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Sample records for hypocotyl segment explants

  1. Increased Agrobacterium-mediated transformation and rooting efficiencies in canola (Brassica napus L.) from hypocotyl segment explants

    Science.gov (United States)

    Cardoza, V.; Stewart, C. N.

    2003-01-01

    An efficient protocol for the production of transgenic Brassica napus cv. Westar plants was developed by optimizing two important parameters: preconditioning time and co-cultivation time. Agrobacterium tumefaciens-mediated transformation was performed using hypocotyls as explant tissue. Two variants of a green fluorescent protein (GFP)-encoding gene--mGFP5-ER and eGFP--both under the constitutive expression of the cauliflower mosaic virus 35S promoter, were used for the experiments. Optimizing the preconditioning time to 72 h and co-cultivation time with Agrobacterium to 48 h provided the increase in the transformation efficiency from a baseline of 4% to 25%. With mGFP5-ER, the transformation rate was 17% and with eGFP it was 25%. Transgenic shoots were selected on 200 mg/l kanamycin. Rooting efficiency was 100% on half-strength Murashige and Skoog medium with 10 g/l sucrose and 0.5 mg/l indole butyric acid in the presence of kanamycin.

  2. Callus induction from epicotyl and hypocotyl explants of Parkia ...

    African Journals Online (AJOL)

    Epicotyl and hypocotyl explants of Parkia biglobosa (Locust bean) were cultured in vitro to investigate their callogenic capacity. Established cultures were obtained and maintained on MS medium supplemented with either 2,4-D or NAA, each of concentration range of 0.4 – 1.0 mg/L. In general, while higher concentrations of ...

  3. In vitro performances of hypocotyl and cotyledon explants of tomato ...

    African Journals Online (AJOL)

    A plant tissue culture technique is a good method for the evaluation and screening of plant genotypes for salt tolerance. In vitro evaluations of sodium chloride (NaCl) effects on two tomato cultivars (Pearl and Beril) were investigated with four NaCl levels (0, 25, 50 and 75 mM) using hypocotyl and cotyledon explants.

  4. In vitro response from cotyledon and hypocotyls explants in tomato ...

    African Journals Online (AJOL)

    USER

    2010-07-26

    Jul 26, 2010 ... 1Institute of Tropical Agriculture, University of Putra Malaysia, 43400 Serdang, Selangor, Malaysia. 2Faculty of Agriculture ... shoot length was significantly different between cotyledon and hypocotyls derived explants. No adventitious shoots ... growth regulator used in culture medium (Bhatia et al.,. 2004a).

  5. Direct adventitious shoot bud formation on hypocotyls explants in Millettia pinnata (L.) Panigrahi- a biodiesel producing medicinal tree species

    OpenAIRE

    Nagar, Durga Singh; Jha, Suman Kumar; Jani, Jigar

    2015-01-01

    A reproducible protocol developed for in vitro regeneration of Milletia pinnata using hypocotyl segments. Multiple shoots were induced from hypocotyl explants through direct adventitious shoot bud regeneration. The proximal end of hypocotyls was responsive for shoot bud induction. Silver nitrate and adenine sulphate had a positive effect on shoot bud induction and elongation. The maximum response and number of shoot bud produced in media supplemented with 8.88 μM BAP with 108.6 μM adenine sul...

  6. Direct adventitious shoot bud formation on hypocotyls explants in Millettia pinnata (L.) Panigrahi- a biodiesel producing medicinal tree species.

    Science.gov (United States)

    Nagar, Durga Singh; Jha, Suman Kumar; Jani, Jigar

    2015-04-01

    A reproducible protocol developed for in vitro regeneration of Milletia pinnata using hypocotyl segments. Multiple shoots were induced from hypocotyl explants through direct adventitious shoot bud regeneration. The proximal end of hypocotyls was responsive for shoot bud induction. Silver nitrate and adenine sulphate had a positive effect on shoot bud induction and elongation. The maximum response and number of shoot bud produced in media supplemented with 8.88 μM BAP with 108.6 μM adenine sulphate and 11.84 μM silver nitrate. Elongated shoots were harvested and successful rooting of microshoots achieved on MS media supplemented with 9.84 μM IBA, with 81.1 % rooting. Remaining shoot buds sub-cultured for further multiplication and elongation. Each subculture produced eight to nine elongated microshoots up to four subcultures. The rooted microshoots were successfully hardened and transferred to field.

  7. In vitro performances of hypocotyl and cotyledon explants of tomato ...

    African Journals Online (AJOL)

    Jane

    2011-08-15

    Aug 15, 2011 ... Key words: Cotyledons, hypocotyls, In vitro, salt stress, tomato. ... the use of irrigation water containing a high percentage of salts, in ... of small scale with clear visibility for observation shoot ..... Metabolic fingerprinting of salt.

  8. Effect of Hormones on Direct Shoot Regeneration in Hypocotyl Explants of Tomato

    Directory of Open Access Journals (Sweden)

    Rizwan RASHID

    2010-03-01

    Full Text Available This study was conducted for developing a high frequency regeneration system in two genotypes of tomato (Lycopersicon esculentum Mill., �Punjab Upma� and �IPA-3� for direct shoot regeneration from hypocotyl explants. The explants were excised from in vitro tomato seedlings and cultured on MS medium supplemented with different concentrations and combinations of hormones. Direct regeneration was significantly influenced by the genotype hormones combination and concentrations. The MS medium supplemented with (Kinetin 0.5 mg/l and (BAP 0.5 mg/l was found optimum for inducing direct shoot regeneration and number of shoots per explant from hypocotyl explants on this medium. Shoot regeneration per cent in �Punjab Upma� and �IPA-3� per cent was recorded to be highest i.e (86.02 and (82.57 respectively. Besides this, average number shoots per explant was also highest i.e (3.16 in case of �Punjab Upma� and (2.93 in case of �IPA-3�. A significant decline was observed in percent shoot regeneration and average number of shoots per explant with increase in the hormonal concentration. Shoots were obtained and transferred to the elongation medium (MS + BAP 0.3 mg/l. Hundred per cent rooting was induced in separated shoots upon culturing on MS and � MS basal media. Hardening on moist cotton showed maximum plantlet survival rate in case of both genotypes. After hardening, plants were transferred to soil. Thus, a tissue culture base line was established in tomato for obtaining direct regeneration using hypocotyl as explants.

  9. Effect of Hormones on Direct Shoot Regeneration in Hypocotyl Explants of Tomato

    Directory of Open Access Journals (Sweden)

    Rizwan RASHID

    2010-03-01

    Full Text Available This study was conducted for developing a high frequency regeneration system in two genotypes of tomato (Lycopersicon esculentum Mill., Punjab Upma and IPA-3 for direct shoot regeneration from hypocotyl explants. The explants were excised from in vitro tomato seedlings and cultured on MS medium supplemented with different concentrations and combinations of hormones. Direct regeneration was significantly influenced by the genotype hormones combination and concentrations. The MS medium supplemented with (Kinetin 0.5 mg/l and (BAP 0.5 mg/l was found optimum for inducing direct shoot regeneration and number of shoots per explant from hypocotyl explants on this medium. Shoot regeneration per cent in Punjab Upma and IPA-3 per cent was recorded to be highest i.e (86.02 and (82.57 respectively. Besides this, average number shoots per explant was also highest i.e (3.16 in case of Punjab Upma and (2.93 in case of IPA-3. A significant decline was observed in percent shoot regeneration and average number of shoots per explant with increase in the hormonal concentration. Shoots were obtained and transferred to the elongation medium (MS + BAP 0.3 mg/l. Hundred per cent rooting was induced in separated shoots upon culturing on MS and MS basal media. Hardening on moist cotton showed maximum plantlet survival rate in case of both genotypes. After hardening, plants were transferred to soil. Thus, a tissue culture base line was established in tomato for obtaining direct regeneration using hypocotyl as explants.

  10. Development of efficient catharanthus roseus regeneration and transformation system using agrobacterium tumefaciens and hypocotyls as explants

    Directory of Open Access Journals (Sweden)

    Wang Quan

    2012-06-01

    Full Text Available Abstract Background As a valuable medicinal plant, Madagascar periwinkle (Catharanthus roseus produces many terpenoid indole alkaloids (TIAs, such as vindoline, ajamlicine, serpentine, catharanthine, vinblastine and vincristine et al. Some of them are important components of drugs treating cancer and hypertension. However, the yields of these TIAs are low in wild-type plants, and the total chemical synthesis is impractical in large scale due to high-cost and their complicated structures. The recent development of metabolic engineering strategy offers a promising solution. In order to improve the production of TIAs in C. roseus, the establishment of an efficient genetic transformation method is required. Results To develop a genetic transformation method for C. roseus, Agrobacterium tumefaciens strain EHA105 was employed which harbors a binary vector pCAMBIA2301 containing a report β-glucuronidase (GUS gene and a selectable marker neomycin phosphotransferase II gene (NTPII. The influential factors were investigated systematically and the optimal transformation condition was achieved using hypocotyls as explants, including the sonication treatment of 10 min with 80 W, A. tumefaciens infection of 30 min and co-cultivation of 2 d in 1/2 MS medium containing 100 μM acetosyringone. With a series of selection in callus, shoot and root inducing kanamycin-containing resistance media, we successfully obtained stable transgenic regeneration plants. The expression of GUS gene was confirmed by histochemistry, polymerase chain reaction, and genomic southern blot analysis. To prove the efficiency of the established genetic transformation system, the rate-limiting gene in TIAs biosynthetic pathway, DAT, which encodes deacetylvindoline-4-O-acetyltransferase, was transferred into C. roseus using this established system and 9 independent transgenic plants were obtained. The results of metabolite analysis using high performance liquid chromatography (HPLC

  11. High frequency organogenesis in hypocotyl, cotyledon, leaf and petiole explants of broccoli (Brassica oleracea L. var. italica), an important vegetable crop.

    Science.gov (United States)

    Kumar, Pankaj; Srivastava, D K

    2015-04-01

    Broccoli (Brassica oleracea L. var. italica) is an important, nutritionally rich vegetable crop, but severely affected by environmental stresses, pests and diseases which cause massive yield and quality losses. Genetic manipulation is becoming an important method for broccoli improvement. In the present study, a reproducible and highly efficient protocol for obtaining organogenesis from hypocotyl, cotyledon, leaf and petiole explants of broccoli (Brassica oleracea L. var. italica cv. Solan green head) has been developed. Hypocotyl and cotyledon explants were used from 10 to 12 days old aseptically grown seedlings whereas leaf and petiole explants were excised from 18 to 20 days old green house grown seedlings and surface sterilized. These explants were cultured on shoot induction medium containing different concentration and combination of BAP and NAA. High efficiency shoot regeneration has been achieved in hypocotyl (83.33 %), cotyledon (90.11 %), leaf (62.96 %) and petiole (91.10 %) explants on MS medium supplemented with 3.5 mg/l BAP + 0.019 mg/l NAA 2.5 mg/l BAP + 0.5 mg/l NAA, 4.0 mg/l BAP + 0.5 mg/l NAA and 4.5 mg/l BAP + 0.019 mg/l NAA respectively. Petiole explants showed maximum shoot regeneration response as compared to other explants. MS medium supplemented with 0.10 mg/l NAA was found best for root regeneration (100 %) from in vitro developed shoots. The regenerated complete plantlets were transferred to the pots containing cocopeat and successfully acclimatized. This optimized regeneration protocol can be efficiently used for genetic transformation in broccoli. This is the first comparative report on multiple shoot induction using four different types of explants viz. hypocotyl, cotyledon, leaf and petiole.

  12. Agrobacterium-mediated genetic transformation of Fraxinus americana hypocotyls

    Science.gov (United States)

    Kaitlin J. Palla; Paula M. Pijut

    2015-01-01

    An Agrobacterium tumefaciens-mediated genetic transformation system was successfully developed for white ash (Fraxinus americana) using hypocotyls as the initial explants. Hypocotyls isolated from mature embryos germinated on Murashige and Skoog (MS) medium supplemented with 22.2 µM 6-benzyladenine (BA) and 0.5 µM...

  13. Differential Effect of Auxin on Molecular Weight Distributions of Xyloglucans in Cell Walls of Outer and Inner Tissues from Segments of Dark Grown Squash (Cucurbita maxima Duch.) Hypocotyls.

    Science.gov (United States)

    Wakabayashi, K; Sakurai, N; Kuraishi, S

    1991-04-01

    Effects of indole-3-acetic acid (IAA) on the mechanical properties of cell walls and structures of cell wall polysaccharides in outer and inner tissues of segments of dark grown squash (Cucurbita maxima Duch.) hypocotyls were investigated. IAA induced the elongation of unpeeled, intact segments, but had no effect on the elongation of peeled segments. IAA induced the cell wall loosening in outer tissues as studied by the stress-relaxation analysis but not in inner tissues. IAA-induced changes in the net sugar content of cell wall fractions in outer and inner tissues were very small. Extracted hemicellulosic xyloglucans derived from outer tissues had a molecular weight about two times as large as in inner tissues, and the molecular weight of xyloglucans in both outer and inner tissues decreased during incubation. IAA substantially accelerated the depolymerization of xyloglucans in outer tissues, while it prevented that in inner tissues. These results suggest that IAA-induced growth in intact segments is due to the cell wall loosening in outer tissues, and that IAA-accelerated depolymerization of hemicellulosic xyloglucans in outer tissues is involved in the cell wall loosening processes.

  14. Sugar Composition and Molecular Weight Distribution of Cell Wall Polysaccharides in Outer and Inner Tissues from Segments of Dark Grown Squash (Cucurbita maxima Duch.) Hypocotyls.

    Science.gov (United States)

    Wakabayashi, K; Sakurai, N; Kuraishi, S

    1990-07-01

    The elongation growth of stem segments is determined by the outer cell layers (epidermis and collenchyma). We measured the sugar composition and molecular weight distribution of pectin and hemicellulose fractions obtained from inner and outer tissues of squash (Cucurbita maxima Duch.) hypocotyls. In addition, we studied the changes in these parameters after a 9 hour period of incubation of the segments. The results show that outer tissues have higher molecular weight pectin and hemicellulose compared to inner tissues (2-3 times higher). Incubation results in a 13 to 25% decrease in the amount of pectin and hemicellulose in inner tissues and an increase of 11 to 32% in the outer tissues. This increase in the outer tissues is accompanied by a decrease in the molecular weight of some of the components. These results clearly show that cell wall metabolism during elongation growth differs markedly in inner and outer tissues, and that future studies on the effect of auxin need to take these differences into account.

  15. Sugar Composition and Molecular Weight Distribution of Cell Wall Polysaccharides in Outer and Inner Tissues from Segments of Dark Grown Squash (Cucurbita maxima Duch.) Hypocotyls 1

    Science.gov (United States)

    Wakabayashi, Kazuyuki; Sakurai, Naoki; Kuraishi, Susumu

    1990-01-01

    The elongation growth of stem segments is determined by the outer cell layers (epidermis and collenchyma). We measured the sugar composition and molecular weight distribution of pectin and hemicellulose fractions obtained from inner and outer tissues of squash (Cucurbita maxima Duch.) hypocotyls. In addition, we studied the changes in these parameters after a 9 hour period of incubation of the segments. The results show that outer tissues have higher molecular weight pectin and hemicellulose compared to inner tissues (2-3 times higher). Incubation results in a 13 to 25% decrease in the amount of pectin and hemicellulose in inner tissues and an increase of 11 to 32% in the outer tissues. This increase in the outer tissues is accompanied by a decrease in the molecular weight of some of the components. These results clearly show that cell wall metabolism during elongation growth differs markedly in inner and outer tissues, and that future studies on the effect of auxin need to take these differences into account. PMID:16667612

  16. Agrobacterium-mediated transformation of Fraxinus pennsylvanica hypocotyls and plant regeneration

    Science.gov (United States)

    Ningxia Du; Paula M. Pijut

    2009-01-01

    A genetic transformation protocol for green ash (Fraxinus pennsylvanica) hypocotyl explants was developed. Green ash hypocotyls were transformed using Agrobacterium tumefaciens strain EHA105 harboring binary vector pq35GR containing the neomycin phosphotransferase (nptII) and β-glucuronidase (GUS) fusion...

  17. Interactions of phytochromes A, B1 and B2 in light-induced competence for adventitious shoot formation in hypocotyl of tomato (Solanum lycopersicum L.).

    Science.gov (United States)

    Lercari, B; Bertram, L

    2004-02-01

    The interactions of phytochrome A (phyA), phytochrome B1 (phyB1) and phytochrome B2 (phyB2) in light-dependent shoot regeneration from the hypocotyl of tomato was analysed using all eight possible homozygous allelic combinations of the null mutants. The donor plants were pre-grown either in the dark or under red or far-red light for 8 days after sowing; thereafter hypocotyl segments (apical, middle and basal portions) were transferred onto hormone-free medium for culture under different light qualities. Etiolated apical segments cultured in vitro under white light showed a very high frequency of regeneration for all of the genotypes tested besides phyB1phyB2, phyAphyB1 and phyAphyB1phyB2 mutants. Evidence is provided of a specific interference of phyB2 with phyA-mediated HIR to far-red and blue light in etiolated explants. Pre-treatment of donor plants by growth under red light enhanced the competence of phyB1phyB2, phyAphyB1 and phyAphyB1phyB2 mutants for shoot regeneration, whereas pre-irradiation with far-red light enhanced the frequency of regeneration only in the phyAphyB1 mutant. Multiple phytochromes are involved in red light- and far-red light-dependent acquisition of competence for shoot regeneration. The position of the segments along the hypocotyl influenced the role of the various phytochromes and the interactions between them. The culture of competent hypocotyl segments under red, far-red or blue light reduced the frequency of explants forming shoots compared to those cultured under white light, with different genotypes having different response patterns.

  18. Induction of hairy roots by various strains of Agrobacterium rhizogenes in different types of Capsicum species explants.

    Science.gov (United States)

    Md Setamam, Nursuria; Jaafar Sidik, Norrizah; Abdul Rahman, Zainon; Che Mohd Zain, Che Radziah

    2014-06-30

    Capsicum annuum and Capsicum frutescens, also known as "chilies", belong to the Solanaceae family and have tremendous beneficial properties. The application of hairy root culture may become an alternative method for future development of these species by adding value, such as by increasing secondary metabolites and improving genetic and biochemical stability compared with normal Capsicum plants. Therefore, in this research, different types of explants of both species were infected with various Agrobacterium rhizogenes strains to provide more information about the morphology and induction efficiency of hairy roots. After 2 weeks of in vitro seed germination, young seedling explants were cut into three segments; the cotyledon, hypocotyl, and radical. Then, the explants were co-cultured with four isolated A. rhizogenes strains in Murashige & Skoog culture media (MS) containing decreasing carbenicillin disodium concentrations for one month. In this experiment, thick and short hairy roots were induced at all induction sites of C. annuum while thin, elongated hairy roots appeared mostly at wound sites of C. frutescens. Overall, the hairy root induction percentages of C. frutescens were higher compared with C. annuum. Hairy root initiation was observed earliest using radicles (1st week), followed by cotyledons (2nd week), and hypocotyls (3rd week). Cotyledon explants of both species had the highest induction frequency with all strains compared with the other explants types. Strains ATCC 13333 and ATCC 15834 were the most favourable for C. frutescens while ATCC 43056 and ATCC 43057 were the most favourable for C. annuum. The interactions between the different explants and strains showed significant differences with p-values Capsicum species. Both Capsicum species were amenable to A. rhizogenes infection and hairy root induction is recommended for use as an alternative explants in future plant-based studies.

  19. Differential Effect of Auxin on Molecular Weight Distributions of Xyloglucans in Cell Walls of Outer and Inner Tissues from Segments of Dark Grown Squash (Cucurbita maxima Duch.) Hypocotyls 1

    Science.gov (United States)

    Wakabayashi, Kazuyuki; Sakurai, Naoki; Kuraishi, Susumu

    1991-01-01

    Effects of indole-3-acetic acid (IAA) on the mechanical properties of cell walls and structures of cell wall polysaccharides in outer and inner tissues of segments of dark grown squash (Cucurbita maxima Duch.) hypocotyls were investigated. IAA induced the elongation of unpeeled, intact segments, but had no effect on the elongation of peeled segments. IAA induced the cell wall loosening in outer tissues as studied by the stress-relaxation analysis but not in inner tissues. IAA-induced changes in the net sugar content of cell wall fractions in outer and inner tissues were very small. Extracted hemicellulosic xyloglucans derived from outer tissues had a molecular weight about two times as large as in inner tissues, and the molecular weight of xyloglucans in both outer and inner tissues decreased during incubation. IAA substantially accelerated the depolymerization of xyloglucans in outer tissues, while it prevented that in inner tissues. These results suggest that IAA-induced growth in intact segments is due to the cell wall loosening in outer tissues, and that IAA-accelerated depolymerization of hemicellulosic xyloglucans in outer tissues is involved in the cell wall loosening processes. PMID:16668092

  20. callus induction from epicotyl and hypocotyl explants of

    African Journals Online (AJOL)

    AMO0+ and B.E. AYISIRE. Department of Botany, Obafemi Awolowo University, Ile-Ife. Nigeria. (Submitted: 31 May 2004; Accepted: 31 October 2004). Abstract ..... of Nigeria: Implications for food security. pp. 265-. 283. University of Agriculture, Abeokuta, Conference. Proceedings Series No. 3. Ayisire, B.E., Obembe, 0.0.

  1. In vitro response from cotyledon and hypocotyls explants in tomato ...

    African Journals Online (AJOL)

    USER

    2010-07-26

    Jul 26, 2010 ... randomized design (CRD) with four replications. .... culture, transformation and molecular markers for crops improvement. Science ... applications. Trends Genet. ... Culture Concepts and Laboratory Exercises. New York, USA: ...

  2. Flask sealing on in vitro seed germination and morphogenesis of two types of ornamental pepper explants

    Directory of Open Access Journals (Sweden)

    Diego Silva Batista

    Full Text Available ABSTRACT: The influence of flask sealing and explant source on the in vitro morphogenesis of pepper, were evaluated in Capsicum annuum. Seeds were sterilized and inoculated in Murashige and Skoog media supplemented with vitamins, myo-inositol, sucrose, and agar. Gas exchange was evaluated in the germination stage by comparing 3 flask-sealing systems: rigid polypropylene lids (PLs without vents, PLs with 1 vent, and PLs with 2 vents covered with membranes. In the regeneration stage, cotyledon and hypocotyl segments were transferred to the organogenesis-inducing media, being the different sealing types also tested in a factorial scheme. Photosynthetic pigments, morphological and hystological analyses were conducted for each treatment. Plants maintained in glass flasks capped with vented lids showed more vigorous growth and differentiated anatomical structures. These treatments resulted in taller plants, higher numbers and more expanded leaves, higher fresh and dry weights, and an increase in photosynthetic pigments. Cultivation of C. annuum in flasks with reduced gas exchange was more effective on callus induction. During the regeneration stage, hypocotyls were more effective than cotyledons. Sealing type influenced the morphogenic responses of pepper, demonstrating that an increase in gas exchange has a positive effect on biomass production and acclimatization of the plantlets.

  3. Plantlet regeneration potential from seedling explants of vitegnus (Vitex agnus castus).

    Science.gov (United States)

    Chamandoosti, F

    2007-11-15

    In this research a simple and repeatable method for regeneration of a important medicinal plant (Vitex agnus castus) described. Different seedling explants such as hypocotyl, cotyledon, root and apical meristem were cultured in MS basal media with different kinds and concentrations of PGRs. Root and apical meristem explants were the only explants that have regeneration whole plantlets potential. It was interesting that regeneration whole plantlets from root and apical meristem explants have different developmental pathways. Whole plantlets from apical meristem explants regenerated by passing phase callusing whereas regeneration whole plantlets from root was direct and without phase callusing. This subject implies that we can have many manipulation possibilities in order to different objects of tissue culture by selecting different explants in vitegnus.

  4. Development of efficient plant regeneration and transformation system for impatiens using Agrobacterium tumefaciens and multiple bud cultures as explants.

    Science.gov (United States)

    Dan, Yinghui; Baxter, Aaron; Zhang, Song; Pantazis, Christopher J; Veilleux, Richard E

    2010-08-09

    Impatiens (Impatiens walleriana) is a top selling floriculture crop. The potential for genetic transformation of Impatiens to introduce novel flower colors or virus resistance has been limited by its general recalcitrance to tissue culture and transformation manipulations. We have established a regeneration and transformation system for Impatiens that provides new alternatives to genetic improvement of this crop. In a first step towards the development of transgenic INSV-resistant Impatiens, we developed an efficient plant regeneration system using hypocotyl segments containing cotyledonary nodes as explants. With this regeneration system, 80% of explants produced an average of 32.3 elongated shoots per initial explant plated, with up to 167 elongated shoots produced per explant. Rooting efficiency was high, and 100% of shoots produced roots within 12 days under optimal conditions, allowing plant regeneration within approximately 8 weeks. Using this regeneration system, we developed an efficient Agrobacterium-mediated Impatiens transformation method using in vitro multiple bud cultures as explants and a binary plasmid (pHB2892) bearing gfp and nptII genes. Transgenic Impatiens plants, with a frequency up to 58.9%, were obtained within 12 to 16 weeks from inoculation to transfer of transgenic plants to soil. Transgenic plants were confirmed by Southern blot, phenotypic assays and T1 segregation analysis. Transgene expression was observed in leaves, stems, roots, flowers, and fruit. The transgenic plants were fertile and phenotypically normal. We report the development of a simple and efficient Agrobacterium-mediated transformation system for Impatiens. To the best of our knowledge, there have been no reports of Agrobacterium-mediated transformation of Impatiens with experimental evidence of stable integration of T-DNA and of Agrobacterium-mediated transformation method for plants using in vitro maintained multiple bud cultures as explants. This transformation system

  5. Development of Efficient Plant Regeneration and Transformation System for Impatiens Using Agrobacterium tumefaciens and Multiple Bud Cultures as Explants

    Directory of Open Access Journals (Sweden)

    Dan Yinghui

    2010-08-01

    Full Text Available Abstract Background Impatiens (Impatiens walleriana is a top selling floriculture crop. The potential for genetic transformation of Impatiens to introduce novel flower colors or virus resistance has been limited by its general recalcitrance to tissue culture and transformation manipulations. We have established a regeneration and transformation system for Impatiens that provides new alternatives to genetic improvement of this crop. Results In a first step towards the development of transgenic INSV-resistant Impatiens, we developed an efficient plant regeneration system using hypocotyl segments containing cotyledonary nodes as explants. With this regeneration system, 80% of explants produced an average of 32.3 elongated shoots per initial explant plated, with up to 167 elongated shoots produced per explant. Rooting efficiency was high, and 100% of shoots produced roots within 12 days under optimal conditions, allowing plant regeneration within approximately 8 weeks. Using this regeneration system, we developed an efficient Agrobacterium-mediated Impatiens transformation method using in vitro multiple bud cultures as explants and a binary plasmid (pHB2892 bearing gfp and nptII genes. Transgenic Impatiens plants, with a frequency up to 58.9%, were obtained within 12 to 16 weeks from inoculation to transfer of transgenic plants to soil. Transgenic plants were confirmed by Southern blot, phenotypic assays and T1 segregation analysis. Transgene expression was observed in leaves, stems, roots, flowers, and fruit. The transgenic plants were fertile and phenotypically normal. Conclusion We report the development of a simple and efficient Agrobacterium-mediated transformation system for Impatiens. To the best of our knowledge, there have been no reports of Agrobacterium-mediated transformation of Impatiens with experimental evidence of stable integration of T-DNA and of Agrobacterium-mediated transformation method for plants using in vitro maintained

  6. Effect of Pre-culture Irradiation and Explant Types on Efficiency of Brassica napus Genetic Transformation

    International Nuclear Information System (INIS)

    Amer, I.M.; Moustafa, H.A.M.; Azzam, C.R.

    2008-01-01

    The irradiated seeds of canola cv. Drakkar ( Brassica napus l. ) were germinated under aspect conditions, cotyledonary petioles and hypocotyl of 6 days old seedlings were used for Agrobacterium-mediated transformation. Agrobacterium tumefaciens has construct with the selectable marker gene (NPT II) and the desirable gene (HPPD). Direct and indirect shoot organogenesis were obtained from the both explants. Cotyledonary petioles was higher responded than hypocotyl with respective 26% and 14% of the explants producing NPT II-positive shoots after the selection on 50mg/l kanamycin. Calli might develop on and not in the agar medium were un transformation. This explains the higher number of escapes detected in hypocotyl explants than in experiments with cotyledons. The frequency of transformation plants as a function of indirect organogenesis was more than direct shoot regeneration from explants. The pre- irradiation with 75 Gy of gamma rays enhanced the genetic transformation frequencies by about 10 % as compared to that of the un-irradiated material. The obtained shoots were rooted and regenerated mature plants

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

    Directory of Open Access Journals (Sweden)

    L. Koohi,

    2014-07-01

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

  8. Protocols for Callus and Somatic Embryo Initiation for Hibiscus sabdariffa L. (Malvaceae): Influence of Explant Type, Sugar, and Plant Growth Regulators

    Science.gov (United States)

    A significant work on callus induction and somatic embryogenesis was realized for Hibiscus sabdariffa. Two genotypes (Hibiscus sabdariffa and Hibiscus sabdariffa var. altissima) two sugars (sucrose and glucose) and three concentrations (1 %, 2%, 3%) of each sugar, 3 explant types (root, hypocotyl, c...

  9. Regeneração in vitro de urucum (Bixa orellana L. a partir de diferentes tipos de explantes In vitro regeneration of annatto (Bixa orellana L. from various explants

    Directory of Open Access Journals (Sweden)

    Jane Fiuza Rodrigues Portela de Carvalho

    2005-12-01

    Full Text Available Este trabalho teve como objetivo avaliar a regeneração in vitro de plantas de urucum (Bixa orellana L. a partir de diferentes tipos de explantes. Para definir o meio de cultura adequado para indução de brotações, diferentes concentrações e, ou, combinações da auxina AIA e das citocininas BAP e ZEA foram testadas. As melhores respostas de regeneração para segmentos de hipocótilo, nós cotiledonares e hipocótilos invertidos foram observadas em meios suplementados de ZEA (2,28 µM e AIA (0,30 µM, ZEA (4,56 µM e ZEA (4,56 µM, respectivamente. O meio de enraizamento mais eficaz foi o MS, com a metade de sua concentração salina e 5 µM de AIB. Análises citológicas, realizadas antes da aclimatação, confirmaram a estabilidade cromossômica das plantas cultivadas in vitro, não sendo detectado variação com relação ao número de cromossomos metafásicos (2n = 14.The present work aimed the establishment of a regeneration protocol for annatto (Bixa orellana L. from different juvenile explants. In order to promote shoot induction, different concentrations and/or combinations of IAA and the cytokinins BAP and ZEA were assessed. Better regeneration responses were achieved when segmented hypocotyl, cotiledonary nodes and inverted hypocotyl were cultured onto MS-based medium supplemented with ZEA (2.28 µM and IAA (0.30 µM, ZEA (4.56 µM or ZEA (4.56 µM, respectively. Rooting of elongated shoots displayed higher frequencies when half-strength MS medium with IBA (5 µM was used. No genetic variation was detected among regenerants as revealed by cytological analysis based on metaphasic chromosome countings (2n = 14.

  10. In vitro plant regeneration of two cucumber (Cucumis sativum L. genotypes: Effects of explant types and culture medium

    Directory of Open Access Journals (Sweden)

    Grozeva Stanislava

    2014-01-01

    Full Text Available The effect of different phytohormone concentrations on callusogenesis and organogenesis in two cucumber genotypes were studied. It was established that the rate of plant regeneration depends on genotype, explant type and culture medium. Hypocotyls were found to be more responsive than cotyledons in morphogenesis. In vitro planlet-regenerants have been obtained in hypocotyls explants on culture medium with 1.0 and 2.0 mgL-1 BA for cultivar Gergana and in 1.0 and 3.0 mgL-1K-line 15B. Induction of regeneration in cotyledons were established only in cultivar Gergana on culture medium supplemented with 3.0 mgL-1 BA and in combination of 0.5 mgL-1IAA.

  11. System-wide organization of actin cytoskeleton determines organelle transport in hypocotyl plant cells

    Science.gov (United States)

    Nowak, Jacqueline; Ivakov, Alexander; Somssich, Marc; Persson, Staffan; Nikoloski, Zoran

    2017-01-01

    The actin cytoskeleton is an essential intracellular filamentous structure that underpins cellular transport and cytoplasmic streaming in plant cells. However, the system-level properties of actin-based cellular trafficking remain tenuous, largely due to the inability to quantify key features of the actin cytoskeleton. Here, we developed an automated image-based, network-driven framework to accurately segment and quantify actin cytoskeletal structures and Golgi transport. We show that the actin cytoskeleton in both growing and elongated hypocotyl cells has structural properties facilitating efficient transport. Our findings suggest that the erratic movement of Golgi is a stable cellular phenomenon that might optimize distribution efficiency of cell material. Moreover, we demonstrate that Golgi transport in hypocotyl cells can be accurately predicted from the actin network topology alone. Thus, our framework provides quantitative evidence for system-wide coordination of cellular transport in plant cells and can be readily applied to investigate cytoskeletal organization and transport in other organisms. PMID:28655850

  12. Effect Of Gamma Rays And Growth Regulators On Explants Excised From In Vitro Shoots And Greenhouse Seedlings, Of Pepper (Capsicum Annum L.)

    International Nuclear Information System (INIS)

    Maarouf, A. A.; Kassem, M.

    2004-01-01

    This experiment was conducted on pepper (Capsicum annum L.) to compare the ability of the in vitro explants with those of greenhouse grown seedlings on shoot proliferation and callus formation and their ability to form plantlets and the effect of gamma irradiation and growth regulators on the shoot tip, hypocotyls and leaf tissue was used as laboratory explants, leaf tissue nodes and internodes were taken from greenhouse seedlings. 6- benzyla-minopurine (BAP) in different concentrations was combined with Indoleacertic acid (IAA) to know their effect on shoot proliferation, 2,4 - Dichlorophenoxy acetic acid (2,4- D) was used for callus formation, and use stimulation effect of gamma irradiation, potassium nitrat (KNO 3 ), Thidaiazurom (TDZ) and casine hydrolysate (CH) for plantlet formation. The results showed that the highest percentage of callus was obtained by in vitro hypocotyls and greenhouse grown nodes followed by in vitro leaf tissue thereafter greenhouse leaf tissue. The shoot tips were the lowest efficient explants in producing callus in both in vitro and greenhouse ones. The highest percentage of shooting resulted from shoot tip, hypocotyls and leaf tissue of in vitro explants, followed by shoot tip, nodes and internodes of greenhouse grown explants and the lowest percentage was recorded by leaf tissue. Highest percentage of shoot number was obtained form greenhouse grown shoot tip followed by in vitro shoot tip, hypocotyls and leaf tissue of greenhouse grown seedlings the internodes were the lowest efficient in producing shoots. The highest success in plantlet formation was caused by TDZ followed by gamma irradiation and the other treatments were equaled. (Authors)

  13. Hypocotyl adventitious root organogenesis differs from lateral root development.

    Science.gov (United States)

    Verstraeten, Inge; Schotte, Sébastien; Geelen, Danny

    2014-01-01

    Wound-induced adventitious root (AR) formation is a requirement for plant survival upon root damage inflicted by pathogen attack, but also during the regeneration of plant stem cuttings for clonal propagation of elite plant varieties. Yet, adventitious rooting also takes place without wounding. This happens for example in etiolated Arabidopsis thaliana hypocotyls, in which AR initiate upon de-etiolation or in tomato seedlings, in which AR initiate upon flooding or high water availability. In the hypocotyl AR originate from a cell layer reminiscent to the pericycle in the primary root (PR) and the initiated AR share histological and developmental characteristics with lateral roots (LRs). In contrast to the PR however, the hypocotyl is a determinate structure with an established final number of cells. This points to differences between the induction of hypocotyl AR and LR on the PR, as the latter grows indeterminately. The induction of AR on the hypocotyl takes place in environmental conditions that differ from those that control LR formation. Hence, AR formation depends on differentially regulated gene products. Similarly to AR induction in stem cuttings, the capacity to induce hypocotyl AR is genotype-dependent and the plant growth regulator auxin is a key regulator controlling the rooting response. The hormones cytokinins, ethylene, jasmonic acid, and strigolactones in general reduce the root-inducing capacity. The involvement of this many regulators indicates that a tight control and fine-tuning of the initiation and emergence of AR exists. Recently, several genetic factors, specific to hypocotyl adventitious rooting in A. thaliana, have been uncovered. These factors reveal a dedicated signaling network that drives AR formation in the Arabidopsis hypocotyl. Here we provide an overview of the environmental and genetic factors controlling hypocotyl-born AR and we summarize how AR formation and the regulating factors of this organogenesis are distinct from LR

  14. Hypocotyl adventitious root organogenesis differs from lateral root development

    Directory of Open Access Journals (Sweden)

    Inge eVerstraeten

    2014-09-01

    Full Text Available Wound-induced adventitious root (AR formation is a requirement for plant survival upon root damage inflicted by pathogen attack, but also during the regeneration of plant stem cuttings for clonal propagation of elite plant varieties. Yet, adventitious rooting also takes place without wounding. This happens for example in etiolated Arabidopsis thaliana hypocotyls, in which AR initiate upon de-etiolation or in tomato seedlings, in which AR initiate upon flooding or high water availability. In the hypocotyl AR originate from a cell layer reminiscent to the pericycle in the primary root (PR and the initiated AR share histological and developmental characteristics with lateral roots (LR. In contrast to the PR however, the hypocotyl is a determinate structure with an established final number of cells. This points to differences between the induction of hypocotyl AR and LR on the PR, as the latter grows indeterminately. The induction of AR on the hypocotyl takes place in environmental conditions that differ from those that control LR formation. Hence, AR formation depends on differentially regulated gene products. Similarly to AR induction in stem cuttings, the capacity to induce hypocotyl AR is genotype-dependent and the plant growth regulator auxin is a key regulator controlling the rooting response. The hormones cytokinins, ethylene, jasmonic acid and strigolactones in general reduce the root-inducing capacity. The involvement of this many regulators indicates that a tight control and fine-tuning of the initiation and emergence of AR exists. Recently, several genetic factors, specific to hypocotyl adventitious rooting in Arabidopsis thaliana, have been uncovered. These factors reveal a dedicated signaling network that drives AR formation in the Arabidopsis hypocotyl. Here we provide an overview of the environmental and genetic factors controlling hypocotyl-born AR and we summarize how AR formation and the regulating factors of this organogenesis are

  15. A survey of the effect of explants type, plant growth regulators and activated charcoal on callus induction in Papaver bracteatum

    Directory of Open Access Journals (Sweden)

    Bahman Hosseini

    2015-09-01

    Full Text Available Callus culture is necessary for production of suspension cell culture in plant breeding programs. Regarding to the application of Papaver bracteatum as an important medicinal plant in production of benzophenantridine alkaloids, this study was performed to find the most suitable hormone combination and explant type for achieving to high percentage of callus induction fresh weight and somatic embryogenesis in this plant. For this purpose, hypocotyl explants were cultured in ½MS media containing active charcoal (2 and 4 mgL-1 in combination of different concentrations of NAA, 2,4-D (0, 1, 2, 3 and 5 mgL-1 and BA (0, 0.1 and 0.5 mgL-1. The seed explants were cultured in same treatments without active charcoal. Also, somatic embryogesis induction using seed explants in ½MS media containing different concentrations of NAA and 2,4-D (0, 0.5, 1 and 2 mgL-1 with BA 0.5 mgL-1 were investigated. The results showed that the highest percentage of callus induction (43.6%, 54% in hypocotyls explants were obtained in the ½MS media containing 2 mgL-1 active charcoal and 2 mgL-1 2,4-D and 5 mgL-1 NAA in companion with BA 0.5 mgL-1 respectively. The maximum callus induction (84% was obtained in ½MS medium with 1 mgL-1 2,4-D without active charcoal. The highest callus fresh weight (0.35% was obtained in MS media with 0.5 mgL-1 2,4-D andthe maximum rate of somatic embryogenesis induction (77% was observed in ½MS media containing 1 mgL-1 2,4-D with 0.5 mgL-1 BA.

  16. Effect of gamma-radiation on functioning of bean hypocotyl mitochondria: lipids and lipid-dependent enzymes of the electron transfer chain (ETC)

    Energy Technology Data Exchange (ETDEWEB)

    Pai, K U; Gaur, B K [Bhabha Atomic Research Centre, Bombay (India). Biology and Agriculture Div.

    1982-05-01

    A brief note presents the results of a study of the effect of ..gamma..-radiation on NADH-cytochrome c-reductase and succinate-cytochrome c-reductase of mitochondria from bean hypocotyl segments. About 2.5 cm long hypocotyl segments of 5-day-old kidney bean plants were exposed to 250 and 500 kR /sup 60/Co ..gamma..-rays at an exposure rate of 10 kR per min., maintaining the segments at 0 - 5/sup 0/C during irradiation. The results suggest that radiation adversely affects phospholipids, thereby lowering the activity of the dependent ETC enzymes in mitochondria. The results also indicate a possible radiation-induced destruction of the lipid moiety of co-enzyme Q/sub 10/.

  17. Cytoarchitecture in cultured rat neocortex explants

    NARCIS (Netherlands)

    de Jong, B. M.; Ruijter, J. M.; Romijn, H. J.

    1988-01-01

    Neocortex explants obtained from 6-day-old rat pups and cultured in a serum-free medium from 5 hr to 13 days in vitro (DIV) show preservation of cytoarchitectural characteristics. Major changes in the size of the explants and their layers occur during the first 2 DIV. A radial arrangement of neurons

  18. Proteomic analysis of soybean hypocotyl during recovery after flooding stress.

    Science.gov (United States)

    Khan, Mudassar Nawaz; Sakata, Katsumi; Komatsu, Setsuko

    2015-05-21

    Soybean is a nutritionally important crop, but exhibits reduced growth and yields under flooding stress. To investigate soybean responses during post-flooding recovery, a gel-free proteomic technique was used to examine the protein profile in the hypocotyl. Two-day-old soybeans were flooded for 2 days and hypocotyl was collected under flooding and during the post-flooding recovery period. A total of 498 and 70 proteins were significantly changed in control and post-flooding recovering soybeans, respectively. Based on proteomic and clustering analyses, three proteins were selected for mRNA expression and enzyme activity assays. Pyruvate kinase was increased under flooding, but gradually decreased during post-flooding recovery period at protein abundance, mRNA, and enzyme activity levels. Nucleotidylyl transferase was decreased under flooding and increased during post-flooding recovery at both mRNA expression and enzyme activity levels. Beta-ketoacyl reductase 1 was increased under flooding and decreased during recovery at protein abundance and mRNA expression levels, but its enzyme activity gradually increased during the post-flooding recovery period. These results suggest that pyruvate kinase, nucleotidylyl transferase, and beta-ketoacyl reductase play key roles in post-flooding recovery in soybean hypocotyl by promoting glycolysis for the generation of ATP and regulation of secondary metabolic pathways. This study analyzed post-flooding recovery response mechanisms in soybean hypocotyl, which is a model organ for studying secondary growth, using a gel-free proteomic technique. Mass spectrometry analysis of proteins extracted from soybean hypocotyls identified 20 common proteins between control and flooding-stressed soybeans that changed significantly in abundance over time. The hypocotyl proteins that changed during post-flooding recovery were assigned to protein, development, secondary metabolism, and glycolysis categories. The analysis revealed that three

  19. Lichen explants and natural occurrence of lichens

    Energy Technology Data Exchange (ETDEWEB)

    Kirschbaum, A; Klee, R

    1971-01-01

    Studies with lichen explants and with naturally occurring lichens, conducted in the Lower Main region in West Germany within the framework of an air hydgienic and meteorologic model study of that region, are described. Parmelia physodes explants from oak trees growing in nonpolluted areas were exposed in polluted areas, such as in an industrial area, an airport, a petroleum refinery, and near a large chemical plant. The degree of air pollution in the exposure site was evaluated by the degree of the lichen damage in seven grades. The large-scale average distribution of air pollution in the survey area was studied by surveying the natural occurrence of lichen species on 10 apple trees in area units of 6.25 sq km each. The lichen explant and lichen survey methods compared by the study of naturally occurring lichens were near the exposure site of lichen explants.

  20. Late graft explants in endovascular aneurysm repair.

    Science.gov (United States)

    Turney, Eric J; Steenberge, Sean P; Lyden, Sean P; Eagleton, Matthew J; Srivastava, Sunita D; Sarac, Timur P; Kelso, Rebecca L; Clair, Daniel G

    2014-04-01

    With more than a decade of use of endovascular aneurysm repair (EVAR), we expect to see a rise in the number of failing endografts. We review a single-center experience with EVAR explants to identify patterns of presentation and understand operative outcomes that may alter clinical management. A retrospective analysis of EVARs requiring late explants, >1 month after implant, was performed. Patient demographics, type of graft, duration of implant, reason for removal, operative technique, length of stay, complications, and in-hospital and late mortality were reviewed. During 1999 to 2012, 100 patients (91% men) required EVAR explant, of which 61 were placed at another institution. The average age was 75 years (range, 50-93 years). The median length of time since implantation was 41 months (range, 1-144 months). Explanted grafts included 25 AneuRx (Medtronic, Minneapolis, Minn), 25 Excluder (W. L. Gore & Associates, Flagstaff, Ariz), 17 Zenith (Cook Medical, Bloomington, Ind), 15 Talent (Medtronic), 10 Ancure (Guidant, Indianapolis, Ind), 4 Powerlink (Endologix, Irvine, Calif), 1 Endurant (Medtronic), 1 Quantum LP (Cordis, Miami Lakes, Fla), 1 Aorta Uni Iliac Rupture Graft (Cook Medical, Bloomington, Ind), and 1 homemade tube graft. Overall 30-day mortality was 17%, with an elective case mortality of 9.9%, nonelective case mortality of 37%, and 56% mortality for ruptures. Endoleak was the most common indication for explant, with one or more endoleaks present in 82% (type I, 40%; II, 30%; III, 22%; endotension, 6%; multiple, 16%). Other reasons for explant included infection (13%), acute thrombosis (4%), and claudication (1%). In the first 12 months, 23 patients required explants, with type I endoleak (48%) and infection (35%) the most frequent indication. Conversely, 22 patients required explants after 5 years, with type I (36%) and type III (32%) endoleak responsible for most indications. The rate of EVAR late explants has increased during the past decade at our

  1. Indução de calos embriogênicos em explantes de cupuaçuzeiro Induction of embryogenics calli in cupuassu explants

    Directory of Open Access Journals (Sweden)

    Maria das Graças Rodrigues Ferreira

    2004-08-01

    Full Text Available Objetivou-se a indução de calos embriogênicos em cupuaçuzeiro, em função do tipo de explante e meio de cultura. Foram testados como explantes, segmentos cotiledonares e eixos embrionários divididos em três partes: região da plúmula, radícula e hipocótilo. Os explantes foram cultivados em 2 diferentes meios de cultura: 1 MS suplementado com 2,4-D (1 mg L-1 e Cinetina (0,25 mg L-1; 2 MS acrescido de ANA (5 mg L-1 e Cinetina (0,25 mg L-1. Constatou-se que a região do hipocótilo foi a parte mais responsiva do eixo embrionário, formando calos com aspecto branco e friável. As auxinas testadas nos meios não estimularam o processo embriogênico em calos de cupuaçuzeiro.It was studied the induction of embryogenics calli in cupuassu, in function of kind of explant and culture medium. Cotyledons segments and embryonic axes were tested and divided in three parts: region of plumule, radicule and hypocotile. The explants were cultivated in two different culture medium: 1 MS supplemented with 2,4-D (1 mg L-1 and Kinetin (0,25 mg L-1; 2 MS supplemented with NAA (5 mg L-1 and Kinetin (0,25 mg L-1. The hypocotile region demonstrated to be more responsive segment of the embryonic axe, forming callus with white and friable aspect. No somatic embryogenesis was evidenced in callus of cupuassu with auxines testeds in the medium.

  2. Micropropagation of Araucaria excelsa R. Br. var. glauca Carrière from orthotropic stem explants.

    Science.gov (United States)

    Sarmast, Mostafa Khoshhal; Salehi, Hassan; Khosh-Khui, Morteza

    2012-07-01

    The objectives of the present work were in vitro propagation of Araucaria excelsa R. Br. var. glauca Carrière (Norfolk Island pine) with focus on the evaluation of the mean number of shoots per explant (MNS/E) and mean length of shoots per explants (MLS/E) produced by different parts of the orthotropic stem of A. excelsa R. Br. var. glauca in response to plant growth regulators. Norfolk Island pine axillary meristems responded very well to the 2-iso-pentenyl adenine (2iP) and thidiazuron (TDZ) levels. Explants taken from stem upper segments in the media containing 2iP had a higher MNS/E (3.47) and MLS/E (6.27 mm) in comparison to those taken from stem lower segments, which were 0.71 and 0.51 mm, respectively. Using 0.045 μM TDZ in the MS medium not only resulted in 4.60 MNS/E with 7.08 mm MLS/E but proliferated shoots showed a good performance as well. Investigating the best position of stem explant on mother plant as well as the best concentrations of growth regulators were performed which were useful for efficient micropropagation of this plant. Thirty three percent of explants were rooted in the MS medium containing 3 % sucrose, supplemented with 7.5 μM of both NAA and IBA for 2 weeks before transferring to a half strength MS medium without any growth regulator. Plantlets obtained were acclimatized and transferred to the greenhouse with less than 20 % mortality. This procedure considered the first successful report for regeneration and acclimatization of A. excelsa R. Br. var. glauca plantlet through main stem explants.

  3. NAA-Induced Direct Organogenesis from Female Immature Inflorescence Explants of Date Palm.

    Science.gov (United States)

    Khierallah, Hussam S M; Bader, Saleh M; Al-Khafaji, Makki A

    2017-01-01

    Micropropagation has great potential for the multiplication of female and male date palms of commercially grown cultivars by using inflorescences. This approach is simple, convenient, and much faster than the conventional method of using shoot-tip explants. We describe here a stepwise micropropagation procedure using inflorescence explants of Iraqi date palm cultivar Maktoom. Cultured explants were derived from 0.5-cm-long spike segments excised from 8 to 10-cm-long spathes. About 70% formed adventitious buds on Murashige and Skoog (MS) medium supplemented with 2 mg/L naphthalene acetic acid (NAA), 4 mg/L benzylaminopurine (BAP), and 40 g/L sucrose and maintained in the dark for 16 weeks before transferring to normal light conditions. The best multiplication rate was achieved with 3 mg/L 2ip and 2 mg/L; for shoot elongation, the best medium is MS containing 0.5 mg/L BAP, 0.5 mg/L 2ip, and 1 mg/L GA 3 . Well-developed shoots were cultured for rooting in half MS medium amended with 1 mg/L NAA and 45 g/L sucrose. Plantlets with well-developed roots were successfully hardened in the greenhouse. Inflorescence explants proved to be a promising alternative explant source for micropropagation of date palm cultivars.

  4. Citrus tissue culture employing vegetative explants.

    Science.gov (United States)

    Chaturvedi, H C; Singh, S K; Sharma, A K; Agnihotri, S

    2001-11-01

    Citrus being a number one fruit of the world due to its high nutritional value, huge production of fruits and fruit products, the citrus industry may be considered a major fruit industry. Though citrus orchard area in India is comparable to USA, the produce is far less, while its export is nil. Biotechnology has played an outstanding role in boosting the citrus industry, e.g., in Spain, which is now the biggest exporter of citrus fruit with the application of micrografting. Amongst the fruit trees, perhaps the maximum tissue culture research has been done in citrus during the past four decades, however, the results of practical value are meagre. The shortfalls in citrus tissue culture research and some advancements made in this direction along with bright prospects are highlighted, restricting the review to vegetative explants only. Whilst utilization of nucellar embryogenesis is limited to rootstocks, the other aspects, like, regeneration and proliferation of shoot meristems measuring 200 microm in length--a global breakthrough--of two commercially important scion species, Citrus aurantifolia and C. sinensis and an important rootstock, C. limonia, improvement of micrografting technique, cloning of the same two scion species as well as some Indian rootstock species, employing nodal stem segments of mature trees, of immense practical value have been elaborated. A rare phenomenon of shift in the morphogenetic pattern of differentiation from shoot bud differentiation to embryoid formation occurred during the long-term culture of stem callus of C. grandis. Stem callus-regenerated plants of C. aurantifolia, C. sinensis and C. grandis showed variation in their ploidy levels and a somaclonal variant of C. sinensis, which produced seedless fruits was isolated. Tailoring of rooting in microshoots to a tap root-like system by changing the inorganic salt composition of the rooting medium, resulting in 100% transplant success, and germplasm preservation through normal growth

  5. Effect of explant age, hormones on somatic embryogenesis and ...

    African Journals Online (AJOL)

    Effect of explant age, hormones on somatic embryogenesis and production of multiple shoot from cotyledonary leaf explants of Solanum trilobatum L. VNC Dhavala, RD Tejeswara, VR Yechuri, K Prabavathi ...

  6. Pengkajian Penerapan Teknik Budidaya Rhizophora mucronata dengan Stek Hipokotil (Study on Propagation Technique Application of Rhizophora mucronata Using Hypocotyl Cutting System

    Directory of Open Access Journals (Sweden)

    Neni Mulyani

    2007-01-01

    Full Text Available Research on the vegetative propagation of Rhizophora mucronata using hypocotyl cutting sistem was carried out at Faculty of Forestry, Bogor Agricultural University. Histological analysis of adventitous shoot was done at Biotechnology and Tree Breeding Laboratory, SEAMEO-BIOTROP. Three types of hypocotyl cutting (complete hypocotyl, top part of cut-hypocotyl and bottom part of cut-hypocotyl were induced using 6-BAP hormones at 0 and 500 ppm. Adventitous shoot of bottom part of cut-hypocotyl appeared 140 days after planting. Shoot from complete hypocotyl and top part of cut-hypocotyl grew earlier than adventitous shoot from bottom part of cut-hypocotyl. Hystologycal analysis of adventitous shoot showed the existance of primary meristem, stipule and leaf primordia. The growth of complete hipocotyl was higher than both of top part of cut-hypocotyl and bottom part of cut-hypocotyl. There was no effect of hormone induction on the shoot growth of them.

