues still existed in the mutant material. Plantlets were subsequently propagated to obtain enough explants for in vitro selection pressure. Around 3000 plantlets from the three cultivars were subjected to selection pressure using co-culture technique. MV 4 explants were incubated in jars, containing MS medium, with mycelia of P. infestans. Surviving plantlets were propagated and re-incubated with the pathogen for three consecutive generations. Resistant plantlets were acclimatized and transferred to pots and grown under glasshouse conditions. Plants were later inoculated, at the adult stage, with sporangial suspension. Cultivar Draga produced the highest number of resistant plants. Ten plants of Draga appeared to be resistant to late blight whereas only one plant from each of the other 2 cultivars was resistant. Mutant plants varied in number of produced minitubers from 13 to 70, Also, weight of these minitubers varied from less than 1 to 35 grams. Selected mutant lines will undergo further testing under field conditions for P. infestans resistance and other agronomic characteristics. (author)

  7. Identification of Viable Helicobacter pylori in Drinking Water Supplies by Cultural and Molecular Techniques.

    Science.gov (United States)

    Santiago, Paula; Moreno, Yolanda; Ferrús, M Antonía

    2015-08-01

    Helicobacter pylori is one of the most common causes of chronic bacterial infection in humans, directly related to peptic ulcer and gastric cancer. It has been suggested that H. pylori can be acquired through different transmission routes, including water. In this study, culture and qPCR were used to detect and identify the presence of H. pylori in drinking water. Furthermore, the combined techniques PMA-qPCR and DVC-FISH were applied for detection of viable cells of H. pylori. Among 24 drinking water samples, 16 samples were positive for the presence of H. pylori, but viable cells were only detected in six samples. Characteristic colonies, covered by a mass of bacterial unspecific growth, were observed on selective agar plates from an only sample, after enrichment. The mixed culture was submitted to DVC-FISH and qPCR analysis, followed by sequencing of the amplicons. Molecular techniques confirmed the growth of H. pylori on the agar plate. Our results demonstrate for the first time that H. pylori can survive and be potentially infective in drinking water, showing that water distribution systems could be a potential route for H. pylori transmission. © 2015 John Wiley & Sons Ltd.

  8. Impact of accelerator-based analytical techniques on the knowledge and conservation of cultural heritage

    International Nuclear Information System (INIS)

    Dran, Jean Claude

    2001-01-01

    A large set of modern analytical techniques is currently applied to get a better insight on art and archaeological objects as well as to contribute to their conservation and restoration. Because of the precious and sometimes unique character of the works, non-destructive techniques and even those requiring no (or only minute) sampling, are preferred. From this standpoint, ion beam analysis (IBA) constitutes one of the best choices, since it combines quite good analytical performance and non-destructiveness. For over 10 years, an IBA facility has been installed in the Research Laboratory of the Museums of France. Until now it is the only facility of this kind entirely devoted to the study of cultural heritage. A special set-up, namely an external beam line, has been developed which permits the in-air analysis of large or fragile works of art without sampling. This facility is used for both short investigations at the request of museum curators and extensive research works in art history and archaeology. Numerous examples will be given to highlight the impact of this tool on cultural heritage. (author)

  9. Spatial Techniques to Visualize Acoustic Comfort along Cultural and Heritage Routes for a World Heritage City

    Directory of Open Access Journals (Sweden)

    Ni Sheng

    2015-07-01

    Full Text Available This paper proposes to visualize acoustic comfort along tourist routes. Route-based tourism is crucial to the sustainability of tourism development in historic areas. Applying the concept of route-based tourism to guide tourists rambling along cultural and heritage routes can relieve overcrowded condition at hot scenic spots and increase the overall carrying capacity of the city. However, acoustic comfort along tourist routes is rarely addressed in academic studies and decision-making. Taking Macao as an example, this paper has studied pedestrian exposure to traffic noise along the cultural and heritage routes. The study is based on a GIS-based traffic noise model system with a high spatial resolution down to individual buildings along both sides of the street. Results show that tourists suffer from excessive traffic noise at certain sites, which may have negative impact on the promotion of route-based tourism in the long run. In addition, it is found that urban growth affects urban form and street layout, which in turn affect traffic flow and acoustic comfort in urban area. The present study demonstrates spatial techniques to visualize acoustic comfort along tourist routes, and the techniques are foreseen to be used more frequently to support effective tourism planning in the future.

  10. Mutation breeding for disease resistance using in-vitro culture techniques

    International Nuclear Information System (INIS)

    1985-07-01

    Breeding for disease resistance is a major aspect of plant breeding, which may take at least 20% of a plant breeder's time, effort and budget. Nevertheless, numerous resistance problems remain unsolved and present major constraints to the production of food, feed, fiber and industrial commodities. The application of novel biotechnology and genetic engineering will extend the possibilities of conventional plant breeding. Therefore a meeting of experts in plant protection, plant breeding and in-vitro culture technology was convened by the Joint FAO/IAEA Division in Vienna. The experts were asked to discuss and give advice on prospects of biotechnology, especially plant in-vitro cultures, to contribute towards improved chances of success in mutation breeding for disease resistance. The plant breeder, in searching for resistance to a particular pathogen, like for any other desirable character, needs genetic variation to begin with. In addition he needs an appropriate screening method to detect the desired character. Science has developed so fast that it is now time to apply the existing knowledge of biotechnology to practical problems in agriculture, also in developing countries. In the near future this may be true also for novel techniques of genetic engineering. The usefulness and feasibility of the application of in-vitro techniques for these purposes varies with crops and pathogens, but also depends on the strength of plant breeding and plant pathology and the facilities available in a particular country. The members of the Advisory Group attempted to discuss the various aspects and to reach sound conclusions

  11. Somatic Embryogenesis: Still a Relevant Technique in Citrus Improvement.

    Science.gov (United States)

    Omar, Ahmad A; Dutt, Manjul; Gmitter, Frederick G; Grosser, Jude W

    2016-01-01

    The genus Citrus contains numerous fresh and processed fruit cultivars that are economically important worldwide. New cultivars are needed to battle industry threatening diseases and to create new marketing opportunities. Citrus improvement by conventional methods alone has many limitations that can be overcome by applications of emerging biotechnologies, generally requiring cell to plant regeneration. Many citrus genotypes are amenable to somatic embryogenesis, which became a key regeneration pathway in many experimental approaches to cultivar improvement. This chapter provides a brief history of plant somatic embryogenesis with focus on citrus, followed by a discussion of proven applications in biotechnology-facilitated citrus improvement techniques, such as somatic hybridization, somatic cybridization, genetic transformation, and the exploitation of somaclonal variation. Finally, two important new protocols that feature plant regeneration via somatic embryogenesis are provided: protoplast transformation and Agrobacterium-mediated transformation of embryogenic cell suspension cultures.

  12. Withania somnifera: Advances and Implementation of Molecular and Tissue Culture Techniques to Enhance Its Application

    Directory of Open Access Journals (Sweden)

    Vibha Pandey

    2017-08-01

    Full Text Available Withania somnifera, commonly known as Ashwagandha an important medicinal plant largely used in Ayurvedic and indigenous medicine for over 3,000 years. Being a medicinal plant, dried powder, crude extract as well as purified metabolies of the plant has shown promising therapeutic properties. Withanolides are the principal metabolites, responsible for the medicinal properties of the plant. Availability and amount of particular withanolides differ with tissue type and chemotype and its importance leads to identification characterization of several genes/ enzymes related to withanolide biosynthetic pathway. The modulation in withanolides can be achieved by controlling the environmental conditions like, different tissue culture techniques, altered media compositions, use of elicitors, etc. Among all the in vitro techniques, hairy root culture proved its importance at industrial scale, which also gets benefits due to more accumulation (amount and number of withanolides in roots tissues of W. somnifera. Use of media compostion and elicitors further enhances the amount of withanolides in hairy roots. Another important modern day technique used for accumulation of desired secondary metabolites is modulating the gene expression by altering environmental conditions (use of different media composition, elicitors, etc. or through genetic enginnering. Knowing the significance of the gene and the key enzymatic step of the pathway, modulation in withanolide contents can be achieved upto required amount in therapeutic industry. To accomplish maximum productivity through genetic enginnering different means of Withania transformation methods have been developed to obtain maximum transformation efficiency. These standardized transformation procedues have been used to overexpress/silence desired gene in W. somnifera to understand the outcome and succeed with enhanced metabolic production for the ultimate benefit of human race.

  13. Development of a cell culture surface conversion technique using alginate thin film for evaluating effect upon cellular differentiation

    International Nuclear Information System (INIS)

    Nakashima, Y.; Tsusu, K.; Minami, K.; Nakanishi, Y.

    2014-01-01

    Here, we sought to develop a cell culture surface conversion technique that would not damage living cells. An alginate thin film, formed on a glass plate by spin coating of sodium alginate solution and dipping into calcium chloride solution, was used to inhibit adhesion of cells. The film could be removed by ethylenediaminetetraacetate (EDTA) at any time during cell culture, permitting observation of cellular responses to conversion of the culture surface in real time. Additionally, we demonstrated the validity of the alginate thin film coating method and the performance of the film. The thickness of the alginate thin film was controlled by varying the rotation speed during spin coating. Moreover, the alginate thin film completely inhibited the adhesion of cultured cells to the culture surface, irrespective of the thickness of the film. When the alginate thin film was removed from the culture surface by EDTA, the cultured cells adhered to the culture surface, and their morphology changed. Finally, we achieved effective differentiation of C2C12 myoblasts into myotube cells by cell culture on the convertible culture surface, demonstrating the utility of our novel technique

  14. Transposition of a Ds element from a plasmid into the plant genome in Nicotiana plumbaginifolia protoplast-derived cells.

    Science.gov (United States)

    Houba-Hérin, N; Domin, M; Pédron, J

    1994-07-01

    Nicotiana plumbaginifolia haploid protoplasts were co-transformed with two plasmids, one with a NPT-II/Ds element and one with a gene encoding an amino-terminal truncated Ac transposase. It is shown that Ds can efficiently transpose from extrachromosomal DNA to N. plumbaginifolia chromosomes when the Ac transposase gene is present in trans. Ds has been shown to have transposed into the plant genome in a limited number of copies (1.9 copies per genome), for 21/32 transgenic lines tested. The flanking sequences present in the original plasmid are missing in these 21 plants. In only two of 21 plants was part of the transposase construct integrated. By segregation analysis of transgenic progeny, Ds was shown to be present in the heterozygous state in 10 lines even though haploid protoplasts had been originally transformed. This observation could indicate that integration occurred after or during DNA replication that leads to protoplast diploidization.

  15. Extending the fungal host range of a partitivirus and a mycoreovirus from Rosellinia necatrix by inoculation of protoplasts with virus particles.

    Science.gov (United States)

    Kanematsu, Satoko; Sasaki, Atsuko; Onoue, Mari; Oikawa, Yuri; Ito, Tsutae

    2010-09-01

    The potential host range of mycoviruses is poorly understood because of the lack of suitable inoculation methods. Recently, successful transfection has been reported for somatically incompatible fungal isolates with purified virus particles of two mycoviruses, the partitivirus RnPV1-W8 (RnPV1) and the mycoreovirus RnMyRV3/W370 (MyRV3), from the white root rot fungus Rosellinia necatrix (class Sordariomycetes, subclass Xylariomycetidae). These studies examined and revealed the effect of the mycoviruses on growth and pathogenicity of R. necatrix. Here, we extended the experimental host range of these two mycoviruses using a transfection approach. Protoplasts of other phytopathogenic Sordariomycetous fungi-Diaporthe sp., Cryphonectria parasitica, Valsa ceratosperma (Sordariomycetidae), and Glomerella cingulata (Hypocreomycetidae)-were inoculated with RnPV1 and MyRV3 viral particles. The presence of double-stranded RNA viral genomes in regenerated mycelia of Diaporthe sp., C. parasitica, and V. ceratosperma confirmed both types of viral infections in these three novel host species. An established RnPV1 infection was confirmed in G. cingulata but MyRV3 did not infect this host. Horizontal transmission of both viruses from newly infected strains to virus-free, wild-type strains through hyphal anastomosis was readily achieved by dual culture; however, vertical transmission through conidia was rarely observed. The virulence of Diaporthe sp., C. parasitica, and V. ceratosperma strains harboring MyRV3 was reduced compared with their virus-free counterpart. In summary, our protoplast inoculation method extended the experimental host range of RnPV1-W8 and MyRV3 within the class Sordariomycetes and revealed that MyRV3 confers hypovirulence to the new hosts, as it does to R. necatrix.

  16. The potential of 3D techniques for cultural heritage object documentation

    Science.gov (United States)

    Bitelli, Gabriele; Girelli, Valentina A.; Remondino, Fabio; Vittuari, Luca

    2007-01-01

    The generation of 3D models of objects has become an important research point in many fields of application like industrial inspection, robotics, navigation and body scanning. Recently the techniques for generating photo-textured 3D digital models have interested also the field of Cultural Heritage, due to their capability to combine high precision metrical information with a qualitative and photographic description of the objects. In fact this kind of product is a fundamental support for documentation, studying and restoration of works of art, until a production of replicas by fast prototyping techniques. Close-range photogrammetric techniques are nowadays more and more frequently used for the generation of precise 3D models. With the advent of automated procedures and fully digital products in the 1990s, it has become easier to use and cheaper, and nowadays a wide range of commercial software is available to calibrate, orient and reconstruct objects from images. This paper presents the complete process for the derivation of a photorealistic 3D model of an important basalt stela (about 70 x 60 x 25 cm) discovered in the archaeological site of Tilmen Höyük, in Turkey, dating back to 2nd mill. BC. We will report the modeling performed using passive and active sensors and the comparison of the achieved results.

  17. Applications of synchrotron-based micro-imaging techniques for the analysis of Cultural Heritage materials

    International Nuclear Information System (INIS)

    Cotte, Marine; Chilida, Javier; Walter, Philippe; Taniguchi, Yoko; Susini, Jean

    2009-01-01

    The analysis of cultural Heritage objects is often technically challenging. When analyzing micro-fragments, the amount of matter is usually very tiny, hence requiring sensitive techniques. These samples, in particular painting fragments, may present multi-layered structures, with layer thickness of ∼10 μm. It leads to favor micro-imaging techniques, with a good lateral resolution (about one micrometer), that manage the discriminative study of each layer. Besides, samples are usually very complex in term of chemistry, as they are made of mineral and organic matters, amorphous and crystallized phases, major and minor elements. Accordingly, a multi-modal approach is generally essential to solve the chemical complexity of such hybrid materials. Different examples will be given, to illustrate the various possibilities of synchrotron-based micro-imaging techniques, such as micro X-ray diffraction, micro X-ray fluorescence, micro X-ray absorption spectroscopy and micro FTIR spectroscopy. Focus will be made on paintings, but the whole range of museum objects (going from soft matter like paper or wood to hard matter like metal and glass) will be also considered.

  18. Application of culture dependent and cultural-independent techniques to investigate the dynamics of microorganisms during industrial cheese making of a Gouda type cheese

    OpenAIRE

    Perolari, Alessandra

    2012-01-01

    The aim of this work was to study the microbial dynamic in a Gouda type cheese applying a comparison between culture-dependent technique, plate count, and culture-independent technique as PCR-DGGE. The study was also focus on the analyse of the efficiency of the pasteurizer, and its cleaning steps. The study was conducted analysing 4 batches for each of the two days of production, Monday and Friday. Between batch 16 and 17 there was a quick cleaning, and between batch 32 and batch 1, of th...

  19. Chondrogenesis of human adipose derived stem cells for future microtia repair using co-culture technique.

    Science.gov (United States)

    Goh, Bee See; Che Omar, Siti Nurhadis; Ubaidah, Muhammad Azhan; Saim, Lokman; Sulaiman, Shamsul; Chua, Kien Hui

    2017-04-01

    In conclusion, these result showed HADSCs could differentiate into chondrocytes-like cells, dependent on signaling induced by TGF-β3 and chondrocytes. This is a promising result and showed that HADSCs is a potential source for future microtia repair. The technique of co-culture is a positive way forward to assist the microtia tissue. Reconstructive surgery for the repair of microtia still remains the greatest challenge among the surgeons. Its repair is associated with donor-site morbidity and the degree of infection is inevitable when using alloplastic prosthesis with uncertain long-term durability. Thus, human adipose derived stem cells (HADSCs) can be an alternative cell source for cartilage regeneration. This study aims to evaluate the chondrogenic potential of HADSCs cultured with transforming growth factor-beta (TGF-β) and interaction of auricular chondrocytes with HADSCs for new cartilage generation. Multi-lineages differentiation features of HADSCs were monitored by Alcian Blue, Alizarin Red, and Oil Red O staining for chondrogenic, adipogenic, and osteogenic differentiation capacity, respectively. Further, HADSCs alone were culture in medium added with TGF-β3; and human auricular chondrocytes were interacted indirectly in the culture with and without TGF-βs for up to 21 days, respectively. Cell morphology and chondrogenesis were monitored by inverted microscope. For cell viability, Alamar Blue assay was used to measure the cell viability and the changes in gene expression of auricular chondrocyte markers were determined by real-time polymerase chain reaction analysis. For the induction of chondrogenic differentiation, HADSCs showed a feature of aggregation and formed a dense matrix of proteoglycans. Staining results from Alizirin Red and Oil Red O indicated the HADSCs also successfully differentiated into adipogenic and osteogenic lineages after 21 days. According to a previous study, HADSCs were strongly positive for the mesenchymal markers CD90, CD73

  20. In vitro culture of higher plants as a tool in the propagation of horticultural crops.

    NARCIS (Netherlands)

    Pierik, R.L.M.

    1988-01-01

    In vitro culture of higher plants is the culture, under sterile conditions, of plants, seeds, embryos, organs, explants, tissues, cells and protoplasts on nutrient media. This type of culture has shown spectacular development since 1975, resulting in the production and regeneration of viable

  1. The characterization of artefacts of cultural heritage significance using physical techniques

    International Nuclear Information System (INIS)

    Creagh, D.C.

    2005-01-01

    All societies attempt to preserve their cultural heritage because it is this that gives them their identity. How artefacts are identified as being of significance to society, and how to preserve these for posterity, depend on the sophistication of those societies, their wealth, and the determination of members of the societies to preserve their past. If conservation or restoration measures are being undertaken complex analytical experiments must be undertaken beforehand to ensure that the work is being undertaken in an appropriate manner. These investigations may employ electromagnetic (IR, VIS, UV, X-ray, γ-ray) or particulate (electron, proton, neutron, and ion beams) radiation. The use of many of these techniques is described in this paper in experiments on Australian Aboriginal bark paintings, a suit of armour belonging to a famous Australian outlaw, and the degradation of colour motion picture film

  2. Properties of Single K+ and Cl− Channels in Asclepias tuberosa Protoplasts 1

    Science.gov (United States)

    Schauf, Charles L.; Wilson, Kathryn J.

    1987-01-01

    Potassium and chloride channels were characterized in Asclepias tuberosa suspension cell derived protoplasts by patch voltage-clamp. Whole-cell currents and single channels in excised patches had linear instantaneous current-voltage relations, reversing at the Nernst potentials for K+ and Cl−, respectively. Whole cell K+ currents activated exponentially during step depolarizations, while voltage-dependent Cl− channels were activated by hyperpolarizations. Single K+ channel conductance was 40 ± 5 pS with a mean open time of 4.5 milliseconds at 100 millivolts. Potassium channels were blocked by Cs+ and tetraethylammonium, but were insensitive to 4-aminopyridine. Chloride channels had a single-channel conductance of 100 ± 17 picosiemens, mean open time of 8.8 milliseconds, and were blocked by Zn2+ and ethacrynic acid. Whole-cell Cl− currents were inhibited by abscisic acid, and were unaffected by indole-3-acetic acid and 2,4-dichlorophenoxyacetic acid. Since internal and external composition can be controlled, patch-clamped protoplasts are ideal systems for studying the role of ion channels in plant physiology and development. Images Fig. 5 PMID:16665712

  3. Analysis of Microbe-Associated Molecular Pattern-Responsive Synthetic Promoters with the Parsley Protoplast System.

    Science.gov (United States)

    Kanofsky, Konstantin; Lehmeyer, Mona; Schulze, Jutta; Hehl, Reinhard

    2016-01-01

    Plants recognize pathogens by microbe-associated molecular patterns (MAMPs) and subsequently induce an immune response. The regulation of gene expression during the immune response depends largely on cis-sequences conserved in promoters of MAMP-responsive genes. These cis-sequences can be analyzed by constructing synthetic promoters linked to a reporter gene and by testing these constructs in transient expression systems. Here, the use of the parsley (Petroselinum crispum) protoplast system for analyzing MAMP-responsive synthetic promoters is described. The synthetic promoter consists of four copies of a potential MAMP-responsive cis-sequence cloned upstream of a minimal promoter and the uidA reporter gene. The reporter plasmid contains a second reporter gene, which is constitutively expressed and hence eliminates the requirement of a second plasmid used as a transformation control. The reporter plasmid is transformed into parsley protoplasts that are elicited by the MAMP Pep25. The MAMP responsiveness is validated by comparing the reporter gene activity from MAMP-treated and untreated cells and by normalizing reporter gene activity using the constitutively expressed reporter gene.

  4. Organization of cytoskeleton controls the changes in cytosolic calcium of cold-shocked Nicotiana plumbaginifolia protoplasts.

    Science.gov (United States)

    Mazars, C; Thion, L; Thuleau, P; Graziana, A; Knight, M R; Moreau, M; Ranjeva, R

    1997-11-01

    Using Nicotiana plumbaginifolia constitutively expressing the recombinant bioluminescent calcium indicator, aequorin, it has been previously demonstrated that plant cells react to cold-shock by an immediate rise in cytosolic calcium. Such an opportune system has been exploited to address the regulatory pathway involved in the calcium response. For this purpose, we have used protoplasts derived from N. plumbaginifolia leaves that behave as the whole plant but with a better reproducibility. By both immunodetecting cytoskeletal components on membrane ghosts and measuring the relative change in cytosolic calcium, we demonstrate that the organization of the cytoskeleton has profound influences on the calcium response. The disruption of the microtubule meshwork by various active drugs, such as colchicin, oryzalin and vinblastin, leads to an important increase in the cytosolic calcium (up to 400 nM) in cold-shocked protoplasts over control. beta-Lumicolchicin, an inactive analogue of colchicin, is ineffective either on cytoplasmic calcium increase or on microtubule organization. A microfilament disrupting drug, cytochalasin D, exerts a slight stimulatory effect, whereas the simultaneous disruption of microtubule and microfilament meshworks results in a dramatic increase in the calcium response to cold-shock. The results described in the present paper illustrate the role of the intracellular organization and, more specifically, the role of cytoskeleton in controlling the intensity of calcium response to an extracellular stimulus.

  5. A Protoplast Transient Expression System to Enable Molecular, Cellular, and Functional Studies in Phalaenopsis orchids

    Directory of Open Access Journals (Sweden)

    Hsiang-Yin Lin

    2018-06-01

    Full Text Available The enigmatic nature of the specialized developmental programs of orchids has fascinated plant biologists for centuries. The recent releases of orchid genomes indicate that orchids possess new gene families and family expansions and contractions to regulate a diverse suite of developmental processes. However, the extremely long orchid life cycle and lack of molecular toolkit have hampered the advancement of orchid biology research. To overcome the technical difficulties and establish a platform for rapid gene regulation studies, in this study, we developed an efficient protoplast isolation and transient expression system for Phalaenopsis aphrodite. This protocol was successfully applied to protein subcellular localization and protein–protein interaction studies. Moreover, it was confirmed to be useful in delineating the PaE2F/PaDP-dependent cell cycle pathway and studying auxin response. In summary, the established orchid protoplast transient expression system provides a means to functionally characterize orchid genes at the molecular level allowing assessment of transcriptome responses to transgene expression and widening the scope of molecular studies in orchids.

  6. IBA techniques: Examples of useful combinations for the characterisation of cultural heritage materials

    Science.gov (United States)

    Beck, L.; Pichon, L.; Moignard, B.; Guillou, T.; Walter, P.

    2011-12-01

    For many years, ion beam analysis techniques have successfully been used to the study of cultural heritage objects. The chemical composition of work art is usually determined by PIXE, but in many cases, RBS and/or PIGE can provide useful complementary information. RBS gives information about the depth distribution and concentration in light elements, such as carbon and oxygen. In the past years, the experimental facilities at the AGLAE (Accélérateur Grand Louvre d'Analyse Élémentaire) accelerator has been progressively developed in order to apply simultaneously PIXE, PIGE and RBS under optimal conditions using an external beam. This combination is now routinely used for point analyses or mappings. In this contribution, we present several examples of applications: manufacturing technology of lustre-decorated ceramics and silver plating, control of altered or restored surfaces, and quantification of organic phase in painting and bone. The final conclusion is that the association of PIXE with RBS is very attractive for the investigation of cultural heritage objects, in particular of materials containing both mineral and organic components or possessing a multilayered structure. The first results of the production of monochromatic X-rays for radiography purposes by PIXE are also presented.

  7. IBA techniques: Examples of useful combinations for the characterisation of cultural heritage materials

    International Nuclear Information System (INIS)

    Beck, L.; Pichon, L.; Moignard, B.; Guillou, T.; Walter, P.

    2011-01-01

    For many years, ion beam analysis techniques have successfully been used to the study of cultural heritage objects. The chemical composition of work art is usually determined by PIXE, but in many cases, RBS and/or PIGE can provide useful complementary information. RBS gives information about the depth distribution and concentration in light elements, such as carbon and oxygen. In the past years, the experimental facilities at the AGLAE (Accélérateur Grand Louvre d’Analyse Élémentaire) accelerator has been progressively developed in order to apply simultaneously PIXE, PIGE and RBS under optimal conditions using an external beam. This combination is now routinely used for point analyses or mappings. In this contribution, we present several examples of applications: manufacturing technology of lustre-decorated ceramics and silver plating, control of altered or restored surfaces, and quantification of organic phase in painting and bone. The final conclusion is that the association of PIXE with RBS is very attractive for the investigation of cultural heritage objects, in particular of materials containing both mineral and organic components or possessing a multilayered structure. The first results of the production of monochromatic X-rays for radiography purposes by PIXE are also presented.

  8. Induced mutation and in vitro culture techniques for the genetic improvement of ornamentals

    International Nuclear Information System (INIS)

    Lapade, Avelina G.; Veluz, Ana Maria S.; Marbella, Lucia J.; Rama, Manny G.

    2001-01-01

    Mutation breeding using cobalt-60 ( 60 Co) gamma radiation coupled with tissue culture techniques is undertaken for genetic improvement of foliage ornamentals (Dracaena sp. and Murraya exotica L.) and cutflowers (Chrysanthemum morifolium and orchids; Vanda sanderiana, Dendrobium Pattaya Beauty and Phalenopsis schilleriana). Gamma radiation (10-30 Gy) induced chlorophyll mutations and several morphological changes in D. sanderiana. For D. godseffiana, irradiated cuttings resulted in reduction of leaf size and chlorophyll mutations. Reduction in height was observed in the M 2 generation of Murraya exotica L. irradiated at doses ranging from 10 to 30 Gy. The dwarf Murraya mutant was multiplied through the use of seeds and presently 116 plants are commercially available and are ''test marketed'' to the public. Tissue culture technique was used to induce mutation and as a means of micropropagation in two ornamental crops (orchids and chrysanthemum). Effects of different doses of gamma radiation on callus induction from nodal sections of chrysanthemum grown in Murashige and Skoog's (MS) with naphthalene acetic acid (NAA) and benzyl adenine (BA) were studied. Micropropagation of irradiated and unirradiated chrysanthemum using MS basal medium is presently being studied. Whorling and changes in leaf color were observed at 10 Gy and doubling of leaf growth at the node at 20 Gy for vegetatively generated V 3 plant. In orchids, irradiation of immature embryo with gamma rays ranging from 5 to 10 Gy increased the percentage of germination in Dendrobium Pattaya Beauty and P. schilleriana. Protocorms of Vanda sanderiana irradiated at 10 Gy and grown in Knudson C medium developed into plantlets that are bigger and more vigorous than those irradiated at 20 GY and from the control plant. A decrease in seedling height was observed with increasing dose of gamma radiation. (Author)

  9. X-ray tomographic techniques for the study of cultural heritages

    International Nuclear Information System (INIS)

    Schillaci, T.

    2011-01-01

    In recent years, X-ray Computed Tomography (CT) has become an important tool for investigating all kinds of materials. Due to its non-destructive nature, it is especially suitable to investigate samples that may not be altered or damaged during the course of the investigation. CT has been recently introduced in the field of Cultural Heritage diagnostics, where it can be used for the investigation of different works of art, as it preserves the integrity of the object and gives morphological and physical information on its inner structure. This paper describes a methodological approach on the use of the X-ray CT technique to study items belonging to cultural heritage with the aim to obtain information related to their preservation state and therefore, to plan an adequate conservation and restoration procedure. Significant examples of applications are the study of porosity and pore size distribution and their connectivity for different porous materials and the study of kinetics of capillary fluid absorption in sedimentary rocks. Other applications are relevant to the possibility to investigate in a non-destructive way the presence of defects or fractures inside an object and, not last in order of importance, the possibility to study different typologies of woods or waterlogged woods, the presence of an eventual biodegradation state and the possibility to perform a dendrochronology. In this paper, the results of some case studies, obtained through the integrated use of CT systems with different resolutions, are reported. Other expected future developments will be addressed to the integration of CT data with results of compatible non-destructive techniques.

  10. Drifter technique: a new method to obtain metaphases in Hep-2 cell line cultures

    Directory of Open Access Journals (Sweden)

    Eleonidas Moura Lima

    2005-07-01

    Full Text Available The Hep-2 cell line is derived from laryngeal carcinoma cells and is often utilized as a model in carcinogenesis and mutagenesis tests. To evaluate the proliferative potential of this line, we developed a cytogenetic methodology (drifter technique to obtain metaphases from cells that loose cellular adhesion when they underwent mitosis in culture. By this procedure, 2000 cells were counted, resulting in a mitotic index (MI of 22.2%. Although this MI was not statistically different from the one obtained using either a classical cytogenetic method or a cell synchronization technique, the drifter technique has the advantage of not requiring the use of some reagents for the obtention of metaphases and also of diminishing the consumption of maintenance reagents for this cell line.A linhagem celular Hep-2 é formada por células de carcinoma da laringe e é muito utilizada em modelos de carcinogênese e mutagenêse. Para avaliar o potencial proliferativo desta linhagem, desenvolvemos uma metodologia citogenética (técnica do sobrenadante para obtenção de metáfases a partir de células que, ao entrarem em mitose, perdem adesão celular, ficando em suspensão no meio de cultura. Através deste procedimento, foram contadas 2000 células, correspondendo a um índice mitótico (IM de 22.2% . Apesar de o IM obtido por esta técnica não ter sido estatisticamente diferente do IM obtido por outras metodologias citogenéticas clássicas, a técnica do sobrenadante é vantajosa porque elimina o uso de alguns reagentes utilizados na obtenção de metáfases e também diminui o consumo de reagentes de manutenção desta linhagem.

  11. Challenges, Strategies and Techniques for International Training in Technology for Cultural Heritage Conservation

    Science.gov (United States)

    Eppich, R.; Almagro Vidal, A.

    2013-07-01

    , Jordan, Argentina, United Arab Emirates, United States of America and around 20 other countries. These strategies deal with establishing methodologies and guiding principles for the selection of technologies, highlighting successful illustrated examples, levelling uneven educational bases and gaining access to expertise. The authors have developed these strategies and techniques to appeal, engage and succeed with such diverse groups - to encourage the participants to cooperate on a common goal and overcome specific challenges while embracing the technology and thinking critically about its appropriate application for the conservation of cultural heritage in their home countries. Other strategies include setting norms that respect the various cultures and differing levels of technology education, offering voluntary sessions for more advanced and ambitious participants, finding and then adopting natural leaders as co-instructors and offering a mix of sessions including standard lectures combined with field and laboratory exercises and distance learning. This methodology and strategies have proven to be successful as the participants have provided positive evaluations months and/or years after the courses, implemented their own courses using the materials and methods and have established a strong, sustainable network related to this topic.

  12. Probing plasma membrane microdomains in cowpea protoplasts using lipidated GFP-fusion proteins and multimode FRET microscopy

    NARCIS (Netherlands)

    Vermeer, J.E.M.; van Munster, E.B.; Vischer, N.O.; Gadella, T.

    2004-01-01

    Multimode fluorescence resonance energy transfer (FRET) microscopy was applied to study the plasma membrane organization using different lipidated green fluorescent protein (GFP)-fusion proteins co-expressed in cowpea protoplasts. Cyan fluorescent protein (CFP) was fused to the hyper variable region

  13. Early activation of lipoxygenase in lentil (Lens culinaris) root protoplasts by oxidative stress induces programmed cell death

    NARCIS (Netherlands)

    Vliegenthart, J.F.G.; Maccarrone, M.; Zadelhoff, G. van; Veldink, G.A.; Finazzi Agrò, A.

    2000-01-01

    Oxidative stress caused by hydrogen peroxide (H2O2) triggers the hypersensitive response of plants to pathogens. Here, short pulses of H2O2 are shown to cause death of lentil (Lens culinaris) root protoplasts. Dead cells showed DNA fragmentation and ladder formation, typical hallmarks of apoptosis

  14. Studies on protein synthesis by protoplasts of Saccharomyces carlsbergensis I. The effect of ribonuclease on protein synthesis

    NARCIS (Netherlands)

    Kloet, S.R. de; Wermeskerken, R.K.A. van; Koningsberger, V.V.

    1961-01-01

    Ribonuclease was found to inhibit the protein synthesis in the naked yeast protoplast for nearly 100%. Even small concentrations (5 μg/ml) were found inhibitory. The cause of this inhibition can be attributed at least in part to a 90% inhibition of the respiration. Amino acid uptake was found to

  15. Comparison of group B streptococci colonization in vaginal and rectal specimens by culture method and polymerase chain reaction technique

    Directory of Open Access Journals (Sweden)

    Shahrokh Bidgani

    2016-03-01

    Conclusion: The frequency of GBS culture from rectal samples was higher than vaginal samples. However, the detection percentage of GBS using PCR from vaginal samples was higher than rectal samples. By contrast, the culture is a time-consuming method requiring at least 48 hours for GBS fully identification but PCR is a sensitive and rapid technique in detection of GBS, with the result was acquired during 3 hours.

  16. Investigating the diversity of Pseudomonas spp. in soil using culture dependent and independent techniques

    DEFF Research Database (Denmark)

    Li, Lili; Abu Al-Soud, Waleed; Bergmark, Lasse

    2013-01-01

    Less than 1% of bacterial populations present in environmental samples are culturable, meaning that cultivation will lead to an underestimation of total cell counts and total diversity. However, it is less clear whether this is also true for specific well-defined groups of bacteria for which sele...... selective culture media is available. In this study, we use culture dependent and independent techniques to describe whether isolation of Pseudomonas spp. on selective nutrient-poor NAA 1: 100 agar-medium can reflect the full diversity, found by ...

  17. Impact of changing from staining to culture techniques on detection rates of Campylobacter spp. in routine stool samples in Chile.

    Science.gov (United States)

    Porte, Lorena; Varela, Carmen; Haecker, Thomas; Morales, Sara; Weitzel, Thomas

    2016-05-13

    Campylobacter is a leading cause of bacterial gastroenteritis, but sensitive diagnostic methods such as culture are expensive and often not available in resource limited settings. Therefore, direct staining techniques have been developed as a practical and economical alternative. We analyzed the impact of replacing Campylobacter staining with culture for routine stool examinations in a private hospital in Chile. From January to April 2014, a total of 750 consecutive stool samples were examined in parallel by Hucker stain and Campylobacter culture. Isolation rates of Campylobacter were determined and the performance of staining was evaluated against culture as the gold standard. Besides, isolation rates of Campylobacter and other enteric pathogens were compared to those of past years. Campylobacter was isolated by culture in 46 of 750 (6.1 %) stool samples. Direct staining only identified three samples as Campylobacter positive and reached sensitivity and specificity values of 6.5 and 100 %, respectively. In comparison to staining-based detection rates of previous years, we observed a significant increase of Campylobacter cases in our patients. Direct staining technique for Campylobacter had a very low sensitivity compared to culture. Staining methods might lead to a high rate of false negative results and an underestimation of the importance of campylobacteriosis. With the inclusion of Campylobacter culture, this pathogen became a leading cause of intestinal infection in our patient population.

  18. Using Web-Based Knowledge Extraction Techniques to Support Cultural Modeling

    Science.gov (United States)

    Smart, Paul R.; Sieck, Winston R.; Shadbolt, Nigel R.

    The World Wide Web is a potentially valuable source of information about the cognitive characteristics of cultural groups. However, attempts to use the Web in the context of cultural modeling activities are hampered by the large-scale nature of the Web and the current dominance of natural language formats. In this paper, we outline an approach to support the exploitation of the Web for cultural modeling activities. The approach begins with the development of qualitative cultural models (which describe the beliefs, concepts and values of cultural groups), and these models are subsequently used to develop an ontology-based information extraction capability. Our approach represents an attempt to combine conventional approaches to information extraction with epidemiological perspectives of culture and network-based approaches to cultural analysis. The approach can be used, we suggest, to support the development of models providing a better understanding of the cognitive characteristics of particular cultural groups.

  19. Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts

    Energy Technology Data Exchange (ETDEWEB)

    Steponkus, P.L.

    1991-01-01

    This project focuses on lesions in the plasma membrane of protoplasts that occur during freezing to temperatures below {minus}5{degrees} which result in changes in the semipermeablity of the plasma membrane. This injury, referred to as loss of osmotic responsiveness, is associated with the formation of large, aparticulate domains in the plasma membrane, aparticulate lamellae subtending the plasma membrane, and lamellar-to-hexagonal{sub II} phase transitions in the plasma membrane and subtending lamellar. The goals of this project are to provide a mechanistic understanding of the mechanism by which freeze-induced dehydration effects the formation of aparticulate domains and lamellar-to-hexagonal{sub II} phase transitions and to determine the mechanisms by which cold acclimation and cryoprotectants preclude or diminish these ultrastructural changes. Our working hypothesis is the formation of aparticulate domains and lamellar-to-hexagon{sub II} phase transitions in the plasma membrane and subtending lamellae are manifestations of hydration-dependent bilayer-bilayer interactions.

  20. [14C]-Sucrose uptake by guard cell protoplasts of pisum sativum, argenteum mutant

    International Nuclear Information System (INIS)

    Rohrig, K.; Raschke, K.

    1991-01-01

    Guard cells rely on import for their supply with reduced carbon. The authors tested by silicone oil centrifugation the ability of guard cell protoplasts to accumulated [ 14 C]-sucrose. Uptake rates were corrected after measurement of 14 C-sorbitol and 3 H 2 O spaces. Sucrose uptake followed biphasic kinetics, with a high-affinity component below 1 mM external sucrose (apparent K m 0.8 mM at 25C) and a low-affinity nonsaturable component above. Uptake depended on pH (optimum at pH 5.0). Variations in the concentrations of external KCl, CCCP, and valinomycin indicated that about one-half of the sucrose uptake rate could be related to an electrochemical gradient across the plasmalemma. Total uptake rates measured at 5 mM external sucrose seem to be sufficient to replenish emptied plastids with starch within a few hours

  1. Laser mutagenesis and producing cellulase condition optimization of Trichoderma virid protoplast

    International Nuclear Information System (INIS)

    Chen Shuli; Zhang Qin; Han Jingjing; Lv Jiangtao; Wang Shilong; Yao Side

    2009-01-01

    The protoplast of Trichoderma virid CICC13038 was mutated using Nd:YAG laser of 266 nm light. And a high-cellulase producing strain JG13 was bred by screening with cellulose microcrystalline. Under the condition of 28 degree C, 180 rpm and 72 h of fermentation time, optimal conditions for the celluase ferment by orthogonal experiment were: 2% bran as the carbon source, 1% (NH 4 ) 2 SO 4 as the nitrogen source, 0.5% Tween-80 as a enzyme-promoting agent,and 25 mL of medium volume in a 250 mL bottle. The cellulase activity of the mutant reached 35.68 U/mL, 25.76% higher than that of the original strain under the same conditions. The mutant JG13 has a great potential in industrial production. And it also can be used as the original strain for further mutagenesis to get the strain of higher cellulase activity. (authors)

  2. Characterization of clays used in the fabrication of traditional brazilian ceramic pans: culture and technique

    International Nuclear Information System (INIS)

    Borlini, Monica Castoldi; Aguiar, Mariane Costalonga de; Vieira, Carlos Mauricio Fontes; Monteiro, Sergio Neves

    2009-01-01

    The fabrication process of clay pans in the state of Espirito Santo, southeast of Brazil, is a recognized part of the country's popular culture. In Goiabeiras, a district of the state capital Vitoria, the traditional production of these pans is the source of income for many families. The technique used in these ceramic pans is of indigenous origin, characterized by manual molding, outdoor burning and application of tannin dye. The clay pans are distributed to several Brazilian states and are nowadays conquering the external market. In producing these pans, two types of, yellow and gray, clays are used. The actual source of raw material comes from the deposit of the Mulemba valley, where a concern on the possibility of exhaustion exists. The objective of this study was then to characterize these two types of clays and so contribute to the continuity of traditional clay pan production by knowing the characteristics of the local clays in case of an eventual need for their replacement. Chemical analysis, X-ray diffraction, particle size distribution, plasticity and thermal analysis of the clays were performed. The results showed that the clays are high plasticity kaolinite with considerable amounts of SiO 2 and Al 2 O 3 as well as of alkaline oxides, earth alkaline oxides and Fe 2 O 3 . (author)

  3. Nuclear techniques used in study of biological processes in Sinapis alba culture

    International Nuclear Information System (INIS)

    Giosanu, D.; Fleancu, M.

    2001-01-01

    The aim of the present paper is to study different nuclear techniques, in particular the influence of gamma radiation upon germination, growth and respiration processes in Sinapis alba culture. The dependence of these phenomena on dose of gamma irradiation was studied. Research was done on dry seeds of mustard (Sinapis alba).The doses of gamma irradiation were: 20 krad, 40 krad, 60 krad, 80 krad and 100 krad.The subsequent evolution of the irradiated samples was compared with the evolution of an unirradiated (control) samples. The irradiation was done evenly, in a single phase. The treatment of the dry seeds of mustard with gamma radiation determined a diminution of energy of germination. So, the energy of germination was 57 - 73% in gamma treated batches and 81% in the control batch. Thus, the faculty of germination decreases from 92% (in the control batch) to 83% in the irradiated batches. Growth process (length of roots and hypocotyl) was also studied. For 100 krad gamma irradiation the rate of this process was lower than that of the control batch, both in the first and the four day of irradiation. The inhibition effect manifested on germination and growth processes for gamma treated dry seeds of mustard is determined by the modification in the membrane permeability. The intensity of respiration process in the irradiated lots was lower than that of the control lot. The inhibition effect manifested by respiration process following gamma irradiation could be explained by the enzymatic activity of mustard seeds. (authors)

  4. Methods and Techniques of Sampling, Culturing and Identifying of Subsurface Bacteria

    International Nuclear Information System (INIS)

    Lee, Seung Yeop; Baik, Min Hoon

    2010-11-01

    This report described sampling, culturing and identifying of KURT underground bacteria, which existed as iron-, manganese-, and sulfate-reducing bacteria. The methods of culturing and media preparation were different by bacteria species affecting bacteria growth-rates. It will be possible for the cultured bacteria to be used for various applied experiments and researches in the future

  5. Transient and stable expression of marker genes in cotransformed Petunia protoplasts in relation to X-ray and UV-irradiation

    International Nuclear Information System (INIS)

    Benediktsson, I.; Köhler, F.; Schieder, O.

    1991-01-01

    Irradiation of protoplasts with X-rays or ultraviolet light does not seem to influence the level of transient expression of foreign DNA in Petunia protoplasts, whereas the number of stably transformed colonies is significantly raised. This may indicate that irradiation influences integration and/or the expression of marker genes and does not result in enhanced uptake rates of plasmids into protoplasts and cell nuclei. Co-transformation with plasmids carrying a gene for kanamycin resistance (neomycin phosphotransferase II) and a gene for hygromycin resistance (hygromycin phosphotransferase) revealed that the cotransformation rates were not stimulated by irradiation when measuring expression

  6. Evaluation of the MODS culture technique for the diagnosis of tuberculous meningitis.

    Directory of Open Access Journals (Sweden)

    Maxine Caws

    2007-11-01

    Full Text Available Tuberculous meningitis (TBM is a devastating condition. The rapid instigation of appropraite chemotherapy is vital to reduce morbidity and mortality. However rapid diagnosis remains elusive; smear microscopy has extremely low sensitivity on cerebrospinal fluid (CSF in most laboratories and PCR requires expertise with advanced infrastructure and has sensitivity of only around 60% under optimal conditions. Neither technique allows for the microbiological isolation of M. tuberculosis and subsequent drug susceptibility testing. We evaluated the recently developed microscopic observation drug susceptibility (MODS assay format for speed and accuracy in diagnosing TBM.Two hundred and thirty consecutive CSF samples collected from 156 patients clinically suspected of TBM on presentation at a tertiary referal hospital in Vietnam were enrolled into the study over a five month period and tested by Ziehl-Neelsen (ZN smear, MODS, Mycobacterial growth Indicator tube (MGIT and Lowenstein-Jensen (LJ culture. Sixty-one samples were from patients already on TB therapy for >1day and 19 samples were excluded due to untraceable patient records. One hundred and fifty samples from 137 newly presenting patients remained. Forty-two percent (n = 57/137 of patients were deemed to have TBM by clinical diagnostic and microbiological criteria (excluding MODS. Sensitivity by patient against clinical gold standard for ZN smear, MODS MGIT and LJ were 52.6%, 64.9%, 70.2% and 70.2%, respectively. Specificity of all microbiological techniques was 100%. Positive and negative predictive values for MODS were 100% and 78.7%, respectively for HIV infected patients and 100% and 82.1% for HIV negative patients. The median time to positive was 6 days (interquartile range 5-7, significantly faster than MGIT at 15.5 days (interquartile range 12-24, and LJ at 24 days (interquartile range 18-35 days (P<0.01.We have shown MODS to be a sensitive, rapid technique for the diagnosis of TBM with

  7. Using Gamma Irradiation To Induce New Mutants In Potatoes Cv. Diamant Through Tissue Culture Technique

    International Nuclear Information System (INIS)

    Sharabash, M.T.; Ali, Amina A. M.; Ahmed, F. A.; Afifi, Abd El-Moneim M.

    2004-01-01

    The excess salt, usually NaCl, inhibits potato plant growth and decreases tubers yield. The use of gamma irradiation to induce new mutants in potato cv. Diamant through tissue culture technique was the main task of this study. Sterilized meristemic tips of potato tubers were cultured on aseptic solid MS-medium, pH 5.7, and were incubated at 20 ± 2 d eg C and 16 hrs day length of 3000-Lux light intensity, to produce virus-free plantlets. Micro-propagation started after 6-8 weeks and plantlets were sub-cultured every 3-4 weeks to increase plantlets population. Plantlets were exposed to 0, or 40 Gy, dose rate 27.7 rad / sec., using Co 60 source at the National Center for Research and Radiation Technology, Cairo, Egypt. Irradiated and unirradiated plantlets were transplanted into 60 ml liquid 1/2MS-medium, pH 5.7, and supplemented with 0, 2000 or 4000 ppm NaCl. And, they were incubated for 2 weeks under the same conditions of temperature and light till the new plantlets were grown up. Healthy plantlets were selected, and micro-propagated up to the sixth vegetative generation (M 1 V 6 ), under the same conditions of salinity and incubation conditions Thereafter, the plantlets were transferred to tuberization liquid 1/2MS-medium, supplemented with the same mentioned concentrations of NaCl, to obtain microtubers. The microtubers were collected after 6-8 weeks and preserved at 10 deg C for 3 months approximately, to break the dormancy. Sprouted microtubers were sown to obtain minitubers, and subsequently macrotubers. All cultures were performed in 30-cm pots in a protected greenhouse, and were irrigated with the same concentrations of NaCl. It could be elicited that cv. Diamant is salinity sensitive. This was evidenced by the decrease in the average number of tubers per plant and average fresh weight of tuber under salinity stress up 4000 ppm NaCl, comparing to unsaline control treatment. Potato plants, which still healthy and produced tubers under salinity stress up to

  8. Light and electron microscopic localization of GABAA-receptors on cultured cerebellar granule cells and astrocytes using immunohistochemical techniques

    DEFF Research Database (Denmark)

    Hansen, Gert Helge; Hösli, E; Belhage, B

    1991-01-01

    . At the light microscope level specific staining of GABAA-receptors was localized in various types of neurones in explant cultures of rat cerebellum using the indirect peroxidase-antiperoxidase (PAP) technique, whereas no specific staining was found in astrocytes. At the electron microscope level labeling...

  9. Analysis of hairy root culture of Rauvolfia serpentina using direct analysis in real time mass spectrometric technique.

    Science.gov (United States)

    Madhusudanan, K P; Banerjee, Suchitra; Khanuja, Suman P S; Chattopadhyay, Sunil K

    2008-06-01

    The applicability of a new mass spectrometric technique, DART (direct analysis in real time) has been studied in the analysis of the hairy root culture of Rauvolfia serpentina. The intact hairy roots were analyzed by holding them in the gap between the DART source and the mass spectrometer for measurements. Two nitrogen-containing compounds, vomilenine and reserpine, were characterized from the analysis of the hairy roots almost instantaneously. The confirmation of the structures of the identified compounds was made through their accurate molecular formula determinations. This is the first report of the application of DART technique for the characterization of compounds that are expressed in the hairy root cultures of Rauvolfia serpentina. Moreover, this also constitutes the first report of expression of reserpine in the hairy root culture of Rauvolfia serpentina. Copyright (c) 2008 John Wiley & Sons, Ltd.

  10. Culture.

    Science.gov (United States)

    Smith, Timothy B; Rodríguez, Melanie Domenech; Bernal, Guillermo

    2011-02-01

    This article summarizes the definitions, means, and research of adapting psychotherapy to clients' cultural backgrounds. We begin by reviewing the prevailing definitions of cultural adaptation and providing a clinical example. We present an original meta-analysis of 65 experimental and quasi-experimental studies involving 8,620 participants. The omnibus effect size of d = .46 indicates that treatments specifically adapted for clients of color were moderately more effective with that clientele than traditional treatments. The most effective treatments tended to be those with greater numbers of cultural adaptations. Mental health services targeted to a specific cultural group were several times more effective than those provided to clients from a variety of cultural backgrounds. We recommend a series of research-supported therapeutic practices that account for clients' culture, with culture-specific treatments being more effective than generally culture-sensitive treatments. © 2010 Wiley Periodicals, Inc.

  11. Microbial community composition of the ileum and cecum of broiler chickens as revealed by molecular and culture-based techniques.

    Science.gov (United States)

    Bjerrum, L; Engberg, R M; Leser, T D; Jensen, B B; Finster, K; Pedersen, K

    2006-07-01

    The microbial communities of the ileum and cecum of broiler chickens from a conventional and an organic farm were investigated using conventional culture techniques as well as cloning and sequencing of 16S rRNA genes. Eighty-five percent of the 557 cloned sequences were <97% related to known cultured species. The chicken ileum was dominated by lactobacilli, whereas the cecum harbored a more diverse microbial community. The cecum was dominated by a large group of bacteria with hitherto no close cultured relatives but most closely related to Faecalibacterium prausnitzii. Approximately 49 and 20% of the cecal clones belonged to this cluster in conventional and organic broiler chickens, respectively. We were, however, able to recover a number of these phylotypes by cultivation, and the isolates were shown to be butyric acid producers. The investigation was a descriptive rather than a comparative study of 2 different rearing systems; however, several differences were observed. For instance, Clostridium perfringens was found in significantly higher numbers in the birds from the organic farm compared with the conventional broilers, probably due to the addition of salinomycin to the conventional feed. In the ileum, the abundance of the different Lactobacillus species differed between the 2 broiler types. The culture-based and culture-independent techniques complemented each other well. Strengths and limitations of the different methods are discussed.

  12. The repertory grid technique as method for the study of cultural differences

    NARCIS (Netherlands)

    Tomico Plasencia, O.; Karapanos, E.; Levy, P.D.; Mizutani, N.; Yamanaka, T.

    2009-01-01

    Culture is typically approached in the field of design through generic, cross-domain constructs. In this paper we provide an alternative methodological approach to exploring cross-cultural differences by studying the idiosyncratic views of individuals with regard to existing products. We

  13. The examination of Hevea brasiliensis plants produced by in vitro culture and mutagenesis by DNA fingerprinting techniques

    International Nuclear Information System (INIS)

    Low, F.C.; Atan, S.; Jaafar, H.

    1998-01-01

    Rubber (Hevea brasiliensis) plants derived from anther and ovule culture as well as gamma-irradiated plants were examined by several DNA marker techniques. These include restriction fragment length polymorphisms (RFLPs), random amplified polymorphic DNA (RAPD), sequence tagged microsatellite sites (STMS), DNA amplification fingerprinting (DAF) and amplified fragment length polymorphisms (AFLPs). Compared to control plants produced by vegetative propagation (cutting and budding), plants produced by in vitro culture appeared to have a reduction in the number of rDNA loci. Two RAPD protocols were compared and found to be similar in amplification of the major DNA bands. After confirmation that the RAPD method adopted was reproducible, the technique was applied to the present studies. Eight out of the 60 primers screened were able to elicit polymorphisms between pooled DNA from in vitro culture plants. Variations in DNA patterns were observed between pooled DNA samples of anther-derived plants as well as between anther-derived and ovule-derived plants. Comparisons of RAPD patterns obtained between anther-derived plants exposed to increasing dosages of gamma-irradiation with non irradiated anther-derived plants revealed distinct DNA polymorphisms. The changes in DNA profiles did not appear to be correlated to the dosage of irradiation. Since somaclonal variation was detected, it was difficult to identify changes which were specifically caused by irradiation. Application of the STMS technique to tag micro satellite sequences (GA) n , (TA) n and (TTA) n in the hydroxymethylglutaryl coenzyme A reductase-1 (hmgr-1) gene failed to detect differences between plants derived from anther and ovule culture. Although restriction endonuclease digestions with methylation sensitive enzymes suggested that four in vitro culture plants examined exhibited similar digestion patterns as the controls, a change in cytosine methylation in one anther-derived plant was detected. Examination of

  14. Isolamento e regeneração de protoplastos de Magnaporthe grisea Isolation and regeneration of Magnaporthe grisea protoplasts

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Marchi

    2006-09-01

    Full Text Available Protoplastos são ferramentas biológicas importantes para pesquisas em fungos filamentosos, sendo empregados intensamente em transformação genética. O isolamento de protoplastos de Magnaporthe grisea foi facilitado com Novozym 234, contudo, este complexo enzimático encontra-se indisponível no mercado. Assim, objetivou-se comparar a eficiência de enzimas líticas disponíveis comercialmente na obtenção de protoplastos de M. grisea. Paralelamente, analisaram-se estabilizadores osmóticos, tempos de digestão e freqüência de regeneração. Maior produção de protoplastos foi obtida com o uso simultâneo de Lysing Enzymes e Cellulase Onozuka R-10. O uso de 10 ou 15 mg de cada complexo enzimático, em 3 mL de estabilizador osmótico, resultou em maior liberação de protoplastos. O melhor estabilizador osmótico foi MgSO4 1,2 M / NaH2PO4 0,01 M, pH 5,8, seguido por MgSO4 0,8 M / NaH2PO4 0,01 M, pH 5,8. O isolamento de protoplastos foi monitorado a cada 60 minutos, atingindo o máximo após incubação por 3 a 6 horas. No entanto, maior freqüência de regeneração (19,4% foi registrada para protoplastos obtidos após 3 horas de hidrólise enzimática.Protoplasts are important biological tools in filamentous fungi research. Fungal protoplasts have been extensively used in experiments with genetic transformation. Protoplastization of Magnaporthe grisea was accomplished with Novozym 234, however, this enzymatic complex is no commercially available for purchase. Thus, the efficiency of several other commercial enzymes in M. grisea protoplasts preparation was investigated. At the same time, osmotic buffer, digestion time and regeneration rate were also analyzed. The highest protoplasts production was obtained with Lysing Enzymes plus Cellulase Onozuka R-10. The use of 10 or 15 mg of each enzymatic complex in 3 mL of osmotic buffer was most effective for the protoplasts yields. The best osmotic buffer was MgSO4 1.2 M / NaH2PO4 0.01 M, pH 5

  15. Project on production of mutants by irradiation of in vitro cultured tissues of coconut and banana and their mass propagation by the tissue culture technique

    International Nuclear Information System (INIS)

    Guzman, E.V. de

    1975-01-01

    Fruit pulp tissue, ovary segments with or without ovules and sections from shoot tips of banana were used for studies on growth stimulating or morphogenetic effects of irradiation. Irradiation at 0.1-1.0 kR tended to induce faster callus growth in the otherwise slow-growing cultures. The physical condition and composition of the culture media especially with respect to growth regulators were studied, as were techniques to overcome discoloration of explants, the best choice of plant tissue for explant, and radiation effects on growth and morphogenesis. Due to the difficulty of callus induction with coconut, only the effects of irradiation on embryos cultured in vitro were studied. They were irradiated at various stages of development, i.e. during the early and final stage of liquid culture, and several days after transfer to a solid medium. Adverse effects of irradiation became evident only during the subsequent growth in solid, during the latter stage of which morphological changes were observed. Whereas irradiation of the liquid as well as solid media up to 50 kR had no adverse effect; survival and development became adversely affected at a dose of 1 kR

  16. Comparative conventional- and quantum dot-labelling strategies for LPS binding site detection in Arabidopsis thaliana mesophyll protoplasts

    Directory of Open Access Journals (Sweden)

    Londiwe Siphephise Mgcina

    2015-05-01

    Full Text Available Lipopolysaccharide (LPS from Gram-negative bacteria is recognized as a microbe-associated molecular pattern (MAMP and not only induces an innate immune response in plants, but also stimulates the development of characteristic defense responses. However, identification and characterization of a cell surface LPS-receptor/binding site, as described in mammals, remains elusive in plants. As an amphiphilic, macromolecular lipoglycan, intact LPS potentially contains three MAMP-active regions, represented by the O-polysaccharide chain, the core and the lipid A. Binding site studies with intact labelled LPS were conducted in Arabidopsis thaliana protoplasts and quantified using flow cytometry fluorescence changes. Qdots, which allow non-covalent, hydrophobic labelling were used as a novel strategy in this study and compared to covalent, hydrophilic labelling with Alexa 488. Affinity for LPS-binding sites was clearly demonstrated by concentration-, temperature- and time-dependent increases in protoplast fluorescence following treatment with the labelled LPS. Moreover, this induced fluorescence increase was convincingly reduced following pre-treatment with excess unlabeled LPS, thereby indicating reversibility of LPS binding. Inhibition of the binding process is also reported using endo- and exocytosis inhibitors. Here, we present evidence for the anticipated presence of LPS-specific binding sites in Arabidopsis protoplasts, and furthermore propose Qdots as a more sensitive LPS-labelling strategy in comparison to the conventional Alexa 488 hydrazide label for binding studies.

  17. Effect of microgravity environment on cell wall regeneration, cell divisions, growth, and differentiation of plants from protoplasts (7-IML-1)

    Science.gov (United States)

    Rasmussen, Ole

    1992-01-01

    The primary goal of this project is to investigate if microgravity has any influence on growth and differentiation of protoplasts. Formation of new cell walls on rapeseed protoplasts takes place within the first 24 hours after isolation. Cell division can be observed after 2-4 days and formation of cell aggregates after 5-7 days. Therefore, it is possible during the 7 day IML-1 Mission to investigate if cell wall formation, cell division, and cell differentiation are influenced by microgravity. Protoplasts of rapeseeds and carrot will be prepared shortly before launch and injected into 0.6 ml polyethylene bags. Eight bags are placed in an aluminum block inside the ESA Type 1 container. The containers are placed at 4 C in PTCU's and transferred to orbiter mid-deck. At 4 C all cell processes are slowed down, including cell wall formation. Latest access to the shuttle will be 12 hours before launch. In orbit the containers will be transferred from the PTC box to the 22 C Biorack incubator. The installation of a 1 g centrifuge in Biorack will make it possible to distinguish between effects of near weightlessness and effects caused by cosmic radiation and other space flight factors including vibrations. Parallel control experiments will be carried out on the ground. Other aspects of the experiment are discussed.

  18. Virtual Reality Techniques for Eliciting Empathy and Cultural Awareness: Affective Human-Virtual World Interaction

    OpenAIRE

    Chirino-Klevans, Ivonne

    2017-01-01

    On the average human beings have about 50,000 thoughts every day. If we consider that thoughts influence how we feel there is little doubt that the way we perceive reality will strongly correlate with how we act upon that reality. Let’s contextualize this thinking process within the realm of global business where interacting with individuals from other cultural backgrounds is the norm. Our own perceptions and stereotypes towards those cultural groups will strongly influence how we interact wi...

  19. Effect of Different Concentrations of Growth Regulators on Gardenia jasminoides cv. Veitchii Micropropagation by Tissue Culture Technique

    Directory of Open Access Journals (Sweden)

    G. R. Abdullah

    2003-01-01

    Full Text Available Micropropagation techniques were set up for Gardenia jasminoides c.v. veitchi. Many plantlets were obtained by culturing shoot cuttings in MS nutrient media, 30 g/L Sucrose, 7 g/L Agar Agar, and different concentrations of BAP and IAA. The best concentration was 1mg /L BAP with 0.5 mg/L IAA. This concentration gave the best sprout growth suitable for rooting in primary and secondary culture by reculturing the stuck cutting every 6 weeks and for many times. We also obtained a high rooting percentage up to 98 % of natural rooting in rooting media different from propagation media by reducing mineral salt concentration to half, Sucrose to 20gm/L, and 2gm/L active charcoal, and 1mg/L IAA. Plantlets were transferred to greenhous and subjected for hardening. This technique gave 22 plantlets from one cutting in one year.

  20. A double labeling technique for performing immunocytochemistry and in situ hybridization in virus infected cell cultures and tissues

    International Nuclear Information System (INIS)

    Gendelman, H.E.; Moench, T.R.; Narayan, O.; Griffin, D.E.; Clements, J.E.

    1985-01-01

    This report describes a combined immunocytochemical and in situ hybridization procedure which allows visualization of cellular or viral antigens and viral RNA in the same cell. Cultures infected with visna or measles virus were fixed in periodate-lysine-paraformaldehyde-glutaraldehyde, stained by the avidin-biotin-peroxidase technique using antibodies to viral or cellular proteins and then incubated with radiolabeled specific DNA probes (in situ hybridization). This technique provides a new approach to the study of viral pathogenesis by: (1) identifying the types of cells which are infected in the host and (2) identifying points of blockade in the virus life cycle during persistent infections. (Auth.)

  1. Diagnosis of bacterial vaginosis in a rural setup: Comparison of clinical algorithm, smear scoring and culture by semiquantitative technique

    OpenAIRE

    Rao P; Devi S; Shriyan A; Rajaram M; Jagdishchandra K

    2004-01-01

    This study was undertaken to estimate the prevalence of bacterial vaginosis (BV) and other sexually transmitted infections (STIs) in a rural set up and compare the smear scoring system to that of culture by semiquantitative technique. A total of 505 married women, who were in sexually active age group of 15-44 years, were selected from three different villages. High vaginal swabs, endocervical swabs, vaginal discharge and blood were collected and processed in the laboratory. Overall prevalenc...

  2. Production of Trichoderma strains with pesticide-polyresistance by mutagenesis and protoplast fusion.

    Science.gov (United States)

    Hatvani, Lóránt; Manczinger, László; Kredics, László; Szekeres, András; Antal, Zsuzsanna; Vágvölgyi, Csaba

    2006-01-01

    The sensitivity of two cold-tolerant Trichoderma strains belonging to the species T. harzianum and T. atroviride was determined to a series of pesticides widely used in agriculture. From the 16 pesticides tested, seven fungicides: copper sulfate, carbendazim, mancozeb, tebuconazole, imazalil, captan and thiram inhibited colony growth of the test strains significantly with minimal inhibitory concentrations of 300, 0.4, 50, 100, 100, 100 and 50 microg/ml, respectively. Mutants resistant to carbendazim and tebuconazole were produced from both wild type strains by means of UV-mutagenesis. The cross-resistance capabilities and in vitro antagonistic properties of the mutants were determined. Carbendazim-resistant mutants showed total cross-resistance to benomyl and thiabendazole at a concentration of 20 microg/ml. Intraspecific protoplast fusion was carried out between carbendazim- and tebuconazole-resistant mutants of both parental strains, and putative haploid recombinants with stable resistance to both pesticides were produced in the case of T. atroviride. These pesticide-polyresistant progenies are potential candidates for application in an integrated pest management system.

  3. Mise au point d'une technique de culture in vitro d'embryons immatures de Phaseolus

    Directory of Open Access Journals (Sweden)

    Véronique Schmit

    1997-01-01

    Full Text Available Development of an in vitro culture technique for immature Phaseolus embryos. In the interspecific crosses Phaseolus polyanthus (or P. coccineus (i? x P. vulgaris, the hybrid embryos abort very early. Therefore, it is essentiel to develop an in vitro culture technique that allows the rescue of beau embryos at globular or early heart-shaped stages. After several trials conceming the salts composition, the sugar rate and the amino acid concentration of différent in vitro culture media, a technique has been developed for heart-shaped Phaseolus embryos. This technique consists of two stages. In a first step, embryos are cultivated under darkness until their germination on a medium containing the salts of Gamborg et al. (1968, 400 mg . 1` (5mM - 1 -' NHNO,, 1 mg . 1-' thiamine HCI, 5 mg . l` nicotinic acid, 0.5 mg - l` pyridoxine, 1,000 mg . l` -glutamine, 1,000 mg . l` casein hydrolysate, 100 mg . l` myo-inositol, 0.028 mg . P N6-benzyladenine, 30 g . l` sucrase, and 8 g -1-' DIFCO agar. After germination, the embryos are cultivated under light on a second medium that does not contain any NHNO, complément and is poorer in amino acids (100 mg • 1-' L-glutamine. Developed with six deys old heart-shaped embryos of the P. vulgaris Bico de Ouro (NI 637 variety, this technique has proved its efficiency with other P. vulgaris and P. polyanthus génotypes. It allows an average régénération rate of 30% from the total number of cultivated embryos.

  4. Micropropagation of Pear Rootstock (Pyrus Communis) by using tissue culture technique and gamma irradiation

    International Nuclear Information System (INIS)

    El-Sharnouby, M.E.; ESSAM, E.R.; Ayoub, S.

    2006-01-01

    New growing shoots from healthy pear rootstock (Pyrus communis) trees were taken and sterilized 3 times in dipping water. Explants were subjected to antioxidant treatment, different media, different additives and different BAP and NAA concentrations. The obtained results showed that Murashig-Skoog (MS) supplemented with 1 mg/l BA was better than Gamborg medium. Adding antioxidant solution and adenine sulphate to the culture medium was preferred for maximizing explants development. Exposing the explants to gamma irradiation at different doses decreased tissue culture parameters with increasing gamma doses. However, the low dose of gamma rays (1 Krad) significantly increased the number of shoots than other gamma treatments. Adding of BAP at 2 mg/l to the culture medium increased number and length of shoots. However, addition of 1 mg/l NAA to the rooting medium led to increase the root formation

  5. Nuclear techniques for the analysis and dating of cultural heritage with the tandetron accelerator at the CEDAD

    Directory of Open Access Journals (Sweden)

    Lucio Calcagnile

    2014-12-01

    Full Text Available The Accelerator Mass Spectrometry technique for measuring carbon isotopes and dating artifacts, together with nuclear techniques of ion beam analysis are widely used in the field of cultural heritage owing to the great advantage of their being non-destructive. In Italy, CEDAD - CEntro Di DAtazione e Diagnostica of the University of Salento, with its tandetron accelerator of 3MV, has played a major role for over ten years, in studying materials of cultural assets requiring the determination of their elemental composition and determination of absolute chronology using radiocarbon. This study describes the Accelerator Mass Spectrometry technique used with the accelerator at CEDAD to determine age by means of radiocarbon and PIXE-PIGE techniques, and to determine elements, even in traces, present in materials. Some case studies carried out at CEDAD are reported, including those on the Riace Bronzes and Capitoline Wolf. The latter has been definitively dated to the Middle Ages, 17 centuries later than the previously attributed dating by historians. In addition, an important technical innovation is described, achieved within the framework of the IT@CHA Project enabling organic materials to be dated using only 10 μg of carbon.

  6. A cell culture technique for human epiretinal membranes to describe cell behavior and membrane contraction in vitro.

    Science.gov (United States)

    Wertheimer, Christian; Eibl-Lindner, Kirsten H; Compera, Denise; Kueres, Alexander; Wolf, Armin; Docheva, Denitsa; Priglinger, Siegfried G; Priglinger, Claudia; Schumann, Ricarda G

    2017-11-01

    To introduce a human cell culture technique for investigating in-vitro behavior of primary epiretinal cells and membrane contraction of fibrocellular tissue surgically removed from eyes with idiopathic macular pucker. Human epiretinal membranes were harvested from ten eyes with idiopathic macular pucker during standard vitrectomy. Specimens were fixed on cell culture plastic using small entomological pins to apply horizontal stress to the tissue, and then transferred to standard cell culture conditions. Cell behavior of 400 epiretinal cells from 10 epiretinal membranes was observed in time-lapse microscopy and analyzed in terms of cell migration, cell velocity, and membrane contraction. Immunocytochemistry was performed for cell type-specific antigens. Cell specific differences in migration behavior were observed comprising two phenotypes: (PT1) epiretinal cells moving fast, less directly, with small round phenotype and (PT2) epiretinal cells moving slowly, directly, with elongated large phenotype. No mitosis, no outgrowth and no migration onto the plastic were seen. Horizontal contraction measurements showed variation between specimens. Masses of epiretinal cells with a myofibroblast-like phenotype expressed cytoplasmatic α-SMA stress fibers and correlated with cell behavior characteristics (PT2). Fast moving epiretinal cells (PT1) were identified as microglia by immunostaining. This in-vitro technique using traction application allows for culturing surgically removed epiretinal membranes from eyes with idiopathic macular pucker, demonstrating cell behavior and membrane contraction of primary human epiretinal cells. Our findings emphasize the abundance of myofibroblasts, the presence of microglia and specific differences of cell behavior in these membranes. This technique has the potential to improve the understanding of pathologies at the vitreomacular interface and might be helpful in establishing anti-fibrotic treatment strategies.

  7. Improvement of potato tolerance to salinity using tissue culture techniques and irradiation with in vitro selection

    International Nuclear Information System (INIS)

    Al-Safadi, B.; Arabi, M. I. E.

    2005-06-01

    A mutation breeding program was conducted to improve potato (Solanum tuberosum) tolerance to salinity. In vitro cultured explants from potato cvs. Draga, Diamant, Spunta were irradiated with gamma doses 25, 30, and 35 Gy.Growing shoots were cut and re-cultured every 2 weeks until the 4th generation (MV 4 ) to make sure no chimeral tissues still existed in the mutant material. Plantlets were subsequently propagated to obtain enough explants for in vitro selection pressure. Around 3000 plantlets from the three cultivars were subjected to selection pressure. MV 4 explants were cultured on MS medium supplemented with the NaCl in varying concentrations ranging between 50 to 200 mM. Surviving plantlets were propagated and re-cultured on a similar medium to insure their tolerance to salinity. Tolerant plantlets were acclimatized and transferred to pots and grown under glasshouse conditions. Plants were later subjected to another selection pressure, by irrigating them using water containing NaCl in concentrations ranging between 50-250 mM in addition to controls irrigated with normal water. Cultivar Spunta produced the highest number of tolerant plants. Four plants of Spunta appeared to be tolerant to salinity whereas only one plant from Diamant and was tolerant and no plants from cultivar Draga were tolerant. Mutant plants varied in number of produced minitubers from 8 - 14. Also, weight of these minitubers varied from less than 1 to 31 grams. (author)

  8. Translation of questionnaires into Arabic in cross-cultural research: techniques and equivalence issues.

    Science.gov (United States)

    Khalaila, Rabia

    2013-10-01

    To describe the translation process of nursing instruments into Arabic and discuss the equivalence issues arising from this process. Review of the literature. The Arabic language is essentially three different languages: Classical Arabic; Modern Standard Arabic (fuS-Ha or MSA); and colloquial Arabic (Lahja A'mmeya), which is itself divided into five different regional Arabic dialects. The Arabic fuS-Ha language is the dialect most widely used in the translation of instruments into Arabic. The literature reveals that only a few studies focused on the linguistic issues in the translation of instruments into Arabic. Brislin's back-translation emerged as the most common method widely used by researchers in studies with Arabic-speaking subjects, but not the perfect one. Linguistic issues in nursing research have not been sufficiently described and discussed in the context of Arabic language and culture. Although there is no standard guideline for instrument translation, the combined translation model is the most recommended procedure to use in cross-cultural research. Linguistic differences between the source culture and the target Arabic culture should be taken into account. Finally, we recommend the use of the fuS-Ha dialect and trilingual translators in the translation of nursing instruments into Arabic.

  9. caractérisation des itinéraires techniques de la culture de courgette ...

    African Journals Online (AJOL)

    USER

    L'implantation à contre temps de la culture, son irrigation et sa protection phytosanitaire aléatoires, et une ... study is the diagnosis of the approaches used in managing zucchini (Cucurbita pepo L.) cultivation in the PIA of .... précédent cultivé par le cinquième agriculteur de ..... alternative for machine-harvested Cucumbers.

  10. Isolation of Mycobacterium chelonei with the lysis-centrifugation blood culture technique.

    OpenAIRE

    Fojtasek, M F; Kelly, M T

    1982-01-01

    Mycobacterium chelonei was isolated from a patient by the lysis-centrifugation and the conventional two-bottle blood culture methods. The lysis-centrifugation method was significantly more sensitive and rapid than the conventional method in detecting and isolating this organism; quantitations done by this method were useful for monitoring response to therapy.

  11. Writing for Immediacy: Narrative Writing as a Teaching Technique in Undergraduate Cultural Geography

    Science.gov (United States)

    Summerby-Murray, Robert

    2010-01-01

    Narrative inquiry is an innovative means of encouraging students to internalize concepts, reflect on experiences or create applications for theoretical ideas. The use of first-person creative writing in a second-year cultural geography course prompted initial scepticism from students but eventually highlighted their constructivist engagement with…

  12. Human nasal turbinates as a viable source of respiratory epithelial cells using co-culture system versus dispase-dissociation technique.

    Science.gov (United States)

    Noruddin, Nur Adelina Ahmad; Saim, Aminuddin B; Chua, Kien Hui; Idrus, Ruszymah

    2007-12-01

    To compare a co-culture system with a conventional dispase-dissociation method for obtaining functional human respiratory epithelial cells from the nasal turbinates for tissue engineering application. Human respiratory epithelial cells were serially passaged using a co-culture system and a conventional dispase-dissociation technique. The growth kinetics and gene expression levels of the cultured respiratory epithelial cells were compared. Four genes were investigated, namely cytokeratin-18, a marker for ciliated and secretory epithelial cells; cytokeratin-14, a marker for basal epithelial cells; MKI67, a proliferation marker; and MUC5B, a marker for mucin secretion. Immunocytochemical analysis was performed using monoclonal antibodies against the high molecular-weight cytokeratin 34 beta E12, cytokeratin 18, and MUC5A to investigate the protein expression from cultured respiratory epithelial cells. Respiratory epithelial cells cultured using both methods maintained polygonal morphology throughout the passages. At passage 1, co-cultured respiratory epithelial showed a 2.6-times higher growth rate compared to conventional dispase dissociation technique, and 7.8 times higher at passage 2. Better basal gene expression was observed by co-cultured respiratory epithelial cells compared to dispase dissociated cells. Immunocytochemical analyses were positive for the respiratory epithelial cells cultured using both techniques. Co-culture system produced superior quality of cultured human respiratory epithelial cells from the nasal turbinates as compared to dispase dissociation technique.

  13. Quality management techniques in public sector organizations: the role of managerial autonomy and organizational culture

    OpenAIRE

    Demuzere, Sara; Verhoest, Koen; Bouckaert, Geert

    2008-01-01

    According to NPM, more effective and efficient public services can be delivered if public sector organizations are granted more managerial autonomy and if they use various (private sector) management techniques. To date, only very limited attention has been paid to the link between managerial autonomy and the use of management techniques. With this study, we seek to fill this gap in the extensive NPM literature by examining the extent to which managerial autonomy has an effect on the use of m...

  14. Use of radiation for inducing mutants in potatoes through tissue culture technique

    International Nuclear Information System (INIS)

    Sharabash, M.T.

    1997-01-01

    Meristem-tips obtained from sprouts of potato tubers, cv. 'Diamant' (Solanum tuberosum) were cultured on MS-medium and multiplied into plantlets through micropropagation. After 2-3 weeks, the micropropagated plantlets had 5-6 nodes each. The plantlets were irradiated with 20 to 40 Gy gamma rays at 27.7 rad/sec. Irradiated plantlets were cut into single nodes and cultured on MS-medium supplemented with 2 g NaCl. Salt resistant plantlets were transferred to MS-liquid medium supplemented with 2g NaCl/l 5.2, and microtubers were collected after 6 weeks. Minitubers were produced under the same level of salinity. (author). 2 tabs

  15. Impact of starter cultures and fermentation techniques on the volatile aroma and sensory profile of chocolate

    DEFF Research Database (Denmark)

    Crafack, Michael; Keul, Hanna; Eskildsen, Carl Emil Aae

    2014-01-01

    cultures on the formation of flavour precursors, composition of volatile aroma compounds and sensory profile was investigated in cocoa inoculated with cultures encompassing a highly aromatic strain of Pichia kluyveri or a pectinolytic strain of Kluyveromyces marxianus, and compared to commercially...... fermented heap and tray cocoa. Although only minor differences in the concentration of free amino acids and reducing sugars was measured, identification and quantification by dynamic headspace gas chromatography-mass spectrometry (HS/GC-MS) revealed pronounced differences in the composition of volatiles...... in roasted cocoa liquors and finished chocolates. 19 of the 56 volatile compounds identified in the chocolates were found in significantly higher amounts in the tray fermented sample, whilst significantly higher amounts of 2-methoxyphenol was measured in the two inoculated chocolates. The P. kluyveri...

  16. Use of radiation for inducing mutants in potatoes through tissue culture technique

    Energy Technology Data Exchange (ETDEWEB)

    Sharabash, M T [National Centre for Radiation Research and Technology, Cairo (Egypt)

    1997-07-01

    Meristem-tips obtained from sprouts of potato tubers, cv. `Diamant` (Solanum tuberosum) were cultured on MS-medium and multiplied into plantlets through micropropagation. After 2-3 weeks, the micropropagated plantlets had 5-6 nodes each. The plantlets were irradiated with 20 to 40 Gy gamma rays at 27.7 rad/sec. Irradiated plantlets were cut into single nodes and cultured on MS-medium supplemented with 2 g NaCl. Salt resistant plantlets were transferred to MS-liquid medium supplemented with 2g NaCl/l 5.2, and microtubers were collected after 6 weeks. Minitubers were produced under the same level of salinity. (author). 2 tabs.

  17. Efficiency of anther culture technique in the production of wheat double haploids

    Directory of Open Access Journals (Sweden)

    Kondić-Špika Ankica Đ.

    2008-01-01

    Full Text Available The objective of the study was to investigate efficiency of anther culture in the production of spontaneous double haploids from randomly selected heterozygous genotypes of wheat (Triticum aestivum L.. Anthers of 20 F1 wheat combinations were grown in vitro on a modified Potato-2 medium. All of the examined genotypes have shown the ability to produce pollen calluses as well as to regenerate green plants. On average for the whole experiment material, 47.2 calluses were produced per 100 cultured anthers. The green plant regeneration ranged from 0.8 to 13.4 green plants per spike, with an overall mean of 5.8. From the total of 582 regenerated green plants, 47.9% (279 were spontaneous double haploids. The final average yield from the study was 2.8 double haploids per spike.