  7. In vitro clonal propagation of Achyranthes aspera L. and Achyranthes bidentata Blume using nodal explants.

    Science.gov (United States)

    Gnanaraj, Wesely Edward; Antonisamy, Johnson Marimuthu; R B, Mohanamathi; Subramanian, Kavitha Marappampalyam

    2012-01-01

    To develop the reproducible in vitro propagation protocols for the medicinally important plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blume using nodal segments as explants. Young shoots of A. aspera and A. bidentata were harvested and washed with running tap water and treated with 0.1% bavistin and rinsed twice with distilled water. Then the explants were surface sterilized with 0.1% (w/v) HgCl2 solutions for 1 min. After rinsing with sterile distilled water for 3-4 times, nodal segments were cut into smaller segments (1 cm) and used as the explants. The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), kinetin (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration. Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments inoculated on MS basal medium with 3% sucrose and augmented with BAP and Kin with varied frequency. MS medium augmented with 3.0 mg/L of BAP showed the highest percentage (93.60±0.71) of shootlets formation for A. aspera and (94.70±0.53) percentages for A. bidentata. Maximum number of shoots/explants (10.60±0.36) for A. aspera and (9.50±0.56) for A. bidentata was observed in MS medium fortified with 5.0 mg/L of BAP. For A. aspera, maximum mean length (5.50±0.34) of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A. bidentata (5.40±0.61) was observed in the very same concentration. The highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of IBA. Seventy percentages of plants were successfully established in polycups. Sixty eight percentages of plants were well established in the green house condition. Sixty five percentages of

  8. Effect of explant origin and different growth regulators on micropropagation of Pistacia atlantica ssp. mutica

    Directory of Open Access Journals (Sweden)

    Ali-Ashraf Mehrabi

    2015-06-01

    Full Text Available Propagation of wild pistachio as a multipurpose woody species is a hard and tedious task. In this research, an effective in vitro protocol was developed for rapid proliferation of wild pistachio (Pistacia atlantica ssp. mutica in MS medium supplemented with B5 vitamins and different growth regulators. Rooting of plantlets was tested by two treatments containing Rhizopon and IBA in ex vitro. With respect to the results, the nodal segments explants, produced the highest shoot frequency, leaf frequency and the tallest shoots. On the other hand, the tallest shoots were generated from shoot tip explant and medium containing of TDZ plus IAA. Both treatments (Rhizopon and IBA led to a remarkable increase in the number of roots, root length and rooting percentage compared to the control. These results may be applied for rapid proliferation to spread the pistachio trees and shrubs that are difficult and time consuming.

  9. Characterization and functional analysis of four HYH splicing variants in Arabidopsis hypocotyl elongation.

    Science.gov (United States)

    Li, Chen; Zheng, Lanlan; Zhang, Jingxuan; Lv, Yanxia; Liu, Jianping; Wang, Xuanbin; Palfalvi, Gergo; Wang, Guodong; Zhang, Yonghong

    2017-07-01

    Arabidopsis thaliana LONG HYPOCOTYL5 (HY5) is a positive regulator of the light signaling pathway. The hy5 mutant has an elongated hypocotyl in all light conditions, whereas the hy5 homolog (hyh) mutant has a very weak phenotype, but only in blue light. However, overexpression of HYH rescues the elongated hypocotyl phenotype in the hy5 null mutant. Here, we report the identification of four HYH splicing variants in Arabidopsis. Alternative splicing in the 5' region of the HYH gene occurred such that the proteins encoded by all four HYH variants retained their bZIP domain. In hypocotyl tissue, transcript levels of HYH.2, HYH.3, and HYH.4 were higher than those of HYH.1. Like HY5, all HYH variants were induced by light. Functional analysis of the four HYH variants, based on their abilities to complement the hy5 mutant, indicated that they have similar roles in hypocotyl development, and may function redundantly with HY5. Our results indicate that the bZIP domain in HYH is critical for the function of four variants in the compensation of hy5 mutant in hypocotyl development. Additionally, while HY5/HYH is found in plant species ranging from green algae to flowering plants, the potential alternative splicing events are distinct in different species, with certain HYH variants found with greater frequency in some species than others. Copyright © 2017. Published by Elsevier B.V.

  10. Autolysis and extension of isolated walls from growing cucumber hypocotyls

    Science.gov (United States)

    Cosgrove, D. J.; Durachko, D. M.

    1994-01-01

    Walls isolated from cucumber hypocotyls retain autolytic activities and the ability to extend when placed under the appropriate conditions. To test whether autolysis and extension are related, we treated the walls in various ways to enhance or inhibit long-term wall extension ('creep') and measured autolysis as release of various saccharides from the wall. Except for some non-specific inhibitors of enzymatic activity, we found no correlation between wall extension and wall autolysis. Most notably, autolysis and extension differed strongly in their pH dependence. We also found that exogenous cellulases and pectinases enhanced extension in native walls, but when applied to walls previously inactivated with heat or protease these enzymes caused breakage without sustained extension. In contrast, pretreatment of walls with pectinase or cellulase, followed by boiling in methanol to inactivate the enzymes, resulted in walls with much stronger expansin-mediated extension responses. Crude protein preparations from the digestive tracts of snails enhanced extension of both native and inactivated walls, and these preparations contained expansin-like proteins (assessed by Western blotting). Our results indicate that the extension of isolated cucumber walls does not depend directly on the activity of endogenous wall-bound autolytic enzymes. The results with exogenous enzymes suggest that the hydrolysis of matrix polysaccharides may not induce wall creep by itself, but may act synergistically with expansins to enhance wall extension.

  11. Stabilization of gene expression and cell morphology after explant recycling during fin explant culture in goldfish

    International Nuclear Information System (INIS)

    Chenais, Nathalie; Lareyre, Jean-Jacques; Le Bail, Pierre-Yves; Labbe, Catherine

    2015-01-01

    The development of fin primary cell cultures for in vitro cellular and physiological studies is hampered by slow cell outgrowth, low proliferation rate, poor viability, and sparse cell characterization. Here, we investigated whether the recycling of fresh explants after a first conventional culture could improve physiological stability and sustainability of the culture. The recycled explants were able to give a supplementary cell culture showing faster outgrowth, cleaner cell layers and higher net cell production. The cells exhibited a highly stabilized profile for marker gene expression including a low cytokeratin 49 (epithelial marker) and a high collagen 1a1 (mesenchymal marker) expression. Added to the cell spindle-shaped morphology, motility behavior, and actin organization, this suggests that the cells bore stable mesenchymal characteristics. This contrast with the time-evolving expression pattern observed in the control fresh explants during the first 2 weeks of culture: a sharp decrease in cytokeratin 49 expression was concomitant with a gradual increase in col1a1. We surmise that such loss of epithelial features for the benefit of mesenchymal ones was triggered by an epithelial to mesenchymal transition (EMT) process or by way of a progressive population replacement process. Overall, our findings provide a comprehensive characterization of this new primary culture model bearing mesenchymal features and whose stability over culture time makes those cells good candidates for cell reprogramming prior to nuclear transfer, in a context of fish genome preservation. - Highlights: • Recycled fin explants outgrow cells bearing stable mesenchymal traits. • Cell production and quality is enhanced in the recycled explant culture system. • Fresh fin primary culture is highly variable and loose epithelial traits over time

  12. Stabilization of gene expression and cell morphology after explant recycling during fin explant culture in goldfish

    Energy Technology Data Exchange (ETDEWEB)

    Chenais, Nathalie; Lareyre, Jean-Jacques; Le Bail, Pierre-Yves; Labbe, Catherine, E-mail: catherine.labbe@rennes.inra.fr

    2015-07-01

    The development of fin primary cell cultures for in vitro cellular and physiological studies is hampered by slow cell outgrowth, low proliferation rate, poor viability, and sparse cell characterization. Here, we investigated whether the recycling of fresh explants after a first conventional culture could improve physiological stability and sustainability of the culture. The recycled explants were able to give a supplementary cell culture showing faster outgrowth, cleaner cell layers and higher net cell production. The cells exhibited a highly stabilized profile for marker gene expression including a low cytokeratin 49 (epithelial marker) and a high collagen 1a1 (mesenchymal marker) expression. Added to the cell spindle-shaped morphology, motility behavior, and actin organization, this suggests that the cells bore stable mesenchymal characteristics. This contrast with the time-evolving expression pattern observed in the control fresh explants during the first 2 weeks of culture: a sharp decrease in cytokeratin 49 expression was concomitant with a gradual increase in col1a1. We surmise that such loss of epithelial features for the benefit of mesenchymal ones was triggered by an epithelial to mesenchymal transition (EMT) process or by way of a progressive population replacement process. Overall, our findings provide a comprehensive characterization of this new primary culture model bearing mesenchymal features and whose stability over culture time makes those cells good candidates for cell reprogramming prior to nuclear transfer, in a context of fish genome preservation. - Highlights: • Recycled fin explants outgrow cells bearing stable mesenchymal traits. • Cell production and quality is enhanced in the recycled explant culture system. • Fresh fin primary culture is highly variable and loose epithelial traits over time.

  13. Efficient regeneration of plants from shoot tip explants of ...

    African Journals Online (AJOL)

    Dendrobium densiflorum Lindl. is one of the horticulturally important orchids of Nepal due to its beautiful yellowish flower and medicinal properties. The present study was carried out for plant regeneration from shoot tip explants of D. densiflorum by tissue culture technique. The shoot tip explants of this species, obtained ...

  14. The effect of plant growth regulators, explants and cultivars on ...

    African Journals Online (AJOL)

    To achieve the best explants and media for spinach tissue culture, the effects of two different plant growth regulators, two explants and cultivars on adventitious shoot regeneration were tested. The Analysis of Variance (ANOVA) showed that the effects of plant growth regulators on spinach tissue culture were significant; ...

  15. Adventitious shoot regeneration from leaf explants of the valuable ...

    African Journals Online (AJOL)

    The objective of this study was to develop an efficient protocol for adventitious shoot regeneration for Plectranthus barbatus Andrews using leaf explants. The explants were cultured on MS (Murashige and Skoog, 1962) medium containing various concentration of kinetin (KN), 6-benzylaminopurine (BAP) and thidiazuron ...

  16. Studies on Callus Induction and Regeneration of Medicinal Plant Chicory (Cichorium intybus L. from Leaf and Petiole Explants

    Directory of Open Access Journals (Sweden)

    H. Hadizadeh

    2016-07-01

    Full Text Available Introduction: Chicory (Cichorium intybus L. belongs to Asteraceae family is commonly known as witloof chicory. The leaves and the roots of this medicinal plant are edible and commonly used as salad. Some varieties are also cultivated as coffee substitute after roasting the roots. All parts of the plant contain these volatile oils, with the majority of the toxic components concentrated in the plant's root. In folk medicine, the plant is used for the treatment of diarrhea, spleen enlargement, fever, and vomiting. Antihepatotoxic activity on damaged rat’s liver sections and anti-bacterial activity of this crop has been recently reported. In vitro regeneration from leaf explants with various hormonal combinations has been reported previously. Moreover, in vitro regeneration of Chicory from cotyledon explants using different combinations of plant growth regulators has been studied. Also, a protocol for the regeneration of plantlets from leaf and petiole explants of witloof chicory has been developed. The aim of the present investigation was optimization of callus induction and shoot regeneration from leaf and petiole tissues of Chicory (Esfahan genotype. Materials and Methods: In this investigation, Esfahan genotype was used for callus induction and direct shoot regeneration. Seeds were first washed with running tap water for 30 min then seeds were surface sterilized by dipping in 70% ethanol for 90 s and rinsed with sterile distilled water, followed by immersing in 5% sodium hypochlorite solution for 25 min and thereafter rinsed for 30 min with sterile distilled water. The basal medium used in this investigation was MS. For shoot regeneration, leaf and petiole explants (5 mm segments were excised from 4-week-old sterile seedlings and cultured on MS medium containing different combinations of NAA / BA and KIN / BA in two separate experiments. Experiments were performed factorial based on completely randomized design. Cultures were incubated at 25

  17. Simple explant culture of the embryonic chicken retina with long-term preservation of photoreceptors.

    Science.gov (United States)

    Thangaraj, Gopenath; Greif, Alexander; Layer, Paul G

    2011-10-01

    Structurally stable in vitro-model systems are indispensible to analyse neural development during embryogenesis, follow cellular differentiation and evaluate neurotoxicological or growth factor effects. Here we describe a three-dimensional, long-term in vitro-culture system of the embryonic chick retina which supports photoreceptor development. Retinal tissue was isolated from E6 chick eye, and cultured as explants by continuous orbital rotation to allow free floatation without any supporting materials. Young stage (E6) immature retinas were cultured for various time periods in order to follow the differentiation of cell types and plexiform layers by immunocytochemical methods. These explants could be cultured for at least 2-3 weeks with remarkable retention of retinal architecture. Interestingly, photoreceptors developed in the absence of pigment epithelium. Electron microscopic studies revealed formation of structures resembling photoreceptor outer segments, a feature not reported previously. Thus, the verification of photoreceptors, Müller cells, inner retinal cells and the inner plexiform layer described in our study establishes this explant culture as a valuable in vivo-like model system. Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.

  18. Studying secondary growth and bast fiber development: the hemp hypocotyl peeks behind the wall

    Directory of Open Access Journals (Sweden)

    Marc Behr

    2016-11-01

    Full Text Available Cannabis sativa L. is an annual herbaceous crop grown for the production of long extraxylary fibers, the bast fibers, rich in cellulose and used both in the textile and biocomposite sectors. Despite being herbaceous, hemp undergoes secondary growth and this is well exemplified by the hypocotyl. The hypocotyl was already shown to be a suitable model to study secondary growth in other herbaceous species, namely Arabidopsis thaliana and it shows an important practical advantage, i.e. elongation and radial thickening are temporally separated. This study focuses on the mechanisms marking the transition from primary to secondary growth in the hemp hypocotyl by analysing the suite of events accompanying vascular tissue and bast fiber development. Transcriptomics, imaging and quantification of phytohormones were carried out on four representative developmental stages (i.e. 6-9-15-20 days after sowing to provide a comprehensive overview of the events associated with primary and secondary growth in hemp. This multidisciplinary approach provides cell wall-related snapshots of the growing hemp hypocotyl and identifies marker genes associated with the young (expansins, β-galactosidases and transcription factors involved in light-related processes and the older hypocotyl (secondary cell wall biosynthetic genes and transcription factors.

  19. Stabilization of gene expression and cell morphology after explant recycling during fin explant culture in goldfish.

    Science.gov (United States)

    Chenais, Nathalie; Lareyre, Jean-Jacques; Le Bail, Pierre-Yves; Labbe, Catherine

    2015-07-01

    The development of fin primary cell cultures for in vitro cellular and physiological studies is hampered by slow cell outgrowth, low proliferation rate, poor viability, and sparse cell characterization. Here, we investigated whether the recycling of fresh explants after a first conventional culture could improve physiological stability and sustainability of the culture. The recycled explants were able to give a supplementary cell culture showing faster outgrowth, cleaner cell layers and higher net cell production. The cells exhibited a highly stabilized profile for marker gene expression including a low cytokeratin 49 (epithelial marker) and a high collagen 1a1 (mesenchymal marker) expression. Added to the cell spindle-shaped morphology, motility behavior, and actin organization, this suggests that the cells bore stable mesenchymal characteristics. This contrast with the time-evolving expression pattern observed in the control fresh explants during the first 2 weeks of culture: a sharp decrease in cytokeratin 49 expression was concomitant with a gradual increase in col1a1. We surmise that such loss of epithelial features for the benefit of mesenchymal ones was triggered by an epithelial to mesenchymal transition (EMT) process or by way of a progressive population replacement process. Overall, our findings provide a comprehensive characterization of this new primary culture model bearing mesenchymal features and whose stability over culture time makes those cells good candidates for cell reprogramming prior to nuclear transfer, in a context of fish genome preservation. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Agrobacterium rhizogenes-dependent production of transformed roots from foliar explants of pepper (Capsicum annuum): a new and efficient tool for functional analysis of genes.

    Science.gov (United States)

    Aarrouf, J; Castro-Quezada, P; Mallard, S; Caromel, B; Lizzi, Y; Lefebvre, V

    2012-02-01

    Pepper is known to be a recalcitrant species to genetic transformation via Agrobacterium tumefaciens. A. rhizogenes-mediated transformation offers an alternative and rapid possibility to study gene functions in roots. In our study, we developed a new and efficient system for A. rhizogenes transformation of the cultivated species Capsicum annuum. Hypocotyls and foliar organs (true leaves and cotyledons) of Yolo Wonder (YW) and Criollo de Morelos 334 (CM334) pepper cultivars were inoculated with the two constructs pBIN-gus and pHKN29-gfp of A. rhizogenes strain A4RS. Foliar explants of both pepper genotypes infected by A4RS-pBIN-gus or A4RS-pHKN29-gfp produced transformed roots. Optimal results were obtained using the combination of the foliar explants with A4RS-pHKN29-gfp. 20.5% of YW foliar explants and 14.6% of CM334 foliar explants inoculated with A4RS-pHKN29-gfp produced at least one root expressing uniform green fluorescent protein. We confirmed by polymerase chain reaction the presence of the rolB and gfp genes in the co-transformed roots ensuring that they integrated both the T-DNA from the Ri plasmid and the reporter gene. We also demonstrated that co-transformed roots of YW and CM334 displayed the same resistance response to Phytophthora capsici than the corresponding untransformed roots. Our novel procedure to produce C. annuum hairy roots will thus support the functional analysis of potential resistance genes involved in pepper P. capsici interaction.

  1. Exogenous transforming growth factor-β1 enhances smooth muscle differentiation in embryonic mouse jejunal explants.

    Science.gov (United States)

    Coletta, Riccardo; Roberts, Neil A; Randles, Michael J; Morabito, Antonino; Woolf, Adrian S

    2017-01-13

    An ex vivo experimental strategy that replicates in vivo intestinal development would in theory provide an accessible setting with which to study normal and dysmorphic gut biology. The current authors recently described a system in which mouse embryonic jejunal segments were explanted onto semipermeable platforms and fed with chemically defined serum-free media. Over 3 days in organ culture, explants formed villi and they began to undergo spontaneous peristalsis. As defined in the current study, the wall of the explanted gut failed to form a robust longitudinal smooth muscle (SM) layer as it would do in vivo over the same time period. Given the role of transforming growth factor β1 (TGFβ1) in SM differentiation in other organs, it was hypothesized that exogenous TGFβ1 would enhance SM differentiation in these explants. In vivo, TGFβ receptors I and II were both detected in embryonic longitudinal jejunal SM cells and, in organ culture, exogenous TGFβ1 induced robust differentiation of longitudinal SM. Microarray profiling showed that TGFβ1 increased SM specific transcripts in a dose dependent manner. TGFβ1 proteins were detected in amniotic fluid at a time when the intestine was physiologically herniated. By analogy with the requirement for exogenous TGFβ1 for SM differentiation in organ culture, the TGFβ1 protein that was demonstrated to be present in the amniotic fluid may enhance intestinal development when it is physiologically herniated in early gestation. Future studies of embryonic intestinal cultures should include TGFβ1 in the defined media to produce a more faithful model of in vivo muscle differentiation. Copyright © 2017 The Authors Journal of Tissue Engineering and Regenerative Medicine Published by John Wiley & Sons, Ltd. Copyright © 2017 The Authors Journal of Tissue Engineering and Regenerative Medicine Published by John Wiley & Sons, Ltd.

  2. Response of pine hypocotyl sections to growth regulators and related substances

    Directory of Open Access Journals (Sweden)

    J. Zakrzewski

    2015-01-01

    Full Text Available Growth response of Pinus silvestris hypocotyl sections to some synthetic growth regulators and related substances was studied. Elongation of hypocotyl sections was stimulated by naphtaleneacetic acid, indole-3-acetic acid, in-dole-3-propionic acid, indole-3-butyric acid, 2,4-dichlorophenoxyacetic acid, indoleaoetic amide, indoleacetic nitrile and coumarin. Indole-3-acetic acid and naphtaleneacetic acid extended period of growth up to 16 and 24 hours, respectively. Growth was inhibited by kinetin, trans-cinnamic acid and 2,3,5-tri-iodobenzoic acid. No effect of gibberellic acid, tryptophan and biotin was observed.

  3. Making post-mortem implantable cardioverter defibrillator explantation safe

    DEFF Research Database (Denmark)

    Räder, Sune B E W; Zeijlemaker, Volkert; Pehrson, Steen

    2009-01-01

    that the resting voltage over the operating person would not exceed 50 V. CONCLUSION: The use of intact medical gloves made of latex, neoprene, or plastic eliminates the potential electrical risk during explantation of an ICD. Two gloves on each hand offer sufficient protection. We will recommend the use......AIMS: The aim of this study is to investigate whether protection with rubber or plastic gloves during post-mortem explantation of an implantable cardioverter defibrillator (ICD) offers enough protection for the explanting operator during a worst-case scenario (i.e. ICD shock). METHODS AND RESULTS...

  4. Propagation of goldenrod (Solidago canadensis L. from leaf and nodal explants

    Directory of Open Access Journals (Sweden)

    Jun Li

    2012-02-01

    Full Text Available Goldenrod (Solidago canadensis L. is an invasive plant species in many countries except North America but a cut-flower species worldwide. There is a need to generate and propagate goldenrod clones efficiently for research and commercial purposes. A callus induction and plantlet regeneration system was developed by studying the influence of explant type and different concentrations of plant growth regulators. The highest callus production from leaf segments was obtained on Murashige and Skoog’s medium (MS medium supplemented with 1.0 mg/L naphthalene acetic acid (NAA and 1.0 mg/L 6-benzylaminopurine (BA. Adventitious shoots could be regenerated directly from leaf explants without an intermediate callus phase with the highest shoot induction percentage of 87.2%. The largest number of adventitious shoots per leaf explant (3.2 was obtained on MS medium supplemented with 0.4 mg/L NAA and 2.0 mg/L BA. MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L BA was the best medium for axillary shoot regeneration from nodal segments. The highest root number and longest roots occurred on half-strength MS without the addition of any growth regulator. Rooted plantlets were then transferred to a soil-based growth medium, placed in a greenhouse, and acclimatized with 100% success. All surviving plants grew normally without showing any morphological varia­tion when compared to those grow from seed. This regeneration protocol may be used to produce certain biotypes of goldenrod suitable for genetic transformation rapid propagation of goldenrod for commercial purposes or for screening fungi and toxins as potential biocontrol agents against this weed.

  5. Behavioural properties of chick somitic mesoderm and lateral plate when explanted in vitro.

    Science.gov (United States)

    Bellairs, R; Sanders, E J; Portch, P A

    1980-04-01

    Tissue culture, time-lapse cinematographic and electron microscopic techniques have been used to study the properties of chick mesoderm at several stages of differentiation. Lateral plate, unsegmented mesoderm (segmental plate), and newly formed somites were dissected from stage-12 embryos, whilst dermo-myotomes and sclerotomes were dissected from stage-18 embryos. Each type of mesoderm was found to exhibit a characteristic pattern of behaviour. The explants from the unsegmented mesoderm from the newly formed somites and from the older embryos could be placed in a developmental sequence; with increasing differentiation they settled and spread on the substrate more readily, whether explanted as pieces of tissue or as individual cells, and it was concluded that this implied an increased adhesion to the substrate. Similarly, with increasing differentiation, the cells segmented at a faster rate. No significant differences could be discerned in the internal structure of the different types of cells, although differences in the general shape were apparent. The lateral plate mesoderm cells, which bear some resemblances to the unsegmented mesoderm cells in the embryo, also show some morphological resemblances to them in vitro. However, the lateral plate cells had a much greater success in attaching to glass or platic substrates. They were also found to have the highest speed of locomotion of all the tissues studied, whereas the unsegmented had the lowest. It is concluded therefore, that although cells may look similar to one another morphologically, their behaviour may differ greatly, probably because they are already partially determined.

  6. Dedifferentiation of leaf explants and antileukemia activity of an ...

    African Journals Online (AJOL)

    user

    2011-04-04

    Apr 4, 2011 ... known as drumstick tree or horseradish tree (Little and. Wadsworth, 1964; Morton ... Leaves explants (Figure 1a) obtained from 21 day-old seedlings were sterilized by sodium ... Sweden), according to Harbeck et al. (1982).

  7. In vitro regeneration from internodal explants of bitter melon ...

    African Journals Online (AJOL)

    Thiru

    2012-04-24

    Apr 24, 2012 ... shoots per internodal explant after 80 days of culture. Key words: ... grown in the tropical regions of Asia, Amazon, east Africa and the ... Tamilnadu, India. .... expressed as the mean ± standard error (SE) of three experiments.

  8. Simple, effective and economical explant-surface sterilization ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-19

    Oct 19, 2009 ... recommend this technique due to its simplicity and economy. Key words: Explant ... mercuric chloride, hydrogen peroxide, silver nitrate and bromine water (Rai ... actually within the structure that is being surface steri- lized.

  9. Optimization of callus induction of Zataria multiflora under the effect of different plant growth regulators and explant source

    Directory of Open Access Journals (Sweden)

    N. Mosavat*

    2017-11-01

    Full Text Available Background and objectives: The Lamiaceae family is rich in favorable secondary metabolites which have different medicinal properties and also use in food, cosmetic and sanitary industry. Zataria multiflora Boiss. is an aromatic and bushy plant containing specific pharmaceutical components which is only distributed in certain regions of Iran. Tissue culture technologies could be suitable for in vitro production of Zataria. Methods: In this study, callus production and callus related traits of Zataria was evaluated at in vitro condition. Callus induction was performed on Murashige and Skoog (MS medium containing different levels of plant growth regulators including different cytokinins (Kinetin, benzyl amino purine and auxins (2,4 dichlorophenoxyacetic acid and naphtalen acetic acid and two different explant (hypocotyl and leaf. Results: The friable calli with yellow-green color only appeared from leaf explants on three different treatments including: 1: 2.5 (mg/L 2,4-D; 2: 2 (mg/L  2,4-D; 3: [2 (mg/L 2,4-D+ 1 (mg/L Kin].  The best callus induction (75% was obtained at 2,4-D (2 mg/L + Kin (1 mg/L after 2 month of incubation under the photoperiod of 16/8 (light/dark. The highest callus growth rate (CGR (0.072 mm/day and callus fresh weight (0.135 g were denoted to the treatment of 2 mg/L (2,4-D.  Conclusion: The benefits of the protocol described here include the possibility of its use throughout the callus culture for commercial production of suitable secondary metabolites of Zataria in rapid time and huge scale.

  10. An ovine tracheal explant culture model for allergic airway inflammation

    Directory of Open Access Journals (Sweden)

    Abeynaike Latasha

    2010-08-01

    Full Text Available Abstract Background The airway epithelium is thought to play an important role in the pathogenesis of asthmatic disease. However, much of our understanding of airway epithelial cell function in asthma has been derived from in vitro studies that may not accurately reflect the interactive cellular and molecular pathways active between different tissue constituents in vivo. Methods Using a sheep model of allergic asthma, tracheal explants from normal sheep and allergic sheep exposed to house dust mite (HDM allergen were established to investigate airway mucosal responses ex vivo. Explants were cultured for up to 48 h and tissues were stained to identify apoptotic cells, goblet cells, mast cells and eosinophils. The release of cytokines (IL-1α, IL-6 and TNF-α by cultured tracheal explants, was assessed by ELISA. Results The general morphology and epithelial structure of the tracheal explants was well maintained in culture although evidence of advanced apoptosis within the mucosal layer was noted after culture for 48 h. The number of alcian blue/PAS positive mucus-secreting cells within the epithelial layer was reduced in all cultured explants compared with pre-cultured (0 h explants, but the loss of staining was most evident in allergic tissues. Mast cell and eosinophil numbers were elevated in the allergic tracheal tissues compared to naïve controls, and in the allergic tissues there was a significant decline in mast cells after 24 h culture in the presence or absence of HDM allergen. IL-6 was released by allergic tracheal explants in culture but was undetected in cultured control explants. Conclusions Sheep tracheal explants maintain characteristics of the airway mucosa that may not be replicated when studying isolated cell populations in vitro. There were key differences identified in explants from allergic compared to control airways and in their responses in culture for 24 h. Importantly, this study establishes the potential for the

  11. Annuals sprouting adventitiously from the hypocotyl: their compensatory growth and implications for weed management

    Czech Academy of Sciences Publication Activity Database

    Latzel, Vít; Dospělová, L.; Klimešová, Jitka

    2009-01-01

    Roč. 64, č. 5 (2009), s. 923-929 ISSN 0006-3088 R&D Projects: GA ČR GD206/08/H044 Institutional research plan: CEZ:AV0Z60050516 Keywords : hypocotyle * regeneration * annuals Subject RIV: EF - Botanics Impact factor: 0.617, year: 2009

  12. Neighbor Detection Induces Organ-Specific Transcriptomes, Revealing Patterns Underlying Hypocotyl-Specific Growth.

    Science.gov (United States)

    Kohnen, Markus V; Schmid-Siegert, Emanuel; Trevisan, Martine; Petrolati, Laure Allenbach; Sénéchal, Fabien; Müller-Moulé, Patricia; Maloof, Julin; Xenarios, Ioannis; Fankhauser, Christian

    2016-12-01

    In response to neighbor proximity, plants increase the growth of specific organs (e.g., hypocotyls) to enhance access to sunlight. Shade enhances the activity of Phytochrome Interacting Factors (PIFs) by releasing these bHLH transcription factors from phytochrome B-mediated inhibition. PIFs promote elongation by inducing auxin production in cotyledons. In order to elucidate spatiotemporal aspects of the neighbor proximity response, we separately analyzed gene expression patterns in the major light-sensing organ (cotyledons) and in rapidly elongating hypocotyls of Arabidopsis thaliana PIFs initiate transcriptional reprogramming in both organs within 15 min, comprising regulated expression of several early auxin response genes. This suggests that hypocotyl growth is elicited by both local and distal auxin signals. We show that cotyledon-derived auxin is both necessary and sufficient to initiate hypocotyl growth, but we also provide evidence for the functional importance of the local PIF-induced response. With time, the transcriptional response diverges increasingly between organs. We identify genes whose differential expression may underlie organ-specific elongation. Finally, we uncover a growth promotion gene expression signature shared between different developmentally regulated growth processes and responses to the environment in different organs. © 2016 American Society of Plant Biologists. All rights reserved.

  13. Detection of endogenous gibberellins and their relationship to hypocotyl elongation in soybean seedlings

    International Nuclear Information System (INIS)

    Bensen, R.J.; Beall, F.D.; Mullet, J.E.; Morgan, P.W.

    1990-01-01

    Four gibberellins, GA 53 , GA 19 , GA 20 , and GA 1 , were detected by bioassay, chromatography in two HPLC systems, and combined gas chromatography-mass spectroscopy-selected ion monitoring (GC-MS-SIM) in etiolated soybean (Glycine max [L.] Merr.) hypocotyls. GC-MS-SIM employed [ 2 H 2 ]-labeled standards for each endogenous gibberellin detected, and quantities estimated from bioassays and GC-MS-SIM were similar. This result plus the tentative detection of GA 44 and GA 8 (standards not available) indicates that the early-C-13-hydroxylation pathway for gibberellin biosynthesis predominates in soybean hypocotyls. Other gibberellins were not detected. Growth rates decreased after transfer to low water potential (ψ w ) vermiculite and were completely arrested 24 hours after transfer. The GA 1 content in the elongating region of hypocotyls had declined to 38% of the 0 time value at 24 hours after transfer to low ψ w vermiculite, a level which was only 13% of the GA 1 content in control seedlings at the same time (24 hours posttransfer). Seedlings were growth responsive to exogenous GA 3 , and this GA 3 -promoted growth was inhibited by exogenous ABA. The data are consistent with the hypothesis that changes in GA 1 and ABA levels play a role in adjusting hypocotyl elongation rates. However, the changes observed are not of sufficient magnitude nor do they occur rapidly enough to suggest they are the primary regulators of elongation rate responses to rapidly changing plant water status

  14. Origin, timing, and gene expression profile of adventitious rooting in Arabidopsis hypocotyls and stems.

    Science.gov (United States)

    Welander, Margareta; Geier, Thomas; Smolka, Anders; Ahlman, Annelie; Fan, Jing; Zhu, Li-Hua

    2014-02-01

    Adventitious root (AR) formation is indispensable for vegetative propagation, but difficult to achieve in many crops. Understanding its molecular mechanisms is thus important for such species. Here we aimed at developing a rooting protocol for direct AR formation in stems, locating cellular AR origins in stems and exploring molecular differences underlying adventitious rooting in hypocotyls and stems. In-vitro-grown hypocotyls or stems of wild-type and transgenic ecotype Columbia (Col-0) of Arabidopsis thaliana were rooted on rooting media. Anatomy of AR formation, qRT-PCR of some rooting-related genes and in situ GUS expression were carried out during rooting from hypocotyls and stems. We developed a rooting protocol for AR formation in stems and traced back root origins in stems by anatomical and in situ expression studies. Unlike rooting in hypocotyls, rooting in stems was slower, and AR origins were mainly from lateral parenchyma of vascular bundles and neighboring starch sheath cells as well as, to a lesser extent, from phloem cap and xylem parenchyma. Transcript levels of GH3-3, LBD16, LBD29, and LRP1 in hypocotyls and stems were similar, but transcript accumulation was delayed in stems. In situ expression signals of DR5::GUS, LBD16::GUS, LBD29::GUS, and rolB::GUS reporters in stems mainly occurred at the root initiation sites, suggesting their involvement in AR formation. We have developed an efficient rooting protocol using half-strength Lepoivre medium for studying AR formation in stems, traced back the cellular AR origins in stems, and correlated expression of rooting-related genes with root initiation sites.

  15. A novel method for coral explant culture and micropropagation.

    Science.gov (United States)

    Vizel, Maya; Loya, Yossi; Downs, Craig A; Kramarsky-Winter, Esti

    2011-06-01

    We describe here a method for the micropropagation of coral that creates progeny from tissue explants derived from a single polyp or colonial corals. Coral tissue explants of various sizes (0.5-2.5 mm in diameter) were manually microdissected from the solitary coral Fungia granulosa. Explants could be maintained in an undeveloped state or induced to develop into polyps by manipulating environmental parameters such as light and temperature regimes, as well as substrate type. Fully developed polyps were able to be maintained for a long-term in a closed sea water system. Further, we demonstrate that mature explants are also amenable to this technique with the micropropagation of second-generation explants and their development into mature polyps. We thereby experimentally have established coral clonal lines that maintain their ability to differentiate without the need for chemical induction or genetic manipulation. The versatility of this method is also demonstrated through its application to two other coral species, the colonial corals Oculina patigonica and Favia favus.

  16. Utilization of Aseptic Seedling Explants for In vitro Propagation of Indian Red Wood

    Directory of Open Access Journals (Sweden)

    Kishore Kumar CHIRUVELLA

    2013-12-01

    Full Text Available Micropropagation has been advocated as one of the most viable biotechnological tool for ex situ conservation of rare, endangered endemic medicinal plants germplasm. Rapid clonal micropropagation protocol for large-scale multiplication of an endemic medicinal plant Soymida febrifuga (Meliaceae was established from 15-day aseptic seedling cotyledonary node and shoot tip explants. High frequency of sprouting and shoot differentiation was observed from cotyledonary node explants compared to shoot tip, on Murashige and Skoog (MS medium fortified with BA, KN, 2-iP and CM. Of the cytokinins used, BA (3.0 mgl-1 supported highest average number and maximum multiple shoot differentiation (16.6. In vitro proliferated shoots were multiplied rapidly by culturing nodal segments as microcuttings, further subcultured on the same media for elongation. Elongated shoots upon transfer to MS medium fortified with IBA showed rooting within two weeks of culture. Rooted plantlets were successfully hardened and 75% of rooted shoots successfully survived on establishment to the soil. Plants looked healthy with no visually detectable phenotypic variations. This protocol provides a successful and rapid technique that can be used for ex situ conservation minimizing the pressure on wild populations and contributes to the conservation of this endemic medicinally potent flora.

  17. Restricted cell elongation in Arabidopsis hypocotyls is associated with a reduced average pectin esterification level.

    Science.gov (United States)

    Derbyshire, Paul; McCann, Maureen C; Roberts, Keith

    2007-06-17

    Cell elongation is mainly limited by the extensibility of the cell wall. Dicotyledonous primary (growing) cell walls contain cellulose, xyloglucan, pectin and proteins, but little is known about how each polymer class contributes to the cell wall mechanical properties that control extensibility. We present evidence that the degree of pectin methyl-esterification (DE%) limits cell growth, and that a minimum level of about 60% DE is required for normal cell elongation in Arabidopsis hypocotyls. When the average DE% falls below this level, as in two gibberellic acid (GA) mutants ga1-3 and gai, and plants expressing pectin methyl-esterase (PME1) from Aspergillus aculeatus, then hypocotyl elongation is reduced. Low average levels of pectin DE% are associated with reduced cell elongation, implicating PMEs, the enzymes that regulate DE%, in the cell elongation process and in responses to GA. At high average DE% other components of the cell wall limit GA-induced growth.

  18. Effects of ionizing radiation on the hypocotyl-root axis of three species of gossypium

    International Nuclear Information System (INIS)

    Reed, J.P.

    1977-01-01

    The hypocotyl-root axis of cotton seedlings grown from irradiated and non-irradiated seeds was investigated using light microscopy and histological techniques. Special emphasis was placed on the pattern of vascular transition. Two patterns of vascular transition in non-irradiated seedlings were found. In Gossypium hirsutum and G. barbadense there are prominent metaxylem bands between the vascular bundles in the hypocotyl. In G. gossypioides there are no bands. The presence or absence of the bands was easily detected using polarized light. The most outstanding effects of radiation were inhibition of lateral root development and alteration of the pattern of vascular transition in seedlings grown from irradiated seeds. The findings suggest that the root apical meristem determines the vascular pattern

  19. Biosynthesis of phenolic compounds in hypocotyl callus cultures of fenugreek (Trigonella foenum graecum L. )

    Energy Technology Data Exchange (ETDEWEB)

    Dhandapani, M; Antony, A; Subba Rao, P V [Indian Inst. of Science, Bangalore. Dept. of Biochemistry

    1977-03-01

    Hypocotyl callus cultures of fenugreek were studied to determine their potential for synthesizing phenolics, particularly those which are intermediates in lignin and flavonoid biosynthesis. The cultures were found to be capable of synthesizing an array of phenolic compounds characteristic of higher plants. Both phenylalanine-U-/sup 14/C and cinnamic acid-U-/sup 14/C were found to be efficient precursors of these phenolics.

  20. Disruption of the actin cytoskeleton results in the promotion of gravitropism in inflorescence stems and hypocotyls of Arabidopsis

    Science.gov (United States)

    Yamamoto, Kazuyoshi; Kiss, John Z.

    2002-01-01

    The actin cytoskeleton is hypothesized to play a major role in gravity perception and transduction mechanisms in roots of plants. To determine whether actin microfilaments (MFs) are involved in these processes in stem-like organs, we studied gravitropism in Arabidopsis inflorescence stems and hypocotyls. Localization studies using Alexa Fluor-phalloidin in conjugation with confocal microscopy demonstrated a longitudinally and transversely oriented actin MF network in endodermal cells of stems and hypocotyls. Latrunculin B (Lat-B) treatment of hypocotyls caused depolymerization of actin MFs in endodermal cells and a significant reduction of hypocotyl growth rates. Actin MFs in Lat-B-treated inflorescence stems also were disrupted, but growth rates were not affected. Despite disruption of the actin cytoskeleton in these two organs, Lat-B-treated stems and hypocotyls exhibited a promotion of gravitropic curvature in response to reorientation. In contrast, Lat-B reduced gravitropic curvature in roots but also reduced the growth rate. Thus, in contrast to prevailing hypotheses, our results suggest that actin MFs are not a necessary component of gravitropism in inflorescence stems and hypocotyls. Furthermore, this is the first study to demonstrate a prominent actin MF network in endodermal cells in the putative gravity-perceiving cells in stems.

  1. The role of wall calcium in the extension of cell walls of soybean hypocotyls

    Science.gov (United States)

    Virk, S. S.; Cleland, R. E.

    1990-01-01

    Calcium crosslinks are load-bearing bonds in soybean (Glycine max (L.) Merr.) hypocotyl cell walls, but they are not the same load-bearing bonds that are broken during acid-mediated cell elongation. This conclusion is reached by studying the relationship between wall calcium, pH and the facilitated creep of frozen-thawed soybean hypocotyl sections. Supporting data include the following observations: 1) 2-[(2-bis-[carboxymethyl]amino-5-methylphenoxy)methyl]-6-methoxy-8-bis[car boxymethyl]aminoquinoline (Quin 2) and ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) caused only limited facilitated creep as compared with acid, despite removal of comparable or larger amounts of wall calcium; 2) the pH-response curves for calcium removal and acid-facilitated creep were different; 3) reversible acid-extension occurred even after removal of almost all wall calcium with Quin 2; and 4) growth of abraded sections did not involve a proportional loss of wall calcium. Removal of wall calcium, however, increased the capacity of the walls to undergo acid-facilitated creep. These data indicate that breakage of calcium crosslinks is not a major mechanism of cell-wall loosening in soybean hypocotyl tissues.

  2. Immunocytochemical characterization of explant cultures of human prostatic stromal cells

    NARCIS (Netherlands)

    A. Kooistra (Anko); A.M.J. Elissen (Arianne ); J.J. Konig (Josee); M. Vermey; Th.H. van der Kwast (Theo); J.C. Romijn (Johannes); F.H. Schröder (Fritz)

    1995-01-01

    textabstractThe study of stromal-epithelial interactions greatly depends on the ability to culture both cell types separately, in order to permit analysis of their interactions under defined conditions in reconstitution experiments. Here we report the establishment of explant cultures of human

  3. High frequency plant regeneration from shoot tip explants of ...

    African Journals Online (AJOL)

    USER

    2010-08-02

    Aug 2, 2010 ... 16/8-h (light/dark) photoperiod provided by cool white fluorescent light. Multiple .... formation from shoot tip explant of C. colocynthis on MS-medium. S. No. .... micropropagation of Musa sapientum L. (Cavendish Dwarf). Afr. J.

  4. Effect of season, explants, growth regulators and sugar level on ...

    African Journals Online (AJOL)

    Administrator

    2011-06-06

    Jun 6, 2011 ... swabbed with an alcohol (50% v/v) soaked muslin cloth followed by washing in ... water. The explants were trimmed to a final size of 0.5 to 2 cm and were placed .... microbial contaminants so that a specific treatment may be formulated for ... of 2,4-D causing significantly delayed response, for all the three ...

  5. callus induction and proliferation from cotyledon explants in ...

    African Journals Online (AJOL)

    ACSS

    2013-07-19

    Jul 19, 2013 ... between the tested cytokinins and the 2,4-D in callus induction and growth index. Similar results have also been obtained in soybean (Glycine max L.) (Sairam et al., 2003). In contrast to this finding, calli were reportedly induced from cotyledon explants on MS basal medium containing 2,4-D in combination ...

  6. Explant age, auxin concentrations and media type affect callus ...

    African Journals Online (AJOL)

    The effects of explant age of oil palm (Elaeis huineensis) embryo axes, 15 and 18 weeks after anthesis (WAA), media type (Eeuwens and Murashige and Skoog) supplemented with various concentrations of 2,4-D on callus production employing standard in vitro techniques were investigated. The results of the study showed ...

  7. The effect of plant growth regulators, explants and cultivars on ...

    African Journals Online (AJOL)

    ONOS

    2010-07-05

    Jul 5, 2010 ... The effect of plant growth regulators, explants and cultivars on spinach (Spinacia oleracea L.) tissue culture. Taha Roodbar Shojaei1*, Vahid Salari2, Darioush Ramazan3, Mahdi Ehyaei1, Javad. Gharechahi4 and Roya Motallebi Chaleshtori5. 1Department of Agronomy and Plant Breeding, College of ...

  8. 143 GROWTH RESPONSE OF EXPLANTS OF Irvingia gabonensis ...

    African Journals Online (AJOL)

    1&5 Department of Plant Science and Biotechnology, University of Port Harcourt. 2&3Department of Botany, University of Calabar. 4Department of Biological Sciences, Cross River State University of Technology. ABSTRACT. Growth response of explants of Irvingia gabonensis to in vitro treatment was investigated using full ...

  9. Culturing bovine nucleus pulposus explants by balancing medium osmolarity

    NARCIS (Netherlands)

    Dijk, van B.G.M.; Potier, E.; Ito, K.

    2011-01-01

    Regenerative therapies are promising treatments for early intervertebral disc degeneration. To test their efficacy, an in vitro tissue-level model would be valuable. Nucleus pulposus (NP) explant culture may constitute such a model, as the earliest signs of degeneration are in the NP. However, in NP

  10. Adventitious shoot regeneration from leaf explants of the valuable ...

    African Journals Online (AJOL)

    Jane

    2011-08-10

    Aug 10, 2011 ... 2Department of Plant Biology and Plant Biotechnology, St. Joseph's College, Tiruchirappalli – 620 002, South India. Accepted 28 March, 2011 .... Effect of cytokinins (KN and BAP) alone or in combination with NAA on direct shoot bud regeneration from leaf explants of P. barbatus. Plant growth regulator ...

  11. [Reasons for exchange and explantation of intraocular lenses].

    Science.gov (United States)

    Neuhann, I; Fleischer, F; Neuhann, T

    2012-08-01

    This study was performed to analyse the reasons for explantation/exchange of intraocular lenses (IOL), which had originally been implanted for the correction of aphakia during cataract extraction. All cases with IOL explantation, which had been performed at one institution between 1/2008 and 12/2009 were analysed retrospectively. A total of 105 eyes of 100 patients were analysed. The median time interval between implantation and explantation of the IOL was 5.9 years (min. 0, max. 29.6). The most frequent cause for the intervention was subluxation/dislocation of the implant in 55.2% of cases. This group comprised 21% of cases with subluxation within the capsular bag in pseudoexfoliation syndrome. Other reasons were optical problems/incorrect IOL power (21%), calcification of hydrophilic acrylic IOL (7.6%), corneal decompensation associated with an anterior chamber lens (4.8%), and single cases with varying problems. The reasons for IOL exchange presented in this study are comparable to those of other series in the literature. Explantations due to optical problems may gain weight in the future due to a rise in refractive procedures and demands. © Georg Thieme Verlag KG Stuttgart · New York.

  12. Somatic embryogenesis and plant regeneration from leaf explants of ...

    African Journals Online (AJOL)

    An attempt was made to study the somatic embryogenesis and plant regeneration from the in vitro leaf explants of Rumex vesicarius L. a renowned medicinal plant, which belongs to polygonaceae family. Effective in vitro regeneration of R. vesicarius was achieved via young leaf derived somatic embryo cultures.

  13. Plant regeneration via somatic embryogenesis from root explants of ...

    African Journals Online (AJOL)

    A system for induction of callus and plant regeneration via somatic embryogenesis from root explants of Hevea brasiliensis Muell. Arg. clone Reyan 87-6-62 was evaluated. The influence of plant growth regulators (PGRs) including 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (6-BA) and kinetin (KT) on ...

  14. Absorção de macronutrientes por explantes de bananeira in vitro Macronutrient absorption by banana explants in vitro

    Directory of Open Access Journals (Sweden)

    Josefa Diva Nogueira Diniz

    1999-07-01

    Full Text Available Com o objetivo de estudar a absorção de macronutrientes (N, P, K, Ca, Mg e S em explantes de bananeira cv. Prata Anã, foram utilizados explantes de plantas estabelecidas in vitro, inoculados em meio básico de Murashige & Skoog (1962 contendo sacarose (30 g/L, e BAP (3,5 mg/L com sete tratamentos, representados pelos períodos de 0, 10, 20, 30, 40, 50 e 60 dias de cultivo e três repetições. As quantidades de macronutrientes totais absorvidas pelos explantes seguiram a ordem: K > N > Ca > ou = P > Mg @ S. O P foi o nutriente absorvido mais rapidamente pelos explantes, com 75% extraído do meio de cultivo nos primeiros 30 dias, cessando sua absorção aos 50 dias, restando ainda 9% no meio de cultivo. A absorção do S cessou também aos 50 dias, quando 66% deste nutriente ainda permanecia no meio de cultivo. Este resultado sugere haver uma relação, quanto à absorção, entre esses dois nutrientes. As maiores taxas de absorção de todos os nutrientes foram verificadas nos primeiros 20 dias. O rizoma, o pseudocaule e as folhas, se diferenciaram quanto à concentração e extração ou acúmulo de nutrientes.The absorption of the nutrients (N, P, K, Ca, Mg and S by banana (Musa sp. cv. Prata Anã explants on the basic medium of Murashige & Skoog (1962 supplemented with sucrose (30 g/L and BAP (3.5 mg/L were evaluated at 0, 10, 20, 30, 40, 50 and 60 days after inoculation. The seven treatments were arranged on a completely randomized design with three replicates. The sequence of nutrient absorption by the explants was K > N > Ca > or = P > Mg @ S. The P was the nutrient with the fastest absorption rate and at the 30th day the explants had already absorbed 75% of the P from the medium. The P absorption stopped by the 50th day. The S absorption stopped at the 50th day with 66% of it remaining in the medium. The results suggested a close relationship between these two nutrients. The highest rates of nutrient absorption were observed during the

  15. Effect of storage media and time on fin explants culture in the ...

    African Journals Online (AJOL)

    The effect of storage media and time was investigated on fin explants culture in the goldfish (Carassius auratus). Fin explants under sterile conditions were able to produce cells at different storage media and time. On the outgrowth of cells, fin explants stored for seven days before culturing showed significantly higher growth ...

  16. Sealing and explant types on the mangaba micropropagation Tipo de vedação e explantes na micropropagação de mangabeira

    Directory of Open Access Journals (Sweden)

    Aline de Jesus Sá

    2012-08-01

    Full Text Available In micropropagation, especially for mangaba tree botanical variety of Northeastern Brazil, limiting aspects such as ethylene accumulation in the cultivation flask and loss of vigor in subcultures have been observed. This study was aimed at assessing the technical and scientific knowledge of the in vitro propagation of botanical mangaba tree variety and at improving the micropropagation protocol, establishing the in vitro cultivation time, the best type of flask sealing and explant at different micropropagation stages. For the establishment phase and for the first and second subcultures, the MS medium with 3% sucrose and 0.6% agar, supplemented with 1 mg L-1 IAA and 1 mg L-1 BA was used. Evaluations were performed at 30, 50 and 65 days of in vitro cultivation. The best types of flask sealing for the establishment phase were the PVC film and Para-film® and for the first subculture the Para-film® seal. In the second subculture the PVC film and Para-film® seals promoted the best growth. The median and basal nodal segments presented the best performance in the first subculture. No significant effect of explant type was observed in the second subculture. The ideal subculture interval in the establishment phase and the first and second subcultures is 50 days.Na micropropagação, especialmente para mangaba, variedade botânica da árvore do Nordeste do Brasil, aspectos limitantes, como acúmulo de etileno no recipiente de cultivo e perda de vigor em subculturas têm sido observados. Neste estudo, objetivou-se avaliar o conhecimento técnico e científico da propagação in vitro de mangabeira, variedade botânica do Nordeste do Brasil, e melhorar o protocolo de micropropagação, o melhor tipo de vedação frasco e explante em diferentes etapas. Para a fase de estabelecimento e para as subculturas primeiro e segundo, foi utlizado o meio MS com 3% de sacarose e agar 0,6%, suplementado com 1 mg L-1 de AIA e 1 mg L-1 de BAP. As avaliações foram

  17. ERECTA-family receptor kinase genes redundantly prevent premature progression of secondary growth in the Arabidopsis hypocotyl.

    Science.gov (United States)

    Ikematsu, Shuka; Tasaka, Masao; Torii, Keiko U; Uchida, Naoyuki

    2017-03-01

    Secondary growth is driven by continuous cell proliferation and differentiation of the cambium that acts as vascular stem cells, producing xylem and phloem to expand vascular tissues laterally. During secondary growth of hypocotyls in Arabidopsis thaliana, the xylem undergoes a drastic phase transition from a parenchyma-producing phase to a fiber-producing phase at the appropriate time. However, it remains to be fully elucidated how progression of secondary growth is properly controlled. We focused on phenotypes of hypocotyl vasculatures caused by double mutation in ERECTA (ER) and ER-LIKE1 (ERL1) receptor-kinase genes to elucidate their roles in secondary growth. ER and ERL1 redundantly suppressed excessive radial growth of the hypocotyl vasculature during secondary growth. ER and ERL1 also prevented premature initiation of the fiber differentiation process mediated by the NAC SECONDARY WALL THICKENING PROMOTING FACTORs in the hypocotyl xylem. Upon floral transition, the hypocotyl xylem gained a competency to respond to GA in a BREVIPEDICELLUS-dependent manner, which was a prerequisite for fiber differentiation. However, even after the floral transition, ER and ERL1 prevented precocious initiation of the GA-mediated fiber formation. Collectively, our findings reveal that ER and ERL1 redundantly prevent premature progression of sequential events in secondary growth. © 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.

  18. Isolation and characterization of the betalain biosynthesis gene involved in hypocotyl pigmentation of the allotetraploid Chenopodium quinoa.

    Science.gov (United States)

    Imamura, Tomohiro; Takagi, Hiroki; Miyazato, Akio; Ohki, Shinya; Mizukoshi, Hiroharu; Mori, Masashi

    2018-02-05

    In quinoa seedlings, the pigment betalain accumulates in the hypocotyl. To isolate the genes involved in betalain biosynthesis in the hypocotyl, we performed ethyl methanesulfonate (EMS) mutagenesis on the CQ127 variety of quinoa seedlings. While putative amaranthin and celosianin II primarily accumulate in the hypocotyls, this process produced a green hypocotyl mutant (ghy). This MutMap+ method using the quinoa draft genome revealed that the causative gene of the mutant is CqCYP76AD1-1. Our results indicated that the expression of CqCYP76AD1-1 was light-dependent. In addition, the transient expression of CqCYP76AD1-1 in Nicotiana benthamiana leaves resulted in the accumulation of betanin but not isobetanin, and the presence of a polymorphism in CqCYP76A1-2 in the CQ127 variety was shown to have resulted in its loss of function. These findings suggested that CqCYP76AD1-1 is involved in betalain biosynthesis during the hypocotyl pigmentation process in quinoa. To our knowledge, CqCYP76AD1-1 is the first quinoa gene identified by EMS mutagenesis using a draft gene sequence. Copyright © 2018. Published by Elsevier Inc.

  19. Gravitropism in higher plant shoots. VI. Changing sensitivity to auxin in gravistimulated soybean hypocotyls

    Science.gov (United States)

    Rorabaugh, P. A.; Salisbury, F. B.

    1989-01-01

    Although the Cholodny-Went model of auxin redistribution has been used to explain the transduction phase of gravitropism for over 60 years, problems are apparent, especially with dicot stems. An alternative to an auxin gradient is a physiological gradient in which lower tissues of a horizontal stem become more sensitive than upper tissues to auxin already present. Changes in tissue sensitivity to auxin were tested by immersing marked Glycine max Merrill (soybean) hypocotyl sections in buffered auxin solutions (0, 10(-8) to 10(-2) molar indoleacetic acid) and observing bending and growth of upper and lower surfaces. The two surfaces of horizontal hypocotyl sections responded differently to the same applied auxin stimulus; hypocotyls bent up (lower half grew more) in buffer alone or in low auxin levels, but bent down (upper half grew more) in high auxin. Dose-response curves were evaluated with Michaelis-Menten kinetics, with auxin-receptor binding analogous to enzyme-substrate binding. Vmax for the lower half was usually greater than that for the upper half, which could indicate more binding sites in the lower half. Km of the upper half was always greater than that of the lower half (unmeasurably low), which could indicate that upper-half binding sites had a much lower affinity for auxin than lower-half sites. Dose-response curves were also obtained for sections scrubbed' (cuticle abraded) on top or bottom before immersion in auxin, and gravitropic memory' experiments of L. Brauner and A. Hagar (1958 Planta 51: 115-147) were duplicated. [1-14C]Indoleacetic acid penetration was equal into the two halves, and endogenous plus exogenously supplied (not radiolabeled) free auxin in the two halves (by gas chromatography-selected ion monitoring-mass spectrometry) was also equal. Thus, differential growth occurred without free auxin redistribution, contrary to Cholodny-Went but in agreement with a sensitivity model.