  18. Next-generation sequencing and culture-based techniques offer complementary insights into fungi and prokaryotes in beach sands.

    Science.gov (United States)

    Romão, Daniela; Staley, Christopher; Ferreira, Filipa; Rodrigues, Raquel; Sabino, Raquel; Veríssimo, Cristina; Wang, Ping; Sadowsky, Michael; Brandão, João

    2017-06-15

    A next-generation sequencing (NGS) approach, in conjunction with culture-based methods, was used to examine fungal and prokaryotic communities for the presence of potential pathogens in beach sands throughout Portugal. Culture-based fungal enumeration revealed low and variable concentrations of the species targeted (yeasts and dermatophytes), which were underrepresented in the community characterized by NGS targeting the ITS1 region. Conversely, NGS indicated that the potentially pathogenic species Purpureocillium liliacinum comprised nearly the entire fungal community. Culturable fecal indicator bacterial concentrations were low throughout the study and unrelated to communities characterized by NGS. Notably, the prokaryotic communities characterized revealed a considerable abundance of archaea. Results highlight differences in communities between methods in beach sand monitoring but indicate the techniques offer complementary insights. Thus, there is a need to leverage culture-based methods with NGS methods, using a toolbox approach, to determine appropriate targets and metrics for beach sand monitoring to adequately protect public health. Copyright © 2017. Published by Elsevier Ltd.

  19. Production of monozygotic twin calves using the blastomere separation technique and Well of the Well culture system.

    Science.gov (United States)

    Tagawa, M; Matoba, S; Narita, M; Saito, N; Nagai, T; Imai, K

    2008-03-15

    The present study was conducted to establish a simple and efficient method of producing monozygotic twin calves using the blastomere separation technique. To produce monozygotic twin embryos from zona-free two- and eight-cell embryos, blastomeres were separated mechanically by pipetting to form two demi-embryos; each single blastomere from the two-cell embryo and tetra-blastomeres from the eight-cell embryo were cultured in vitro using the Well of the Well culture system (WOW). This culture system supported the successful arrangement of blastomeres, resulting in their subsequent aggregation to form a demi-embryo developing to the blastocyst stage without a zona pellucida. There was no significant difference in the development to the blastocyst stage between blastomeres separated from eight-cell (72.0%) and two-cell (62.0%) embryos. The production rates of the monozygotic pair blastocysts and transferable paired blastocysts for demi-embryos obtained from eight-cell embryos (64.0 and 45.0%, respectively) were higher than those for demi-embryos obtained from two-cell embryos (49.0 and 31.0%, PWOW culture system, yielded viable monozygotic demi-embryos, resulting in high rates of pregnancy and twinning rates after embryo transfer.

  20. A Rapid Culture Technique Produces Functional Dendritic-Like Cells from Human Acute Myeloid Leukemia Cell Lines

    Directory of Open Access Journals (Sweden)

    Jian Ning

    2011-01-01

    Full Text Available Most anti-cancer immunotherapeutic strategies involving dendritic cells (DC as vaccines rely upon the adoptive transfer of DC loaded with exogenous tumour-peptides. This study utilized human acute myeloid leukemia (AML cells as progenitors from which functional dendritic-like antigen presenting cells (DLC were generated, that constitutively express tumour antigens for recognition by CD8+ T cells. DLC were generated from AML cell lines KG-1 and MUTZ-3 using rapid culture techniques and appropriate cytokines. DLC were evaluated for their cell-surface phenotype, antigen uptake and ability to stimulate allogeneic responder cell proliferation, and production of IFN-γ; compared with DC derived from normal human PBMC donors. KG-1 and MUTZ-3 DLC increased expression of CD80, CD83, CD86, and HLA-DR, and MUTZ-3 DLC downregulated CD14 and expressed CD1a. Importantly, both KG-1 and MUTZ-3-derived DLC promoted proliferation of allogeneic responder cells more efficiently than unmodified cells; neither cells incorporated FITC-labeled dextran, but both stimulated IFN-γ production from responding allogeneic CD8+ T cells. Control DC produced from PBMC using the FastDC culture also expressed high levels of critical cell surface ligands and demonstrated good APC function. This paper indicates that functional DLC can be cultured from the AML cell lines KG-1 and MUTZ-3, and FastDC culture generates functional KG-1 DLC.

  1. A shell-less chick embryo culturing technique, reproduced successfully under local circumstances

    International Nuclear Information System (INIS)

    Zareen, N.; Khan, Y.

    2008-01-01

    The goal of this project was to demonstrate shell-less chick embryo culturing as a potential experimental model in the field of developmental anatomy. Freshly laid, fertilized chicken eggs of Egyptian Fayoumi breed were obtained from Poultry Research Institute Punjab, Rawalpindi. The fertilized chicken eggs were preincubated for 33 hours under standard conditions of 37.5 degree C and 65-75% humidity, to bring them to stage 9 (29-33 hours embryo, 7 somites) of Hamburger and Hamilton staging system. After this period, the eggs were taken out of the incubator, placed horizontally, wiped with 70% ethanol and permitted to air-dry for 10 minutes to reduce contamination from the egg surface and also to ensure that the embryo was properly positioned. The eggs contents were then transferred into the culture containers by cracking the undersides against an edge. The formation and growth of the embryonic membranes, the central nervous system - beginning from the vesicle stage, the circulatory system - including the heart, the eyes, beak, limbs, skin, feathers, wings and folding of the body were directly observed. Repeated successful culturing was attempted, tracing the developmental process of the embryo upto the 15th day of embryonic life at least after which the survivability period varied in different embryo cultures. The most advanced age reached in this project was day 19 of the embryonic life, which in researchers understanding is the latest developmental stage in shellless environment described as yet. The normal hatching time of this breed is 21-22 days. The size of these embryos was smaller as compared to the embryos of the same age that carried out their development inside their shells. (author)

  2. Improvement of potato tolerance to salinity using tissue culture techniques and irradiation with in vitro selection

    International Nuclear Information System (INIS)

    Al-Safadi, B.; Arabi, M. I. E.

    2006-01-01

    A mutation breeding program was conducted to improve potato (Solanum tuberosum) tolerance to salinity. In vitro cultured explants from potato cvs. Draga, Diamant, Spunta were irradiated with gamma doses 25, 30, and 35 Gy. Mutants were isolated to get rid of chimeral tissues and subsequently propagated for in vitro and pot selection pressure. Cultivar Sponta produced the highest number of tolerant plants (4) and only one plant was obtained from Diamant. (authors)

  3. Analytical techniques for characterization of raw materials in cell culture media

    Directory of Open Access Journals (Sweden)

    Sharma Chandana

    2011-11-01

    Full Text Available Abstract Raw materials are a critical part of any cell culture medium; therefore, it is of utmost importance to understand and characterize them for high-quality product. The raw material characterization (RMC program at SAFC focuses on individual screening of raw materials both analytically and biologically. The goal of the program is to develop the best-in-class knowledge base of the raw materials used in SAFC’s media formulations and their impact on performance of products.

  4. Radiation induced variation in potato for tolerance to salinity using tissue culture technique

    International Nuclear Information System (INIS)

    Sharabash, M.T.

    2001-01-01

    Meristem-tips of potato (Solanum tuberosum) cv. 'Diamant', obtained from tuber sprouts, were cultured on MS medium, and multiplied into plantlets through micropropagation. To induce variation for salt tolerance, the obtained plantlets were irradiated with 0, 20, and 40 Gy gamma rays at 27.7 rad/sec. Irradiated plantlets were cut into single nodes and cultured on MS medium, supplemented with 2000 and 4000 ppm NaCI. Salt tolerant plantlets were transferred for tuberization on MS liquid medium supplemented with the same concentration of NaCI. Micro-tubers, collected after 6 weeks of culture, had fresh weight between 0.03 to 0.3 g. Mini-tubers were obtained by planting micro-tubers in 25 cm pots under insect proof greenhouse. Mini-tuber number per plant ranged from 3 to 6, and the mini-tuber weight ranged from 0.5-3.0 g, depending upon the treatment. Further studies are in progress to produce conventional tubers under salinity stress from the promising variants, specially those tolerant to 4000 ppm, and to assure the stability of the obtained variants. (author)

  5. Diagnosis of bacterial vaginosis in a rural setup: comparison of clinical algorithm, smear scoring and culture by semiquantitative technique.

    Science.gov (United States)

    Rao, P S; Devi, S; Shriyan, A; Rajaram, M; Jagdishchandra, K

    2004-01-01

    This study was undertaken to estimate the prevalence of bacterial vaginosis (BV) and other sexually transmitted infections (STIs) in a rural set up and compare the smear scoring system to that of culture by semiquantitative technique. A total of 505 married women, who were in sexually active age group of 15-44 years, were selected from three different villages. High vaginal swabs, endocervical swabs, vaginal discharge and blood were collected and processed in the laboratory. Overall prevalence of 29% reproductive tract infection was detected. Endogenous infection was commonly observed (27.92%), and very low prevalence of STIs (Trichomonas 1.18%, Syphilis 0%, Gonorrhea 0%) was detected. Diagnosis of BV was possible in 104 (20.5%) women by smear alone and 88 (17.42%) women by semiquantitative culture.

  6. Diagnosis of bacterial vaginosis in a rural setup: Comparison of clinical algorithm, smear scoring and culture by semiquantitative technique

    Directory of Open Access Journals (Sweden)

    Rao P

    2004-01-01

    Full Text Available This study was undertaken to estimate the prevalence of bacterial vaginosis (BV and other sexually transmitted infections (STIs in a rural set up and compare the smear scoring system to that of culture by semiquantitative technique. A total of 505 married women, who were in sexually active age group of 15-44 years, were selected from three different villages. High vaginal swabs, endocervical swabs, vaginal discharge and blood were collected and processed in the laboratory. Overall prevalence of 29% reproductive tract infection was detected. Endogenous infection was commonly observed (27.92%, and very low prevalence of STIs (Trichomonas 1.18%, Syphilis 0%, Gonorrhea 0% was detected. Diagnosis of BV was possible in 104 (20.5% women by smear alone and 88 (17.42% women by semiquantitative culture.

  7. Conditions de sélection in vitro de cals issus des disques foliaires et des protoplastes de Pelargonium tolérant plus la sécheresse

    Directory of Open Access Journals (Sweden)

    M. MOKHTARI

    2015-01-01

    Full Text Available To induce drought resistance, callus from leaf discs and protoplast of Pelargonium x hortorum (‘Deep Salmon’ and ‘Panaché Sud’ were grown in vitro in osmoticum rich medium. Percent survival of the callus varied with growth medium, variety and concentration of osmoticum. Compared to protoplasts, leaf discs were simple to handle. However, protoplasts growing requires enzymatic additives and delicate procedures. The protoplasts viability was 86% for ‘Deep Salmon’ and 90% for ‘Panaché Sud’. The yield was 5.67 x 106 protoplasts / g FM for ‘Deep Salmon’ and 11.35 x 106 for ‘Panaché Sud’. The callus from leaf discs of the variety Deep Salmon survived a maximum concentration of 0.5 M sucrose and 0.27 M mannitol or sorbitol. A dose of 0.6 M sucrose was the threshold limit for the survival of 12.5% ​​protoplasts with a division ratio of 2% for ‘Deep Salmon’ and 18.7% of protoplasts with a division ratio of 3.2% for ‘Panaché Sud’. For the mannitol, the maximum limit was 0.6 M for a 13.5% viability of protoplasts with a division ratio 3.6% for ‘Deep Salmon’ and 16.1% of protoplasts with a division factor 3 % respectively for ‘Panaché Sud’. The 20% PEG allowed the survival of 21.1% protoplast and a division rate of 0.2% in ‘Deep Salmon’, but it has totally inhibited protoplast division of ‘Panaché Sud’, even at 5%.

  8. Socio-cultural inhibitors to use of modern contraceptive techniques in rural Uganda: a qualitative study

    Science.gov (United States)

    Kabagenyi, Allen; Reid, Alice; Ntozi, James; Atuyambe, Lynn

    2016-01-01

    Introduction Family planning is one of the cost-effective strategies in reducing maternal and child morbidity and mortality rates. Yet in Uganda, the contraceptive prevalence rate is only 30% among married women in conjunction with a persistently high fertility rate of 6.2 children per woman. These demographic indicators have contributed to a high population growth rate of over 3.2% annually. This study examines the role of socio-cultural inhibitions in the use of modern contraceptives in rural Uganda. Methods This was a qualitative study conducted in 2012 among men aged 15-64 and women aged 15-49 in the districts of Mpigi and Bugiri in rural Uganda. Eighteen selected focus group discussions (FGDs), each internally homogeneous, and eight in-depth interviews (IDIs) were conducted among men and women. Data were collected on sociocultural beliefs and practices, barriers to modern contraceptive use and perceptions of and attitudes to contraceptive use. All interviews were tape recoded, translated and transcribed verbatim. All the transcripts were coded, prearranged into categories and later analyzed using a latent content analysis approach, with support of ATLAS.ti qualitative software. Suitable quotations were used to provide in-depth explanations of the findings. Results Three themes central in hindering the uptake of modern contraceptives emerged: (i) persistence of socio-cultural beliefs and practices promoting births (such as polygamy, extending family lineage, replacement of the dead, gender-based violence, power relations and twin myths). (ii) Continued reliance on traditional family planning practices and (iii) misconceptions and fears about modern contraception. Conclusion Sociocultural expectations and values attached to marriage, women and child bearing remain an impediment to using family planning methods. The study suggests a need to eradicate the cultural beliefs and practices that hinder people from using contraceptives, as well as a need to scale

  9. Socio-cultural inhibitors to use of modern contraceptive techniques in rural Uganda: a qualitative study.

    Science.gov (United States)

    Kabagenyi, Allen; Reid, Alice; Ntozi, James; Atuyambe, Lynn

    2016-01-01

    Family planning is one of the cost-effective strategies in reducing maternal and child morbidity and mortality rates. Yet in Uganda, the contraceptive prevalence rate is only 30% among married women in conjunction with a persistently high fertility rate of 6.2 children per woman. These demographic indicators have contributed to a high population growth rate of over 3.2% annually. This study examines the role of socio-cultural inhibitions in the use of modern contraceptives in rural Uganda. This was a qualitative study conducted in 2012 among men aged 15-64 and women aged 15-49 in the districts of Mpigi and Bugiri in rural Uganda. Eighteen selected focus group discussions (FGDs), each internally homogeneous, and eight in-depth interviews (IDIs) were conducted among men and women. Data were collected on sociocultural beliefs and practices, barriers to modern contraceptive use and perceptions of and attitudes to contraceptive use. All interviews were tape recoded, translated and transcribed verbatim. All the transcripts were coded, prearranged into categories and later analyzed using a latent content analysis approach, with support of ATLAS.ti qualitative software. Suitable quotations were used to provide in-depth explanations of the findings. Three themes central in hindering the uptake of modern contraceptives emerged: (i) persistence of socio-cultural beliefs and practices promoting births (such as polygamy, extending family lineage, replacement of the dead, gender-based violence, power relations and twin myths). (ii) Continued reliance on traditional family planning practices and (iii) misconceptions and fears about modern contraception. Sociocultural expectations and values attached to marriage, women and child bearing remain an impediment to using family planning methods. The study suggests a need to eradicate the cultural beliefs and practices that hinder people from using contraceptives, as well as a need to scale-up family planning services and sensitization

  10. Microbial community composition of the ileum and cecum of broiler chickens as revealed by molecular and culture-based techniques

    DEFF Research Database (Denmark)

    Friis-Holm, Lotte Bjerrum; Engberg, R.M.; Leser, T.D.

    2006-01-01

    The microbial communities of the ileum and cecum of broiler chickens from a conventional and an organic farm were investigated using conventional culture techniques as well as cloning and sequencing of 16S rRNA genes. Eighty-five percent of the 557 cloned sequences were ...% of the cecal clones belonged to this cluster in conventional and organic broiler chickens, respectively. We were, however, able to recover a number of these phylotypes by cultivation, and the isolates were shown to be butyric acid producers. The investigation was a descriptive rather than a comparative study...

  11. Seafood Wars: Reviving a Tired Sustainability Education Program with Pop Culture Techniques

    Science.gov (United States)

    Peart, L. W.

    2016-02-01

    Texas State Aquarium revived its sustainable seafood education program by embedding expert speakers into the pop culture chef competition. Chefs are nominated by diners and vetted by Aquarium staff. Seafood selections are made in consultation with fishery experts and sustainability partners including Gulf United for Lasting Fisheries. Through these efforts, the Seafood Wars audience has expanded from the over-40 set to college and graduate students, families, and adults of all ages. Surveyed participants at these sell-out events are 100% as, or more likely to purchase and consume featured sustainable selections.

  12. A Quantitative Needs Assessment Technique for Cross-Cultural Work Adjustment Training.

    Science.gov (United States)

    Selmer, Lyn

    2000-01-01

    A study of 67 Swedish expatriate bosses and 104 local Hong Kong middle managers tested a quantitative needs assessment technique measuring work values. Two-thirds of middle managers' work values were not correctly estimated by their bosses, especially instrumental values (pay, benefits, security, working hours and conditions), indicating a need…

  13. Simulation of limiting dilution technique in determination of immunocompetent cells frequency in irradiated cell cultures

    International Nuclear Information System (INIS)

    Martini Filho, R.J.; Barlette, V.E.; Goes, E.G.; Covas, D.T.; Orellana, M.

    2001-01-01

    Limiting dilution techniques (LDA) dose-response data have been used to detect immunocompetent T-Cells in microcultures. In this work, LDA frequencies estimates was obtained using χ2 minimization for irradiated cells in a range of 500 to 1,500 cGy. (author)

  14. Production of virus-free orchid Cymbidium aloifolium (L.) Sw. by various tissue culture techniques.

    Science.gov (United States)

    Pradhan, Shreeti; Regmi, Tripti; Ranjit, Mukunda; Pant, Bijaya

    2016-10-01

    Orchids are affected by many viruses resulting in poor growth, yield and quality, and an overall decline in population. Cymbidium mosaic virus (CymMV) is one of the common orchid viruses found in Cymbidium species but it infects different orchid genera. In this study Cymbidium aloifolium was propagated in vitro using MS medium at different strength (1.0, ½, and ¼) with or without 0.5 mg/l BAP (6-benzylaminopurine) and 0.5 mg/l NAA (Naphthalene acetic acid). To provide disease-free planting material, source plant for in vitro propagation needs to be screened for pathogenic viruses. In the present study, in vivo -grown source (mother) plants and tissue culture-derived plants of C. aloifolium were tested for CymMV virus using Double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA). All the tissue cultured plants were found to be 100% virus-free whereas the in vivo grown source plants were highly affected by CymMV virus (83.33%). The virus-free in vitro plantlets were multiplied in large scale and then acclimatized on earthen pot containing a mixture of cocopeat, litter and clay in the ratio of 3:2:1. Eighty five percent of acclimatized plantlets survived making this method an efficient mass production system for high quality virus-free C. aloifolium for commercial floriculture and germplasm preservation.

  15. Production of virus-free orchid Cymbidium aloifolium (L. Sw. by various tissue culture techniques

    Directory of Open Access Journals (Sweden)

    Shreeti Pradhan

    2016-10-01

    Full Text Available Orchids are affected by many viruses resulting in poor growth, yield and quality, and an overall decline in population. Cymbidium mosaic virus (CymMV is one of the common orchid viruses found in Cymbidium species but it infects different orchid genera. In this study Cymbidium aloifolium was propagated in vitro using MS medium at different strength (1.0, ½, and ¼ with or without 0.5 mg/l BAP (6-benzylaminopurine and 0.5 mg/l NAA (Naphthalene acetic acid. To provide disease-free planting material, source plant for in vitro propagation needs to be screened for pathogenic viruses. In the present study, in vivo-grown source (mother plants and tissue culture-derived plants of C. aloifolium were tested for CymMV virus using Double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA. All the tissue cultured plants were found to be 100% virus-free whereas the in vivo grown source plants were highly affected by CymMV virus (83.33%. The virus-free in vitro plantlets were multiplied in large scale and then acclimatized on earthen pot containing a mixture of cocopeat, litter and clay in the ratio of 3:2:1. Eighty five percent of acclimatized plantlets survived making this method an efficient mass production system for high quality virus-free C. aloifolium for commercial floriculture and germplasm preservation. Keywords: Biological sciences, Plant biology

  16. Uncapped mRNA introduced into tobacco protoplasts can be imported into the nucleus and is trapped by leptomycin B.

    Science.gov (United States)

    Stuger, Rogier; Forreiter, Christoph

    2004-08-01

    The mechanism of nuclear export of RNAs in yeast and animal cells is rapidly being uncovered, but RNA export in plants has received little attention. We introduced capped and uncapped fluorescent mRNAs into tobacco (Nicotiana plumbaginifolia) protoplasts and studied their cellular localization. Following insertion, capped transcripts were found in the cytoplasm, while uncapped messengers transiently appeared in the nucleus in about one-quarter to one-third of the cells. These mRNAs were trapped by the nuclear export-inhibiting drug leptomycin B, pointing to an export mechanism in plants similar to Rev-NES-mediated RNP export in other organisms.

  17. Induced mutations and in vitro culture techniques for improving crop plant resistance to diseases

    International Nuclear Information System (INIS)

    1993-12-01

    This co-ordinated research program was undertaken in search of in vitro techniques to increase the resistance of plants to disease. The studies performed under the program ranged from the preparation of materials for mass screening to screening of mutagen-treated cells, tissues, organs or plantlets for resistance to viruses, fungi and other pathogens. The characteristics of the resulting mutants were evaluated to determine the relevance of these techniques for plant breeding. The present document contains the papers presented at the final Research Co-ordination Meeting of the program, as well as a summary of the conclusions and recommendations drawn from the work. The nine individual papers have been input separately to the database. Refs, figs and tabs

  18. The study of cultural objects by nuclear and conventional techniques; Estudio de bienes culturales con tecnicas de caracterizacion nucleares y convencionales

    Energy Technology Data Exchange (ETDEWEB)

    Palacios, Tulio A [Comision Nacional de Energia Atomica, General San Martin (Argentina). Centro Atomico Constituyentes

    2000-07-01

    A survey is given of the techniques that are used at the National Atomic Energy Commission of Argentina for the characterization and study of cultural and archaeological specimens. A short history of these activities is also given. (author)

  19. 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.

  20. 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.

  1. PREFACE: International Conference on the Use of X-ray (and related) Techniques in Arts and Cultural Heritage (XTACH 11)

    Science.gov (United States)

    Hamdan, Nasser; El-Khatib, Sami

    2012-07-01

    The International Conference on the Use of X-Ray (and related) Techniques in Arts and Cultural Heritage (XTACH11) was held on 7 and 8 December 2011 at the American University of Sharjah (AUS) in the United Arab Emirates. The conference was organized in collaboration with the International Atomic Energy Agency (IAEA) and the National X-ray Fluorescence Laboratory (NXFL). The conference was inaugurated by Dr Peter Heath, Chancellor of the American University of Shrjah and attended by Mr Kwaku Aning, deputy Director General of the International Atomic Energy and Ambassador Hamad Al-Kaabi, Ambassador of the UAE to the International Atomic Energy university officials, faculty and students. The conference covered a variety of topics including the use of x-ray and micro beam x-ray analysis, synchrotron based techniques, ion beam and neutron based techniques, optical imaging and mass spectroscopy and chromatography techniques as well as best conservation practices. XTACH11 provided an excellent forum for scientists in the region to interact, exchange ideas and to initiate collaborations with each other as well as with the international community. It showcased some of the latest technical developments in the field of non-destructive testing for the diagnosis and conservation of cultural heritage materials. In addition to the presentations by the invited speakers (Rene van Grieken and K Janssens, University of Antwerp, Belgium; Thomas Calligaro, Centre de Recherche et de Restauration des Musees de France; Stefano Ridolfi, Ars Mensurae, Rome, Italy, and Andrzej Markowicz, IAEA, Austria), a total of 25 other research papers were also presented and discussed. Scientists from many countries participated in the conference: Austria, Belgium, Egypt, Italy, India, Iran, Iraq, Jordan, Lebanon, Qatar, Saudi Arabia, Spain, Sri Lanka, Syria, Thailand, the United Arab Emirates and Yemen. The conference concluded with a Discussion Panel. Thomas Calligaro (Centre de Recherché et de

  2. Multi-technique characterization of gold electroplating on silver substrates for cultural heritage applications

    Science.gov (United States)

    Ortega-Feliu, I.; Ager, F. J.; Roldán, C.; Ferretti, M.; Juanes, D.; Scrivano, S.; Respaldiza, M. A.; Ferrazza, L.; Traver, I.; Grilli, M. L.

    2017-09-01

    This work presents a detailed study of a series of silver plates gilded via electroplating techniques in which the characteristics of the coating gold layers are investigated as a function of the electroplating variables (voltage, time, anode surface and temperature). Some reference samples were coated by radio frequency sputtering in order to compare gold layer homogeneity and effective density. Surface analysis was performed by means of atomic and nuclear techniques (SEM-EDX, EDXRF, PIXE and RBS) to obtain information about thickness, homogeneity, effective density, profile concentration of the gold layers and Au-Ag diffusion profiles. The gold layer thickness obtained by PIXE and EDXRF is consistent with the thickness obtained by means of RBS depth profiling. Electroplated gold mass thickness increases with electroplating time, anode area and voltage. However, electrodeposited samples present rough interfaces and gold layer effective densities lower than the nominal density of Au (19.3 g/cm3), whereas sputtering produces uniform layers with nominal density. These analyses provide valuable information to historians and curators and can help the restoration process of gold-plated silver objects.

  3. Salt Tolerant Mutants in Potato sp. Cara Induced by Gamma Irradiation and Tissue Culture Technique

    International Nuclear Information System (INIS)

    Sharabash, M.T.; Mohamed, A.A.; Ahmed, F.A.; Afifi, A.M.

    2003-01-01

    Sprouts of 2-3 cm length were cut off from potatoes cv. Cara tubers and sterilized. Apical meristemic tips were sown to produce virus free plantlets. After 6-8 weeks, the new plantlets became at 5-7cm heights. Micro-propagation was begun to obtain sufficient number of plantlets. Plantlets obtained from micro-propagation stage were divided into 3 groups to be exposed to 0.0,20 or 40Gy gamma rays. The dose rate was 27.7rad/sec. Irradiated and unirradiated plantlets were trimmed to be with 5-6 nodes and without leaves. They were transplanted onto 1/2MS [1] liquid medium supplemented with sodium chloride at different concentrations, i.e.,0.0, 2000 or 4000 ppm. Healthy plantlets were chosen, and all unfit ones were excluded. The plantlets were sub-cultured six times on the same fresh medium with the same concentrations of NaCI, with insisting to exclude all unfit plantlets. All maintained healthy plantlets, in the sixth vegetative generation (M 1 V 6 ), grown under salinity stress, whatever the concentration of NaCI, were considered as mutant lines

  4. ISOLATION OF MESOPHYLL PROTOPLASTS FROM MEDITERRANEAN WOODY PLANTS FOR THE STUDY OF DNA INTEGRITY UNDER ABIOTIC STRESS

    Directory of Open Access Journals (Sweden)

    Elena Kuzminsky

    2016-08-01

    Full Text Available Abiotic stresses have considerable negative impact on Mediterranean plant ecosystems and better comprehension of the genetic control of response and adaptation of trees to global changes is urgently needed. The Single Cell Gel Electrophoresis assay could be considered a good estimator of DNA damage in an individual eukaryotic cell. This method has been mainly employed in animal tissues, because the plant cell wall represents an obstacle for the extraction of nuclei; moreover, in Mediterranean woody species, especially in the sclerophyll plants, this procedure can be quite difficult because of the presence of sclerenchyma and hardened cells. On the other hand, these plants represent an interesting material to be studied because of the ability of these plants to tolerate abiotic stress. For instance, holm oak (Quercus ilex L. has been selected as the model plant to identify critical levels of O3 for Southern European forests. Consequently, a quantitative method for the evaluation of cell injury of leaf tissues of this species is required. Optimal conditions for high-yield nuclei isolation were obtained by using protoplast technology and a detailed description of the method is provided and discussed. White poplar (Populus alba L. was used as an internal control for protoplast isolation. Such a method has not been previously reported in newly fully developed leaves of holm oak. This method combined with Single Cell Gel Electrophoresis assay represents a new tool for testing the DNA integrity of leaf tissues in higher plants under stress conditions.

  5. Novel thermostable clostridial strains through protoplast fusion for enhanced biobutanol production at higher temperature—preliminary study

    Directory of Open Access Journals (Sweden)

    Muhammad Ferhan

    2016-01-01

    Full Text Available The objective of this study is to improve the thermal stability of clostridium strains for enhanced biobutanol production. Thermostable clostridia species were developed through protoplast fusion between mesophilic clostridial species (i.e., Clostridium beijerinckii and Clostridium acetobutylicum and thermophilic clostridial species (i.e., Clostridium thermocellum. Production of biobutanol was examined in the present preliminary study using the clostridium strains and their protoplast fusants using sugar mixture with composition identical to that of wheat straw acid hydrolysate. Maximum biobutanol production of 9.4 g/L was achieved by a fused strain at 45 °C with total sugar consumption of 66% compared to that at 35 °C (i.e., 8.4 g/L production and 64% total sugar consumption. Glucose and xylose uptake rates were generally higher compared to all other individual sugars in the feedstock. In general, average cell concentrations were in close proximity for all parenting and fused strains at 35 °C; i.e., in the range of 5.12 × 107 to 5.49 × 107 cells/mL. Average cell concentration of fusants between the mesophilic clostridial species and the thermophilic clostridial species slightly increased to ~ 5.62 × 107 cells/mL at a higher temperature of 45 °C. These results, in addition to the ones obtained for the butanol production, demonstrate enhanced thermal stability of both fusants at a higher temperature (45 °C.

  6. Tools, techniques, organisation and culture of the CADD group at Sygnature Discovery.

    Science.gov (United States)

    St-Gallay, Steve A; Sambrook-Smith, Colin P

    2017-03-01

    Computer-aided drug design encompasses a wide variety of tools and techniques, and can be implemented with a range of organisational structures and focus in different organisations. Here we outline the computational chemistry skills within Sygnature Discovery, along with the software and hardware at our disposal, and briefly discuss the methods that are not employed and why. The goal of the group is to provide support for design and analysis in order to improve the quality of compounds synthesised and reduce the timelines of drug discovery projects, and we reveal how this is achieved at Sygnature. Impact on medicinal chemistry is vital to demonstrating the value of computational chemistry, and we discuss the approaches taken to influence the list of compounds for synthesis, and how we recognise success. Finally we touch on some of the areas being developed within the team in order to provide further value to the projects and clients.

  7. [A comparative study of blood culture conventional method vs. a modified lysis/centrifugation technique for the diagnosis of fungemias].

    Science.gov (United States)

    Santiago, Axel Rodolfo; Hernández, Betsy; Rodríguez, Marina; Romero, Hilda

    2004-12-01

    The purpose of this work was to compare the efficacy of blood culture conventional method vs. a modified lysis/centrifugation technique. Out of 450 blood specimens received in one year, 100 where chosen for this comparative study: 60 from patients with AIDS, 15 from leukemic patients, ten from febrile neutropenic patients, five from patients with respiratory infections, five from diabetics and five from septicemic patients. The specimens were processed, simultaneously, according to the above mentioned methodologies with daily inspections searching for fungal growth in order to obtain the final identification of the causative agent. The number (40) of isolates recovered was the same using both methods, which included; 18 Candida albicans (45%), ten Candida spp. (25%), ten Histoplasma capsulatum (25%), and two Cryptococcus neoformans (5%). When the fungal growth time was compared by both methods, growth was more rapid when using the modified lysis/centrifugation technique than when using the conventional method. Statistical analysis revealed a significant difference (pcentrifugation technique showed to be more efficacious than the conventional one, and therefore the implementation of this methodology is highly recommended for the isolation of fungi from blood.

  8. A high efficiency technique for the generation of transgenic sugar beets from stomatal guard cells

    NARCIS (Netherlands)

    Hall, R.D.; Riksen-Bruinsma, T.; Weyens, G.; Rosquin, I.J.; Denys, R.N.; Evans, I.J.; Lathouwers, J.E.; LefObvre, M.P.; Dunwell, J.M.; Tunen, van A.; Krens, F.A.

    1996-01-01

    An optimized protocol has been developed for the efficient and rapid genetic modification of sugar beet (Beta vulgaris L). A polyethylene glycol- mediated DNA transformation technique could be applied to protoplast populations enriched specifically for a single totipotent cell type derived from

  9. Efforts towards domestication of wild tomato (solanum pimpinellifolium L.) using mutation and in vitro culture techniques

    International Nuclear Information System (INIS)

    Quartey, E. K.

    2010-06-01

    antioxidant properties (90% DPPH inhibition at 75 and 100 ml volumes) compared to the control (84.4% DPPH inhibition at 100 ml). Fruits of variant line C-2*1(4) (yellow fruits) recorded higher total solids of 17.9% and the lowest pH value of 4.17 compared to 12.8% and 4.36 for fruits from un-irradiated plants. The highest amounts of total soluble solids (7%) were contained in fruits harvested from variant lines G-1 *4(4) (light red fruits) and P-2*3(29) (deep red fruits) as against 5.6% for fruits from control plants. Similarly, fruits of line X- 1*4(9) (deep red fruits) contained highest amounts of potassium (7.31 g/kg in pericarp; 4.80 g/kg in pulp + seeds), sodium (106.3 mg/kg in pericarp; 120.6 mg/kg in pulp + seeds) and manganese (10.03 mg/kg in pericarp; 39.21 mg/kg in pulp + seeds). Fruits of lines G-1*4(4), SA-2*1(27) (deep red fruits) and C-2*1(4) contained highest amounts of magnesium (0.17 g/kg in pericarp; 0.34 g/kg in pulp + seeds), calcium (0.22 g/kg in pericarp and pulp + seeds) and vanadium (0.28 mg/kg in pericarp; 0.35 mg/kg in pulp + seeds) respectively. Soft vines propagated in vivo recorded optimum performance on a substrate of 3:1:1 of topsoil, cow dung and coconut fibre respectively (91.7% shoot regeneration). Hormone-free MS basal medium supported full plantlet regeneration (100%) from nodal cuttings under in vitro culture conditions. Hypocotyls proved to be better explants compared to cotyledons for callus induction using same MS medium supplemented with phytohormones at concentrations of 1.0 mg/L BAP + 1.5 mg/L 2, 4-D under 16/8-hr light/dark after three (3) weeks in culture. Significant differences (p≤0.05) were recorded for most of the traits studied among the selected variants. These include lycopene content, total solids and total soluble solids. There were also improvements in the macro and micro elemental composition of fruits of the selected variants. Additionally lower pH value, which is a more acceptable characteristic for processing of

  10. Physicochemical characterisation of pottery from the Vinča culture, Serbia, regarding the firing temperature and decoration techniques

    Directory of Open Access Journals (Sweden)

    Perišić Nebojša

    2016-01-01

    Full Text Available A study of decorated Neolithic pottery samples from excavation site Pločnik, Serbia, was performed using X-ray powder diffraction (XRPD, Fourier transform infrared (FTIR and X-ray fluorescence (XRF spectroscopy. Investigated samples belong to the era of the Vinča culture which existed at the central Balkan region from mid VI until the first half of V millennium BCE. The mineralogical composition of pottery samples and comparison of investigated pottery with thermally treated local clay indicated firing temperature in the range from 600 to 800°C. Two different types of white pigments have been identified in white incrusted decorations: calcium carbonate and Bone White (composed of crushed bones. This is the first evidence of use of bones for decorations in Vinča culture pottery from excavation site Pločnik. In addition to this, it was revealed that the potters used the iron reduction technique for obtaining the black decorations. [Projekat Ministartsva nauke Republike Srbije, br. 177021 I br. 177012

  11. Creating a classroom culture that supports the common core teaching questioning, conversation techniques, and other essential skills

    CERN Document Server

    Harris, Bryan

    2014-01-01

    Is your classroom culture conducive to the expectations of the Common Core? Teaching content is not enough; students need a classroom structure and atmosphere that will help them learn key academic skills. This practical book will show you how to transform your classroom culture, raise the level of rigor, encourage higher-level questioning and critical thinking, and promote academic discussions. You will also find out how to adjust your classroom management techniques so that students learn to regulate themselves while completing these higher-level tasks. Special Features in Each Chapter: Key Idea-a summary of the essential idea that will be addressed in the chapter Practical strategies-a variety of easy-to-implement ideas that you can try right away Connections to the Common Core State Standards-how the skills taught in this book will help students meet the standards Reflection Questions-thoughtful questions that will help teachers apply their learning to their own classrooms. These questions can be answered...

  12. Integration of Geomatics Techniques for Digitizing Highly Relevant Geological and Cultural Heritage Sites: the Case of San Leo (italy)

    Science.gov (United States)

    Girelli, V. A.; Borgatti, L.; Dellapasqua, M.; Mandanici, E.; Spreafico, M. C.; Tini, M. A.; Bitelli, G.

    2017-08-01

    The research activities described in this contribution were carried out at San Leo (Italy). The town is located on the top of a quadrangular rock slab affected by a complex system of fractures and has a wealth of cultural heritage, as evidenced by the UNESCO's nomination. The management of this fragile set requires a comprehensive system of geometrical information to analyse and preserve all the geological and cultural features. In this perspective, the latest Geomatics techniques were used to perform some detailed surveys and to manage the great amount of acquired geometrical knowledge of both natural (the cliff) and historical heritage. All the data were also georeferenced in a unique reference system. In particular, high accurate terrestrial laser scanner surveys were performed for the whole cliff, in order to obtain a dense point cloud useful for a large number of geological studies, among others the analyses of the last rockslide by comparing pre- and post-event data. Moreover, the geometrical representation of the historical centre was performed using different approaches, in order to generate an accurate DTM and DSM of the site. For these purposes, a large scale numerical map was used, integrating the data with GNSS and laser surveys of the area. Finally, many surveys were performed with different approaches on some of the most relevant monuments of the town. In fact, these surveys were performed by terrestrial laser scanner, light structured scanner and photogrammetry, the last mainly applied with the Structure from Motion approach.

  13. Improved enrichment culture technique for methane-oxidizing bacteria from marine ecosystems: the effect of adhesion material and gas composition.

    Science.gov (United States)

    Vekeman, Bram; Dumolin, Charles; De Vos, Paul; Heylen, Kim

    2017-02-01

    Cultivation of microbial representatives of specific functional guilds from environmental samples depends largely on the suitability of the applied growth conditions. Especially the cultivation of marine methanotrophs has received little attention, resulting in only a limited number of ex situ cultures available. In this study we investigated the effect of adhesion material and headspace composition on the methane oxidation activity in methanotrophic enrichments obtained from marine sediment. Addition of sterilized natural sediment or alternatively the addition of acid-washed silicon dioxide significantly increased methane oxidation. This positive effect was attributed to bacterial adhesion on the particles via extracellular compounds, with a minimum amount of particles required for effect. As a result, the particles were immobilized, thus creating a stratified environment in which a limited diffusive gas gradients could build up and various microniches were formed. Such diffusive gas gradient might necessitate high headspace concentrations of CH 4 and CO 2 for sufficient concentrations to reach the methane-oxidizing bacteria in the enrichment culture technique. Therefore, high concentrations of methane and carbon dioxide, in addition to the addition of adhesion material, were tested and indeed further stimulated methane oxidation. Use of adhesion material in combination with high concentrations of methane and carbon dioxide might thus facilitate the cultivation and subsequent enrichment of environmentally important members of this functional guild. The exact mechanism of the observed positive effects on methane oxidation and the differential effect on methanotrophic diversity still needs to be explored.

  14. Biosynthesis of acid phosphatase of baker's yeast . Characterization of a protoplast-bound fraction containing precursors of the exo-enzyme

    NARCIS (Netherlands)

    Boer, Pieter; Rijn, Herman J.M. van; Reinking, A.; Steyn-Parvé, Elizabeth P.