  20. Studying Secondary Growth and Bast Fiber Development: The Hemp Hypocotyl Peeks behind the Wall

    Czech Academy of Sciences Publication Activity Database

    Behr, M.; Legay, S.; Žižková, Eva; Motyka, Václav; Dobrev, Petre; Hausman, J.F.; Lutts, S.; Guerriero, G.

    2016-01-01

    Roč. 7, NOV 18 (2016), č. článku 1733. ISSN 1664-462X R&D Projects: GA ČR(CZ) GA16-14649S Institutional support: RVO:61389030 Keywords : plant-cell-wall * arabidopsis-thaliana * cannabis-sativa * lignin biosynthesis * microarray analysis * gene-expression * in-silico * anthocyanin biosynthesis * arabinogalactan-protein * transcription factors * Cannabis sativa * hypocotyl * bast fibers * cell wall * RNA-Seq * immunohistochemistry * phytohormones Subject RIV: EF - Botanics Impact factor: 4.298, year: 2016

  1. An efficient plant regeneration protocol from petiole explants of ...

    African Journals Online (AJOL)

    The highest percentage of shoot buds induction (64.0%) was observed on MS medium supplemented with 0.52 mgL-1 TDZ with organic additives; adenine sulphate (50 mgL-1) + glutamine (100 mgL-1) + L-arginine (25 mgL-1) + citric acid (0.0025%) + ascorbic acid (0.005%). A maximum of six shoots per explant were ...

  2. In vitro propagation of Alstroemeria using rhizome explants derived ...

    African Journals Online (AJOL)

    SERVER

    2007-09-19

    Sep 19, 2007 ... Single in vitro and in vivo rhizome bud were excised (3 -7 mm) using a sharp knife and cultured on MS basal medium with 3 different composition of growth regulators. Culture media. The explants were planted in culture media as M1 (MS + 1 mg l-1. BA + 0.2 mg l-1 NAA) Han et al. (1994), M2 (MS + 1 mg l-1 ...

  3. Restricted cell elongation in Arabidopsis hypocotyls is associated with a reduced average pectin esterification level

    Directory of Open Access Journals (Sweden)

    Derbyshire Paul

    2007-06-01

    Full Text Available Abstract Background Cell elongation is mainly limited by the extensibility of the cell wall. Dicotyledonous primary (growing cell walls contain cellulose, xyloglucan, pectin and proteins, but little is known about how each polymer class contributes to the cell wall mechanical properties that control extensibility. Results We present evidence that the degree of pectin methyl-esterification (DE% limits cell growth, and that a minimum level of about 60% DE is required for normal cell elongation in Arabidopsis hypocotyls. When the average DE% falls below this level, as in two gibberellic acid (GA mutants ga1-3 and gai, and plants expressing pectin methyl-esterase (PME1 from Aspergillus aculeatus, then hypocotyl elongation is reduced. Conclusion Low average levels of pectin DE% are associated with reduced cell elongation, implicating PMEs, the enzymes that regulate DE%, in the cell elongation process and in responses to GA. At high average DE% other components of the cell wall limit GA-induced growth.

  4. Microtubule bundling plays a role in ethylene-mediated cortical microtubule reorientation in etiolated Arabidopsis hypocotyls.

    Science.gov (United States)

    Ma, Qianqian; Sun, Jingbo; Mao, Tonglin

    2016-05-15

    The gaseous hormone ethylene is known to regulate plant growth under etiolated conditions (the 'triple response'). Although organization of cortical microtubules is essential for cell elongation, the underlying mechanisms that regulate microtubule organization by hormone signaling, including ethylene, are ambiguous. In the present study, we demonstrate that ethylene signaling participates in regulation of cortical microtubule reorientation. In particular, regulation of microtubule bundling is important for this process in etiolated hypocotyls. Time-lapse analysis indicated that selective stabilization of microtubule-bundling structures formed in various arrays is related to ethylene-mediated microtubule orientation. Bundling events and bundle growth lifetimes were significantly increased in oblique and longitudinal arrays, but decreased in transverse arrays in wild-type cells in response to ethylene. However, the effects of ethylene on microtubule bundling were partially suppressed in a microtubule-bundling protein WDL5 knockout mutant (wdl5-1). This study suggests that modulation of microtubule bundles that have formed in certain orientations plays a role in reorienting microtubule arrays in response to ethylene-mediated etiolated hypocotyl cell elongation. © 2016. Published by The Company of Biologists Ltd.

  5. Gene expression profile of zeitlupe/lov kelch protein1 T-DNA insertion mutants in Arabidopsis thaliana: Downregulation of auxin-inducible genes in hypocotyls.

    Science.gov (United States)

    Saitoh, Aya; Takase, Tomoyuki; Kitaki, Hiroyuki; Miyazaki, Yuji; Kiyosue, Tomohiro

    2015-01-01

    Elongation of hypocotyl cells has been studied as a model for elucidating the contribution of cellular expansion to plant organ growth. ZEITLUPE (ZTL) or LOV KELCH PROTEIN1 (LKP1) is a positive regulator of warmth-induced hypocotyl elongation under white light in Arabidopsis, although the molecular mechanisms by which it promotes hypocotyl cell elongation remain unknown. Microarray analysis showed that 134 genes were upregulated and 204 genes including 15 auxin-inducible genes were downregulated in the seedlings of 2 ztl T-DNA insertion mutants grown under warm conditions with continuous white light. Application of a polar auxin transport inhibitor, an auxin antagonist or an auxin biosynthesis inhibitor inhibited hypocotyl elongation of control seedlings to the level observed with the ztl mutant. Our data suggest the involvement of auxin and auxin-inducible genes in ZTL-mediated hypocotyl elongation.

  6. Enhanced micropropagation and tiller formation in sugarcane through pretreatment of explants with thidiazuron (TDZ).

    Science.gov (United States)

    Kumari, Kavita; Lal, Madan; Saxena, Sangeeta

    2017-10-01

    An efficient, simple and commercially applicable protocol for rapid micropropagation of sugarcane has been designed using variety Co 05011. Pretreatment of shoot tip explants with thidiazuron (TDZ) induced high frequency regeneration of shoot cultures with improved multiplication ratio. The highest frequency (80%) of shoot initiation in explants pretreated with 10 mg/l of TDZ was obtained during the study. Maximum 65% shoot cultures could be established from the explants pretreated with TDZ as compared to minimum 40% establishment in explants without pretreatment. The explants pretreated with 10 mg/l of TDZ required minimum 40 days for the establishment of shoot cultures as compared to untreated explants which required 60 days. The highest average number of shoots per culture (19.1) could be obtained from the explants pretreated with 10 mg/l of TDZ, indicating the highest multiplication ratio (1:6). Highest rooting (over 94%) was obtained in shoots regenerated from pretreated explants on ½ strength MS medium containing 5.0 mg/l of NAA and 50 g/l of sucrose within 15 days. Higher number of tillers/clump (15.3) could be counted in plants regenerated from pretreated explants than untreated ones (10.9 tillers/clump) in field condition, three months after transplantation. Molecular analysis using RAPD and DAMD markers suggested that the pretreatment of explants with TDZ did not adversely affect the genetic stability of regenerated plants and maintained high clonal purity.

  7. Mucus glycoprotein secretion by tracheal explants: effects of pollutants

    International Nuclear Information System (INIS)

    Last, J.A.; Kaizu, T.

    1980-01-01

    Tracheal slices incubated with radioactive precursors in tissue culture medium secrete labeled mucus glycoproteins into the culture medium. We have used an in vivtro approach, a combined method utilizing exposure to pneumotoxins in vivo coupled with quantitation of mucus secretion rates in vitro, to study the effects of inhaled pollutants on mucus biosynthesis by rat airways. In addition, we have purified the mucus glycoproteins secreted by rat tracheal explants in order to determine putative structural changes that might by the basis for the observed augmented secretion rates after exposure of rats to H2SO4 aerosols in combination with high ambient levels of ozone. After digestion with papain, mucus glycoproteins secreted by tracheal explants may be separated into five fractions by ion-exchange chromatography, with recovery in high yield, on columns of DEAE-cellulose. Each of these five fractions, one neutral and four acidic, migrates as a single unique spot upon cellulose acetate electrophoresis at pH values of 8.6 and 1.2. The neutral fraction, which is labeled with [3H] glucosamine, does not contain radioactivity when Na2 35SO4 is used as the precursor. Acidic fractions I to IV are all labeled with either 3H-glucosamine or Na2 35SO4 as precursor. Acidic fraction II contains sialic acid as the terminal sugar on its oligosaccharide side chains, based upon its chromatographic behavior on columns of wheat-germ agglutinin-Agarose. Treatment of this fraction with neuraminidase shifts its elution position in the gradient to a lower salt concentration, coincident with acidic fraction I. After removal of terminal sialic acid residues with either neuraminidase or low pH treatment, the resultant terminal sugar on the oligosaccharide side chains is fucose. These results are identical with those observed with mucus glycoproteins secreted by cultured human tracheal explants and purified by these same techniques

  8. Jasmonate inhibits COP1 activity to suppress hypocotyl elongation and promote cotyledon opening in etiolated Arabidopsis seedlings.

    Science.gov (United States)

    Zheng, Yuyu; Cui, Xuefei; Su, Liang; Fang, Shuang; Chu, Jinfang; Gong, Qingqiu; Yang, Jianping; Zhu, Ziqiang

    2017-06-01

    A germinating seedling undergoes skotomorphogenesis to emerge from the soil and reach for light. During this phase, the cotyledons are closed, and the hypocotyl elongates. Upon exposure to light, the seedling rapidly switches to photomorphogenesis by opening its cotyledons and suppressing hypocotyl elongation. The E3 ubiquitin ligase CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) is critical for maintaining skotomorphogenesis. Here, we report that jasmonate (JA) suppresses hypocotyl elongation and stimulates cotyledon opening in etiolated seedlings, partially phenocopying cop1 mutants in the dark. We also find that JA stabilizes several COP1-targeted transcription factors in a COP1-dependent manner. RNA-seq analysis further defines a JA-light co-modulated and cop1-dependent transcriptome, which is enriched for auxin-responsive genes and genes participating in cell wall modification. JA suppresses COP1 activity through at least two distinct mechanisms: decreasing COP1 protein accumulation in the nucleus; and reducing the physical interaction between COP1 and its activator, SUPPRESSOR OF PHYTOCHROME A-105 1 (SPA1). Our work reveals that JA suppresses COP1 activity to stabilize COP1 targets, thereby inhibiting hypocotyl elongation and stimulating cotyledon unfolding in etiolated Arabidopsis seedlings. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.

  9. Retardation of hypocotyl elongation of ornamental and vegetable seedlings by ultraviolet irradiation

    International Nuclear Information System (INIS)

    Bae, E.; Inamoto, K.; Doi, M.; Imanishi, H.

    1998-01-01

    Seedlings of cosmos (Cosmos bipinnatus Cav.), lettuce (Lactuca sativa L.), sunflower (Helianthus annuus L), ornamental kale (Brassica oleracea L. var. acephara), tomato (Lycopersicon esculentum Mill.), bell pepper (Capsicum annuum L.), and cucumber (Cucumis sativus L.) were irradiated by a UV-B lamp (fluorescent sun lamp) or a three-band fluorescent lamp (control) for 72 hr just after sowing. Hypocotyl elongation was repressed by all species during the irradiation with UV-B. The retarding effects of UV-B persisted when these seedlings were placed in dark or a greenhouse with 30% shade after irradiation. The most effective timing of UV-B irradiation for cosmos and ornamental kale seedlings was from 48 hr to 72 hr after sowing, when the seedlings were rapidly increasing their surface area to UV-B

  10. Proliferation of Female Inflorescences explants of Date Palm

    International Nuclear Information System (INIS)

    Sidky, R.A; Eldawyati, M.M

    2012-01-01

    This study was conducted to determine the effect of Abscisic acid (ABA) and Ancymidol on proliferation of female inflorescences explants of date palm. In the first experiment two lengths of spath at (5-7 cm) or at (7-10 cm) were cultured on nutrient media which consists of half macro and full micro salts of MS medium supplemented with gradual decreasing in concentration of Abscisic acid (ABA) and Ancymidol from 4.5, 3.0, 1.5 to 0.5 mg -1 . In the second experiment two phases of nutrient medium (solid and liquid) and two source of carbon were investigated. Gradual decreasing of ABA concentrations from 4.5 mg -1 to 1.5 mg -1 in culture medium, stimulated the production of direct somatic embryos and accelerated callus initiation, but at last decrement (0.5 mg -1 ) of Ancymidol concentration few embryos were produced. Callus initiation from inflorescences explants gave high production and well development of somatic embryos when cultured on liquid medium supplemented with 40 g -1 sucrose. All direct or indirect somatic embryos obtained in these experiments were converted successfully to healthy normal plantlets which could be transferred to acclimatization stage.

  11. Microscopic and spectroscopic investigation of an explanted opacified intraocular lens

    Energy Technology Data Exchange (ETDEWEB)

    Simon, V., E-mail: viosimon@phys.ubbcluj.ro [Babeş-Bolyai University, Faculty of Physics and Interdisciplinary Research Institute on Bio-Nano-Sciences, 400084 Cluj-Napoca (Romania); Radu, T.; Vulpoi, A. [Babeş-Bolyai University, Faculty of Physics and Interdisciplinary Research Institute on Bio-Nano-Sciences, 400084 Cluj-Napoca (Romania); Rosca, C. [Optilens Clinic of Ophthalmology, 400604 Cluj-Napoca (Romania); Eniu, D. [Iuliu Haţieganu University of Medicine and Pharmacy, Department of Molecular Sciences, 400349 Cluj-Napoca (Romania)

    2015-01-15

    Highlights: • Changes on intraocular lens (IOL) surface after implantation. • Partial opacification of IOL central area. • Elemental composition on IOL surface prior to and after implantation. • First XPS depth profiling examination of the opacifying deposits. • Cell-mediated hydroxyapatite structuring. - Abstract: The investigated polymethylmethacrylate intraocular lens explanted an year after implantation presented a fine granularity consisting of ring-like grains of about 15 μm in diameter. In order to evidence the changes occurred on intraocular lens relative to morphology, elemental composition and atomic environments, microscopic and spectroscopic analyses were carried out using scanning electron microscopy (SEM), Fourier transform infrared (FTIR), energy-dispersive X-ray (EDS), and X-ray photoelectron (XPS) spectroscopies. The results revealed that the grains contain hydroxyapatite mineral phase. A protein layer covers the lens both in opacified and transparent zones. The amide II band is like in basal epithelial cells. The shape and size of the grains, and the XPS depth profiling results indicate the possibility of a cell-mediated process involving lens epithelial cells which fagocitated apoptotic epithelial cells, and in which the debris derived from cell necrosis were calcified. To the best of our knowledge, this is the first investigation on explanted intraocular lenses using XPS depth profiling in order to examine the inside of the opacifying deposits.

  12. Active Segmentation.

    Science.gov (United States)

    Mishra, Ajay; Aloimonos, Yiannis

    2009-01-01

    The human visual system observes and understands a scene/image by making a series of fixations. Every fixation point lies inside a particular region of arbitrary shape and size in the scene which can either be an object or just a part of it. We define as a basic segmentation problem the task of segmenting that region containing the fixation point. Segmenting the region containing the fixation is equivalent to finding the enclosing contour- a connected set of boundary edge fragments in the edge map of the scene - around the fixation. This enclosing contour should be a depth boundary.We present here a novel algorithm that finds this bounding contour and achieves the segmentation of one object, given the fixation. The proposed segmentation framework combines monocular cues (color/intensity/texture) with stereo and/or motion, in a cue independent manner. The semantic robots of the immediate future will be able to use this algorithm to automatically find objects in any environment. The capability of automatically segmenting objects in their visual field can bring the visual processing to the next level. Our approach is different from current approaches. While existing work attempts to segment the whole scene at once into many areas, we segment only one image region, specifically the one containing the fixation point. Experiments with real imagery collected by our active robot and from the known databases 1 demonstrate the promise of the approach.

  13. Effect of Macrophage Migration Inhibitory Factor (MIF) in Human Placental Explants Infected with Toxoplasma gondii Depends on Gestational Age

    Science.gov (United States)

    de Oliveira Gomes, Angelica; de Oliveira Silva, Deise Aparecida; Silva, Neide Maria; de Freitas Barbosa, Bellisa; Franco, Priscila Silva; Angeloni, Mariana Bodini; Fermino, Marise Lopes; Roque-Barreira, Maria Cristina; Bechi, Nicoletta; Paulesu, Luana Ricci; dos Santos, Maria Célia; Mineo, José Roberto; Ferro, Eloisa Amália Vieira

    2011-01-01

    Because macrophage migration inhibitory factor (MIF) is a key cytokine in pregnancy and has a role in inflammatory response and pathogen defense, the objective of the present study was to investigate the effects of MIF in first- and third-trimester human placental explants infected with Toxoplasma gondii. Explants were treated with recombinant MIF, IL-12, interferon-γ, transforming growth factor-β1, or IL-10, followed by infection with T. gondii RH strain tachyzoites. Supernatants of cultured explants were assessed for MIF production. Explants were processed for morphologic analysis, immunohistochemistry, and real-time PCR analysis. Comparison of infected and stimulated explants versus noninfected control explants demonstrated a significant increase in MIF release in first-trimester but not third-trimester explants. Tissue parasitism was higher in third- than in first-trimester explants. Moreover, T. gondii DNA content was lower in first-trimester explants treated with MIF compared with untreated explants. However, in third-trimester explants, MIF stimulus decreased T. gondii DNA content only at the highest concentration of the cytokine. In addition, high expression of MIF receptor was observed in first-trimester placental explants, whereas MIF receptor expression was low in third-trimester explants. In conclusion, MIF was up-regulated and demonstrated to be important for control of T. gondii infection in first-trimester explants, whereas lack of MIF up-regulation in third-trimester placentas may be involved in higher susceptibility to infection at this gestational age. PMID:21641401

  14. Utilization of Aseptic Seedling Explants for In vitro Propagation of Indian Red Wood

    Directory of Open Access Journals (Sweden)

    Kishore Kumar CHIRUVELLA

    2013-12-01

    Full Text Available Micropropagation has been advocated as one of the most viable biotechnological tool for ex situ conservation of rare, endangered endemic medicinal plants germplasm. Rapid clonal micropropagation protocol for large-scale multiplication of an endemic medicinal plant Soymida febrifuga (Meliaceae was established from 15-day aseptic seedling cotyledonary node and shoot tip explants. High frequency of sprouting and shoot differentiation was observed from cotyledonary node explants compared to shoot tip, on Murashige and Skoog (MS medium fortified with BA, KN, 2-iP and CM. Of the cytokinins used, BA (3.0 mgl-1 supported highest average number and maximum multiple shoot differentiation (16.6. In vitro proliferated shoots were multiplied rapidly by culturing nodal segments as microcuttings, further subcultured on the same media for elongation. Elongated shoots upon transfer to MS medium fortified with IBA showed rooting within two weeks of culture. Rooted plantlets were successfully hardened and 75% of rooted shoots successfully survived on establishment to the soil. Plants looked healthy with no visually detectable phenotypic variations. This protocol provides a successful and rapid technique that can be used for ex situ conservation minimizing the pressure on wild populations and contributes to the conservation of this endemic medicinally potent flora.

  15. In Vitro Regeneration of Shoots From Nodal Explants of Dendrobium Chrysotoxum Lindl

    Directory of Open Access Journals (Sweden)

    Kaur Saranjeet

    2017-06-01

    Full Text Available Transverse sections (2 mm thickness of stem-nodes from in vitro raised seedlings had morphogenic potential on semisolid and liquid Murashige and Skoog medium supplemented with cytokinins N6-benzyladenine (BA 4.44 μM, furfurylaminopurine (KIN 4.65 μM and auxin α-naphthalene acetic acid (NAA 5.37 μM individually and in combinations. The regeneration response was influenced by both the type of growth regulator and physical state of the medium. The explants produced either shoot buds on cytokinincontaining media or protocorm-like bodies (PLBs on NAA containing media both solid and liquid. More neo-formations were produced on liquid media, especially those containing only NAA. They were formed at nodal and inter-nodal regions. The secondary buds were produced on the surface of primary PLBs. The plantlets were developed on MS medium containing banana homogenate 50 g·dm-3. The current study is the first ever report on successful regeneration of Dendrobium chrysotoxum from stem-node segments.

  16. Segmentation: Identification of consumer segments

    DEFF Research Database (Denmark)

    Høg, Esben

    2005-01-01

    It is very common to categorise people, especially in the advertising business. Also traditional marketing theory has taken in consumer segments as a favorite topic. Segmentation is closely related to the broader concept of classification. From a historical point of view, classification has its...... origin in other sciences as for example biology, anthropology etc. From an economic point of view, it is called segmentation when specific scientific techniques are used to classify consumers to different characteristic groupings. What is the purpose of segmentation? For example, to be able to obtain...... a basic understanding of grouping people. Advertising agencies may use segmentation totarget advertisements, while food companies may usesegmentation to develop products to various groups of consumers. MAPP has for example investigated the positioning of fish in relation to other food products...

  17. Multiplicação in vitro do porta-enxerto de macieira cv. Marubakaido: efeito da orientação do explante no meio de cultura In vitro multiplication of the apple rootstock cv. Marubakaido: effect of the orientation of explant in the medium of culture

    Directory of Open Access Journals (Sweden)

    ALAN CRISTIANO ERIG

    2002-08-01

    Full Text Available Objetivou-se avaliar o efeito da orientação do explante, vertical ou horizontal, no meio de cultura, na multiplicação in vitro, do porta-enxerto de macieira cv. Marubakaido. O meio de cultura utilizado foi o MS com N (nitrogênio reduzido a ¾ da concentração original, 100mg.L-1 de mio-inositol, 40g.L-1 de sacarose e 6g.L-1 de ágar, suplementado com 4,44mM de BAP (6-benzilaminopurina e 0,2ml.L-1 de PPM TM ("Plant Preservative Mixture". Segmentos caulinares com duas gemas e o ápice excisado foram utilizados como explantes. Após a inoculação, os frascos com os explantes foram incubados a 16 horas de fotoperíodo, à temperatura de 25±2ºC, com radiação de 25µmoles.m-2.s-1. O número de brotações, o número de gemas por explante, a taxa de multiplicação e a altura da brotação maior foram avaliados aos quarenta dias de cultivo. O maior número de brotações, o maior número de gemas e a maior taxa de multiplicação foram obtidos com o explante na orientação horizontal no meio de cultura. Não houve diferença significativa quanto à orientação vertical e horizontal do explante no meio de cultura para a altura da brotação maior.The aim of this study was evaluate the effect of the vertical and the horizontal orientation of the explant in the culture medium, in the in vitro multiplication, for the apple rootstock cv. Marubakaido. The culture medium used was the MS with N reduced to ¾ of the original concentration, myo-inositol (100mg.L-1, sucrose (40g.L-1 and agar (6g.L-1, suplemented with BAP (4.44mM and PPM TM (0.2ml.L-1. Stem segments with two buds and the apex excised were used as explants. After the inoculation, the flasks with the explants were incubated at 16 hour of photoperiod, 25±2ºC temperature, with irradiation of 25µmoles.m-2.s-1. The number of shoots and buds, the rate of multiplication and the height of the larger shoot were evaluated after 40 days of cultivation. The highests shoot number, number of buds

  18. Segmental Vitiligo.

    Science.gov (United States)

    van Geel, Nanja; Speeckaert, Reinhart

    2017-04-01

    Segmental vitiligo is characterized by its early onset, rapid stabilization, and unilateral distribution. Recent evidence suggests that segmental and nonsegmental vitiligo could represent variants of the same disease spectrum. Observational studies with respect to its distribution pattern point to a possible role of cutaneous mosaicism, whereas the original stated dermatomal distribution seems to be a misnomer. Although the exact pathogenic mechanism behind the melanocyte destruction is still unknown, increasing evidence has been published on the autoimmune/inflammatory theory of segmental vitiligo. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Articular cartilage explant culture; an appropriate in vitro system to compare osteoarthritic and normal human cartilage

    NARCIS (Netherlands)

    Lafeber, F. P.; Vander Kraan, P. M.; van Roy, J. L.; Huber-Bruning, O.; Bijlsma, J. W.

    1993-01-01

    Proteoglycan metabolism of normal and histologically mild to moderate osteoarthritic cartilage explants were studied. Explants were obtained from the human knee of donors aged over 40 years. Proteoglycan content, synthesis and release were very similar in normal cartilage obtained from donors with

  20. EXPLANTATION OF MESANGIAL CELL HILLOCKS - A METHOD FOR OBTAINING HUMAN MESANGIAL CELLS IN CULTURE

    NARCIS (Netherlands)

    MULLER, EW; KIM, Y; MICHAEL, AF; VERNIER, RL; VANDERHEM, GK; VANDERWOUDE, FJ

    A simple method is presented for selective cell culture of human mesangial cells using explanatation of mesangial cell hillocks. Glomeruli which had been incubated with collagenase were explanted on plastic tissue culture flasks. Three to 6 weeks after explantation, a rapidly growing multilayer of

  1. Hypoxia inhibits hypertrophic differentiation and endochondral ossification in explanted tibiae.

    Directory of Open Access Journals (Sweden)

    Jeroen C H Leijten

    Full Text Available Hypertrophic differentiation of growth plate chondrocytes induces angiogenesis which alleviates hypoxia normally present in cartilage. In the current study, we aim to determine whether alleviation of hypoxia is merely a downstream effect of hypertrophic differentiation as previously described or whether alleviation of hypoxia and consequent changes in oxygen tension mediated signaling events also plays an active role in regulating the hypertrophic differentiation process itself.Fetal mouse tibiae (E17.5 explants were cultured up to 21 days under normoxic or hypoxic conditions (21% and 2.5% oxygen respectively. Tibiae were analyzed on growth kinetics, histology, gene expression and protein secretion.The oxygen level had a strong influence on the development of explanted fetal tibiae. Compared to hypoxia, normoxia increased the length of the tibiae, length of the hypertrophic zone, calcification of the cartilage and mRNA levels of hypertrophic differentiation-related genes e.g. MMP9, MMP13, RUNX2, COL10A1 and ALPL. Compared to normoxia, hypoxia increased the size of the cartilaginous epiphysis, length of the resting zone, calcification of the bone and mRNA levels of hyaline cartilage-related genes e.g. ACAN, COL2A1 and SOX9. Additionally, hypoxia enhanced the mRNA and protein expression of the secreted articular cartilage markers GREM1, FRZB and DKK1, which are able to inhibit hypertrophic differentiation.Collectively our data suggests that oxygen levels play an active role in the regulation of hypertrophic differentiation of hyaline chondrocytes. Normoxia stimulates hypertrophic differentiation evidenced by the expression of hypertrophic differentiation related genes. In contrast, hypoxia suppresses hypertrophic differentiation of chondrocytes, which might be at least partially explained by the induction of GREM1, FRZB and DKK1 expression.

  2. Morphologic differences observed by scanning electron microscopy according to the reason for pseudophakic IOL explantation

    DEFF Research Database (Denmark)

    Fernandez-Buenaga, Roberto; Alio, Jorge L.; Ramirez, Jose M.

    2015-01-01

    Purpose To compare variations in surface morphology, as studied by scanning electron microscopy (SEM), of explanted intraocular lenses (IOLs) concerning the cause leading to the explantation surgery. Methods In this prospective multicenter study, explanted IOLs were analyzed by SEM and energy...... explanted due to dislocation demonstrated calcifications in 8 lenses (50%), salt precipitates in 6 cases (37.5%), and erythrocytes and fibrosis/fibroblasts in 2 cases (12.5%). In the refractive error cases, the SEM showed proteins in 5 cases (45.5%) and salt precipitates in 4 lenses (36.4%). In IOL...... opacification, the findings were calcifications in 2 of the 3 lenses (66.6%) and proteins in 2 lenses (66.6%). Conclusions A marked variation in surface changes was observed by SEM. Findings did not correlate with cause for explantation. Scanning electron microscopy is a useful tool that provides exclusive...

  3. A simple technique of intraocular lenses explantation for single-piece foldable lenses

    Directory of Open Access Journals (Sweden)

    Arup Bhaumik

    2017-01-01

    Full Text Available Foldable intraocular lenses (IOLs are most commonly used in modern-day cataract surgery. Explantation of these IOLs is not frequently encountered, but sometimes extreme situations may demand the same. Commonly explantation is achieved by bisecting the IOL inside the anterior chamber with a cutter and delivering the pieces out one by one. This may require corneal wound extension with associated damage and endothelial loss leading to visual deterioration. We devised a simple, innovative IOL explantation technique utilizing a modified Alcon A cartridge and snare. This can successfully refold the IOL to be explanted inside the eye and deliver it out through the same wound. The device has limitations with very thick optic lenses, multipiece, and silicon IOLs. In conclusion, we describe a simple, innovative, and reproducible technique to explant almost any single piece IOL without compromising the original surgery and yielding very satisfactory outcomes.

  4. In vitro plant regeneration of Albizia lebbeck (L. from seed explants

    Directory of Open Access Journals (Sweden)

    S. Perveen

    2013-07-01

    Full Text Available Objectives: An efficient and reproducible regeneration protocol for rapid multiplication of Albizia lebbeck (L. was developed by using intact seed explants.Methods: Murashige and Skoog's (MS medium supplemented with different hormones (BA, Kn, GA3 and TDZ was used for the induction of multiple shoots from the seed explants. Ex-vitro rooting was performed by using pulse treatment method in auxins (IBA and NAA and the complete plantlets were transferred to the field.Results: High frequency direct shoot induction was found in aseptic seed cultures of A. lebbeck on Murashige and Skoog medium supplemented with 5.0 µM TDZ (Thiadiazuron. Seeds were germinated after 7 days of culture and induced maximum 8 shoots from the region adjacent to the apex of the primary shoot of the seedling upto 25 days of incubation. Proliferating shoot cultures with increased shoot length was established by sub-culture of excised sprouting epicotyls on MS medium supplied with reduced concentrations of TDZ. Maximum shoot regeneration frequency (76 % with  highest number of shoots (21 and shoot length (5.1 cm per sprouting epicotyl was observed in the MS medium supplemented with 0.5 µM TDZ after 8 weeks of culture. Different concentrations of Indole-3-butyric acid (IBA and α-naphthalene acetic acid (NAA were tested to determine the optimal conditions for ex-vitro rooting of the microshoots. The best treatment for maximum ex-vitro root induction frequency (81 % was accomplished with IBA (250 µM pulse treatment given to the basal end of the microshoots for 30 min followed by their transfer in plastic cups containing soilrite and eventually established in normal garden soil + soilrite (1:1 with 78 % survival rate. In addition, histological study was undertaken to gain a better understanding of the regenerated shoots from the epicotyl region.Conclusion: The findings will be fruitful in getting a time saving and cost effective protocol for the in vitro propagation of Albizia

  5. A Study of Gibberellin Homeostasis and Cryptochrome-Mediated Blue Light Inhibition of Hypocotyl Elongation1[W][OA

    Science.gov (United States)

    Zhao, Xiaoying; Yu, Xuhong; Foo, Eloise; Symons, Gregory M.; Lopez, Javier; Bendehakkalu, Krishnaprasad T.; Xiang, Jing; Weller, James L.; Liu, Xuanming; Reid, James B.; Lin, Chentao

    2007-01-01

    Cryptochromes mediate blue light-dependent photomorphogenic responses, such as inhibition of hypocotyl elongation. To investigate the underlying mechanism, we analyzed a genetic suppressor, scc7-D (suppressors of cry1cry2), which suppressed the long-hypocotyl phenotype of the cry1cry2 (cryptochrome1/cryptochrome2) mutant in a light-dependent but wavelength-independent manner. scc7-D is a gain-of-expression allele of the GA2ox8 gene encoding a gibberellin (GA)-inactivating enzyme, GA 2-oxidase. Although scc7-D is hypersensitive to light, transgenic seedlings expressing GA2ox at a level higher than scc7-D showed a constitutive photomorphogenic phenotype, confirming a general role of GA2ox and GA in the suppression of hypocotyl elongation. Prompted by this result, we investigated blue light regulation of mRNA expression of the GA metabolic and catabolic genes. We demonstrated that cryptochromes are required for the blue light regulation of GA2ox1, GA20ox1, and GA3ox1 expression in transient induction, continuous illumination, and photoperiodic conditions. The kinetics of cryptochrome induction of GA2ox1 expression and cryptochrome suppression of GA20ox1 or GA3ox1 expression correlate with the cryptochrome-dependent transient reduction of GA4 in etiolated wild-type seedlings exposed to blue light. Therefore we propose that in deetiolating seedlings, cryptochromes mediate blue light regulation of GA catabolic/metabolic genes, which affect GA levels and hypocotyl elongation. Surprisingly, no significant change in the GA4 content was detected in the whole shoot samples of the wild-type or cry1cry2 seedlings grown in the dark or continuous blue light, suggesting that cryptochromes may also regulate GA responsiveness and/or trigger cell- or tissue-specific changes of the level of bioactive GAs. PMID:17644628

  6. Roles of Proteome Dynamics and Cytokinin Signaling in Root to Hypocotyl Ratio Changes Induced by Shading Roots of Arabidopsis Seedlings

    Czech Academy of Sciences Publication Activity Database

    Novak, J.; Černý, M.; Pavlů, J.; Zemánková, J.; Skalák, J.; Plačková, Lenka; Brzobohatý, Břetislav

    2015-01-01

    Roč. 56, č. 5 (2015), s. 1006-1018 ISSN 0032-0781 R&D Projects: GA MŠk(CZ) LO1204 Grant - others:GA ČR(CZ) GAP305/12/2144 Institutional support: RVO:68081707 ; RVO:61389030 Keywords : Arabidopsis thaliana * Cytokinin * Hypocotyl elongation Subject RIV: BO - Biophysics; EF - Botanics (UEB-Q) Impact factor: 4.319, year: 2015

  7. Over-expression of mango (Mangifera indica L.) MiARF2 inhibits root and hypocotyl growth of Arabidopsis.

    Science.gov (United States)

    Wu, Bei; Li, Yun-He; Wu, Jian-Yong; Chen, Qi-Zhu; Huang, Xia; Chen, Yun-Feng; Huang, Xue-Lin

    2011-06-01

    An auxin response factor 2 gene, MiARF2, was cloned in our previous study [1] from the cotyledon section of mango (Mangifera indica L. cv. Zihua) during adventitious root formation, which shares an 84% amino acid sequence similarity to Arabidopsis ARF2. This study was to examine the effects of over-expression of the full-length MiARF2 open reading frame on the root and hypocotyl growth in Arabidopsis. Phenotype analysis showed that the T(3) transgenic lines had about 20-30% reduction in the length of hypocotyls and roots of the seedlings in comparison with the wild-type. The transcription levels of ANT and ARGOS genes which play a role in controlling organ size and cell proliferation in the transgenic seedlings also decreased. Therefore, the inhibited root and hypocotyl growth in the transgenic seedlings may be associated with the down-regulated transcription of ANT and ARGOS by the over-expression of MiARF2. This study also suggests that although MiARF2 only has a single DNA-binding domain (DBD), it can function as other ARF-like proteins containing complete DBD, middle region (MR) and carboxy-terminal dimerization domain (CTD).

  8. Cell number, tissue thickness and protein content as measures for development and variability in cultured neocortex explants

    NARCIS (Netherlands)

    de Jong, B. M.; Ruijter, J. M.

    1989-01-01

    The development of neuronal number, explant thickness and amount of protein was studied in several series of rat neocortex explants, cultured up to 21 days in vitro (DIV). In contrast to the dimensions of the explant, which rapidly stabilized, the amount of protein showed a prolonged increase with

  9. A near-null magnetic field affects cryptochrome-related hypocotyl growth and flowering in Arabidopsis

    Science.gov (United States)

    Xu, Chunxiao; Yin, Xiao; Lv, Yan; Wu, Changzhe; Zhang, Yuxia; Song, Tao

    2012-03-01

    The blue light receptor, cryptochrome, has been suggested to act as a magnetoreceptor based on the proposition that photochemical reactions are involved in sensing the geomagnetic field. But the effects of the geomagnetic field on cryptochrome remain unclear. Although the functions of cryptochrome have been well demonstrated for Arabidopsis, the effect of the geomagnetic field on the growth of Arabidopsis and its mechanism of action are poorly understood. We eliminated the local geomagnetic field to grow Arabidopsis in a near-null magnetic field and found that the inhibition of Arabidopsis hypocotyl growth by white light was weakened, and flowering time was delayed. The expressions of three cryptochrome-signaling-related genes, PHYB, CO and FT also changed; the transcript level of PHYB was elevated ca. 40%, and that of CO and FT was reduced ca. 40% and 50%, respectively. These data suggest that the effects of a near-null magnetic field on Arabidopsis are cryptochrome-related, which may be revealed by a modification of the active state of cryptochrome and the subsequent signaling cascade.

  10. Biochemical characterization of the plasma membrane H+ - ATPase from red beet (Beta vulgaris) hypocotyl tissue

    International Nuclear Information System (INIS)

    Oleski, N.A.

    1986-01-01

    Several biochemical techniques including selective solubilization followed by gel filtration or various types of affinity chromatography, and antibody production were employed in an attempt to purify the plasma membrane H + - ATPase from red beet hypocotyl tissue. While the enzyme could not be purified using any of these methods, it was possible to successfully conduct a more detailed biochemical analysis of the H + - ATPase. The molecular weight and isoelectric point of the enzyme were determined using N,N'dicyclohexylcarbodiimide (DCCD) and a H + - ATPase antibody. When plasma membrane vesicles were incubated with 20 μM [ 14 C]-DCCD at 0 C, a single 97,000 dalton protein was apparent on a fluorograph. A close correlation between [ 14 C]-DCCD labelling of the 97,000 dalton protein and the extent of ATPase inhibition over a range of DCCD concentrations suggests that this 97,000 dalton protein is a component of the plasma membrane H + - ATPase. An antibody raised against the plasma membrane H + - ATPase of Neurospora crassa cross-reacted with the 97,000 dalton DCCD-binding protein, further supporting the identity of this protein. Immunoblots of two dimensional gels of red beet plasma membrane vesicles indicated the isoelectric point of the enzyme to be pH 6.5

  11. Optic nerve compression as a late complication of a hydrogel explant with silicone encircling band

    Directory of Open Access Journals (Sweden)

    Niels Crama

    2018-06-01

    Full Text Available Purpose: To present a complication of compressive optic neuropathy caused by a swollen hydrogel explant and posteriorly displaced silicone encircling band. Observations: A 72-year-old female patient presented with progressive visual loss and a tilted optic disc. Her medical history included a retinal detachment in 1993 that was treated with a hydrogel explant under a solid silicone encircling band. Visual acuity had decreased from 6/10 to 6/20 and perimetry showed a scotoma in the temporal superior quadrant. On Magnetic Resonance Imaging (MRI, compression of the optic nerve by a displaced silicone encircling band inferior nasally in combination with a swollen episcleral hydrogel explant was observed. Surgical removal of the hydrogel explant and silicone encircling band was uneventful and resulted in improvement of visual acuity and visual field loss. Conclusions and importance: This is the first report on compressive optic neuropathy caused by swelling of a hydrogel explant resulting in a dislocated silicone encircling band. The loss of visual function resolved upon removal of the explant and encircling band. Keywords: Retinal detachment, Tilted disc, Optic neuropathy, Miragel, Explant, Encircling band

  12. The Effects of Polyphenol Oxidase and Cycloheximide on the Early Stage of Browning in Phalaenopsis Explants

    Directory of Open Access Journals (Sweden)

    Xu Chuanjun

    2015-11-01

    Full Text Available Explant browning is one of the major problems in the tissue culture process, and polyphenol oxidase (PPO, is the major proteases involved in plant tissue browning. We investigated the effects of polyphenol oxidase on the early stage of browning in explants of the orchid Phalaenopsis. Our results show that PPO activity was significantly higher in explants cultured for 3 d than in the 0 h control. The levels of PPO transcripts and PPO protein were significantly higher in explants cultured for 6 h compared to the 0 h control; these high expression levels were maintained over increasing cultivation time. Cycloheximide (CHX treatment reduced PPO transcript levels, PPO protein levels, and PPO enzyme activity. High levels of PPO mRNA and PPO protein were detected in the cytoplasm and vascular bundles of Phalaenopsis explants cultured for 6 h compared to explants cultured for 0 h, 24 h, and 3 d. CHX treatment did not significantly affect the distribution of PPO mRNA and PPO protein in explant tissues, but their levels were significantly lower than those of the untreated control.

  13. The metabolic dynamics of cartilage explants over a long-term culture period

    Directory of Open Access Journals (Sweden)

    E.K Moo

    2011-01-01

    Full Text Available INTRODUCTION: Although previous studies have been performed on cartilage explant cultures, the generalized dynamics of cartilage metabolism after extraction from the host are still poorly understood due to differences in the experimental setups across studies, which in turn prevent building a complete picture. METHODS: In this study, we investigated the response of cartilage to the trauma sustained during extraction and determined the time needed for the cartilage to stabilize. Explants were extracted aseptically from bovine metacarpal-phalangeal joints and cultured for up to 17 days. RESULTS: The cell viability, cell number, proteoglycan content, and collagen content of the harvested explants were analyzed at 0, 2, 10, and 17 days after explantation. A high percentage of the cartilage explants were found to be viable. The cell density initially increased significantly but stabilized after two days. The proteoglycan content decreased gradually over time, but it did not decrease to a significant level due to leakage through the distorted peripheral collagen network and into the bathing medium. The collagen content remained stable for most of the culture period until it dropped abruptly on day 17. CONCLUSION: Overall, the tested cartilage explants were sustainable over long-term culture. They were most stable from day 2 to day 10. The degradation of the collagen on day 17 did not reach diseased levels, but it indicated the potential of the cultures to develop into degenerated cartilage. These findings have implications for the application of cartilage explants in pathophysiological fields.

  14. Reproducible simulation of respiratory motion in porcine lung explants

    Energy Technology Data Exchange (ETDEWEB)

    Biederer, J. [Dept. of Diagnostic Radiology, Univ. Hospital Schleswig-Holstein, Campus Kiel (Germany); Dept. of Radiology, German Cancer Research Center, Heidelberg (Germany); Plathow, C. [Dept. of Diagnostic Radiology, Eberhard-Karls-Univ. Tuebingen (Germany); Dept. of Radiology, German Cancer Research Center, Heidelberg (Germany); Schoebinger, M.; Meinzer, H.P. [Dept. of Medical and Biological Informatics, German Cancer Research Center, Heidelberg (Germany); Tetzlaff, R.; Puderbach, M.; Zaporozhan, J.; Kauczor, H.U. [Dept. of Radiology, German Cancer Research Center, Heidelberg (Germany); Bolte, H.; Heller, M. [Dept. of Diagnostic Radiology, Univ. Hospital Schleswig-Holstein, Campus Kiel (Germany)

    2006-11-15

    Purpose: To develop a model for exactly reproducible respiration motion simulations of animal lung explants inside an MR-compatible chest phantom. Materials and Methods: The materials included a piston pump and a flexible silicone reconstruction of a porcine diaphragm and were used in combination with an established MR-compatible chest phantom for porcine heart-lung preparations. The rhythmic inflation and deflation of the diaphragm at the bottom of the artificial thorax with water (1-1.5 L) induced lung tissue displacement resembling diaphragmatic breathing. This system was tested on five porcine heart-lung preparations using 1.5T MRI with transverse and coronal 3D-GRE (TR/TE=3.63/1.58, 256 x 256 matrix, 350 mm FOV, 4 mm slices) and half Fourier T2-FSE (TR/TE=545/29, 256 x 192, 350 mm, 6 mm) as well as multiple row detector CT (16 x 1 mm collimation, pitch 1.5, FOV 400 mm, 120 mAs) acquired at five fixed inspiration levels. Dynamic CT scans and coronal MRI with dynamic 2D-GRE and 2D-SS-GRE sequences (image frequencies of 10/sec and 3/sec, respectively) were acquired during continuous 'breathing' (7/minute). The position of the piston pump was visually correlated with the respiratory motion visible through the transparent wall of the phantom and with dynamic displays of CT and MR images. An elastic body splines analysis of the respiratory motion was performed using CT data. Results: Visual evaluation of MRI and CT showed three-dimensional movement of the lung tissue throughout the respiration cycle. Local tissue displacement inside the lung explants was documented with motion maps calculated from CT. The maximum displacement at the top of the diaphragm (mean 26.26 [SD 1.9] mm on CT and 27.16 [SD 1.5] mm on MRI, respectively [p=0.25; Wilcoxon test]) was in the range of tidal breathing in human patients. Conclusion: The chest phantom with a diaphragmatic pump is a promising platform for multi-modality imaging studies of the effects of respiratory lung

  15. Reproducible simulation of respiratory motion in porcine lung explants

    International Nuclear Information System (INIS)

    Biederer, J.; Plathow, C.; Schoebinger, M.; Meinzer, H.P.; Tetzlaff, R.; Puderbach, M.; Zaporozhan, J.; Kauczor, H.U.; Bolte, H.; Heller, M.

    2006-01-01

    Purpose: To develop a model for exactly reproducible respiration motion simulations of animal lung explants inside an MR-compatible chest phantom. Materials and Methods: The materials included a piston pump and a flexible silicone reconstruction of a porcine diaphragm and were used in combination with an established MR-compatible chest phantom for porcine heart-lung preparations. The rhythmic inflation and deflation of the diaphragm at the bottom of the artificial thorax with water (1-1.5 L) induced lung tissue displacement resembling diaphragmatic breathing. This system was tested on five porcine heart-lung preparations using 1.5T MRI with transverse and coronal 3D-GRE (TR/TE=3.63/1.58, 256 x 256 matrix, 350 mm FOV, 4 mm slices) and half Fourier T2-FSE (TR/TE=545/29, 256 x 192, 350 mm, 6 mm) as well as multiple row detector CT (16 x 1 mm collimation, pitch 1.5, FOV 400 mm, 120 mAs) acquired at five fixed inspiration levels. Dynamic CT scans and coronal MRI with dynamic 2D-GRE and 2D-SS-GRE sequences (image frequencies of 10/sec and 3/sec, respectively) were acquired during continuous 'breathing' (7/minute). The position of the piston pump was visually correlated with the respiratory motion visible through the transparent wall of the phantom and with dynamic displays of CT and MR images. An elastic body splines analysis of the respiratory motion was performed using CT data. Results: Visual evaluation of MRI and CT showed three-dimensional movement of the lung tissue throughout the respiration cycle. Local tissue displacement inside the lung explants was documented with motion maps calculated from CT. The maximum displacement at the top of the diaphragm (mean 26.26 [SD 1.9] mm on CT and 27.16 [SD 1.5] mm on MRI, respectively [p=0.25; Wilcoxon test]) was in the range of tidal breathing in human patients. Conclusion: The chest phantom with a diaphragmatic pump is a promising platform for multi-modality imaging studies of the effects of respiratory lung motion. (orig.)

  16. Mixed segmentation

    DEFF Research Database (Denmark)

    Hansen, Allan Grutt; Bonde, Anders; Aagaard, Morten

    content analysis and audience segmentation in a single-source perspective. The aim is to explain and understand target groups in relation to, on the one hand, emotional response to commercials or other forms of audio-visual communication and, on the other hand, living preferences and personality traits...

  17. Direct organogenesis of seaside heliotrope (Heliotropium crassavicum) using stem explants.

    Science.gov (United States)

    Satyavani, K; Dheepak, V; Gurudeeban, S; Ramanathan, T

    2013-10-15

    Heliotropium crassavicum L. is a sand binder salt marsh herb with enormous traditional value and widely found in South Asia America and Europe. In the direct method of regeneration from stem explants, we observed the maximum number of shoot regeneration after four weeks culture of MS elongation medium with 2.0 mg L(-1) of 2, 4-D (17.27 +/- 0.51). It was clear that MS medium with 2.0 mg mL(-1) 2, 4-D alone suitable for shoot multiplication as well as shoot elongation then compared to other combination of auxin and cytokinin. In vitro shoots were excised from shoot clumps and transferred to rooting medium containing 2, 4-dichlorophenoxy acetic acid (0.5-3.0 mg L(-1)). The maximum number of root regeneration (6.4 +/- 0.416) and root length (6.08 +/- 0.07) were observed in MS rooting medium fortified with 2.5 mg L(-1) of 2, 4-D after 2 weeks of culture. 85% of in vitro raised plantlets with well-developed shoots and roots were transferred to ex vivo conditions into polythene bag containing sterile compost with ratio (v/v/v) of organic fertilizer: sand: peat (1:2:2; 3:1:0 or 2:2:1). Sixty five percent of acclimated plants were transferred to the pots under full sun where they grew well without any detectable phenotypic variations.

  18. Space-time analysis of gravitropism in etiolated Arabidopsis hypocotyls using bioluminescence imaging of the IAA19 promoter fusion with a destabilized luciferase reporter.

    Science.gov (United States)

    Yamamoto, Kotaro T; Watahiki, Masaaki K; Matsuzaki, Jun; Satoh, Soichirou; Shimizu, Hisayo

    2017-07-01

    Imaging analysis was carried out during the gravitropic response of etiolated Arabidopsis hypocotyls, using an IAA19 promoter fusion of destabilized luciferase as a probe. From the bright-field images we obtained the local deflection angle to the vertical, A, local curvature, C, and the partial derivative of C with respect to time, [Formula: see text]. These were determined every 19.9 µm along the curvilinear length of the hypocotyl, at ~10 min intervals over a period of ~6 h after turning hypocotyls through 90° to the horizontal. Similarly from the luminescence images we measured the luminescence intensity of the convex and concave flanks of the hypocotyl as well as along the median of the hypocotyl, to determine differential expression of auxin-inducible IAA19. Comparison of these parameters as a function of time and curvilinear length shows that the gravitropic response is composed of three successive elements: the first and second curving responses and a decurving response (autostraightening). The maximum of the first curving response occurs when A is 76° along the entire length of the hypocotyl, suggesting that A is the sole determinant in this response; in contrast, the decurving response is a function of both A and C, as predicted by the newly-proposed graviproprioception model (Bastien et al., Proc Natl Acad Sci USA 110:755-760, 2013). Further, differential expression of IAA19, with higher expression in the convex flank, is observed at A = 44°, and follows the Sachs' sine law. This also suggests that IAA19 is not involved in the first curving response. In summary, the gravitropic response of Arabidopsis hypocotyls consists of multiple elements that are each determined by separate principles.

  19. Effects of Low Intensity Continuous Ultrasound (LICU on Mouse Pancreatic Tumor Explants

    Directory of Open Access Journals (Sweden)

    Despina Bazou

    2017-12-01

    Full Text Available This paper describes the effects of low intensity continuous ultrasound (LICU on the inflammatory response of mouse pancreatic tumor explants. While there are many reports focusing on the application of low-intensity pulsed ultrasound (LIPUS on cell cultures and tissues, the effects of continuous oscillations on biological tissues have never been investigated. Here we present an exploratory study of the effects induced by LICU on mouse pancreatic tumor explants. We show that LICU causes significant upregulation of IFN-γ, IL-1β, and TNF-α on tumor explants. No detectable effects were observed on tumor vasculature or collagen I deposition, while thermal and mechanical effects were not apparent. Tumor explants responded as a single unit to acoustic waves, with spatial pressure variations smaller than their size.

  20. In vitro direct regeneration in mint from different explants on half ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-09-15

    Sep 15, 2009 ... of medical practice in all countries of the world (Aftab and. Sial, 1999). ... mentha, have anti-feeding, insecticidal (Hori, 1999) anti- viral, antibacterial .... Internode explants were non responsive on all con- centrations of BAP.

  1. Distribution and utilization of nitrogenated compounds explanted by the soybean nodules by plants during seeds developing

    International Nuclear Information System (INIS)

    Alencar, Severino Matias de

    1997-01-01

    An experiment was carried out, using radioisotopes, for evaluation of the leaf, schuck and seeds areas and, examination of the pattern which is used by the nitrogenated compounds explanted by the soybean nodules

  2. Subcellular localization of H(+)-ATPase from pumpkin hypocotyls (Cucurbita maxima L.) by membrane fractionation.

    Science.gov (United States)

    Scherer, G F

    1984-03-01

    A new method of preparing sealed vesicles from membrane fractions of pumpkin hypocotyls in ethanolamine-containing buffers was used to investigate the subcellular localization of H(+)-ATPase measured as nigericin-stimulated ATPase. In a fluorescence-quench assay, the H(+) pump was directly demonstrated. The H(+) pump was substrate-specific for Mg·ATP and 0.1 mM diethylstilbestrol completely prevented the development of a Δ pH. The presence of unsupecific phosphatase hampered the detection of nigericin-stimulated ATPase. Unspecific phosphatases could be demonstrated by comparing the broad substrate specificity of the hydrolytic activities of the fractions with the clear preference for Mg·ATP as the substrate for the proton pump. Inhibitor studies showed that neither orthovanadate nor molybdate are absolutely specific for ATPase or acid phosphatase, respectively. Diethylstilbestrol seemed to be a specific inhibitor of ATPase activity in fractions containing nigericin-stimulated ATPase, but it stimulated acid phosphatase which tended to obscure its effect on ATPase activity. Nigericin-stimulated ATPase had its optimum at pH 6.0 and the nigericin effect was K(+)-dependent. The combination of valinomycin and carbonylcyanide m-chlorophenylhydrazone had a similar effect to nigericin, but singly these ionophores were much less stimulatory. After prolonged centrifugation on linear sucrose gradients, nigericin-stimulated ATPase correlated in dense fractions with plasma membrane markers but a part of it remained at the interphase. This lessdense part of the nigericin-stimulated ATPase could be derived from tonoplast vesicles because α-mannosidase, an enzyme of the vacuolar sap, remained in the upper part of the gradient. Nigericinstimulated ATPase did not correlate with the mitochondrial marker, cytochrome c oxidase, whereas azide inhibition of ATPase activity did.