    1975-01-01

    1. 1.|Yest protoplasts, secreting acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum) EC 3.1.3.2) contain a small amount of firmly bound enzyme, even after lysis (Van Rijn, H.J.M.; Boer, P. and Steyn-Parvé, E.P. (1972) Biochim. Biophys. Acta 268, 431–441). The major part

  15. Studies on protein synthesis by protoplasts of saccharomyces carlsbergensis III. Studies on the specificity and the mechanism of the action of ribonuclease on protein synthesis

    NARCIS (Netherlands)

    Kloet, S.R. de; Dam, G.J.W. van; Koningsberger, V.V.

    1962-01-01

    In this paper, the experimental results are presented of a continued study on the specificity and the mechanism of the inhibition by ribonuclease of protein synthesis in protoplasts of Saccharomyces carlsbergensis. By comparing the effects of native pancreatic ribonuclease with those of

  16. Naturally Induced Secretions of the Potato Cyst Nematode Co-stimulate the Proliferation of Both Tobacco Leaf Protoplasts and Human Peripheral Blood Mononuclear Cells

    NARCIS (Netherlands)

    Goverse, A.; Rouppe van der Voort, J.N.A.M.; Rouppe van der voort, C.; Kavelaars, A.; Smant, G.; Schots, A.; Bakker, J.; Helder, J.

    1999-01-01

    Naturally induced secretions from infective juveniles of the potato cyst nematode Globodera rostochiensis co-stimulate the proliferation of tobacco leaf protoplasts in the presence of the synthetic phytohormones α-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP). With the use of a

  17. Studies on protein synthesis by protoplasts of Saccharomyces carlsbergensis II. Reversal of the RNase effect of protein synthesis by polymethacrylic acid

    NARCIS (Netherlands)

    Kloet, S.R. de; Wermeskerken, R.K.A. van; Koningsberger, V.V.

    1961-01-01

    The ribonuclease inhibited protein synthesis and respiration of yeast protoplasts can be restored by the addition of several polyanionic compounds, among which polymethacrylic acid proved to be the most effective one. The results of preliminary experiments with the ultracentrifuge indicate a

  18. Spatial and temporal single-cell volume estimation by a fluorescence imaging technique with application to astrocytes in primary culture

    Science.gov (United States)

    Khatibi, Siamak; Allansson, Louise; Gustavsson, Tomas; Blomstrand, Fredrik; Hansson, Elisabeth; Olsson, Torsten

    1999-05-01

    Cell volume changes are often associated with important physiological and pathological processes in the cell. These changes may be the means by which the cell interacts with its surrounding. Astroglial cells change their volume and shape under several circumstances that affect the central nervous system. Following an incidence of brain damage, such as a stroke or a traumatic brain injury, one of the first events seen is swelling of the astroglial cells. In order to study this and other similar phenomena, it is desirable to develop technical instrumentation and analysis methods capable of detecting and characterizing dynamic cell shape changes in a quantitative and robust way. We have developed a technique to monitor and to quantify the spatial and temporal volume changes in a single cell in primary culture. The technique is based on two- and three-dimensional fluorescence imaging. The temporal information is obtained from a sequence of microscope images, which are analyzed in real time. The spatial data is collected in a sequence of images from the microscope, which is automatically focused up and down through the specimen. The analysis of spatial data is performed off-line and consists of photobleaching compensation, focus restoration, filtering, segmentation and spatial volume estimation.

  19. Expression of human immunodeficiency virus (HIV) in naturally infected peripheral blood mononuclear cells: comparison of a standard co-culture technique with a newly developed microculture method.

    Science.gov (United States)

    Eberlein, B; Baur, A; Neundorfer, M; Jahn, G

    1991-05-01

    Peripheral blood mononuclear cells (PBMCs) from 29 patients infected with human immunodeficiency virus (HIV) were cultured by two different methods. One was the standard co-culture technique, the other a newly developed microculture method. In this assay 10(6) PBMCs were cultivated in 250 microliters medium, no activating agents or allogeneic cells were present. P24 antigen production measured by this method was found in 7 out of 11 PBMC cultures of patients in the Walter Reed (WR) stage 1 or 2, whereas only 4 samples were positive by the co-culture procedure. Cultures from patients in the later stages of the disease (WR 5/6) showed a higher p24 production by the co-culture method than by the microculture assay. It is assumed that rapidly growing HIV strains can be better assessed by the co-culture method which may select for these strains. P24 expression can be more easily obtained by the microculture technique even in cases where slowly replicating strains may be present. In conclusion, results from the microculture procedure described may be a useful supplementation to findings observed by the co-culture method.

  20. Differential compartmentation of sucrose and gentianose in the cytosol and vacuoles of storage root protoplasts from Gentiana Lutea L.

    Science.gov (United States)

    Keller, F; Wiemken, A

    1982-12-01

    The storage roots of perennial Gentiana lutea L.plants contain several sugars. The predominant carbohydrate reserve is gentianose (β-D-glucopyranosyl-(1 → 6)-α-D-glucopyranosyl-(1 ↔ 2)-β-D-fructofuranoside). Vacuoles were isolated from root protoplasts and purified through a betaine density gradient. The yield was about 75%. Gentianose and gentiobiose were localized to 100% in the vacuoles, fructose and glucose to about 80%, and sucrose to only about 50%. Taking the volumes of the vacuolar and extravacuolar (cytosolic) compartments into account it is inferred that gentianose is located exclusively in the vacuoles, whilst sucrose is much more concentrated in the cytosol where it may play a role as a cryoprotectant. The concentration of fructose and glucose appeared to be similar on both sides of the tonoplast.

  1. Construction and evaluation of an exopolysaccharide-producing engineered bacterial strain by protoplast fusion for microbial enhanced oil recovery.

    Science.gov (United States)

    Sun, Shanshan; Luo, Yijing; Cao, Siyuan; Li, Wenhong; Zhang, Zhongzhi; Jiang, Lingxi; Dong, Hanping; Yu, Li; Wu, Wei-Min

    2013-09-01

    Enterobacter cloacae strain JD, which produces water-insoluble biopolymers at optimal temperature of 30°C, and a thermophilic Geobacillus strain were used to construct an engineered strain for exopolysaccharide production at high temperatures by protoplast fusion. The obtained fusant strain ZR3 produced exopolysaccharides at up to 45°C with optimal growth temperature at 35°C. The fusant produced exopolysaccharides of approximately 7.5 g/L or more at pH between 7.0 and 9.0. The feasibility of the enhancement of crude oil recovery with the fusant was tested in a sand-packed column at 40°C. The results demonstrated that bioaugmentation of the fusant was promising approach for MEOR. Mass growth of the fusant was confirmed in fermentor tests. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. Phytochrome-mediated responses of cells and protoplasts of green calli obtained from the leaves of a CAM plant.

    Science.gov (United States)

    Mricha, A; Brulfert, J; Pierre, J N; Queiroz, O

    1990-04-01

    Green callus obtained from leaves of the CAM-inducible plant Kalanchoe blossfeldiana cv. Montezuma has previously been shown to perform C3-type photosynthesis under 16-h days and to shift to crassulacean acid metabolism (CAM) under 9-h days. The utilization of photoperiodic regimes (i.e. night interruptions by 30 min red light) established that CAM induction in the callus was under the control of phytochrome, as shown by measurements of CAM criteria: phosphoenolpyruvate carboxylase activity and malic acid pools. Short-term responsiveness of the callus cells to phytochrome modulations by monochromatic radiations was also established by the rapid changes observed in the diameter of the callus-derived protoplasts. These results provide further evidence that whole plant correlations are not necessary for phytochrome operativity.

  3. Growth of Avena Coleoptiles and pH Drop of Protoplast Suspensions Induced by Chlorinated Indoleacetic Acids

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen; Doll, Hans; Böttger, M.

    1978-01-01

    -auxins. Some of the derivatives were compared for their effect on pH decline in stem protoplast suspensions of Helianthus annuus L. and Pisum sativum L. The change of pH occurs without a lag period or with only a very short one. Derivatives which are very active in the Avena straight growth assay cause......Several indoleacetic acids, substituted in the benzene ring, were compared in the Avena straight growth bioassay. 4-Chloroindoleacetic acid, a naturally occurring plant hormone, is one of the strongest hormones in this bioassay. With an optimum at 10-6 mol l-1, it is more active than indoleacetic...... a larger pH decline than indoleacetic acid, while inactive derivatives cause effectively no pH decline....

  4. Comparison Between Laser Scanning and Automated 3d Modelling Techniques to Reconstruct Complex and Extensive Cultural Heritage Areas

    Science.gov (United States)

    Fassi, F.; Fregonese, L.; Ackermann, S.; De Troia, V.

    2013-02-01

    In Cultural Heritage field, the necessity to survey objects in a fast manner, with the ability to repeat the measurements several times for deformation or degradation monitoring purposes, is increasing. In this paper, two significant cases, an architectonical one and an archaeological one, are presented. Due to different reasons and emergency situations, the finding of the optimal solution to enable quick and well-timed survey for a complete digital reconstruction of the object is required. In both cases, two survey methods have been tested and used: a laser scanning approach that allows to obtain high-resolution and complete scans within a short time and a photogrammetric one that allows the three-dimensional reconstruction of the object from images. In the last months, several methodologies, including free or low cost techniques, have arisen. These kinds of software allow the fully automatically three-dimensional reconstruction of objects from images, giving back a dense point cloud and, in some case, a surfaced mesh model. In this paper some comparisons between the two methodologies above mentioned are presented, using the example of some real cases of study. The surveys have been performed by employing both photogrammetry and laser scanner techniques. The methodological operational choices, depending on the required goal, the difficulties encountered during the survey with these methods, the execution time (that is the key parameter), and finally the obtained results, are fully described and examinated. On the final 3D model, an analytical comparison has been made, to analyse the differences, the tolerances, the possibility of accuracy improvement and the future developments.

  5. Evaluation of different selective media and culturing techniques for the quantification of Campylobacter ssp. from broiler litter.

    Science.gov (United States)

    Kiess, A S; Parker, H M; McDaniel, C D

    2010-08-01

    Poultry is a major reservoir for Campylobacter, the leading cause of foodborne illness in the United States, but how broilers become initially colonized is still under debate. Broiler litter is a potential source, but the best technique for quantifying Campylobacter from litter is still unknown. Therefore, our objectives were to determine if certain media are more selective for quantifying Campylobacter and if enrichment allows for the detection of stressed or viable but nonculturable cells from broiler litter samples. In this trial, 5 media and 2 culturing techniques were used to enumerate Campylobacter from broiler litter. The media used were campy-Line agar (CLA), campy-cefex agar (CCA), modified CCA, Campylobacter agar plates (CAP), and modified charcoal cefoperazone deoxycholate agar. Litter samples were obtained from a commercial broiler house. Each sample was equally divided and diluted 10-fold into peptone, for direct plating, or 4-fold into Campylobacter enrichment broth. Samples diluted in peptone were direct-plated onto each media and incubated under microaerophilic conditions for 48 h at 42 degrees C. Samples diluted in enrichment broth were incubated under the same conditions for 24 h, then further diluted to 10-fold before plating. Plates from enriched samples were incubated for an additional 24 h after plating. After incubation, all plates (direct and enriched) were counted and presumptive positive colonies were confirmed using a Campylobacter latex agglutination kit. Results indicated that there was no difference in the ability of any of the selective media tested to grow Campylobacter. Direct-plated samples had a higher Campylobacter isolation rate compared with enriched samples. The CLA and CAP were able to suppress total bacterial growth better than modified charcoal cefoperazone deoxycholate, modified CCA, and CCA. The CLA and CAP were the only media able to detect total bacterial population shifts over time. In conclusion, it is important

  6. Translation Techniques

    OpenAIRE

    Marcia Pinheiro

    2015-01-01

    In this paper, we discuss three translation techniques: literal, cultural, and artistic. Literal translation is a well-known technique, which means that it is quite easy to find sources on the topic. Cultural and artistic translation may be new terms. Whilst cultural translation focuses on matching contexts, artistic translation focuses on matching reactions. Because literal translation matches only words, it is not hard to find situations in which we should not use this technique.  Because a...

  7. [Prokaryotic community of subglacial bottom sediments of Antarctic Lake Untersee: detection by cultural and direct microscopic techniques].

    Science.gov (United States)

    Muliukin, A L; Demkina, E V; Manucharova, N A; Akimov, V N; Andersen, D; McKay, C; Gal'chenko, V F

    2014-01-01

    The heterotrophic mesophilic component was studied in microbial communities of the samples of frozen regolith collected from the glacier near Lake Untersee collected in 2011 during the joint Russian-American expedition to central Dronning Maud Land (Eastern Antarctica). Cultural techniques revealed high bacterial numbers in the samples. For enumeration of viable cells, the most probable numbers (MPN) method proved more efficient than plating on agar media. Fluorescent in situ hybridization with the relevant oligonucleotide probes revealed members of the groups Eubacteria (Actinobacteria, Firmicutes) and Archaea. Application of the methods of cell resuscitation, such as the use of diluted media and prevention of oxidative stress, did not result in a significant increase in the numbers of viable cells retrieved form subglacial sediment samples. Our previous investigations demonstrated the necessity for special procedures for efficient reactivation of the cells from microbial communities of preserved fossil soil and permafrost samples collected in the Arctic zone. The differences in response to the special resuscitation procedures may reflect the differences in the physiological and morphological state of bacterial cells in microbial communities subject to continuous or periodic low temperatures and dehydration.

  8. Results of Applying Cultural Domain Analysis Techniques and Implications for the Design of Complementary Feeding Interventions in Northern Senegal.

    Science.gov (United States)

    Zobrist, Stephanie; Kalra, Nikhila; Pelto, Gretel; Wittenbrink, Brittney; Milani, Peiman; Diallo, Abdoulaye Moussa; Ndoye, Tidiane; Wone, Issa; Parker, Megan

    2017-12-01

    Designing effective nutrition interventions for infants and young children requires knowledge about the population to which the intervention is directed, including insights into the cognitive systems and values that inform caregiver feeding practices. To apply cultural domain analysis techniques in the context of implementation research for the purpose of understanding caregivers' knowledge frameworks in Northern Senegal with respect to infant and young child (IYC) feeding. This study was intended to inform decisions for interventions to improve infant and young child nutrition. Modules from the Focused Ethnographic Study for Infant and Young Child Feeding Manual were employed in interviews with a sample of 126 key informants and caregivers from rural and peri-urban sites in the Saint-Louis region of northern Senegal. Descriptive statistics, cluster analysis, and qualitative thematic analysis were used to analyze the data. Cluster analysis showed that caregivers identified 6 food clusters: heavy foods, light foods, snack foods, foraged foods, packaged foods, and foods that are good for the body. The study also revealed similarities and differences between the 2 study sites in caregivers' knowledge frameworks. The demonstration of differences between biomedical concepts of nutrition and the knowledge frameworks of northern Senegalese women with regard to IYC feeding highlights the value of knowledge about emic perspectives of local communities to help guide decisions about interventions to improve nutrition.

  9. Degradation measurement of bronzes, a recent application of the TLA techniques in the field of cultural heritage

    International Nuclear Information System (INIS)

    Danesi, P.R.; Laguzzi, G.; Luvidi, L

    2001-01-01

    The thin layer technique is normally applied in wear and corrosion monitoring of mechanical and plant components. A recent applications refers to the field of Cultural Heritage. In particular thought national and international research programmes TLA is used as a sensitivity toll in determining the properties towards corrosion of new bronze alloys to be used for artistic purposes, as in manufacturing of outdoor artworks. One activated and exposed to natural or artificial corrosive environments the bronze surface is treated with pick ling solutions to remove the corrosion products, The corresponding thickness loss is determined by the relevant γ-activity loss. The method sensitivity allows of the thickness loss evaluation in the order micrometers of lower. Besides comparative analyses of the behaviour of different bronze alloys, this method is also applied in determining the effectiveness of corrosion inhibitor or protective films, normally used on outdoor bronze sculptures. The metal activation is carried out by using a cyclotron accelerated proton beam (11.5 MeV) which products on the bronze surface and along a defined depth γemitting radionuclide 65 Zn (t 1/2 = 244 days). In the present paper, bronze corrosion results obtained in different environmental conditions as outdoor exposures and artificial weathering experiments are reported

  10. Comparison of different sampling techniques and of different culture methods for detection of group B streptococcus carriage in pregnant women

    Directory of Open Access Journals (Sweden)

    Verhelst Rita

    2010-09-01

    Full Text Available Abstract Background Streptococcus agalactiae (group B streptococcus; GBS is a significant cause of perinatal and neonatal infections worldwide. To detect GBS colonization in pregnant women, the CDC recommends isolation of the bacterium from vaginal and anorectal swab samples by growth in a selective enrichment medium, such as Lim broth (Todd-Hewitt broth supplemented with selective antibiotics, followed by subculture on sheep blood agar. However, this procedure may require 48 h to complete. We compared different sampling and culture techniques for the detection of GBS. Methods A total of 300 swabs was taken from 100 pregnant women at 35-37 weeks of gestation. For each subject, one rectovaginal, one vaginal and one rectal ESwab were collected. Plating onto Columbia CNA agar (CNA, group B streptococcus differential agar (GBSDA (Granada Medium and chromID Strepto B agar (CA, with and without Lim broth enrichment, were compared. The isolates were confirmed as S. agalactiae using the CAMP test on blood agar and by molecular identification with tDNA-PCR or by 16S rRNA gene sequence determination. Results The overall GBS colonization rate was 22%. GBS positivity for rectovaginal sampling (100% was significantly higher than detection on the basis of vaginal sampling (50%, but not significantly higher than for rectal sampling (82%. Direct plating of the rectovaginal swab on CNA, GBSDA and CA resulted in detection of 59, 91 and 95% of the carriers, respectively, whereas subculturing of Lim broth yielded 77, 95 and 100% positivity, respectively. Lim broth enrichment enabled the detection of only one additional GBS positive subject. There was no significant difference between GBSDA and CA, whereas both were more sensitive than CNA. Direct culture onto GBSDA or CA (91 and 95% detected more carriers than Lim broth enrichment and subculture onto CNA (77%. One false negative isolate was observed on GBSDA, and three false positives on CA. Conclusions In

  11. Component Analysis and Identification of Black Tahitian Cultured Pearls From the Oyster Pinctada margaritifera Using Spectroscopic Techniques

    Science.gov (United States)

    Shi, L.; Wang, Y.; Liu, X.; Mao, J.

    2018-03-01

    Raman spectroscopy, ultraviolet, visible, and near infrared (UV-Vis-NIR) reflectance spectroscopy, and X-ray fluorescence (XRF) spectroscopy were used to characterize black Tahitian cultured pearls and imitations of these saltwater cultured pearls produced by γ-irradiation, and by coloring of cultured pearls with silver nitrate or organic dyes. Raman spectra indicated that aragonite was the major constituent of these four types of pearl. Using Raman spectroscopy at an excitation wavelength of 514 nm, black Tahitian cultured pearls exhibited characteristic 1100-1700 cm-1 bands. These bands were attributed to various organic components, including conchiolin and other black biological pigments. The peaks shown by saltwater cultured pearls colored with organic dyes varied with the type of dye used. Tahitian cultured and organic-dye-treated saltwater cultured pearls were easily identified by Raman spectroscopy. UV-Vis-NIR reflectance spectra showed bands at 408, 497, and 700 nm derived from porphyrin pigment and other black pigments. The spectra of dye-treated black saltwater pearls showed absorption peaks at 216, 261, 300, and 578 nm. The 261-nm absorption band disappeared from the spectra of γ-irradiated saltwater cultured pearls. This suggests the degradation of conchiolin in the γ-irradiated saltwater cultured pearls. XRF analysis revealed the presence of Ag on the surface of silver nitrate-dyed saltwater cultured pearls.

  12. Screening for sweet potato (Ipomoea batatas (L.)) viruses and their elimination using thermotheraphy-meristem tip culture technique

    International Nuclear Information System (INIS)

    Arkorful, E.

    2012-11-01

    Despite its high potential for food security, production of sweet potato is constrained by viruses which reduce yield by 90%. It is therefore essential to screen for, identify and eliminate these viruses in elite clones before dissemination to farmers. In this study, visual symptomatology and PCR-based techniques were used to identify sweet potato viruses. Visual symptomatology revealed virus associated symptoms ranging from vein clearing, interveinal chlorosis, chlorotic spots, upward curling on leaf edges, leaf narrowing and distortion, purpling, blistering, reduction of the leaf blades and general leaf yellowing in all 22 accessions grown on the field. Disease Incidence (DI) significantly (p≤0.05) varied between accessions with US003 having the lowest (20%) while ten accessions had 90% DI at the end of the study. Index of symptom severity of all plants (ISSap) ranged from 1.08±0.09 to 3.67 ±0.11 with VOTCR003 having the lowest suggesting that it is a moderately susceptible accession while VOTCR002 had the highest suggesting that it is susceptible to viral diseases. Contrarily, index of symptom severity of diseased plants (ISSdp) ranged from 2.00±0.25 to 3.75±0.32. The accession VOTCR002 had the highest ISSdp. Visual symptomatology showed that VOTCR002 had the highest DI, ISSap and ISSdp suggesting that it is highly susceptible to viral diseases. Ten severely infected accessions were tested for Sweet Potato leaf curl virus (SPLCV), Sweet Potato chlorotic stunt virus (SPCSV), Sweet potato feathery mottle virus (SPFMV) and Sweet Potato mild mottle virus (SPMMV) using PCR and RT-PCR techniques. RT-PCR did not amplify any of the virus genomes due to prolonged storage enzymes, In contrast, PCR detected SPLCV in 30% of the accessions. Plants infected with SPLCV were grown in the chamber at 35 degrees celsius for 4 weeks followed by meristem top culture. The regenerants were indexed after ten weeks for SPLCV. Fifty two percent (52.385 od the regenerants were

  13. Arabidopsis Regenerating Protoplast: A Powerful Model System for Combining the Proteomics of Cell Wall Proteins and the Visualization of Cell Wall Dynamics

    Science.gov (United States)

    Yokoyama, Ryusuke; Kuki, Hiroaki; Kuroha, Takeshi; Nishitani, Kazuhiko

    2016-01-01

    The development of a range of sub-proteomic approaches to the plant cell wall has identified many of the cell wall proteins. However, it remains difficult to elucidate the precise biological role of each protein and the cell wall dynamics driven by their actions. The plant protoplast provides an excellent means not only for characterizing cell wall proteins, but also for visualizing the dynamics of cell wall regeneration, during which cell wall proteins are secreted. It therefore offers a unique opportunity to investigate the de novo construction process of the cell wall. This review deals with sub-proteomic approaches to the plant cell wall through the use of protoplasts, a methodology that will provide the basis for further exploration of cell wall proteins and cell wall dynamics. PMID:28248244

  14. Detection of mycoplasmas in urethral swabs from HIV-1 infected patients and control individuals using culture techniques and polymerase chain reaction

    Directory of Open Access Journals (Sweden)

    CUNHA Regina Ayr Florio da

    1998-01-01

    Full Text Available The objective of the present study was to determine the prevalence of certain mycoplasma species, i.e., Mycoplasma hominis, Ureaplasma urealyticum and Mycoplasma penetrans, in urethral swabs from HIV-1 infected patients compared to swabs from a control group. Mycoplasmas were detected by routine culture techniques and by the Polymerase Chain Reaction (PCR technique, using 16SrRNA generic primers of conserved region and Mycoplasma penetrans specific primers. The positivity rates obtained with the two methods were comparable. Nevertheless, PCR was more sensitive, while the culture techniques allowed the quantification of the isolates. The results showed no significant difference (p < 0.05 in positivity rates between the methods used for mycoplasma detection.

  15. Arabidopsis Regenerating Protoplast: A Powerful Model System for Combining the Proteomics of Cell Wall Proteins and the Visualization of Cell Wall Dynamics

    OpenAIRE

    Yokoyama, Ryusuke; Kuki, Hiroaki; Kuroha, Takeshi; Nishitani, Kazuhiko

    2016-01-01

    The development of a range of sub-proteomic approaches to the plant cell wall has identified many of the cell wall proteins. However, it remains difficult to elucidate the precise biological role of each protein and the cell wall dynamics driven by their actions. The plant protoplast provides an excellent means not only for characterizing cell wall proteins, but also for visualizing the dynamics of cell wall regeneration, during which cell wall proteins are secreted. It therefore offers a uni...

  16. Synthesis of viral DNA forms in Nicotiana plumbaginifolia protoplasts inoculated with cassava latent virus (CLV); evidence for the independent replication of one component of the CLV genome.

    OpenAIRE

    Townsend, R; Watts, J; Stanley, J

    1986-01-01

    Totipotent leaf mesophyll protoplasts of Nicotiana plumbaginifolia, Viviani were inoculated with cassava latent virus (CLV) or with full length copies of CLV genomic DNAs 1 and 2 excised from replicative forms of M13 clones. Virus specific DNAs began to appear 48-72h after inoculation with virus or cloned DNAs, coincident with the onset of host cell division. Infected cells accumulated supercoiled forms of DNAs 1 and 2 as well as progeny single-stranded (ss) virion (+) sense DNAs representing...

  17. Site-directed mutagenesis in Petunia × hybrida protoplast system using direct delivery of purified recombinant Cas9 ribonucleoproteins.

    Science.gov (United States)

    Subburaj, Saminathan; Chung, Sung Jin; Lee, Choongil; Ryu, Seuk-Min; Kim, Duk Hyoung; Kim, Jin-Soo; Bae, Sangsu; Lee, Geung-Joo

    2016-07-01

    Site-directed mutagenesis of nitrate reductase genes using direct delivery of purified Cas9 protein preassembled with guide RNA produces mutations efficiently in Petunia × hybrida protoplast system. The clustered, regularly interspaced, short palindromic repeat (CRISPR)-CRISPR associated endonuclease 9 (CRISPR/Cas9) system has been recently announced as a powerful molecular breeding tool for site-directed mutagenesis in higher plants. Here, we report a site-directed mutagenesis method targeting Petunia nitrate reductase (NR) gene locus. This method could create mutations efficiently using direct delivery of purified Cas9 protein and single guide RNA (sgRNA) into protoplast cells. After transient introduction of RNA-guided endonuclease (RGEN) ribonucleoproteins (RNPs) with different sgRNAs targeting NR genes, mutagenesis at the targeted loci was detected by T7E1 assay and confirmed by targeted deep sequencing. T7E1 assay showed that RGEN RNPs induced site-specific mutations at frequencies ranging from 2.4 to 21 % at four different sites (NR1, 2, 4 and 6) in the PhNR gene locus with average mutation efficiency of 14.9 ± 2.2 %. Targeted deep DNA sequencing revealed mutation rates of 5.3-17.8 % with average mutation rate of 11.5 ± 2 % at the same NR gene target sites in DNA fragments of analyzed protoplast transfectants. Further analysis from targeted deep sequencing showed that the average ratio of deletion to insertion produced collectively by the four NR-RGEN target sites (NR1, 2, 4, and 6) was about 63:37. Our results demonstrated that direct delivery of RGEN RNPs into protoplast cells of Petunia can be exploited as an efficient tool for site-directed mutagenesis of genes or genome editing in plant systems.

  18. (Review paper) The role of biotechnology in crop improvement ...

    African Journals Online (AJOL)

    Pollen tube-pathway, Electroporation, Liposome fusion were considered. While in Bio culture Techniques. Embryo Rescue. Tissue Culture, Anther/Pollen Culture and Protoplast Culture were looked at. The Author also ... HOW TO USE AJOL.

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

    Science.gov (United States)

    Kaneko, Ai; Sankai, Yoshiyuki

    2014-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Ai Kaneko

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

  1. Use of Tissue Culture Techniques for Producing Virus-Free Plant in Garlic and Their Identification through Real-Time PCR

    Directory of Open Access Journals (Sweden)

    Hatıra Taşkın

    2013-01-01

    Full Text Available This study was performed for comparison of meristem culture technique with shoot tip culture technique for obtaining virus-free plant, comparison of micropropagation success of two different nutrient media, and determination of effectiveness of real-time PCR assay for the detection of viruses. Two different garlic species (Allium sativum and Allium tuncelianum and two different nutrient media were used in this experiment. Results showed that Medium 2 was more successful compared to Medium 1 for both A. tuncelianum and A. sativum (Kastamonu garlic clone. In vitro plants obtained via meristem and shoot tip cultures were tested for determination of onion yellow dwarf virus (OYDV and leek yellow stripe virus (LYSV through real-time PCR assay. In garlic plants propagated via meristem culture, we could not detect any virus. OYDV and LYSV viruses were detected in plants obtained via shoot tip culture. OYDV virus was observed in amount of 80% and 73% of tested plants for A. tuncelianum and A. sativum, respectively. LYSV virus was found in amount of 67% of tested plants of A. tuncelianum and in amount of 87% of tested plants of A. sativum in this study.

  2. Effect of specimen collection techniques, transport media, and incubation of cultures on the detection rate of Helicobacter pylori

    NARCIS (Netherlands)

    van der Hulst, R. W.; Verheul, S. B.; Weel, J. F.; Gerrits, Y.; ten Kate, F. J.; Dankert, J.; Tytgat, G. N.

    1996-01-01

    Culture and histologic examination are considered "gold standard" methods for the detection of Helicobacter pylori, but discrepancies may occur with either method. Failure to detect Helicobacter pylori may be due to sampling error, inappropriate transport or culture media, or insufficient duration

  3. Simplified non-cultured non-trypsinised epidermal cell graft technique followed by psoralen and ultraviolet a light therapy for stable vitiligo

    Directory of Open Access Journals (Sweden)

    Dilip Kachhawa

    2017-01-01

    Full Text Available Background and Aims: Stable vitiligo can be treated by various surgical procedures. Non-cultured melanocyte grafting techniques were developed to overcome the time-consuming process of culture while at the same time providing acceptable results. All the techniques using non-cultured melanocyte transfer involve trypsinisation as an integral step. Jodhpur technique used by the author is autologous, non-cultured, non-trypsinised, epidermal cell grafting. Settings and Design: The study was conducted on patients visiting the dermatology outpatient department of a tertiary health centre in Western Rajasthan. Materials and Methods: At the donor site, mupirocin ointment was applied and dermabrasion was done with the help of micromotor dermabrader till pinpoint bleeding was seen. The paste-like material obtained by this procedure containing melanocytes and keratinocytes admixed with the ointment base was harvested with spatula and was subsequently spread over the recipient area. Recipient site was prepared in the same manner by dermabrasion. After 10 days, dressing at both sites was removed taking utmost care at the recipient site as there was a theoretical risk of dislodging epidermal cells. Results: In a study of 437 vitiligo patches, more than 75% re-pigmentation (excellent improvement was seen in 41% of the patches. Lesions on thigh (100%, face (75% and trunk (50% showed maximal excellent improvement, whereas patches on joints and acral areas did not show much improvement. Conclusions: This technique is a simplified, cost effective, less time-consuming alternative to other techniques which involve tryspsinisation of melanocytes and at the same time provides satisfactory uniform pigmentation.

  4. Evaluation of culture-based techniques and 454 pyrosequencing for the analysis of fungal diversity in potting media and organic fertilizers.

    Science.gov (United States)

    Al-Sadi, A M; Al-Mazroui, S S; Phillips, A J L

    2015-08-01

    Potting media and organic fertilizers (OFs) are commonly used in agricultural systems. However, there is a lack of studies on the efficiency of culture-based techniques in assessing the level of fungal diversity in these products. A study was conducted to investigate the efficiency of seven culture-based techniques and pyrosequencing for characterizing fungal diversity in potting media and OFs. Fungal diversity was evaluated using serial dilution, direct plating and baiting with carrot slices, potato slices, radish seeds, cucumber seeds and cucumber cotyledons. Identity of all the isolates was confirmed on the basis of the internal transcribed spacer region of the ribosomal RNA (ITS rRNA) sequence data. The direct plating technique was found to be superior over other culture-based techniques in the number of fungal species detected. It was also found to be simple and the least time consuming technique. Comparing the efficiency of direct plating with 454 pyrosequencing revealed that pyrosequencing detected 12 and 15 times more fungal species from potting media and OFs respectively. Analysis revealed that there were differences between potting media and OFs in the dominant phyla, classes, orders, families, genera and species detected. Zygomycota (52%) and Chytridiomycota (60%) were the predominant phyla in potting media and OFs respectively. The superiority of pyrosequencing over cultural methods could be related to the ability to detect obligate fungi, slow growing fungi and fungi that exist at low population densities. The evaluated methods in this study, especially direct plating and pyrosequencing, may be used as tools to help detect and reduce movement of unwanted fungi between countries and regions. © 2015 The Society for Applied Microbiology.

  5. Nuclear techniques in plant pathology. 2. Use of radioauthography in phytopathology

    International Nuclear Information System (INIS)

    Silva, D.M.; Lima Nogueira, N. de

    1986-01-01

    Through association of radioautography techniques and electron or optic microscopy were studied: the sites of virus nucleic acids replication in the tissues plants and protoplasts; interaction bacteriophage-phytopathogenic bacteria; biological functions of Hemileia-coffee plant system. The quantative radioautography allowed distinguish the source of silver grains on two or more organelles and the Southern-Blot technique, with radioative DNA has been used to find the relationship among viruses or plasmids. (Author) [pt

  6. Culture-Independent Techniques for Rapid Detection of Bacteria Associated with Loss of Chloramine Residual in a Drinking Water System

    Science.gov (United States)

    Hoefel, Daniel; Monis, Paul T.; Grooby, Warwick L.; Andrews, Stuart; Saint, Christopher P.

    2005-01-01

    Chloramination is often the disinfection regimen of choice for extended drinking water systems. However, this process is prone to instability due to the growth of nitrifying bacteria. This is the first study to use alternative approaches for rapid investigation of chloraminated drinking water system instability in which flow cytometric cell sorting of bacteria with intact membranes (membrane-intact fraction) (BacLight kit) or with active esterases (esterase-active fraction) (carboxyfluorescein diacetate) was combined with 16S rRNA gene-directed PCR and denaturing gradient gel electrophoresis (DGGE). No active bacteria were detected when water left the water treatment plant (WTP), but 12 km downstream the chloramine residual had diminished and the level of active bacteria in the bulk water had increased to more than 1 × 105 bacteria ml−1. The bacterial diversity in the system was represented by six major DGGE bands for the membrane-intact fraction and 10 major DGGE bands for the esterase-active fraction. PCR targeting of the 16S rRNA gene of chemolithotrophic ammonia-oxidizing bacteria (AOB) and subsequent DGGE and DNA sequence analysis revealed the presence of an active Nitrosospira-related species and Nitrosomonas cryotolerans in the system, but no AOB were detected in the associated WTP. The abundance of active AOB was then determined by quantitative real-time PCR (qPCR) targeting the amoA gene; 3.43 × 103 active AOB ml−1 were detected in the membrane-intact fraction, and 1.40 × 104 active AOB ml−1 were detected in the esterase-active fraction. These values were several orders of magnitude greater than the 2.5 AOB ml−1 detected using a routine liquid most-probable-number assay. Culture-independent techniques described here, in combination with existing chemical indicators, should allow the water industry to obtain more comprehensive data with which to make informed decisions regarding remedial action that may be required either prior to or during an

  7. Amino acid transport across the tonoplast of vacuoles isolated from barley mesophyll protoplasts: Uptake of alanine, leucine, and glutamine

    International Nuclear Information System (INIS)

    Dietz, K.J.; Jaeger, R.; Kaiser, G.; Martinoia, E.

    1990-01-01

    Mesophyll protoplasts from leaves of well-fertilized barley (Hordeum vulgare L.) plants contained amino acids at concentrations as high as 120 millimoles per liter. With the exception of glutamic acid, which is predominantly localized in the cytoplasm, a major part of all other amino acids was contained inside the large central vacuole. Alanine, leucine, and glutamine are the dominant vacuolar amino acids in barley. Their transport into isolated vacuoles was studied using 14 C-labeled amino acids. Uptake was slow in the absence of ATP. A three- to sixfold stimulation of uptake was observed after addition of ATP or adenylyl imidodiphosphate an ATP analogue not being hydrolyzed by ATPases. Other nucleotides were ineffective in increasing the rate of uptake. ATP-Stimulated amino acid transport was not dependent on the transtonoplast pH or membrane potential. p-Chloromercuriphenylsulfonic acid and n-ethyl maleimide increased transport independently of ATP. Neutral amino acids such as valine or leucine effectively decreased the rate of alanine transport. Glutamine and glycine were less effective or not effective as competitive inhibitors of alanine transport. The results indicate the existence of a uniport translocator specific for neutral or basic amino acids that is under control of metabolic effectors

  8. Bioethanol production by a flocculent hybrid, CHFY0321 obtained by protoplast fusion between Saccharomyces cerevisiae and Saccharomyces bayanus

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Gi-Wook; Kang, Hyun-Woo; Kim, Yule [Changhae Institute of Cassava and Ethanol Research, Changhae Ethanol Co., LTD, Palbok-Dong 829, Dukjin-Gu, Jeonju 561-203 (Korea); Um, Hyun-Ju; Kim, Mina; Kim, Yang-Hoon [Department of Microbiology, Chungbuk National University, 410 Sungbong-Ro, Heungduk-Gu, Cheongju 361-763 (Korea)

    2010-08-15

    Fusion hybrid yeast, CHFY0321, was obtained by protoplast fusion between non-flocculent-high ethanol fermentative Saccharomyces cerevisiae CHY1011 and flocculent-low ethanol fermentative Saccharomyces bayanus KCCM12633. The hybrid yeast was used together with the parental strains to examine ethanol production in batch fermentation. Under the conditions tested, the fusion hybrid CHFY0321 flocculated to the highest degree and had the capacity to ferment well at pH 4.5 and 32 C. Simultaneous saccharification and fermentation for ethanol production was carried out using a cassava (Manihot esculenta) powder hydrolysate medium containing 19.5% (w v{sup -1}) total sugar in a 5 l lab scale jar fermenter at 32 C for 65 h with an agitation speed of 2 Hz. Under these conditions, CHFY0321 showed the highest flocculating ability and the best fermentation efficiency for ethanol production compared with those of the wild-type parent strains. CHFY0321 gave a final ethanol concentration of 89.8 {+-} 0.13 g l{sup -1}, a volumetric ethanol productivity of 1.38 {+-} 0.13 g l{sup -1} h{sup -1}, and a theoretical yield of 94.2 {+-} 1.58%. These results suggest that CHFY0321 exhibited the fermentation characteristics of S. cerevisiae CHY1011 and the flocculent ability of S. bayanus KCCM12633. Therefore, the strong highly flocculent ethanol fermentative CHFY0321 has potential for improving biotechnological ethanol fermentation processes. (author)

  9. Bioethanol production by a flocculent hybrid, CHFY0321 obtained by protoplast fusion between Saccharomyces cerevisiae and Saccharomyces bayanus

    International Nuclear Information System (INIS)

    Choi, Gi-Wook; Um, Hyun-Ju; Kang, Hyun-Woo; Kim, Yule; Kim, Mina; Kim, Yang-Hoon

    2010-01-01

    Fusion hybrid yeast, CHFY0321, was obtained by protoplast fusion between non-flocculent-high ethanol fermentative Saccharomyces cerevisiae CHY1011 and flocculent-low ethanol fermentative Saccharomyces bayanus KCCM12633. The hybrid yeast was used together with the parental strains to examine ethanol production in batch fermentation. Under the conditions tested, the fusion hybrid CHFY0321 flocculated to the highest degree and had the capacity to ferment well at pH 4.5 and 32 o C. Simultaneous saccharification and fermentation for ethanol production was carried out using a cassava (Manihot esculenta) powder hydrolysate medium containing 19.5% (w v -1 ) total sugar in a 5 l lab scale jar fermenter at 32 o C for 65 h with an agitation speed of 2 Hz. Under these conditions, CHFY0321 showed the highest flocculating ability and the best fermentation efficiency for ethanol production compared with those of the wild-type parent strains. CHFY0321 gave a final ethanol concentration of 89.8 ± 0.13 g l -1 , a volumetric ethanol productivity of 1.38 ± 0.13 g l -1 h -1 , and a theoretical yield of 94.2 ± 1.58%. These results suggest that CHFY0321 exhibited the fermentation characteristics of S. cerevisiae CHY1011 and the flocculent ability of S. bayanus KCCM12633. Therefore, the strong highly flocculent ethanol fermentative CHFY0321 has potential for improving biotechnological ethanol fermentation processes.