  3. Efficient protocol for isolation and purification of different soyasaponins from soy hypocotyls.

    Science.gov (United States)

    Zhao, Dayun; Yan, Mingxia; Huang, Yuai; Sun, Xiangjun

    2012-12-01

    Soyasaponins are naturally occurring triterpenoid glycosides associated with many biological activities. The aim of the present study was to develop an effective method for isolation and purification of differently glycosylated, acetylated, and 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP)-conjugated soyasaponins from soy hypocotyls. Both gel filtration using Sephadex LH-20 chromatography (Amersham Pharmacia Biotech AB; elution phase: methanol, flow rate: 3.0 mL/min, sample loading: 60 mg) and high-speed countercurrent chromatography (stationary phase: n-butanol-acetic acid (5.0%, v/v), mobile phase: water flow rate: 3.0 mL/min, sample loading: 100 mg) could effectively fractionate isoflavones and soyasaponins from the crude extract with yield of soyasaponin complexes 20.5 mg and 22.3 mg, respectively. After fractionation, the soyasaponin complexes could be purified further using preparative HPLC to separate individuals. A total of nine soyasaponins, triacetyl soyasaponin Ab (yield 1.55%, HPLC purity >98%), Aa (2.68%, >99%), Ab (18.53%, >98%), Ae (0.85%, >98%), Ba (0.63%, >91%), Af (1.12%, >85%), Bb (3.45%, >98%) and Be (0.59%, >76.8%) were obtained. DDMP-conjugated groups, αg (2.06%, >85%), βg (7.59%, >85%), and γg (0.29%, >85%) that were very labile even in mild conditions, were also collected. The method described here can be used as an effective protocol to separate different soyasaponins occurring in the original sample. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Circadian clock gene LATE ELONGATED HYPOCOTYL directly regulates the timing of floral scent emission in Petunia.

    Science.gov (United States)

    Fenske, Myles P; Hewett Hazelton, Kristen D; Hempton, Andrew K; Shim, Jae Sung; Yamamoto, Breanne M; Riffell, Jeffrey A; Imaizumi, Takato

    2015-08-04

    Flowers present a complex display of signals to attract pollinators, including the emission of floral volatiles. Volatile emission is highly regulated, and many species restrict emissions to specific times of the day. This rhythmic emission of scent is regulated by the circadian clock; however, the mechanisms have remained unknown. In Petunia hybrida, volatile emissions are dominated by products of the floral volatile benzenoid/phenylpropanoid (FVBP) metabolic pathway. Here we demonstrate that the circadian clock gene P. hybrida LATE ELONGATED HYPOCOTYL (LHY; PhLHY) regulates the daily expression patterns of the FVBP pathway genes and floral volatile production. PhLHY expression peaks in the morning, antiphasic to the expression of P. hybrida GIGANTEA (PhGI), the master scent regulator ODORANT1 (ODO1), and many other evening-expressed FVBP genes. Overexpression phenotypes of PhLHY in Arabidopsis caused an arrhythmic clock phenotype, which resembles those of LHY overexpressors. In Petunia, constitutive expression of PhLHY depressed the expression levels of PhGI, ODO1, evening-expressed FVBP pathway genes, and FVBP emission in flowers. Additionally, in the Petunia lines in which PhLHY expression was reduced, the timing of peak expression of PhGI, ODO1, and the FVBP pathway genes advanced to the morning. Moreover, PhLHY protein binds to cis-regulatory elements called evening elements that exist in promoters of ODO1 and other FVBP genes. Thus, our results imply that PhLHY directly sets the timing of floral volatile emission by restricting the expression of ODO1 and other FVBP genes to the evening in Petunia.

  5. One-stage explant-implant procedure of exposed porous orbital implants

    DEFF Research Database (Denmark)

    Toft, Peter B; Rasmussen, Marie L Roed; Prause, Jan Ulrik

    2011-01-01

    Purpose:  To investigate the risks of implant exposure after a combined explant-implant procedure in patients with an exposed porous orbital implant. Methods:  Twenty-four consecutive patients who had a combined explant-implant procedure of an exposed hydroxyapatite (21) or porous polyethylene (3...... at the same procedure in sockets without profound signs of infection. The procedure carries a possible risk of poor motility....

  6. An early look at the Organ Procurement and Transplantation Network explant pathology form data.

    Science.gov (United States)

    Harper, Ann M; Edwards, Erick; Washburn, W Kenneth; Heimbach, Julie

    2016-06-01

    In April 2012, the Organ Procurement and Transplantation Network (OPTN) implemented an online explant pathology form for recipients of liver transplantation who received additional wait-list priority for their diagnosis of hepatocellular carcinoma (HCC). The purpose of the form was to standardize the data being reported to the OPTN, which had been required since 2002 but were submitted to the OPTN in a variety of formats via facsimile. From April 2012 to December 2014, over 4500 explant forms were submitted, allowing for detailed analysis of the characteristics of the explanted livers. Data from the explant pathology forms were used to assess agreement with pretransplant imaging. Explant data were also used to assess the risk of recurrence. Of those with T2 priority, 55.7% were found to be stage T2 on explant. Extrahepatic spread (odds ratio [OR] = 6.8; P based on the number and size of tumors on the explant form was T4 (OR = 2.4; P < 0.01) were the strongest predictors of recurrence. In conclusion, this analysis confirms earlier findings that showed an incomplete agreement between pretransplant imaging and posttransplant pathology in terms of HCC staging, though the number of patients with both no pretransplant treatment and no tumor in the explant was reduced from 20% to <1%. In addition, several factors were identified (eg, tumor burden, age, sex, region, ablative therapy, alpha-fetoprotein, Milan stage, vascular invasion, satellite lesions, etc.) that were predictive of HCC recurrence, allowing for more targeted surveillance of high-risk recipients. Continued evaluation of these data will help shape future guidelines or policy recommendations. Liver Transplantation 22 757-764 2016 AASLD. © 2016 American Association for the Study of Liver Diseases.

  7. Optic nerve compression as a late complication of a hydrogel explant with silicone encircling band.

    Science.gov (United States)

    Crama, Niels; Kluijtmans, Leo; Klevering, B Jeroen

    2018-06-01

    To present a complication of compressive optic neuropathy caused by a swollen hydrogel explant and posteriorly displaced silicone encircling band. A 72-year-old female patient presented with progressive visual loss and a tilted optic disc. Her medical history included a retinal detachment in 1993 that was treated with a hydrogel explant under a solid silicone encircling band. Visual acuity had decreased from 6/10 to 6/20 and perimetry showed a scotoma in the temporal superior quadrant. On Magnetic Resonance Imaging (MRI), compression of the optic nerve by a displaced silicone encircling band inferior nasally in combination with a swollen episcleral hydrogel explant was observed. Surgical removal of the hydrogel explant and silicone encircling band was uneventful and resulted in improvement of visual acuity and visual field loss. This is the first report on compressive optic neuropathy caused by swelling of a hydrogel explant resulting in a dislocated silicone encircling band. The loss of visual function resolved upon removal of the explant and encircling band.

  8. Isolation of carrot plant lines with altered carotene contents from gamma irradiated explants

    International Nuclear Information System (INIS)

    Pawlicki, N.; Sangwan, R.S.; Sangwan-Norreel, B.S.

    2001-01-01

    Dietary vitamin A is mainly obtained from carotenes of vegetables and fruits. Carrot (Daucus carota L.) is one of the major sources of carotene. Carrot cultivars have been obtained mainly through classical breeding, and genetic selection has permitted the creation of new varieties with high carotene contents. The fact that in several crops agronomically important mutants/variants have been generated by in vitro culture techniques prompted us to combine gamma irradiation and in vitro somatic embryogenesis to obtain regenerants with variations in carotene content in carrot. To test the effect of gamma rays on somatic embryogenesis and on the carotene level, aseptically germinated seedlings of 8 carrot varieties were exposed to 5; 10 and 500 Gy before culturing petiole segments on LN1 medium. Non-irradiated petioles produced calli with somatic embryos, while irradiated explants reacted differently according to radiation dose. After 4 weeks of culture on LN1 medium, petiole segments of different varieties irradiated with 5 and 10 Gy gave more callus with embryos than those with non-irradiated segments. However, after the subculture on LN medium, the development of embryos into plantlets was rare. It was also noted that after irradiation with 5 Gy, the petiole segments gave voluminous calli. Further, in variety 'Chantenay', the irradiated calli were deep orange while non-irradiated calli were green. However, embryo formation was not observed in these calli. This orange coloration suggests an appreciable synthesis of carotene in the calli. Gamma rays, probably produced cell lines with different colors and carotene content. Of the 8 cultivars tested, normal plantlets of 3 varieties were regenerated from somatic embryos irradiated with 10 Gy, and were transferred to greenhouse to develop roots. For each assay, the carotene analysis was carried out on 2 roots, and compared with plants produced from non-irradiated somatic embryos. Carotene level in the plants, derived from

  9. Brookhaven segment interconnect

    International Nuclear Information System (INIS)

    Morse, W.M.; Benenson, G.; Leipuner, L.B.

    1983-01-01

    We have performed a high energy physics experiment using a multisegment Brookhaven FASTBUS system. The system was composed of three crate segments and two cable segments. We discuss the segment interconnect module which permits communication between the various segments

  10. UV-B inhibition of hypocotyl growth in etiolated Arabidopsis thaliana seedlings is a consequence of cell cycle arrest initiated by photodimer accumulation

    Science.gov (United States)

    Biever, Jessica J.; Brinkman, Doug; Gardner, Gary

    2014-01-01

    Ultraviolet (UV) radiation is an important constituent of sunlight that determines plant morphology and growth. It induces photomorphogenic responses but also causes damage to DNA. Arabidopsis mutants of the endonucleases that function in nucleotide excision repair, xpf-3 and uvr1-1, showed hypersensitivity to UV-B (280–320nm) in terms of inhibition of hypocotyl growth. SOG1 is a transcription factor that functions in the DNA damage signalling response after γ-irradiation. xpf mutants that carry the sog1-1 mutation showed hypocotyl growth inhibition after UV-B irradiation similar to the wild type. A DNA replication inhibitor, hydroxyurea (HU), also inhibited hypocotyl growth in etiolated seedlings, but xpf-3 was not hypersensitive to HU. UV-B irradiation induced accumulation of the G2/M-specific cell cycle reporter construct CYCB1;1-GUS in wild-type Arabidopsis seedlings that was consistent with the expected accumulation of photodimers and coincided with the time course of hypocotyl growth inhibition after UV-B treatment. Etiolated mutants of UVR8, a recently described UV-B photoreceptor gene, irradiated with UV-B showed inhibition of hypocotyl growth that was not different from that of the wild type, but they lacked UV-B-specific expression of chalcone synthase (CHS), as expected from previous reports. CHS expression after UV-B irradiation was not different in xpf-3 compared with the wild type, nor was it altered after HU treatment. These results suggest that hypocotyl growth inhibition by UV-B light in etiolated Arabidopsis seedlings, a photomorphogenic response, is dictated by signals originating from UV-B absorption by DNA that lead to cell cycle arrest. This process occurs distinct from UVR8 and its signalling pathway responsible for CHS induction. PMID:24591052

  11. Full-thickness human skin explants for testing the toxicity of topically applied chemicals

    International Nuclear Information System (INIS)

    Nakamura, M.; Rikimaru, T.; Yano, T.; Moore, K.G.; Pula, P.J.; Schofield, B.H.; Dannenberg, A.M. Jr.

    1990-01-01

    This report describes a model organ-culture system for testing the toxicity of chemical substances that are topically applied to human skin. In this system, the viable keratinocytes in the full-thickness skin explants are protected by the same keratinized layer as skin remaining on the donor, and toxicity can be assessed microscopically and/or biochemically. The human skin specimens were discards from a variety of surgical procedures. They were cut into full-thickness 1.0-cm2 explants, and briefly exposed to the military vesicant sulfur mustard (SM), which was used as a model toxicant. The explants were then organ cultured in small Petri dishes for 24 h at 36 degrees C. In the 0.03-1.0% dosage range, a straight-line dose-response relationship occurred between the concentration of SM applied and the number of paranuclear vacuoles seen histologically in the epidermis. Within the same SM dosage range, there was also a proportional decrease in 14C-leucine incorporation by the explants. Thus, the number of paranuclear vacuoles reflected decreases in protein synthesis by the injured epidermal cells. The epidermis of full-thickness untreated (control) human skin explants usually remained viable for 7 d when stored at 4 degrees C in culture medium. During storage, a relatively small number of paranuclear vacuoles developed within the epidermis, but the explants were still quite satisfactory for testing SM toxicity. Incubation (for 4 or 24 h at 36 degrees C) of such control skin explants reduced (often by 50%) the small number of paranuclear vacuoles produced during 4-7 d of storage. This reduction was probably caused by autolysis of many of the vacuolated cells. Two types of paranuclear vacuoles could be identified by both light and electron microscopy: a storage type and a toxicant type. The storage type seemed to be caused by autolysis of cell components

  12. Full-thickness human skin explants for testing the toxicity of topically applied chemicals

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, M.; Rikimaru, T.; Yano, T.; Moore, K.G.; Pula, P.J.; Schofield, B.H.; Dannenberg, A.M. Jr. (Johns Hopkins Univ., Baltimore, MD (USA))

    1990-09-01

    This report describes a model organ-culture system for testing the toxicity of chemical substances that are topically applied to human skin. In this system, the viable keratinocytes in the full-thickness skin explants are protected by the same keratinized layer as skin remaining on the donor, and toxicity can be assessed microscopically and/or biochemically. The human skin specimens were discards from a variety of surgical procedures. They were cut into full-thickness 1.0-cm2 explants, and briefly exposed to the military vesicant sulfur mustard (SM), which was used as a model toxicant. The explants were then organ cultured in small Petri dishes for 24 h at 36 degrees C. In the 0.03-1.0% dosage range, a straight-line dose-response relationship occurred between the concentration of SM applied and the number of paranuclear vacuoles seen histologically in the epidermis. Within the same SM dosage range, there was also a proportional decrease in 14C-leucine incorporation by the explants. Thus, the number of paranuclear vacuoles reflected decreases in protein synthesis by the injured epidermal cells. The epidermis of full-thickness untreated (control) human skin explants usually remained viable for 7 d when stored at 4 degrees C in culture medium. During storage, a relatively small number of paranuclear vacuoles developed within the epidermis, but the explants were still quite satisfactory for testing SM toxicity. Incubation (for 4 or 24 h at 36{degrees}C) of such control skin explants reduced (often by 50%) the small number of paranuclear vacuoles produced during 4-7 d of storage. This reduction was probably caused by autolysis of many of the vacuolated cells. Two types of paranuclear vacuoles could be identified by both light and electron microscopy: a storage type and a toxicant type. The storage type seemed to be caused by autolysis of cell components.

  13. Encapsulation of nodal segments of lobelia chinensis

    Directory of Open Access Journals (Sweden)

    Weng Hing Thong

    2015-04-01

    Full Text Available Lobelia chinensis served as an important herb in traditional chinese medicine. It is rare in the field and infected by some pathogens. Therefore, encapsulation of axillary buds has been developed for in vitro propagation of L. chinensis. Nodal explants of L. chinensis were used as inclusion materials for encapsulation. Various combinations of calcium chloride and sodium alginate were tested. Encapsulation beads produced by mixing 50 mM calcium chloride and 3.5% sodium alginate supported the optimal in vitro conversion potential. The number of multiple shoots formed by encapsulated nodal segments was not significantly different from the average of shoots produced by non-encapsulated nodal segments. The encapsulated nodal segments regenerated in vitro on different medium. The optimal germination and regeneration medium was Murashige-Skoog medium. Plantlets regenerated from the encapsulated nodal segments were hardened, acclimatized and established well in the field, showing similar morphology with parent plants. This encapsulation technology would serve as an alternative in vitro regeneration system for L. chinensis.

  14. Manipulation of the hypocotyl sink activity by reciprocal grafting of two Raphanus sativus varieties: its effects on morphological and physiological traits of source leaves and whole-plant growth.

    Science.gov (United States)

    Sugiura, Daisuke; Betsuyaku, Eriko; Terashima, Ichiro

    2015-12-01

    To reveal whether hypocotyl sink activities are regulated by the aboveground parts, and whether physiology and morphology of source leaves are affected by the hypocotyl sink activities, we conducted grafting experiments using two Raphanus sativus varieties with different hypocotyl sink activities. Comet (C) and Leafy (L) varieties with high and low hypocotyl sink activities were reciprocally grafted and resultant plants were called by their scion and stock such as CC, LC, CL and LL. Growth, leaf mass per area (LMA), total non-structural carbohydrates (TNCs) and photosynthetic characteristics were compared among them. Comet hypocotyls in CC and LC grew well regardless of the scions, whereas Leafy hypocotyls in CL and LL did not. Relative growth rate was highest in LL and lowest in CC. Photosynthetic capacity was correlated with Rubisco (ribulose 1·5-bisphosphate carboxylase/oxygenase) content but unaffected by TNC. High C/N ratio and accumulation of TNC led to high LMA and structural LMA. These results showed that the hypocotyl sink activity was autonomously regulated by hypocotyl and that the down-regulation of photosynthesis was not induced by TNC. We conclude that the change in the sink activity alters whole-plant growth through the changes in both biomass allocation and leaf morphological characteristics in R. sativus. © 2015 John Wiley & Sons Ltd.

  15. Genome-wide binding site analysis of FAR-RED ELONGATED HYPOCOTYL3 reveals its novel function in Arabidopsis development.

    Science.gov (United States)

    Ouyang, Xinhao; Li, Jigang; Li, Gang; Li, Bosheng; Chen, Beibei; Shen, Huaishun; Huang, Xi; Mo, Xiaorong; Wan, Xiangyuan; Lin, Rongcheng; Li, Shigui; Wang, Haiyang; Deng, Xing Wang

    2011-07-01

    FAR-RED ELONGATED HYPOCOTYL3 (FHY3) and its homolog FAR-RED IMPAIRED RESPONSE1 (FAR1), two transposase-derived transcription factors, are key components in phytochrome A signaling and the circadian clock. Here, we use chromatin immunoprecipitation-based sequencing (ChIP-seq) to identify 1559 and 1009 FHY3 direct target genes in darkness (D) and far-red (FR) light conditions, respectively, in the Arabidopsis thaliana genome. FHY3 preferentially binds to promoters through the FHY3/FAR1 binding motif (CACGCGC). Interestingly, FHY3 also binds to two motifs in the 178-bp Arabidopsis centromeric repeats. Comparison between the ChIP-seq and microarray data indicates that FHY3 quickly regulates the expression of 197 and 86 genes in D and FR, respectively. FHY3 also coregulates a number of common target genes with PHYTOCHROME INTERACTING FACTOR 3-LIKE5 and ELONGATED HYPOCOTYL5. Moreover, we uncover a role for FHY3 in controlling chloroplast development by directly activating the expression of ACCUMULATION AND REPLICATION OF CHLOROPLASTS5, whose product is a structural component of the latter stages of chloroplast division in Arabidopsis. Taken together, our data suggest that FHY3 regulates multiple facets of plant development, thus providing insights into its functions beyond light and circadian pathways.

  16. Plant regeneration from cotyledonary explants of Eucalyptus camaldulensis Regeneração de plantas de Eucalyptus camaldulensis a partir das explantes cotiledonares

    Directory of Open Access Journals (Sweden)

    Roberson Dibax

    2005-08-01

    Full Text Available Breeding methods based on genetic transformation techniques need to be implemented for Eucalyptus camaldulensis to shorten the long breeding cycles and avoid manipulation of adult trees; that requires the development of plant regeneration protocols enabling development of plants from transformed tissues. The present work aimed to optimise the regeneration process already established for the species. Cotyledonary leaves of E. camaldulensis were cultured in MS medium supplemented with naphthaleneacetic acid (NAA and 6-benzylaminopurine (BAP combinations. The most efficient treatment for bud indirect regeneration (2.7 µmol L-1 NAA and 4.44 µmol L-1 BAP was used for further experiments. When explants were kept in the dark during the first 30 days, the percentage of explants forming calluses increased and explant necrosis was reduced in comparison with light-cultured explants. Mineral medium modifications were compared and half-strength MS mineral medium turned out to be as efficient as full-strength medium, producing 54% and 47% of explants with buds, respectively. For shoot elongation, MS medium with half-strength nitrate and ammonium salts, and 0.2% activated charcoal yielded rooted shoots 1 to 8 cm high after one month. The procedure is an efficient protocol for E. camadulensis plant regeneration, reducing the stages necessary for the obtention of complete plants.A implementação, para espécies florestais, de técnicas de melhoramento baseadas em métodos de transformação genética, permitirá reduzir os longos ciclos de melhoramento e evitar a manipulação de árvores adultas. Isto implica dispor de um protocolo de regeneração que permita o desenvolvimento de plantas a partir de tecidos transformados. Este trabalho teve como objetivo otimizar este protocolo de regeneração para Eucalyptus camaldulensis. Folhas cotiledonares foram cultivadas em meio de cultura MS suplementado com combinações de ácido naftalenoacético (ANA e 6

  17. 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.

  18. Organogênese de explante foliar de clones de Eucalyptus grandis x E. urophylla Organogenesis of the leaf explant of Eucalyptus grandis x E. urophylla clones

    Directory of Open Access Journals (Sweden)

    Elisa Cristina Soares de Carvalho Alves

    2004-05-01

    Full Text Available O objetivo deste trabalho foi avaliar os efeitos dos reguladores de crescimento TDZ [1-fenil-3-(1,2,3-tia-diazol-5-iluréia], BAP (6-benzilaminopurina e ANA (ácido naftalenoacético no desempenho da propagação in vitro por organogênese de explante foliar de três clones híbridos de Eucalyptus grandis x Eucalyptus urophylla. Houve resposta diferenciada dos clones quanto a intensidade, textura e coloração dos calos, em razão dos tratamentos com os reguladores de crescimento. Os melhores resultados de calejamento dos três genótipos foram observados nos tratamentos com a combinação dos reguladores de crescimento TDZ (0,5 mg L-1 e ANA (0,1 mg L-1, obtendo-se 100% de calejamento no explante foliar. Os piores resultados de calejamento foram observados nos tratamentos com a combinação dos reguladores de crescimento BAP (0,1 mg L-1 e ANA (0,1 mg L-1. Em relação à regeneração, a melhor resposta foi obtida com 1,0 mg L-1 BAP em que 8% dos calos formados a partir de explantes foliares regeneraram gemas, com número médio destas formadas por calo igual a 4,2.The aim of this work was to evaluate the effects of growth regulators TDZ [1-phenil-3-(1,2,3-thiadiazol-5-yl urea], BAP (6-benzilaminopurine e NAA (Naphthalene acetic acid on the in vitro propagation by organogenesis from foliar explants of Eucalyptus grandis x E. urophylla. Depending on the clone used, there were singular responses to growth regulators treatment regarding callusing intensity, texture and color. The best results of the three genotypes used were observed with the TDZ (0.5 mg L-1 and NAA (0.1 mg L-1 treatment, where 100% of the foliar explants presented callus. The worst results were observed with the BAP (0.1 mg L-1 and NAA (0.1 mg L-1 treatment. Subsequently, considering the regeneration process, the best response was achieved with 1.0 mg L-1 BAP, in which 8% of the calli regenerated buds, with an average of 4.2 buds per explant.

  19. Resolving browning during the establishment of explant cultures in Vicia faba L. for genetic transformation

    Directory of Open Access Journals (Sweden)

    Helena Klenotičová

    2013-01-01

    Full Text Available Optimisation of in vitro regeneration systems of two explant types for low-tannine cultivars of faba bean based on culturing of shoot apices and cotyledonary nodes were provided by usage of various antioxidants - ascorbic acid, citric acid, glutathione and activated charcoal. In subsequent testing, the combined effects of antioxidants with transformation co-cultivation compounds acetosyringone and L-cysteine was studied. The application of antioxidants lead to decreased callogenesis, citric acids treatments (50 mg.l−1 dramatically decreased necrotic response of explants. However, citric acid, used together with ascorbic acid completely inhibited shoot growth in shoot apex cultures. Glutathion evoked hyperhydricity of explants. Activated charcoal induced rooting on media which are commonly used for shoot proliferation. Combination of acetosyringone with antioxidants influenced shoot proliferation, except of variant with ascorbic acid. Citric acid was the best and universal antioxidant in faba bean in vitro cultures and its use is recommended for faba bean genetic transformation experiments.

  20. Explant culture of human peripheral lung. I. Metabolism of benzo[alpha]pyrene

    DEFF Research Database (Denmark)

    Stoner, G.D.; Harris, C.C.; Autrup, Herman

    1978-01-01

    the predominant alveolar epithelial cell type. Lamellar inclusion bodies were released from the type 2 cells and accumulated in the alveolar spaces. The metabolism of benzo[alpha]pyrene (BP) in human lung explants cultured for up to 7 days was investigated. Human lung explants had measurable aryl hydrocarbon......Human lung explants have been maintained in vitro for a period of 25 days. Autoradiographic studies indicated that the broncholar epithelial cells, type 2 alveolar epithelial cells, and stromal fibroblasts incorporated 3H-thymidine during the culture. After 7 to 10 days, type 2 cells were...... hydroxylase activity and could metabolize BP into forms that were bound to cellular DNA and protein. Peripheral lung had significantly lower aryl hydrocarbon hydroxylase activity than cultured bronchus but both tissues had similar binding levels of BP to DNA. Radioautographic studies indicated that all cell...

  1. Somatic embryogenesis from leaf explants of Australian fan flower, Scaevola aemula R. Br.

    Science.gov (United States)

    Wang, Y-H; Bhalla, P L

    2004-01-01

    Somatic embryogenesis from leaf explants of Scaevola aemula R. Br. was achieved. Somatic embryos were induced from explants cultured on MS medium supplemented with 0.2 mg/ 2,4-dichlorophenoxyacetic acid and 0.2-0.5 mg/l 6-benzylaminopurine (BAP). Various developmental stages of somatic embryos were found on this medium-from globular embryos to germinated embryos. The transfer of globular embryos to MS medium containing 0.5 mg/l BAP resulted in a high frequency of shoot regeneration. Leaf explants cultured on MS medium containing different combinations of BAP and alpha-naphthaleneacetic acid formed adventitious shoots and roots. Histological examination confirmed the process of somatic embryogenesis. Induction of somatic embryogenesis in Scaevola provides a system for studying embryogenesis in Australian native plants and will facilitate the improvement of these plants using genetic transformation techniques.

  2. Sprifermin (rhFGF18) modulates extracellular matrix turnover in cartilage explants ex vivo

    DEFF Research Database (Denmark)

    Reker, Ditte; Kjelgaard-Petersen, Cecilie Freja; Siebuhr, Anne Sofie

    2017-01-01

    (ECM) production. To gain further insight into the process of sprifermin in the cartilage tissue, this study aimed at investigating the ECM turnover of articular cartilage explants in a longitudinal manner. Methods: Bovine full-depth articular cartilage explants were stimulated with sprifermin...... by immuno-histochemical detection of proliferating cell nuclear antigen. ECM turnover was quantified by biomarker ELISAs; ProC2 reflecting type II collagen formation, CS846 reflecting aggrecan formation, active MMP9, C2M and AGNx2 reflecting matrix metalloproteinase activity, and AGNx1 reflecting......, active MMP9 was slightly decreased, and AGNx1 was slightly increased. Over the course of treatment, the temporal order of ECM turnover responses was AGNx1, then ProC2, followed by CS846 and MMP9. Pro-inflammatory activation of the explants diminished the ECM turnover responses otherwise observed under...

  3. BACTERIAL CONTAMINATION CONTROL IN BANANA EXPLANTS (Musa AAA cv. CAIPIRA) CONTROLE DE BACTÉRIAS CONTAMINANTES EM EXPLANTES DE BANANEIRA (Musa AAA cv. CAIPIRA)

    OpenAIRE

    Juliana Domingues Lima; Wilson da Silva Moraes

    2007-01-01

    Esse trabalho teve por objetivo testar métodos de controle de contaminação bacteriana no processo de multiplicação in vitro de bananeira (Musa AAA cv. Caipira), utilizando-se hipoclorito de sódio (NaOCl), antibiótico rifampicina e suas combinações. Não houve oxidação excessiva dos explantes após a imersão em NaOCl ou rifampicina. O melhor tratamento para explantes recém isolados foi imersão em NaOCl a 1% (v/v), dura...

  4. Patient-specific three-dimensional explant spheroids derived from human nasal airway epithelium

    DEFF Research Database (Denmark)

    Marthin, June Kehlet; Stevens, Elizabeth Munkebjerg; Larsen, Lars Allan

    2017-01-01

    BACKGROUND: Three-dimensional explant spheroid formation is an ex vivo technique previously used in studies of airway epithelial ion and water transport. Explanted cells and sheets of nasal epithelium form fully differentiated spheroids enclosing a partly fluid-filled lumen with the ciliated apical...... surface facing the outside and accessible for analysis of ciliary function. METHODS: We performed a two-group comparison study of ciliary beat pattern and ciliary beat frequency in spheroids derived from nasal airway epithelium in patients with primary ciliary dyskinesia (PCD) and in healthy controls...... in the investigation of pathophysiological aspects and drug effects in human nasal airway epithelium....

  5. Transport of Indole-3-Butyric Acid and Indole-3-Acetic Acid in Arabidopsis Hypocotyls Using Stable Isotope Labeling1[C][W][OA

    Science.gov (United States)

    Liu, Xing; Barkawi, Lana; Gardner, Gary; Cohen, Jerry D.

    2012-01-01

    The polar transport of the natural auxins indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA) has been described in Arabidopsis (Arabidopsis thaliana) hypocotyls using radioactive tracers. Because radioactive assays alone cannot distinguish IBA from its metabolites, the detected transport from applied [3H]IBA may have resulted from the transport of IBA metabolites, including IAA. To test this hypothesis, we used a mass spectrometry-based method to quantify the transport of IBA in Arabidopsis hypocotyls by following the movement of [13C1]IBA and the [13C1]IAA derived from [13C1]IBA. We also assayed [13C6]IAA transport in a parallel control experiment. We found that the amount of transported [13C1]IBA was dramatically lower than [13C6]IAA, and the IBA transport was not reduced by the auxin transport inhibitor N-1-naphthylphthalamic acid. Significant amounts of the applied [13C1]IBA were converted to [13C1]IAA during transport, but [13C1]IBA transport was independent of IBA-to-IAA conversion. We also found that most of the [13C1]IBA was converted to ester-linked [13C1]IBA at the apical end of hypocotyls, and ester-linked [13C1]IBA was also found in the basal end at a level higher than free [13C1]IBA. In contrast, most of the [13C6]IAA was converted to amide-linked [13C6]IAA at the apical end of hypocotyls, but very little conjugated [13C6]IAA was found in the basal end. Our results demonstrate that the polar transport of IBA is much lower than IAA in Arabidopsis hypocotyls, and the transport mechanism is distinct from IAA transport. These experiments also establish a method for quantifying the movement of small molecules in plants using stable isotope labeling. PMID:22323783

  6. Transient gibberellin application promotes Arabidopsis thaliana hypocotyl cell elongation without maintaining transverse orientation of microtubules on the outer tangential wall of epidermal cells

    KAUST Repository

    Sauret-Güeto, Susanna

    2011-11-25

    The phytohormone gibberellin (GA) promotes plant growth by stimulating cellular expansion. Whilst it is known that GA acts by opposing the growth-repressing effects of DELLA proteins, it is not known how these events promote cellular expansion. Here we present a time-lapse analysis of the effects of a single pulse of GA on the growth of Arabidopsis hypocotyls. Our analyses permit kinetic resolution of the transient growth effects of GA on expanding cells. We show that pulsed application of GA to the relatively slowly growing cells of the unexpanded light-grown Arabidopsis hypocotyl results in a transient burst of anisotropic cellular growth. This burst, and the subsequent restoration of initial cellular elongation rates, occurred respectively following the degradation and subsequent reappearance of a GFP-tagged DELLA (GFP-RGA). In addition, we used a GFP-tagged α-tubulin 6 (GFP-TUA6) to visualise the behaviour of microtubules (MTs) on the outer tangential wall (OTW) of epidermal cells. In contrast to some current hypotheses concerning the effect of GA on MTs, we show that the GA-induced boost of hypocotyl cell elongation rate is not dependent upon the maintenance of transverse orientation of the OTW MTs. This confirms that transverse alignment of outer face MTs is not necessary to maintain rapid elongation rates of light-grown hypocotyls. Together with future studies on MT dynamics in other faces of epidermal cells and in cells deeper within the hypocotyl, our observations advance understanding of the mechanisms by which GA promotes plant cell and organ growth. © 2011 Blackwell Publishing Ltd.

  7. Cryopreservation of in vitro grown nodal segments of Rauvolfia serpentina by PVS2 vitrification.

    Science.gov (United States)

    Ray, Avik; Bhattacharya, Sabita

    2008-01-01

    This paper describes the cryopreservation by PVS2 vitrification of Rauvolfia serpentina (L.) Benth ex kurz, an important tropical medicinal plant. The effects of type and size of explants, sucrose preculture (duration and concentration) and vitrification treatment were tested. Preliminary experiments with PVS1, 2 and 3 produced shoot growth only for PVS2. When optimizing the PVS2 vitrification of nodal segments, those of 0.31 - 0.39 cm in size were better than other nodal sizes and or apices. Sucrose preculture had a positive role in survival and subsequent regrowth of the cryopreserved explants. Seven days on 0.5 M sucrose solution significantly improved the viability of nodal segments. PVS2 incubation for 45 minutes combined with a 7-day preculture gave the optimum result of 66 percent. Plantlets derived after cryopreservation resumed growth and regenerated normally.

  8. Low-intensity pulsed ultrasound (LIPUS) and pulsed electromagnetic field (PEMF) treatments affect degeneration of cultured articular cartilage explants

    NARCIS (Netherlands)

    Tan, Lijun; Ren, Yijin; van Kooten, Theo G.; Grijpma, Dirk W.; Kuijer, Roelof

    PURPOSE: Articular cartilage has some capacity for self-repair. Clinically used low-intensity pulsed ultrasound (LIPUS) and pulsed electromagnetic field (PEMF) treatments were compared in their potency to prevent degeneration using an explant model of porcine cartilage. METHODS: Explants of porcine

  9. Low-intensity pulsed ultrasound (LIPUS) and pulsed electromagnetic field (PEMF) treatments affect degeneration of cultured articular cartilage explants

    NARCIS (Netherlands)

    Tan, Lijun; Tan, Lijun; Ren, Yijin; van Kooten, Theo G.; Grijpma, Dirk W.; Kuijer, Roel

    2015-01-01

    Purpose: Articular cartilage has some capacity for self-repair. Clinically used low-intensity pulsed ultrasound (LIPUS) and pulsed electromagnetic field (PEMF) treatments were compared in their potency to prevent degeneration using an explant model of porcine cartilage. Methods: Explants of porcine

  10. An in vitro model for detecting skin irritants: methyl green-pyronine staining of human skin explant cultures

    NARCIS (Netherlands)

    Jacobs, J. J. L.; Lehé, C.; Cammans, K. D. A.; Das, P. K.; Elliott, G. R.

    2002-01-01

    We evaluated the potential of human organotypic skin explant cultures (hOSECs) for screening skin irritants. Test chemicals were applied to the epidermis of the skin explants which were incubated for 4, 24 or 48 h in tissue culture medium. A decrease in epidermal RNA staining, visualised in frozen

  11. Induction of bulb organogenesis in in vitro cultures of tarda tulip (Tulipa tarda Stapf.) from seed-derived explants.

    Science.gov (United States)

    Maślanka, Małgorzata; Bach, Anna

    2014-01-01

    A protocol for obtaining bulbs via in vitro organogenesis was developed for tarda tulip ( Tulipa tarda Stapf). Scale explants were obtained from bulbs formed at the base of seedlings or from adventitious bulbs that developed from callus tissue forming on stolons or on germinating seeds. Some explants were subjected to chilling at 5°C for 12 wk. The culture media contained 3 or 6% sucrose and was supplemented with either no growth regulators, either 0.5 μM 6-benzyl-aminopurine (BAP) or 18.9 or 94.6 μM abscisic acid (ABA). Cultures were maintained in the dark at 20°C. Callus tissue developed mainly on media without growth regulators or with BAP. Callus was formed from up to 96% of explants derived from non-chilled adventitious bulbs that were treated with 3% sucrose and 0.5 μM BAP. Less callus was formed from chilled explants compared with non-chilled explants. Newly formed adventitious bulbs appeared on the explants via direct and indirect organogenesis. The media with BAP promoted the formation of adventitious bulbs at a rate of 56-92% from non-chilled explants, whereas a maximum rate of 36% was observed from chilled explants. ABA inhibited the induction of adventitious bulbs and callus. The adventitious bulbs obtained in these experiments contained a meristem, which was evidence that they had developed properly.

  12. Cytoplasmic Calcium Increases in Response to Changes in the Gravity Vector in Hypocotyls and Petioles of Arabidopsis Seedlings1

    Science.gov (United States)

    Toyota, Masatsugu; Furuichi, Takuya; Tatsumi, Hitoshi; Sokabe, Masahiro

    2008-01-01

    Plants respond to a large variety of environmental signals, including changes in the gravity vector (gravistimulation). In Arabidopsis (Arabidopsis thaliana) seedlings, gravistimulation is known to increase the cytoplasmic free calcium concentration ([Ca2+]c). However, organs responsible for the [Ca2+]c increase and the underlying cellular/molecular mechanisms remain to be solved. In this study, using Arabidopsis seedlings expressing apoaequorin, a Ca2+-sensitive luminescent protein in combination with an ultrasensitive photon counting camera, we clarified the organs where [Ca2+]c increases in response to gravistimulation and characterized the physiological and pharmacological properties of the [Ca2+]c increase. When the seedlings were gravistimulated by turning 180°, they showed a transient biphasic [Ca2+]c increase in their hypocotyls and petioles. The second peak of the [Ca2+]c increase depended on the angle but not the speed of rotation, whereas the initial peak showed diametrically opposite characters. This suggests that the second [Ca2+]c increase is specific for changes in the gravity vector. The potential mechanosensitive Ca2+-permeable channel (MSCC) inhibitors Gd3+ and La3+, the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA), and the endomembrane Ca2+-permeable channel inhibitor ruthenium red suppressed the second [Ca2+]c increase, suggesting that it arises from Ca2+ influx via putative MSCCs in the plasma membrane and Ca2+ release from intracellular Ca2+ stores. Moreover, the second [Ca2+]c increase was attenuated by actin-disrupting drugs cytochalasin B and latrunculin B but not by microtubule-disrupting drugs oryzalin and nocodazole, implying that actin filaments are partially involved in the hypothetical activation of Ca2+-permeable channels. These results suggest that the second [Ca2+]c increase via MSCCs is a gravity response in the hypocotyl and petiole of Arabidopsis seedlings. PMID:18055589

  13. Studies on the effect of genotype and explant type on callogenesis and organogenesis in indica rice

    International Nuclear Information System (INIS)

    Noor, A.; Rashid, H.; Khan, M.H.; Chaudhry, Z.

    2011-01-01

    The purpose of the research study is to select the best rice explant and variety for Agrobacterium mediated genetic transformation studies. In vitro callus induction and regeneration frequency of two explants i.e. immature embryo and media supplemented mature grains of 9 rice varieties (Oryza sativa L.) was investigated by culturing explants source on N/sub 6/ with 2, 4-D at 2 mg/l for callus induction. Results indicated that mature seed explant produced significantly high number of calli as compared to immature embryo. Two types of calli were distinguished, designated as type-I calli and type-II calli. Regarding the quality of callus, type-I calli (produced from scutellum of mature grains) which were embryogenic produced higher plant regeneration frequency than type-II calli which were non-embryogenic. Mature-seed scutella calli and immature embryo-derived calli (after three weeks of culture) of rice varieties were transferred on regeneration medium i.e., MS salts and vitamins, 3% sucrose, 3% sorbitol, 2 g/l casine hydrolysate, NAA 1.0 mg/l, kin 2.5 mg /l and BAP at 0.5mg/l. The highest regeneration capacity was observed in DR-83 from mature seed derived calli followed by Basmati 385. After hardening the plantlets were transferred to soil. (author)

  14. Fumonisin B₁ (FB₁) Induces Lamellar Separation and Alters Sphingolipid Metabolism of In Vitro Cultured Hoof Explants.

    Science.gov (United States)

    Reisinger, Nicole; Dohnal, Ilse; Nagl, Veronika; Schaumberger, Simone; Schatzmayr, Gerd; Mayer, Elisabeth

    2016-03-24

    One of the most important hoof diseases is laminitis. Yet, the pathology of laminitis is not fully understood. Different bacterial toxins, e.g. endotoxins or exotoxins, seem to play an important role. Additionally, ingestion of mycotoxins, toxic secondary metabolites of fungi, might contribute to the onset of laminitis. In this respect, fumonsins are of special interest since horses are regarded as species most susceptible to this group of mycotoxins. The aim of our study was to investigate the influence of fumonisin B₁ (FB₁) on primary isolated epidermal and dermal hoof cells, as well as on the lamellar tissue integrity and sphingolipid metabolism of hoof explants in vitro. There was no effect of FB₁ at any concentration on dermal or epidermal cells. However, FB₁ significantly reduced the separation force of explants after 24 h of incubation. The Sa/So ratio was significantly increased in supernatants of explants incubated with FB₁ (2.5-10 µg/mL) after 24 h. Observed effects on Sa/So ratio were linked to significantly increased sphinganine concentrations. Our study showed that FB₁ impairs the sphingolipid metabolism of explants and reduces lamellar integrity at non-cytotoxic concentrations. FB₁ might, therefore, affect hoof health. Further in vitro and in vivo studies are necessary to elucidate the effects of FB₁ on the equine hoof in more detail.

  15. Somatic Embryogenesis in Peach-Palm (Bactris gasipaes) Using Different Explant Sources.

    Science.gov (United States)

    Steinmacher, Douglas A; Heringer, Angelo Schuabb; Jiménez, Víctor M; Quoirin, Marguerite G G; Guerra, Miguel P

    2016-01-01

    Peach palm (Bactris gasipaes Kunth) is a member of the family Arecaceae and is a multipurpose but underutilized species. Nowadays, fruit production for subsistence and local markets, and heart-of-palm production for local, national, and international markets are the most important uses of this plant. Conventional breeding programs in peach palm are long-term efforts due to the prolonged generation time, large plant size, difficulties with controlled pollination and other factors. Although it is a caespitose palm, its propagation is currently based on seeds, as off-shoots are difficult to root. Hence, tissue culture techniques are considered to be the most likely strategy for efficient clonal plantlet regeneration of this species. Among various techniques, somatic embryogenesis offers the advantages of potential automated large-scale production and putative genetic stability of the regenerated plantlets. The induction of somatic embryogenesis in peach palm can be achieved by using different explant sources including zygotic embryos, immature inflorescences and thin cell layers from the young leaves and shoot meristems. The choice of a particular explant depends on whether clonal propagation is desired or not, as well as on the plant conditions and availability of explants. Protocols to induce and express somatic embryogenesis from different peach palm explants, up to acclimatization of plantlets, are described in this chapter.

  16. Ex-plant consequence assessment for NUREG-1150: models, typical results, uncertainties

    International Nuclear Information System (INIS)

    Sprung, J.L.

    1988-01-01

    The assessment of ex-plant consequences for NUREG-1150 source terms was performed using the MELCOR Accident Consequence Code System (MACCS). This paper briefly discusses the following elements of MACCS consequence calculations: input data, phenomena modeled, computational framework, typical results, controlling phenomena, and uncertainties. Wherever possible, NUREG-1150 results will be used to illustrate the discussion. 28 references

  17. Production of immunoglobulins in gingival tissue explant cultures from juvenile periodontitis patients

    International Nuclear Information System (INIS)

    Hall, E.R.; Falkler, W.A. Jr.; Suzuki, J.B.

    1990-01-01

    B lymphocytes and plasma cells are histologically observed in granulomatous periodontal tissues of juvenile periodontitis (JP) patients. Local immune processes may participate in protective or immunopathologic roles in the pathogenesis of this disease. An in vitro explant culture system was utilized to demonstrate the production of immunoglobulins by diseased JP tissues. Immunodiffusion studies using goat anti-human gamma, alpha, or mu chain serum revealed IgG to be the major immunoglobulin present in 92% of the day 1 supernatant fluids (SF) of the 47 JP gingival tissue explant cultures. IgA was present in 15% of the SF; however, no IgM was detected. Staph Protein A isolated 14C-labeled IgG from the SF, when allowed to react with goat anti-human gamma chain serum, formed lines of precipitation. Positive autoradiographs confirmed the biosynthesis of IgG by the explant cultures. The in vitro gingival tissue explant culture system described provides a useful model for the study of localized immunoglobulins produced by diseased tissues of JP patients

  18. Adventitious bud formation from bulb-scale explants of Lilium speciosum Thunb. in vitro

    NARCIS (Netherlands)

    Aartrijk, van J.

    1984-01-01

    In this thesis the interactive effects are described of tissue, medium, and other environmental factors on the process of adventitious bud formation in vitro from bulb-scale explants of Lilium speciosum Thunb. Besides, results are presented of experiments

  19. Growth Response of Explants of Irvingia Gabonensis (O'rorke, Baill ...

    African Journals Online (AJOL)

    Growth response of explants of Irvingia gabonensis to in vitro treatment was investigated using full, half and one quarter strength mineral components based on Murashige and Skoog medium. Plant growth regulator (kinetin-Kin) with concentration levels of 0, 1, 2, 3, 4 and 5mg/l were used for shoots initiation, while axillary ...

  20. Transgenic carnation plants obtained by Agrobacterium tumefaciens mediated transformation of petal explants

    NARCIS (Netherlands)

    Altvorst, van A.C.; Koehorst, H.; Jong, de J.; Dons, M.M.

    1996-01-01

    Transgenic carnation plants were obtained after infection of petal explants with the supervirulent Agrobacterium tumefaciens strain AGLO. Southern blot techniques confirmed the transgenic nature of four transformed plants. The expression of the gus gene was verified in these plants by histochemical

  1. Effects of plant growth regulators on in vitro cultured nodal explants ...

    African Journals Online (AJOL)

    SAM

    2014-07-09

    Jul 9, 2014 ... sterilized potting mix of forest soil, well-decomposed coffee husk, and red sand at the respective ratio of 1:1:2 by volume (JARC experience) and the stock plants were established in the main- tenance greenhouse of the biotechnology division at an average temperature of 25 ± 2°C. Explant sterilization and ...

  2. A procedure for culturing rat neocortex explants in a serum-free nutrient medium

    NARCIS (Netherlands)

    Romijn, H. J.; de Jong, B. M.; Ruijter, J. M.

    1988-01-01

    A procedure is described for long-term culturing of rat neocortex explants in a serum-free growth medium. Slices spanning the entire cortical depth from pial to ventricular side are prepared from 6-day-old rat pups. After preincubation in Hanks' balanced salt solution with extra glucose, the

  3. Hypoxia preferentially destroys GABAergic neurons in developing rat neocortex explants in culture

    NARCIS (Netherlands)

    Romijn, H. J.; Ruijter, J. M.; Wolters, P. S.

    1988-01-01

    The hypothesis that hypoxic ischemia before or during the human birth process preferentially destroys GABAergic nerve cells, particularly in the neocortex, was tested in a tissue culture model system. To that end, rat neocortex explants dissected from 6-day-old rat pups and cultured to a

  4. Immunocytochemical indications for neuronal co-localization of GABA and aspartate in cultured neocortex explants

    NARCIS (Netherlands)

    de Jong, B. M.; Ruijter, J. M.; Buijs, R. M.

    1989-01-01

    The application of postembedding immunocytochemistry on serial semithin plastic sections, revealed the presence of gamma-aminobutyric acid (GABA)-positive and aspartate-positive neurons in cultured neocortex explants. GABA-positive neurons were found in all layers of the cultured cortex, whereas

  5. Production of immunoglobulins in gingival tissue explant cultures from juvenile periodontitis patients

    Energy Technology Data Exchange (ETDEWEB)

    Hall, E.R.; Falkler, W.A. Jr.; Suzuki, J.B. (Univ. of Maryland Dental School, Baltimore (USA))

    1990-10-01

    B lymphocytes and plasma cells are histologically observed in granulomatous periodontal tissues of juvenile periodontitis (JP) patients. Local immune processes may participate in protective or immunopathologic roles in the pathogenesis of this disease. An in vitro explant culture system was utilized to demonstrate the production of immunoglobulins by diseased JP tissues. Immunodiffusion studies using goat anti-human gamma, alpha, or mu chain serum revealed IgG to be the major immunoglobulin present in 92% of the day 1 supernatant fluids (SF) of the 47 JP gingival tissue explant cultures. IgA was present in 15% of the SF; however, no IgM was detected. Staph Protein A isolated 14C-labeled IgG from the SF, when allowed to react with goat anti-human gamma chain serum, formed lines of precipitation. Positive autoradiographs confirmed the biosynthesis of IgG by the explant cultures. The in vitro gingival tissue explant culture system described provides a useful model for the study of localized immunoglobulins produced by diseased tissues of JP patients.

  6. Effect of glutaraldehyde fixation on the frictional response of immature bovine articular cartilage explants.

    Science.gov (United States)

    Oungoulian, Sevan R; Hehir, Kristin E; Zhu, Kaicen; Willis, Callen E; Marinescu, Anca G; Merali, Natasha; Ahmad, Christopher S; Hung, Clark T; Ateshian, Gerard A

    2014-02-07

    This study examined functional properties and biocompatibility of glutaraldehyde-fixed bovine articular cartilage over several weeks of incubation at body temperature to investigate its potential use as a resurfacing material in joint arthroplasty. In the first experiment, treated cartilage disks were fixed using 0.02, 0.20 and 0.60% glutaraldehyde for 24h then incubated, along with an untreated control group, in saline for up to 28d at 37°C. Both the equilibrium compressive and tensile moduli increased nearly twofold in treated samples compared to day 0 control, and remained at that level from day 1 to 28; the equilibrium friction coefficient against glass rose nearly twofold immediately after fixation (day 1) but returned to control values after day 7. Live explants co-cultured with fixed explants showed no quantitative difference in cell viability over 28d. In general, no significant differences were observed between 0.20 and 0.60% groups, so 0.20% was deemed sufficient for complete fixation. In the second experiment, cartilage-on-cartilage frictional measurements were performed under a migrating contact configuration. In the treated group, one explant was fixed using 0.20% glutaraldehyde while the apposing explant was left untreated; in the control group both explants were left untreated. From day 1 to 28, the treated group exhibited either no significant difference or slightly lower friction coefficient than the untreated group. These results suggest that a properly titrated glutaraldehyde treatment can reproduce the desired functional properties of native articular cartilage and maintain these properties for at least 28d at body temperature. © 2013 Published by Elsevier Ltd.

  7. Direct and Indirect Somatic Embryogenesis from Petiole and Leaf Explants of Purple Fan Flower (Scaevola aemula R. Br. cv. 'Purple Fanfare')

    OpenAIRE

    Shyama Ranjani Weerakoon

    2010-01-01

    Direct and indirect somatic embryogenesis (SE) from petiole and leaf explants of Scaevola aemula R. Br. cv. 'Purple Fanfare' was achieved. High frequency of somatic embryos was obtained directly from petiole and leaf explants using an inductive plant growth regulator signal thidiazuron (TDZ). Petiole explants were more responsive to SE than leaves. Plants derived from somatic embryos of petiole explants germinated more readily into plants. SE occurred more efficiently in ...

  8. Plant regeneration via direct somatic embryogenesis from leaf explants of Tolumnia Louise Elmore 'Elsa'.

    Science.gov (United States)

    Shen, Hui-Ju; Chen, Jen-Tsung; Chung, Hsiao-Hang; Chang, Wei-Chin

    2018-01-22

    Tolumnia genus (equitant Oncidium) is a group of small orchids with vivid flower color. Thousands of hybrids have been registered on Royal Horticulture Society and showed great potential for ornamental plant market. The aim of this study is to establish an efficient method for in vitro propagation. Leaf explants taken from in vitro-grown plants were used to induce direct somatic embryogenesis on a modified 1/2 MS medium supplemented with five kinds of cytokinins, 2iP, BA, kinetin, TDZ and zeatin at 0.3, 1 and 3 mg l -1 in darkness. TDZ at 3 mg l -1 gave the highest percentage of explants with somatic globular embryos after 90 days of culture. It was found that 2,4-D and light regime highly retarded direct somatic embryogenesis and showed 95-100% of explant browning. Histological observations revealed that the leaf cells divided into meristematic cells firstly, followed by somatic proembryos, and then somatic globular embryos. Eventually, somatic embryos developed a bipolar structure with the shoot apical meristem and the root meristem. Scanning electron microscopy observations showed that the direct somatic embryogenesis from leaf explants was asynchronously. The somatic embryos were found on the leaf tip, the adaxial surface and also the mesophyll through a cleft, and it reflected the heterogeneity of the explant. The 90-day-old globular embryos were detached from the parent explants and transferred onto a hormone-free 1/2 MS medium in light condition for about 1 month to obtain 1-cm-height plantlets. After another 3 months for growth, the plantlets were potted with Sphagnum moss and were acclimatized in a shaded greenhouse. After 1 month of culture, the survival rate was 100%. In this report, a protocol for efficient regenerating a Tolumnia orchid, Louise Elmore 'Elsa', was established via direct somatic embryogenesis and might reveal an alternative approach for mass propagation of Tolumnia genus in orchid industry.

  9. A Novel 3D Skin Explant Model to Study Anaerobic Bacterial Infection

    Directory of Open Access Journals (Sweden)

    Grazieli Maboni

    2017-09-01

    Full Text Available Skin infection studies are often limited by financial and ethical constraints, and alternatives, such as monolayer cell culture, do not reflect many cellular processes limiting their application. For a more functional replacement, 3D skin culture models offer many advantages such as the maintenance of the tissue structure and the cell types present in the host environment. A 3D skin culture model can be set up using tissues acquired from surgical procedures or post slaughter, making it a cost effective and attractive alternative to animal experimentation. The majority of 3D culture models have been established for aerobic pathogens, but currently there are no models for anaerobic skin infections. Footrot is an anaerobic bacterial infection which affects the ovine interdigital skin causing a substantial animal welfare and financial impact worldwide. Dichelobacter nodosus is a Gram-negative anaerobic bacterium and the causative agent of footrot. The mechanism of infection and host immune response to D. nodosus is poorly understood. Here we present a novel 3D skin ex vivo model to study anaerobic bacterial infections using ovine skin explants infected with D. nodosus. Our results demonstrate that D. nodosus can invade the skin explant, and that altered expression of key inflammatory markers could be quantified in the culture media. The viability of explants was assessed by tissue integrity (histopathological features and cell death (DNA fragmentation over 76 h showing the model was stable for 28 h. D. nodosus was quantified in all infected skin explants by qPCR and the bacterium was visualized invading the epidermis by Fluorescent in situ Hybridization. Measurement of pro-inflammatory cytokines/chemokines in the culture media revealed that the explants released IL1β in response to bacteria. In contrast, levels of CXCL8 production were no different to mock-infected explants. The 3D skin model realistically simulates the interdigital skin and has

  10. ARG1 and ARL2 contribute to gravity signal transduction in the statocytes of Arabidopsis thaliana roots and hypocotyls

    Science.gov (United States)

    Masson, Patrick; Harrison, Benjamin; Stanga, John; Otegui, Marisa; Sedbrook, John

    Gravity is an important cue that plant organs use to guide their growth. Each organ is characterized by a defined gravity set point angle that dictates its optimal orientation within the gravity field. Specialized cells, named statocytes, enable this directional growth response by perceiving gravity via the sedimentation of, and/or tension/pressure exerted by, starch-filled plastids within their cytoplasm. Located in the columella region of the cap in roots and in the endodermis of hypocotyls and stems, these cells modulate the lateral transport of auxin across the corresponding organ in a gravistimulus-dependent manner. Upon plant reorientation within the gravity field, a gravity signal transduction pathway is activated within those cells, which in roots leads to a relocalization of the PIN3 auxin efflux carrier toward the lower membrane and an alkalinization of the cytoplasm. In turn, these events appear to promote a lateral transport of auxin toward the bottom side of the stimulated organ, which promotes a curvature. We previously uncovered ARG1 and ARL2 as essential contributors to these cellular processes. Mutations in these genes result in altered root and hypocotyl gravitropism. In roots, this abnormal growth behavior is associated with a lack of PIN3 relocalization within the statocytes and an absence of preferential downward auxin transport upon gravistimulation. These two genes encode paralogous J-domain proteins that are associated with the plasma membrane and other membranes of the vesicular trafficking pathway, and appear to modulate protein trafficking within the statocytes. An analysis of the root gravitropic phenotypes associated with different double mutant configurations affecting ARG1, ARL2 and PIN3 suggest that all three proteins function in a common gravity-signaling pathway. Surprisingly, when a mutation that affects starch biosynthesis (pgm) is introgressed into an arg1-2 mutant, the gravitropic defects are dramatically enhanced relative to

  11. Genotype, explant, medium, light and radiation effects on the in vitro plant regeneration in alfalfa (Medicago Sativa L.)