  10. Radiosensitivity of Nicotiana protoplasts. Action on cell; cycle effects of low dose and fractionated irradiations; biological repair

    International Nuclear Information System (INIS)

    Magnien, E.

    1981-10-01

    Leaf protoplasts of Nicotiana plumbaginifolia and Nicotiana sylvestris demonstrate five main qualities: they can be maintained as haploid lines; they constitute starting populations with a remarkable cytological homogeneity; they show a transient initial lag-phase; they yield very high plating efficiencies and retain permanently a complete differentiation capacity; being derived of a cell wall, they appear well adapted for fusion experiments or enzymatic dosages. The resumption of mitotic activity was followed by cytophotometric measurements, labelling experiments, nuclear sizing and enzymatic assays. The action of 5 Gy gamma-ray irradiations delayed entrance in the S-phase, provoked an otherwise not verified dependency between transcription, translation and protein synthesis, increased nuclear volumes in the G2-phase, and slightly stimulated the activity of a repair enzyme. The plating efficiency was a sensitive end-point which allowed the evaluation of the biological effectiveness of low to medium radiation-doses after gamma-ray and fast neutron irradiations. The neutron dose-RBE relationship increased from 3 to 25 when the dose decreased from 5 Gy to 5 mGy. When fractionated into low single doses only, a neutron dose of 300 mGy markedly increased its biological effectiveness: this phenomenon could not be explained by cell progression, and necessitated additional hypotheses involving other mechanisms in the specific action of low radiation doses. Radiation-induced UDS was measured in presence of aphidicolin. A beta-like DNA-polymerase was shown to be definitely involved in nuclear repair synthesis [fr

  11. Localization of viral antigens in leaf protoplasts and plants by immunogold labelling

    NARCIS (Netherlands)

    Lent, van J.W.M.

    1988-01-01

    This thesis describes the application of an immunocytochemical technique, immunogold labelling, new in the light and electron microscopic study of the plant viral infection. In Chapter 1 the present state of knowledge of the plant viral infection process, as revealed by

  12. Diagnostic accuracy of semi-quantitative and quantitative culture techniques for the diagnosis of catheter-related infections in newborns and molecular typing of isolated microorganisms.

    Science.gov (United States)

    Riboli, Danilo Flávio Moraes; Lyra, João César; Silva, Eliane Pessoa; Valadão, Luisa Leite; Bentlin, Maria Regina; Corrente, José Eduardo; Rugolo, Ligia Maria Suppo de Souza; da Cunha, Maria de Lourdes Ribeiro de Souza

    2014-05-22

    Catheter-related bloodstream infections (CR-BSIs) have become the most common cause of healthcare-associated bloodstream infections in neonatal intensive care units (ICUs). Microbiological evidence implicating catheters as the source of bloodstream infection is necessary to establish the diagnosis of CR-BSIs. Semi-quantitative culture is used to determine the presence of microorganisms on the external catheter surface, whereas quantitative culture also isolates microorganisms present inside the catheter. The main objective of this study was to determine the sensitivity and specificity of these two techniques for the diagnosis of CR-BSIs in newborns from a neonatal ICU. In addition, PFGE was used for similarity analysis of the microorganisms isolated from catheters and blood cultures. Semi-quantitative and quantitative methods were used for the culture of catheter tips obtained from newborns. Strains isolated from catheter tips and blood cultures which exhibited the same antimicrobial susceptibility profile were included in the study as positive cases of CR-BSI. PFGE of the microorganisms isolated from catheters and blood cultures was performed for similarity analysis and detection of clones in the ICU. A total of 584 catheter tips from 399 patients seen between November 2005 and June 2012 were analyzed. Twenty-nine cases of CR-BSI were confirmed. Coagulase-negative staphylococci (CoNS) were the most frequently isolated microorganisms, including S. epidermidis as the most prevalent species (65.5%), followed by S. haemolyticus (10.3%), yeasts (10.3%), K. pneumoniae (6.9%), S. aureus (3.4%), and E. coli (3.4%). The sensitivity of the semi-quantitative and quantitative techniques was 72.7% and 59.3%, respectively, and specificity was 95.7% and 94.4%. The diagnosis of CR-BSIs based on PFGE analysis of similarity between strains isolated from catheter tips and blood cultures showed 82.6% sensitivity and 100% specificity. The semi-quantitative culture method showed higher

  13. Preparation of anti-inflammatory mesenchymal stem/precursor cells (MSCs) through sphere formation using hanging-drop culture technique.

    Science.gov (United States)

    Bartosh, Thomas J; Ylostalo, Joni H

    2014-02-06

    Herein, we describe a protocol for preparation of pre-activated anti-inflammatory human mesenchymal stem/precursor cells (MSCs) in 3-D culture without addition of exogenous chemicals or gene-transfer approaches. MSCs are an easily procurable source of multipotent adult stem cells with therapeutic potential largely attributed to their paracrine regulation of inflammation and immunity. However, the culture conditions to prepare the ideal MSCs for cell therapy remain elusive. Furthermore, the reported lag time for activation in experimental models has prompted investigations on pre-activating the cells prior to their administration. In this protocol, standard 2-D culture-expanded MSCs are activated by aggregation into 3-D spheres using hanging-drop cultures. MSC activation is evaluated by real-time PCR and/or ELISA for anti-inflammatory factors (TSG-6, STC-1, PGE2), and by a functional assay using lipopolysaccharide-stimulated macrophage cultures. Further, we elucidate methods to prepare MSC-sphere conditioned medium, intact spheres, and suspension of single cells from spheres for experimental and clinical applications. Copyright © 2014 John Wiley & Sons, Inc.

  14. Preparation of anti-inflammatory mesenchymal stem/precursor cells (MSCs) through sphere formation using hanging drop culture technique

    Science.gov (United States)

    Bartosh, Thomas J.

    2014-01-01

    Herein, we describe a protocol for preparation of pre-activated anti-inflammatory human mesenchymal stem/precursor cells (MSCs) in 3D culture without addition of exogenous chemicals or gene transfer approaches. MSCs are an easily procurable source of multipotent adult stem cells with therapeutic potential largely attributed to their paracrine regulation of inflammation and immunity. However, the culture conditions to prepare the ideal MSCs for cell therapy remain elusive. Furthermore, reported lag time for activation in experimental models have prompted investigations to pre-activate the cells prior to their administration. In this protocol, standard 2D culture expanded MSCs are activated by aggregation into 3D spheres using hanging drop cultures. MSC activation is evaluated by real-time PCR and/or ELISA for anti-inflammatory factors (TSG-6, STC-1, PGE2), and by a functional assay using lipopolysaccharide-stimulated macrophage cultures. Furthermore, we elucidate methods to prepare MSC sphere conditioned medium, intact spheres, and suspension of single cells from spheres for experimental and clinical applications. PMID:24510769

  15. Tissue Culture Technique and Gamma Irradiation Used in Evaluation of Five Genotypes of Bread Wheat to Salinity Tolerance

    International Nuclear Information System (INIS)

    AL Jibouri, A. A. M.; Sulaiman, A. A.; Dallul, R. A.

    2005-01-01

    Callus has been induced from immature embryos of five genotypes of Triticum aestivum L (i.e. Maxipak, IPA99, 113, Tamus 3 and Babylon) on MS culture media. The calli were irradiated by Gamma ray at doses (0,15, 30,45, And 60) Gray, and cultured on MS media supplemented with salt mixture consist of sodium, calcium and magnesium chlorides in portions 3; 3; 1 respectively.The results showed significant differences in genotype responses to irradiation and salinity . A significant reduction in fresh and dry weights with increasing of irradiation dose and salt concentration in culture media in all genotypes. The genotype IPA99 showed a highest fresh and dry weight than the other genotypes . The proline contents increased significantly in callus with irradiation doses and concentration increases. On the other hand callus carbohydrate content reduced significantly with increased salt concentration in culture media. The capability of callus regeneration was reduced by increasing irradiation doses and salt concentration in culture media. (author)

  16. Production of mutants by irradiation of in vitro-cultured tissues of coconut and banana and their mass propagation by the tissue culture technique

    International Nuclear Information System (INIS)

    Guzman, E.V. de; Rosario, A.G. del; Pagcaliwagan, P.C.

    1982-01-01

    Regeneration of buds/shoots as well as plantlets was induced from banana shoot tip explants cultured in highly modified Murashige and Skoog's medium supplemented with coconut water and benzyladenine. Initially shoot regeneration was sparse, but on further subculture became profuse. Gamma irradiation at low dosage (1.0 kR) was stimulating to explant growth and bud formation with the two types of explants used. With Bungulan stimulation was observed even at 2.5 kR. Several morphological aberrations were exhibited by shoots of 'irradiated' in vitro plants growing in potted soil. A highly and continuously proliferating tissue strain has been isolated from a subculture which was ultimately derived from an irradiated explant. Its continued proliferation is dependent on an external supply of coconut water and benzyladenine. In vitro-produced plants have been established under field conditions. The 'irradiated' plants are comparable with, and some seem to be better than, the unirradiated controls with respect to height, girth, sucker production and number of hands and fingers per bunch. Higher doses of irradiation are required to produce an adverse effect on growth of coconut embryos during the liquid culture than when growing in solid medium. (author)

  17. Application of aesthetic, recreational forms of physical culture in the organizational and pedagogical techniques of school physical education

    Directory of Open Access Journals (Sweden)

    Roters T.T.

    2010-06-01

    Full Text Available The results of methodological analysis are presented in relation to introduction of organizational pedagogical technologies of leadthrough of the employments fixed, extracurricular and extracurricular. Directions of increase of interest and motivation of schoolboys are rotined to employments by physical exercises with the use of aesthetically beautiful forms of physical culture. It is indicated that modern organizational pedagogical technologies come forward the determinative of satisfaction of needs and interests of schoolboys. It is set that the aesthetic recreation forms of physical culture render assistance to the increase of interest and motivation to employments by physical exercises.

  18. Microbial diversity and dynamics throughout manufacturing and ripening of surface ripened semi-hard Danish Danbo cheeses investigated by culture-independent techniques.

    Science.gov (United States)

    Ryssel, Mia; Johansen, Pernille; Al-Soud, Waleed Abu; Sørensen, Søren; Arneborg, Nils; Jespersen, Lene

    2015-12-23

    Microbial successions on the surface and in the interior of surface ripened semi-hard Danish Danbo cheeses were investigated by culture-dependent and -independent techniques. Culture-independent detection of microorganisms was obtained by denaturing gradient gel electrophoresis (DGGE) and pyrosequencing, using amplicons of 16S and 26S rRNA genes for prokaryotes and eukaryotes, respectively. With minor exceptions, the results from the culture-independent analyses correlated to the culture-dependent plating results. Even though the predominant microorganisms detected with the two culture-independent techniques correlated, a higher number of genera were detected by pyrosequencing compared to DGGE. Additionally, minor parts of the microbiota, i.e. comprising surface and the interior of the cheeses diverged. During cheese production pyrosequencing determined Lactococcus as the dominating genus on cheese surfaces, representing on average 94.7%±2.1% of the OTUs. At day 6 Lactococcus spp. declined to 10.0% of the OTUs, whereas Staphylococcus spp. went from 0.0% during cheese production to 75.5% of the OTUs at smearing. During ripening, i.e. from 4 to 18 weeks, Corynebacterium was the dominant genus on the cheese surface (55.1%±9.8% of the OTUs), with Staphylococcus (17.9%±11.2% of the OTUs) and Brevibacterium (10.4%±8.3% of the OTUs) being the second and third most abundant genera. Other detected bacterial genera included Clostridiisalibacter (5.0%±4.0% of the OTUs), as well as Pseudoclavibacter, Alkalibacterium and Marinilactibacillus, which represented surface ripened semi-hard cheeses. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. The plant tissue culture

    International Nuclear Information System (INIS)

    Crocomo, O.J.; Sharp, W.R.

    1973-01-01

    Progress in the field of plant tissue culture at the Plant Biochemistry Sector, Centro de Energia na Agricultura (CENA), Piracicaba, S.P., Brazil, pertains to the simplification of development in 'Phaseolus vulgaris' by dividing the organism into its component organs, tissues, and cells and the maintenance of these components on defined culture media 'in vitro'. This achievement has set the stage for probing the basis for the stability of the differentiated states and/or the reentry of mature differentiated cells into the mitotic cell cycle and their subsequent redifferentiation. Data from such studies at the cytological and biochemical level have been invaluable in the elucidation of the control mechanisms responsible for expression of the cellular phenotype. Unlimited possibilities exist for the application of tissue culture in the vegetative propagation of 'Phaseolus' and other important cultivars in providing genocopies or a large scale and/or readily obtaining plantlets from haploid cell lines or from protoplast (wall-less cells) hybridization products following genetic manipulation. These tools are being applied in this laboratory for the development and selection of high protein synthesizing 'Phaseolus' cultivars

  20. Determination of the cause of the symptoms on yellow yam (Dioscorea cayenensis Lam.) leaf tissue and their eradication, enriching the culture medium and using techniques of meristem culture, thermo and chemotherapy on in vitro conditions

    International Nuclear Information System (INIS)

    Brenes Huertas, Mauricio

    2010-01-01

    Yams (Dioscorea spp) has been cultivated for exportation in Costa Rica, in North Huetar region. In vitro culture technique has been used for multiplying planting material for many advantages. However, cleaning of viruses that affect has been ineffective. Viruses such as: the potyvirus, potexvirus, cucumovirus . Methods like meristem culture, chemotherapy, thermotherapy and combinations of these have been used for the elimination of virus in plant species. The plants were evaluated in indexing assays, observing symptoms, serological methods and electron microscopy, among others. Other problems that have been affecting in vitro plant are deficient culture media in some nutrient. The presence of some abnormal characteristics in leaf tissue was determined whether have been caused by a virus or a nutritional deficiency in the culture medium. The presence of the virus has tried to find using ELISA and electron microscopy. Tests meristem culture, thermotherapy and chemotherapy have been made for the eradication of a possible virus; which have been assessed by observation of symptomatology and ELISA. The efficiency of the culture medium was evaluated to enrich it with nitrogen or excess iron. None of the suspected virus found in ELISA tests. Filaments are presumably viral particles were found through analysis of ultrastructure, as well as alterations in chloroplasts which indicated the presence of a pathogen or toxicity. Thermotherapy and chemotherapy with the concentration of 40 mg/L of ribavirin have been the most effective for the elimination of symptoms in virus eradication treatments. Assessments nutrient concentrations have shown that the differences between the various treatments used were undetectable. The symptoms presented were caused, according to the conclusions, by a virus which should preferably deal with thermotherapy. (author) [es

  1. Production of solid mutants in citrus, utilizing new approaches and techniques

    International Nuclear Information System (INIS)

    Spiegel-Roy, P.; Kochba, J.

    1975-01-01

    Conditions for embryoid differentiation in Shamouti orange ovular callus were studied. Lines differing in embryogenic capacity were established. Ageing of callus prior to subculture enhanced embryoid development. A pronounced habituation effect has been found in most cultures. Protoplasts have been obtained from callus after treatment with cellolytic enzymes. Irradiation of any embryogenic line of callus with 12-20 kR seemed to promote embryoid formation, after a time lag in their appearance. Transferrence of embryoids into agar+sucrose and GA 3 and a further transfer into agar+sucrose, GA 3 and adenine sulphate gave best results as to rooting of embryoids and plant survival. The technique of using decapitated nucellar seedlings for mutagenic treatment was developed further with Shamouti orange and Marsh grapefruit. Irradiation 48-72 hours after decapitation with 2 kR resulted in shoot neoformation and survival not much below that of control, while 6 kR impeded de novo shoot formation in Marsh grapefruit. A tendency towards mutants with earlier fruit maturity was found in mV 2 plants from material originating from irradiation with the 8-kR dose. (author)

  2. 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

  3. Naturally induced secretions of the potato cyst nematode co-stimulate the proliferation of both tobacco leaf protoplasts and human peripheral blood mononuclear cells.

    Science.gov (United States)

    Goverse, A; Rouppe van der Voort, J; Roppe van der Voort, C; Kavelaars, A; Smant, G; Schots, A; Bakker, J; Helder, J

    1999-10-01

    Naturally induced secretions from infective juveniles of the potato cyst nematode Globodera rostochiensis co-stimulate the proliferation of tobacco leaf protoplasts in the presence of the synthetic phytohormones alpha-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP). With the use of a protoplast-based bioassay, a low-molecular-weight peptide(s) (cyst nematode secretions also co-stimulated mitogenesis in human peripheral blood mononuclear cells (PBMC). The stimulation of plant cells isolated from nontarget tissue--these nematodes normally invade the roots of potato plants--suggests the activation of a general signal transduction mechanism(s) by an oligopeptide(s) secreted by the nematode. Whether a similar oligopeptide-induced mechanism underlies human PBMC activation remains to be investigated. Reactivation of the cell cycle is a crucial event in feeding cell formation by cyst nematodes. The secretion of a mitogenic low-molecular-weight peptide(s) by infective juveniles of the potato cyst nematode could contribute to the redifferentiation of plant cells into such a feeding cell.

  4. Effects of different antibiotic classes on airway bacteria in stable COPD using culture and molecular techniques: a randomised controlled trial

    Science.gov (United States)

    Brill, Simon E; Law, Martin; El-Emir, Ethaar; Allinson, James P; James, Phillip; Maddox, Victoria; Donaldson, Gavin C; McHugh, Timothy D; Cookson, William O; Moffatt, Miriam F; Nazareth, Irwin; Hurst, John R; Calverley, Peter M A; Sweeting, Michael J; Wedzicha, Jadwiga A

    2015-01-01

    Background Long-term antibiotic therapy is used to prevent exacerbations of COPD but there is uncertainty over whether this reduces airway bacteria. The optimum antibiotic choice remains unknown. We conducted an exploratory trial in stable patients with COPD comparing three antibiotic regimens against placebo. Methods This was a single-centre, single-blind, randomised placebo-controlled trial. Patients aged ≥45 years with COPD, FEV1<80% predicted and chronic productive cough were randomised to receive either moxifloxacin 400 mg daily for 5 days every 4 weeks, doxycycline 100 mg/day, azithromycin 250 mg 3 times a week or one placebo tablet daily for 13 weeks. The primary outcome was the change in total cultured bacterial load in sputum from baseline; secondary outcomes included bacterial load by 16S quantitative PCR (qPCR), sputum inflammation and antibiotic resistance. Results 99 patients were randomised; 86 completed follow-up, were able to expectorate sputum and were analysed. After adjustment, there was a non-significant reduction in bacterial load of 0.42 log10 cfu/mL (95% CI −0.08 to 0.91, p=0.10) with moxifloxacin, 0.11 (−0.33 to 0.55, p=0.62) with doxycycline and 0.08 (−0.38 to 0.54, p=0.73) with azithromycin from placebo, respectively. There were also no significant changes in bacterial load measured by 16S qPCR or in airway inflammation. More treatment-related adverse events occurred with moxifloxacin. Of note, mean inhibitory concentrations of cultured isolates increased by at least three times over placebo in all treatment arms. Conclusions Total airway bacterial load did not decrease significantly after 3 months of antibiotic therapy. Large increases in antibiotic resistance were seen in all treatment groups and this has important implications for future studies. Trial registration number clinicaltrials.gov (NCT01398072). PMID:26179246

  5. Historical and Epistemological Reflections on the Culture of Machines around the Renaissance: How Science and Technique Work?

    Directory of Open Access Journals (Sweden)

    Raffaele Pisano

    2014-10-01

    Full Text Available This paper is divided into two parts, this being the first one. The second is entitled ‘Historical and Epistemological Reflections on the Culture of Machines around Renaissance: Machines, Machineries and Perpetual Motion’ and will be published in Acta Baltica Historiae et Philosophiae Scientiarum in 2015. Based on our recent studies, we provide here a historical and epistemological feature on the role played by machines and machineries. Ours is an epistemological thesis based on a series of historical examples to show that the relations between theoretical science and the construction of machines cannot be taken for granted, a priori. Our analysis is mainly based on the culture of machines around 15th and 17th centuries, namely the epoch of Late Renaissance and Early Modern Age. For this is the period of scientific revolution and this age offers abundant interesting material for researches into the relations of theoretical science/construction of machines as well. However, to prove our epistemological thesis, we will also exploit examples of machines built in other historical periods. Particularly, a discussion concerning the relationship between science theory and the development of science art crafts produced by non-recognized scientists in a certain historical time is presented. The main questions are: when and why did the tension between science (physics, mathematics and geometry give rise to a new scientific approach to applied discipline such as studies on machines and machineries? What kind of science was used (if at all for projecting machines and machineries? Was science at the time a necessary precondition to build a machine? In the first part we will focus on the difference between Aristotelian-Euclidean and Archimedean approaches and we will outline the heritage of these two different approaches in late medieval and Renaissance science. In the second part, we will apply our reconstructions to some historical and epistemological

  6. Microspores irradiation in anther culture: testing a new technique to obtain mutations immediatly detected and fixed (Application to Nicotiana tabacum)

    International Nuclear Information System (INIS)

    Mondeil, Fanja

    1974-01-01

    In order to consider the effects of microspores irradiation on embryo development, and in order to observe the morphological responses of haploid plantlets derived from androgenetic anthers to ionizing irradiation, 1000, 1500 and 2000r of gamma rays were delivered on anthers of Nicotiana tabacum (DL 50 range calculated: 1500r). The cytological studies of embryo development revealed an apparent increase in irradiated microspores: cell division is stimulated but followed by an early mortality. A sharp rise in lethality effects was observed when gamma rays were applied beyond the seventh day of culture, when the proembryo contains an average of 4 cells. Morphological aberrations and colour changes in the Mo progeny derived from irradiated microspores are diverse. But after chromosome doubling and mutation checking out, all the plants were not recorded to have transmitted their aberrant characters. Thus, heritable character 'mutations) and not heritable character (variations) were obtained. The variations characters include dwarfing, excessive branching, fasciation and dichotomy of the stems, altered flower form, especially of petals. As to the leaves, they usually show induced changes in their colour (chlorotic areas, mosaic-colour changes, or an over-all colour changes), in their form (irregularity in outline) and in their texture (thickening, hairless leaf). Among the mutants, a monster tobacco, with excrescences on the leaves and the flowers is certainly the most conspicuous. But mutants also include altered leaf colour (over-all pale green) and altered flower colour, (dark red, clear pink, white) [fr

  7. CULTURAL HERITAGE DOCUMENTATION AND INTEGRATED GEOMATICS TECHNIQUES IN AN EDUCATIONAL CONTEXT: CASE BOIS-DU-LUC (BELGIUM

    Directory of Open Access Journals (Sweden)

    C. Stal

    2013-07-01

    Full Text Available The Walloon Region in Belgium played a leading role in the world economy and in the industrial sciences and engineering since the 19th century. Several relicts, such as important industrial buildings or sites, are spread over a large area and are still dominating the current landscape. Some of these remnants are preserved as monuments, representing the industrial, cultural and ecological transition of the region during the last two centuries. Since 2012, UNESCO recognized the importance of four of the best preserved 19th and 20th century coal mining sites and classified them as World Heritage ("Major Mining Sites of Wallonia", Le Grand-Hornu, Bois-du- Luc, Le Bois du Cazier and Blegny Mine. The four recognised mining sites together form a strip of about 170 km long, containing important examples of the so-called 'neo classical architecture’ from the early periods of the industrial era in continental Europe. Motivated by this recognition and by the huge touristic and educational potential of the sites, a partnership between the Walloon government and Ghent University was organized. In this first stage, the collaboration focuses on the site of Bois-du-Luc in the municipality of La Louvière, containing a large series of dwellings, several equipments built between 1853 and 1923 by the Société des Charbonnages du Bois-du-Luc, workshops and heaps. Consisting of numerous buildings dating back to the period between 1838 and 1923, this site is one of Europe’s oldest collieries. The collaboration between the Walloon government and Ghent University – Department of Geography fits in the hands-on training which students in land surveying and geomatics receive during their Bachelor studies. For the students it is very interesting that their practical exercises are not limited to the university campus, but that they are involved in a real measurement campaign. The project consists of a series of intensive land surveying campaigns, resulting in highly

  8. The Spiritual Form of Ancient Art and Culture - Bharatanatyam (Visual Art Depicted Using Unique Techniques on Scratchboard (Fine Art Medium

    Directory of Open Access Journals (Sweden)

    Arpitha Parthasarathy

    2017-03-01

    Full Text Available The most ancient form of dance that is prevailing todays is a form of classical Indian dance, Bharatanatyam. In Sanskrit (and Devanagri, bharatanatyam means "Indian dance", is believed to have divine origin and is of the most ancient form of classical dance. Bharatanatyam is a two thousand-year-old dance form, originally practiced in the temples of ancient India. The art today remains purely devotional even today and this performing art is yet to gain awareness and interest in the western world. This dance form has various implications in improving the higher order thinking in children and provides health benefits in adults apart from cultural preservation. The current study uses scratchboard as a medium to display the artistic movements and emotions. Scratchboard, a fine art is one means by which the visual art is expressed in this current study using sharp tools, namely X-acto 11 scalpel and tattoo needles. This unique medium made up of a masonite hardboard coated with soft clay and Indian ink has been used to not only show the details of the ancient dance form and expression but also to comprehend and transcribe both visual art and fine art. It is for the first time that scratchboard medium has been the innovatively used to show various textures of flower, glistening gold jewels, hand woven silk and the divine expression in the same art ‘devotion’. The current study was carried out in-order to perpetuate, conserve and disseminate these classic forms of visual art and fine art.

  9. Application of mutation breeding technique for producing NaCl tolerant plants of banana in tissue culture and greenhouse conditions

    International Nuclear Information System (INIS)

    Vedadi, C.; Rahimi, M.; Naserian, B.; Rahmani, E.; Neshan, N.

    2005-01-01

    Full text: To study of possibility to induce salt tolerant clones in banana by using mutation technique, an experiment was conducted with factorial (gamma irradiation and salt concentration factors) in a CRD design. In this research, plantlets of banana cv. Dwarf Cavendish were produced by subculture of irradiated shoot tips. It deserves to mention that consequent subculturing was aimed at getting rid of chimera. Next, these explants were transferred to MS medium containing 2.5 mg. l- 1 BAP and NaCl concentrations of 0, 6, 7, 8, 9 g.l -1 for 2 months .Then, living buds were transferred to medium without salt. After one month, we repeated the first stage. All living buds rooted and were transferred to potted soil. Acclimatized plants were irrigated weekly with above NaCl solution. Other irrigation was done with salt-free water. There was also a negative relation between salt concentration and survival - proliferation. In second salinity stress, salt had no significant difference on survival percentage. No-significant difference of effect salt on survival in second salinity stress was observed. (author)

  10. Prenatal diagnosis of congenital toxoplasmosis: comparative value of fetal blood and amniotic fluid using serological techniques and cultures.

    Science.gov (United States)

    Fricker-Hidalgo, H; Pelloux, H; Muet, F; Racinet, C; Bost, M; Goullier-Fleuret, A; Ambroise-Thomas, P

    1997-09-01

    The prenatal diagnosis of congenital toxoplasmosis is mainly based on biological tests performed on fetal blood and amniotic fluid. We studied the performance of neonatal diagnosis procedures and the results of fetal blood and amniotic fluid analysis. Of 127 women who contracted toxoplasmosis and underwent prenatal diagnosis, the postnatal serological follow-up was long enough to definitively diagnose congenital toxoplasmosis in 19 cases and to exclude it in 27 cases. Prenatal diagnosis allowed the detection of 94.7 per cent (18/19) of the infected fetuses. The sensitivities of tests in amniotic fluid and fetal blood were equivalent, 88.2 per cent (15/17) and 87.5 per cent (14/16), respectively. In fetal blood, biological techniques were positive in 12/16 cases and in 2/16 cases, serological tests were the only positive sign. The specificities of tests in amniotic fluid and fetal blood were respectively 100 per cent (23/23) and 86.3 per cent (19/22) (three false-positive serological results). These results, added to the lower morbidity of amniocentesis compared with cordocentesis, might lead to cordocentesis being abandoned in the prenatal diagnosis of congenital toxoplasmosis.

  11. Proton induced X-ray emission (PIXE) technique for determining multi-element composition of transformed hairy root cultures of Boerhaavia diffusa L. An important medicinal herb

    International Nuclear Information System (INIS)

    Lopamudra Sahu; Chand, P.K.; Ray, D.K.; Utkal University, Vani Vihar, Bhubaneswar, Odisha

    2014-01-01

    Therapeutically important inorganic elements in Agrobacterium rhizogenes-mediated genetically transformed hairy root cultures (HRCs) of a pharmaceutically significant herb Boerhaavia diffusa were quantified using proton induced X-ray emission technique. This was compared with that of roots from the naturally grown donor plant. Two macro-elements (Ca and K) and eight different trace elements namely V, Cr, Mn, Fe, Co, Cu Zn, and Ni were detected and their content was determined. In HRCs of a transformed rhizoclone, calcium and potassium had values which were significantly higher than that of in vivo roots. The concentrations of several trace elements, which are known to have a positive implication in human healthcare, were found to be either comparable (Fe) to that in the natural root samples or higher (Mn, Zn, Cr, Cu, Co) in the transformed rhizoclone. The genetically transformed HRCs can thus serve as a fast-proliferating renewable resource of medicinally useful minerals targeting specific diseases. (author)

  12. Characterization of the Bacterial Community Naturally Present on Commercially Grown Basil Leaves: Evaluation of Sample Preparation Prior to Culture-Independent Techniques

    Directory of Open Access Journals (Sweden)

    Siele Ceuppens

    2015-08-01

    Full Text Available Fresh herbs such as basil constitute an important food commodity worldwide. Basil provides considerable culinary and health benefits, but has also been implicated in foodborne illnesses. The naturally occurring bacterial community on basil leaves is currently unknown, so the epiphytic bacterial community was investigated using the culture-independent techniques denaturing gradient gel electrophoresis (DGGE and next-generation sequencing (NGS. Sample preparation had a major influence on the results from DGGE and NGS: Novosphingobium was the dominant genus for three different basil batches obtained by maceration of basil leaves, while washing of the leaves yielded lower numbers but more variable dominant bacterial genera including Klebsiella, Pantoea, Flavobacterium, Sphingobacterium and Pseudomonas. During storage of basil, bacterial growth and shifts in the bacterial community were observed with DGGE and NGS. Spoilage was not associated with specific bacterial groups and presumably caused by physiological tissue deterioration and visual defects, rather than by bacterial growth.

  13. Simulation and evaluation of the absorption edge subtraction technique in energy-resolved X-ray radiography applied to the cultural heritage studies

    International Nuclear Information System (INIS)

    Leyva Pernia, Diana; Cabal Rodriguez, Ana E.; Pinnera Hernandez, Ibrahin; Leyva Fabelo, Antonio; Abreu Alfonso, Yamiel; Espen, Piet Van

    2011-01-01

    In this work the mathematical simulation of photon transport in the matter was used to evaluate the potentials of a new energy-resolved X-ray radiography system. The system is intended for investigations of cultural heritage object, mainly painting. The radiographic system uses polychromatic radiation from an X-ray tube and measures the spectrum transmitted through the object with an energy-dispersive X-ray detector on a pixel-by-pixel basis. Manipulation of the data-set obtained allows constructing images with enhanced contrast for certain elements. Here the use of the absorption edge subtraction technique was emphasized. The simulated results were in good agreement with the experimental data.(author)

  14. D Models for All: Low-Cost Acquisition Through Mobile Devices in Comparison with Image Based Techniques. Potentialities and Weaknesses in Cultural Heritage Domain

    Science.gov (United States)

    Santagati, C.; Lo Turco, M.; Bocconcino, M. M.; Donato, V.; Galizia, M.

    2017-11-01

    Nowadays, 3D digital imaging proposes effective solutions for preserving the expression of human creativity across the centuries, as well as is a great tool to guarantee global dissemination of knowledge and wide access to these invaluable resources of the past. Nevertheless, in several cases, a massive digitalisation of cultural heritage items (from the archaeological site up to the monument and museum collections) could be unworkable due to the still high costs in terms of equipment and human resources: 3D acquisition technologies and the need of skilled team within cultural institutions. Therefore, it is necessary to explore new possibilities offered by growing technologies: the lower costs of these technologies as well as their attractive visual quality constitute a challenge for researchers. Besides these possibilities, it is also important to consider how information is spread through graphic representation of knowledge. The focus of this study is to explore the potentialities and weaknesses of a newly released low cost device in the cultural heritage domain, trying to understand its effective usability in museum collections. The aim of the research is to test their usability, critically analysing the final outcomes of this entry level technology in relation to the other better assessed low cost technologies for 3D scanning, such as Structure from Motion (SfM) techniques (also produced by the same device) combined with dataset generated by a professional digital camera. The final outcomes were compared in terms of quality definition, time processing and file size. The specimens of the collections of the Civic Museum Castello Ursino in Catania have been chosen as the site of experimentation.

  15. 3D MODELS FOR ALL: LOW-COST ACQUISITION THROUGH MOBILE DEVICES IN COMPARISON WITH IMAGE BASED TECHNIQUES. POTENTIALITIES AND WEAKNESSES IN CULTURAL HERITAGE DOMAIN

    Directory of Open Access Journals (Sweden)

    C. Santagati

    2017-11-01

    Full Text Available Nowadays, 3D digital imaging proposes effective solutions for preserving the expression of human creativity across the centuries, as well as is a great tool to guarantee global dissemination of knowledge and wide access to these invaluable resources of the past. Nevertheless, in several cases, a massive digitalisation of cultural heritage items (from the archaeological site up to the monument and museum collections could be unworkable due to the still high costs in terms of equipment and human resources: 3D acquisition technologies and the need of skilled team within cultural institutions. Therefore, it is necessary to explore new possibilities offered by growing technologies: the lower costs of these technologies as well as their attractive visual quality constitute a challenge for researchers. Besides these possibilities, it is also important to consider how information is spread through graphic representation of knowledge. The focus of this study is to explore the potentialities and weaknesses of a newly released low cost device in the cultural heritage domain, trying to understand its effective usability in museum collections. The aim of the research is to test their usability, critically analysing the final outcomes of this entry level technology in relation to the other better assessed low cost technologies for 3D scanning, such as Structure from Motion (SfM techniques (also produced by the same device combined with dataset generated by a professional digital camera. The final outcomes were compared in terms of quality definition, time processing and file size. The specimens of the collections of the Civic Museum Castello Ursino in Catania have been chosen as the site of experimentation.

  16. D Geomatics Techniques for AN Integrated Approach to Cultural Heritage Knowledge: the Case of San Michele in Acerboli's Church in Santarcangelo DI Romagna

    Science.gov (United States)

    Bitelli, G.; Dellapasqua, M.; Girelli, V. A.; Sanchini, E.; Tini, M. A.

    2017-05-01

    The modern Geomatics techniques, such as Terrestrial Laser Scanner (TLS) and multi-view Structure from Motion (SfM), are gaining more and more interest in the Cultural Heritage field. All the data acquired with these technologies could be stored and managed together with other information in a Historical Building Information Model (HBIM). In this paper, it will be shown the case study of the San Michele in Acerboli's church, located in Santarcangelo di Romagna, Italy. This church, dated about the 6th century A.D., represents a high relevant Romanic building of the high Medieval period. The building presents an irregular square plan with a different length of the lateral brick walls and a consequential oblique one in correspondence of the apse. Nevertheless, the different lengths of the lateral brick walls are balanced thanks to the irregular spaces between the windows. Different changes occurred during the centuries, such as the closing of the seven main doors and the building of the bell tower, in the 11th century A.D., which is nowadays the main entrance of the church. An integrated survey was realized, covering the exterior and the interior. The final 3D model represents a valid support not only for documentation, but also to maintain and manage in an integrate approach the available knowledge of this Cultural Heritage site, developing a HBIM system in which all the mentioned historical, geometrical, material matters are collected.

  17. Comparative evaluation of positive tests to Mycobacterium avium subsp. paratuberculosis in clinically healthy sheep and goats in south-west Greece using molecular techniques, serology, and culture.

    Science.gov (United States)

    Ikonomopoulos, John; Balaskas, Christos; Kantzoura, Bagia; Fragiadaki, Eirini; Pavlik, Ivo; Bartos, Milan; Lukas, John C; Gazouli, Maria

    2007-09-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the cause of paratuberculosis, which affects mainly ruminants although there is a growing concern about its possible implication in Crohn's disease in humans especially in connection with environmental spread and risks to the food chain. Retail cheese may represent a significant source of human exposure to MAP and the aim of this study was to assess MAP status in clinically healthy sheep and goats in Greece, comparing techniques routinely used in the positive diagnosis of the disease. From a total of 30 flocks, 632 sheep and goats had faecal, serum, and whole-blood samples examined by culture, complement fixation test (CFT), and polymerase chain reaction (PCR) targeted at IS900, IS1245, and IS6110. PCR produced positive results in 21% of the animals tested, with 5.6%, 3.9%, and 11.5% being identified as MAP, Mycobacterium avium subsp. avium, and Mycobacterium tuberculosis complex, respectively. CFT produced positive and suspicious results in 4.4% and 14.4% of the cases. Faecal cultures were negative in all but a single case that was identified as restriction fragment length polymorphism (RFLP)-type BC1. Agreement between results obtained by PCR and CFT was poor with isolated cases although an assessment of the MAP positive tests produced similar results for both methods. The findings indicate the need for additional measures of control, although the costs may be substantial if public health protection justifies elimination of MAP from livestock.

  18. 3D GEOMATICS TECHNIQUES FOR AN INTEGRATED APPROACH TO CULTURAL HERITAGE KNOWLEDGE: THE CASE OF SAN MICHELE IN ACERBOLI’S CHURCH IN SANTARCANGELO DI ROMAGNA

    Directory of Open Access Journals (Sweden)

    G. Bitelli

    2017-05-01

    Full Text Available The modern Geomatics techniques, such as Terrestrial Laser Scanner (TLS and multi-view Structure from Motion (SfM, are gaining more and more interest in the Cultural Heritage field. All the data acquired with these technologies could be stored and managed together with other information in a Historical Building Information Model (HBIM. In this paper, it will be shown the case study of the San Michele in Acerboli’s church, located in Santarcangelo di Romagna, Italy. This church, dated about the 6th century A.D., represents a high relevant Romanic building of the high Medieval period. The building presents an irregular square plan with a different length of the lateral brick walls and a consequential oblique one in correspondence of the apse. Nevertheless, the different lengths of the lateral brick walls are balanced thanks to the irregular spaces between the windows. Different changes occurred during the centuries, such as the closing of the seven main doors and the building of the bell tower, in the 11th century A.D., which is nowadays the main entrance of the church. An integrated survey was realized, covering the exterior and the interior. The final 3D model represents a valid support not only for documentation, but also to maintain and manage in an integrate approach the available knowledge of this Cultural Heritage site, developing a HBIM system in which all the mentioned historical, geometrical, material matters are collected.

  19. Aplicações da cultura de tecidos vegetais em fruteiras do Cerrado Applications of tissue culture techniques in Brazilian Cerrado fruit trees

    Directory of Open Access Journals (Sweden)

    Hernane Fernandes Pinhal

    2011-07-01

    Full Text Available Atualmente, percebe-se uma preocupação em relação às plantas do cerrado, com grande enfoque nas fruteiras em função de suas características e usos. Apesar de ser uma área ainda pouco explorada, é crescente o número de estudos dessas espécies nativas, dentre eles, os que abrangem as técnicas de cultura de tecidos. Isso se deve uma vez que essa ferramenta biotecnológica permite a propagação de espécies com dificuldade de germinação, minimiza o problema de sementes recalcitrantes, promove a produção de mudas em larga escala, complementa bancos de germoplasma e facilita as trocas de materiais genéticos. Dessa maneira, esta revisão visa a sumarizar o histórico e panorama atual das aplicações da cultura de tecidos em fruteiras do cerrado, proporcionando sustentação para novos estudos.Currently, it's been given a huge concern to the cerrado plants, focusing on fruit trees due to their characteristics and uses. Despite being a fairly unexplored area, the number of studies on these native species has increased, especially those involving tissue culture techniques. That's because this biotechnological tool provides the propagation of species with germination difficulty, reduces problems of recalcitrant seeds, promotes large scale seedling production, complements germplasm banks and facilitates the exchange of genetic materials. Therefore, this review summarizes the history and current situation of tissue culture techniques applied to Brazilian Cerrado fruit trees, providing support to further studies.