    International Nuclear Information System (INIS)

    El-Fiki, A.A.; Abdel-Hameed, A.A.M.; Sayed, A.I.H.

    2005-01-01

    The relative importance of genotype, explants, radiation, medium and light and their interactions for in vitro plant regeneration via somatic embryogenesis in alfalfa (Medicago sativa L.) has been studied. Shoot and leaf explants of two commercially grown Egyptian cultivars, Al-Wadi Al-Gadid and Siwa Tarkibi, were used in the study. The effect of gamma radiation doses 40, 80, 120 and 160 Gy were negative on plant regeneration, in spite of increase with some treatments. The best results of plant regeneration were obtained with dose 40 Gy with control light regime (16 h) on MS + 0.5 mg NAA + 1.5 mg BAP in both shoot and leaf explants of cv. Al-Wadi. The shoot explant of cv. Siwa was sensitive for gamma radiation dose 40 Gy while affirmative effect was obtained in leaf explant on MS + 1.0 mg NAA + 0.5 mg BAP with control light regime. However, dose 80 Gy showed the best results on MS + 0.5 mg NAA + 0.5 mg BAP in shoot and leaf explants of both cultivars, with control light regime in shoot explant and dark/light (DL) and dark/dark (DD) in leaf explant of cv. Al-Wadi, while with light/dark (LD) in shoot explant and control light regime in leaf explant of cv. Siwa. On the other hand, the highest plant regeneration ratio observed with dose 120 Gy were on 1.5 mg NAA + 0.5 mg BAP with control light regime in shoot and leaf explants of cv. Al-Wadi but on 0.5 mg NAA + 0.5 mg BAP with control and dark/light (DL) light regime in shoot and leaf explants of cv. Siwa. Whereas, the radiation dose 160 Gy showed severe effect on plant regeneration in both cultivars but highest percentage was observed on MS + 0.5 mg NAA + 0.5 mg BAP with dark/light (DL) in shoot explant, MS + 0.5 mg NAA + 1.5 mg BAP with control light regime in leaf explant of cv. Al-Wadi, MS + 0.5 mg NAA + 1.5 mg BAP in shoot explant and MS + 0.5 mg NAA + 0.5 mg BAP in leaf explant with dark/light (DL) in cv. Siwa. However, the effects of the same doses on callus growth showed that the highest callus weight was

  12. Multiplicação e enraizamento in vitro de amoreira-preta 'Xavante': efeito da concentração de sais, do tipo de explante e de carvão ativado no meio de cultura Blackberry 'Xavante' in vitro multiplication and rooting: salt concentrations effect, explant type, and activated coal on culture medium

    Directory of Open Access Journals (Sweden)

    Luciane Nolasco Leitzke

    2009-01-01

    concentrations in culture medium for. The first experiment constituted of nodal segments with 1 cm, originated from of the in vitro cultivation, inoculated at four different MS medium concentrations, supplemented with 7.5 µM BAP, 30 g L-1 sucrose, 100 mg L-1 myo-inositol and 6 g L-1 agar, and with two types of explants, which totaled 8 treatments. The second experiment constituted of apex microcutting, having about 1 to 1.5 cm length, and two leaves were inoculated at three different concentrations of MS medium, added of 30 g L-1 sucrose, 100 mg L-1 myo-inositol and 6 g L-1 agar and supplemented with three different activated coal concentrations, totaling nine treatments. It can be inferred that T1, in other words, explants with leaves, were more efficient at 125% MS salt concentration on culture medium, thereby inducing larger number of leaves, buds, shoots and shoot length, and that the use of 75% salt concentration without activated coal is the most suitable one for the blackberry 'Xavante' in vitro rooting.

  13. Osculum dynamics and filtration activity in small single-osculum explants of the demosponge Halichondria panicea

    DEFF Research Database (Denmark)

    Kumala, Lars; Riisgård, Hans Ulrik; Canfield, Donald Eugene

    2017-01-01

    the clearance method. Osculum dynamics, as expressed by temporal variation of the OSA, including osculum contraction and expansion, correlated with variability in the explant filtration rate, and no water pumping was observed during periods of osculum closure. A linear relationship between filtration rate (FR......Contraction-inflation behavior, including the closure and opening of the exhalant opening (osculum), is common among sponges. This behavior may temporally affect filtration activity, making it difficult to study and understand sponge feeding biology. To examine the interplay between osculum...... dynamics and filtration activity, small (18 mm3) single-osculum explants of the demosponge Halichondria panicea were studied. Time-lapse video stereo-microscope recordings of the osculum cross-sectional area (OSA) were made simultaneously with measurements of the filtration rate (∼15°C, ∼20 PSU) using...

  14. Acetylcholine causes rooting in leaf explants of in vitro raised tomato (Lycopersicon esculentum Miller) seedlings.

    Science.gov (United States)

    Bamel, Kiran; Gupta, Shrish Chandra; Gupta, Rajendra

    2007-05-30

    The animal neurotransmitter acetylcholine (ACh) induces rooting and promotes secondary root formation in leaf explants of tomato (Lycopersicon esculentum Miller var. Pusa Ruby), cultured in vitro on Murashige and Skoog's medium. The roots originate from the midrib of leaf explants and resemble taproot. ACh at 10(-5) M was found to be the optimum over a wide range of effective concentrations between 10(-7) and 10(-3) M. The breakdown products, choline and acetate were ineffective even at 10(-3) M concentration. ACh appears to have a natural role in tomato rhizogenesis because exogenous application of neostigmine, an inhibitor of ACh hydrolysis, could mimic the effect of ACh. Neostigmine, if applied in combination with ACh, potentiated the ACh effect.

  15. Aqueous extract of yellow maca (Lepidium meyenii) improves sperm count in experimental animals but response depends on hypocotyl size, pH and routes of administration.

    Science.gov (United States)

    Sanchez-Salazar, L; Gonzales, G F

    2018-04-01

    Lepidium meyenii, a Peruvian plant growing over 4000 m.a.s.l., has effects on nutrition and fertility. The purpose of this study was to evaluate the sperm count in 105 male mice receiving boiled aqueous extract of yellow maca hypocotyls from different sizes, under different pH conditions and using two different routes of administration. Five mice per group were treated daily for 3 days with vehicle (oral and intraperitoneal) or maca aqueous extracts (5 mg/0.5 ml/day) belonging to the first, second, third and fourth categories, according to their hypocotyl size. On day four, sperm count was evaluated at testis, epididymis and vas deferens. Sperm count was higher in mice receiving maca from the larger sizes (first and second categories). Reduction in maca extract pH increased sperm count, whereas an increase in the pH resulted in a reduction in sperm count. The effect of pH reduction is observed only in maca from the first and second categories. Aqueous extract of maca was effective only after oral administration. In conclusion, the larger size of hypocotyls presented the best biological effect, and the low pH in the extract and the transformation after gastrointestinal passage are both important for its biological action. © 2017 Blackwell Verlag GmbH.

  16. Clinicopathologic features of hepatic neoplasms in explanted livers: a single institution experience

    International Nuclear Information System (INIS)

    Mourad, W.; Tulbah, A.; Al-Omari, M.; Al-Mana, H.; Khalaf, H.; Neiamatallah, M.

    2007-01-01

    Hepatic neoplasms can be the primary indication for hepatic transplantation. The tumors can also be incidentally identified in explanted livers. We explored the clinicopathologic features of hepatic neoplasms identified in explanted livers. All explanted livers resected between 2001 and 2006 were evaluated for the presence of neoplasms and their clinicopathologic features were examined. In 198 liver transplants, 15 neoplasms (15.3%) were identified. Patient ages ranged from 5 to 63 years (median, 56 years). The primary etiology of hepatic disease was hepatitis C virus in 12 cases, hepatitis B virus in 1 case, cryptogenic cirrhosis in 1 case and congenital hepatic fibrosis in 1 case. Serum alpha-fetoprotein was significantly elevated (>400 U/L) in only 2 cases. CA19-9 was not elevated in any of the cases. The tumors included hepatocellular carcinoma (HCC) in 13 cases, 1 case of cholangiocarcinoma and 1 case of combined HCC and hepatoblastoma. The tumors ranged in size from 0.5 to 5 cm (median 1.4 cm) and were multifocal in 5 of the cases (33%). Tissue alpha-fetoprotein expression was only seen in the cases associated with elevated serum levels. In our institution hepatic neoplasma are seen in more than 15% of explanted livers. They can be incidentally identified, are frequently not associated with elevated serum levels of alpha-fetoprotein and CA19-9, are commonly multifocal but small and are associated with good prognosis. Elevated serum alpha-fetoprotein, albeit specific, is not a very sensitive marker in the detection of hepatic neoplasms. (author)

  17. E2f1 mediates high glucose-induced neuronal death in cultured mouse retinal explants.

    Science.gov (United States)

    Wang, Yujiao; Zhou, Yi; Xiao, Lirong; Zheng, Shijie; Yan, Naihong; Chen, Danian

    2017-10-02

    Diabetic retinopathy (DR) is the most common complication of diabetes and remains one of the major causes of blindness in the world; infants born to diabetic mothers have higher risk of developing retinopathy of prematurity (ROP). While hyperglycemia is a major risk factor, the molecular and cellular mechanisms underlying DR and diabetic ROP are poorly understood. To explore the consequences of retinal cells under high glucose, we cultured wild type or E2f1 -/- mouse retinal explants from postnatal day 8 with normal glucose, high osmotic or high glucose media. Explants were also incubated with cobalt chloride (CoCl 2 ) to mimic the hypoxic condition. We showed that, at 7 days post exposure to high glucose, retinal explants displayed elevated cell death, ectopic cell division and intact retinal vascular plexus. Cell death mainly occurred in excitatory neurons, such as ganglion and bipolar cells, which were also ectopically dividing. Many Müller glial cells reentered the cell cycle; some had irregular morphology or migrated to other layers. High glucose inhibited the hyperoxia-induced blood vessel regression of retinal explants. Moreover, inactivation of E2f1 rescued high glucose-induced ectopic division and cell death of retinal neurons, but not ectopic cell division of Müller glial cells and vascular phenotypes. This suggests that high glucose has direct but distinct effects on retinal neurons, glial cells and blood vessels, and that E2f1 mediates its effects on retinal neurons. These findings shed new light onto mechanisms of DR and the fetal retinal abnormalities associated with maternal diabetes, and suggest possible new therapeutic strategies.

  18. Candidate Microbicides Block HIV-1 Infection of Human Immature Langerhans Cells within Epithelial Tissue Explants

    Science.gov (United States)

    Kawamura, Tatsuyoshi; Cohen, Sandra S.; Borris, Debra L.; Aquilino, Elisabeth A.; Glushakova, Svetlana; Margolis, Leonid B.; Orenstein, Jan M.; Offord, Robin E.; Neurath, A. Robert; Blauvelt, Andrew

    2000-01-01

    Initial biologic events that underlie sexual transmission of HIV-1 are poorly understood. To model these events, we exposed human immature Langerhans cells (LCs) within epithelial tissue explants to two primary and two laboratory-adapted HIV-1 isolates. We detected HIV-1Ba-L infection in single LCs that spontaneously emigrated from explants by flow cytometry (median of infected LCs = 0.52%, range = 0.08–4.77%). HIV-1–infected LCs downregulated surface CD4 and CD83, whereas MHC class II, CD80, and CD86 were unchanged. For all HIV-1 strains tested, emigrated LCs were critical in establishing high levels of infection (0.1–1 μg HIV-1 p24 per milliliter) in cocultured autologous or allogeneic T cells. HIV-1Ba-L (an R5 HIV-1 strain) more efficiently infected LC–T cell cocultures when compared with HIV-1IIIB (an X4 HIV-1 strain). Interestingly, pretreatment of explants with either aminooxypentane-RANTES (regulated upon activation, normal T cell expressed and secreted) or cellulose acetate phthalate (potential microbicides) blocked HIV-1 infection of LCs and subsequent T cell infection in a dose-dependent manner. In summary, we document HIV-1 infection in single LCs after exposure to virus within epithelial tissue, demonstrate that relatively low numbers of these cells are capable of inducing high levels of infection in cocultured T cells, and provide a useful explant model for testing of agents designed to block sexual transmission of HIV-1. PMID:11085750

  19. Effects of glucosamine on proteoglycan loss by tendon, ligament and joint capsule explant cultures.

    Science.gov (United States)

    Ilic, M Z; Martinac, B; Samiric, T; Handley, C J

    2008-12-01

    To investigate the effect of glucosamine on the loss of newly synthesized radiolabeled large and small proteoglycans by bovine tendon, ligament and joint capsule. The kinetics of loss of (35)S-labeled large and small proteoglycans from explant cultures of tendon, ligament and joint capsule treated with 10mM glucosamine was investigated over a 10-day culture period. The kinetics of loss of (35)S-labeled small proteoglycans and the formation of free [(35)S]sulfate were determined for the last 10 days of a 15-day culture period. The proteoglycan core proteins were analyzed by gel electrophoresis followed by fluorography. The metabolism of tendon, ligament and joint capsule explants exposed to 10mM glucosamine was evaluated by incorporation of [(3)H]serine and [(35)S]sulfate into protein and glycosaminoglycans, respectively. Glucosamine at 10mM stimulated the loss of small proteoglycans from ligament explant cultures. This was due to the increased loss of both macromolecular and free [(35)S]sulfate to the medium indicating that glucosamine affected the release of small proteoglycans as well as their intracellular degradation. The degradation pattern of small proteoglycans in ligament was not affected by glucosamine. In contrast, glucosamine did not have an effect on the loss of large or small proteoglycans from tendon and joint capsule or large proteoglycans from ligament explant cultures. The metabolism of cells in tendon, ligament and joint capsule was not impaired by the presence of 10mM glucosamine. Glucosamine stimulated the loss of small proteoglycans from ligament but did not have an effect on small proteoglycan catabolism in joint capsule and tendon or large proteoglycan catabolism in ligament, tendon or synovial capsule. The consequences of glucosamine therapy at clinically relevant concentrations on proteoglycan catabolism in joint fibrous connective tissues need to be further assessed in an animal model.

  20. Effect of BAP (6-benzylaminopurine on shoot induction in explants of brazilwood

    Directory of Open Access Journals (Sweden)

    Ana Katarina Oliveira Aragão

    2011-09-01

    Full Text Available The Brazilian Atlantic forest has been subjected to intense degradation, with only about 7% to 8% of its original area remaining today. This situation has raised concerns over the conservation of species threatened with extinction. In all, 276 tree and bush species are under threat, out of which this study chose to evaluate alternatives for protecting brazilwood ‘Pau-Brasil’ (Caesalpinia echinata Lam.. Most studies performed so far on this subject either evaluate the effect of cytokinins on induction of callogenesis or focus on improving cryopreservation methodologies. In an attempt to expand knowledge about biotechnological techniques enabling conservation of C. echinata, this work evaluated the effect of 6-benzylaminopurine (BAP and explant type on induction of shoots in brazilwood. To attain that, explants were inoculated into basic MS medium and into MS medium supplemented with 2.5 µM, 3.5 µM and 4.5 µM of BAP, and kept in a growth room for 40 days under controlled photoperiod and temperature conditions. A 2x4 factorial design was adopted, with three replicates. Analyzed variables included shoot percentage, callogenesis and oxidations, and means were compared by the Tukey test at the 5% probability level. Results showed a significant influence of BAP only on shoot induction, and of explant type on that variable and on other variables too. It was concluded that, under in vitro conditions, the nodal type of explant is more responsive to BAP action and that 2.5 µM is the recommended concentration for shoot induction in brazilwood.

  1. Carboxylesterase-dependent cytotoxicity of dibasic esters (DBE) in rat nasal explants.

    Science.gov (United States)

    Trela, B A; Bogdanffy, M S

    1991-02-01

    Dibasic esters (DBE) are a solvent mixture of dimethyl adipate (DMA), dimethyl glutarate (DMG), and dimethyl succinate (DMS) used in the paint and coating industry. Subchronic inhalation toxicity studies have demonstrated that DBE induce a mild degeneration of the olfactory, but not the respiratory, epithelium of the rat nasal cavity. Carboxylesterase-mediated hydrolysis of the individual dibasic esters is more efficient in olfactory than in respiratory mucosal homogenates. In the present study, an in vitro system of cultured rat nasal explants was utilized to determine if DBE toxicity is dependent on a metabolic activation by nonspecific carboxylesterase. Explants from both the olfactory and the respiratory regions of the female rat nasal cavity were incubated for 2 hr in Williams' medium E containing 10-100 mM DMA, DMG, or DMS. DBE caused a dose-related increase in nasal explant acid phosphatase release, a biochemical index of cytotoxicity. HPLC analysis demonstrated parallel increases in the carboxylesterase-mediated formation of monomethyl ester metabolites. Diacid metabolite production in the nasal explant system was not entirely concentration-dependent. Metabolite concentrations and acid phosphatase release were generally greater in olfactory than respiratory tissues. DBE-induced cytotoxicity and acid metabolite production were markedly attenuated in nasal tissue excised from rats which were pretreated with bis(p-nitrophenyl)phosphate, a carboxylesterase inhibitor. This study presents a viable in vitro method for assessing organic ester cytotoxicity in the rat nasal cavity. It was shown that DBE are weak nasal toxicants under the conditions of this system. It was further demonstrated that DBE toxicity is dependent on a carboxylesterase-mediated activation. A similar mechanism was proposed for the nasal toxicity induced by other organic esters following inhalation exposure.

  2. In vitro maintenance of spermatogenesis in Xenopus laevis testis explants cultured in serum-free media

    International Nuclear Information System (INIS)

    Risley, M.S.; Miller, A.; Bumcrot, D.A.

    1987-01-01

    Spermatogenesis has been maintained for extended periods in Xenopus laevis testis explants cultured in serum-free media supplemented with bovine serum albumin, insulin, transferrin, follicle-stimulating hormone, dihydrotestosterone, testosterone, retinol, ascorbate, and tocopherol. The organization of the testis fragments was maintained for 28 days, and all stages of development were present throughout the culture period. 3 H-Thymidine-labeled secondary (Type B) spermatogonia developed in 28 days into spermatids at the acrosomal vesicle stage whereas labeled zygotene spermatocytes became mature spermatids in 28 days. Spermatogonial proliferation also continued in vitro for 28 days. Germ cell differentiation was not dependent upon exogenous testosterone, ascorbate, or tocopherol since 3 H-labeled spermatogonia became mature spermatids in testes cultured 35 days in media lacking these supplements. Autoradiography demonstrated that 55% of the luminal sperm present in explants cultured 10 days had differentiated in vitro. Sperm from testes cultured 10-35 days were similar to sperm from freshly dissected testes with regard to motility and fecundity, and eggs fertilized with sperm from explant cultures developed normally into swimming tadpoles. The results demonstrate the feasibility of maintaining vertebrate spermatogenesis in culture and suggest that in vitro analysis of Xenopus spermatogenesis using defined media may provide important insights into the evolution of regulatory mechanisms in spermatogenesis

  3. Phytic acid decreases deoxynivalenol and fumonisin B1-induced changes on swine jejunal explants

    Directory of Open Access Journals (Sweden)

    Elisângela Olegário da Silva

    2014-01-01

    Full Text Available The purpose of the present study was to investigate the effects of phytic acid (IP6 on morphological and immunohistochemical parameters on intestinal explants exposed to deoxynivalenol (DON and fumonisin B1 (FB1. The jejunal explants were exposed for 4 h to different treatments: control, DON (10 μM, DON plus 2.5 mM or 5 mM IP6, FB1 (70 μM, and FB1 plus 2.5 mM or 5 mM IP6. Both mycotoxins induced significant intestinal lesions and decreased villi height. The presence of 2.5 mM and 5 mM IP6 significantly inhibited the morphological changes caused by the mycotoxins. DON induced a significant increase in caspase-3 (83% and cyclooxygenase-2 (71.3% expression compared with the control. The presence of 5 mM IP6 induced a significant decrease in caspase-3 (43.7% and Cox-2 (48% expression compared with the DON group. FB1 induced a significant increase in caspase-3 expression (47% compared to the control, whereas IP6 induced no significant change in this expression. A significant decrease in cell proliferation was observed when explants were exposed to 5 mM of IP6 in comparison with the DON and FB1 groups. The present data provide evidence that phytic acid modulates the toxic effects induced by DON and FB1 on intestinal tissue.

  4. Pistacia lentiscus fruit oil reduces oxidative stress in human skin explants caused by hydrogen peroxide.

    Science.gov (United States)

    Ben Khedir, S; Moalla, D; Jardak, N; Mzid, M; Sahnoun, Z; Rebai, T

    2016-10-01

    We investigated the efficacy of Pistacia lentiscus fruit oil (PLFO) for protecting human skin from damage due to oxidative stress. PLFO contains natural antioxidants including polyphenols, sterols and tocopherols. We compared the antioxidant potential of PLFO with extra virgin olive oil (EVOO). Explants of healthy adult human skin were grown in culture with either PLFO or EVOO before adding hydrogen peroxide (H 2 O 2 ). We also used cultured skin explants to investigate the effects of PLFO on lipid oxidation and depletion of endogenous antioxidant defense enzymes including glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) one day after 2 h exposure to H 2 O 2 . We found that PLFO scavenged radicals and protected skin against oxidative injury. PLFO exhibited greater antioxidant and free radical scavenging activity than EVOO. Skin explants treated with PLFO inhibited H 2 O 2 induced MDA formation by inhibition of lipid oxidation. In addition, the oil inhibited H 2 O 2 induced depletion of antioxidant defense enzymes including GPx, SOD and CAT. We found that treatment with PLFO repaired skin damage owing to its antioxidant properties.

  5. Callogenesis in root explants of four species of the family Solanaceae after inducing by Agrobacterium rhizogenes

    Directory of Open Access Journals (Sweden)

    Zahra Shakeran

    2015-09-01

    Full Text Available Studying explants affected by Agrobacterium rhizogenes shows that in addition to possible formation of hairy roots, it is likely that callogenesis can be induced in these tissues. The T-DNA region of A. rhizogenes codes enzymes that participate in biosynthesis of plants growth hormones. These hormones also affect callogenesis, hence, the formation of various calluses with different morphological properties are possible. It is very likely that the level of biosynthetic growth hormone, the plasmid carried by each bacteria strain, the position of T-DNA, and the level of gene expression contribute to this morphologic variation. In this study, the root explants of four species of the family Solanaceae namely Atropa belladonna, Datura metel, D. stramonium and Hyoscyamus niger were induced by using different strains of A. rhizogenes (A4, A7, AR15834, AR318, AR9402 and AR9543. Some of these explants entered callus phase and formed various calluses with different colors and shapes. Moreover, in some callus samples hairy roots were also appeared. These variations were probably caused by variations in the levels and ratios of auxin and cytokinine hormons after the induction. As shown in previous studies, the amount of secondary metabolites is reduced due to undifferentiated tissue produced in the callogenesis process.

  6. Electron microscopic evaluation of a gold glaucoma micro shunt after explantation.

    Science.gov (United States)

    Berk, Thomas A; Tam, Diamond Y; Werner, Liliana; Mamalis, Nick; Ahmed, Iqbal Ike K

    2015-03-01

    We present a case of an explanted gold glaucoma micro shunt (GMS Plus) and the subsequent light and electron microscopic analyses. The shunt was implanted in a patient with medically refractive glaucoma. The intraocular pressure (IOP) was stable at 12 mm Hg 6 months postoperatively but spiked to 26 mm Hg 6 months later; membranous growth was visible on the implant gonioscopically. A second gold micro shunt was placed 2 years after the first. The IOP was 7 mm Hg 1 week postoperatively but increased to 23 mm Hg 3 weeks later; similar membranous growth was visible on this implant. One of the shunts was explanted, and light and scanning electron microscopic analyses revealed encapsulation around the shunt exterior and connective tissue invasion of the microstructure. This represents the first electron microscopic analysis of an explanted gold glaucoma micro shunt and the first unequivocal images of the fibrotic pseudo-capsule traversing its microchannels and fenestrations. Dr. Ahmed is a consultant to and has received research grants from Solx, Inc. No other author has a financial or proprietary interest in any material or method mentioned. Copyright © 2015 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

  7. Chemosensitivity and radiosensitivity testing of freshly explanted human tumour cells in vitro

    International Nuclear Information System (INIS)

    Wells, J.

    1977-10-01

    In this thesis, in vitro testing for the chemosensitivity and radiosensitivity of freshly explanted human tumour cells is described. The cells were incubated with anti-tumour drugs and either a 6-day growth test performed or a clonal growth test as a measure of survival of cell reproductive capacity. It was shown that if one aims to develop a suitable in vitro method for predicting the subsequent response of human tumour cells in situ to cytotoxic chemotherapy, the test procedure must be initiated before the explanted cells have undergone significant growth in vitro. The survival of the reproductive capacity of tumour cell explants following X-radiation was also studied. Using a 'feeder' layer technique, values for the survival curve parameter Dsub(q) were in the range 400-610 rad and the values for D 0 were in the range 120-160 rad. The shape of the X-ray survival curves did not change when cells were retested after repeated subculturing in vitro. Therefore, unlike chemosensitivity measured by the same biological end-point, radiosensitivity apparently does not change once cells have reached their maximum growth potential. (UK)

  8. In vitro regeneration from petiole explants of non-toxic Jatropha curcas

    KAUST Repository

    Kumar, Nitish

    2011-01-01

    Jatropha curcas, a multipurpose shrub has acquired significant economic potential as biodiesel plant. The seeds or pressed cake is toxic due to the presence of toxic substances and is not useful as food/fodder despite having the best protein composition. A simple, efficient, and reproducible method for plant regeneration through direct organogenesis from petiole explants of non-toxic J. curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (57.61%), and number of shoot buds (4.98) per explant were obtained when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 mu M TDZ. The Induced shoot buds were transferred to MS medium containing 10 mu M kinetin (Kn), 4.5 mu M 6-benzyl aminopurine (BA), and 5.5 mu M alpha-naphthaleneacetic acid (NAA) for shoot proliferation and subsequent elongation was achieved on MS medium supplemented with 2.25 mu M BA and 8.5 mu M IAA. The elongated shoots could be rooted on half-strength MS medium with 15 mu M IBA, 11.4 mu M IAA and 5.5 mu M NAA with more than 90% survival rate. (C) 2010 Elsevier B.V. All rights reserved.

  9. Differential antiepileptic effects of the organic calcium antagonists verapamil and flunarizine in neurons of organotypic neocortical explants from newborn rats

    NARCIS (Netherlands)

    Bingmann, D; Speckmann, E J; Baker, R E; Ruijter, J; de Jong, B. M.

    1988-01-01

    Effects of the organic calcium antagonists verapamil and flunarizine on pentylenetetrazol induced paroxysmal depolarizations were tested in organotypic neocortical explants taken from neonatal rats. In these in vitro experiments the papaverin derivative verapamil depressed, and finally abolished,

  10. The parasitic plant Cuscuta australis is highly insensitive to abscisic acid-induced suppression of hypocotyl elongation and seed germination.

    Science.gov (United States)

    Li, Juan; Hettenhausen, Christian; Sun, Guiling; Zhuang, Huifu; Li, Jian-Hong; Wu, Jianqiang

    2015-01-01

    Around 1% of angiosperms are parasitic plants. Their growth and development solely or partly depend on host plants from which they extract water, nutrients, and other molecules using a parasitic plant-specific organ, the haustorium. Strong depletion of nutrients can result in serious growth retardation and in some cases, death of the hosts. The genus Cuscuta (dodder) comprises about 200 holoparasitic species occurring on all continents. Their seedlings have no roots and cotyledons but are only string-like hypocotyls. When they contact suitable host plants, haustoria are formed and thereafter seedlings rapidly develop into vigorously growing branches without roots and leaves. This highly specialized lifestyle suggests that Cuscuta plants likely have unique physiology in development and stress responses. Using germination and seedling growth assays, we show that C. australis seeds and seedlings are highly insensitive to abscisic acid (ABA). Transcriptome analysis and protein sequence alignment with Arabidopsis, tomato, and rice homologs revealed that C. australis most likely consists of only four functional ABA receptors. Given that Cuscuta plants are no longer severely challenged by drought stress, we hypothesize that the ABA-mediated drought resistance pathway in Cuscuta spp. might have had degenerated over time during evolution.

  11. The parasitic plant Cuscuta australis is highly insensitive to abscisic acid-induced suppression of hypocotyl elongation and seed germination.

    Directory of Open Access Journals (Sweden)

    Juan Li

    Full Text Available Around 1% of angiosperms are parasitic plants. Their growth and development solely or partly depend on host plants from which they extract water, nutrients, and other molecules using a parasitic plant-specific organ, the haustorium. Strong depletion of nutrients can result in serious growth retardation and in some cases, death of the hosts. The genus Cuscuta (dodder comprises about 200 holoparasitic species occurring on all continents. Their seedlings have no roots and cotyledons but are only string-like hypocotyls. When they contact suitable host plants, haustoria are formed and thereafter seedlings rapidly develop into vigorously growing branches without roots and leaves. This highly specialized lifestyle suggests that Cuscuta plants likely have unique physiology in development and stress responses. Using germination and seedling growth assays, we show that C. australis seeds and seedlings are highly insensitive to abscisic acid (ABA. Transcriptome analysis and protein sequence alignment with Arabidopsis, tomato, and rice homologs revealed that C. australis most likely consists of only four functional ABA receptors. Given that Cuscuta plants are no longer severely challenged by drought stress, we hypothesize that the ABA-mediated drought resistance pathway in Cuscuta spp. might have had degenerated over time during evolution.

  12. Clathrin-Mediated Auxin Efflux and Maxima Regulate Hypocotyl Hook Formation and Light-Stimulated Hook Opening in Arabidopsis.

    Science.gov (United States)

    Yu, Qinqin; Zhang, Ying; Wang, Juan; Yan, Xu; Wang, Chao; Xu, Jian; Pan, Jianwei

    2016-01-04

    The establishment of auxin maxima by PIN-FORMED 3 (PIN3)- and AUXIN RESISTANT 1/LIKE AUX1 (LAX) 3 (AUX1/LAX3)-mediated auxin transport is essential for hook formation in Arabidopsis hypocotyls. Until now, however, the underlying regulatory mechanism has remained poorly understood. Here, we show that loss of function of clathrin light chain CLC2 and CLC3 genes enhanced auxin maxima and thereby hook curvature, alleviated the inhibitory effect of auxin overproduction on auxin maxima and hook curvature, and delayed blue light-stimulated auxin maxima reduction and hook opening. Moreover, pharmacological experiments revealed that auxin maxima formation and hook curvature in clc2 clc3 were sensitive to auxin efflux inhibitors 1-naphthylphthalamic acid and 2,3,5-triiodobenzoic acid but not to the auxin influx inhibitor 1-naphthoxyacetic acid. Live-cell imaging analysis further uncovered that loss of CLC2 and CLC3 function impaired PIN3 endocytosis and promoted its lateralization in the cortical cells but did not affect AUX1 localization. Taken together, these results suggest that clathrin regulates auxin maxima and thereby hook formation through modulating PIN3 localization and auxin efflux, providing a novel mechanism that integrates developmental signals and environmental cues to regulate plant skotomorphogenesis and photomorphogenesis. Copyright © 2016 The Author. Published by Elsevier Inc. All rights reserved.

  13. The Pattern of Distribution of Peroxidase and H2O2 in Hypocotyls of Pumpkin irradiated with Gamma Ray

    International Nuclear Information System (INIS)

    Wi, Seung Gon; Chung, Byung Yeoup; Kim, Jae Sung; Kim, Jin Hong; Lee, Ju Woon; Baek, Myung Hwa; Chae, Hyo Seok

    2005-01-01

    Gamma radiation, one of ionizing radiations, has been reported to affect the morphological, anatomical, biochemical and physiological changes of plants at different dose levels. These effects at high level include inhibition in plant growth. Peroxidases (PODs) are mainly participations the process of lignification on the cell wall and protect the cell organelles in cytosol against the oxidative stresses by ROS (reactive oxygen species). Hydrogen peroxide (ROS) is normal metabolite in aerobic cells and the physiological steady concentrations (between 10 -7 and 10 -9 ) are not particularly cytotoxic. When these concentrations are increased by ionizing radiation, they lead to cell lethality. Thus radiation-induced H 2 O 2 may appear as an important agent causing cell damage. In this study, a polyclonal antibody against peroxidase and cerium chloride as a trapping agent for H 2 O 2 were used to obtain for better information on the occurrence and distribution of POD and H 2 O 2 in the cytoplasm and walls of vascular bundle in hypocotyls of pumpkin

  14. Segmented trapped vortex cavity

    Science.gov (United States)

    Grammel, Jr., Leonard Paul (Inventor); Pennekamp, David Lance (Inventor); Winslow, Jr., Ralph Henry (Inventor)

    2010-01-01

    An annular trapped vortex cavity assembly segment comprising includes a cavity forward wall, a cavity aft wall, and a cavity radially outer wall there between defining a cavity segment therein. A cavity opening extends between the forward and aft walls at a radially inner end of the assembly segment. Radially spaced apart pluralities of air injection first and second holes extend through the forward and aft walls respectively. The segment may include first and second expansion joint features at distal first and second ends respectively of the segment. The segment may include a forward subcomponent including the cavity forward wall attached to an aft subcomponent including the cavity aft wall. The forward and aft subcomponents include forward and aft portions of the cavity radially outer wall respectively. A ring of the segments may be circumferentially disposed about an axis to form an annular segmented vortex cavity assembly.

  15. Pavement management segment consolidation

    Science.gov (United States)

    1998-01-01

    Dividing roads into "homogeneous" segments has been a major problem for all areas of highway engineering. SDDOT uses Deighton Associates Limited software, dTIMS, to analyze life-cycle costs for various rehabilitation strategies on each segment of roa...

  16. Speaker segmentation and clustering

    OpenAIRE

    Kotti, M; Moschou, V; Kotropoulos, C

    2008-01-01

    07.08.13 KB. Ok to add the accepted version to Spiral, Elsevier says ok whlile mandate not enforced. This survey focuses on two challenging speech processing topics, namely: speaker segmentation and speaker clustering. Speaker segmentation aims at finding speaker change points in an audio stream, whereas speaker clustering aims at grouping speech segments based on speaker characteristics. Model-based, metric-based, and hybrid speaker segmentation algorithms are reviewed. Concerning speaker...

  17. Spinal segmental dysgenesis

    Directory of Open Access Journals (Sweden)

    N Mahomed

    2009-06-01

    Full Text Available Spinal segmental dysgenesis is a rare congenital spinal abnormality , seen in neonates and infants in which a segment of the spine and spinal cord fails to develop normally . The condition is segmental with normal vertebrae above and below the malformation. This condition is commonly associated with various abnormalities that affect the heart, genitourinary, gastrointestinal tract and skeletal system. We report two cases of spinal segmental dysgenesis and the associated abnormalities.

  18. Automatic Melody Segmentation

    NARCIS (Netherlands)

    Rodríguez López, Marcelo

    2016-01-01

    The work presented in this dissertation investigates music segmentation. In the field of Musicology, segmentation refers to a score analysis technique, whereby notated pieces or passages of these pieces are divided into “units” referred to as sections, periods, phrases, and so on. Segmentation

  19. Elongated Hypocotyl 5-Homolog (HYH Negatively Regulates Expression of the Ambient Temperature-Responsive MicroRNA Gene MIR169

    Directory of Open Access Journals (Sweden)

    Phanu T. Serivichyaswat

    2017-12-01

    Full Text Available Arabidopsis microRNA169 (miR169 is an ambient temperature-responsive microRNA that plays an important role in stress responses and the floral transition. However, the transcription factors that regulate the expression of MIR169 have remained unknown. In this study, we show that Elongated Hypocotyl 5-Homolog (HYH directly binds to the promoter of MIR169a and negatively regulates its expression. Absolute quantification identified MIR169a as the major locus producing miR169. GUS reporter assays revealed that the deletion of a 498-bp fragment (–1,505 to –1,007, relative to the major transcriptional start site of MIR169a abolished its ambient temperature-responsive expression. DNA-affinity chromatography followed by liquid chromatography-mass spectrometry analysis identified transcription factor HYH as a trans-acting factor that binds to the 498-bp promoter fragment of pri-miR169a. Electrophoretic mobility shift assays and chromatin immunoprecipitation–quantitative PCR demonstrated that the HYH.2 protein, a predominant isoform of HYH, directly associated with a G-box-like motif in the 498-bp fragment of pri-miR169a. Higher enrichment of HYH.2 protein on the promoter region of MIR169a was seen at 23°C, consistent with the presence of more HYH.2 protein in the cell at the temperature. Transcript levels of pri-miR169a increased in hyh mutants and decreased in transgenic plants overexpressing HYH. Consistent with the negative regulation of MIR169a by HYH, the diurnal levels of HYH mRNA and pri-miR169a showed opposite patterns. Taken together, our results suggest that HYH is a transcription factor that binds to a G-box-like motif in the MIR169a promoter and negatively regulates ambient temperature-responsive expression of MIR169a at higher temperatures in Arabidopsis.

  20. In vitro establishment of nodal segments of Annona muricata L. young plants

    Directory of Open Access Journals (Sweden)

    Leyanis García-Águila

    2012-10-01

    Full Text Available The use of tissue in vitro culture for plant propagation of soursop (Annona muricata L. promissory trees can help increasing the availability of plants for developing field plantations. Considering these aspect, this work aimed to establish in vitro nodal segments of young plants of soursop. Nodal segments with 1.5 cm of length were superficially disinfected with 70% ethanol during one minute and with sodium hypochlorite (0.5, 1.0, 1.5 and 2.0% during 15 minutes. The presence of microbial contamination and the number of segments with axillary buds were evaluated after 15 and 25 days of culture. The length (cm of buds was also determined. Results showed a low incidence of microbial contamination in the explants because the presence of fungi in treatments was not observed. However, 3.8% segments were contaminated with bacteria in the treatment with lower concentration of sodium hypochlorite. Axillary shoots were observed in 73.0% of explants when 1.0% of sodium hypochlorite was used, without significant differences using 1.5%. Shoots development with first expanded leaves and a length ranged between 0.8 and 1.5 cm was observed after 25 days of culture. Increasing culture time, plants showed leaf abscission. These results demonstrate that in vitro culture can be used for the propagation of soursop. However, we must make emphasis in the study of the culture conditions for the multiplication phase. Key words: in vitro culture, microbial contamination, phenolic oxidation, shoot production.

  1. Effects of thidiazuron and paclobutrazol on regeneration potential of tulip flower stalk explants in vitro and subsequent shoot multiplication

    Directory of Open Access Journals (Sweden)

    Małgorzata Podwyszyńska

    2011-01-01

    Full Text Available The effects of TDZ and paclobutrazol on the primary regeneration on tulip flower stalk explants of six cultivars and subsequent shoot multiplication were examined. Explants, flower stalk slices, were excised from cooled and subsequently forced bulbs. The explants were incubated for two months in darkness on medium containing NAA and cytokinins, 2iP and BAP, as control, or TDZ (0.5-4 mg l-1 and paclobutrazol (0.05-0.4 mg l-1. Then, the regenerating explants were subcultured on medium with TDZ and NAA applied at low concentrations. Different regeneration capabilities were found depending on cultivar and growth regulators. The percentage of explants forming leaf-like structures ranged, on the control medium, from 80% in 'Blue Parrot' and 'Prominence' to below 30% in 'Apeldoorn' and 'Mirjoran'. TDZ, applied at optimum for each cultivar concentration, greatly increased the regeneration potential up to 70-100%. Paclobutrazol, added to the TDZ-containing medium, significantly enhanced the response of explants, resulting in high numbers of leaf-like structures formed per explant (13.7-22.8. The structures developed gradually into characteristic forms: the growing up cotyledonary leaf, the probable root primordium formed at its base, the growing downwards stolon and the shoot meristem developed finely on its tip. It is suggested that such primary regeneration may have a nature of somatic embryogenesis. Then, the adventitious shoots developed and formed clusters, which were divided into 2-3 smaller ones every two months. The growth regulators, used at initial stage, markedly influenced subsequent shoot multiplication. Thus, the most intensive shoot formation was noted with TDZ at concentrations of 0.5-2 mg l-1 and paclobutrazol of 0.05-0.1 mg l-1.

  2. Neuronal differentiation of hair-follicle-bulge-derived stem cells co-cultured with mouse cochlear modiolus explants.

    Directory of Open Access Journals (Sweden)

    Timo Schomann

    Full Text Available Stem-cell-based repair of auditory neurons may represent an attractive therapeutic option to restore sensorineural hearing loss. Hair-follicle-bulge-derived stem cells (HFBSCs are promising candidates for this type of therapy, because they (1 have migratory properties, enabling migration after transplantation, (2 can differentiate into sensory neurons and glial cells, and (3 can easily be harvested in relatively high numbers. However, HFBSCs have never been used for this purpose. We hypothesized that HFBSCs can be used for cell-based repair of the auditory nerve and we have examined their migration and incorporation into cochlear modiolus explants and their subsequent differentiation. Modiolus explants obtained from adult wild-type mice were cultured in the presence of EF1α-copGFP-transduced HFBSCs, constitutively expressing copepod green fluorescent protein (copGFP. Also, modiolus explants without hair cells were co-cultured with DCX-copGFP-transduced HFBSCs, which demonstrate copGFP upon doublecortin expression during neuronal differentiation. Velocity of HFBSC migration towards modiolus explants was calculated, and after two weeks, co-cultures were fixed and processed for immunohistochemical staining. EF1α-copGFP HFBSC migration velocity was fast: 80.5 ± 6.1 μm/h. After arrival in the explant, the cells formed a fascicular pattern and changed their phenotype into an ATOH1-positive neuronal cell type. DCX-copGFP HFBSCs became green-fluorescent after integration into the explants, confirming neuronal differentiation of the cells. These results show that HFBSC-derived neuronal progenitors are migratory and can integrate into cochlear modiolus explants, while adapting their phenotype depending on this micro-environment. Thus, HFBSCs show potential to be employed in cell-based therapies for auditory nerve repair.

  3. Analysis of causes of intraocular lens explantations in the material of Department of Ophthalmology, Medical University of Lodz.

    Science.gov (United States)

    Wilczyński, Michał; Wilczyńska, Olena; Omulecki, Wojciech

    2009-01-01

    Implantation of intraocular lenses (IOLS) has become a standard practice in cataract surgery, however, similar to any other type of surgery, using IOLs is not complication-free and sometimes explantation of intraocular lenses may be necessary. This study was to gather data and analyze causes of intraocular lens explantations, performed in the Department of Ophthalmology, Medical University of Łódź. The data were gathered from medical documentation of all patients who underwent intraocular lens removal from January 2003 to July 2006. The examined group consisted of 16 patients (16 eyes): 9 women (fraction 0.56), and 7 men (fraction 0.44), at the age from 21 to 82 years (mean age 62.4 years, SD +/- 15.5). In all patients IOL explantation was performed under local, peribulbar anaesthesia. Two groups of patients were distinguished: patients who had an anterior chamber lens explanted (3 patients, fraction 0.19) and patients who underwent posterior chamber lens explantation (13 patients, fraction 0.81). Causes of AC IOL explantations were: vaulting of the IOL (1 eye, fraction 0.06), luxation of the IOL to the vitreous cavity (1 eye, fraction 0.06), and painful eyeball after anterior chamber lens implantation (1 eye, fraction 0.06). Causes of PC IOL explantations were: subluxation of the IOL (6 eyes, fraction 0.38), luxation of the lens to the vitreous cavity (3 eyes, fraction 0.19), luxation of the lens to the anterior chamber (1 eye, fraction 0.06), endophthalmitis (2 eyes, fraction 0.13) and incorrect lens power (1 eye, fraction 0.06). In the majority of eyes (n = 13, fraction 0.81) the removed implant was replaced by another intraocular lens, but 3 eyes (fraction 0.19) were left aphakic. We did not observe serious intra- or early postoperative complications which might influence the final result of the operation.

  4. Agrobacterium-mediated transformation of finger millet (Eleusine coracana (L.) Gaertn.) using shoot apex explants.

    Science.gov (United States)

    Ceasar, S Antony; Ignacimuthu, S

    2011-09-01

    A new Agrobacterium-mediated transformation system was developed for finger millet using shoot apex explants. The Agrobacterium strain LBA4404 harboring binary vector pCAMBIA1301, which contained hygromycin phosphotransferase (hptII) as selectable marker gene and β-glucuronidase (GUS) as reporter gene, was used for optimization of transformation conditions. Two finger millet genotypes, GPU 45 and CO 14, were used in this study. The optimal conditions for the Agrobacterium-mediated transformation of finger millet were found to be the co-cultivation of explants obtained on the 16th day after callus induction (DACI), exposure of explants for 30 min to agrobacterial inoculum and 3 days of co-cultivation on filter paper placed on medium supplemented with 100 μM acetosyringone (AS). Addition of 100 μM L: -cysteine in the selection medium enhanced the frequency of transformation and transgenic plant recovery. Both finger millet genotypes were transformed by Agrobacterium. A frequency of 19% transient expression with 3.8% stable transformation was achieved in genotype GPU 45 using optimal conditions. Five stably transformed plants were fully characterized by Southern blot analysis. A segregation analysis was also performed in four R(1) progenies, which showed normal Mendelian pattern of transgene segregation. The inheritance of transgenes in R(1) progenies was also confirmed by Southern blot analysis. This is the first report on Agrobacterium-mediated transformation of finger millet. This study underpins the introduction of numerous agronomically important genes into the genome of finger millet in the future.

  5. Efficient regeneration of sorghum, Sorghum bicolor (L.) Moench, from shoot-tip explant.

    Science.gov (United States)

    Syamala, D; Devi, Prathibha

    2003-12-01

    Novel protocols for production of multiple shoot-tip clumps and somatic embryos of Sorghum bicolor (L.) Moench were developed with long-term goal of crop improvement through genetic transformation. Multiple shoot-tip clumps were developed in vitro from shoot-tip explant of one-week old seedling, cultured on MS medium containing only BA (0.5, 1 or 2 mg/l) or both BA (1 or 2 mg/l) and 2,4-D (0.5 mg/l) with bi-weekly subculture. Somatic embryos were directly produced on the enlarged dome shaped growing structures that developed from the shoot-tips of one-week old seedling explants (without any callus formation) when cultured on MS medium supplemented with both 2,4-D (0.5 mg/l) and BA (0.5 mg/l). However, the supplementation of MS medium with only 2,4-D (0.5 mg/l) induced compact callus without any plantlet regeneration. Each multiple shoot-clump was capable of regenerating more than 80 shoots via an intensive differentiation of both axillary and adventitious shoot buds, the somatic embryos were capable of 90% germination, plant conversion and regeneration. The regenerated shoots could be efficiently rooted on MS medium containing indole-3-butyric acid (IBA 1 mg/l). The plants were successfully transplanted to glasshouse and grown to maturity with a survival rate of 98%. Morphogenetic response of the explants was found to be genotypically independent.

  6. A MIV-150/zinc acetate gel inhibits SHIV-RT infection in macaque vaginal explants.

    Science.gov (United States)

    Barnable, Patrick; Calenda, Giulia; Ouattara, Louise; Gettie, Agegnehu; Blanchard, James; Jean-Pierre, Ninochka; Kizima, Larisa; Rodríguez, Aixa; Abraham, Ciby; Menon, Radhika; Seidor, Samantha; Cooney, Michael L; Roberts, Kevin D; Sperling, Rhoda; Piatak, Michael; Lifson, Jeffrey D; Fernandez-Romero, Jose A; Zydowsky, Thomas M; Robbiani, Melissa; Teleshova, Natalia

    2014-01-01

    To extend our observations that single or repeated application of a gel containing the NNRTI MIV-150 (M) and zinc acetate dihydrate (ZA) in carrageenan (CG) (MZC) inhibits vaginal transmission of simian/human immunodeficiency virus (SHIV)-RT in macaques, we evaluated safety and anti-SHIV-RT activity of MZC and related gel formulations ex vivo in macaque mucosal explants. In addition, safety was further evaluated in human ectocervical explants. The gels did not induce mucosal toxicity. A single ex vivo exposure to diluted MZC (1∶30, 1∶100) and MC (1∶30, the only dilution tested), but not to ZC gel, up to 4 days prior to viral challenge, significantly inhibited SHIV-RT infection in macaque vaginal mucosa. MZC's activity was not affected by seminal plasma. The antiviral activity of unformulated MIV-150 was not enhanced in the presence of ZA, suggesting that the antiviral activity of MZC was mediated predominantly by MIV-150. In vivo administration of MZC and CG significantly inhibited ex vivo SHIV-RT infection (51-62% inhibition relative to baselines) of vaginal (but not cervical) mucosa collected 24 h post last gel exposure, indicating barrier effect of CG. Although the inhibitory effect of MZC (65-74%) did not significantly differ from CG (32-45%), it was within the range of protection (∼75%) against vaginal SHIV-RT challenge 24 h after gel dosing. Overall, the data suggest that evaluation of candidate microbicides in macaque explants can inform macaque efficacy and clinical studies design. The data support advancing MZC gel for clinical evaluation.

  7. Glucocorticoids affect 24 h clock genes expression in human adipose tissue explant cultures.

    Directory of Open Access Journals (Sweden)

    Purificación Gómez-Abellán

    Full Text Available to examine firstly whether CLOCK exhibits a circadian expression in human visceral (V and subcutaneous (S adipose tissue (AT in vitro as compared with BMAL1 and PER2, and secondly to investigate the possible effect of the glucocorticoid analogue dexamethasone (DEX on positive and negative clock genes expression.VAT and SAT biopsies were obtained from morbid obese women (body mass index ≥ 40 kg/m(2 (n = 6. In order to investigate rhythmic expression pattern of clock genes and the effect of DEX on CLOCK, PER2 and BMAL1 expression, control AT (without DEX and AT explants treated with DEX (2 hours were cultured during 24 h and gene expression was analyzed at the following times: 10:00 h, 14:00 h, 18:00 h, 22:00 h, 02:00 h and 06:00 h, using qRT-PCR.CLOCK, BMAL1 and PER2 expression exhibited circadian patterns in both VAT and SAT explants that were adjusted to a typical 24 h sinusoidal curve. PER2 expression (negative element was in antiphase with respect to CLOCK and in phase with BMAL1 expression (both positive elements in the SAT (situation not present in VAT. A marked effect of DEX exposure on both positive and negative clock genes expression patterns was observed. Indeed, DEX treatment modified the rhythmicity pattern towards altered patterns with a period lower than 24 hours in all genes and in both tissues.24 h patterns in CLOCK and BMAL1 (positive clock elements and PER2 (negative element mRNA levels were observed in human adipose explants. These patterns were altered by dexamethasone exposure.

  8. Implantable cardioverter-defibrillator explantation for overdiagnosed or overtreated congenital long QT syndrome.

    Science.gov (United States)

    Gaba, Prakriti; Bos, J Martijn; Cannon, Bryan C; Cha, Yong-Mei; Friedman, Paul A; Asirvatham, Samuel J; Ackerman, Michael J

    2016-04-01

    Primary treatment of long QT syndrome (LQTS) currently consists of beta-blocker therapy, although an implantable cardioverter-defibrillator (ICD) is considered for high-risk patients. However, both overdiagnosis and overtreatment must be avoided because their sequelae can be significant. The purpose of this study was to evaluate the prevalence and details of ICD explants in a cohort of patients from a tertiary genetic heart rhythm clinic for a previously rendered diagnosis of LQTS. Overall, 1227 consecutive patients were included in the study. All patients had been referred to the Mayo Clinic for evaluation of possible LQTS and subsequently were either diagnosed with LQTS or dismissed as normal. Further stratification of patients was conducted to assess how many patients had an ICD and how many had a subsequent ICD explant. In total, 170 patients (14%) had an ICD, including 157 of 670 patients (23%) with confirmed LQTS and 13 of 557 patients (2%) who did not have LQTS. Among these, 12 of 1227 (1%) had the ICD removed: 7 of 157 LQTS patients (4.5%) compared to 5 of 14 non-LQTS patients (36%). Before explant, 5 of 12 patients (42%) experienced inappropriate shocks, ranging from 2 to as many as 54 shocks. In addition, 4 had a device-related infection, and 9 had device malfunction (including lead dysfunction or fracture). None of these patients had a breakthrough cardiac event since removal of their ICD during 5.5 ± 3.5 years of follow-up. Implications of overdiagnosis and overtreatment are profound because unnecessary ICD placement can be associated with infection, malfunction, inappropriate shocks, and subsequent anxiety. Copyright © 2016 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.