  20. Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. Summary progress report, May 16, 1987--June 1, 1991

    Energy Technology Data Exchange (ETDEWEB)

    Steponkus, P.L.

    1991-12-31

    This project focuses on lesions in the plasma membrane of protoplasts that occur during freezing to temperatures below {minus}5{degrees} which result in changes in the semipermeablity of the plasma membrane. This injury, referred to as loss of osmotic responsiveness, is associated with the formation of large, aparticulate domains in the plasma membrane, aparticulate lamellae subtending the plasma membrane, and lamellar-to-hexagonal{sub II} phase transitions in the plasma membrane and subtending lamellar. The goals of this project are to provide a mechanistic understanding of the mechanism by which freeze-induced dehydration effects the formation of aparticulate domains and lamellar-to-hexagonal{sub II} phase transitions and to determine the mechanisms by which cold acclimation and cryoprotectants preclude or diminish these ultrastructural changes. Our working hypothesis is the formation of aparticulate domains and lamellar-to-hexagon{sub II} phase transitions in the plasma membrane and subtending lamellae are manifestations of hydration-dependent bilayer-bilayer interactions.

  1. Hydroponic Culture

    Science.gov (United States)

    Steucek, G. L.; Yurkiewicz, W. J.

    1973-01-01

    Describes a hydroponic culture technique suitable for student exercises in biology. This technique of growing plants in nutrient solutions enhances plant growth, and is an excellent way to obtain intact plants with root systems free of soil or other particulate matter. (JR)

  2. Technical paper on the realisation of a continuous axenic culture of chlorella. Description of a biostat; Note technique sur la realisation d'une culture continue et axenique de chlorelles. Description d'un biostat

    Energy Technology Data Exchange (ETDEWEB)

    Dalmon, J.; Gilet, R. [Commissariat a l' Energie Atomique, Grenoble (France). Centre d' Etudes Nucleaires

    1966-07-01

    An apparatus for the continuous cultivation of chlorella is described with the means that allowed to keep axenic cultures for several months. (authors) [French] On decrit un appareil assurant une culture continue de chlorelles en insistant sur les modalites qui permettent de garder cette culture axenique durant plusieurs mois. (auteurs)

  3. Technical paper on the realisation of a continuous axenic culture of chlorella. Description of a biostat; Note technique sur la realisation d'une culture continue et axenique de chlorelles. Description d'un biostat

    Energy Technology Data Exchange (ETDEWEB)

    Dalmon, J; Gilet, R [Commissariat a l' Energie Atomique, Grenoble (France). Centre d' Etudes Nucleaires

    1966-07-01

    An apparatus for the continuous cultivation of chlorella is described with the means that allowed to keep axenic cultures for several months. (authors) [French] On decrit un appareil assurant une culture continue de chlorelles en insistant sur les modalites qui permettent de garder cette culture axenique durant plusieurs mois. (auteurs)

  4. Bacterial community structure associated with white band disease in the elkhorn coral Acropora palmata determined using culture-independent 16S rRNA techniques.

    Science.gov (United States)

    Pantos, Olga; Bythell, John C

    2006-03-23

    Culture-independent molecular (16S ribosomal RNA) techniques showed distinct differences in bacterial communities associated with white band disease (WBD) Type I and healthy elkhorn coral Acropora palmata. Differences were apparent at all levels, with a greater diversity present in tissues of diseased colonies. The bacterial community associated with remote, non-diseased coral was distinct from the apparently healthy tissues of infected corals several cm from the disease lesion. This demonstrates a whole-organism effect from what appears to be a localised disease lesion, an effect that has also been recently demonstrated in white plague-like disease in star coral Montastraea annularis. The pattern of bacterial community structure changes was similar to that recently demonstrated for white plague-like disease and black band disease. Some of the changes are likely to be explained by the colonisation of dead and degrading tissues by a micro-heterotroph community adapted to the decomposition of coral tissues. However, specific ribosomal types that are absent from healthy tissues appear consistently in all samples of each of the diseases. These ribotypes are closely related members of a group of alpha-proteobacteria that cause disease, notably juvenile oyster disease, in other marine organisms. It is clearly important that members of this group are isolated for challenge experiments to determine their role in the diseases.

  5. The r.b.e. of different-energy neutrons as determined by human bone-marrow cell-culture techniques

    International Nuclear Information System (INIS)

    Boeyum, A.; Carsten, A.L.; Chikkappa, G.; Cook, L.; Bullis, J.; Honikel, L.; Cronkite, E.P.

    1978-01-01

    The effect of X-rays and different-energy neutrons on human bone-marrow cells was studied using two different cell-culture techniques - diffusion chamber (DC) growth and colony formation in vitro (CFU-C). Based on the survival and proliferative granulocytes in DC on day 13, the D 0 value was 80 rad with X-rays, and 117 rad as measured by the CFU-C assay. The D 0 values for neutrons depended on the radiation source and the energy level. The r.b.e. values, which dropped with increasing energy levels of mono-energetic neutrons, were (i) 0.44 MeV; DC 3.7, CFU-C 4.1; (ii) 6 MeV; DC 1.8, CFU-C 2.0; (iii) 15 MeV; DC 1.6, CFU-C 1.6; (iv) fission neutrons; DC 2.6, CFU-C 2.4. (author)

  6. Microbial diversity and dynamics throughout manufacturing and ripening of surface ripened semi-hard Danish Danbo cheeses investigated by culture-independent techniques

    DEFF Research Database (Denmark)

    Ryssel, Mia; Johansen, Pernille; Abu Al-Soud, Waleed

    2015-01-01

    ) and pyrosequencing, using amplicons of 16S and 26S rRNA genes for prokaryotes and eukaryotes, respectively. With minor exceptions, the results from the culture-independent analyses correlated to the culture-dependent plating results. Even though the predominant microorganisms detected with the two culture...

  7. Technique of the `in vitro` fertilization and the culture of mouse embryos at preimplantation; Tecnica de fertilizacao `in vitro` e cultura de embrioes de camundongo durante a pre-implantacao

    Energy Technology Data Exchange (ETDEWEB)

    Kikuchi, Olivia Kimiko [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil); Yamada, Takeshi [National Inst. of Radiological Sciences, Chiba (Japan)

    1993-03-01

    The mammal embryo is an intensive cellular proliferating system, very radiosensitive and therefore adequate to the study of the biological effects of ionizing radiation. The technique of the in vitro fertilization and the culture of mouse embryos at preimplantation period, modified by Yamada et al (1982) to improve the efficiency of more than 95% of blastocyst formation is described. (author) 2 refs., 7 figs.

  8. Plant Tissue Culture

    Indian Academy of Sciences (India)

    Admin

    Plant tissue culture is a technique of culturing plant cells, tissues and organs on ... working methods (Box 2) and discovery of the need for B vita- mins and auxins for ... Kotte (Germany) reported some success with growing isolated root tips.

  9. Analysis of bacterial flora associated with peri-implantitis using obligate anaerobic culture technique and 16S rDNA gene sequence.

    Science.gov (United States)

    Tamura, Naoki; Ochi, Morio; Miyakawa, Hiroshi; Nakazawa, Futoshi

    2013-01-01

    To analyze and characterize the predominant bacterial flora associated with peri-implantitis by using culture techniques under obligate anaerobic conditions and 16S rDNA gene sequences. Subgingival bacterial specimens were taken from 30 patients: control (n = 15), consisting of patients with only healthy implants; and test (n = 15), consisting of patients with peri-implantitis. In both groups, subgingival bacterial specimens were taken from the deepest sites. An anaerobic glove box system was used to cultivate bacterial strains. The bacterial strains were identified by 16S rDNA genebased polymerase chain reaction and comparison of the gene sequences. Peri-implantitis sites had approximately 10-fold higher mean colony forming units (per milliliter) than healthy implant sites. A total of 69 different bacterial species were identified in the peri-implantitis sites and 53 in the healthy implant sites. The predominant bacterial species in the peri-implantitis sites were Eubacterium nodatum, E. brachy, E. saphenum, Filifactor alocis, Slackia exigua, Parascardovia denticolens, Prevotella intermedia, Fusobacterium nucleatum, Porphyromonas gingivalis, Centipeda periodontii, and Parvimonas micra. The predominant bacteria in healthy implant sites apart from Streptococcus were Pseudoramibacter alactolyticus, Veillonella species, Actinomyces israelii, Actinomyces species, Propionibacterium acnes, and Parvimonas micra. These results suggest that the environment in the depths of the sulcus showing peri-implantitis is well suited for growth of obligate anaerobic bacteria. The present study demonstrated that the sulcus around oral implants with peri-implantitis harbors high levels of asaccharolytic anaerobic gram-positive rods (AAGPRs) such as E. nodatum, E. brachy, E. saphenum, Filifactor alocis, Slackia exigua, and gram-negative anaerobic rods, suggesting that conventional periodontopathic bacteria are not the only periodontal pathogens active in peri-implantitis, and that AAGPRs

  10. Researches about selecting resistant melon types to fusarium oxyporum f. sp.melonis race 1,2 by using tissue culture and mutation techniques

    International Nuclear Information System (INIS)

    2009-01-01

    breeding was quickly understood. In vitro techniques for crop improvement first consisted of micropropagation and plant regeneration, and then in vitro methods were also found to be useful for eliminating disease and selecting for resistant cells or explants. In vitro selection using specific chemical compounds and pathogens is another useful aspect of tissue culture. More recently, in vitro techniques were combined to mutation induction for generating genetic variation, including novel disease resistant mutants. Mutation induction can be caused by chemical or physical mutagens that alter the structure of the DNA. Treatment of in vitro tissues with physical or chemical mutagens may increase the frequency of genetic variation considerably. The physical mutagens most commonly used are X-rays, gamma rays and UV light, whereas Ethyl Methane- Sulphonate (EMS) is the chemical mutagen most used in crop improvement. Irradiation treatments may be a suitable choice of mutagen for a number of reasons because the application is fast and, in contrast to chemical mutagens, there is no risk that residues remain in the medium. In this research we are going to determine the resistant cells, which will come from irradiated and non-irradiated explants by using races of F. oxysporum f. sp. melonis culture filtrates in vitro conditions. If we will get successful results by this method we will be able to develop new melon cultivars, which will be resistant to F. oxysporum f. sp. melonis. The present work was performed with in vitro plantlets obtained from seeds of melon cv. Yuva which is an important commercial cultivar in Turkey. According to our observations filtrate and gamma ray treatment have an important effect on tolerant plantlet and callus formation. In this research, we show a method for mass-selection of melon mutants resistant to Fusarium wilt. In vitro selection of resistant cells, which are come from irradiated and non-irradiated explants, is done using culture filtrates of

  11. A Técnica Esportiva como Construção Cultural: implicações para a pedagogia do esporte Sport Technique as a Cultural Construction: Implications to Sport Pedagogy La técnica deportiva como construcción cultural: implicaciones hacia la pedagogía del deporte

    Directory of Open Access Journals (Sweden)

    2008-03-01

    Full Text Available

    O texto discute a técnica esportiva a partir de alguns referenciais das ciências sociais e humanas, fazendo um contraponto ao conceito tradicional de técnica utilizado pela área de Educação Física e Esporte, cuja base teórica é composta primordialmente por contribuições das ciências da natureza. Considerando a técnica como o conjunto de modos de fazer e a tática como as razões do fazer, o texto trata das implicações para a pedagogia do esporte, propondo o ensino da cultura esportiva de forma mais democrática, singular e autoral, respeitando as especificidades culturais e as constantes ressignificações do esporte.

    PALAVRAS-CHAVE: esporte - cultura esportiva - pedagogia do esporte - técnica esportiva

    This text discusses technique in sport based on references from the social and human sciences and it questions the traditional concept of technique used in the field of physical education and sports, which is taken primarily from contributions of the natural sciences. By considering technique as a set of procedures to do something, and tactics as the reasons for doing something, the text discusses its possible implications for sport pedagogy and argues for a teaching of sport culture that is more democratic, singular, and authoral, with a respect for cultural specificities and for the constant resignifications of sports.


    Keywords: sport – sport culture – sport pedagogy – sport technique.

    El texto discute la técnica deportiva partiendo de algunos referenciales de las ciencias sociales y humanas, y hace un contrapunto al concepto tradicional de técnica utilizado por el área de Educación Física y Deporte, cuya base teórica se compone primordialmente por contribuciones de las ciencias naturales. Considerando la técnica como el conjunto de maneras de hacer, y la táctica como las razones de hacer, el texto trata de las implicaciones para la pedagogía deportiva, y propone la ense

  12. Transformation and mass hyperplasia technique of the garden plant (lily) by radiation and so forth. Mass hyperplasia of the lily using tissue culture

    International Nuclear Information System (INIS)

    Shigematsu, Koji; Hamada, Yutaka

    1997-01-01

    For an aim of more uniform child bulb production and good quality kind conservation using tissue culture of the lily, some hyperplasia from organs over ground of the lily were tried. In particular, optimum culture media with higher hyperplasia rate of the child bulb, redifferentiation due to difference among kinds of the lilies, and difference of hyperplasia of the child bulbs were investigated. As a result, it was found that pollution due to various germs attached to used materials often occurs, that efficiency obtainable for initial child bulb by redifferentiation from the organs was low at 20%, and that pollution due to various germs was often found at 25degC of cultivation temperature, which was inferior to that at 20degC. And, when conducting mass hyperplasia of the lily using tissue culture, an optimum culture medium of formation and hyperplasia of child bulb could be obtained for its each kind. As a result of conducting some investigations on configuration of the lily nourished from its child bulb and flowered by the tissue culture, it was also found that cultured bulb had the same character as its parent bulb had. (G.K.)

  13. Direct identification of microorganisms from positive blood cultures using the lysis-filtration technique and matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS): a multicentre study.

    Science.gov (United States)

    Farina, Claudio; Arena, Fabio; Casprini, Patrizia; Cichero, Paola; Clementi, Massimo; Cosentino, Marina; Degl'Innocenti, Roberto; Giani, Tommaso; Luzzaro, Francesco; Mattei, Romano; Mauri, Carola; Nardone, Maria; Rossolini, Gian Maria; Serna Ortega, Paula Andrea; Vailati, Francesca

    2015-04-01

    Microbial identification from blood cultures is essential to institute optimal antibiotic therapy and improve survival possibilities. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been successfully applied to identify bacteria and yeasts from positive blood cultures broths. The aim of this multicentre study was to evaluate the reliability of the lysis-filtration technique associated with MALDI-TOF MS to directly identify microorganisms from 765 positive blood cultures collected in six Italian hospitals. Overall, 675/765 (78.1%) blood isolates were correctly identified at the species level, with significant differences between Gram-negative and Gram-positive bacteria (92.6%, and 69.8%, respectively). Some difficulties arise in identifying Streptococcus pneumoniae, Staphylococcus aureus, yeasts and anaerobes. The lysis-filtration protocol is a suitable procedure in terms of performance in identifying microorganisms, but it is quite expensive and technically time-consuming since the time of filtration is not regular for all the samples. The application of the MALDI-TOF MS technique to the direct microbial identification from positive blood cultures is a very promising approach, even if more experience must be gained to minimize errors and costs.

  14. Culture de suspensions cellulaires embryogéniques et régénération en plantules par embryogenèse somatique chez le bananier et le bananier plantain Musa spp

    Directory of Open Access Journals (Sweden)

    Dhed'a, D.

    1992-01-01

    Full Text Available Embryogenie cell suspension and plant regeneration through somatic embryogenesis in bananas and plantains Musa spp. Embryogenie cell suspensions have been initiated using explants from meristematic shoot-tips (scalps. The culture medium has been a modified Murashige and Skoog medium supplemented, according to the steps of culture, with 5fiM 2, 4-D, 1-10//M BAPorzeatin. The suspensions obtained for 5 banana varieties have regenerated plants through somatic embryogenesis. Embryogenie cell suspensions have proved to be the material of choice for cryopreservation, protoplast isolation and culture and for genetic manipulation of Musa for resistance to diseases.

  15. Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. [Annual report], May 16, 1993--January 29, 1994

    Energy Technology Data Exchange (ETDEWEB)

    Steponkus, P.L.

    1994-06-01

    Our aim is to provide a mechanistic understanding of the cellular and molecular aspects of freezing injury and cold acclimation from a perspective of the structural and functional integrity of the plasma membrane-the primary site of freezing injury in winter cereals. We established that destabilization of the plasma membrane of winter rye, the most freezing-tolerant winter cereal, can result from several different lesions: expansion induced lysis, lamellar-to-hexagonal II phase transitions, and the fracture-jump lesion. The occurrence and incidence of these various lesions, depends on the freeze/thaw protocol and the stage of cold acclimation. In non-acclimated leaves and protoplasts, expansion-induced lysis is the predominant lesion at temperatures between {minus}2 and {minus}5{degree}C, whereas freeze-induced formation of the H{sub II} phase is the predominant lesion at temperatures below {minus}10{degree}C. We investigated whether the difference in freezing tolerance and the threshold temperatures at which the lesions occur in rye and oat are a consequence of differences in the lipid composition of the plasma membrane. There are substantial differences between rye and oat cell membranes both before and after cold acclimation. The plasma membrane of oat contains greater proportions of acylated sterylglucosides and cerebrosides than that of rye, and there is little change in these two lipid classes during cold acclimation. The lyotropic phase behavior of lipid mixtures that resemble the plasma membrane of rye and oat was studied. The differences in lipid composition of rye and oat are of mechanistic significance because of their influence on the hydration characteristics of the plasma membrane, the propensity for dehydration-induced lipid-lipid demixing, and the intrinsic curvature of the lipid monolayers. These studies suggest that strategies for improving the freezing tolerance of winter cereals should include approaches to modify membrane lipid composition.

  16. Production de minitubercules de pomme de terre par hydroponie: évaluation d'un système combinant les techniques NFT et Gravel Culture pour deux types de solutions nutritives

    Directory of Open Access Journals (Sweden)

    Rolot J.L.

    2002-01-01

    Full Text Available Potato minituber production through hydropony: assessment of a system combining the NFT and Gravel Culture techniques for two types of nutrient solutions. The potato minituber production is the classical intermediate stage enabling field use of potato materials with an in vitro origin. This production of minitubers may be achieved through various techniques. Most often however they are based upon bedding vitroplantlets or vitroplantlets cuttings in an organic substratum which is disinfected or not. The soilless culture of plants stemming from vitrotubers to produce minitubers with a superior health quality was tested within a hydroponic system. Two types of nutrient solutions were compared: a nitrogen rich one (NPK, mg/l, 180-40-300 and a phosphorus rich one (NPK, mg/l, 60-150-300. In order to initiate the cultures, presprouting microtubers of several varieties (Bintje, Kennebec, Spunta, Saturna, Desiree and Gasore were used. Multiplication rates ranged between 13.2 (Saturna and 3.8 (Kennebec minitubers with a grade higher than 10 mm (more than 1.5 g. As the selected density of the plants was 59 plants per square metre, the yield per square metre varied from 224 to 779 minitubers with a grade higher than 10 mm. The obtained number of minitubers depended especially on the variety. The phosphorus enrichment of the nutrient solution induced an increased number of minitubers produced with a grade higher than 15 mm (more than 5 g. The health state of the produced tubers was excellent.

  17. Detection of microorganisms in culture medium through the neutron radiographic technique; Deteccao de microorganismos em meios de cultura pela tecnica de neutrongrafia

    Energy Technology Data Exchange (ETDEWEB)

    Wacha, Reinaldo

    1999-05-01

    The study aims to obtain a more effective and faster method for the detection of bacteria in several culture media, such as potable water and blood. After the process growth in the culture medium, separation and suspension in buffer solution based in boron, the bacteria are deposited in track detectors that are submitted to thermal neutron beams (neutron flux: 2,2 x 10{sup 5} n.cm{sup -2}.s{sup -1}), resulting from the channel J-9 of the Argonauta research reactor, from the Nuclear Engineering Institute, IEN/CNEN. The latent tracks arisen from the alpha particles proceeding from the reaction B ({eta}, {alpha}) Li and, after having been revealed, are analyzed by an optical microscope that allows to detect the existence of the bacteria. Afterwards, they were analyzed in a nanoscope which helps the identification of the tracks of the alpha particles. (author)

  18. Beyond Blood Culture and Gram Stain Analysis: A Review of Molecular Techniques for the Early Detection of Bacteremia in Surgical Patients.

    Science.gov (United States)

    Scerbo, Michelle H; Kaplan, Heidi B; Dua, Anahita; Litwin, Douglas B; Ambrose, Catherine G; Moore, Laura J; Murray, Col Clinton K; Wade, Charles E; Holcomb, John B

    2016-06-01

    Sepsis from bacteremia occurs in 250,000 cases annually in the United States, has a mortality rate as high as 60%, and is associated with a poorer prognosis than localized infection. Because of these high figures, empiric antibiotic administration for patients with systemic inflammatory response syndrome (SIRS) and suspected infection is the second most common indication for antibiotic administration in intensive care units (ICU)s. However, overuse of empiric antibiotics contributes to the development of opportunistic infections, antibiotic resistance, and the increase in multi-drug-resistant bacterial strains. The current method of diagnosing and ruling out bacteremia is via blood culture (BC) and Gram stain (GS) analysis. Conventional and molecular methods for diagnosing bacteremia were reviewed and compared. The clinical implications, use, and current clinical trials of polymerase chain reaction (PCR)-based methods to detect bacterial pathogens in the blood stream were detailed. BC/GS has several disadvantages. These include: some bacteria do not grow in culture media; others do not GS appropriately; and cultures can require up to 5 d to guide or discontinue antibiotic treatment. PCR-based methods can be potentially applied to detect rapidly, accurately, and directly microbes in human blood samples. Compared with the conventional BC/GS, particular advantages to molecular methods (specifically, PCR-based methods) include faster results, leading to possible improved antibiotic stewardship when bacteremia is not present.

  19. The role of activated charcoal in plant tissue culture.

    Science.gov (United States)

    Thomas, T Dennis

    2008-01-01

    Activated charcoal has a very fine network of pores with large inner surface area on which many substances can be adsorbed. Activated charcoal is often used in tissue culture to improve cell growth and development. It plays a critical role in micropropagation, orchid seed germination, somatic embryogenesis, anther culture, synthetic seed production, protoplast culture, rooting, stem elongation, bulb formation etc. The promotary effects of AC on morphogenesis may be mainly due to its irreversible adsorption of inhibitory compounds in the culture medium and substancially decreasing the toxic metabolites, phenolic exudation and brown exudate accumulation. In addition to this activated charcoal is involved in a number of stimulatory and inhibitory activities including the release of substances naturally present in AC which promote growth, alteration and darkening of culture media, and adsorption of vitamins, metal ions and plant growth regulators, including abscisic acid and gaseous ethylene. The effect of AC on growth regulator uptake is still unclear but some workers believe that AC may gradually release certain adsorbed products, such as nutrients and growth regulators which become available to plants. This review focuses on the various roles of activated charcoal in plant tissue culture and the recent developments in this area.

  20. Produção de protoplastos e lise da parede celular de leveduras utilizando β-1,3 glucanase Protoplasts production and yeast cell wall lysis using β-1,3 glucanase

    Directory of Open Access Journals (Sweden)

    Luciana Francisco Fleuri

    2010-06-01

    Full Text Available O presente trabalho visou a aplicação da β-1,3 glucanase lítica, obtida do microrganismo Cellulosimicrobium cellulans 191, na produção de protoplastos e na lise da parede celular de leveduras. A preparação bruta da enzima foi capaz de lisar as leveduras Kluyveromyces lodderi, Saccharomyces cerevisiae (Fleischmann e Itaiquara, S. cerevisiae KL-88, S. diastaticus NCYC 713, S. cerevisiae NCYC 1001, Candida glabrata NCYC 388, Kluyveromyces marxianus NCYC 587 e Hansenula mrakii NCYC 500. A β-1,3 glucanase purificada foi capaz de lisar as leveduras Saccharomyces cerevisiae KL-88, Saccharomyces capensis, Debaromyces vanriji, Pachysolen tannophillus, Kluyveromyces drosophilarum, Candida glabrata, Hansenula mrakii e Pichia membranaefaciens e formar protoplastos de Saccharomyces cerevisiae KL-88.The aim of this work was the application of lytic β-1,3 glucanase obtained from Cellulosimicrobium cellulans strain 191 in the production of protoplasts and lysis of yeast cell walls. The crude extract demonstrated lysis activity against the yeasts Kluyveromyces lodderi, Saccharomyces cerevisiae (Fleischmann and Itaiquara, S. cerevisiae KL-88, S. diastaticus NCYC 713, S. cerevisiae NCYC 1001, Candida glabrata NCYC 388, Kluyveromyces marxianus NCYC 587, and Hansenula mrakii NCYC 500. The purified β-1,3 glucanase demonstrated lysis activity against the yeasts Saccharomyces cerevisiae KL-88, Saccharomyces capensis, Debaromyces vanriji, Pachysolen tannophillus, Kluyveromyces drosophilarum, Candida glabrata, Hansenula mrakii, and Pichia membranaefaciens, and it was able to produce Saccharomyces cerevisiae KL-88 protoplasts.

  1. Drug uptake (DAPI) of trypanosomes (T. brucei) and antitrypanosomal activity in vitro, in culture and in vivo studied by microscope fluorometry, chromatogram spectrophotometry and radiotracer techniques

    International Nuclear Information System (INIS)

    Kratzer, R.D.

    1982-01-01

    The present study had the following objectives: 1) Investigation of the specific binding and location of the diamidine DAPI within trypanosomes by fluorescence microscopy. 2) Development and standardization of a microscope fluorometry technique for measuring DAPI uptake of single trypanosomes. 3) Determination of the effect of incubation media, exposure time, and drug concentration on DAPI uptake of single trypanosomes. 4) Development of a technique applicable for quantitative fluorescence chemical analysis of DAPI uptake of trypanosomes. 5) Determination of drug uptake of trypanosomes using 14 C labelled DAPI. 6) Comparison of the values obtained by the three methods. (orig./MG)

  2. Personal Space Across Cultures

    DEFF Research Database (Denmark)

    Høgh-Olesen, Henrik

    2017-01-01

    —constitutes one of the main areas of interest, dividing cultures into either contact or low-contact cultures. Examples of proxetics—human spatial behavior studied in terms of universal spacing patterns and cultural similarities—are presented. Finally, future directions for the study of PS in the digital age...... functions. The integrity zone has no fixed size but varies according to variables such as age, gender, personality, relation, and culture. The key theoretical traditions and models are presented and the field's methodological techniques and measurements are discussed. Proxemics—cultural differences...

  3. Corpora and Cultural Cognition

    DEFF Research Database (Denmark)

    Jensen, Kim Ebensgaard

    2017-01-01

    Cultural cognition is, to a great extent, transmitted through language and, consequently, reflected and replicated in language use. Cultural cognition may be instantiated in various patterns of language use, such as the discursive behavior of constructions. Very often, such instantiations can be ...... is addressed. In the third part of the chapter, three case studies are presented – one from Danish and two from English – to illustrate the analysis of cultural conceptualization via corpus-linguistic techniques....

  4. Cultural neurolinguistics.

    Science.gov (United States)

    Chen, Chuansheng; Xue, Gui; Mei, Leilei; Chen, Chunhui; Dong, Qi

    2009-01-01

    As the only species that evolved to possess a language faculty, humans have been surprisingly generative in creating a diverse array of language systems. These systems vary in phonology, morphology, syntax, and written forms. Before the advent of modern brain-imaging techniques, little was known about how differences across languages are reflected in the brain. This chapter aims to provide an overview of an emerging area of research - cultural neurolinguistics - that examines systematic cross-cultural/crosslinguistic variations in the neural networks of languages. We first briefly describe general brain networks for written and spoken languages. We then discuss language-specific brain regions by highlighting differences in neural bases of different scripts (logographic vs. alphabetic scripts), orthographies (transparent vs. nontransparent orthographies), and tonality (tonal vs. atonal languages). We also discuss neural basis of second language and the role of native language experience in second-language acquisition. In the last section, we outline a general model that integrates culture and neural bases of language and discuss future directions of research in this area.

  5. Development, Characterization and Cell Cultural Response of 3D Biocompatible Micro-Patterned Poly-ε-Caprolactone Scaffolds Designed and Fabricated Integrating Lithography and Micromolding Fabrication Techniques

    KAUST Repository

    Limongi, Tania; Miele, Ermanno; Shalabaeva, Victoria; Rocca, Rosanna La; Schipani, Rossana; Malara, Natalia; Angelis, Francesco de; Giugni, Andrea; Di Fabrizio, Enzo M.

    2014-01-01

    Scaffold design and fabrication are very important subjects for biomaterial, tissue engineering and regenerative medicine research playing a unique role in tissue regeneration and repair. Among synthetic biomaterials Poly-ε- Caprolactone (PCL) is very attractive bioresorbable polyester due to its high permeability, biodegradability and capacity to be blended with other biopolymers. Thanks to its ability to naturally degrade in tissues, PCL has a great potential as a new material for implantable biomedical micro devices. This work focuses on the establishment of a micro fabrication process, by integrating lithography and micromolding fabrication techniques, for the realization of 3D microstructure PCL devices. Scaffold surface exhibits a combination in the patterned length scale; cylindrical pillars of 10 μm height and 10 μm diameter are arranged in a hexagonal lattice with periodicity of 30 μm and their sidewalls are nano-sculptured, with a regular pattern of grooves leading to a spatial modulation in the z direction. In order to demonstrate that these biocompatible pillared PCL substrates are suitable for a proper cell growth, NIH/3T3 mouse embryonic fibroblasts were seeded on them and cells key adhesion parameters were evaluated. Scanning Electron Microscopy and immunofluorescence analysis were carried out to check cell survival, proliferation and adhesion; cells growing on the PCL substrates appeared healthy and formed a well-developed network in close contact with the micro and nano features of the pillared surface. Those 3D scaffolds could be a promising solution for a wide range of applications within tissue engineering and regenerative medicine applications.

  6. Development, Characterization and Cell Cultural Response of 3D Biocompatible Micro-Patterned Poly-ε-Caprolactone Scaffolds Designed and Fabricated Integrating Lithography and Micromolding Fabrication Techniques

    KAUST Repository

    Limongi, Tania

    2014-12-12

    Scaffold design and fabrication are very important subjects for biomaterial, tissue engineering and regenerative medicine research playing a unique role in tissue regeneration and repair. Among synthetic biomaterials Poly-ε- Caprolactone (PCL) is very attractive bioresorbable polyester due to its high permeability, biodegradability and capacity to be blended with other biopolymers. Thanks to its ability to naturally degrade in tissues, PCL has a great potential as a new material for implantable biomedical micro devices. This work focuses on the establishment of a micro fabrication process, by integrating lithography and micromolding fabrication techniques, for the realization of 3D microstructure PCL devices. Scaffold surface exhibits a combination in the patterned length scale; cylindrical pillars of 10 μm height and 10 μm diameter are arranged in a hexagonal lattice with periodicity of 30 μm and their sidewalls are nano-sculptured, with a regular pattern of grooves leading to a spatial modulation in the z direction. In order to demonstrate that these biocompatible pillared PCL substrates are suitable for a proper cell growth, NIH/3T3 mouse embryonic fibroblasts were seeded on them and cells key adhesion parameters were evaluated. Scanning Electron Microscopy and immunofluorescence analysis were carried out to check cell survival, proliferation and adhesion; cells growing on the PCL substrates appeared healthy and formed a well-developed network in close contact with the micro and nano features of the pillared surface. Those 3D scaffolds could be a promising solution for a wide range of applications within tissue engineering and regenerative medicine applications.

  7. Securing a better future for all: Nuclear techniques for global development and environmental protection. NA factsheet on nuclear sciences and applications collaborating centres: Establishing a culture of cooperation

    International Nuclear Information System (INIS)

    2012-01-01

    As a large international organization with a variety of programmes, initiatives and laboratories, one of the mandates of the IAEA is to promote the peaceful use of nuclear techniques worldwide while also collaborating with other institutions in its Member States in order to implement part of its approved activities. In this regard, the IAEA designates selected institutions as official IAEA collaborating centres (IAEA-CCs). Recognizing the need to preserve and transfer nuclear knowledge, the IAEA-CCs are dedicated to furthering the research, development and training in peaceful applications of nuclear science and technology. By working alongside various nuclear related institutions from around the world, the IAEA contributes to reaching important targets laid out by the United Nations Millennium Development Goals. The IAEA-CCs are, in general, scientific institutions such as laboratories, universities, research facilities, etc., that receive public recognition by the IAEA and have been designated to collaborate with the IAEA in a variety of fields, such as food safety, environmental protection, water resources and human health. In line with the objectives of the IAEA, IAEA-CCs are expected to further the research, development and training in the peaceful applications of nuclear science and technology. Through these research and academic institutions, Member States can assist the IAEA with their own original research, development and training in nuclear technologies. As a cooperative mechanism, the IAEA-CC is also efficient, as it encourages centres to share resources, knowledge and expertise. One of the goals of the IAEA-CC is also to help developing Member States expand their capabilities in these areas and thus improve the quality of life of their citizens. So far, the IAEA-CC has led to socioeconomic benefits in many parts of the world, from Asia to Latin America.

  8. Safety culture in nuclear industry

    International Nuclear Information System (INIS)

    Sundararajan, A.R.

    1998-01-01

    This paper after defining the term safety culture outlines the requirements at various levels of the plant management to ensure that safety culture pervades all activities related to the plant. Techniques are also indicated which can be used to assess the effectiveness of safety culture

  9. Tools to quantify safety culture

    International Nuclear Information System (INIS)

    Avella, B.

    2011-01-01

    This paper reviews the notion of safety culture and then describes some of the tools that can be used to assess it. Required characteristics to obtain reliable tools and techniques are provided, along with a short summary of the most common and important tools and techniques used to assess safety culture at the nuclear field is described. At the end of this paper, the reader will better understand the importance of the safety culture of the organization and will have requirements to help him in choosing reliable tools and techniques. Further, there will be recommendations on how best to follow-up after an assessment of safety culture. (author)

  10. Urine culture

    Science.gov (United States)

    Culture and sensitivity - urine ... when urinating. You also may have a urine culture after you have been treated for an infection. ... when bacteria or yeast are found in the culture. This likely means that you have a urinary ...

  11. Safeguards Culture

    Energy Technology Data Exchange (ETDEWEB)

    Frazar, Sarah L.; Mladineo, Stephen V.

    2012-07-01

    The concepts of nuclear safety and security culture are well established; however, a common understanding of safeguards culture is not internationally recognized. Supported by the National Nuclear Security Administration, the authors prepared this report, an analysis of the concept of safeguards culture, and gauged its value to the safeguards community. The authors explored distinctions between safeguards culture, safeguards compliance, and safeguards performance, and evaluated synergies and differences between safeguards culture and safety/security culture. The report concludes with suggested next steps.

  12. Organizational culture

    OpenAIRE

    Schein, Edgar H.

    1988-01-01

    Cultural orientations of an organization can be its greatest strength, providing the basis for problem solving, cooperation, and communication. Culture, however, can also inhibit needed changes. Cultural changes typically happen slowly – but without cultural change, many other organizational changes are doomed to fail. The dominant culture of an organization is a major contributor to its success. But, of course, no organizational culture is purely one type or another. And the existence of sec...

  13. Experimental techniques; Techniques experimentales

    Energy Technology Data Exchange (ETDEWEB)

    Roussel-Chomaz, P. [GANIL CNRS/IN2P3, CEA/DSM, 14 - Caen (France)

    2007-07-01

    This lecture presents the experimental techniques, developed in the last 10 or 15 years, in order to perform a new class of experiments with exotic nuclei, where the reactions induced by these nuclei allow to get information on their structure. A brief review of the secondary beams production methods will be given, with some examples of facilities in operation or under project. The important developments performed recently on cryogenic targets will be presented. The different detection systems will be reviewed, both the beam detectors before the targets, and the many kind of detectors necessary to detect all outgoing particles after the reaction: magnetic spectrometer for the heavy fragment, detection systems for the target recoil nucleus, {gamma} detectors. Finally, several typical examples of experiments will be detailed, in order to illustrate the use of each detector either alone, or in coincidence with others. (author)

  14. Recent developments in the techniques of IMAG processing and diagnostic imaging for the cultural heritage; Recenti sviluppi nelle metodologie di elaborazione di immagini e di imaging diagnostico per i beni culturali

    Energy Technology Data Exchange (ETDEWEB)

    Maino, G. [ENEA, Divisione Fisica Applicata, Centro Ricerche E. Clementel, Bologna (Italy); Bonifazzi, C. [Ferrara Univ., Ferrara (Italy). Dipt. di Scienze Biomediche; Sabia, E. [ENEA, Tecnologie Ingegneria e Servizi Ambientali, Centro Ricerche Portici, Naples (Italy)

    2000-07-01

    In this report, the contributions presented to the mini symposium on Recent developments in the techniques of image processing and diagnostic imaging for the cultural heritage are collected. The symposium has been organized within the framework of the V National Congress of SIMAI Italian Society for Applied and Industrial Mathematics (SIMAI 2000), held in Ischia from June 5 to 9, 2000, and shows the main scientific results obtained relevant to the research activities carried out in the frame of sub project 3 of research project GIANO (Advanced Graphics for the National cultural heritage and Employment), jointly funded by MURST (Ministry of University, Scientific and Technological Research) EC (law 488/92). The GIANO-ENEA project contributed, together with CRS4, GNIM-IndAM, Ministero degli Affari Esteri and ST-Microelectronics, to the organization of SIMAI 2000. [Italian] In questo rapporto sono raccolti i contributi presentati al minisimposio su Recenti sviluppi nelle metodologie di elaborazione di immagini e di imaging diagnostico per i beni culturali, organizzato nell'ambito del 5. Congresso Nazionale della SIMAI Societa' Italiana di Matematica Applicata e Industriale (SIMAI 2000), svoltosi a Ischia dal 5 al 9 giugno 2000. Il simposio illustra i principali risultati scientifici ottenuti nel quadro delle attivita' di ricerca del sottoprogetto 3 del piano nazionale GIANO (Grafica Innovativa per il patrimonio Artistico Nazionale e per l'Occupazione giovanile), finanziato dal MURST e dall'UE (legge 488/92). Il progetto GIANO-ENEA ha contribuito, insieme con CRS4, GNIM-IndAM, Ministero degli Affari Esteri e ST-Microelectronics, alla realizzazione di SIMAI 2000.

  15. Hyaluronan synthesis in cultured tobacco cells (BY-2) expressing a chlorovirus enzyme: cytological studies.

    Science.gov (United States)

    Rakkhumkaew, Numfon; Shibatani, Shigeo; Kawasaki, Takeru; Fujie, Makoto; Yamada, Takashi

    2013-04-01

    Extraction of hyaluronan from animals or microbial fermentation has risks including contamination with pathogens and microbial toxins. In this work, tobacco cultured-cells (BY-2) were successfully transformed with a chloroviral hyaluronan synthase (cvHAS) gene to produce hyaluronan. Cytological studies revealed accumulation of HA on the cells, and also in subcellular fractions (protoplasts, miniplasts, vacuoplasts, and vacuoles). Transgenic BY-2 cells harboring a vSPO-cvHAS construct containing the vacuolar targeting signal of sporamin connected to the N-terminus of cvHAS accumulated significant amounts of HA in vacuoles. These results suggested that cvHAS successfully functions on the vacuolar membrane and synthesizes/transports HA into vacuoles. Efficient synthesis of HA using this system provides a new method for practical production of HA. Copyright © 2012 Wiley Periodicals, Inc.

  16. Cultural entrepreneurship

    NARCIS (Netherlands)

    A. Klamer (Arjo)

    2011-01-01

    textabstractCultural entrepreneurship is a new character in the cultural sector. This paper characterizes the cultural entrepreneur paying homage to the hermeneutic approach of Don Lavoie and others. The challenge is to render the "cultural" meaningful. An invention is the highlighting of the

  17. Industrial cultures

    DEFF Research Database (Denmark)

    Rasmussen, Lauge Baungaard

    1996-01-01

    The chapter deals with different paradigms andtheories of cultural development. The problem toexplain change and methods to analyse developmentin different cultures are presented and discussed.......The chapter deals with different paradigms andtheories of cultural development. The problem toexplain change and methods to analyse developmentin different cultures are presented and discussed....