  9. Characterization of TLR-induced inflammatory responses in COPD and control lung tissue explants

    Directory of Open Access Journals (Sweden)

    Pomerenke A

    2016-09-01

    Full Text Available Anna Pomerenke,1 Simon R Lea,1 Sarah Herrick,2 Mark A Lindsay,3 Dave Singh1 1Centre for Respiratory Medicine and Allergy, Institute of Inflammation and Repair, Manchester Academic Health Science Centre, The University of Manchester and University Hospital of South Manchester, NHS Foundation Trust, 2Institute of Inflammation and Repair, Manchester Academic Health Science Centre, University of Manchester, Manchester, 3Department of Pharmacy and Pharmacology, University of Bath, Bath, UK Purpose: Viruses are a common cause of exacerbations in chronic obstructive pulmonary disease (COPD. They activate toll-like receptors (TLRs 3, 7, and 8, leading to a pro-inflammatory response. We have characterized the responses of TLR3 and TLR7/8 in lung tissue explants from COPD patients and control smokers.Methods: We prepared lung whole tissue explants (WTEs from patients undergoing surgery for confirmed or suspected lung cancer. In order to mimic the conditions of viral infection, we used poly(I:C for TLR3 stimulation and R848 for TLR7/8 stimulation. These TLR ligands were used alone and in combination. The effects of tumor necrosis factor α (TNFα neutralization and dexamethasone on TLR responses were examined. Inflammatory cytokine release was measured by enzyme-linked immunosorbent assay and gene expression by quantitative real-time polymerase chain reaction.Results: WTEs from COPD patients released higher levels of pro-inflammatory cytokines compared with WTEs from smokers. Activation of multiple TLRs led to a greater than additive release of TNFα and CCL5. TNFα neutralization and dexamethasone treatment decreased cytokine release.Conclusion: This WTE model shows an enhanced response of COPD compared with controls, suggesting an increased response to viral infection. There was amplification of innate immune responses with multiple TLR stimulation. Keywords: COPD, poly(I:C, R848, cytokines, lung explant

  10. A MIV-150/zinc acetate gel inhibits SHIV-RT infection in macaque vaginal explants.

    Directory of Open Access Journals (Sweden)

    Patrick Barnable

    Full Text Available To extend our observations that single or repeated application of a gel containing the NNRTI MIV-150 (M and zinc acetate dihydrate (ZA in carrageenan (CG (MZC inhibits vaginal transmission of simian/human immunodeficiency virus (SHIV-RT in macaques, we evaluated safety and anti-SHIV-RT activity of MZC and related gel formulations ex vivo in macaque mucosal explants. In addition, safety was further evaluated in human ectocervical explants. The gels did not induce mucosal toxicity. A single ex vivo exposure to diluted MZC (1∶30, 1∶100 and MC (1∶30, the only dilution tested, but not to ZC gel, up to 4 days prior to viral challenge, significantly inhibited SHIV-RT infection in macaque vaginal mucosa. MZC's activity was not affected by seminal plasma. The antiviral activity of unformulated MIV-150 was not enhanced in the presence of ZA, suggesting that the antiviral activity of MZC was mediated predominantly by MIV-150. In vivo administration of MZC and CG significantly inhibited ex vivo SHIV-RT infection (51-62% inhibition relative to baselines of vaginal (but not cervical mucosa collected 24 h post last gel exposure, indicating barrier effect of CG. Although the inhibitory effect of MZC (65-74% did not significantly differ from CG (32-45%, it was within the range of protection (∼75% against vaginal SHIV-RT challenge 24 h after gel dosing. Overall, the data suggest that evaluation of candidate microbicides in macaque explants can inform macaque efficacy and clinical studies design. The data support advancing MZC gel for clinical evaluation.

  11. Micropropagation of Ficus religiosa L. via leaf explants and comparative evaluation of acetylcholinesterase inhibitory activity in the micropropagated and conventionally grown plants.

    Science.gov (United States)

    Siwach, Priyanka; Gill, Anita Rani

    2014-10-01

    A high-frequency, season-independent, in vitro regeneration of Ficusreligiosa was developed, followed by comparative acetylcholinesterase inhibitory (AChEI) activity assay of the in vitro raised and conventionally grown plants. The use of AChEI activity is the most accepted strategy for the treatment of Alzheimer disease. Fully expanded, mature leaves were cut into different segments to initiate the cultures. The middle section of the leaf in vertical orientation with cut portion inserted inside the medium was found most suitable for direct shoot regeneration. Leaf explants responded with nearly consistent frequency (60-66.67 %) throughout the year. To obtain high frequency response with enhanced shoot multiplication rate, 32 plant growth regulator regimes were screened amongst which benzylaminopurine at 5.0 mg/l was found most suitable, yielding 100 % response and maximum number of shoots per explant (7.93); same concentration was also most supportive for repeated multiplication (6.53 shoots). The quality of the shoots and multiplication rate could be significantly enhanced (24.35 shoots) when adenine sulphate, glutamine and phloroglucinol, in an optimised concentration, were additionally supplemented. The clonal nature of the micropropagated plants was confirmed by random amplified polymorphic DNA analysis. A comparative analysis of AChEI activity was carried out amongst the methanolic extracts of stem segments of the mother plant, randomly selected seedlings of different age (4 and 6 months old) of the same mother plant and randomly selected micropropagated plants of different age (3 and 6 months age). The mother plant sample showed effective AChEI activity, with IC 50 of 66.46 μg/ml while seedlings, of different age groups, performed poorly (6-month-old seedlings, Se-1 6M , yielded IC 50 of 20,538.46 μg/ml, while two randomly selected 4 months' aged seedlings, Se-2 4M and Se-3 4M exhibited IC 50 of 19,341.03 and 24,281.70 μg/ml). On the other hand

  12. Outcome of total knee replacement following explantation and cemented spacer therapy

    Directory of Open Access Journals (Sweden)

    Ghanem, Mohamed

    2016-03-01

    Full Text Available Background: Infection after total knee replacement (TKR is one of the serious complications which must be pursued with a very effective therapeutic concept. In most cases this means revision arthroplasty, in which one-setting and two-setting procedures are distinguished. Healing of infection is the conditio sine qua non for re-implantation. This retrospective work presents an assessment of the success rate after a two-setting revision arthroplasty of the knee following periprosthetic infection. It further considers drawing conclusions concerning the optimal timing of re-implantation.Patients and methods: A total of 34 patients have been enclosed in this study from September 2005 to December 2013. 35 re-implantations were carried out following explantation of total knee and implantation of cemented spacer. The patient’s group comprised of 53% (18 males and 47% (16 females. The average age at re-implantation time was 72.2 years (ranging from 54 to 85 years. We particularly evaluated the microbial spectrum, the interval between explantation and re-implantation, the number of surgeries that were necessary prior to re-implantation as well as the postoperative course. Results: We reported 31.4% (11 reinfections following re-implantation surgeries. The number of the reinfections declined with increasing time interval between explantation and re-implantation. Patients who developed reinfections were operated on (re-implantation after an average of 4.47 months. Those patients with uncomplicated course were operated on (re-implantation after an average of 6.79 months. Nevertheless, we noticed no essential differences in outcome with regard to the number of surgeries carried out prior to re-implantation. Mobile spacers proved better outcome than temporary arthrodesis with intramedullary fixation.Conclusion: No uniform strategy of treatment exists after peri-prosthetic infections. In particular, no optimal timing can be stated concerning re

  13. In Vitro Multiplication of Stevia rebaudiana (Bertoni) Genotypes Using Different Explants

    OpenAIRE

    Seyis, Fatih; Yurteri, Emine; Özcan, Aysel

    2017-01-01

    In Vitro Multiplicationof Stevia rebaudiana (Bertoni)Genotypes by Using Different Explants Fatih Seyis1, EmineYurteri1, Aysel Özcan1 1Recep Tayyip Erdogan University, Faculty of Agronomyand Natural Sciences, Field Crops Department  Stevia rebaudiana Bertoni that is a member of the Compositae family is one of the most valuable  tropical medicinal plants. The origin of Stevia is South America, where it can be seen as a wild plant. Further it could be found in semi-arid habitat ranging from...

  14. Outcome of total knee replacement following explantation and cemented spacer therapy.

    Science.gov (United States)

    Ghanem, Mohamed; Zajonz, Dirk; Bollmann, Juliane; Geissler, Vanessa; Prietzel, Torsten; Moche, Michael; Roth, Andreas; Heyde, Christoph-E; Josten, Christoph

    2016-01-01

    Infection after total knee replacement (TKR) is one of the serious complications which must be pursued with a very effective therapeutic concept. In most cases this means revision arthroplasty, in which one-setting and two-setting procedures are distinguished. Healing of infection is the conditio sine qua non for re-implantation. This retrospective work presents an assessment of the success rate after a two-setting revision arthroplasty of the knee following periprosthetic infection. It further considers drawing conclusions concerning the optimal timing of re-implantation. A total of 34 patients have been enclosed in this study from September 2005 to December 2013. 35 re-implantations were carried out following explantation of total knee and implantation of cemented spacer. The patient's group comprised of 53% (18) males and 47% (16) females. The average age at re-implantation time was 72.2 years (ranging from 54 to 85 years). We particularly evaluated the microbial spectrum, the interval between explantation and re-implantation, the number of surgeries that were necessary prior to re-implantation as well as the postoperative course. We reported 31.4% (11) reinfections following re-implantation surgeries. The number of the reinfections declined with increasing time interval between explantation and re-implantation. Patients who developed reinfections were operated on (re-implantation) after an average of 4.47 months. Those patients with uncomplicated course were operated on (re-implantation) after an average of 6.79 months. Nevertheless, we noticed no essential differences in outcome with regard to the number of surgeries carried out prior to re-implantation. Mobile spacers proved better outcome than temporary arthrodesis with intramedullary fixation. No uniform strategy of treatment exists after peri-prosthetic infections. In particular, no optimal timing can be stated concerning re-implantation. Our data point out to the fact that a longer time interval between

  15. Stable genetic transformation of Jatropha curcas via Agrobacterium tumefaciens-mediated gene transfer using leaf explants

    KAUST Repository

    Kumar, Nitish; Vijay Anand, K.G.; Pamidimarri, D.V.N. Sudheer; Sarkar, Tanmoy; Reddy, Muppala P.; Radhakrishnan, T.; Kaul, Tanushri; Reddy, M.K.; Sopori, Sudhir K.

    2010-01-01

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel crop. A simple and reproducible protocol was developed for Agrobacterium tumefaciens-mediated stable genetic transformation of J. curcas using leaf explains. Agrobacterium strain LBA 4404 harbouring the binary vector pCAMBIA 1304 having sense-dehydration responsive element binding (S-DREB2A), beta-glucuronidase (gus), and hygromycin-phosphotransferase (hpt) genes were used for gene transfer. A number of parameters such as preculture of explains, wounding of leaf explants, Agrobacterium growth phase (OD), infection duration, co-cultivation period, co-cultivation medium pH, and acetosyringone, were studied to optimized transformation efficiency. The highest transformation efficiency was achieved using 4-day precultured, non-wounded leaf explants infected with Agrobacterium culture corresponding to OD(600)=0.6 for 20 min, followed by co-cultivation for 4 days in a co-cultivation medium containing 100 mu M acetosyringone, pH 5.7. Co-cultivated leaf explants were initially cultured on Murashige and Skoog (MS) medium supplemented with 2.27 mu M thidiazuron (TDZ) for regeneration of shoot buds, followed by selection on same medium with 5 mu g ml(-1) hygromycin. Selected shoot buds were transferred to MS medium containing 10 mu M kinetin (Kn), 4.5 mu M 6-benzyl aminopurine (BA), and 5.5 mu M alpha-naphthaleneacetic acid (NAA) for proliferation. The proliferated shoots were elongated on MS medium supplemented with 2.25 mu M BA and 8.5 mu M indole-3-acetic acid (IAA). The elongated shoots were rooted on half strength MS medium supplemented with 15 mu M indole-3-butyric acid (IBA), 5.7 mu M IAA, 5.5 mu M NAA, and 0.25 mg l(-1) activated charcoal. GUS histochemical analysis of the transgenic tissues further confirmed the transformation event. PCR and DNA gel blot hybridization were performed to confirm the presence of transgene. A transformation efficiency of 29% was

  16. Boron neutron capture therapy for an explanted organ: The logistical challenges

    International Nuclear Information System (INIS)

    Wittig, A.; Moss, R.; Kaiser, G.M.; Malago, M.; Nievaart, V.; Sauerwein, W.A.

    2009-01-01

    Single liver metastases of colorectal cancer can be cured by surgery; disseminated liver metastases are incurable. A research group in Pavia, Italy, used BNCT as an experimental method to irradiate in curative intention the explanted liver of patients suffering from disseminated hepatic metastases. The situation in Pavia, where a reactor with a specially adapted thermal column and the hospital are close by, is unique. For the purpose of the present study, it was necessary to investigate how the Pavia experience can be repeated with transplantation centers located at distance from a reactor. Some basic investigations of the logistics of such a procedure are reported.

  17. Stable genetic transformation of Jatropha curcas via Agrobacterium tumefaciens-mediated gene transfer using leaf explants

    KAUST Repository

    Kumar, Nitish

    2010-07-01

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel crop. A simple and reproducible protocol was developed for Agrobacterium tumefaciens-mediated stable genetic transformation of J. curcas using leaf explains. Agrobacterium strain LBA 4404 harbouring the binary vector pCAMBIA 1304 having sense-dehydration responsive element binding (S-DREB2A), beta-glucuronidase (gus), and hygromycin-phosphotransferase (hpt) genes were used for gene transfer. A number of parameters such as preculture of explains, wounding of leaf explants, Agrobacterium growth phase (OD), infection duration, co-cultivation period, co-cultivation medium pH, and acetosyringone, were studied to optimized transformation efficiency. The highest transformation efficiency was achieved using 4-day precultured, non-wounded leaf explants infected with Agrobacterium culture corresponding to OD(600)=0.6 for 20 min, followed by co-cultivation for 4 days in a co-cultivation medium containing 100 mu M acetosyringone, pH 5.7. Co-cultivated leaf explants were initially cultured on Murashige and Skoog (MS) medium supplemented with 2.27 mu M thidiazuron (TDZ) for regeneration of shoot buds, followed by selection on same medium with 5 mu g ml(-1) hygromycin. Selected shoot buds were transferred to MS medium containing 10 mu M kinetin (Kn), 4.5 mu M 6-benzyl aminopurine (BA), and 5.5 mu M alpha-naphthaleneacetic acid (NAA) for proliferation. The proliferated shoots were elongated on MS medium supplemented with 2.25 mu M BA and 8.5 mu M indole-3-acetic acid (IAA). The elongated shoots were rooted on half strength MS medium supplemented with 15 mu M indole-3-butyric acid (IBA), 5.7 mu M IAA, 5.5 mu M NAA, and 0.25 mg l(-1) activated charcoal. GUS histochemical analysis of the transgenic tissues further confirmed the transformation event. PCR and DNA gel blot hybridization were performed to confirm the presence of transgene. A transformation efficiency of 29% was

  18. High frequency plant regeneration from leaf explants derived callus of evening primrose (oenothera biennis)

    International Nuclear Information System (INIS)

    Ghauri, E.G.; Shafi, N.; Ghani, S.; Fatima, A.

    2008-01-01

    The seeds of Evening primrose were aseptically grown and leaf explants were used for establishment of callus culture. The Excellent growth in callus biomass was achieved on MS medium supplemented with 2, 4, -D and TDZ. For optimal growth of bud and shoot regeneration, fortification of IAA along with TDZ, or BAP was found to be essential. Rooting (70%) could be inducted on hormone free MS-medium. This percentage improved to 98 when NAA was added to the medium. The plantlets thus obtained were transferred to the field successfully after passing through the process of hardening. (author)

  19. Hypocotyl shape in radishes - a useful impact criterion for biological indication of ozone?; Hypokotyl-Form bei Radieschen - ein sinnvolles Wirkungskriterium fuer die Bioindikation von Ozon?

    Energy Technology Data Exchange (ETDEWEB)

    Kostka-Rick, R.

    1992-12-31

    A consistent correlation between certain `source` leaves and specific `sink` regions in the root of Beta vulgaris L. justifies to study the impact of ozone (O{sub 3}) on the shape of the hypocotyl in radish (Raphanus sativus L.) and the potential use of shape variants as effect criteria. A 7-day period of exposure to O{sub 3} ranging within realistic immission levels caused a nonsignificant reduction of hypocotyl fresh weight in radish. Two out three of the shape indices under study were also changed by ozone exposure - sometimes significantly. A discriminance function derived from several shape indices with or without fresh weight allowed a significant separation between the two O{sub 3}-treatment variants. Treatment with the anti-oxidant ethylene diurea (EDU) had no essential effect on hypocotyl shape. The author discusses the use of shape variants for ozone bio-indication. (orig.) [Deutsch] Eine konsistente Beziehung zwischen bestimmten `source`-Blaettern und spezifischen `sink`-Regionen in der Wurzel von Beta vulgaris L. rechtfertigt die Ueberpruefung des Einflusses von Ozon (O{sub 3}) auf die Form des Hypokotyls bei Radies (Raphanus sativus L.) und der potentiellen Nutzung von Formvariablen als Wirkungskriterium. Eine 7taegige O{sub 3}-Belastung im Bereich realistischer Immissionskonzentrationen verursachte eine nicht-signifikante Minderung des Hypokotyl-Frischgewichtes von Radies. Zwei von drei der untersuchten Formindizes wurden durch die Ozon-Belastung ebenfalls, z.T. signifikant, veraendert. Eine Diskriminanzfunktion aus mehreren Formindizes, mit oder ohne Einbeziehung des Frischgewichtes, gestattete eine signifikante Trennung zwischen den beiden O{sub 3}-Behandlungsvarianten. Eine Behandlung mit dem Anti-oxidants Ethylendiurea (EDU) hatte keinen wesentlichen Einfluss auf die Hypokotylform. Die Moeglichkeiten des Einsatzes von Formvariablen in der Bioindikation von Ozon werden diskutiert. (orig.)

  20. Organotypic Culture of Breast Tumor Explants as a Multicellular System for the Screening of Natural Compounds with Antineoplastic Potential

    Directory of Open Access Journals (Sweden)

    Irma Edith Carranza-Torres

    2015-01-01

    Full Text Available Breast cancer is the leading cause of death in women worldwide. The search for novel compounds with antitumor activity, with less adverse effects and higher efficacy, and the development of methods to evaluate their toxicity is an area of ​​intense research. In this study we implemented the preparation and culture of breast tumor explants, which were obtained from precision-cut breast tumor slices. In order to validate the model we are proposing to screen antineoplastic effect of natural compounds, we selected caffeic acid, ursolic acid, and rosmarinic acid. Using the Krumdieck tissue slicer, precision-cut tissue slices were prepared from breast cancer samples; from these slices, 4 mm explants were obtained and incubated with the selected compounds. Viability was assessed by Alamar Blue assay, LDH release, and histopathological criteria. Results showed that the viability of the explants cultured in the presence of paclitaxel (positive control decreased significantly (P<0.05; however, tumor samples responded differently to each compound. When the explants were coincubated with paclitaxel and compounds, a synergic effect was observed. This study shows that ex vivo culture of breast cancer explants offers a suitable alternative model for evaluating natural or synthetic compounds with antitumor properties within the complex microenvironment of the tumor.

  1. Adult Mouse DRG Explant and Dissociated Cell Models to Investigate Neuroplasticity and Responses to Environmental Insults Including Viral Infection.

    Science.gov (United States)

    Fornaro, Michele; Sharthiya, Harsh; Tiwari, Vaibhav

    2018-03-09

    This protocol describes an ex vivo model of mouse-derived dorsal root ganglia (DRG) explant and in vitro DRG-derived co-culture of dissociated sensory neurons and glial satellite cells. These are useful and versatile models to investigate a variety of biological responses associated with physiological and pathological conditions of the peripheral nervous system (PNS) ranging from neuron-glial interaction, neuroplasticity, neuroinflammation, and viral infection. The usage of DRG explant is scientifically advantageous compared to simplistic single cells models for multiple reasons. For instance, as an organotypic culture, the DRG explant allows ex vivo transfer of an entire neuronal network including the extracellular microenvironment that play a significant role in all the neuronal and glial functions. Further, DRG explants can also be maintained ex vivo for several days and the culture conditions can be perturbed as desired. In addition, the harvested DRG can be further dissociated into an in vitro co-culture of primary sensory neurons and satellite glial cells to investigate neuronal-glial interaction, neuritogenesis, axonal cone interaction with the extracellular microenvironment, and more general, any aspect associated with the neuronal metabolism. Therefore, the DRG-explant system offers a great deal of flexibility to study a wide array of events related to biological, physiological, and pathological conditions in a cost-effective manner.

  2. Micropropagation of Calophyllum brasiliense (Cambess.) from nodal segments.

    Science.gov (United States)

    Silveira, S S; Cordeiro-Silva, R; Degenhardt-Goldbach, J; Quoirin, M

    2016-05-03

    Micropropagation of Calophyllum brasiliense Cambess. (Clusiaceae) is a way to overcome difficulties in achieving large-scale plant production, given the recalcitrant nature of the seeds, irregular fructification and absence of natural vegetative propagation of the species. Cultures were established using nodal segments 2 cm in length, obtained from 1-2 year old seedlings, maintained in a greenhouse. Mercury chloride and Plant Preservative Mixture™ were used in the surface sterilizing stage, better results being achieved with Plant Preservative Mixture™ incorporation in culture medium, at any concentration. Polyvinylpyrrolidone, activated charcoal, cysteine, ascorbic acid or citric acid were added to the culture medium to avoid oxidation. After 30 days of culture, polyvinylpirrolidone and ascorbic acid gave better results, eliminating oxidation in most explants. For shoot multiplication, benzylaminopurine was used in concentrations of 4.4 and 8.8 µM in Woody Plant Medium, resulting in an average of 4.43 and 4.68 shoots per explant, respectively, after 90 days. Indole-3-butyric acid and α-naphthalene acetic acid were used to induce root formation, reaching a maximum rooting rate of 24% with 20µM α-naphthalene acetic acid. For acclimatization. the rooted plants were transferred to Plantmax® substrate and cultured in a greenhouse, reaching 79% of survival after 30 days and 60% after one year.

  3. All-trans retinoic acid inhibits craniopharyngioma cell growth: study on an explant cell model.

    Science.gov (United States)

    Li, Qiang; You, Chao; Zhou, Liangxue; Sima, Xiutian; Liu, Zhiyong; Liu, Hao; Xu, Jianguo

    2013-05-01

    The ratio between FABP5 and CRABPII determines cellular response to physiological level of retinoic acid; tumor cells undergo proliferation with high level of FABP5 and apoptosis with high level of CRABPII. We intended to study FABP5 and CRABPII expression in craniopharyngiomas, to establish craniopharyngioma cell model using explants method, and to study the effect of pharmacological dose of retinoic acid on craniopharyngioma cells. Expression of FABP5 and CRABPII in craniopharyngioma tissue from 20 patients was studied using immunohistochemistry. Primary craniopharyngioma cell cultures were established using tissue explants method. Craniopharyngioma cells were treated using various concentrations of all-trans retinoic acid, and cell growth curve, apoptosis, expression of FABP5, CRABPII and NF-κB were assayed in different groups. FABP5/CRABPII ratio was significantly higher in adamatinomatous group than that in papillary group. Cell cultures were established in 19 cases (95 %). Pharmacological level retinoic acid inhibited cell growth and induced cellular apoptosis in dose dependent manner, and apoptosis rate cells treated with 30 μM retinoic acid for 24 h was 43 %. Also, retinoic acid increased CRABPII, and decreased FABP5 and NF-κB expression in craniopharyngioma cells. High FABP5/CRABPII ratio is observed in adamatinomatous craniopharyngioma. Retinoic acid at pharmacological level induced craniopharyngioma cell apoptosis via increasing FABP5/CRABPII ratio and inhibiting NF-κB signaling pathway. Our study demonstrated that all-trans retinoic acid might be a candidate for craniopharyngioma adjuvant chemotherapy in future.

  4. An efficient regeneration and rapid micropropagation protocol for Almond using dormant axillary buds as explants.

    Science.gov (United States)

    Choudhary, Ravish; Chaudhury, Rekha; Malik, Surendra Kumar; Sharma, Kailash Chandra

    2015-07-01

    An efficient in vitro protocol was standardized for Almond (Prunus dulcis) propagation using dormant axillary buds as explants. Explants were cultured on Murashige and Skoog (MS) and woody plant medium (WPM) supplemented with different concentration/combination(s) of phytohormones. MS basal medium showed lowest shoot induction and took longest duration for shoot initiation. Multiple shoots were induced in MS medium supplemented with the combination of BAP (0.5 mgL(-1)). Cultures showed poor response for rooting in all combinations of plant growth regulators (PGRs) and took 90 days for initiation. Rooting was higher in half strength of MS than in full-strength. The highest root induction (33.33%) was recorded in half MS medium supplemented with 0.1 mgL(-1) IBA (indole-3-butyric acid) followed by full strength of MS medium (20%) supplemented with IBA (0.1 mgL(-1)). α-Naphthalene acetic acid (NAA) was less effective for rooting than IBA. The highest root induction (25%) was found in half strength of MS medium supplemented with 0.1 mgL(-1) NAA followed by full strength of MS medium (20%). The protocol developed would be of use in mass propagation of almond and also support in vitro conservation.

  5. Effect of explant density and medium culture volumes on cassava micropropagation in Temporal Immersion System

    Directory of Open Access Journals (Sweden)

    Milagros Basail

    2003-04-01

    Full Text Available Due to the need of producing high quality planting material available to cassava growers, it has been necessary to look for alternatives in order to increase the efficiancy of in vitro propagation methods and their automation, such as the use of the Temporal Immersion Systems (RITA®. This work was carried out to increase the multiplication coefficient for cassava mass propagation through out Temporal Immersion Systems. The clone ‘CMC-40’ was used. Different medium volumes per explant, and material density per unit at a given Immersion frequency were tested. The highest results were obtained in the 2.8 multiplication coefficient with 20 ml culture medium volume and 3.2 using a density of 40 explants/flask. When the Temporal Immersion System is used with these results, a more efficient method for cassava micropropagation is established and also higher quality vitroplants for the rooting stage and further acclimatization in field conditions are produced. Key Words: Tissue Culture, liquid culture medium, Manihot esculenta Crantz

  6. Histological and molecular evaluation of patient-derived colorectal cancer explants.

    Directory of Open Access Journals (Sweden)

    Joshua M Uronis

    Full Text Available Mouse models have been developed to investigate colorectal cancer etiology and evaluate new anti-cancer therapies. While genetically engineered and carcinogen-induced mouse models have provided important information with regard to the mechanisms underlying the oncogenic process, tumor xenograft models remain the standard for the evaluation of new chemotherapy and targeted drug treatments for clinical use. However, it remains unclear to what extent explanted colorectal tumor tissues retain inherent pathological features over time. In this study, we have generated a panel of 27 patient-derived colorectal cancer explants (PDCCEs by direct transplantation of human colorectal cancer tissues into NOD-SCID mice. Using this panel, we performed a comparison of histology, gene expression and mutation status between PDCCEs and the original human tissues from which they were derived. Our findings demonstrate that PDCCEs maintain key histological features, basic gene expression patterns and KRAS/BRAF mutation status through multiple passages. Altogether, these findings suggest that PDCCEs maintain similarity to the patient tumor from which they are derived and may have the potential to serve as a reliable preclinical model that can be incorporated into future strategies to optimize individual therapy for patients with colorectal cancer.

  7. Generation and characterisation of human umbilical cord derived mesenchymal stem cells by explant method.

    Science.gov (United States)

    Yusoff, Z; Maqbool, M; George, E; Hassan, R; Ramasamy, R

    2016-06-01

    Mesenchymal stem cells (MSCs) derived from human umbilical cord (UC) have been considered as an important tool for treating various malignancies, tissue repair and organ regeneration. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) are better alternative to MSCs that derived from bone marrow (BM-MSCs) as they are regarded as medical waste with little ethical concern for research and easily culture-expanded. In this present study, the foetal distal end of human UC was utilised to generate MSC by explant method. Upon in vitro culture, adherent cells with fibroblastic morphology were generated with rapid growth kinetics. Under the respective inductive conditions, these cells were capable of differentiating into adipocytes and osteocytes; express an array of standard MSC's surface markers CD29, CD73, CD90, CD106 and MHC-class I. Further assessment of immunosuppression activity revealed that MSCs generated from UC had profoundly inhibited the proliferation of mitogen-activated T lymphocytes in a dosedependent manner. The current laboratory findings have reinforced the application of explant method to generate UCMSCs thus, exploring an ideal platform to fulfil the increasing demand of MSCs for research and potential clinical use.

  8. Automated Image Analysis of Lung Branching Morphogenesis from Microscopic Images of Fetal Rat Explants

    Science.gov (United States)

    Rodrigues, Pedro L.; Rodrigues, Nuno F.; Duque, Duarte; Granja, Sara; Correia-Pinto, Jorge; Vilaça, João L.

    2014-01-01

    Background. Regulating mechanisms of branching morphogenesis of fetal lung rat explants have been an essential tool for molecular research. This work presents a new methodology to accurately quantify the epithelial, outer contour, and peripheral airway buds of lung explants during cellular development from microscopic images. Methods. The outer contour was defined using an adaptive and multiscale threshold algorithm whose level was automatically calculated based on an entropy maximization criterion. The inner lung epithelium was defined by a clustering procedure that groups small image regions according to the minimum description length principle and local statistical properties. Finally, the number of peripheral buds was counted as the skeleton branched ends from a skeletonized image of the lung inner epithelia. Results. The time for lung branching morphometric analysis was reduced in 98% in contrast to the manual method. Best results were obtained in the first two days of cellular development, with lesser standard deviations. Nonsignificant differences were found between the automatic and manual results in all culture days. Conclusions. The proposed method introduces a series of advantages related to its intuitive use and accuracy, making the technique suitable to images with different lighting characteristics and allowing a reliable comparison between different researchers. PMID:25250057

  9. Automated Image Analysis of Lung Branching Morphogenesis from Microscopic Images of Fetal Rat Explants

    Directory of Open Access Journals (Sweden)

    Pedro L. Rodrigues

    2014-01-01

    Full Text Available Background. Regulating mechanisms of branching morphogenesis of fetal lung rat explants have been an essential tool for molecular research. This work presents a new methodology to accurately quantify the epithelial, outer contour, and peripheral airway buds of lung explants during cellular development from microscopic images. Methods. The outer contour was defined using an adaptive and multiscale threshold algorithm whose level was automatically calculated based on an entropy maximization criterion. The inner lung epithelium was defined by a clustering procedure that groups small image regions according to the minimum description length principle and local statistical properties. Finally, the number of peripheral buds was counted as the skeleton branched ends from a skeletonized image of the lung inner epithelia. Results. The time for lung branching morphometric analysis was reduced in 98% in contrast to the manual method. Best results were obtained in the first two days of cellular development, with lesser standard deviations. Nonsignificant differences were found between the automatic and manual results in all culture days. Conclusions. The proposed method introduces a series of advantages related to its intuitive use and accuracy, making the technique suitable to images with different lighting characteristics and allowing a reliable comparison between different researchers.

  10. Establishing axenic cultures from mature pecan embryo explants on media with low water availability.

    Science.gov (United States)

    Obeidy, A A; Smith, M A

    1990-12-01

    Endophytic fungi associated with mature pecan (Carya illinoensis (Wangenh.) C. Koch) nuts prevented successful, contaminant-free in vitro culture of embryo expiants, even after rigorous surface disinfestation of the nuts and careful aseptic shelling. Disinfestation with sodium hypochlorite after shell removal was also unsuccessful, because even dilute concentrations which were ineffective against the fungal contaminants prevented subsequent growth from the embryo. Explanting media with low water availability which would not sustain growth of fungal contaminants, but supported growth from mature pecan embryos, were developed as an alternative disinfestation method. The explanting media were supplemented with 0.9-1.5% agar, and other media components were selectively omitted to test their influence on water availability and fungal growth. Disinfestation of up to 65% of the cultures was accomplished, depending on the medium formulation, compared to 100% loss to contamination on control medium (0.5% agar). A complete medium (containing sucrose, salts, vitamins, 18 μM BAP, and 5 μM IBA) with 1.5% agar provided control of contamination, and encouraged subsequent regeneration from the embryo expiants, which remained free of contaminant growth through subsequent subcultures.

  11. Bystander-induced apoptosis and premature differentiation in primary urothelial explants after charged particle microbeam irradiation

    International Nuclear Information System (INIS)

    Belyakov, O.V.; Folkard, M.; Mothersill, C.; Prise, K.M.; Michael, B.D.

    2002-01-01

    The ureter primary explant technique was developed to study bystander effects under in vivo like conditions where stem and differentiated cells are present. Irradiation was performed with a 3 He 2+ charged particle microbeam available at the Gray Cancer Institute, with high (∼2 μm) precision. Tissue sections from porcine ureters were pre-irradiated with the microbeam at a single location with 10 3 He 2+ particles (5 MeV; LET 70 keV.μm -1 ). After irradiation, the tissue section was incubated for 7 days, thus allowing the explant outgrowth to form. Total cellular damage (total fraction of micronucleated and apoptotic cells) was measured according to morphological criteria. Apoptosis was also assessed using a 3'-OH DNA end-labelling technique. Premature differentiation was estimated using antibodies to uroplakin III, a specific marker of terminal urothelial differentiation. Results of our experiments demonstrated a significant bystander-induced differentiation and a less significant increase in apoptotic and micronucleated cells. A hypothesis based on the protective nature of the bystander effect is proposed. (author)

  12. In vitro plant regeneration from leaf explants of Solanum pimpinellifolium L.

    Directory of Open Access Journals (Sweden)

    Shirley Valderrama-Alfaro

    2011-01-01

    Full Text Available Tomato is a species of agricultural importance. Besides, it is a source of vitamins, minerals and antioxidant compounds. Therefore it is essential to obtain varieties resistant to diseases. Solanum pimpinellifolium L. ‘Tomatillo wild’, one of the wild relatives of tomato, is considered multipurpose reservoir of genes. This characteristic must be exploited. In vitro plantlets from leaves of ‘wild tomatillo’ were regenerated with this objective. A system for in vitro germination of seeds obtained from ex vitro sources was created. The use of seed allowed obtaining adequate seedlings in a short time to start regeneration. Thein vitro response of explants was evaluated in four treatments. The Murashige and Skoog (MS basal culture medium supplemented with a-naphthaleneacetic acid (NAA and 6-benzyl aminopurine (BAP was used in different combinations. The higher percentage (30% of shoot induction was achieved with 0.1 mg l1 NAA / 1 mg l1 BAP. The presence of callus and roots was observed after seven days of culture in the combination treatment with 1 mg l1 NAA / 0.1 mg l1 BAP. Callus showed 1 or 2 adventitious shoots per explant after 30 days of culture and 3 to 6 shoots after seventy days. Furthermore, the presence of fully formed adventitious plantlets (shoot and root was observed after the fifth week of culture. Therefore, the best combination for in vitro regeneration is NAA 0.1 mg l1 / 1 mg l1 BAP. Keywords: auxin, cytokinins, indirect organogenesis

  13. Chitinase, beta-1,3-glucanase, osmotin, and extensin are expressed in tobacco explants during flower formation

    DEFF Research Database (Denmark)

    Neale, A D; Wahleithner, J A; Lund, Marianne

    1990-01-01

    be considered a pathogenesis-related protein. These genes, which were highly expressed in explants during de novo flower formation but not in explants forming vegetative shoots [Meeks-Wagner et al. (1989). Plant Cell 1, 25-35], were also regulated developmentally in day-neutral and photoresponsive tobacco......Sequence analysis of five gene families that were isolated from tobacco thin cell layer explants initiating floral development [Meeks-Wagner et al. (1989). Plant Cell 1, 25-35] showed that two encode the pathogenesis-related proteins basic chitinase and basic beta-1,3-glucanase, while a third...... encodes the cell wall protein extensin, which also accumulates during pathogen attack. Another sequence family encodes the water stress-induced protein osmotin [Singh et al. (1989). Plant Physiol. 90, 1096-1101]. We found that osmotin was also induced by viral infection and wounding and, hence, could...

  14. [Explantation method of isolating a persistent tick-borne encephalitis virus from the organs of infected monkeys].

    Science.gov (United States)

    Levina, L S; Pogodina, V V

    1981-01-01

    The method of explantation was used to examine 63 organs from M. rhesus monkeys 92-783 days after intracerebral and subcutaneous inoculation with the Vasilchenko, Aina/1448 and 41/65 strains of tick-borne encephalitis virus. The optimal time for examination of the explants by tests of the hemagglutinating, cytopathogenic activity of the virus and its pathogenicity for mice was found to be the 15th day of cultivation. A comparative study of the properties of 3 isolates obtained from explants of the spleen, liver and subcortical cerebral ganglia 202 and 307 days after inoculation of monkeys was carried out. The isolates differed from the parental TBE virus strains by their capacity to form small plaques in PEKV cell cultures (pig embryo kidney cells in versen medium).

  15. Selection of valine-resistance in callus culture of Arabidopsis thaliana (L. Heynh. derived from leaf explants

    Directory of Open Access Journals (Sweden)

    Małgorzata D. Gaj

    2014-01-01

    Full Text Available The selection of valine-resistant mutants was carried out in leaf explant cultures of three Arabidopsis thaliana (L. Heynh. ecotypes: C-24, RLD and Columbia. The valine concentration used for in vitro selection, lethal for seed-growing plants, has not affected callus formation and growth. However, strong inhibition of shoot regeneration ability of calli growing under selection pressure was noticed. In total, 1043 explants were cultured on valine medium and 18 shoots were regenerated with an average frequency of 1.7 shoots per 100 calli. Most R1 shoots were sterile and seeds were collected from 3 plants. The transmission of valine-resistance to the sexual progeny of these plants was scored and the increased level of valine-resistance was found in progeny of one line - 61 C. This line originated from the culture of Columbia leaf explant and displayed tetraploid chromosome number.

  16. Evaluation of cell wall preparations for proteomics: a new procedure for purifying cell walls from Arabidopsis hypocotyls

    Directory of Open Access Journals (Sweden)

    Canut Hervé

    2006-05-01

    Full Text Available Abstract Background The ultimate goal of proteomic analysis of a cell compartment should be the exhaustive identification of resident proteins; excluding proteins from other cell compartments. Reaching such a goal closely depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific difficulties: (i the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP during the isolation procedure, (ii polysaccharide networks of cellulose, hemicelluloses and pectins form potential traps for contaminants such as intracellular proteins. Several reported procedures to isolate cell walls for proteomic analyses led to the isolation of a high proportion (more than 50% of predicted intracellular proteins. Since isolated cell walls should hold secreted proteins, one can imagine alternative procedures to prepare cell walls containing a lower proportion of contaminant proteins. Results The rationales of several published procedures to isolate cell walls for proteomics were analyzed, with regard to the bioinformatic-predicted subcellular localization of the identified proteins. Critical steps were revealed: (i homogenization in low ionic strength acid buffer to retain CWP, (ii purification through increasing density cushions, (iii extensive washes with a low ionic strength acid buffer to retain CWP while removing as many cytosolic proteins as possible, and (iv absence of detergents. A new procedure was developed to prepare cell walls from etiolated hypocotyls of Arabidopsis thaliana. After salt extraction, a high proportion of proteins predicted to be secreted was released (73%, belonging to the same functional classes as proteins identified using previously described protocols. Finally, removal of intracellular proteins was obtained using detergents, but their amount represented less than 3% in mass of the total protein extract, based on protein quantification. Conclusion The

  17. Segmentation, advertising and prices

    NARCIS (Netherlands)

    Galeotti, Andrea; Moraga González, José

    This paper explores the implications of market segmentation on firm competitiveness. In contrast to earlier work, here market segmentation is minimal in the sense that it is based on consumer attributes that are completely unrelated to tastes. We show that when the market is comprised by two

  18. Sipunculans and segmentation

    DEFF Research Database (Denmark)

    Wanninger, Andreas; Kristof, Alen; Brinkmann, Nora

    2009-01-01

    mechanisms may act on the level of gene expression, cell proliferation, tissue differentiation and organ system formation in individual segments. Accordingly, in some polychaete annelids the first three pairs of segmental peripheral neurons arise synchronously, while the metameric commissures of the ventral...

  19. Enamel formation in vitro in mouse molar explants exposed to amelogenin polypeptides ATMP and LRAP on enamel development.

    Science.gov (United States)

    Ravindranath, Rajeswari M H; Devarajan, Asokan; Bringas, Pablo

    2007-12-01

    The enamel matrix contains amelogenin, leucine-rich amelogenin-polypeptide (LRAP), resulting from alternative splicing of the primary amelogenin-RNA transcript and tyrosine-rich amelogenin-polypeptide (TRAP), a proteolytic product of amelogenin. Presence of amelogenin-trityrosyl-motif peptide (ATMP) distinguishes TRAP from LRAP. The roles of these polypeptides in the formation of enamel remain to be elucidated. The mouse in vitro molar tooth-organ developed from bud stage (E16) was exposed to LRAP, ATMP, and mutated ATMP (T-ATMP, third proline replaced by threonine). The histology and morphometry of the explants on day-12 in culture was examined using Mallory's stain. Guanidine-HCl soluble protein concentrations of explants were compared. The enamel width and protein solubility indicate that the explant on day-12 is comparable to postnatal molar on day-3 in vivo. The enamel of both untreated explants as well as that in vivo is fuchinophilic (acid fuchsin, AF+). ATMP reduced the ameloblast-height, accumulated AF+ spherules at the apical end of ameloblasts, and disrupted enamel-dentin bonding. T-ATMP abrogated deposition of AF+ material on the aniline blue positive (AB+) enamel matrix. LRAP reduced ameloblast-height, increased the enamel-width without disruption (at 17.25 nmol) and increased the density of AF+ dentinal tubules. AF+ substance from the tubules is released onto the surface of the dentin. The Guanidine-HCl-soluble protein is elevated in ATMP-treated explants but decreased in LRAP-treated explants. Exogenous ATMP, T-ATMP and LRAP have divergent effects on developing enamel. Exogenous ATMP, but not LRAP, abrogates enamel-dentin bonding at 17.25 nmol. LRAP may play a role in the differentiation of ameloblasts, growth of enamel and formation of dentinal tubules.

  20. Agrobacterium-Mediated Transformation of Leaf Base Segments.

    Science.gov (United States)

    Gasparis, Sebastian

    2017-01-01

    Agrobacterium-mediated transformation has become a routine method of genetic engineering of cereals, gradually replacing the biolistic protocols. Simple integration patterns of transgenic loci, decent transformation efficiency, and technical simplicity are the main advantages offered by this method. Here we present a detailed protocol for the production of transgenic oat plants by Agrobacterium-mediated transformation of leaf base segments. The use of leaf explants as target tissues for transformation and in vitro regeneration of transgenic plants may be a good alternative for genotypes which are not susceptible to regeneration from immature or mature embryos. We also describe the biochemical and molecular analysis procedures of the transgenic plants including a GUS histochemical assay, and Southern blot, both of which are optimized for application in oat.

  1. Activation of anthocyanin synthesis genes by white light in eggplant hypocotyl tissues, and identification of an inducible P-450 cDNA

    International Nuclear Information System (INIS)

    Toguri, T.; Umemoto, N.; Kobayashi, O.; Ohtani, T.

    1993-01-01

    Eggplant seedlings (Solanum melongena) grown under red light irradiation showed a normal morphology with green, fully expanded cotyledons. When the seedlings grown under red light were irradiated with ultraviolet-containing white light, anthocyanin synthesis was induced in the hypocotyl tissues, especially when a UV light supplement was added. The accumulation of pigments was closely associated with the expression of genes involved in flavonoid synthesis. These genes include chalcone synthase (CHS) and dihydroflavonol 4-reductase (DFR). Using subtracted probes, which had been enriched for the accumulated mRNA, one white light-responsive cDNA was identified as being a P450 gene by comparison with database sequences. The maximal amino acid homology this cDNA had with other P450s was 36%. This was with CYP71 from avocado (Persea americana). Thus it represents a new P-450 family, which has been named CYP75. The mRNA of this gene was localized in the hypocotyl tissues of eggplant seedlings, which had been white light-irradiated. The transcript was accumulated by changing the light source, as in the case of other flavonoid biosynthesis genes. In delphinidin producing petunia plants, the mRNAs corresponding to the eggplant P-450 and flavonoid biosynthesis genes such as CHS and DFR were most abundant during the mid stage of flower bud development, but could not be detected in leaf tissues. These results suggest that this P-450 gene encodes a hydroxylating enzyme involved in flavonoid biosynthesis. (author)

  2. In vitro propagation of olive (Olea europaea L.) by nodal segmentation of elongated shoots.

    Science.gov (United States)

    Lambardi, Maurizio; Ozudogru, Elif Aylin; Roncasaglia, Romano

    2013-01-01

    Olive (Olea europaea L.), long-living, ever-green fruit tree of the Old World, has been part of a traditional landscape in the Mediterranean area for centuries. Both the fruits consumed after processing and the oil extracted from the fruits are among the main components of the Mediterranean diet, widely used for salads and cooking, as well as for preserving other food. Documentations show that the ancient use of this beautiful tree also includes lamp fuel production, wool treatment, soap production, medicine, and cosmetics. However, unlike the majority of the fruit species, olive propagation is still a laborious practice. As regards traditional propagation, rooting of cuttings and grafting stem segments onto rootstocks are possible, former being achieved only when the cuttings are collected in specific periods (spring or beginning of autumn), and latter only when skilled grafters are available. In both the cases, performance of the cultivars varies considerably. The regeneration of whole plants from ovules, on the other hand, is used only occasionally. Micropropagation of olive is not easy mainly due to explant oxidation, difficulties in explant disinfection, and labor-oriented establishment of in vitro shoot cultures. However today, the progress in micropropagation technology has made available the complete protocols for several Mediterranean cultivars. This chapter describes a micropropagation protocol based on the segmentation of nodal segments obtained from elongated shoots.

  3. Pancreas and cyst segmentation

    Science.gov (United States)

    Dmitriev, Konstantin; Gutenko, Ievgeniia; Nadeem, Saad; Kaufman, Arie

    2016-03-01

    Accurate segmentation of abdominal organs from medical images is an essential part of surgical planning and computer-aided disease diagnosis. Many existing algorithms are specialized for the segmentation of healthy organs. Cystic pancreas segmentation is especially challenging due to its low contrast boundaries, variability in shape, location and the stage of the pancreatic cancer. We present a semi-automatic segmentation algorithm for pancreata with cysts. In contrast to existing automatic segmentation approaches for healthy pancreas segmentation which are amenable to atlas/statistical shape approaches, a pancreas with cysts can have even higher variability with respect to the shape of the pancreas due to the size and shape of the cyst(s). Hence, fine results are better attained with semi-automatic steerable approaches. We use a novel combination of random walker and region growing approaches to delineate the boundaries of the pancreas and cysts with respective best Dice coefficients of 85.1% and 86.7%, and respective best volumetric overlap errors of 26.0% and 23.5%. Results show that the proposed algorithm for pancreas and pancreatic cyst segmentation is accurate and stable.

  4. Segmentation of consumer's markets and evaluation of market's segments

    OpenAIRE

    ŠVECOVÁ, Iveta

    2013-01-01

    The goal of this bachelor thesis was to explain a possibly segmentation of consumer´s markets for a chosen company, and to present a suitable goods offer, so it would be suitable to the needs of selected segments. The work is divided into theoretical and practical part. First part describes marketing, segmentation, segmentation of consumer's markets, consumer's market, market's segments a other terms. Second part describes an evaluation of questionnaire survey, discovering of market's segment...

  5. Micropropagation of Origanum acutidens (HAND.-MAZZ.) IETSWAART using stem node explants.

    Science.gov (United States)

    Yildirim, Mehmet Ugur

    2013-01-01

    Origanum acutidens (HAND.-MAZZ.) IETSWAART is a promising ornamental plant that can be widely used in landscape management. It is endemic to Eastern Anatolian region of Turkey. Tissue culture has not been used to micropropagate it. The study reports stem node explants from one-week-old seedlings of the plant for successful micropropagation. The stem nodes were cultured on MS medium containing 0.6, 1.2, 1.8, and 2.4 mg/L BAP with 0.2 mg/L NAA. Visible effects of culture media on shoot proliferation were recorded. Shoot regeneration rate was maximum on MS medium containing 1.80 mg/L BAP-0.2 mg/L NAA. The micropropagated shoots were rooted on MS medium containing 0.2 mg/L NAA. All microrooted plantlets survived during acclimatisation on peat moss. It was concluded that O. acutidens can be successfully micropropagated under in vitro conditions.

  6. Postoperative diffuse opacification of a hydrophilic acrylic intraocular lens: analysis of an explant.

    Science.gov (United States)

    Cavallini, Gian Maria; Volante, Veronica; Campi, Luca; De Maria, Michele; Fornasari, Elisa; Urso, Giancarlo

    2017-06-14

    We describe the clinicopathological and ultrastructural features of an opaque single-piece hydrophilic acrylic intraocular lens (IOL) explanted from a patient. The main outcome of this report is the documentation of calcium deposits confirmed by surface analysis. The decrease in visual acuity was due to the opacification of the IOL. The opacification involved both the optic plate and the haptics. The analysis at the scansion electron microscope revealed that the opacity was caused by the deposition of calcium and phosphate within the lens optic and haptics. This is the first case about the opacification of an Oculentis L-313. The opacification was characterized by calcium and phosphate deposition probably due to a morphological alteration of the posterior surface of the IOL.

  7. Somatic embryogenesis and plant regeneration in Carica papaya L. tissue culture derived from root explants.

    Science.gov (United States)

    Chen, M H; Wang, P J; Maeda, E

    1987-10-01

    The regeneration potential of shoot tip, stem, leaf, cotyledon and root explants of two papaya cultivars (Carica papaya cv. 'Solo' and cv. 'Sunrise') were studed. Callus induction of these two cultivars of papaya showed that the shoot tips and stems are most suitable for forming callus, while leaves, cotyledons and roots are comparatively difficult to induce callus. Callus induction also varied with the varities. Somatic embryogenesis was obtained from 3-month-old root cultures. A medium containing half strength of MS inorganic salts, 160 mg/l adenine sulfate, 1.0 mg/1 NAA, 0.5 mg/1 kinetin and 1.0 mg/1 GA3 was optimal for embryogenesis. The callus maintained high regenerative capacity after two years of culture on this medium. Plants derived from somatic embryos were obtained under green-house conditions.

  8. A novel 3D skin explant model to study anaerobic bacterial infection

    DEFF Research Database (Denmark)

    Maboni, Grazieli; Davenport, Rebecca; Sessford, Kate

    2017-01-01

    of the tissue structure and the cell types present in the host environment. A 3D skin culture model can be set up using tissues acquired from surgical procedures or post slaughter, making it a cost effective and attractive alternative to animal experimentation. The majority of 3D culture models have been......Skin infection studies are often limited by financial and ethical constraints, and alternatives, such as monolayer cell culture, do not reflect many cellular processes limiting their application. For a more functional replacement, 3D skin culture models offer many advantages such as the maintenance...... bacterium and the causative agent of footrot. The mechanism of infection and host immune response to D. nodosus is poorly understood. Here we present a novel 3D skin ex vivo model to study anaerobic bacterial infections using ovine skin explants infected with D. nodosus. Our results demonstrate that D...

  9. Micropropagation from cultured nodal explants of rose (Rosa hybrida L. cv. ‘Perfume Delight’

    Directory of Open Access Journals (Sweden)

    Kamnoon Kanchanapoom

    2010-01-01

    Full Text Available A method for the micropropagation of rose (Rosa hybrida L. cv. ‘Perfume Delight’ was developed. First to fifth nodal explants from young healthy shoots were excised and cultured on basal medium of Murashige and Skoog (1962, MS containing several concentrations of BA and NAA. Multiple shoot formation of up to 3 shoots was obtained on MS medium supplemented with 3 mg/l BA and 0.003 mg/l NAA. Shoot readily rooted on ¼MS medium devoid of growth regulators.Rooted plantlets were hardened and established in pots at 100% survival. In vitro flowering was observed on rose plantscultured on MS medium containing 3 mg/l BA and 0.003 mg/l NAA.

  10. Sprifermin (rhFGF18) modulates extracellular matrix turnover in cartilage explants ex vivo

    DEFF Research Database (Denmark)

    Reker, Ditte; Kjelgaard-Petersen, Cecilie Freja; Siebuhr, Anne Sofie

    2017-01-01

    Background: Sprifermin (recombinant human fibroblast growth factor 18) is in clinical development as a potential disease-modifying osteoarthritis drug (DMOAD). In vitro studies have shown that cartilage regenerative properties of sprifermin involve chondrocyte proliferation and extracellular matrix...... or placebo at weekly intervals, similar to the dosing regimen used in clinical trials. Pre-culturing with oncostatin M and tumour necrosis factor-a, was also used to induce an inflammatory state before treatment. Metabolic activity was measured using AlamarBlue, and chondrocyte proliferation was visualized...... aggrecanase activity. Results: Sprifermin was able to reach the chondrocytes through the extracellular matrix, as it increased cell proliferation and metabolic activity of explants. ProC2 and CS846 was dose-dependently increased (P

  11. Callus induction and plant regeneration from different explant types of Miscanthus x ogiformis Honda 'Giganteus'

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted; Petersen, Karen Koefoed

    1996-01-01

    . The explants were cultured on urashige and Skoog medium supplemented with 4.5, 13.6, 22.6 or 31.7 μM 2,4-dichlorophenoxyacetic acid. Three types of callus were formed but only one was embryogenic and regenerated plants. Callus induction and formation of embryogenic callus depended on the type and developmental......-propagated shoots and older leaves of greenhouse-grown plants. Immature inflorescences smaller than 2.5 cm produced a higher percentage of embryogenic callus than larger more mature inflorescences. Embryogenic callus derived from immature inflorescences had the highest regeneration capacity. Differences in 2......,4-dichlorophenoxyacetic acid concentrations had no significant effect on callus induction, embryogenic callus formation and plant regeneration....

  12. Callus induction and biomass accumulation in vitro in explants from chokeberry (Aronia melanocarpa (Michx. Elliot fruit

    Directory of Open Access Journals (Sweden)

    Tatiana I. Calalb

    2014-09-01

    Full Text Available In this study, the following features were determined: biological (the optimal histogen as explant and the optimal age of donor fruit, biotechnological (type, dosage and combination of growth regulators supplements in culture medium Murashige and Skoog as well as sucrose dosage, and physical (light regime, to induce callusing and biomass accumulation in vitro from the succulent chokeberry (Aronia melanocarpa (Michx. Elliot fruit. It turned out that it was much easier to induce callus from explants composed of the epicarp and hypoderm cut from fruits at 50–60 days after flowering. The role of light regime and varied supplementation of the basic MS medium with different doses of growth regulators was established; they resulted in four pigmented carpomass: violet, cream-pink, cream-white and green. The best combinations for the proliferation of fruit callus were culture media with 0.2–2.5 mg × dm-3 2,4-D+0.5 mg × dm-3 KIN +60 g × dm-3sucrose, while for fruit biomass accumulation enriched with phenolic substances – 2.5–3.5 mg × dm-3 NAA+0.5 mg × dm-3 KIN+60 g × dm-3sucrose. The chemical study of phenolic compounds by HPLC coupled with the mass spectrometry method identified chlorogenic acid, hiperozide, quercetrin, isoquercitrin and rutozide quantitatively and qualitatively in all pigmented carpomass and fruits; an exception is p-coumaric present only qualitatively in green carpomass and absent in fruit and quercetol absent in green carpomass.

  13. Effects of high hydrostatic pressure on bacterial growth on human ossicles explanted from cholesteatoma patients.

    Directory of Open Access Journals (Sweden)

    Steffen Dommerich

    Full Text Available BACKGROUND: High hydrostatic pressure (HHP treatment can eliminate cholesteatoma cells from explanted human ossicles prior to re-insertion. We analyzed the effects of HHP treatment on the microbial flora on ossicles and on the planktonic and biofilm states of selected isolates. METHODOLOGY: Twenty-six ossicles were explanted from cholesteatoma patients. Five ossicles were directly analyzed for microbial growth without further treatment. Fifteen ossicles were cut into two pieces. One piece was exposed to HHP of 350 MPa for 10 minutes. Both the treated and untreated (control pieces were then assessed semi-quantitatively. Three ossicles were cut into two pieces and exposed to identical pressure conditions with or without the addition of one of two different combinations of antibiotics to the medium. Differential effects of 10-minute in vitro exposure of planktonic and biofilm bacteria to pressures of 100 MPa, 250 MPa, 400 MPa and 540 MPa in isotonic and hypotonic media were analyzed using two patient isolates of Staphylococcus epidermidis and Neisseria subflava. Bacterial cell inactivation and biofilm destruction were assessed by colony counting and electron microscopy. PRINCIPAL FINDINGS: A variety of microorganisms were isolated from the ossicles. Irrespective of the medium, HHP treatment at 350 MPa for 10 minutes led to satisfying but incomplete inactivation especially of gram-negative bacteria. The addition of antibiotics increased the efficacy of elimination. A comparison of HHP treatment of planktonic and biofilm cells showed that the effects of HPP were reduced by about one decadic logarithmic unit when HPP was applied to biofilms. High hydrostatic pressure conditions that are suitable to inactivate cholesteatoma cells fail to completely sterilize ossicles even if antibiotics are added. As a result of the reduced microbial load and the viability loss of surviving bacteria, however, there is a lower risk of re-infection after re-insertion.

  14. Effects of High Hydrostatic Pressure on Bacterial Growth on Human Ossicles Explanted from Cholesteatoma Patients

    Science.gov (United States)

    Ostwald, Jürgen; Lindner, Tobias; Zautner, Andreas Erich; Arndt, Kathleen; Pau, Hans Wilhelm; Podbielski, Andreas

    2012-01-01

    Background High hydrostatic pressure (HHP) treatment can eliminate cholesteatoma cells from explanted human ossicles prior to re-insertion. We analyzed the effects of HHP treatment on the microbial flora on ossicles and on the planktonic and biofilm states of selected isolates. Methodology Twenty-six ossicles were explanted from cholesteatoma patients. Five ossicles were directly analyzed for microbial growth without further treatment. Fifteen ossicles were cut into two pieces. One piece was exposed to HHP of 350 MPa for 10 minutes. Both the treated and untreated (control) pieces were then assessed semi-quantitatively. Three ossicles were cut into two pieces and exposed to identical pressure conditions with or without the addition of one of two different combinations of antibiotics to the medium. Differential effects of 10-minute in vitro exposure of planktonic and biofilm bacteria to pressures of 100 MPa, 250 MPa, 400 MPa and 540 MPa in isotonic and hypotonic media were analyzed using two patient isolates of Staphylococcus epidermidis and Neisseria subflava. Bacterial cell inactivation and biofilm destruction were assessed by colony counting and electron microscopy. Principal Findings A variety of microorganisms were isolated from the ossicles. Irrespective of the medium, HHP treatment at 350 MPa for 10 minutes led to satisfying but incomplete inactivation especially of Gram-negative bacteria. The addition of antibiotics increased the efficacy of elimination. A comparison of HHP treatment of planktonic and biofilm cells showed that the effects of HPP were reduced by about one decadic logarithmic unit when HPP was applied to biofilms. High hydrostatic pressure conditions that are suitable to inactivate cholesteatoma cells fail to completely sterilize ossicles even if antibiotics are added. As a result of the reduced microbial load and the viability loss of surviving bacteria, however, there is a lower risk of re-infection after re-insertion. PMID:22291908

  15. Segmental tuberculosis verrucosa cutis

    Directory of Open Access Journals (Sweden)

    Hanumanthappa H

    1994-01-01

    Full Text Available A case of segmental Tuberculosis Verrucosa Cutis is reported in 10 year old boy. The condition was resembling the ascending lymphangitic type of sporotrichosis. The lesions cleared on treatment with INH 150 mg daily for 6 months.

  16. Chromosome condensation and segmentation

    International Nuclear Information System (INIS)

    Viegas-Pequignot, E.M.

    1981-01-01

    Some aspects of chromosome condensation in mammalians -humans especially- were studied by means of cytogenetic techniques of chromosome banding. Two further approaches were adopted: a study of normal condensation as early as prophase, and an analysis of chromosome segmentation induced by physical (temperature and γ-rays) or chemical agents (base analogues, antibiotics, ...) in order to show out the factors liable to affect condensation. Here 'segmentation' means an abnormal chromosome condensation appearing systematically and being reproducible. The study of normal condensation was made possible by the development of a technique based on cell synchronization by thymidine and giving prophasic and prometaphasic cells. Besides, the possibility of inducing R-banding segmentations on these cells by BrdU (5-bromodeoxyuridine) allowed a much finer analysis of karyotypes. Another technique was developed using 5-ACR (5-azacytidine), it allowed to induce a segmentation similar to the one obtained using BrdU and identify heterochromatic areas rich in G-C bases pairs [fr

  17. International EUREKA: Initialization Segment

    International Nuclear Information System (INIS)

    1982-02-01

    The Initialization Segment creates the starting description of the uranium market. The starting description includes the international boundaries of trade, the geologic provinces, resources, reserves, production, uranium demand forecasts, and existing market transactions. The Initialization Segment is designed to accept information of various degrees of detail, depending on what is known about each region. It must transform this information into a specific data structure required by the Market Segment of the model, filling in gaps in the information through a predetermined sequence of defaults and built in assumptions. A principal function of the Initialization Segment is to create diagnostic messages indicating any inconsistencies in data and explaining which assumptions were used to organize the data base. This permits the user to manipulate the data base until such time the user is satisfied that all the assumptions used are reasonable and that any inconsistencies are resolved in a satisfactory manner

  18. Regulation of EGF and Prostaglandin Expression during Neonatal Gastrointestinal Injury in a Non-Human Primate Explant Model

    Science.gov (United States)

    2017-05-05

    Neonatal Gastrointestinal Injury in a Non-Human Primate Explant Model presented at/published to Pediatric Academic Societies Meeting, San Francisco CA...Medical Center, San Antonio, Texas’ 2Department of Biology, Trinity University, San Antonio, Texas’ JDepartment of Pediatrics /Division of Neonatology

  19. EXPRESIÓN GUS EN EXPLANTES DE Solanum phureja (Juz. et. Buk Var. Criolla Colombia, TRANSFORMADOS CON Agrobacterium tumefaciens

    Directory of Open Access Journals (Sweden)

    IVÁN DARÍO BARRERO-FARFÁN

    2008-01-01

    Full Text Available La expresión transitoria y estable del gen gusA-intron en explantes internodales de papa criolla variedad Criolla Colombia cocultivados con Agrobacterium tumefaciens es reportada. Con el fin de determinar la susceptibilidad de esta variedad a la transformación mediada por A. tumefaciens, explantes internodales de Solanum phureja fueron infectados con la cepa LBA4404 de A. tumefaciens que contiene el plásmido pCAMBIA2301. Este plásmido contiene el gen ntpII que confiere resistencia a kanamicina y el gen reportero gusA-intron. La selección de los explantes potencialmente transgénicos fue realizada en medios con kanamicina. La eficiencia de transformación estable y transitoria fue calculada con base en la actividad GUS (ß-glucuronidasa, detectada por el ensayo histoquímico X-gluc. La expresión transitoria y estable del gen gusA-intron fue observada en células del explante más bien que en tejidos completos. Estos resultados demuestran que la papa criolla (S. phureja Juz. et. Buk variedad Criolla Colombia es susceptible a la infección por A. tumefaciens.

  20. „IN VITRO” EFFECT OF SOME INDUSTRIAL BY-PRODUCTS ON LAVANDULA ANGUSTIFOLIA MILL. EXPLANT GROWTH

    Directory of Open Access Journals (Sweden)

    Corneliu Tanase

    2013-12-01

    Full Text Available After many studies, it was observed that lavender has many therapeutic effects, such as sedation, activities spasmolytic, antiviral, antibacterial. Thus, given the importance of lavender in different areas of human life, in the present study, we studied the influence of natural products bioregulatoars separated from industrial by-products on some lavender stems explants. These explants were inoculated in vitro on MS nutrient media. In these culture media were added polyphenolic extracts obtained from spruce bark and hemp shives, and evaluated their influence on lavender stems explants. The results obtained were compared with those obtained for the control variant, where MS culture medium was used as standard. It was found that the addition of aqueous extract from spruce bark of concentration of 130 mg GAE / L, in the growth of explants of Lavandula angustifolia Mill, an increase in the elongation of the main stem, number of leaves formed, the amount of photoassimilating pigments synthesized and causes the phenomenon of shoots formation. At a higher concentration of the extract (26 mgGAE/100g values are lower.

  1. Interference in adhesion of bacteria and yeasts isolated from explanted voice prostheses to silicone rubber by rhamnolipid biosurfactants

    NARCIS (Netherlands)

    Rodrigues, LR; Banat, IM; van der Mei, HC; Teixeira, JA; Oliveira, R

    Aims: The effects and extent of adhesion of four different bacterial and two yeast strains isolated from explanted voice prostheses to silicone rubber with and without an adsorbed rhamnolipid biosurfactant layer obtained from Pseudomonasaeruginosa DS10-129 was studied. Methods and Results: The

  2. Participation of intracellular cysteine proteinases, in particular cathepsin B, in degradation of collagen in periosteal tissue explants

    NARCIS (Netherlands)

    Creemers, L. B.; Hoeben, K. A.; Jansen, D. C.; Buttle, D. J.; Beertsen, W.; Everts, V.

    1998-01-01

    The involvement of cysteine proteinases in the degradation of soft connective tissue collagen was studied in cultured periosteal explants. Using cysteine proteinase inhibitors that were active intracellularly or extracellularly (Ep453 and Ep475, respectively), it was shown that over-all collagen

  3. METABOLISM AND DNA ADDUCT FORMATION OF 2-ACETYLAMINOFLUORENE BY BLADDER EXPLANTS FROM HUMAN, DOG, MONKEY, HAMSTER AND RAT

    Science.gov (United States)

    It is concluded that bladder explants of the human, dog, monkey, hamster, and rat metabolize AAF mainly to ring-hydroxylated products, but also form small amounts of the proximate carcinogenic metabolite N-hydroxy-AAF. Neither the overall binding of AAF to bladder DNA, nor the fo...

  4. Effect of medium composition and explant size on embryogenic calli formation of cassava (Manihot esculenta Crantz local genotypes

    Directory of Open Access Journals (Sweden)

    ENNY SUDARMONOWATI

    2006-07-01

    Full Text Available Cassava (Manihot esculenta Crantz is an important tropical crop species used for human consumption, feed and raw material for various industries. Genetic transformation through embryogenic tissues is known as an effective method for cassava genetic improvements. Objective of this study was to obtain a suitable medium and length of explants to induce embryogenic callus on friable embryogenic callus (FEC as a target for genetic transformation. Immature leaf lobes (1-3 mm, 3-5 mm and larger than 5 mm in length of local genotypes of cassava (Adira 4. Menti, Iding, Gebang, Rawi and Timtim-29 cultured in vitro were used as explants. The explants were incubated for 2 and 4 weeks on MS (Murashige-Skoog or GD (Greshooff & Doy semi solid medium containing 10 mg/L picloram, 6 mg/L NAA supplemented with 4% sucrose and 4 µM CuSO4. Results showed that the highest percentage (100% of embryogenic calli formation for 4 weeks obtained by culturing Iding of 3-5 mm length on GD semi solid medium, whereas the lowest (33% one obtained by incubation 5 mm leaf lobe of Timtim-29 on the same medium. The most suitable medium for callus induction was GD, whereas the optimum length of explants was 5 mm or larger. Further study needs to be done to obtain friable embryogenic calli (FEC by employing different concentration of picloram and varying other critical factors.

  5. Fluence map segmentation

    International Nuclear Information System (INIS)

    Rosenwald, J.-C.

    2008-01-01

    The lecture addressed the following topics: 'Interpreting' the fluence map; The sequencer; Reasons for difference between desired and actual fluence map; Principle of 'Step and Shoot' segmentation; Large number of solutions for given fluence map; Optimizing 'step and shoot' segmentation; The interdigitation constraint; Main algorithms; Conclusions on segmentation algorithms (static mode); Optimizing intensity levels and monitor units; Sliding window sequencing; Synchronization to avoid the tongue-and-groove effect; Accounting for physical characteristics of MLC; Importance of corrections for leaf transmission and offset; Accounting for MLC mechanical constraints; The 'complexity' factor; Incorporating the sequencing into optimization algorithm; Data transfer to the treatment machine; Interface between R and V and accelerator; and Conclusions on fluence map segmentation (Segmentation is part of the overall inverse planning procedure; 'Step and Shoot' and 'Dynamic' options are available for most TPS (depending on accelerator model; The segmentation phase tends to come into the optimization loop; The physical characteristics of the MLC have a large influence on final dose distribution; The IMRT plans (MU and relative dose distribution) must be carefully validated). (P.A.)

  6. Gamifying Video Object Segmentation.

    Science.gov (United States)

    Spampinato, Concetto; Palazzo, Simone; Giordano, Daniela

    2017-10-01

    Video object segmentation can be considered as one of the most challenging computer vision problems. Indeed, so far, no existing solution is able to effectively deal with the peculiarities of real-world videos, especially in cases of articulated motion and object occlusions; limitations that appear more evident when we compare the performance of automated methods with the human one. However, manually segmenting objects in videos is largely impractical as it requires a lot of time and concentration. To address this problem, in this paper we propose an interactive video object segmentation method, which exploits, on one hand, the capability of humans to identify correctly objects in visual scenes, and on the other hand, the collective human brainpower to solve challenging and large-scale tasks. In particular, our method relies on a game with a purpose to collect human inputs on object locations, followed by an accurate segmentation phase achieved by optimizing an energy function encoding spatial and temporal constraints between object regions as well as human-provided location priors. Performance analysis carried out on complex video benchmarks, and exploiting data provided by over 60 users, demonstrated that our method shows a better trade-off between annotation times and segmentation accuracy than interactive video annotation and automated video object segmentation approaches.

  7. Strategic market segmentation

    Directory of Open Access Journals (Sweden)

    Maričić Branko R.

    2015-01-01

    Full Text Available Strategic planning of marketing activities is the basis of business success in modern business environment. Customers are not homogenous in their preferences and expectations. Formulating an adequate marketing strategy, focused on realization of company's strategic objectives, requires segmented approach to the market that appreciates differences in expectations and preferences of customers. One of significant activities in strategic planning of marketing activities is market segmentation. Strategic planning imposes a need to plan marketing activities according to strategically important segments on the long term basis. At the same time, there is a need to revise and adapt marketing activities on the short term basis. There are number of criteria based on which market segmentation is performed. The paper will consider effectiveness and efficiency of different market segmentation criteria based on empirical research of customer expectations and preferences. The analysis will include traditional criteria and criteria based on behavioral model. The research implications will be analyzed from the perspective of selection of the most adequate market segmentation criteria in strategic planning of marketing activities.

  8. Calogênese e rizogênese em explantes de mogno (Swietenia macrophylla King cultivados in vitro.

    Directory of Open Access Journals (Sweden)

    Silvana Cruz da Rocha

    2010-08-01

    Full Text Available A exploração de árvores tropicais realizada de forma indiscriminada, buscando espécies de alto valor econômico, tem levado várias espécies, como o mogno (Swietenia macrophylla King, ao perigo de extinção. O desenvolvimento de uma metodologia de regeneração de gemas, direta ou indireta, poderia auxiliar na obtenção de um grande número de mudas e constituir uma perspectiva à propagação sexuada. Essa última é limitada pelo fato das sementes perderem rapidamente a capacidade germinativa. No presente trabalho, foram utilizados dois tipos de explantes: fragmentos foliares e de raízes de plantas cultivadas in vitro. Após desinfestação, os explantes foram colocados em meio de cultura de Murashige e Skoog (1962 contendo três quartos da concentração de sais, vitaminas do mesmo meio, 30g.L-1 de sacarose, auxina (ácido naftaleno-acético, ANA, 0,11 µM e 0,54 µM, citocinina (cinetina, CIN, 1,2 µM, 2,3 µM, 4,7 µM e 9,3 µM; 6-benziladenina, BA, 2,2 µM, 4,4 µM e 8,8 µM ou 2-isopenteniladenina, 2-iP, 2,5 µM e 7g.L-1 de ágar. As variáveis testadas foram a concentração e o tipo de regulador de crescimento e a origem dos explantes. A cada 30 dias, os explantes foram avaliados pela contagem do número de explantes formando calos ou raízes e a consistência dos calos. Foram obtidos calos a com base nos dois tipos de explantes. Nos explantes foliares, 90% deles formaram calos em meios de cultura contendo BA 4,4 µM com ANA 0,54 µM e BA 8,9 µM com ANA 0,11 ou 0,54 µM. Nos explantes de raízes, a maior percentagem de explantes com calos foi de 55%, no meio de cultura com BA 2,2 µM e ANA 0,54 µM. Raízes adventícias foram obtidas partindo de calos e do limbo dos explantes foliares, em meios de cultura com CIN e ANA. Não foi observada a formação de gemas adventícias.

  9. Phytochrome A, phytochrome B and HY4 are involved in hypocotyl growth responses to natural radiation in Arabidopsis: weak de-etiolation of the phyA mutant under dense canopies

    International Nuclear Information System (INIS)

    Yanovsky, M.J.; Casal, J.J.; Whitelam, G.C.

    1995-01-01

    The roles of phytochrome A (phyA), phytochrome B (phyB) and a putative blue-light (BL) photoreceptor (HY4) in the control of hypocotyl growth by natural radiation were investigated using phyA, phyB and hy4 mutants of Arabidopsis thaliana. Full sunlight inhibited hypocotyl growth to a larger extent in wild-type (WT) than in phyA, phyB and, particularly, hy4 seedlings. In WT seedlings, hypocotyl growth was promoted by selectively lowering BL irradiance, lowering red-light (R) plus far-red-light (FR) irradiance or lowering the R/FR ratio (which was achieved either by increasing FR or by reducing R). The effects of lowering BL were reduced in hy4 and exaggerated in phyA seedlings. The effects of lowering R+FR were reduced in phyA and exaggerated in hy4 seedlings. Neither phyB nor hy4 mutants responded to low R/FR ratios. Neighbouring plants reflecting FR without shading caused subtle reductions of the R/FR ratio. This signal promoted hypocotyl growth in WT but not in phyA, phyB or hy4 seedlings. Intermediate canopy shade produced similar effects in all genotypes. Under deep shade, de-etiolation was severely impaired in phyA seedlings, which died prematurely. Thus, the FR ‘high-irradiance reaction’ mediated by phyA could be important for seedling survival under dense canopies. (author)

  10. Effects of donor plant age and explants on in vitro culture of Cedrela montana Moritz ex Turcz

    Directory of Open Access Journals (Sweden)

    Sofía Basto

    2012-12-01

    Full Text Available To evaluate the organogenic potential of Cedrela montana Moritz ex Turcz. Explants from mature (10-20 year-old and juvenile (7-18 month-old trees were collected. The first grouping included buds, leaves, and nodes derived from juvenile basal offshoots and rejuvenated shoots from cuttings. The second, included leaves, petioles, nodes, internodes and nodes of in vitro elongated shoots. The highest organogenic potential was observed in nodes from juvenile trees: 45.8% of explants presented axillary bud elongation, while 56.2% presented rooting in a growth regulator free culture medium. Fifty-one percent of elongated shoots produced adventitious shoots with 0.5 μM NAA and 0.5 μM BA; 30% with 0.5 μM NAA and 1 μM BA; and 30% with 1 μM BA. Twenty percent presented roots with 0.5 μM NAA. Root formation was stimulated in a medium supplemented with activated charcoal (5 gL-1. The acclimatization of eighty percent of plantlets regenerated from nodes, and of 72.5% in vitro generated shoots was successful. On the contrary, mature trees material presented low organogenic response. Axillary bud elongation was recorded just in 10.7% of explants from juvenile shoots and in 6.7% of explants from rejuvenated shoots. The age of donor plant and type of explant affect the organogenic potential of C. montana. This study contributes to the understanding of this species’ response under in vitro conditions.

  11. Blockade of Toll-like receptor 2 prevents spontaneous cytokine release from rheumatoid arthritis ex vivo synovial explant cultures

    LENUS (Irish Health Repository)

    Nic An Ultaigh, Sinead

    2011-02-23

    Abstract Introduction The aim of this study was to examine the effect of blocking Toll-like receptor 2 (TLR2) in rheumatoid arthritis (RA) synovial cells. Methods RA synovial tissue biopsies, obtained under direct visualization at arthroscopy, were established as synovial explant cultures ex vivo or snap frozen for immunohistology. Mononuclear cell cultures were isolated from peripheral blood and synovial fluid of RA patients. Cultures were incubated with the TLR1\\/2 ligand, Pam3CSK4 (200 ng, 1 and 10 μg\\/ml), an anti-TLR2 antibody (OPN301, 1 μg\\/ml) or an immunoglobulin G (IgG) (1 μg\\/ml) matched control. The comparative effect of OPN301 and adalimumab (anti-tumour necrosis factor alpha) on spontaneous release of proinflammatory cytokines from RA synovial explants was determined using quantitative cytokine MSD multiplex assays or ELISA. OPN301 penetration into RA synovial tissue explants cultures was assessed by immunohistology. Results Pam3CSK4 significantly upregulated interleukin (IL)-6 and IL-8 in RA peripheral blood mononuclear cells (PBMCs), RA synovial fluid mononuclear cells (SFMCs) and RA synovial explant cultures (P < 0.05). OPN301 significantly decreased Pam3CSK4-induced cytokine production of tumour necrosis factor alpha (TNF-α), IL-1β, IL-6, interferon (IFN)-γ and IL-8 compared to IgG control in RA PBMCs and SFMCs cultures (all P < 0.05). OPN301 penetration of RA synovial tissue cultures was detected in the lining layer and perivascular regions. OPN301 significantly decreased spontaneous cytokine production of TNF-α, IL-1β, IFN-γ and IL-8 from RA synovial tissue explant cultures (all P < 0.05). Importantly, the inhibitory effect of OPN on spontaneous cytokine secretion was comparable to inhibition by anti-TNFα monoclonal antibody adalimumab. Conclusions These findings further support targeting TLR2 as a potential therapeutic agent for the treatment of RA.

  12. Segmented block copolymers with monodisperse aramide end-segments

    NARCIS (Netherlands)

    Araichimani, A.; Gaymans, R.J.

    2008-01-01

    Segmented block copolymers were synthesized using monodisperse diaramide (TT) as hard segments and PTMO with a molecular weight of 2 900 g · mol-1 as soft segments. The aramide: PTMO segment ratio was increased from 1:1 to 2:1 thereby changing the structure from a high molecular weight multi-block

  13. Rediscovering market segmentation.

    Science.gov (United States)

    Yankelovich, Daniel; Meer, David

    2006-02-01

    In 1964, Daniel Yankelovich introduced in the pages of HBR the concept of nondemographic segmentation, by which he meant the classification of consumers according to criteria other than age, residence, income, and such. The predictive power of marketing studies based on demographics was no longer strong enough to serve as a basis for marketing strategy, he argued. Buying patterns had become far better guides to consumers' future purchases. In addition, properly constructed nondemographic segmentations could help companies determine which products to develop, which distribution channels to sell them in, how much to charge for them, and how to advertise them. But more than 40 years later, nondemographic segmentation has become just as unenlightening as demographic segmentation had been. Today, the technique is used almost exclusively to fulfill the needs of advertising, which it serves mainly by populating commercials with characters that viewers can identify with. It is true that psychographic types like "High-Tech Harry" and "Joe Six-Pack" may capture some truth about real people's lifestyles, attitudes, self-image, and aspirations. But they are no better than demographics at predicting purchase behavior. Thus they give corporate decision makers very little idea of how to keep customers or capture new ones. Now, Daniel Yankelovich returns to these pages, with consultant David Meer, to argue the case for a broad view of nondemographic segmentation. They describe the elements of a smart segmentation strategy, explaining how segmentations meant to strengthen brand identity differ from those capable of telling a company which markets it should enter and what goods to make. And they introduce their "gravity of decision spectrum", a tool that focuses on the form of consumer behavior that should be of the greatest interest to marketers--the importance that consumers place on a product or product category.

  14. Scorpion image segmentation system

    Science.gov (United States)

    Joseph, E.; Aibinu, A. M.; Sadiq, B. A.; Bello Salau, H.; Salami, M. J. E.

    2013-12-01

    Death as a result of scorpion sting has been a major public health problem in developing countries. Despite the high rate of death as a result of scorpion sting, little report exists in literature of intelligent device and system for automatic detection of scorpion. This paper proposed a digital image processing approach based on the floresencing characteristics of Scorpion under Ultra-violet (UV) light for automatic detection and identification of scorpion. The acquired UV-based images undergo pre-processing to equalize uneven illumination and colour space channel separation. The extracted channels are then segmented into two non-overlapping classes. It has been observed that simple thresholding of the green channel of the acquired RGB UV-based image is sufficient for segmenting Scorpion from other background components in the acquired image. Two approaches to image segmentation have also been proposed in this work, namely, the simple average segmentation technique and K-means image segmentation. The proposed algorithm has been tested on over 40 UV scorpion images obtained from different part of the world and results obtained show an average accuracy of 97.7% in correctly classifying the pixel into two non-overlapping clusters. The proposed 1system will eliminate the problem associated with some of the existing manual approaches presently in use for scorpion detection.

  15. Nicotiana plumbaginifolia hlg mutants have a mutation in a PHYB-type phytochrome gene: they have elongated hypocotyls in red light, but are not elongated as adult plants.

    Science.gov (United States)

    Hudson, M; Robson, P R; Kraepiel, Y; Caboche, M; Smith, H

    1997-11-01

    Two new allelic mutants of Nicotiana plumbaginifolia have been isolated which display a hypocotyl which is long (hlg) when seedlings are grown in continuous white light (W). This can be accounted for by the decreased response to red light (R) of the hypocotyl elongation rate in these mutants. Responses to other wavelengths are unaffected in the mutants. When grown in white light, mature hlg mutants are not elongated with respect to the wild-type; they also bolt and flower later. The shade-avoidance responses to red/far red ratio (R:FR) are intact in these mutants. Both mutants are deficient in phyB-like polypeptide that is immunodetectable in the wild-type; both have wild-type levels of a phyA-like polypeptide. These alleles are inherited in a partially dominant manner, and correspond to single-base missense mutations in a gene highly homologous to N. tabacum PHYB, which codes for a phytochrome B-type photoreceptor. One allele, hlg-1, has an introduced amino acid substitution; this may define a residue essential for phytochrome protein stability. The other allele, hlg-2, has a stop codon introduced C-terminal to the chromophore binding domain. As these phyB mutants are unaffected in shade-avoidance responses, but deficient in perception of R, it is concluded that the phyB absent in these mutants is responsible for R perception in the N. plumbaginifolia seedling, but is not a R:FR sensor in light-grown plants.

  16. Genome-wide identification and characterization of miRNAs in the hypocotyl and cotyledon of cauliflower (Brassica oleracea L. var. botrytis) seedlings.

    Science.gov (United States)

    Geng, Meijuan; Li, Hui; Jin, Chuan; Liu, Qian; Chen, Chengbin; Song, Wenqin; Wang, Chunguo

    2014-02-01

    MicroRNAs (miRNAs) are a class of small endogenous, non-coding RNAs that have key regulatory functions in plant growth, development, and other biological processes. Hypocotyl and cotyledon are the two major tissues of cauliflower (Brassica oleracea L. var. botrytis) seedlings. Tissue culture experiments have indicated that the regenerative abilities of these two tissues are significantly different. However, the characterization of miRNAs and their roles in regulating organ development in cauliflower remain unexplored. In the present study, two small RNA libraries were sequenced by Solexa sequencing technology. 99 known miRNAs belonging to 28 miRNA families were identified, in which 6 miRNA families were detected only in Brassicaceae. A total of 162 new miRNA sequences with single nucleotide substitutions corresponding to the known miRNAs, and 32 potentially novel miRNAs were also first discovered. Comparative analysis indicated that 42 of 99 known miRNAs and 17 of 32 novel miRNAs exhibited significantly differential expression between hypocotyl and cotyledon, and the differential expression of several miRNAs was further validated by stem-loop RT-PCR. In addition, 235 targets for 89 known miRNAs and 198 targets for 24 novel miRNAs were predicted, and their functions were further discussed. The expression patterns of several representative targets were also confirmed by qRT-PCR analysis. The results identified that the transcriptional expression patterns of miRNAs were negatively correlated with their targets. These findings gave new insights into the characteristics of miRNAs in cauliflower, and provided important clues to elucidate the roles of miRNAs in the tissue differentiation and development of cauliflower.

  17. Segmentation of complex document

    Directory of Open Access Journals (Sweden)

    Souad Oudjemia

    2014-06-01

    Full Text Available In this paper we present a method for segmentation of documents image with complex structure. This technique based on GLCM (Grey Level Co-occurrence Matrix used to segment this type of document in three regions namely, 'graphics', 'background' and 'text'. Very briefly, this method is to divide the document image, in block size chosen after a series of tests and then applying the co-occurrence matrix to each block in order to extract five textural parameters which are energy, entropy, the sum entropy, difference entropy and standard deviation. These parameters are then used to classify the image into three regions using the k-means algorithm; the last step of segmentation is obtained by grouping connected pixels. Two performance measurements are performed for both graphics and text zones; we have obtained a classification rate of 98.3% and a Misclassification rate of 1.79%.

  18. Connecting textual segments

    DEFF Research Database (Denmark)

    Brügger, Niels

    2017-01-01

    history than just the years of the emergence of the web, the chapter traces the history of how segments of text have deliberately been connected to each other by the use of specific textual and media features, from clay tablets, manuscripts on parchment, and print, among others, to hyperlinks on stand......In “Connecting textual segments: A brief history of the web hyperlink” Niels Brügger investigates the history of one of the most fundamental features of the web: the hyperlink. Based on the argument that the web hyperlink is best understood if it is seen as another step in a much longer and broader...

  19. Effect of environmental and cultural conditions on medium pH and explant growth performance of Douglas-fir (Pseudotsuga menziesii shoot cultures [version 2; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Chien-Chih Chen

    2015-05-01

    Full Text Available The medium pH level of plant tissue cultures has been shown to be essential to many aspects of explant development and growth. Sensitivity or tolerance of medium pH change in vitro varies according to specific requirements of individual species. The objectives of this study are to 1 determine medium pH change over time in storage conditions and with presence of explants, 2 evaluate the effects of medium pH change on explant growth performance and 3 assess the effects of adding a pH stabilizer, 2-(N-morpholinoethanesulfonic acid (MES that is commonly used in Douglas-fir micropropagation medium. Vegetative buds were collected in the spring before breaking dormancy from juvenile and mature donor trees for conducting these evaluations. Medium, with or without MES, was pre-adjusted to five pH levels before adding MES, agar and autoclaving. Medium pH changes and explant growth parameters were measured at eight different incubation times. Overall, MES provided a more stable medium pH, relative to starting pH values, under both light and dark storage conditions as well as with presence of explants. A general trend of decreasing medium pH over time was found comparing explants from juvenile and mature donor genotypes. Explant height and weight growth increased over time, but differ among explants from juvenile and mature donor genotypes. Our findings suggest that a 21-day subculture practice may best sustain medium freshness, medium pH level and desirable explant growth.

  20. Fibroid explants reveal a higher sensitivity against MDM2-inhibitor nutlin-3 than matching myometrium

    Directory of Open Access Journals (Sweden)

    Markowski Dominique N

    2012-01-01

    Full Text Available Abstract Background Spontaneous cessation of growth is a frequent finding in uterine fibroids. Increasing evidence suggests an important role of cellular senescence in this growth control. Deciphering the underlying mechanisms of growth control that can be expected not only to shed light on the biology of the tumors but also to identify novel therapeutic targets. Methods We have analyzed uterine leiomyomas and matching normal tissue for the expression of p14Arf and used explants to see if reducing the MDM2 activity using the small-molecule inhibitor nutlin-3 can induce p53 and activate genes involved in senescence and/or apoptosis. For these studies quantitative real-time RT-PCR, Western blots, and immunohistochemistry were used. Statistical analyses were performed using the student's t test. Results An in depth analysis of 52 fibroids along with matching myometrium from 31 patients revealed in almost all cases a higher expression of p14Arf in the tumors than in the matching normal tissue. In tissue explants, treatment with the MDM2 inhibitor nutlin-3 induced apoptosis as well as senescence as revealed by a dose-dependent increase of the expression of BAX as well as of p21, respectively. Simultaneously, the expression of the proliferation marker Ki-67 drastically decreased. Western-blot analysis identified an increase of the p53 level as the most likely reason for the increased activity of its downstream markers BAX and p21. Because as a rule fibroids express much higher levels of p14Arf, a major negative regulator of MDM2, than matching myometrium it was then analyzed if fibroids are more sensitive against nutlin-3 treatment than matching myometrium. We were able to show that in most fibroids analyzed a higher sensibility than that of matching myometrium was noted with a corresponding increase of the p53 immunopositivity of the fibroid samples compared to those from myometrium. Conclusions The results show that uterine fibroids represent a cell

  1. Effect of environmental and cultural conditions on medium pH and explant growth performance of Douglas-fir ( Pseudotsuga menziesii) shoot cultures

    OpenAIRE

    Chen, Chien-Chih; Bates, Rick; Carlson, John

    2015-01-01

    The medium pH level of plant tissue cultures has been shown to be essential to many aspects of explant development and growth. Sensitivity or tolerance of medium pH change in vitro varies according to specific requirements of individual species. The objectives of this study are to 1) determine medium pH change over time in storage conditions and with presence of explants, 2) evaluate the effects of medium pH change on explant growth performance and 3) assess the effects of adding a pH stabili...

  2. Segmentation in cinema perception.

    Science.gov (United States)

    Carroll, J M; Bever, T G

    1976-03-12

    Viewers perceptually segment moving picture sequences into their cinematically defined units: excerpts that follow short film sequences are recognized faster when the excerpt originally came after a structural cinematic break (a cut or change in the action) than when it originally came before the break.

  3. Dictionary Based Image Segmentation

    DEFF Research Database (Denmark)

    Dahl, Anders Bjorholm; Dahl, Vedrana Andersen

    2015-01-01

    We propose a method for weakly supervised segmentation of natural images, which may contain both textured or non-textured regions. Our texture representation is based on a dictionary of image patches. To divide an image into separated regions with similar texture we use an implicit level sets...

  4. Unsupervised Image Segmentation

    Czech Academy of Sciences Publication Activity Database

    Haindl, Michal; Mikeš, Stanislav

    2014-01-01

    Roč. 36, č. 4 (2014), s. 23-23 R&D Projects: GA ČR(CZ) GA14-10911S Institutional support: RVO:67985556 Keywords : unsupervised image segmentation Subject RIV: BD - Theory of Information http://library.utia.cas.cz/separaty/2014/RO/haindl-0434412.pdf

  5. Metrics for image segmentation

    Science.gov (United States)

    Rees, Gareth; Greenway, Phil; Morray, Denise

    1998-07-01

    An important challenge in mapping image-processing techniques onto applications is the lack of quantitative performance measures. From a systems engineering perspective these are essential if system level requirements are to be decomposed into sub-system requirements which can be understood in terms of algorithm selection and performance optimization. Nowhere in computer vision is this more evident than in the area of image segmentation. This is a vigorous and innovative research activity, but even after nearly two decades of progress, it remains almost impossible to answer the question 'what would the performance of this segmentation algorithm be under these new conditions?' To begin to address this shortcoming, we have devised a well-principled metric for assessing the relative performance of two segmentation algorithms. This allows meaningful objective comparisons to be made between their outputs. It also estimates the absolute performance of an algorithm given ground truth. Our approach is an information theoretic one. In this paper, we describe the theory and motivation of our method, and present practical results obtained from a range of state of the art segmentation methods. We demonstrate that it is possible to measure the objective performance of these algorithms, and to use the information so gained to provide clues about how their performance might be improved.

  6. ORGANOGÊNESE IN VITRO DE Citrus EM FUNÇÃO DE CONCENTRAÇÕES DE BAP E SECCIONAMENTO DO EXPLANTE CITRUS IN VITRO ORGANOGENESIS RELATED TO BAP CONCENTRATIONS AND EXPLANT SECTION

    Directory of Open Access Journals (Sweden)

    THAÍS LACAVA DE MOURA

    2001-08-01

    Full Text Available O sucesso de técnicas biotecnológicas no melhoramento in vitro de Citrus depende diretamente do desenvolvimento de protocolos eficientes para regeneração de plantas. Objetivou-se avaliar o efeito de concentrações de 6-benzilaminopuria (BAP na organogênese in vitro de limão-'Cravo' e laranja-'Pêra', bem como o efeito do seccionamento do explante em laranja-'Valência'. Para o limão-'Cravo', foram utilizados como explante, segmentos internodais de plântulas germinadas in vitro, cultivados em meio MT e variando-se as concentrações de BAP em 0; 2,5; 5; 7,5 e 10 mg.L-1. Nas laranjas-'Pêra' e 'Valência' os explantes foram segmentos do epicótilo de plântulas germinadas in vitro. Os explantes de laranja-'Pêra' foram cultivados em meio MT variando-se as concentrações de BAP em 0; 1; 2; 3 e 4 mg.L-1. Para a laranja-'Valência', metade dos explantes foram seccionados e cultivados em meio MT acrescido de 1,0 mg.L-1 de BAP. Todas as brotações obtidas foram alongadas no meio de cultura MT + 25 g.L-1 de sacarose + 1 mg.L-1 de ácido giberélico (GA3 e enraizadas no meio MT + 25 g.L-1 de sacarose + 0,5 g.L-1 de carvão ativado + 1 mg.L-1 de ácido naftaleno acético (ANA. O melhor resultado para o número de brotações adventícias foi obtido na concentração 2,5 mg.L-1 de BAP para limão-'Cravo', e nas concentrações 1,0 e 2,0 mg.L-1 de BAP para laranja-'Pêra'. O seccionamento dos explantes favoreceu a organogênese in vitro da laranja-'Valência', porém as brotações apresentaram menor índice de enraizamento.The establishment of efficient plant regeneration protocols is essential for the success and application of in vitro breeding biotechnologies in Citrus. The objective of this work was to verify the effect of 6-benzilaminopurine (BAP on the in vitro organogenesis of Rangpur lime (Citrus limonia (L. Osbeck and 'Pera' sweet orange (Citrus sinensis (L. Osbeck, and the effect of cutting the explant on the in vitro organogenesis of

  7. Single leg separation prevalence among explanted Björk-Shiley prosthetic heart valves.

    Science.gov (United States)

    Blot, William J; Signorello, Lisa B; Cohen, Sarah S; Ibrahim, Michel A

    2007-11-01

    Björk-Shiley convexo-concave (BSCC) prosthetic heart valves are believed to have been implanted in over 86,000 patients worldwide. Limited data are available on the prevalence of single leg separations (SLS) of the valves' outlet struts, a potential precursor to complete valve fracture. Data maintained by the manufacturer, including results of examinations for SLS in explanted valves, were merged with available information on the characteristics of the valve. The prevalence of SLS in the examined valves was calculated according to valve angle, size, position, and study. Among 343 examined valves, the overall prevalence of SLS was 8.2%, but this varied significantly by valve size, being three-fold higher among 29+ mm valves than among smaller valves, with statistically non-significantly higher prevalences among mitral than aortic, and among 70 degrees than 60 degrees valves. By applying the size, position and angle-specific SLS prevalences to the worldwide valve distribution, it is estimated that SLS may be present in 6.8% (95% confidence limits 4.1-9.4%) of all BSCC valves. These findings suggest that SLS may affect between 820 and 1,880 of the almost 20,000 BSCC valves among surviving patients worldwide. Such estimates help frame the context for potential patient screenings, should imaging and acoustic techniques to detect SLS become available.

  8. High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation

    Science.gov (United States)

    Clutterbuck, Abigail L.; Smith, Julia R.; Allaway, David; Harris, Pat; Liddell, Susan; Mobasheri, Ali

    2011-01-01

    This study employed a targeted high-throughput proteomic approach to identify the major proteins present in the secretome of articular cartilage. Explants from equine metacarpophalangeal joints were incubated alone or with interleukin-1beta (IL-1β, 10 ng/ml), with or without carprofen, a non-steroidal anti-inflammatory drug, for six days. After tryptic digestion of culture medium supernatants, resulting peptides were separated by HPLC and detected in a Bruker amaZon ion trap instrument. The five most abundant peptides in each MS scan were fragmented and the fragmentation patterns compared to mammalian entries in the Swiss-Prot database, using the Mascot search engine. Tryptic peptides originating from aggrecan core protein, cartilage oligomeric matrix protein (COMP), fibronectin, fibromodulin, thrombospondin-1 (TSP-1), clusterin (CLU), cartilage intermediate layer protein-1 (CILP-1), chondroadherin (CHAD) and matrix metalloproteinases MMP-1 and MMP-3 were detected. Quantitative western blotting confirmed the presence of CILP-1, CLU, MMP-1, MMP-3 and TSP-1. Treatment with IL-1β increased MMP-1, MMP-3 and TSP-1 and decreased the CLU precursor but did not affect CILP-1 and CLU levels. Many of the proteins identified have well-established extracellular matrix functions and are involved in early repair/stress responses in cartilage. This high throughput approach may be used to study the changes that occur in the early stages of osteoarthritis. PMID:21354348

  9. Shoot Organogenesis and Plant Regeneration from Leaf Explants of Lysionotus serratus D. Don

    Directory of Open Access Journals (Sweden)

    Qiansheng Li

    2013-01-01

    Full Text Available The gesneriaceous perennial plant, Lysionotus serratus, has been used in traditional Chinese medicine. It also has a great development potential as an ornamental plant with its attractive foliage and beautiful flowers. An efficient propagation and regeneration system via direct shoot organogenesis from leaf explant was established in this study. High active cytokinin (6-benzyladenine (BA or thidiazuron (TDZ was effective for direct organogenesis of initial induction. Murashige and Skoog (MS growth media containing 0.5 mg L−1 BA alone or with combination of 0.1 mg L−1  α-Naphthaleneacetic acid (NAA were the most effective for shoot proliferation. High BA concentration (1.0 mg L−1 in the media caused high percentage of vitrified shoots though they introduced high shoot proliferation rate. Histological observation indicated that adventitious shoot regeneration on the medium containing 0.5 mg L−1 BA alone occurred directly from leaf epidermal cells without callus formation. Regenerated shoots rooted well on medium containing half-strength MS medium with 0.5 mg L−1 indole-3-butyric acid (IBA and indole-3-acetic acid (IAA, and the plantlets successfully acclimatized and grew vigorously in the greenhouse with a 94.2% and 92.1% survival rate.

  10. Morphogenic responses of three explants of Lupinus montanus (H.B.K. cultured in vitro

    Directory of Open Access Journals (Sweden)

    Gabriel Ramírez-González

    2015-01-01

    Full Text Available Resumen El necrosamiento y la respuesta morfogénica de explantes de cotiledón, epicótilo e hipocótilo de plántulas de Lupinus montanus (H.B.K. germinadas in vitro se evaluaron para establecer las condiciones básicas de micropropagación. El necrosamiento se evaluó en medio MS con 0.40 mg·litro-1 de tiamina, 100 mg·litro-1 de mioinositol, 3 % de sacarosa y 7 g·litro-1 de agar-agar y distintas dosis de carbón activado (CA y ácido cítrico. Los tratamientos con CA (50 mg·litro-1 y 100 mg·litro-1 fueron los mejores en el control del necrosamiento (P = 0.001; el cotiledón tuvo mayor nivel de necrosamiento ( 70 % a diferencia del cotiledón, cuya respuesta fue primordialmente callogénica (50 %. El epicótilo cultivado en medio MS con 3.0 μM de AIA y 1.0 μM de BA tuvo el mayor número de brotes (10 y altura (11.4 ± 2.6 cm.

  11. In vitro neurotoxic effects of 1 GeV/n iron particles assessed in retinal explants.

    Science.gov (United States)

    Vazquez, M E; Kirk, E

    2000-01-01

    The heavy ion component of the cosmic radiation remains problematic to the assessment of risk in manned space flight. The biological effectiveness of HZE particles has yet to be established, particularly with regard to nervous tissue. Using heavy ions accelerated at the AGS of Brookhaven National Laboratory, we study the neurotoxic effects of iron particles. We exposed retinal explants, taken from chick embryos, to determine the dose response relationships for neurite outgrowth. Morphometric techniques were used to evaluate the in vitro effects of 1 GeV/a iron particles (LET 148 keV/micrometer). Iron particles produced a dose-dependent reduction of neurite outgrowth with a maximal effect achieved with a dose of 100 cGy. Doses as low as 10-50 cGy were able to induce reductions of the neurite outgrowth as compared to the control group. Neurite generation is a more sensitive parameter than neurite elongation, suggesting different mechanism of radiation damage in our model. These results showed that low doses/fluences of iron particles could impair the retinal ganglion cells' capacity to generate neurites indicating the highly neurotoxic capability of this heavy charged particle.

  12. Assessment of the photoprotection properties of sunscreens by chromatographic measurement of DNA damage in skin explants.

    Science.gov (United States)

    Mouret, Stéphane; Bogdanowicz, Patrick; Haure, Marie-José; Castex-Rizzi, Nathalie; Cadet, Jean; Favier, Alain; Douki, Thierry

    2011-01-01

    Evaluation of the photoprotection provided by sunscreens is performed either through the induction of erythema and expressed as the sun protection factor (SPF), or by the UVA-mediated persistent pigment darkening (PPD). None of these two endpoints has a link with skin cancer, the most deleterious consequence of excess exposure to solar UV radiation. We thus set up a complementary approach to evaluate the protection provided by sunscreens to the genome of human skin. This is based on the quantification of the thymine cyclobutane dimer (TT-CPD), the main DNA lesion induced by both UVB and UVA radiations. Irradiations were performed ex vivo on human skin explants and the level of TT-CPD in DNA was determined by HPLC associated with tandem mass spectrometry. The technique was first optimized and validated with three standard sunscreens. The study was then extended to the evaluation of a commercial high SPF sunscreen exhibiting efficient UVA photoprotection. The DNA protecting factor was found to reflect the ratio between UVB and UVA photoprotection, although the absolute values of the genomic protection were, as a general trend, lower than either SPF or PPD. These data show the usefulness of the proposed approach for the evaluation of the genoprotection afforded by sunscreens. © 2010 The Authors. Photochemistry and Photobiology © 2010 The American Society of Photobiology.

  13. Induction of shoot regeneration in cotyledon explants of the oilseed crop Sesamum indicum L.

    Directory of Open Access Journals (Sweden)

    Masochon Zimik

    2017-12-01

    Full Text Available Sesamum indicum is an ancient oilseed crop known for its high quality edible oil and its medicinally important lignans. The crop is said to be recalcitrant to plant tissue culture thus limiting the use of modern biotechnology for its genetic improvement. We present here a protocol describing plant regeneration through adventitious shoot formation from cotyledons dissected from sesame seeds soaked for four hours in water. Subculturing of the cotyledons after two weeks of culture on to a fresh Murashige and Skoog medium leads to differentiation of adventitious shoots from the proximal cut end of the explant. Culture of cotyledons on a medium containing 9% sucrose for a couple of weeks prior to transfer to MS medium supplemented with 3% sucrose induced a higher frequency of shoot regeneration. The highest frequency of 25% adventitious shoot regeneration was observed for S. indicum variety UMA. This variety also turned out to be the best among the ten genotypes tested for shoot regeneration through tissue culture. While addition of IAA marginally improved regeneration, silver nitrate was found essential for enhancing the frequency of shoot regeneration. The regenerated shoots formed roots on full strength MS medium supplemented with 1 mg/l IBA and the rooted plants were established in soil.

  14. Analysis of explanted ePTFE cardiovascular grafts (modified BT shunt)

    Energy Technology Data Exchange (ETDEWEB)

    Doble, Mukesh [Department of Biotechnology, Indian Institute of Technology, Chennai 600 036 (India); Makadia, Nilesh; Pavithran, Sreeja; Kumar, R Suresh [Department of Pediatric Cardiology, Institute of Cardio-Vascular Diseases, Madras Medical Mission, Chennai, 600 037 (India)], E-mail: mukeshd@iitm.ac.in

    2008-09-01

    Structural, chemical, mechanical and surface changes were studied in expanded polytetrafluroethylene vascular grafts explanted from children undergoing planned surgical management of congenital heart disease. These grafts were implanted when recipients were aged 7 days to 8 years (median-48 weeks) and they had been in circulation for a period of 10-52 months (median-74 weeks). While no chemical changes were observed in the shunt, on average the tensile strength had decreased by 50%, total elongation by 61% and crystallinity by 3%. No salt deposits were observed on the surface of the graft. Soluble and insoluble proteins were bound to the polymer surface, which had made the surface hydrophilic. The external surface roughness had increased by 254.5 and the internal surface roughness by 2.6 times the initial value. The fine polymer structure had become fused and clumped. The fusing of strands on the polymer surface became more pronounced with longer duration of implantation. In one instance of previously documented graft stenosis, the heat capacity was found to be more than that of the unimplanted sample, indicating an increase in crystallinity. A longer period of study with a larger sample size would likely shed more light on the relation between physico-chemical changes and graft stenosis.

  15. Analysis of explanted ePTFE cardiovascular grafts (modified BT shunt)

    International Nuclear Information System (INIS)

    Doble, Mukesh; Makadia, Nilesh; Pavithran, Sreeja; Kumar, R Suresh

    2008-01-01

    Structural, chemical, mechanical and surface changes were studied in expanded polytetrafluroethylene vascular grafts explanted from children undergoing planned surgical management of congenital heart disease. These grafts were implanted when recipients were aged 7 days to 8 years (median-48 weeks) and they had been in circulation for a period of 10-52 months (median-74 weeks). While no chemical changes were observed in the shunt, on average the tensile strength had decreased by 50%, total elongation by 61% and crystallinity by 3%. No salt deposits were observed on the surface of the graft. Soluble and insoluble proteins were bound to the polymer surface, which had made the surface hydrophilic. The external surface roughness had increased by 254.5 and the internal surface roughness by 2.6 times the initial value. The fine polymer structure had become fused and clumped. The fusing of strands on the polymer surface became more pronounced with longer duration of implantation. In one instance of previously documented graft stenosis, the heat capacity was found to be more than that of the unimplanted sample, indicating an increase in crystallinity. A longer period of study with a larger sample size would likely shed more light on the relation between physico-chemical changes and graft stenosis

  16. Culturated rat cerebral cortex explants and their application in the study of SPECT scan radiopharaceuticals

    International Nuclear Information System (INIS)

    Jong, B.M. de.

    1989-01-01

    In this thesis mechanics that result in the distinct localization of radiopharmaceuticals within the brain have been investigated. In order to 'get more insight' in uptake and binding of radiopharmaceuticals bu brain tissue, use has been made of the tissue culture technique. Tissue culture privides the opportunity of doing experiments with brain tissue under stable conditions, in the absence of a blood-brain barrier, and without interference by cerebral blood flow. The present thesis is presented in two sections. The first part focusses on longterm culture of 'organotypic' cerebral neocortex tissue, obtained from neonatal rat brain and explanted into a chemically defined medium. Procedures were developed which enabled culturing of this tissue without the occurence of central necrosis and with the preservation of a characteristic histiotypic organization. Morphological characteristics of the cultures were described and measured at various ages in vitro. In the second part, the cultures were used to study mechanisms that might contribute to the tissue uptake of radiopharmaceuticals which are in clinical use for SPECT brain imaging. (author). 369 refs.; 50 figs.; 13 tabs

  17. Changes of free, soluble conjugated and bound polyamine titers of jojoba explants under sodium chloride salinity in vitro.

    Science.gov (United States)

    Roussos, Peter A; Pontikis, Constantine A

    2007-07-01

    Jojoba (Simmondsia chinensis L.) single node explants were cultured in a basal medium supplemented with 17.8 microM 6-benzyladenine and four levels of sodium chloride concentration (0, 56.41, 112.82 and 169.23 mM). The free, the soluble conjugated and the insoluble bound forms of polyamines (PAs) (putrescine (Put), spermidine (Spd) and spermine (Spm)) were determined monthly during a 3-month proliferation stage. Free Put and Spd were found in higher levels in the control treatment, while Spm content was higher in the salt treatments. All soluble conjugated PAs were found to be in lower concentrations in explants growing on medium supplemented with salt, while the opposite was true for the insoluble bound PAs. It appeared that certain PAs and PAs forms could play a significant role in the adaptation mechanism of jojoba under saline conditions.

  18. The effect of protease inhibitors on the induction of osteoarthritis-related biomarkers in bovine full-depth cartilage explants

    DEFF Research Database (Denmark)

    He, Yi; Zheng, Qinlong; Jiang, Mengmeng

    2015-01-01

    contribution of ADAMTS-4 and ADAMTS-5 to cartilage degradation upon catabolic stimulation; ii) To investigate the effect of regulating the activities of key enzymes by mean of broad-spectrum inhibitors. Methods Bovine full-depth cartilage explants stimulated with tumor necrosis factor alpha (TNF...... protease for the generation of 374ARGS aggrecan fragment in the TNF-α/OSM stimulated bovine cartilage explants. This study addresses the need to determine the roles of ADAMTS-4 and ADAMTS-5 in human articular degradation in OA and hence identify the attractive target for slowing down human cartilage......Objective The specific degradation of type II collagen and aggrecan by matrix metalloproteinase (MMP)-9, -13 and ADAMTS-4 and -5 (aggrecanase-1 and -2) in the cartilage matrix is a critical step in pathology of osteoarthritis (OA). The aims of this study were: i) To investigate the relative...

  19. Agrobacterium-mediated transformation of Citrus sinensis and Citrus limonia epicotyl segments Transformação genética em Citrus sinensis e Citrus limonia mediada por Agrobacterium tumefaciens a partir de segmentos de epicótilo

    Directory of Open Access Journals (Sweden)

    Weliton Antonio Bastos de Almeida

    2003-02-01

    Full Text Available Genetic transformation allows the release of improved cultivars with desirable characteristics in a shorter period of time and therefore may be useful in citrus breeding programs. The objective of this research was to establish a protocol for genetic transformation of Valencia and Natal sweet oranges (Citrus sinensis L. Osbeck and Rangpur lime (Citrus limonia L. Osbeck. Epicotyl segments of germinated in vitro plantlets (three weeks in darkness and two weeks in a 16-h photoperiod were used as explants. These were co-cultivated with Agrobacterium tumefaciens strain EHA-105 and different experiments were done to evaluate the transformation efficiency: explants were co-cultivated with Agrobacterium for one, three or five days; explants were incubated with Agrobacterium suspension for 5, 10, 20 or 40 minutes; co-cultivation medium was supplemented with acetosyringone at 0, 100 or 200 µmol L-1; Explants ends had a longitudinal terminal incision (2-3 mm; co-cultivation temperatures of 19, 23 or 27°C were imposed. The experimental design was completely randomized in all experiments with five replications, each consisted of a Petri dish (100 x 15 mm with 30 explants and resulted in a total of 150 explants per treatment. Longitudinal terminal incision in the explant ends did not improve shoot regeneration. However, transgenic plants of all three cultivars were confirmed from explants that had been subjected to inoculation time of 20 minutes, co-culture of three days at 23-27°C, in the absence of acetosyringone.A transformação genética permite produzir cultivares com características específicas e pode, dessa forma ser associada a programas de melhoramento de citros. O objetivo deste trabalho foi estabelecer protocolos de transformação genética para as laranjas doce 'Valência' e 'Natal' (Citrus sinensis L. Osbeck, bem como para o limão 'Cravo'(Citrus limonia L. Osbeck. Segmentos de epicótilo de plântulas germinadas in vitro (três semanas no

  20. Effect of gamma radiation on different explants of sweet potato (Ipomoea batatas L. (Lam)) to induce NaCl tolerance

    International Nuclear Information System (INIS)

    Gutierrez-Rosati, A.