  18. OBTENÇÃO DE PLANTAS DE LIMÃO CRAVO (Citrus limonia Osbeck E TANGERINA CLEÓPATRA (Citrus reshni Hort. A PARTIR DO CULTIVO DE PROTOPLASTOS DE SUSPENSÃO CELULAR PLANT REGENERATION OF 'RANGPUR' LIME (Citrus limonia Osbeck AND 'CLEÓPATRA' MANDARIN (Citrus reshni Hort. THROUGH PROTOPLASTS OF CELL SUSPENSION

    Directory of Open Access Journals (Sweden)

    Rodrigo Rocha Latado

    1999-01-01

    Full Text Available Este trabalho descreve uma metodologia para a regeneração de plantas de tangerina 'Cleópatra' e limão 'Cravo', a partir do cultivo de protoplastos de suspensão celular. Para tal, calos nucelares foram induzidos em meio contendo BAP e cultivados em meio sem reguladores de crescimento. Protoplastos foram isolados de suspensões celulares e cultivados em gotas de agarose, com densidade de 2 X 105 protoplastos.ml-1. O meio MT, contendo ácido giberélico e água de coco, foi eficiente na germinação de embriões somáticos. Os métodos de aclimatação de plantas testados apresentaram baixa eficiência. Como resultado final, 17 plantas adaptadas de tangerina e 8 de limão foram obtidas.The present research describes the regeneration of 'Cleópatra' mandarin and 'Rangpur' lime plants from cell suspension protoplasts. Nucelar calli were induced on a medium containing BAP and maintained on growth regulator free medium. Protoplasts were isolated from embryogenic suspension and plated at a concentration of 2 X 105 protoplasts.ml-1, on agarose droplets. The MT medium with gibberellic acid and coconut water was efficient to stimulate somatic embryo conversion. Rooted plants acclimation had low efficiency. Seventeen mandarin plants and eight lime plants were obtained.

  19. Morphological differentiation of Streptomyces viridochromogenes E-219 on solid culture

    International Nuclear Information System (INIS)

    Liang Xinle; Zhu Jing; Jin Yingyan

    2012-01-01

    The Streptomyces viridochromogenes E-219 was derived from Streptomyces viridochromogenes CGMCC4.1119 treated with 60 Co γ-rays irradiation and protoplast fusion. With the help of fluorescent probes, fluorescence microscope and electron microscopy, the morphology and development of E-219 on solid surface culture were investigated in this study. The effect of agarslant culture time on the production of Avilamycin was also studied to provide theoretical basis for industrial fermentation of selecting the appropriate seed to culture on the agarslant culture medium. The results implied that the development of colonies of Streptomyces viridochromogenes accompanied the intermittent hyhae apoptosis, and the production of spores was from the active mycelium. The colonial morphology of strain E-219 was significantly different from the original strain CGMCC4h1119. There were variegated hyphae formation in the stage of spore germination and initial hyphae development (10 h) with the live and dead segments alternated in a highly regular fashion within the same hypha. After the early single colony formation, the third phase was followed by profuse growth of the live segments derived from the variegated hypha, then the second apoptosis of the mycelia (48 h) was occurred with another quick growth, and sporulation was occurred at 96 h. Strain CGMCC4.1119 had spiral sporotrichial and round conidiophores with spike, whereas strain E-219 had linear sporotrichial, smooth and dylindrical conidiophore. The results of shake flask experiments indicated that the spores of E-219 had that highest activity when cultured on agarslant culture medium and incubated for 106 h with the production of avilamycin up to 1200 mg/L. (authors)

  20. ORGANIZATIONAL CULTURE AND MANAGEMENT CULTURE

    OpenAIRE

    Tudor Hobeanu; Loredana Vacarescu Hobeanu

    2010-01-01

    Communication reveals the importance of organizational culture and management culture supported by the remarkable results in economic and social level of organization. Their functions are presented and specific ways of expression levels of organizational culture and ways of adapting to the requirements of the organization's management culture.

  1. Cultural commons and cultural evolution

    OpenAIRE

    Bravo, Giangiacomo

    2010-01-01

    Culture evolves following a process that is akin to biological evolution, although with some significant differences. At the same time culture has often a collective good value for human groups. This paper studies culture in an evolutionary perspective, with a focus on the implications of group definition for the coexistence of different cultures. A model of cultural evolution is presented where agents interacts in an artificial environment. The belonging to a specific memetic group is a majo...

  2. The Cultural Content of Business Spanish Texts.

    Science.gov (United States)

    Grosse, Christine Uber; Uber, David

    A study examined eight business Spanish textbooks for cultural content by looking at commonly appearing cultural topics and themes, presentation of cultural information, activities and techniques used to promote cultural understanding, and incorporation of authentic materials. The texts were evenly divided among beginning, intermediate, and…

  3. Ciência, técnica e cultura: relações entre risco e práticas de saúde Science, technique, and culture: relations between risk and health practices

    Directory of Open Access Journals (Sweden)

    Dina Czeresnia

    2004-04-01

    Full Text Available Este artigo tem como objetivo discutir conseqüências culturais dos discursos e práticas voltados à capacitação dos sujeitos para a escolha racional e informada de riscos, calculados com base no conhecimento científico. O conceito de risco epidemiológico é um dos elementos centrais deste processo, especialmente no contexto das práticas de saúde. Inicialmente é apresentada uma breve caracterização do conceito de risco epidemiológico, ressaltando que, como modelo abstrato, reduz a complexidade dos fenômenos que estuda. A apreensão da realidade mediante essa abstração gera valores e significados. A reflexão de Canguilhem sobre as relações entre ciência, técnica e vida é retomada com a perspectiva de aprofundar a compreensão das conseqüências culturais produzidas, que contribuem para a transformação das concepções clássicas de individualidade, autonomia e sociabilidade. Temas vitais cruciais, como individualidade, alteridade, relação com a morte, estão presentes (ocultos nas questões que envolvem a nuclearidade do risco no mundo atual.This article discusses the cultural consequences of discourses and practices aimed at training subjects for a rational, informed choice in relation to risks, calculated on the basis of scientific knowledge. The epidemiological risk concept is a central element in this process, especially in the context of health practices. The article begins by briefly characterizing the epidemiological risk concept, emphasizing that as an abstract model, it reduces the complexity of the phenomena it studies. Grasping reality through this abstraction generates values and meanings. Canguilhem's reflection on the relations between science, technique, and life is further discussed from the perspective of deepening an understanding of the cultural consequences of this process, contributing to the transformation of classical concepts of individuality, autonomy, and sociability. Such vital themes as

  4. Culturing Protozoa.

    Science.gov (United States)

    Stevenson, Paul

    1980-01-01

    Compares various nutrient media, growth conditions, and stock solutions used in culturing protozoa. A hay infusion in Chalkey's solution maintained at a stable temperature is recommended for producing the most dense and diverse cultures. (WB)

  5. Citrus asymmetric somatic hybrids produced via fusion of gamma-irradiated and iodoacetamide-treated protoplasts Híbridos somáticos assimétricos de citros produzidos pela fusão de protoplastos irradiados e tratados com iodoacetamida

    Directory of Open Access Journals (Sweden)

    Claudine Maria de Bona

    2009-05-01

    Full Text Available The objective of this study was to produce citrus somatic asymmetric hybrids by fusing gamma-irradiated protoplasts with iodoacetamide-treated protoplasts. Protoplasts were isolated from embryogenic suspension cells of grapefruit (Citrus paradisi Macfad. cultivars Ruby Red and Flame, sweet oranges (C. sinensis Osbeck 'Itaboraí', 'Natal', Valencia', and 'Succari', from 'Satsuma' (C. unshiu Marcow. and 'Changsha' mandarin (C. reticulata Blanco and 'Murcott' tangor (C. reticulata x C. sinensis. Donor protoplasts were exposed to gamma rays and receptor protoplasts were treated with 3 mmol L-1 iodoacetamide (IOA, and then they were fused for asymmetric hybridization. Asymmetric embryos were germinated, and the resulting shoots were either grafted onto sour orange, rough lemon or 'Swingle' (C. paradisi x Poncirus trifoliata x 'Sunki' mandarin rootstock seedlings, or rooted after dipping their bases in indol-butyric acid (IBA solution. The products were later acclimatized to greenhouse conditions. Ploidy was analyzed by flow cytometry, and hybridity was confirmed by amplified fragment length polymorphism (AFLP analysis of plantlet DNAsamples. The best treatment was the donor-recipient fusion combination of 80 Gy-irradiated 'Ruby Red' protoplasts with 20 min IOA-treated 'Succari' protoplasts. Tetraploid and aneuploid plants were produced. Rooting recalcitrance was solved by dipping shoots' stems in 3,000 mg L-1 IBA solution for 10 min.O objetivo deste trabalho foi produzir híbridos somáticos assimétricos de citros pela fusão de protoplastos irradiados com raios gama e protoplastos tratados com iodoacetamida. Protoplastos foram isolados de suspensões celulares embriogênicas de pomelo (Citrus paradisi Macfad., cultivares Ruby Red e Flame, de laranja doce (C. sinensis Osbeck 'Itaboraí', 'Natal', Valencia' e 'Succari', de tangerinas 'Satsuma' (C. unshiu Marcow. e 'Changsha' (C. reticulata Blanco e de tangor 'Murcott' (C. reticulata x C. sinensis

  6. Safety culture

    International Nuclear Information System (INIS)

    Keen, L.J.

    2003-01-01

    Safety culture has become a topic of increasing interest for industry and regulators as issues are raised on safety problems around the world. The keys to safety culture are organizational effectiveness, effective communications, organizational learning, and a culture that encourages the identification and resolution of safety issues. The necessity of a strong safety culture places an onus on all of us to continually question whether the safety measures already in place are sufficient, and are being applied. (author)

  7. Beyond Culture.

    Science.gov (United States)

    Barron, Daniel D.

    1993-01-01

    Discusses the lack of literature relating to cultural differences and school library media programs and reviews the book "Beyond Culture" by Edward T. Hall. Highlights include the population/environment crisis, cultural literacy, the use of technology, and Marshall McLuhan's idea of the global village. (LRW)

  8. Bile culture

    Science.gov (United States)

    Culture - bile ... is placed in a special dish called a culture medium to see if bacteria, viruses, or fungi ... Chernecky CC, Berger BJ. Body fluid - anaerobic culture. In: ... . 6th ed. St Louis, MO: Elsevier Saunders; 2013:225-226. Kim AY, ...

  9. Handling Culture

    NARCIS (Netherlands)

    Pieter van Nispen tot Pannerden

    2011-01-01

    The article indicates how companies may prepare for and deal with cultural differences. Because the research base is still rather limited an overall perspective may not be realised. After discussing definitions and concepts of culture, as well as values, cultural differences between states are

  10. Culture of insect tissues

    International Nuclear Information System (INIS)

    Cestari, A.N.; Simoes, L.C.G.

    1978-01-01

    Several aspects are discussed related to the behavior of politenic chromosomes from Rhyncosciara salivary glands kept in culture during different periods of time, without interference of insect hormones. Nucleic acid-and protein synthesis in isolated nuclei and chromosomes are also investigated. Autoradiographic techniques and radioactive precursors for nucleic acids and proteins are used in the research. (M.A.) [pt

  11. Mammalian Cell Culture Simplified.

    Science.gov (United States)

    Moss, Robert; Solomon, Sondra

    1991-01-01

    A tissue culture experiment that does not require elaborate equipment and that can be used to teach sterile technique, the principles of animal cell line maintenance, and the concept of cell growth curves is described. The differences between cancerous and normal cells can be highlighted. The procedure is included. (KR)

  12. 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)

  13. A Theoretical Rationale for Cross-Cultural Family Counseling.

    Science.gov (United States)

    Arciniega, Miguel; Newlon, Betty J.

    1981-01-01

    Proposes seven Adlerian axioms of behavior for the cross-cultural pluralistic counselor working with minority families. Defines cross-cultural family counseling and urges counselors to understand minority cultures and the acculturation process. Discusses counseling techniques. (JAC)

  14. Dismantling techniques

    International Nuclear Information System (INIS)

    Wiese, E.

    1998-01-01

    Most of the dismantling techniques used in a Decontamination and Dismantlement (D and D) project are taken from conventional demolition practices. Some modifications to the techniques are made to limit exposure to the workers or to lessen the spread of contamination to the work area. When working on a D and D project, it is best to keep the dismantling techniques and tools as simple as possible. The workers will be more efficient and safer using techniques that are familiar to them. Prior experience with the technique or use of mock-ups is the best way to keep workers safe and to keep the project on schedule

  15. Dismantling techniques

    Energy Technology Data Exchange (ETDEWEB)

    Wiese, E.

    1998-03-13

    Most of the dismantling techniques used in a Decontamination and Dismantlement (D and D) project are taken from conventional demolition practices. Some modifications to the techniques are made to limit exposure to the workers or to lessen the spread of contamination to the work area. When working on a D and D project, it is best to keep the dismantling techniques and tools as simple as possible. The workers will be more efficient and safer using techniques that are familiar to them. Prior experience with the technique or use of mock-ups is the best way to keep workers safe and to keep the project on schedule.

  16. Spatial Culture

    DEFF Research Database (Denmark)

    Reeh, Henrik

    2012-01-01

    Spatial Culture – A Humanities Perspective Abstract of introductory essay by Henrik Reeh Secured by alliances between socio-political development and cultural practices, a new field of humanistic studies in spatial culture has developed since the 1990s. To focus on links between urban culture...... and modern society is, however, an intellectual practice which has a much longer history. Already in the 1980s, the debate on the modern and the postmodern cited Paris and Los Angeles as spatio-cultural illustrations of these major philosophical concepts. Earlier, in the history of critical studies, the work...... Foucault considered a constitutive feature of 20th-century thinking and one that continues to occupy intellectual and cultural debates in the third millennium. A conceptual framework is, nevertheless, necessary, if the humanities are to adequa-tely address city and space – themes that have long been...

  17. Cultural contrast

    Institute of Scientific and Technical Information of China (English)

    周志

    2016-01-01

    Chinese cultural contains a great number of styles;culture differentiation does not depend on region differentiation.This research would interpret what difference between Hong Kong and Shenzhen.1.Food culture in china Traditional Chinese medicine suggests eating local seasonal fruit and vegetables,as they are most suitable for the body during a particular season.It is also divided food into 3 characteristics:cooling foods,warming foods and balance or

  18. Culture evolves

    OpenAIRE

    Whiten, Andrew; Hinde, Robert A.; Laland, Kevin N.; Stringer, Christopher B.

    2011-01-01

    Culture pervades human lives and has allowed our species to create niches all around the world and its oceans, in ways quite unlike any other primate. Indeed, our cultural nature appears so distinctive that it is often thought to separate humanity from the rest of nature and the Darwinian forces that shape it. A contrary view arises through the recent discoveries of a diverse range of disciplines, here brought together to illustrate the scope of a burgeoning field of cultural evolution and to...

  19. Manuscript Cultures

    DEFF Research Database (Denmark)

    What do Mesoamerica, Greece, Byzantium, Island, Chad, Ethiopia, India, Tibet, China and Japan have in common? Like many other cultures of the world, they share a particular form of cultural heritage: ancient handwritten documents. In 2007, scholars from some20 countries around the world gathered...... at the University of Copenhagen for a workshop on manuscripts to compare notes. This event led to the publication of this volume, which brings together16 articles on philological, cultural, and material aspects of manuscripts in search for a common ground across disciplines and cultures....

  20. Skin or nail culture

    Science.gov (United States)

    Mucosal culture; Culture - skin; Culture - mucosal; Nail culture; Culture - fingernail; Fingernail culture ... There, it is placed in a special dish (culture). It is then watched to see if bacteria, ...

  1. Continuous culture apparatus and methodology

    International Nuclear Information System (INIS)

    Conway, H.L.

    1975-01-01

    At present, we are investigating the sorption of potentially toxic trace elements by phytoplankton under controlled laboratory conditions. Continuous culture techniques were used to study the mechanism of the sorption of the trace elements by unialgal diatom populations and the factors influencing this sorption. Continuous culture methodology has been used extensively to study bacterial kinetics. It is an excellent technique for obtaining a known physiological state of phytoplankton populations. An automated method for the synthesis of continuous culture medium for use in these experiments is described

  2. Cultural probes

    DEFF Research Database (Denmark)

    Madsen, Jacob Østergaard

    The aim of this study was thus to explore cultural probes (Gaver, Boucher et al. 2004), as a possible methodical approach, supporting knowledge production on situated and contextual aspects of occupation.......The aim of this study was thus to explore cultural probes (Gaver, Boucher et al. 2004), as a possible methodical approach, supporting knowledge production on situated and contextual aspects of occupation....

  3. Cultural Communications.

    Science.gov (United States)

    Armas, Jose

    It is too often taken for granted that the communication process with culturally different children takes place as readily as it might with children from Anglo cultures. Most teachers receive training in verbal and formal communication skills; children come to school with nonverbal and informal communication skills. This initially can create…

  4. Connecting Culture

    DEFF Research Database (Denmark)

    Haugegaard, Rikke; Mynster Christensen, Maya

    2017-01-01

    The understanding of cultural dynamics in the area of operations is essential in to the accomplishment of military missions in international operations. The aim of this handbook is to introduce a tool enabling the use of culture as part of the military operational planning process....

  5. Stool Culture

    Science.gov (United States)

    ... infections and may be identified with a stool culture. Some important examples include: Escherichia coli 0157:H7 and other toxin- ... the toxin-producing C. difficile will be performed. Examples of other less common causes include: ... of stool cultures that are reported as negative usually reflect the ...

  6. Boosting Culture

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    Culture makes up an indispensable part of our lives, just like material comfort. It is thought of as an important source of a nation’s vitality and creativity, and constitutes a key factor uniting the nation,while making it distinctive from other countries. It is also said culture is a productiv

  7. Hydroponics or soilless culture

    Science.gov (United States)

    Chapman, H. D.

    1963-01-01

    Historically, hydroponics is not a new field; plant physiologists have known and used it for some 100 years. Inevitably, some enthusiasts got carried away.Claims were made of enormous potential yields; skyscraper tops were said to be capable of producing enough food for all of their occupants; and closets, basements, garages, etc. were wishfully converted into fields for hydroponic culture. Numerous publications on the subject appeared during this period. Basic requirements for hydropinc techniques are given along with examples of where soilless culture has been used commercially.

  8. Internet culture

    CERN Document Server

    Porter, David

    2013-01-01

    The internet has recently grown from a fringe cultural phenomenon to a significant site of cultural production and transformation. Internet Culture maps this new domain of language, politics and identity, locating it within the histories of communication and the public sphere. Internet Culture offers a critical interrogation of the sustaining myths of the virtual world and of the implications of the current mass migration onto the electronic frontier. Among the topics discussed in Internet Culture are the virtual spaces and places created by the citizens of the Net and their claims to the hotly contested notion of "virtual community"; the virtual bodies that occupy such spaces; and the desires that animate these bodies. The contributors also examine the communication medium behind theworlds of the Net, analyzing the rhetorical conventions governing online discussion, literary antecedents,and potential pedagogical applications.

  9. Avatar Culture

    DEFF Research Database (Denmark)

    Koda, Tomoko; Ishida, Toru; Rehm, Matthias

    2009-01-01

    and Western designers. The goals of the study were: (1) to investigate cultural differences in avatar expression evaluation and apply findings from psychological studies of human facial expression recognition, (2) to identify expressions and design features that cause cultural differences in avatar facial...... expression interpretation. The results of our study confirmed that (1) there are cultural differences in interpreting avatars’ facial expressions, and the psychological theory that suggests physical proximity affects facial expression recognition accuracy is also applicable to avatar facial expressions, (2......Avatars are increasingly used to express our emotions in our online communications. Such avatars are used based on the assumption that avatar expressions are interpreted universally among all cultures. This paper investigated cross-cultural evaluations of avatar expressions designed by Japanese...

  10. CULTURAL TOURISM

    Directory of Open Access Journals (Sweden)

    Dana POP

    2016-07-01

    Full Text Available In this paper we will try to analyse the cultural tourism. We will start by referring to the complex concepts of tourism and culture and to the synergies existing between them. We will define cultural tourism and present its appearance and evolution as well as its importance as a modern form of tourism. We will present the various types of cultural tourism with their characteristics and the specific features of cultural tourists according to their interests. We will also mention that there are advantages and disadvantages for any kind of tourism depending on the position – local communities, companies or tourists. For the future we will refer to the new partnership between UNWTO and UNESCO.

  11. 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)

  12. Diagnostic techniques

    International Nuclear Information System (INIS)

    Berquist, T.H.; Bender, C.E.; James, E.M.; Brown, M.L.; McLeod, R.A.; Broderick, D.F.; Welch, T.J.

    1989-01-01

    Proper application of imaging procedures is essential to obtain needed information for diagnosis and therapy planning in patients with suspected foot and/or ankle pathology. This paper provides basic background data for the numerous imaging techniques

  13. Paramilitary Culture.

    Science.gov (United States)

    Gibson, James William

    1989-01-01

    Identifies the movie, "Rambo," and "Soldier of Fortune" magazine as artifacts of "paramilitary culture." Contends that they are a social phenomenon which helps legitimate the United States government's rapid escalation of military forces. (MS)

  14. Rectal culture

    Science.gov (United States)

    ... have an infection of the rectum, such as gonorrhea . It may also be done instead of a ... infection. This may be: Bacterial infection Parasitic enterocolitis Gonorrhea Sometimes a culture shows that you are a ...

  15. Cultural Years [SCRIPT 7 of 7]. "Music, the Arts and Society at the time of the Exposition Internationale des Arts et Techniques dans la Vie Moderne in Paris of 1837"\\ud [RADIO SERIES

    OpenAIRE

    Little, Jonathan D.

    1990-01-01

    Part of a seven-part radio series broadcast on Sundays at 10:30pm fortnightly from 1st April, 1990. Researched, written, presented and produced by Jonathan David Little for 3MBS-FM Fine Music Melbourne.\\ud \\ud CULTURAL YEARS - Series / Programme Description:\\ud \\ud “Cultural Years” was a seven-part radio series which discussed “Music, the Arts and Society around the time of seven of the great 19th- and 20th-century International Exhibitions” (see below). In examining the ideas which lay behin...

  16. Safety culture

    International Nuclear Information System (INIS)

    1991-01-01

    The response to a previous publication by the International Nuclear Safety Advisory Group (INSAG), indicated a broad international interest in expansion of the concept of Safety Culture, in such a way that its effectiveness in particular cases may be judged. This report responds to that need. In its manifestation, Safety Culture has two major components: the framework determined by organizational policy and by managerial action, and the response of individuals in working within and benefiting by the framework. 1 fig

  17. Decomposition techniques

    Science.gov (United States)

    Chao, T.T.; Sanzolone, R.F.

    1992-01-01

    Sample decomposition is a fundamental and integral step in the procedure of geochemical analysis. It is often the limiting factor to sample throughput, especially with the recent application of the fast and modern multi-element measurement instrumentation. The complexity of geological materials makes it necessary to choose the sample decomposition technique that is compatible with the specific objective of the analysis. When selecting a decomposition technique, consideration should be given to the chemical and mineralogical characteristics of the sample, elements to be determined, precision and accuracy requirements, sample throughput, technical capability of personnel, and time constraints. This paper addresses these concerns and discusses the attributes and limitations of many techniques of sample decomposition along with examples of their application to geochemical analysis. The chemical properties of reagents as to their function as decomposition agents are also reviewed. The section on acid dissolution techniques addresses the various inorganic acids that are used individually or in combination in both open and closed systems. Fluxes used in sample fusion are discussed. The promising microwave-oven technology and the emerging field of automation are also examined. A section on applications highlights the use of decomposition techniques for the determination of Au, platinum group elements (PGEs), Hg, U, hydride-forming elements, rare earth elements (REEs), and multi-elements in geological materials. Partial dissolution techniques used for geochemical exploration which have been treated in detail elsewhere are not discussed here; nor are fire-assaying for noble metals and decomposition techniques for X-ray fluorescence or nuclear methods be discussed. ?? 1992.

  18. Urine culture - catheterized specimen

    Science.gov (United States)

    Culture - urine - catheterized specimen; Urine culture - catheterization; Catheterized urine specimen culture ... urinary tract infections may be found in the culture. This is called a contaminant. You may not ...

  19. Cultural tourism and tourism cultures

    DEFF Research Database (Denmark)

    Ooi, Can-Seng

    Presenting a comprehensive and dynamic understanding of cultural tourism, this volume examines cultural mediators and how they help tourists appreciate foreign cultures. It also shows how tourism experiences are strategically crafted by mediators, the complexity of the mediation process, and how...... various products are mediated differently. A number of different products are investigated, including destination brand identities, "living" cultures and everyday life, art and history. The author illustrates his arguments by comparing the tourism strategies of Copenhagen and Singapore, and demonstrates...... how tourism is an agent for social change. The author also offers an original and refreshing way of understanding tourist behaviour through the concept of the "versatile tourist". The book's empirical cases and dialogic framework provide new and deep insights into tourism activities. In his...

  20. Cultured Trash, Not Trash Culture

    Directory of Open Access Journals (Sweden)

    Taufiqurrohman Taufiqurrohman

    2017-09-01

    Full Text Available As civilized creature, human actually can manage trash as well as possible although it is often stereotyped as a vain thing. This article gives the proof that trash can be cultured as well so that a society can take benefits from the existence of it. This article parses ways of orderly managing it at schools, in this case two schools in Jepara. The results say that trash can be cultured by having an organization to manage the Trash Bank at schools and to train students to classify and recycle trash then take advantage of it by selling the collected and the recycled trash. It makes trash have good transformation of values, repelling against the prior stereotype. Finally, by taking example from Trash Bank management at schools, human can have so cultured trash that they would not be trapped by trash culture.

  1. A Cultural Sexuality or a Sexual Culture?

    NARCIS (Netherlands)

    Vandermeersch, Patrick

    1990-01-01

    P. Vandermeersch, A Cultural Sexuality or a Sexual Culture? In: F. VAN DE VIJVER & G. HUTSCHEMAEKERS (ed.), The Investigation of Culture. Current Issues in Cultural Psychology, Tilburg, Tilburg University Press, 1990, 43-58.

  2. Neutron techniques

    International Nuclear Information System (INIS)

    Charlton, J.S.

    1986-01-01

    The way in which neutrons interact with matter such as slowing-down, diffusion, neutron absorption and moderation are described. The use of neutron techniques in industry, in moisture gages, level and interface measurements, the detection of blockages, boron analysis in ore feedstock and industrial radiography are discussed. (author)

  3. Cultural Resurrection

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    "Who are we?Where are we from?"Humans have been pondering these questions since the day they first came into being.One of the ways we preserve memories of the past is through our cul- tural heritage that has been passed on from generation to genera- tion.Intangible cultural heritage,as well as tangible cultural her- itage,is essential to the continuity of human civilization. Since the United Nations Educational,Scientific and Cultural Organization(UNESCO)unveiled the Masterpieces of the Oral and Intangible Heritage of Humanity in 2001,China has had Kunqu opera,Guqin and its music,the art of Uygur Muqam of Xinjiang and the traditional Mongolian folk song Long Song added to UNESCO’s protection list.It is now one of the coun-

  4. Japanese Shame Culture and American Guilt Culture

    Institute of Scientific and Technical Information of China (English)

    Lu Weijie

    2016-01-01

    Culture is an important factor contributing to the success of intercultural communication. In the east and west, there are many different cultures, among which Japanese shame culture and American guilt culture are two typical ones. Influenced by different cultures, these two countries have different characteristics, which reminds us that in intercultural communication culture should be paid much attention to.

  5. Mayan Culture

    DEFF Research Database (Denmark)

    Hervik, Peter Bent

    1992-01-01

    The social categories « Maya » and « mestizo » habe been applied to denote the Yucatec Mayan people in Mexico. The A. examines the cluster of perceived attributes (schemata) evoked by the terms and how they relate to each other. He shows that there is an incongruency between them along the lines ...... of local and academic categorization, which is an implication of the different social spaces in which they arise. In spite of the incongruency and the cultural plurality evoked by their usage, the A. argues that the people of Yucatec share a single culture....

  6. Information cultures

    DEFF Research Database (Denmark)

    Skouvig, Laura

    2017-01-01

    The purpose of this article is to suggest a genealogy of the concept of information beyond the 20th century. The article discusses how the concept of information culture might provide a way of formulating such a genealogic strategy. The article approaches this purpose by providing a general...... narrative of premodern information cultures, examining works on early-modern scholars and 18th century savants and discussion of what seems to be a Foucauldian rupture in the conceptualization of information in 19th century England. The findings of the article are situated in the thinking that a genealogy...... of information would reveal that information had specific purposes in specific settings....

  7. Astrophysical techniques

    CERN Document Server

    Kitchin, CR

    2013-01-01

    DetectorsOptical DetectionRadio and Microwave DetectionX-Ray and Gamma-Ray DetectionCosmic Ray DetectorsNeutrino DetectorsGravitational Radiation Dark Matter and Dark Energy Detection ImagingThe Inverse ProblemPhotographyElectronic ImagingScanningInterferometrySpeckle InterferometryOccultationsRadarElectronic ImagesPhotometryPhotometryPhotometersSpectroscopySpectroscopy SpectroscopesOther TechniquesAstrometryPolarimetrySolar StudiesMagnetometryComputers and The Internet.

  8. Presentation Technique

    International Nuclear Information System (INIS)

    Froejmark, M.

    1992-10-01

    The report presents a wide, easily understandable description of presentation technique and man-machine communication. General fundamentals for the man-machine interface are illustrated, and the factors that affect the interface are described. A model is presented for describing the operators work situation, based on three different levels in the operators behaviour. The operator reacts routinely in the face of simple, known problems, and reacts in accordance with predetermined plans in the face of more complex, recognizable problems. Deep fundamental knowledge is necessary for truly complex questions. Today's technical status and future development have been studied. In the future, the operator interface will be based on standard software. Functions such as zooming, integration of video pictures, and sound reproduction will become common. Video walls may be expected to come into use in situations in which several persons simultaneously need access to the same information. A summary of the fundamental rules for the design of good picture ergonomics and design requirements for control rooms are included in the report. In conclusion, the report describes a presentation technique within the Distribution Automation and Demand Side Management area and analyses the know-how requirements within Vattenfall. If different systems are integrated, such as geographical information systems and operation monitoring systems, strict demands are made on the expertise of the users for achieving a user-friendly technique which is matched to the needs of the human being. (3 figs.)

  9. Blood culture

    Science.gov (United States)

    ... There, it is placed in a special dish (culture). It is then watched to see if bacteria or other disease-causing germs grow. A gram stain may also ... any time the skin is broken) Alternative Names ... Charnot-Katsikas A. Specimen collection and handling for diagnosis of infectious diseases. In: McPherson RA, Pincus MR, eds. Henry's Clinical ...

  10. Cultural Usability

    DEFF Research Database (Denmark)

    Shi, Qingxin

    2007-01-01

    Culture has already played an important role in the global market. It not only affects products, but also impacts on usability evaluation methods. This project aims to examine in the established thinking aloud usability evaluation method (TA UEM), how does the evaluator build a supportive...

  11. Grindr Culture

    DEFF Research Database (Denmark)

    Shield, Andrew DJ

    2018-01-01

    intersections of sexuality with other socio-cultural categories such as race and migration background, but also gender and ability. I find that user experiences with exclusion and discrimination can be related to Grindr’s interface, such as its drop-down menus, the discourses circulated by Grindr users...

  12. Repolarization of hepatocytes in culture.

    Science.gov (United States)

    Talamini, M A; Kappus, B; Hubbard, A

    1997-01-01

    We have evaluated the biochemical, morphological, and functional redevelopment of polarity in freshly isolated hepatocytes cultured using a double layer collagen gel sandwich technique. Western blot analysis showed increased cellular levels of the cell adhesion protein uvomorulin as cultured hepatocytes repolarized. Immunofluorescence studies using antibodies against domain-specific membrane proteins showed polarity as early as 48 hours, although the pattern of the polymeric Immunoglobulin-A receptor (pIgA-R) differed from in vivo liver. Electron microscopy showed developing bile canaliculi at 1 day. However, the functional presence of tight junctions was absent at 1 day, but present at 5 days. We further showed functional polarity to be present at 4 days by documenting the ability of cultured hepatocytes to metabolize and excrete fluorescein diacetate into visible bile canaliculi. We conclude that hepatocytes cultured appropriately develop morphological and functional polarity. Hepatocyte culture is therefore a useful tool for the study of mechanisms responsible for the development of polarized function.

  13. Methods for plant molecular biology

    National Research Council Canada - National Science Library

    Weissbach, Arthur; Weissbach, Herbert

    1988-01-01

    .... Current techniques to carry out plant cell culture and protoplast formation are described as are methods for gene and organelle transfer. The detection of DNA and RNA viruses by molecular probes or ELISA assays and the cloning and transcription of viral RNA complete the volume.

  14. Astrophysical techniques

    Energy Technology Data Exchange (ETDEWEB)

    Kitchin, C R

    1984-01-01

    The subject is covered in chapters, entitled: detectors (optical and infrared detection; radio and microwave detection; X-ray and gamma-ray detection; cosmic ray detectors; neutrino detectors; gravitational radiation); imaging (photography; electronic imaging; scanning; interferometry; speckle interferometry; occultations; radar); photometry and photometers; spectroscopy and spectroscopes; other techniques (astrometry; polarimetry; solar studies; magnetometry). Appendices: magnitudes and spectral types of bright stars; north polar sequence; standard stars for the UBV photometric system; standard stars for the UVBY photometric system; standard stars for MK spectral types; standard stars for polarimetry; Julian date; catalogues; answers to the exercises.

  15. Llizarov technique

    International Nuclear Information System (INIS)

    Shankman, S.; Rosenberg, Z.S.; Frankel, V.; Golyakhovsky, V.

    1990-01-01

    This paper illustrates the radiographic manifestations of the Ilizarov distraction technique for the correction of short and deformed limbs. The radiographs of 130 patients who underwent Ilizarov distraction at 160 sites since December 1986 were reviewed retrospectively. Reasons for correction included fracture nonunion and malunion idiopathic leg length discrepancy, achondroplasia, and neurofibromatosis with tibial pseudarthrosis. Ninety patients were adults, and 40 were children. In order to assess cortical bone development during the fixation phase, CT of the regenerate bone was performed in 17 patients. Conventional tomograms were obtained in 20 patients for the evaluation of delayed or hypoplastic union at the distraction site

  16. Experimental Techniques

    International Nuclear Information System (INIS)

    Fournier, D.; Serin, L.

    1995-01-01

    Experimental techniques to be used in the new generation of high energy physics are presented. The emphasis is put on the new ATLAS and CMS detectors for the CERN LHC. For the most important elements of these detectors, a description of the underlying physics processes is given, sometimes with reference to comparable detectors used in the past. Some comparative global performances of the two detectors are also given, with reference to benchmark physics processes (detection of the Higgs boson in various mass regions, etc). (author)

  17. Grist and mills: on the cultural origins of cultural learning

    Science.gov (United States)

    Heyes, Cecilia

    2012-01-01

    Cumulative cultural evolution is what ‘makes us odd’; our capacity to learn facts and techniques from others, and to refine them over generations, plays a major role in making human minds and lives radically different from those of other animals. In this article, I discuss cognitive processes that are known collectively as ‘cultural learning’ because they enable cumulative cultural evolution. These cognitive processes include reading, social learning, imitation, teaching, social motivation and theory of mind. Taking the first of these three types of cultural learning as examples, I ask whether and to what extent these cognitive processes have been adapted genetically or culturally to enable cumulative cultural evolution. I find that recent empirical work in comparative psychology, developmental psychology and cognitive neuroscience provides surprisingly little evidence of genetic adaptation, and ample evidence of cultural adaptation. This raises the possibility that it is not only ‘grist’ but also ‘mills’ that are culturally inherited; through social interaction in the course of development, we not only acquire facts about the world and how to deal with it (grist), we also build the cognitive processes that make ‘fact inheritance’ possible (mills). PMID:22734061

  18. Talking Culture

    DEFF Research Database (Denmark)

    Holmgreen, Lise-Lotte

    When Danish businesses move production abroad, ‘culture’ is often seen as a huge challenge to the successful outcome of cross-border collaboration. Therefore, business leaders often seek information and guidelines of how to cope in the vast amount of literature on culture and intercultural...... communication. Much of this literature is based on functionalist approaches providing the dos and don’ts of intercultural encounters. This involves inter alia conceptualising ‘culture’ as a relatively fixed, homogeneous entity of values, attitudes and norms shared by members of a group, often leading readers...... to adopt dichotomised understandings and discourses about other cultures (see e.g. Hofstede 2001; Jandt 1998; Trompenaars & Hampden-Turner 1997). However, experience shows that the world in which intercultural encounters take place is not as simple and easy to categorise as these approaches may suggest...

  19. MARKETING CULTURAL

    Directory of Open Access Journals (Sweden)

    Claudia Gómez Ramírez

    2013-07-01

    Full Text Available Este artículo analiza la definición de "Marketing" Cultural y la adaptación y beneficios del "marketing" tradicional respecto al conjunto de manifestaciones artísticas de las diversas industrias involucradas en el sector cultural o artístico; asimismo, se desagregan los conceptos básicos que lo componen como factor de éxito en dichas empresas. Se hace uso de la exposición de casos específicos para ilustrar la articulación de estos dos conceptos aparentemente contrapuestos, cultura y "marketing", y registra algunas reflexiones para que el lector se involucre en la construcción del concepto aquí presentado.

  20. A microculture technique for rat lymphocyte transformation.

    Science.gov (United States)

    Lindsay, V J; Allardyce, R A

    1979-01-01

    We report the development of an economical microculture technique suitable for measuring rat lymphocyte response to mitogens and in mixed lymphocyte reactions. The effects of varying culture conditions, i.e. source of serum, addition and concentration of 2-mercaptoethanol, mitogen concentrations, culture incubation times, absorption of serum, lymphocyte numbers and microtitre plate well shape are described.

  1. Laboratory Workflow Analysis of Culture of Periprosthetic Tissues in Blood Culture Bottles.

    Science.gov (United States)

    Peel, Trisha N; Sedarski, John A; Dylla, Brenda L; Shannon, Samantha K; Amirahmadi, Fazlollaah; Hughes, John G; Cheng, Allen C; Patel, Robin

    2017-09-01

    Culture of periprosthetic tissue specimens in blood culture bottles is more sensitive than conventional techniques, but the impact on laboratory workflow has yet to be addressed. Herein, we examined the impact of culture of periprosthetic tissues in blood culture bottles on laboratory workflow and cost. The workflow was process mapped, decision tree models were constructed using probabilities of positive and negative cultures drawn from our published study (T. N. Peel, B. L. Dylla, J. G. Hughes, D. T. Lynch, K. E. Greenwood-Quaintance, A. C. Cheng, J. N. Mandrekar, and R. Patel, mBio 7:e01776-15, 2016, https://doi.org/10.1128/mBio.01776-15), and the processing times and resource costs from the laboratory staff time viewpoint were used to compare periprosthetic tissues culture processes using conventional techniques with culture in blood culture bottles. Sensitivity analysis was performed using various rates of positive cultures. Annualized labor savings were estimated based on salary costs from the U.S. Labor Bureau for Laboratory staff. The model demonstrated a 60.1% reduction in mean total staff time with the adoption of tissue inoculation into blood culture bottles compared to conventional techniques (mean ± standard deviation, 30.7 ± 27.6 versus 77.0 ± 35.3 h per month, respectively; P < 0.001). The estimated annualized labor cost savings of culture using blood culture bottles was $10,876.83 (±$337.16). Sensitivity analysis was performed using various rates of culture positivity (5 to 50%). Culture in blood culture bottles was cost-effective, based on the estimated labor cost savings of $2,132.71 for each percent increase in test accuracy. In conclusion, culture of periprosthetic tissue in blood culture bottles is not only more accurate than but is also cost-saving compared to conventional culture methods. Copyright © 2017 American Society for Microbiology.

  2. Cultural Heritage communication technology

    Directory of Open Access Journals (Sweden)

    Elena Ippoliti

    2011-12-01

    Full Text Available This magazine issue is about the relationship between digital techniques and the communication of cultural heritage and specifically aims at portraying how the interest and implications of these two things are widespread. Without trying to go too in depth, various points of view have been compared, each taken from different articles presenting a wide range of possible approaches on the subject of creating a wealth of information on cultural heritage and how it can be made available to the public without difficulty. Therefore, this issue wants to create a forum for a many-sided comparison built on a wealth of experience and opinions of different authors. In this way the abundance and versatility of the contributing professions (architects, archaeologists, engineers, mathematicians, graphic designers, artists, video producers, digital experts, 3D graphic designers, critics, directors, etc. has given life to a precious blend of know-how, which is without doubt enhanced by present-day digital technology.

  3. Cultural Robotics: The Culture of Robotics and Robotics in Culture

    Directory of Open Access Journals (Sweden)

    Hooman Samani

    2013-12-01

    Full Text Available In this paper, we have investigated the concept of “Cultural Robotics” with regard to the evolution of social into cultural robots in the 21st Century. By defining the concept of culture, the potential development of a culture between humans and robots is explored. Based on the cultural values of the robotics developers, and the learning ability of current robots, cultural attributes in this regard are in the process of being formed, which would define the new concept of cultural robotics. According to the importance of the embodiment of robots in the sense of presence, the influence of robots in communication culture is anticipated. The sustainability of robotics culture based on diversity for cultural communities for various acceptance modalities is explored in order to anticipate the creation of different attributes of culture between robots and humans in the future.

  4. Industrial technique

    International Nuclear Information System (INIS)

    Robertson, D.H.

    1975-01-01

    Techniques for selectively extracting and storing only krypton and xenon in the waste gases that are released from a pressurized water nuclear power reactor are described. The illustrative fluorocarbon absorption system has three separation stages: an initial gas-fluorocarbon absorber, a flash chamber and fractionator for segregating all of the absorbed gases from the loaded absorber stage fluorocarbon (save for the krypton and xenon), and a stripper that receives the partially loaded fluorocarbon liquid directly from the fractionator in order to separate only the krypton and xenon. A molecular sieve filter dries the input process gas, a cartridge type solvent filter is used to remove radiation degradation products from the loaded liquid that flows from the absorber, a cold trap gas drier is provided to remove residual solvent vapor from the separated krypton and xenon, and radiation detectors automatically activate valves to establish safe conditions in the event of an accident or plant failure. (U.S.)

  5. Experimental techniques

    International Nuclear Information System (INIS)

    Roussel-Chomaz, P.

    2007-01-01

    This lecture presents the experimental techniques, developed in the last 10 or 15 years, in order to perform a new class of experiments with exotic nuclei, where the reactions induced by these nuclei allow to get information on their structure. A brief review of the secondary beams production methods will be given, with some examples of facilities in operation or under project. The important developments performed recently on cryogenic targets will be presented. The different detection systems will be reviewed, both the beam detectors before the targets, and the many kind of detectors necessary to detect all outgoing particles after the reaction: magnetic spectrometer for the heavy fragment, detection systems for the target recoil nucleus, γ detectors. Finally, several typical examples of experiments will be detailed, in order to illustrate the use of each detector either alone, or in coincidence with others. (author)

  6. Combinatorial techniques

    CERN Document Server

    Sane, Sharad S

    2013-01-01

    This is a basic text on combinatorics that deals with all the three aspects of the discipline: tricks, techniques and theory, and attempts to blend them. The book has several distinctive features. Probability and random variables with their interconnections to permutations are discussed. The theme of parity has been specially included and it covers applications ranging from solving the Nim game to the quadratic reciprocity law. Chapters related to geometry include triangulations and Sperner's theorem, classification of regular polytopes, tilings and an introduction to the Eulcidean Ramsey theory. Material on group actions covers Sylow theory, automorphism groups and a classification of finite subgroups of orthogonal groups. All chapters have a large number of exercises with varying degrees of difficulty, ranging from material suitable for Mathematical Olympiads to research.

  7. Western´s decline, culture tragedy and Titan´s rise: genealogical elements of the discussion of technique in Heidegger Declínio do ocidente, tragédia da cultura e ascensão dos Titãs: elementos genealógicos da discussão da técnica em Heidegger

    Directory of Open Access Journals (Sweden)

    Francisco Rüdiger

    2008-06-01

    Full Text Available The article intends to signalize the debts of the Heideggerian meditation on technique from the ideas of three thinkers: Simmel, Spengler and Ernst Jünger. After sketches up the context in which they arisen, the article focuses in their main thesis to confront them with the way they are appropriate by Heidegger. The article concludes stressing the transcendent character of the points in it analyzed to the enlightenment of the question of technique according to Heidegger. Keywords: Heidegger and the question of technique. Technique and modern culture. Quer-se com o texto apontar as dívidas da reflexão heideggeriana sobre a técnica moderna para com as idéias de três pensadores: Simmel, Spengler e Ernst Jünger. Depois de breve contextualização do problema, o essencial das teses de cada um é posto à luz e confrontado com sua apropriação por parte de Heidegger. Indicando conexões e rupturas, conclui-se chamando à atenção para o interesse transcendente dos pontos em questão. Palavras-chave: Heidegger e a questão da técnica. Técnica e cultura moderna.

  8. La arquitectura cultural. / Cultural architecture.

    Directory of Open Access Journals (Sweden)

    Lobos, Jorge

    2004-12-01

    Full Text Available Este artículo releva la Arquitectura Cultural, que es plural y diversa en términos estéticos y conceptuales. Sugiere la apertura a otras dinámicas de comprensión de lo arquitectónico, a otras lógicas de construcción de las ciudades. Para aclarar el concepto se expone un breve ejemplo práctico./This article points out the "cultural architecture", which is plural and diverse in concept and aesthetic terms. It suggests the opening to other dinamics of comprehention of the architectural issue and the building of cities. The article presents a brief empirical example that clarifies the conceptual approach.

  9. Cultural Robotics: The Culture of Robotics and Robotics in Culture

    OpenAIRE

    Hooman Samani; Elham Saadatian; Natalie Pang; Doros Polydorou; Owen Noel Newton Fernando; Ryohei Nakatsu; Jeffrey Tzu Kwan Valino Koh

    2013-01-01

    In this paper, we have investigated the concept of “Cultural Robotics” with regard to the evolution of social into cultural robots in the 21st Century. By defining the concept of culture, the potential development of a culture between humans and robots is explored. Based on the cultural values of the robotics developers, and the learning ability of current robots, cultural attributes in this regard are in the process of being formed, which would define the new concept of cultural robotics. Ac...

  10. Digital Preservation of Cultural Heritage

    NARCIS (Netherlands)

    Peters, Nonja; Marinova, Dora; van Faassen, M.; Stasiuk, Glen; Zacher, L.W.

    2017-01-01

    The focus of this chapter is the state of the art of digitisation of cultural heritage in Australian archives and libraries from a comparative perspective. Globalisation, mobility and the new techniques that spin off from the digital age bring about new possibilities that stimulate and enhance our

  11. Black Culture

    Directory of Open Access Journals (Sweden)

    Angela Khristin Brown

    2013-07-01

    Full Text Available The migration of blacks in North America through slavery became united.  The population of blacks past downs a tradition of artist through art to native born citizens. The art tradition involved telling stories to each generation in black families. The black culture elevated by tradition created hope to determine their personal freedom to escape from poverty of enslavement and to establish a way of life through tradition. A way of personal freedoms was through getting a good education that lead to a better foundation and a better way of life.

  12. Culture Jamming Versus Popular Culture

    Directory of Open Access Journals (Sweden)

    Leonardia Acynthia Putri

    2013-11-01

    Full Text Available Abstract: This literature study researched Adbusters, the anti-commercial organization, and described the organization’s activities and media usage, mainly in the period of 2007-2010, which critized the populer culture. Adbusters is an organization which performs “Culture Jamming”; a rebellious act reacting towards commercialism domination in many aspects including popular culture. Compared to other similar organizations, Adbusters has been executing more various activisms using several media which other organizations do not use. This study used the Adbusters’ official website and blogs as main data sources. The data of Adbusters’ activities and media usage were categorized and analyzed, thus the tendency of its development can be described. This study also analyzed Adbusters’ activity using Media Hegemony Theory and Political Economy Media Theory. The media has been dominated by a certain group that owns politic and economic power, so the information flow has been dominated by them. Media and its contents have been commercialized, thus capitalism and commercialism have been considered as a common system that should run the world. Adbusters has been trying to stop the domination and change the society’s way of thinking into a more critical way of thinking.   Abstrak: Studi literatur ini meneliti tentang Adbusters, sebuah organisasi anti komersial, dengan mendeskripsikan aktivitas serta penggunaan media organisasi tersebut dari tahun 2007-2010 dalam mengkritisi budaya populer. Adbusters adalah organisasi yang melakukan Culture Jamming, aksi perlawanan terhadap dominasi komersialisme di segala aspek termasuk popular culture. Dibandingkan dengan organisasi lain yang serupa, aktivitas Adbusters lebih bervariasi dan menggunakan media-media yang tidak biasa digunakan organisasi lain. Penelitian ini menggunakan situs online resmi Adbusters sebagai sumber data utama. Data mengenai aktivitas dan

  13. NAIL SAMPLING TECHNIQUE AND ITS INTERPRETATION

    Directory of Open Access Journals (Sweden)

    TZAR MN

    2011-01-01

    Full Text Available The clinical suspicion of onychomyosis based on appearance of the nails, requires culture for confirmation. This is because treatment requires prolonged use of systemic agents which may cause side effects. One of the common problems encountered is improper nail sampling technique which results in loss of essential information. The unfamiliar terminologies used in reporting culture results may intimidate physicians resulting in misinterpretation and hamper treatment decision. This article provides a simple guide on nail sampling technique and the interpretation of culture results.

  14. Industrial technique

    International Nuclear Information System (INIS)

    Jabsen, F.S.

    1975-01-01

    An illustrative embodiment of the invention discloses a technique for disassembling a nuclear reactor fuel element without destroying the individual fuel pins and other structural components from which the element is assembled. A traveling bridge and trolley that span a water-filled spent fuel storage pool support a strongback. The strongback is under water and provides a working surface on which the spent fuel element is placed for inspection and for the manipulation that is associated with disassembly and assembly. To remove, in a non-destructive manner, the grids that hold the fuel pins in the proper relative positions within the element, bars are inserted through apertures in the grids with the aid of special tools. These bars are rotated to flex the adjacent grid walls and, in this way relax the physical engagement between protruding portions of the grid walls and the associated fuel pins. With the grid structure so flexed to relax the physical grip on the individual fuel pins, these pins can be withdrawn for inspection or replacement as necessary without imposing a need to destroy fuel element components. (U.S.)

  15. Culture collections.

    Science.gov (United States)

    Smith, David

    2012-01-01

    Culture collections no matter their size, form, or institutional objectives play a role in underpinning microbiology, supplying the resources for study, innovation, and discovery. Their basic roles include providing a mechanism for ex situ conservation of organisms; they are repositories for strains subject to publication, taking in safe, confidential, and patent deposits from researchers. They supply strains for use; therefore, the microorganisms provided must be authentic and preserved well, and any associated information must be valid and sufficient to facilitate the confirmation of their identity and to facilitate their use. The organisms must be collected in compliance with international conventions, international and national legislation and distributed to users indicating clearly the terms and conditions under which they are received and can be used. Collections are harmonizing approaches and characterizing strains to meet user needs. No one single collection can carry out this task alone, and therefore, it is important that output and strategy are coordinated to ensure culture collections deliver the basic resources and services microbiological innovation requires. This chapter describes the types of collection and how they can implement quality management systems and operate to deliver their basic functions. The links to information sources given not only provide support for the practitioners within collections but also provide guidance to users on accessing the huge resource available and how they can help ensure microbiology has the resources and a solid platform for future development. Copyright © 2012 Elsevier Inc. All rights reserved.

  16. Cultural Heritage Education for Intercultural Communication

    OpenAIRE

    Kokko, Sirpa; Kyritsi, Anna

    2012-01-01

    In this paper, cultural heritage is considered as an important aspect of intercultural communication and social cohesion, both in local communities as well as on the European level. In European societies of today, the role of the cultural heritage of arts and crafts is under discussion. Attention has turned to the importance of conserving and developing traditional knowledge and techniques. On the basis of this and the practical experiences from craft and cultural heritage projects in Finland...

  17. Culture in Southeast Asian Language Classes.

    Science.gov (United States)

    Liem, Nguyen Dang

    A view of the status of Southeast Asian language programs in American schools leads the author to comment on five interrelated issues. They include: (1) the importance of Southeast Asian language and culture teaching and learning, (2) integrating culture in Southeast Asian language classes, (3) teaching techniques, (4) staffing, and (5)…

  18. Techniques de combustion Combustin Techniques

    Directory of Open Access Journals (Sweden)

    Perthuis E.

    2006-11-01

    Full Text Available L'efficacité d'un processus de chauffage par flamme est étroitement liée à la maîtrise des techniques de combustion. Le brûleur, organe essentiel de l'équipement de chauffe, doit d'une part assurer une combustion complète pour utiliser au mieux l'énergie potentielle du combustible et, d'autre part, provoquer dans le foyer les conditions aérodynamiques les plus propices oux transferts de chaleur. En s'appuyant sur les études expérimentales effectuées à la Fondation de Recherches Internationales sur les Flammes (FRIF, au Groupe d'Étude des Flammes de Gaz Naturel (GEFGN et à l'Institut Français du Pétrole (IFP et sur des réalisations industrielles, on présente les propriétés essentielles des flammes de diffusion aux combustibles liquides et gazeux obtenues avec ou sans mise en rotation des fluides, et leurs répercussions sur les transferts thermiques. La recherche des températures de combustion élevées conduit à envisager la marche à excès d'air réduit, le réchauffage de l'air ou son enrichissement à l'oxygène. Par quelques exemples, on évoque l'influence de ces paramètres d'exploitation sur l'économie possible en combustible. The efficiency of a flame heating process is closely linked ta the mastery of, combustion techniques. The burner, an essential element in any heating equipment, must provide complete combustion sa as to make optimum use of the potential energy in the fuel while, at the same time, creating the most suitable conditions for heat transfers in the combustion chamber. On the basis of experimental research performed by FRIF, GEFGN and IFP and of industrial achievements, this article describesthe essential properties of diffusion flames fed by liquid and gaseous fuels and produced with or without fluid swirling, and the effects of such flames on heat transfers. The search for high combustion temperatures means that consideration must be given to operating with reduced excess air, heating the air or

  19. Microalgae harvesting techniques: A review.

    Science.gov (United States)

    Singh, Gulab; Patidar, S K

    2018-07-01

    Microalgae with wide range of commercial applications have attracted a lot of attention of the researchers in the last few decades. However, microalgae utilization is not economically sustainable due to high cost of harvesting. A wide range of solid - liquid separation techniques are available for microalgae harvesting. The techniques include coagulation and flocculation, flotation, centrifugation and filtration or a combination of various techniques. Despite the importance of harvesting to the economics and energy balance, there is no universal harvesting technique for microalgae. Therefore, this review focuses on assessing technical, economical and application potential of various harvesting techniques so as to allow selection of an appropriate technology for cost effectively harvesting of microalgae from their culture medium. Various harvesting and concentrating techniques of microalgae were reviewed to suggest order of suitability of the techniques for four main microalgae applications i.e biofuel, human and animal food, high valued products, and water quality restoration. For deciding the order of suitability, a comparative analysis of various harvesting techniques based on the six common criterions (i.e biomass quality, cost, biomass quantity, processing time, species specific and toxicity) has been done. Based on the order of various techniques vis-a-vis various criteria and preferred order of criteria for various applications, order of suitability of harvesting techniques for various applications has been decided. Among various harvesting techniques, coagulation and flocculation, centrifugation and filtration were found to be most suitable for considered applications. These techniques may be used alone or in combination for increasing the harvesting efficiency. Copyright © 2018 Elsevier Ltd. All rights reserved.

  20. Mindfulness in cultural context.

    Science.gov (United States)

    Kirmayer, Laurence J

    2015-08-01

    Mindfulness meditation and other techniques drawn from Buddhism have increasingly been integrated into forms of psychotherapeutic intervention. In much of this work, mindfulness is understood as a mode of awareness that is present-centered and nonevaluative. This form of awareness is assumed to have intrinsic value in promoting positive mental health and adaptation by interrupting discursive thoughts that give rise to suffering. However, in the societies where it originated, mindfulness meditation is part of a larger system of Buddhist belief and practice with strong ethical and moral dimensions. Extracting techniques like mindfulness meditation from the social contexts in which they originate may change the nature and effects of the practice. The papers in this issue of Transcultural Psychiatry explore the implications of a cultural and contextual view of mindfulness for continued dialogue between Buddhist thought and psychiatry. This introductory essay considers the meanings of mindfulness meditation in cultural context and the uses of mindfulness as a therapeutic intervention in contemporary psychiatry and psychology. © The Author(s) 2015.

  1. Hispanic Culture and Relational Cultural Theory

    Science.gov (United States)

    Ruiz, Elizabeth

    2005-01-01

    Traditional personality theories do not consider the impact of culture on personality development. Yet, to provide culturally relevant services to the increasing Hispanic population in the U.S., more culturally relevant theories must be identified. This paper presents Relational Cultural Theory (RCT) as an alternative model to understanding…

  2. Academic Culture and Campus Culture of Universities

    Science.gov (United States)

    Shen, Xi; Tian, Xianghong

    2012-01-01

    Academic culture of universities mainly consists of academic outlooks, academic spirits, academic ethics and academic environments. Campus culture in a university is characterized by individuality, academic feature, opening, leading, variety and creativity. The academic culture enhances the construction of campus culture. The campus culture…

  3. Culture-lovers and Culture-leavers

    NARCIS (Netherlands)

    Frank Huysmans; Andries van den Broek; Jos de Haan

    2005-01-01

    Who are the people in the Netherlands with an active interest in cultural heritage and the performing arts, and who prefer to leave these forms of culture alone? Have the size and composition of the groups of 'culture-lovers' and 'culture-leavers' changed since the end of the 1970s? These are the

  4. FROM CULTURAL IMPOTENCE TO CULTURAL AMPUTATION

    Directory of Open Access Journals (Sweden)

    Sukhanov Vyacheslav Vladimirovich

    2013-02-01

    Full Text Available Cultural space of any state is formed by a population that is within its borders. In this article, the author introduces a new cultural definitions «cultural impotence» and «cultural amputation», justifying their use, both in terms of population of the Russian Federation and the European Union and America. The article analyzes the state of society and the cultural factors that influence the development of society in Russia, there are options to bring the country out of a deep cultural crisis. Also established a close relationship between the domestic policy of the state and development of culture.

  5. INNOVATIONS IN EQUIPMENT AND TECHNIQUES FOR THE BIOLOGY TEACHING LABORATORY.

    Science.gov (United States)

    BARTHELEMY, RICHARD E.; AND OTHERS

    LABORATORY TECHNIQUES AND EQUIPMENT APPROPRIATE FOR TEACHING BIOLOGICAL SCIENCE CURRICULUM STUDY BIOLOGY ARE EMPHASIZED. MAJOR CATEGORIES INCLUDE (1) LABORATORY FACILITIES, (2) EQUIPMENT AND TECHNIQUES FOR CULTURE OF MICRO-ORGANISMS, (3) LABORATORY ANIMALS AND THEIR HOUSING, (4) TECHNIQUES FOR STUDYING PLANT GROWTH, (5) TECHNIQUES FOR STUDYING…

  6. Cultural Competence and the Operational Commander: Moving Beyond Cultural Awareness into Culture-Centric Warfare

    National Research Council Canada - National Science Library

    Karcanes, James A

    2007-01-01

    .... Understanding the different levels of cultural awareness -- cultural consideration, cultural understanding, and cultural competence -- will help usher in a new focus on culture-centric warfare...

  7. CONNECTION BETWEEN ECONOMICS, CULTURE AND CULTURAL DIPLOMACY

    Directory of Open Access Journals (Sweden)

    Agil Valiyev

    2017-09-01

    Full Text Available Today, culture is one of the main feeble factors of economic development.  The leading role of culture in economic development should be argued as multiplied: so, on firstly, as domestic value, on secondly, as a main factor of regional economic development advanced to raised gravity of different regions for residents, tourists and investors, on thirdly, as major parameters of social development based on tolerance, creativity and knowledge. To the different international experiences, culture is main part of economic development in our life. Cultural diversities are combined into a main reason economic development model. The article consist of explainations about the understanding of culture, cultural diplomacy and economics, approach on conflicts between culture and economics, to find how affecting of culture to economic development, the role of culture in economic development of Azerbaijan. The article can be considered as a useful resource  for experts and researchers conducting research in this field.

  8. Cultural Humility and Hospital Safety Culture.

    Science.gov (United States)

    Hook, Joshua N; Boan, David; Davis, Don E; Aten, Jamie D; Ruiz, John M; Maryon, Thomas

    2016-12-01

    Hospital safety culture is an integral part of providing high quality care for patients, as well as promoting a safe and healthy environment for healthcare workers. In this article, we explore the extent to which cultural humility, which involves openness to cultural diverse individuals and groups, is related to hospital safety culture. A sample of 2011 hospital employees from four hospitals completed measures of organizational cultural humility and hospital safety culture. Higher perceptions of organizational cultural humility were associated with higher levels of general perceptions of hospital safety, as well as more positive ratings on non-punitive response to error (i.e., mistakes of staff are not held against them), handoffs and transitions, and organizational learning. The cultural humility of one's organization may be an important factor to help improve hospital safety culture. We conclude by discussing potential directions for future research.

  9. MICROENCAPSULATION-THE FUTURE OF PROBIOTIC CULTURES

    Directory of Open Access Journals (Sweden)

    Tawheed Amin

    2013-08-01

    Full Text Available In the recent past, there has been an explosion of probiotic cultures based health products in Indian markets. The survival of the probiotic bacteria in gastro-intestinal gut is questionable, because of the poor survival of probiotic bacteria in these products. Basically the viability of probiotic cultures is very weak in these food products. Probiotic based products are health potentiators and are associated with many health benefits. Microencapsulation of the probiotic cultures is one of the recent, demanded and highly efficient techniques. Among the different approaches proposed to improve the survival of probiotics during food manufacturing process and passage in the upper part of gastrointestinal tratct (GI tract, microencapsulation has received considerable attention. Encapsulated probiotic cultures have longer shelf life of the products. This microencapsulation technology is used to maintain the viability of probiotic bacteria during food product processing and storage. This article reviews the principles, techniques and need for microencapsulation of probiotic cultures.

  10. Teaching Culture Through Films

    Institute of Scientific and Technical Information of China (English)

    徐婷

    2016-01-01

    Cultural teaching is an issue which is associated with complexity and paradox and also it is a big challenge for faculty. Teaching culture through films has become an important way of cross-cultural teaching This paper focuses on the reasons for teaching culture through films, the value and how it works. And finally it leads out the prospects of cultural teaching through films.

  11. Transient foreign gene expression in chloroplasts of cultured tobacco cells after biolistic delivery of chloroplast vectors.

    Science.gov (United States)

    Daniell, H; Vivekananda, J; Nielsen, B L; Ye, G N; Tewari, K K; Sanford, J C

    1990-01-01

    Expression of chloramphenicol acetyltransferase (cat) by suitable vectors in chloroplasts of cultured tobacco cells, delivered by high-velocity microprojectiles, is reported here. Several chloroplast expression vectors containing bacterial cat genes, placed under the control of either psbA promoter region from pea (pHD series) or rbcL promoter region from maize (pAC series) have been used in this study. In addition, chloroplast expression vectors containing replicon fragments from pea, tobacco, or maize chloroplast DNA have also been tested for efficiency and duration of cat expression in chloroplasts of tobacco cells. Cultured NT1 tobacco cells collected on filter papers were bombarded with tungsten particles coated with pUC118 (negative control), 35S-CAT (nuclear expression vector), pHD312 (repliconless chloroplast expression vector), and pHD407, pACp18, and pACp19 (chloroplast expression vectors with replicon). Sonic extracts of cells bombarded with pUC118 showed no detectable cat activity in the autoradiograms. Nuclear expression of cat reached two-thirds of the maximal 48 hr after bombardment and the maximal at 72 hr. Cells bombarded with chloroplast expression vectors showed a low level of expression until 48 hr of incubation. A dramatic increase in the expression of cat was observed 24 hr after the addition of fresh medium to cultured cells in samples bombarded with pHD407; the repliconless vector pHD312 showed about 50% of this maximal activity. The expression of nuclear cat and the repliconless chloroplast vector decreased after 72 hr, but a high level of chloroplast cat expression was maintained in cells bombarded with pHD407. Organelle-specific expression of cat in appropriate compartments was checked by introducing various plasmid constructions into tobacco protoplasts by electroporation. Although the nuclear expression vector 35S-CAT showed expression of cat, no activity was observed with any chloroplast vectors.

  12. Exploring Culture : Exercises, Stories and Synthetic Cultures

    NARCIS (Netherlands)

    Hofstede, G.J.

    2002-01-01

    A unique training book containing over 100 culture awareness exercises, dialogues, stories incidents and simulations that bring to life Geert Hofstede's five dimensions of culture. These dimensions are: power distance, collectivism versus individualism, femininity versus masculinity, uncertainly

  13. Dehistoricized Cultural Identity and Cultural Othering

    Science.gov (United States)

    Weiguo, Qu

    2013-01-01

    The assumption that each culture has its own distinctive identity has been generally accepted in the discussion of cultural identities. Quite often identity formation is not perceived as a dynamic and interactive ongoing process that engages other cultures and involves change in its responses to different challenges at different times. I will…

  14. Cultural Understanding Through Cross-Cultural Analysis.

    Science.gov (United States)

    Briere, Jean-Francois

    1986-01-01

    A college course used an explicit intercultural approach and collective research activities to compare French and American cultures and to examine the reasons for cultural attitudes and culture conflict. Class assignments dealt with contrastive analyses of American and French institutions like advertising, cinema, feminism, etc. (MSE)

  15. NAIL SAMPLING TECHNIQUE AND ITS INTERPRETATION

    OpenAIRE

    TZAR MN; LEELAVATHI M

    2011-01-01

    The clinical suspicion of onychomyosis based on appearance of the nails, requires culture for confirmation. This is because treatment requires prolonged use of systemic agents which may cause side effects. One of the common problems encountered is improper nail sampling technique which results in loss of essential information. The unfamiliar terminologies used in reporting culture results may intimidate physicians resulting in misinterpretation and hamper treatment decision. This article prov...

  16. Microalgal Culture Collection Transfers

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The Milford Microalgal culture Collection holds over 200 live cultures representing 13 classes of of algae. The cultures are maintained in three different growing...

  17. Routine sputum culture

    Science.gov (United States)

    Sputum culture ... There, it is placed in a special dish (culture). It is then watched to see if bacteria ... Elsevier; 2018:chap 36. Chernecky CC, Berger BJ. Culture, routine. In: Chernecky CC, Berger BJ, eds. Laboratory ...

  18. Lymph node culture

    Science.gov (United States)

    Culture - lymph node ... or viruses grow. This process is called a culture. Sometimes, special stains are also used to identify specific cells or microorganisms before culture results are available. If needle aspiration does not ...

  19. Cross-cultural awareness

    OpenAIRE

    БУРЯК Н.Ю.

    2016-01-01

    The article deals with the importance of cultural awareness for businesspeople when they go abroad. It also gives some cultural advice and factors which are thought to be the most important in creating a culture.

  20. Teaching Culturally Diverse Students.

    Science.gov (United States)

    Correa, Vivian; Tulbert, Beth

    1991-01-01

    Characteristics of culturally diverse students are discussed in terms of language, culture, and socioeconomic factors. Meeting the educational needs of culturally diverse students can involve interactive teaming of professionals; parent involvement; and providing appropriate services, assessment, curriculum, and instruction. (JDD)

  1. Cerebrospinal fluid culture

    Science.gov (United States)

    ... Alternative Names Culture - CSF; Spinal fluid culture; CSF culture Images Pneumococci organism References Karcher DS, McPherson RA. Cerebrospinal, synovial, serous body fluids, and alternative specimens. In: McPherson RA, Pincus ...

  2. Organizations, projects and culture

    NARCIS (Netherlands)

    J. van Cleeff; Pieter van Nispen tot Pannerden

    2016-01-01

    Purpose: to explore and demonstrate the effects of organizational culture on projects, in particular project culture and project management style. Methodology/approach: descriptive and explorative; through students’ groups. Findings: the cultural relationship between organizations, their projects

  3. A study of the process of synchronisation and micronucleation in Beta vulgaris and the monitoring of an isolation procedure for micro-nuclei and micro-protoplasts by confocal laser scanning microscopy and flow cytometry

    NARCIS (Netherlands)

    Famelaer, I.; Verhoeven, H.A.; Dijkhuis, P.; Ramulu, K.S.

    2007-01-01

    The process of synchronization and micro-nuclei induction in a suspension culture of Beta vulgaris, was induced by the sequential treatment with the DNA-synthesis inhibitor aphidicolin (30 mu M, 24 h) and the spindle-toxin amiprophos-methyl (32 mu M, 24 h). Mitotic arrest of divisions, spreading of

  4. Culture, Liberty and Happiness

    OpenAIRE

    Ura, Karma

    2007-01-01

    The author's intention here is to explore mainly the relationship between culture and globalization, and also to a limited extent the ties and differences, if any, between cultural liberty and happiness. This paper attempts to relate the concept of cultural liberty to the idea of Gross National Happiness. The author underlines strongly that the culture discussed is not about the particular culture of Bhutan; it is about culture in general and in abstract. Likewise happiness referred to here i...

  5. How culture affects management?

    OpenAIRE

    Billi, Lorena

    2012-01-01

    The study is about how culture affects management. Culture can have many different meanings. Management has also many different ways to be approached. While doing research about cultures, the study will try to analyze how the culture affects the management. The study starts with a full explanation of the meaning of culture. Some previous analysis and studies are added to illustrate my study on the subject. The effect culture has on management is studied at different levels. The study does not...

  6. Cultural Context and Translation

    Institute of Scientific and Technical Information of China (English)

    张敏

    2009-01-01

    cultural context plays an important role in translation. Because translation is a cross-culture activity, the culture context that influ-ences translating is consisted of both the culture contexts of source language and target language. This article firstly analyzes the concept of context and cultural context, then according to the procedure of translating classifies cultural context into two stages and talks about how they respectively influence translating.

  7. Cultural influences on personality.

    Science.gov (United States)

    Triandis, Harry C; Suh, Eunkook M

    2002-01-01

    Ecologies shape cultures; cultures influence the development of personalities. There are both universal and culture-specific aspects of variation in personality. Some culture-specific aspects correspond to cultural syndromes such as complexity, tightness, individualism, and collectivism. A large body of literature suggests that the Big Five personality factors emerge in various cultures. However, caution is required in arguing for such universality, because most studies have not included emic (culture-specific) traits and have not studied samples that are extremely different in culture from Western samples.

  8. Safeguards Culture: Analogies from Safety Culture and Security Culture

    International Nuclear Information System (INIS)

    Naito, K.

    2013-01-01

    The terminology of 'safeguards culture' has been used loosely by safeguards experts as an essential element for establishing an organizational environment of stakeholders for the effective and efficient implementation of international safeguards. However, unlike the other two triplet brothers/ sisters of 3S's (Safety, Security, Safeguards), there is no formally established definition of safeguards culture. In the case of safety culture, INSAG (the International Nuclear Safety Advisory Group) has extensively dealt with its concept, elaborating its definition and key characteristics, and published its report, INSAG-4, as the IAEA Safety Series 75. On the other hand, security culture has also been defined by AdSec (the Advisory Group on Nuclear Security). In this paper, a provisional definition of safeguards culture is made on the analogies of safety culture and security culture, and an effort is made to describe essential elements of safeguards culture. It is proposed for SAGSI (the Standing Advisory Group on Safeguards Implementation) to formally consider the definition of safeguards culture and its characteristics. The paper is followed by the slides of the presentation. (author)

  9. Cultural Analysis - towards cross-cultural understanding

    DEFF Research Database (Denmark)

    Gullestrup, Hans

    The book considers intercultural understanding and co-action, partly by means of general insight into concept of culture and the dimensions which bring about cultural differences, and partly as a methodology to analyse a certain culture - whether one's own or others'. This leads towards an unders......The book considers intercultural understanding and co-action, partly by means of general insight into concept of culture and the dimensions which bring about cultural differences, and partly as a methodology to analyse a certain culture - whether one's own or others'. This leads towards...... a theoretical/abstract proposal for cultural understanding. The second part presents a theoretical/abstract proposal for under-standing intercultural plurality and complexity. The third part provides an empirical model for the analysis of intercultural co-action. Finally, the fourth part present and discusses...

  10. FROM CULTURAL IMPOTENCE TO CULTURAL AMPUTATION

    Directory of Open Access Journals (Sweden)

    Вячеслав Владимирович Суханов

    2013-04-01

    Full Text Available Cultural space of any state is formed by a population that is within its borders. In this article, the author introduces a new cultural definitions «cultural impotence» and «cultural amputation», justifying their use, both in terms of population of the Russian Federation and the European Union and America. The article analyzes the state of society and the cultural factors that influence the development of society in Russia, there are options to bring the country out of a deep cultural crisis. Also established a close relationship between the domestic policy of the state and development of culture.DOI: http://dx.doi.org/10.12731/2218-7405-2013-2-1

  11. Space of solitude in culture

    Directory of Open Access Journals (Sweden)

    Agnieszka Małgorzata Kulig

    2014-04-01

    Full Text Available In my article I suggest looking at the phenomenon of solitude in culture not only as an opportunity to study an intimate and individual experience, to which numerous descriptions refer. I would rather consider whether possessing knowledge about this experience we acquire competencies regarding the culture in which this solitude has occurred. The discourse of melancholy, for which the space of solitude and confrontation with the ultimate has been the natural environment, supports me in describing the space of solitude in culture. For the purposes of reflection, I initiate my own definition of solitude as a space between life and death. That space allows the individual to feel the energy of life and the presence of death, even in such moments of human life (youth when the individual is not fully aware of their role in society, and is in the process of crystallizing their identity. In this case, solitude as a space between would be a state of suspension, which, however, prepares for active, independent life in the community. In this article, I refer to the postulate of the German culture researcher Thomas Macho in order to treat the experience of solitude as a context, an opportunity to practise techniques of culture, as well as to heterotopic space as defined by Michel Foucault.

  12. Enteral Feeding Set Handling Techniques.

    Science.gov (United States)

    Lyman, Beth; Williams, Maria; Sollazzo, Janet; Hayden, Ashley; Hensley, Pam; Dai, Hongying; Roberts, Cristine

    2017-04-01

    Enteral nutrition therapy is common practice in pediatric clinical settings. Often patients will receive a pump-assisted bolus feeding over 30 minutes several times per day using the same enteral feeding set (EFS). This study aims to determine the safest and most efficacious way to handle the EFS between feedings. Three EFS handling techniques were compared through simulation for bacterial growth, nursing time, and supply costs: (1) rinsing the EFS with sterile water after each feeding, (2) refrigerating the EFS between feedings, and (3) using a ready-to-hang (RTH) product maintained at room temperature. Cultures were obtained at baseline, hour 12, and hour 21 of the 24-hour cycle. A time-in-motion analysis was conducted and reported in average number of seconds to complete each procedure. Supply costs were inventoried for 1 month comparing the actual usage to our estimated usage. Of 1080 cultures obtained, the overall bacterial growth rate was 8.7%. The rinse and refrigeration techniques displayed similar bacterial growth (11.4% vs 10.3%, P = .63). The RTH technique displayed the least bacterial growth of any method (4.4%, P = .002). The time analysis in minutes showed the rinse method was the most time-consuming (44.8 ± 2.7) vs refrigeration (35.8 ± 2.6) and RTH (31.08 ± 0.6) ( P refrigerating the EFS between uses is the next most efficacious method for handling the EFS between bolus feeds.

  13. Laboratory techniques for human embryos.

    Science.gov (United States)

    Geber, Selmo; Sales, Liana; Sampaio, Marcos A C

    2002-01-01

    This review is concerned with laboratory techniques needed for assisted conception, particularly the handling of gametes and embryos. Such methods are being increasingly refined. Successive stages of fertilization and embryogenesis require especial care, and often involve the use of micromanipulative methods for intracytoplasmic sperm injection (ICSI) or preimplantation genetic diagnosis. Embryologists must take responsibility for gamete collection and preparation, and for deciding on the means of insemination or ICSI. Embryos must be assessed in culture, during the 1-cell, cleaving and morula/blastocyst stages, and classified according to quality. Co-culture methods may be necessary. The best embryos for transfer must be selected and loaded into the transfer catheter. Embryos not transferred must be cryopreserved, which demands the correct application of current methods of media preparation, seeding and the correct speed for cooling and warming. Before too long, methods of detecting abnormal embryos and avoiding their transfer may become widespread.

  14. Culture Elements in Intercultural Communication:Phenomena and Strategies%Culture Elements in Intercultural Communication: Phenomena and Strategies

    Institute of Scientific and Technical Information of China (English)

    刘永安

    2017-01-01

    With the advancement of globalization and the"the one-belt and one-road"initiative, there is greater-than-ever need for intercultural communication in many fields. With distinguished cultures, there will be conflicts of all kinds in intercultur-al communication, which greatly hinder the intercultural communication. The research to explore the culture elements and the cultural interference is of great significance for intercultural communication. Herein, culture elements and culture interference are to be explored, and strategies and techniques to minimize cultural interference are put forward, so as to promote intercultural communication.

  15. Socio-cultural inhibitors to use of modern contraceptive techniques ...

    African Journals Online (AJOL)

    Methods: this was a qualitative study conducted in 2012 among men aged 15-64 and ... All interviews were tape recorded, translated and transcribed verbatim. ... a latent content analysis approach, with support of ATLAS.ti qualitative software.

  16. Micropropagation of Eucalyptus grandis and nitens using tissue culture techniques

    Energy Technology Data Exchange (ETDEWEB)

    Furze, M.J.; Creswell, C.F.

    1985-01-01

    Experiments with nodal explants of E. grandis and E. nitens seedlings and E. grandis coppice shoots showed that a large number of plants can be produced from a single explant using micropropagation. The percentage of micropropagated shoots that formed roots was about 90% for E. grandis and 80% for E. nitens. For both species, about 90% of the rooted shoots survived after hardening off. 9 references.

  17. Propagation of pistachio applying in vitro culture techniques

    OpenAIRE

    Marín Velázquez, Juan Antonio; García Martín, Elena; Lorente Alonso, Pilar; Arbeloa Matute, Arancha; Andreu Puyal, Pilar

    2017-01-01

    8 Pags.- 2 Tabls.- 2 Figs. Article derived of a previous contribution of the authors to VI International Symposium on Production and Establishment of Micropropagated Plants (San Remo, Italy. April 2015). The defnitive version is available at: https://www.actahort.org/index.htm

  18. Misting cooling technique for protected culture of Oncidium orchids ...

    African Journals Online (AJOL)

    user

    2012-10-04

    Oct 4, 2012 ... 2Department of Bio-industrial Mechatronics Engineering, National Ch5ungHsing University, 250 Kuokuang Road,. Taichung, Taiwan. ... In this study, mechanical ventilation fans with intermittent misting were ... are in Taiwan and Malaysia. .... An 11.0 × 30 × 3.5 m high, 1155 m3 volume, plastic-glazed steel-.

  19. Many Forms of Culture

    Science.gov (United States)

    Cohen, Adam B.

    2009-01-01

    Psychologists interested in culture have focused primarily on East-West differences in individualism-collectivism, or independent-interdependent self-construal. As important as this dimension is, there are many other forms of culture with many dimensions of cultural variability. Selecting from among the many understudied cultures in psychology,…

  20. Reflections on Culture.

    Science.gov (United States)

    Fox, Lisette

    1999-01-01

    Examines aspects of cross-cultural studies, delineates a concept of culture, explores the interplay of culture and ethics, and analyzes the shifting cultural and economic values as issues that either the economists or the humanists will deal with under the banner of the future global village. (Author/VWL)