    1997-01-01

    Clonal lines tolerant to NaCl were obtained by combining in vitro culture and gamma radiation in two Peruvian varieties 'Amarillo de Quillabamba' and 'Nemanete'. The most suitable explants were pedicel sections and leaf blades. Embryogenic callus was induced on basal MS basal medium containing 0.5 ppm 2,4-D. The embryogenic calli were irradiated with 5 Gy from a 137 Cs source. Several putative mutants appeared to be stable. (author). 9 refs, 3 tabs

  1. Plant regeneration of non-toxic Jatropha curcas—impacts of plant growth regulators, source and type of explants

    KAUST Repository

    Kumar, Nitish

    2011-01-28

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel plant, however, oil and deoiled cake are toxic. A non-toxic variety of J. curcas is reported from Mexico. The present investigation explores the effects of different plant growth regulators (PGRs) viz. 6-benzyl aminopurine (BAP) or thidiazuron (TDZ) individually and in combination with indole-3-butyric acid (IBA), on regeneration from in vitro and field-grown mature leaf explants, in vitro and glasshouse-grown seedlings cotyledonary leaf explants of non-toxic J. curcas. In all the tested parameters maximum regeneration efficiency (81.07%) and the number of shoot buds per explants (20.17) was observed on 9.08 μM TDZ containing Murashige and Skoog’s (MS) medium from in vitro cotyledonary leaf explants. The regenerated shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with 2.25 μM BAP and 8.5 μM IAA. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing different concentrations and combinations of IBA, IAA and NAA for four days followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg/l activated charcoal. The rooted plants could be established in soil with more than 90% survival rate.

  2. Effect of gamma radiation on different explants of sweet potato (Ipomoea batatas L. (Lam)) to induce NaCl tolerance

    Energy Technology Data Exchange (ETDEWEB)

    Gutierrez-Rosati, A [Biology Dept., National Agriculture Univ., La Molina, Lima (Peru)

    1997-07-01

    Clonal lines tolerant to NaCl were obtained by combining in vitro culture and gamma radiation in two Peruvian varieties `Amarillo de Quillabamba` and `Nemanete`. The most suitable explants were pedicel sections and leaf blades. Embryogenic callus was induced on basal MS basal medium containing 0.5 ppm 2,4-D. The embryogenic calli were irradiated with 5 Gy from a {sup 137}Cs source. Several putative mutants appeared to be stable. (author). 9 refs, 3 tabs.

  3. Effects of ion beam irradiation on adventitious shoot regeneration from in vitro leaf explants of Septennial ionahta

    International Nuclear Information System (INIS)

    Zhou, L.B.; Li, W.J.; Ma, S.; Dong, X.C.; Yu, L.X.; Li, Q.; Zhou, G.M.; Gao, Q.X.

    2006-01-01

    The effects of 960 MeV carbon ion beam and 8 MeV X-ray irradiation on adventitious shoots from in vitro leaf explants of two different Saintpaulia ionahta (Mauve and Indikon) cultivars were studied with regard to tissue increase, shoots differentiation and morphology changes in the shoots. The experimental results showed that the survival fraction of shoot formation for the Mauve and Indikon irradiated with the carbon ion beam at 20 Gy were 0.715 and 0.600, respectively, while those for both the cultivars exposed to the X-ray irradiation at the same dose were 1.000. Relative biological effectiveness (RBE) of Mauve with respect to X-ray was about two. Secondly, the percentage of regenerating explants with malformed shoots in all Mauve regenerating explants irradiated with carbon ion beam at 20 Gy accounted for 49.6%, while that irradiated with the same dose of X-ray irradiation was only 4.7%; as for Saintpaulia ionahta Indikon irradiated with 20 Gy carbon ion beam, the percentage was 43.3%, which was higher than that of X-ray irradiation. Last, many chlorophyll deficient and other varieties of mutants were obtained in this study. Based on the results above, it can be concluded that the effect of mutation induction by carbon ion beam irradiation on the leaf explants of Saintpaulia ionahta is better than that by X-ray irradiation; and the optimal mutagenic dose varies from 20 Gy to 25 Gy for carbon ion beam irradiation

  4. Relaxin's induction of metalloproteinases is associated with the loss of collagen and glycosaminoglycans in synovial joint fibrocartilaginous explants

    Science.gov (United States)

    Naqvi, Tabassum; Duong, Trang T; Hashem, Gihan; Shiga, Momotoshi; Zhang, Qin; Kapila, Sunil

    2005-01-01

    Diseases of specific fibrocartilaginous joints are especially common in women of reproductive age, suggesting that female hormones contribute to their etiopathogenesis. Previously, we showed that relaxin dose-dependently induces matrix metalloproteinase (MMP) expression in isolated joint fibrocartilaginous cells. Here we determined the effects of relaxin with or without β-estradiol on the modulation of MMPs in joint fibrocartilaginous explants, and assessed the contribution of these proteinases to the loss of collagen and glycosaminoglycan (GAG) in this tissue. Fibrocartilaginous discs from temporomandibular joints of female rabbits were cultured in medium alone or in medium containing relaxin (0.1 ng/ml) or β-estradiol (20 ng/ml) or relaxin plus β-estradiol. Additional experiments were done in the presence of the MMP inhibitor GM6001 or its control analog. After 48 hours of culture, the medium was assayed for MMPs and the discs were analyzed for collagen and GAG concentrations. Relaxin and β-estradiol plus relaxin induced the MMPs collagenase-1 and stromelysin-1 in fibrocartilaginous explants – a finding similar to that which we observed in pubic symphysis fibrocartilage, but not in articular cartilage explants. The induction of these proteinases by relaxin or β-estradiol plus relaxin was accompanied by a loss of GAGs and collagen in joint fibrocartilage. None of the hormone treatments altered the synthesis of GAGs, suggesting that the loss of this matrix molecule probably resulted from increased matrix degradation. Indeed, fibrocartilaginous explants cultured in the presence of GM6001 showed an inhibition of relaxin-induced and β-estradiol plus relaxin-induced collagenase and stromelysin activities to control baseline levels that were accompanied by the maintenance of collagen or GAG content at control levels. These findings show for the first time that relaxin has degradative effects on non-reproductive synovial joint fibrocartilaginous tissue and

  5. Plant regeneration of non-toxic Jatropha curcas—impacts of plant growth regulators, source and type of explants

    KAUST Repository

    Kumar, Nitish; Vijay Anand, K. G.; Reddy, Muppala P.

    2011-01-01

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel plant, however, oil and deoiled cake are toxic. A non-toxic variety of J. curcas is reported from Mexico. The present investigation explores the effects of different plant growth regulators (PGRs) viz. 6-benzyl aminopurine (BAP) or thidiazuron (TDZ) individually and in combination with indole-3-butyric acid (IBA), on regeneration from in vitro and field-grown mature leaf explants, in vitro and glasshouse-grown seedlings cotyledonary leaf explants of non-toxic J. curcas. In all the tested parameters maximum regeneration efficiency (81.07%) and the number of shoot buds per explants (20.17) was observed on 9.08 μM TDZ containing Murashige and Skoog’s (MS) medium from in vitro cotyledonary leaf explants. The regenerated shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with 2.25 μM BAP and 8.5 μM IAA. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing different concentrations and combinations of IBA, IAA and NAA for four days followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg/l activated charcoal. The rooted plants could be established in soil with more than 90% survival rate.

  6. Study on Effect of Type of Explant and Hormone on Callus Induction and Regeneration in Saffron (Crocus sativus L.

    Directory of Open Access Journals (Sweden)

    Mohsen Sajjadi

    2015-10-01

    Full Text Available Saffron (Crocus sativus L. is one of the medicinal plants that contain active components and medicinal materials. Tissue culture of saffron can improve the quality and quantity of the saffron product, increase its export and the farmers’ income. In this study, 36 different types of hormone combinations in the dark and 9 different treatments of hormone combinations in cold (4°C, using different saffron explants (bulb, leaf, scales around leaf and distal parts of the leaf were studied in tissue culture. To investigate the growth of corms, the callus formation and the regeneration rate, three replications for each treatment were used and the length of shoot (cm, the callus formation percentage and the regeneration percentage were measured and statistical analysis was performed. Among the types of explants, only explants from bulbs produced the callus on MS medium containing 2 mg.l-1 BAP and 1 mg.l-1 IBA in both the dark and cold conditions. The highest percentage of regeneration was obtained in MS medium with hormonal composition of 0.3 mg.l-1 TDZ, 1 mg.l-1 BAP, 2 mg.l-1 IBA and 0.01 mg.l-1 GA3 in the cold conditions.

  7. Evaluation of taper joints with combined fatigue and crevice corrosion testing: Comparison to human explanted modular prostheses

    Energy Technology Data Exchange (ETDEWEB)

    Reclaru, L., E-mail: lucien.reclaru@pxgroup.com [PX Group S.A., Dep R and D Corrosion and Biocompatibility Group, Bd. des Eplatures 42, CH-2304 La Chaux-de-Fonds (Switzerland); Brooks, R.A. [Orthopaedic Research, Addenbrooke' s Hospital, University of Cambridge, Box 180 Hills Road, CB2 0QQ Cambridge (United Kingdom); Zuberbühler, M. [Smith and Nephew Orthopaedics AG, Schachenalle 29, 5001 Aarau (Switzerland); Eschler, P.-Y.; Constantin, F. [PX Group S.A., Dep R and D Corrosion and Biocompatibility Group, Bd. des Eplatures 42, CH-2304 La Chaux-de-Fonds (Switzerland); Tomoaia, G. [University of Medicine and Pharmacy Iuliu Hateganu of Cluj-Napoca, Dept. of Orthopaedics and Traumatology, Cluj-Napoca (Romania)

    2014-01-01

    The requirement for revision surgery of total joint replacements is increasing and modular joint replacement implants have been developed to provide adjustable prosthetic revision systems with improved intra-operative flexibility. An electrochemical study of the corrosion resistance of the interface between the distal and proximal modules of a modular prosthesis was performed in combination with a cyclic fatigue test. The complexity resides in the existence of interfaces between the distal part, the proximal part, and the dynamometric screw. A new technique for evaluating the resistance to cyclic dynamic corrosion with crevice stimulation was used and the method is presented. In addition, two components of the proximal module of explanted Ti6Al4V and Ti6Al7Nb prostheses were investigated by optical and electron microscopy. Our results reveal that: The electrolyte penetrates into the interface between the distal and proximal modules during cyclic dynamic fatigue tests, the distal module undergoes cracking and corrosion was generated at the interface between the two models; The comparison of the explanted proximal parts with the similar prostheses evaluated following cyclic dynamic crevice corrosion testing showed that there were significant similarities indicating that this method is suitable for evaluating materials used in the fabrication of modular prostheses. - Highlights: • Electrochemical crevice corrosion testing combined with fatigue test conducted on Ti6Al7Nb and Ti6Al4V modular prostheses • Cations released from integral prostheses • Comparison of human explanted modular prostheses with the similar prostheses evaluated in cyclic dynamic crevice corrosion.

  8. Thidiazuron: A potent cytokinin for efficient plant regeneration in Himalayan poplar (Populus ciliata Wall. using leaf explants

    Directory of Open Access Journals (Sweden)

    Gaurav Aggarwal

    2012-11-01

    Full Text Available Populus species are important resource for certain branches of industry and have special roles for scientific study on biological and agricultural systems. The present investigation was undertaken with an objective of enhancing the frequency of plant regeneration in Himalayan poplar (Populus ciliata Wall.. The effect of Thiadizuron (TDZ alone and in combination with adenine and α-Naphthalene acetic acid (NAA were studied on the regeneration potential of leaf explants. A high efficiency of shoot regeneration was observed in leaf (80.00% explants on MS basal medium supplemented with 0.024 mg/l TDZ and 79.7 mg/l adenine. Elongation and multiplication of shoots were obtained on Murashige and Skoog (MS basal medium, containing 0.5 mg/l 6. Benzyl aminopurine (BAP + 0.2mg/l Indole 3-acetic acid (IAA + 0.3 mg/l Gibberellic acid (GA3. High frequency root regeneration from in vitro developed shoots was observed on MS basal medium supplemented with 0.10 mg/l Indole 3-butyric acid(IBA. Maximum of the in vitro rooted plantlets were well accomplished to the mixture of sand: soil (1:1 and exhibited similar morphology with the field plants. A high efficiency plant regeneration protocol has been developedfrom leaf explants in Himalayan poplar (Populus ciliata Wall..

  9. Effect of ZnO nanoparticles on Brassica nigra seedlings and stem explants: growth dynamics and antioxidative response

    Directory of Open Access Journals (Sweden)

    Hira eZafar

    2016-04-01

    Full Text Available Nanoparticles (NPs have diverse properties in comparison to respective chemicals due to structure, surface area ratio, morphology, and reactivity. Toxicological effects of metallic NPs to organisms including plants have been reported. However, to the authors’ knowledge there is no report on the effect of NPs on in vitro culture of plant explants. In this study, ZnO NPs at 500-1500 mg/L badly affected Brassica nigra seed germination and seedling growth and raised antioxidative activities and antioxidants concentrations. On the other hand, culturing the stem explants of B. nigra on Murashige and Skoog (MS medium in presence of low concentration of ZnO NPs (1-20 mg/L produced white thin roots with thick root hairs. At 10 mg/L ZnO NPs shoots emergence was also observed. The developed calli/roots showed 79% DPPH (2,2-diphenyl-1-picryl hydrazyl radical scavenging activity at 10 mg/L. While total antioxidant and reducing power potential were also significantly different in presence of ZnO NPs. Non enzymatic antioxidative molecules, phenolics (up to 0.15 µg GAE/mg FW and flavonoids (up to 0.22 µg QE/mg FW, also raised and found NPs concentration dependent. We state that ZnO NPs may induce roots from explants cultured on appropriate medium and can be cultured for production of valuable secondary metabolites.

  10. Refining the application of direct embryogenesis in sugarcane: Effect of the developmental phase of leaf disc explants and the timing of DNA transfer on transformation efficiency.

    Science.gov (United States)

    Snyman, S J; Meyer, G M; Richards, J M; Haricharan, N; Ramgareeb, S; Huckett, B I

    2006-10-01

    A rapid in vitro protocol using direct somatic embryogenesis and microprojectile bombardment was investigated to establish the developmental phases most suitable for efficient sugarcane transformation. Immature leaf roll disc explants with and without pre-emergent inflorescence tissue were compared. It was shown that for effective transformation to occur, explants should be cultured for several days to allow initiation of embryo development prior to bombardment. Leaf roll discs with pre-emergent inflorescences showed a higher degree of embryogenic competence than non-flowering explants, and transformation efficiency was higher when explants containing floral initials were bombarded. Despite the occurrence of high numbers of phenotypically negative plants, combining the use of inflorescent leaf roll discs with direct embryogenic regeneration has the potential to improve the speed and efficiency of transgenesis in sugarcane.

  11. Status of the segment interconnect, cable segment ancillary logic, and the cable segment hybrid driver projects

    International Nuclear Information System (INIS)

    Swoboda, C.; Barsotti, E.; Chappa, S.; Downing, R.; Goeransson, G.; Lensy, D.; Moore, G.; Rotolo, C.; Urish, J.

    1985-01-01

    The FASTBUS Segment Interconnect (SI) provides a communication path between two otherwise independent, asynchronous bus segments. In particular, the Segment Interconnect links a backplane crate segment to a cable segment. All standard FASTBUS address and data transactions can be passed through the SI or any number of SIs and segments in a path. Thus systems of arbitrary connection complexity can be formed, allowing simultaneous independent processing, yet still permitting devices associated with one segment to be accessed from others. The model S1 Segment Interconnect and the Cable Segment Ancillary Logic covered in this report comply with all the mandatory features stated in the FASTBUS specification document DOE/ER-0189. A block diagram of the SI is shown

  12. Effect Of Explant Source And Different Medium Culture On Friable Embryogenic Callus Induction Of Four Cultivars Of Cassava Manihot Esculenta Crantz

    Directory of Open Access Journals (Sweden)

    Simplice Prosper Yandia

    2015-08-01

    Full Text Available In order to obtain Friable Embryogenic Callus FEC for protoplast isolation we have evaluated in this research the competance for Friable Embryogenic Callus FEC of four cassava cultivars M61033 Rendre Yalipe and Six-mois in media containing MS supplemented with 8mgl 24-D MS supplemented with 10 mgl BAP and GD supplemented with 12mgl picloram using apical bud AB and immature leaves lobes ILL as explants. In general in the medium GD12mgl picloram the highest efficiencies of FEC ranged from 58 to 87 and the highest score of FEC ranged from 4.2 to 5.4 with explants AB however we have observed with explants ILL the efficiencies of somatic embryos ranged form 41 to 75 and the score ranged from 4.1 to 4.4. The mediums MS28 mgl 24-D have induced with explants AB the efficiencies of FEC ranged from 43 to 57 and the score ranged from 3.1 to 3.8 however with ILL explants the efficiencies of FEC ranged from 39 to 49 and the score ranged from 2.9 to 3.7. The least FEC were observed in the medium MS210 mgl BAP with BA explants however the efficiencies ranged from 6 to 11 and the score ranged from 1.1 to 1.8. Whereas the efficiencies of FEC with ILL explants ranged from 4 to 7 and the score ranged from 0.5 to 0.8. All of four cultivars showed capability of producing FEC although their efficiency varied according to gonotype donors explants and medium taking into acount. Abbreviations GD Gressoff and Doy MS Murashige and Skoog 24-dichloro phenoxyacetic acid BAP Benzylamino-purin-Acid AB Apical Bud ILL Immature Leaves lobes

  13. In vtro adventitious shoot regeneration from cotyledon explant of brassica oleracea subsp. Italica and brassica oleracea subsp. capitata using tdz and naa

    International Nuclear Information System (INIS)

    Salim, S.; Rashid, A.

    2014-01-01

    Broccoli(Brassica oleracea subsp. italica) cv. Green Dragon King and cabbage (Brassica oleracea subsp. capitata) cv. Gianty are important vegetable crops grown in Cameron Highlands, Malaysia. The cotyledons of both cultivars were used as explant source for in vitro shoot regeneration. The objective of this research was to examine the influence of the growth regulators thidiazuron (TDZ) and naphthaleneacetic acid (NAA) on adventitious shoot formation in these cultivars. This system of adventitious shoot regeneration from cotyledon explants could be useful as a tool for genetic transformation of the subspecies. Cotyledon explants of both cultivars excised from 5-day-old in vitro germinated seedlings were placed on shoot induction medium containing basal salts of Murashige and Skoog (MS) and various concentrations of TDZ and NAA. The highest percentage of cotyledon explant of broccoli cv. Green Dragon King producing shoot (76.66%) and the highest mean number of shoots produced per explant (0.9) were obtained on 0.1 mg/l TDZ with 0.1 mg/l NAA. Meanwhile, the highest percentage of cotyledon explant of cabbage cv. Gianty producing shoots (86.67%) and highest number of shoots produced per explant (1.1) were recorded on 0.5 mg/l TDZ with 0.1 mg/l NAA. Therefore, 0.1 mg/l TDZ with 0.1 mg/l NAA and 0.5 mg/l TDZ with 0.1mg/l NAA are the recommended combinations for adventitious shoot regeneration from cotyledonary explants of broccoli cv. Dragon King and cabbage cv. Gianty respectively. (author)

  14. Market segmentation: Venezuelan ADRs

    Directory of Open Access Journals (Sweden)

    Urbi Garay

    2012-12-01

    Full Text Available The control on foreign exchange imposed by Venezuela in 2003 constitute a natural experiment that allows researchers to observe the effects of exchange controls on stock market segmentation. This paper provides empirical evidence that although the Venezuelan capital market as a whole was highly segmented before the controls were imposed, the shares in the firm CANTV were, through their American Depositary Receipts (ADRs, partially integrated with the global market. Following the imposition of the exchange controls this integration was lost. Research also documents the spectacular and apparently contradictory rise experienced by the Caracas Stock Exchange during the serious economic crisis of 2003. It is argued that, as it happened in Argentina in 2002, the rise in share prices occurred because the depreciation of the Bolívar in the parallel currency market increased the local price of the stocks that had associated ADRs, which were negotiated in dollars.

  15. Scintillation counter, segmented shield

    International Nuclear Information System (INIS)

    Olson, R.E.; Thumim, A.D.

    1975-01-01

    A scintillation counter, particularly for counting gamma ray photons, includes a massive lead radiation shield surrounding a sample-receiving zone. The shield is disassembleable into a plurality of segments to allow facile installation and removal of a photomultiplier tube assembly, the segments being so constructed as to prevent straight-line access of external radiation through the shield into radiation-responsive areas. Provisions are made for accurately aligning the photomultiplier tube with respect to one or more sample-transmitting bores extending through the shield to the sample receiving zone. A sample elevator, used in transporting samples into the zone, is designed to provide a maximum gamma-receiving aspect to maximize the gamma detecting efficiency. (U.S.)

  16. Head segmentation in vertebrates

    OpenAIRE

    Kuratani, Shigeru; Schilling, Thomas

    2008-01-01

    Classic theories of vertebrate head segmentation clearly exemplify the idealistic nature of comparative embryology prior to the 20th century. Comparative embryology aimed at recognizing the basic, primary structure that is shared by all vertebrates, either as an archetype or an ancestral developmental pattern. Modern evolutionary developmental (Evo-Devo) studies are also based on comparison, and therefore have a tendency to reduce complex embryonic anatomy into overly simplified patterns. Her...

  17. Growth of vegetative explant Moringa oleifera on different composition of auxin and cytokinin and its synthetic seed germination

    Science.gov (United States)

    Muslihatin, Wirdhatul; Jadid, Nurul; Puspitasari, Ika D.; Safitri, Chusnul E.

    2017-06-01

    The spread of Moringa oleifera is also rare for seed germination and viability or survival are low, and the lack of vegetative propagation method. The purpose of this study are to determine the effect of auxin and cytokinin on growth vegetative explants Moringa oleifera and its synthetic seed germination. The explants grown on MS medium with sucrose content of 30% and a range of additional hormone. Addition concentration and different types of hormone made in order to know the sensitivity and response explant growth on a variety of media to get a good callus and embryosomatic. The composition of the hormone given is MS + 2.4 D 3 ppm; MS + 2,4D 2 ppm + BAP 2 ppm; MS + NAA + 0.5 ppm kinetin 1 ppm; MS + NAA 1 ppm + kinetin 1 ppm; MS + NAA 1 ppm + 0.5 ppm kinetin. The explants were incubated at a temperature of 18-20 ° C with a photoperiod 16/8. Explants and MS medium is incubated to form embryonic callus. Seeds synthetic made from embryonic callus growing on medium 1 ppm kinetin + NAA 1 ppm with encapsulation method with sodium alginate 2%. Seed synthetic germinated in some kind of medium that medium ms0 solid (M1), ms0 liquid (M2), MS0 semi-solid (M3), MS solid NAA 1ppm + Kinetin 1 ppm (M4), MS liquid NAA 1 ppm + kinetin (M5), and semi-solid MS + NAA 1 ppm kinetin 1 ppm (M6). Synthetic seed viability was observed with the parameters of the fresh weight of synthetic seed, germination percentage and seedling. Chlorophyll content was measured by spectrophotometric method with solvent asseton. Best callus generated in this study are embryonic callus that grew on media NAA 1 ppm + kinetin 1 ppm. Embryonic callus on M6 + NAA 1 ppm kinetin 1 ppm capable of germination with an average weight of callus and sprouts of 40.38 mg. Of the entire amount of a synthetic seed on M6, just 5 seed germinate, so the percentage of germination of seeds is equal to 41.67%. with an average length of sprouts 1 cm with an average total chlorophyll content of 8.66 mg / g.

  18. Sacha Inchi Oil (Plukenetia volubilis L.), effect on adherence of Staphylococus aureus to human skin explant and keratinocytes in vitro.

    Science.gov (United States)

    Gonzalez-Aspajo, German; Belkhelfa, Haouaria; Haddioui-Hbabi, Laïla; Bourdy, Geneviève; Deharo, Eric

    2015-08-02

    Plukenetia volubilis L. (Euphorbiaceae) is a domesticated vine distributed from the high-altitude Andean rain forest to the lowlands of the Peruvian Amazon. Oil from the cold-pressed seeds, sold under the commercial name of Sacha Inchi Oil (SIO) is actually much in favour because it contains a high percentage of omega 3 and omega 6, and is hence used as a dietary supplement. SIO is also used traditionally for skin care, in order to maintain skin softness, and for the treatment of wounds, insect bites and skin infections, in a tropical context where the skin is frequently damaged. This study was designed in order to verify whether the traditional use of SIO for skin care would have any impact on Staphylococcus aureus growth and skin adherence, as S. aureus is involved in many skin pathologies (impetigo, folliculitis, furuncles and subcutaneous abscesses) being one if the main pathogens that can be found on the skin. Therefore, our objective was to assess SIO bactericidal activity and interference with adherence to human skin explants and the keratinocyte cell line. Cytotoxicity on that cells was also determined. The activity of SIO was compared to coconut oil (CocO), which is widely used for skin care but has different unsaturated fatty acids contents. Laboratory testing with certified oil, determined antibacterial activity against radio labelled S. aureus. Cytotoxic effects were measured with XTT on keratinocyte cells and with neutral red on human skin explants; phenol was used as cytotoxic control. Adherence assays were carried out by mixing H3-labelled S. aureus bacteria with keratinocyte cells and human skin explants, incubated with oils 2h before (to determine the inhibition of adherence, assimilated to a preventive effect) or 2h after the contact of the biological material with S. aureus (to assess the detachment of the bacteria, assimilated to a curative effect). Residual radioactivity measured after washings made it possible to determine the adherence

  19. Replication of avian, human and swine influenza viruses in porcine respiratory explants and association with sialic acid distribution

    Directory of Open Access Journals (Sweden)

    Nauwynck Hans J

    2010-02-01

    Full Text Available Abstract Background Throughout the history of human influenza pandemics, pigs have been considered the most likely "mixing vessel" for reassortment between human and avian influenza viruses (AIVs. However, the replication efficiencies of influenza viruses from various hosts, as well as the expression of sialic acid (Sia receptor variants in the entire porcine respiratory tract have never been studied in detail. Therefore, we established porcine nasal, tracheal, bronchial and lung explants, which cover the entire porcine respiratory tract with maximal similarity to the in vivo situation. Subsequently, we assessed virus yields of three porcine, two human and six AIVs in these explants. Since our results on virus replication were in disagreement with the previously reported presence of putative avian virus receptors in the trachea, we additionally studied the distribution of sialic acid receptors by means of lectin histochemistry. Human (Siaα2-6Gal and avian virus receptors (Siaα2-3Gal were identified with Sambucus Nigra and Maackia amurensis lectins respectively. Results Compared to swine and human influenza viruses, replication of the AIVs was limited in all cultures but most strikingly in nasal and tracheal explants. Results of virus titrations were confirmed by quantification of infected cells using immunohistochemistry. By lectin histochemistry we found moderate to abundant expression of the human-like virus receptors in all explant systems but minimal binding of the lectins that identify avian-like receptors, especially in the nasal, tracheal and bronchial epithelium. Conclusions The species barrier that restricts the transmission of influenza viruses from one host to another remains preserved in our porcine respiratory explants. Therefore this system offers a valuable alternative to study virus and/or host properties required for adaptation or reassortment of influenza viruses. Our results indicate that, based on the expression of Sia

  20. Video segmentation using keywords

    Science.gov (United States)

    Ton-That, Vinh; Vong, Chi-Tai; Nguyen-Dao, Xuan-Truong; Tran, Minh-Triet

    2018-04-01

    At DAVIS-2016 Challenge, many state-of-art video segmentation methods achieve potential results, but they still much depend on annotated frames to distinguish between background and foreground. It takes a lot of time and efforts to create these frames exactly. In this paper, we introduce a method to segment objects from video based on keywords given by user. First, we use a real-time object detection system - YOLOv2 to identify regions containing objects that have labels match with the given keywords in the first frame. Then, for each region identified from the previous step, we use Pyramid Scene Parsing Network to assign each pixel as foreground or background. These frames can be used as input frames for Object Flow algorithm to perform segmentation on entire video. We conduct experiments on a subset of DAVIS-2016 dataset in half the size of its original size, which shows that our method can handle many popular classes in PASCAL VOC 2012 dataset with acceptable accuracy, about 75.03%. We suggest widely testing by combining other methods to improve this result in the future.

  1. Market segmentation in behavioral perspective.

    OpenAIRE

    Wells, V.K.; Chang, S.W.; Oliveira-Castro, J.M.; Pallister, J.

    2010-01-01

    A segmentation approach is presented using both traditional demographic segmentation bases (age, social class/occupation, and working status) and a segmentation by benefits sought. The benefits sought in this case are utilitarian and informational reinforcement, variables developed from the Behavioral Perspective Model (BPM). Using data from 1,847 consumers and from a total of 76,682 individual purchases, brand choice and price and reinforcement responsiveness were assessed for each segment a...

  2. Early somatic embryogenesis in Heliconia chartacea Lane ex Barreiros cv. Sexy Pink ovary section explants

    Directory of Open Access Journals (Sweden)

    Cláudia Ulisses

    2010-02-01

    Full Text Available The present work evaluated the development of embryogenic callus from transversal ovary sections. The experiments were carried out under two experimental regimes using combinations of IAA (0; 5.71; 8.56; 11.42; 14.27μM and 2,4-D (0; 13.57; 18.10; 22.62μM or combinations of 2,4-D with BA (0; 4.43; 6.65; 8.87; 11.09μM. Assessments were made of anatomical aspects of the callus and for the presence of embryogenic structures using cytochemical and histological analyses and stereomicroscopic and scanning electronic microscopic observations. Treatments with 2,4-D and IAA produced friable calluses demonstrating cellular acquisition of morphogenetic competence as well as the formation of pro-embryogenic sectors. The expression of embryogenic program could be observed, with proembryogenic cell clusters developing into globular embryos. These results offer the possibility of using new types of explants for culturing helicons that avoid the growth of endophytic bacteria.Este trabalho teve como objetivo avaliar a resposta de secções transversais de ovários e o desenvolvimento de calos embriogênicos. O experimento constou de dois ensaios. No primeiro avaliou-se combinações entre AIA (0; 5.71; 8.56; 11.42; 14.27μM e 2,4-D (0; 13.57; 18.10; 22.62μM e no segundo avaliou-se as concentrações de 2,4-D supracitadas, combinadas com concentrações de BA (0; 4.43; 6.65; 8.87; 11.09μM. Os calos formados foram avaliados quanto à presença de estruturas embriogênicas utilizando-se estereomicroscópio, microscópio eletrônico de varredura, além de análises citoquímicas e histológicas. Combinações entre 2,4-D e AIA induziram a formação de calos friáveis com setores pró-embriogênicos, refletindo a aquisição de competência morfogenética. Posteriormente foi observada a expressão do programa embriogênico quando massas pró-embriogências desenvolveram-se formando embriões somáticos. Esses resultados apresentam uma alternativa para a utiliza

  3. MIV-150-containing intravaginal rings protect macaque vaginal explants against SHIV-RT infection.

    Science.gov (United States)

    Ouattara, Louise A; Barnable, Patrick; Mawson, Paul; Seidor, Samantha; Zydowsky, Thomas M; Kizima, Larisa; Rodriguez, Aixa; Fernández-Romero, José A; Cooney, Michael L; Roberts, Kevin D; Gettie, Agegnehu; Blanchard, James; Robbiani, Melissa; Teleshova, Natalia

    2014-05-01

    Recent studies demonstrated that intravaginal rings (IVRs) containing 100 mg of the nonnucleoside reverse transcriptase inhibitor (NNRTI) MIV-150 significantly protect macaques against a chimeric simian-human immunodeficiency virus that expresses the HIV-1 HxB2 reverse transcriptase (SHIV-RT) when present before and after vaginal challenge. The objectives of this study were to (i) evaluate the pharmacodynamics (PD) of MIV-150 in vaginal fluids (VF) and in ectocervical and vaginal tissues following 100-mg MIV-150 IVR exposure and to (ii) gain more insight whether pharmacokinetics (PK) of MIV-150 can predict PD. MIV-150 in VF collected at 1 day and 14 days post-MIV-150 IVR insertion inhibited ex vivo SHIV-RT infection in vaginal biopsy specimens from untreated animals (not carrying IVRs) in a dose-dependent manner. Previous PK studies demonstrated a significant increase of ectocervical and vaginal tissue MIV-150 concentrations 14 days versus 1 day post-IVR insertion, with the highest increase in vaginal tissue. Therefore, we tested PD of MIV-150 in tissues 14 days post-MIV-150 IVR insertion. Ex vivo SHIV-RT infection of vaginal, but not ectocervical, tissues collected 14 days post-MIV-150 IVR insertion was significantly inhibited compared to infection at the baseline (prior to MIV-150 IVR exposure). No changes in vaginal and ectocervical tissue infection were observed after placebo IVR exposure. Overall, these data underscore the use of the ex vivo macaque explant challenge models to evaluate tissue and VF PK/PD of candidate microbicides before in vivo animal efficacy studies. The data support further development of MIV-150-containing IVRs.

  4. The diagnosis of breast implant rupture: MRI findings compared with findings at explantation

    International Nuclear Information System (INIS)

    Hoelmich, Lisbet R.; Vejborg, Ilse; Conrad, Carsten; Sletting, Susanne; McLaughlin, Joseph K.

    2005-01-01

    Study objective: The aim of this study was to evaluate the accuracy of Magnetic Resonance Imaging (MRI) as performed according to a strict study protocol in diagnosing rupture of silicone breast implants. Material and methods: The study population consisted of 64 women with 118 implants, who had participated in either one or two study MRI examinations, aiming at determining the prevalence and incidence of silent implant rupture, respectively, and who subsequently underwent explantation. Implant rupture status was determined by four independent readers and a consensus diagnosis of either rupture (intracapsular or extracapsular), possible rupture or intact implant was then obtained. Strict predetermined rupture criteria were applied as described in this report and findings at surgery were abstracted in a standardised manner and results compared. Results: At MRI, 66 implants were diagnosed as ruptured, nine as possibly ruptured and 43 as intact. Among the ruptured implants, 27 were categorized as extracapsular. At surgery, on average 297 days after the MRI, 65 of the 66 rupture diagnoses were confirmed, as were 20 of the cases with extracapsular silicone. Eight of the nine possibly ruptured implants were in fact ruptured at surgery. Thirty-four of the 43 intact implants were described as intact at surgery. When categorising possible ruptures as ruptures, there were one false positive and nine false negative rupture diagnoses at MRI yielding an accuracy of 92%, a sensitivity of 89%, and a specificity of 97%. Correspondingly, the predictive value of a positive MRI examination was 99% and the predictive value of a negative MRI examination was 79%. Conclusions: We conclude that MRI is highly accurate for identification of silicone breast implant rupture, with a high sensitivity and specificity when evaluation of images are based on presence of well-defined rupture criteria

  5. Shoot regeneration from cotyledonary leaf explants of jatropha curcas: A biodiesel plant

    KAUST Repository

    Kumar, Nitish Chandramohana

    2010-03-07

    A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds were transferred to MS medium containing 10 lM kinetin (Kn), 4.5 lM BAP, and 5.5 lM a-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 lM BAP and 8.5 lM IAA was found to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing dif- ferent concentrations and combinations of IBA, IAA, and NAA for 4 days, followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l-1 activated charcoal. Elongated shoot treated with 15 lM IBA, 5.7 lM IAA, and 11 lM NAA resulted in highest percent rooting. The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement of J. curcas through genetic modification. © Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2010.

  6. Intestinal toxicity of deoxynivalenol is limited by Lactobacillus rhamnosus RC007 in pig jejunum explants.

    Science.gov (United States)

    García, Gisela Romina; Payros, Delphine; Pinton, Philippe; Dogi, Cecilia Ana; Laffitte, Joëlle; Neves, Manon; González Pereyra, María Laura; Cavaglieri, Lilia Renée; Oswald, Isabelle P

    2018-02-01

    Probiotics have been explored to stimulate gut health in weaned pigs, when they started to consume solid diet where mycotoxins could be present. The aim of this study was to evaluate the effect of Lactobacillus rhamnosus RC007 on the intestinal toxicity of deoxynivalenol (DON) in an ex vivo model. Jejunal explants, obtained from 5-week-old crossbred castrated male piglets, were kept as control, exposed for 3 h to 10 μM DON, incubated for 4 h with 10 9 CFU/mL L. rhamnosus, or pre-incubated 1 h with 10 9 L. rhamnosus and exposed to DON. Histological lesions were observed, para- and transcellular intestinal permeability was measured in Ussing chambers. The expression levels of mRNA encoding six inflammatory cytokines (CCL20, IL-10, IL-1β, TNFα, IL-8 and IL-22) were determined by RT-PCR. The expressions of the phosphorylated MAP kinases p42/p44 and p38 were assessed by immunoblotting. Exposure to DON induced histological changes, significantly increased the expression of CCL20, IL-1β, TNFα, IL-8, IL-22 and IL-10, increased the intestinal paracellular permeability and activated MAP kinases. Incubation with L. rhamnosus alone did not have any significant effect. By contrast, the pre-incubation with L. rhamnosus reduced all the effects of DON: the histological alterations, the pro-inflammatory response, the paracellular permeability and the phosphorylation of MAP kinases. Of note, L. rhamnosus did not adsorb DON and only slightly degrade the toxin. In conclusion, L. rhamnosus RC007 is a promising probiotic which, included as feed additive, can decrease the intestinal toxicity of DON.

  7. Shoot regeneration from cotyledonary leaf explants of jatropha curcas: A biodiesel plant

    KAUST Repository

    Kumar, Nitish Chandramohana; Vijay Anand, K. G.; Reddy, Muppala P.

    2010-01-01

    A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds were transferred to MS medium containing 10 lM kinetin (Kn), 4.5 lM BAP, and 5.5 lM a-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 lM BAP and 8.5 lM IAA was found to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing dif- ferent concentrations and combinations of IBA, IAA, and NAA for 4 days, followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l-1 activated charcoal. Elongated shoot treated with 15 lM IBA, 5.7 lM IAA, and 11 lM NAA resulted in highest percent rooting. The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement of J. curcas through genetic modification. © Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2010.

  8. Indirect organogenesis in milkweed ( Calotropis procera from mature zygotic embryo explants

    Directory of Open Access Journals (Sweden)

    Hojatollah Abbasi

    2018-01-01

    Full Text Available Milkweed ( Calotropis procera is a valuable medicinal plant which grows in many regions of Iran. Its significant medicinal properties have made it an important crop which is cultivated commercially. This plant is propagated from seeds as well as root and shoot cuttings. Due to problems in the usage of these reproduction methods, new propagation methods such as tissue culturing should be developed. This study was aimed at obtaining appropriate concentrations of plant hormones for indirect organogenesis of milkweed. The experiment was arranged in a completely randomized design (CRD with 3 replications. The effects of various concentrations of (2,4-dichlorophenoxyacetic acid 2,4-D (0.1, 0.5, 1, 2 and 3 mg/l were studied in terms of callus induction and shoot regeneration on an MS based medium supplemented with BA (benzyl amino purine and NAA (naphthalene acetic acid at the same concentration. Mature embryos were used as explants and morphological traits such as embryo size, callus size, number and size of shoots and roots were recorded. The results showed that 2,4-D significantly increased the size of cultured embryos (P < 0.05. The largest embryo volume was observed in cultures treated with 3 mg/l 2,4-D. The highest callusing was recorded in 2 mg/l 2,4-D. The effects of BA and NAA concentrations on shoot regeneration were significant and the highest values were observed for a combination of 1 mg/l BA and 2 mg/l NAA. 1 mg/l IBA (Indole 3-butyric acid was able to induce the highest number of better quality roots and shoots.

  9. Segmenting the Adult Education Market.

    Science.gov (United States)

    Aurand, Tim

    1994-01-01

    Describes market segmentation and how the principles of segmentation can be applied to the adult education market. Indicates that applying segmentation techniques to adult education programs results in programs that are educationally and financially satisfying and serve an appropriate population. (JOW)

  10. Market Segmentation for Information Services.

    Science.gov (United States)

    Halperin, Michael

    1981-01-01

    Discusses the advantages and limitations of market segmentation as strategy for the marketing of information services made available by nonprofit organizations, particularly libraries. Market segmentation is defined, a market grid for libraries is described, and the segmentation of information services is outlined. A 16-item reference list is…

  11. Effect of 6-BA on nodal explant bud sproutings of Coffea arabica cv. Mundo Novo Efeito de 6-BA na brotação de gemas de explantes nodais de Coffea arabica cv. Mundo Novo

    Directory of Open Access Journals (Sweden)

    Luis Carlos da Silva Ramos

    2005-01-01

    Full Text Available Coffee plants can be micropropagated by nodal bud sprouting using the 6-benzylaminopurine (6-BA hormone. However, literature reports the use of a wide range of 6-BA, from 0.5 to 88.8 µM L-1. So, this study was performed to narrow that range. Nodal explants of Coffea arabica cv Mundo Novo obtained from in vitro plantlets were inoculated on gelled-MS medium supplemented with different concentrations of 6-BA. Two assays were carried out: in the first one, 6-BA was used at concentrations of 0, 5, 25, 50, and 100 µM L-1, being evaluated at 43 and 123 days. In the second experiment, dosis of 10, 20 and 30 µM L-1, have evaluated at 65 and 100 days. Treatments with 6-BA induced multiple sprouting from the nodal explants, which were best characterized around 100 days after inoculation. The nodal explants grew taller and showed multiple shoots, whereas the effect of 6-BA at 5 to 25 µM L-1 was similar to that with higher concentrations (50 and 100 µM L-1. Nodal explants yielded from 2.9 to 6.0 buds per node, achieving height of 1.3 to 1.5 cm at 5 to 25 µM L-1 of 6-BA, whereas they yielded from 4.3 to 4.9 buds per node but the sprouting grew about 0.8 cm at 50 and 100 µM L-1 of 6-BA. This study indicated that multiple sprouting of lateral buds can be induced by lower concentrations of 6-BA, for example, from 10 to 30 µM L-1, diminishing possible risks of somaclonal variation due to high levels of hormone concentration.O cafeeiro pode ser micropropagado via brotação de gemas laterais, aplicando o regulador de crescimento 6-benzilaminopurina (6-BA. Entretanto, a literatura apresenta ampla variação da dose empregada, desde 0.5 a 88.8 µM L-1. Assim, este estudo visou otimizar doses para explantes nodais do cafeeiro C. arabica cv Mundo Novo. Explantes nodais, obtidos de plântulas cultivadas in vitro, foram inoculados em meio MS geleificado, com adição de diferentes concentrações de 6-BA. Foram feitos dois experimentos: no primeiro, 6-BA foi

  12. Albedo estimation for scene segmentation

    Energy Technology Data Exchange (ETDEWEB)

    Lee, C H; Rosenfeld, A

    1983-03-01

    Standard methods of image segmentation do not take into account the three-dimensional nature of the underlying scene. For example, histogram-based segmentation tacitly assumes that the image intensity is piecewise constant, and this is not true when the scene contains curved surfaces. This paper introduces a method of taking 3d information into account in the segmentation process. The image intensities are adjusted to compensate for the effects of estimated surface orientation; the adjusted intensities can be regarded as reflectivity estimates. When histogram-based segmentation is applied to these new values, the image is segmented into parts corresponding to surfaces of constant reflectivity in the scene. 7 references.

  13. Indução de calos friáveis em explantes foliares de Salix (Salyx humboldtiana Willd Induction of friable callus in leaf explants of Salix (Salyx humboldtiana Willd

    Directory of Open Access Journals (Sweden)

    Breno Régis Santos

    2005-06-01

    Full Text Available O salix é uma árvore ornamental adaptada a terrenos úmidos, sendo considerada uma espécie importante para a recomposição de áreas ciliares degradadas. A madeira pode ser empregada na indústria em geral. Apesar de produzir grande quantidade de sementes, estas não possuem alta percentagem de germinação. O presente trabalho teve como objetivo realizar o estabelecimento in vitro de salix através da indução e formação de calos friáveis em explantes foliares, visando a uma posterior regeneração de plantas para a propagação massal desta espécie. Explantes foliares foram inoculados em meio de cultura MS (MURASHIGE & SKOOG, 1962, acrescido de diferentes concentrações de ácido 2,4-diclorofenoxiacético (0; 1,0; 2,0; 4,0; 6,0; 8,0; 10,0 e 12,0mg L-1 e combinações entre ácido naftalenoacético e benzilaminopurina, ambos em concentrações de 0,0; 1,0; 2,0; 4,0; 6,0 e 8,0mg L-1. Os resultados demonstram que explantes inoculados na ausência de reguladores de crescimento não apresentam a formação de calos friáveis. Significativa produção de calos friáveis (90% é obtida utilizando-se 6,0mg L-1 de ácido 2,4-diclorofenoxiacético. A utilização de concentração individual de ácido naftalenoacético ou benzilaminopurina, além de induzir a calogênese também foi capaz de promover rizogênese.Salix is an ornamental tree adapted to humid soils being considered an important species used in depleted areas. Its wood may be used in all kinds of industries. Although the production of seeds is large, the germination is reduced. The objective of this work was to establish salix in vitro through the induction and formation of friable callus of leaf explants to provide future regeneration of plants for mass propagation of the species. Leaf explants were inoculated in MS medium (MURASHIGE & SKOOG, 1962 supplemented with different concentrations of 2-4-diclorofenoxiacetic acid (0; 1.0; 2.0; 4.0; 6.0; 8.0 and 12.0mg L-1 and combinations

  14. FRAMEWORK FOR COMPARING SEGMENTATION ALGORITHMS

    Directory of Open Access Journals (Sweden)

    G. Sithole

    2015-05-01

    Full Text Available The notion of a ‘Best’ segmentation does not exist. A segmentation algorithm is chosen based on the features it yields, the properties of the segments (point sets it generates, and the complexity of its algorithm. The segmentation is then assessed based on a variety of metrics such as homogeneity, heterogeneity, fragmentation, etc. Even after an algorithm is chosen its performance is still uncertain because the landscape/scenarios represented in a point cloud have a strong influence on the eventual segmentation. Thus selecting an appropriate segmentation algorithm is a process of trial and error. Automating the selection of segmentation algorithms and their parameters first requires methods to evaluate segmentations. Three common approaches for evaluating segmentation algorithms are ‘goodness methods’, ‘discrepancy methods’ and ‘benchmarks’. Benchmarks are considered the most comprehensive method of evaluation. This paper shortcomings in current benchmark methods are identified and a framework is proposed that permits both a visual and numerical evaluation of segmentations for different algorithms, algorithm parameters and evaluation metrics. The concept of the framework is demonstrated on a real point cloud. Current results are promising and suggest that it can be used to predict the performance of segmentation algorithms.

  15. Agrobacterium rhizogenes-dependent production of transformed roots from foliar explants of pepper (Capsicum annuum): a new and efficient tool for functional analysis of genes

    OpenAIRE

    Aarrouf, Jawad; Mallard, Stephanie; Caromel, Bernard; Lizzi, Y.; Lefebvre, Véronique

    2012-01-01

    Pepper is known to be a recalcitrant species to genetic transformation via Agrobacterium tumefaciens. A. rhizogenes-mediated transformation offers an alternative and rapid possibility to study gene functions in roots. In our study, we developed a new and efficient system for A. rhizogenes transformation of the cultivated species Capsicum annuum. Hypocotyls and foliar organs (true leaves and cotyledons) of Yolo Wonder (YW) and Criollo de Morelos 334 (CM334) pepper cultivars were inoculated wit...

  16. The Pattern of Distribution of Peroxidase and H{sub 2}O{sub 2} in Hypocotyls of Pumpkin irradiated with Gamma Ray

    Energy Technology Data Exchange (ETDEWEB)

    Wi, Seung Gon; Chung, Byung Yeoup; Kim, Jae Sung; Kim, Jin Hong; Lee, Ju Woon; Baek, Myung Hwa; Chae, Hyo Seok [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

    2005-07-01

    Gamma radiation, one of ionizing radiations, has been reported to affect the morphological, anatomical, biochemical and physiological changes of plants at different dose levels. These effects at high level include inhibition in plant growth. Peroxidases (PODs) are mainly participations the process of lignification on the cell wall and protect the cell organelles in cytosol against the oxidative stresses by ROS (reactive oxygen species). Hydrogen peroxide (ROS) is normal metabolite in aerobic cells and the physiological steady concentrations (between 10{sup -7} and 10{sup -9}) are not particularly cytotoxic. When these concentrations are increased by ionizing radiation, they lead to cell lethality. Thus radiation-induced H{sub 2}O{sub 2} may appear as an important agent causing cell damage. In this study, a polyclonal antibody against peroxidase and cerium chloride as a trapping agent for H{sub 2}O{sub 2} were used to obtain for better information on the occurrence and distribution of POD and H{sub 2}O{sub 2} in the cytoplasm and walls of vascular bundle in hypocotyls of pumpkin.

  17. Association between IL-6 production in synovial explants from rheumatoid arthritis patients and clinical and imaging response to biologic treatment: A pilot study.

    Directory of Open Access Journals (Sweden)

    Martin Andersen

    Full Text Available The need for biomarkers which can predict disease course and treatment response in rheumatoid arthritis (RA is evident. We explored whether clinical and imaging responses to biologic disease modifying anti-rheumatic drug treatment (bDMARD were associated with the individual's mediator production in explants obtained at baseline.RA Patients were evaluated by disease activity score 28 joint C-reactive protein (DAS 28-, colour Doppler ultrasound (CDUS and 3 Tesla RA magnetic resonance imaging scores (RAMRIS. Explants were established from synovectomies from a needle arthroscopic procedure prior to initiation of bDMARD. Explants were incubated with the bDMARD in question, and the productions of interleukin-6 (IL-6, monocyte chemo-attractive protein-1 (MCP-1 and macrophage inflammatory protein-1-beta (MIP-1b were measured by multiplex immunoassays. The changes in clinical and imaging variables following a minimum of 3 months bDMARD treatment were compared to the baseline explant results. Mixed models and Spearman's rank correlations were performed. P-values below 0.05 were considered statistically significant.16 patients were included. IL-6 production in bDMARD-treated explants was significantly higher among clinical non-responders compared to responders (P = 0.04, and a lack of suppression of IL-6 by the bDMARDS correlated to a high DAS-28 (ρ = 0.57, P = 0.03, CDUS (ρ = 0.53, P = 0.04 and bone marrow oedema (ρ = 0.56, P = 0.03 at follow-up. No clinical association was found with explant MCP-1 production. MIP-1b could not be assessed due to a large number of samples below the detection limit.Synovial explants appear to deliver a disease-relevant output testing which when carried out in advance of bDMARD treatment can potentially pave the road for a more patient tailored treatment approach with better treatment effects.

  18. Optimally segmented magnetic structures

    DEFF Research Database (Denmark)

    Insinga, Andrea Roberto; Bahl, Christian; Bjørk, Rasmus

    We present a semi-analytical algorithm for magnet design problems, which calculates the optimal way to subdivide a given design region into uniformly magnetized segments.The availability of powerful rare-earth magnetic materials such as Nd-Fe-B has broadened the range of applications of permanent...... is not available.We will illustrate the results for magnet design problems from different areas, such as electric motors/generators (as the example in the picture), beam focusing for particle accelerators and magnetic refrigeration devices.......We present a semi-analytical algorithm for magnet design problems, which calculates the optimal way to subdivide a given design region into uniformly magnetized segments.The availability of powerful rare-earth magnetic materials such as Nd-Fe-B has broadened the range of applications of permanent...... magnets[1][2]. However, the powerful rare-earth magnets are generally expensive, so both the scientific and industrial communities have devoted a lot of effort into developing suitable design methods. Even so, many magnet optimization algorithms either are based on heuristic approaches[3...

  19. Plant regeneration from cotyledonary explants of Eucalyptus camaldulensis dehn and histological study of organogenesis in vitro

    Directory of Open Access Journals (Sweden)

    Roberson Dibax

    2010-04-01

    Full Text Available The present work aimed at regenerating plants of Eucalyptus camaldulensis from the cotyledonary explants and describing the anatomy of the tissues during callogenesis and organogenesis processes, in order to determine the origin of the buds. The cotyledonary leaves of E. camaldulensis were cultured in Murashige and Skoog (MS, WPM and JADS media supplemented with 2.7 µM NAA and 4.44 µM BAP. The best results for bud regeneration were obtained on MS and WPM media (57.5 and 55% of calluses formed buds, respectively. Shoot elongation and rooting (80% were obtained on MS/2 medium (with half-strength salt concentration with 0.2% activated charcoal. Acclimatization was performed in the growth chamber for 48 h and then the plants were transferred to a soil:vermiculite mixture and cultured in a greenhouse. Histological studies revealed that the callogenesis initiated in palisade parenchyma cells and that the adventitious buds were formed from the calluses, indicating indirect organogenesis.Este trabalho teve como objetivo a obtenção de plantas de Eucalyptus camaldulensis a partir de folhas cotiledonares e o estudo da anatomia dos tecidos durante a calogênese e organogênese para determinar a origem das gemas. Folhas cotiledonares foram cultivadas em meios de cultura MS, WPM e JADS suplementados com 2,7 µM de ANA e 4,44 µM de BAP. Os melhores resultados para a regeneração de gemas foram obtidos com os meios MS e WPM. Para o alongamento e enraizamento, o meio de cultura MS/2 contendo 0,2% de carvão ativado apresentou-se eficiente para ambas as etapas. A aclimatização foi realizada mediante a abertura dos frascos na sala de crescimento por 48 horas, seguido da transferência para casa-de-vegetação com nebulização intermitente. Estudos histológicos foram conduzidos e revelaram que a calogênese teve início nas células do parênquima paliçádico e que as gemas adventícias formaram-se a partir dos calos, indicando a organogênese indireta.

  20. In Vitro propagation of enterolobium cyclocarpum (guanacaste from nodal explants of axenic seedlings

    Directory of Open Access Journals (Sweden)

    Araceli Rodríguez Sahagún

    2007-01-01

    Full Text Available Enterolobium cyclocarpum (Jacq. Griseb. es un árbol leguminoso de uso múltiple, el cual es considerado una especie amenazada, resultado de la sobreexplotación y las bajas tasas de propagación natural debidas a las características intrínsecas del árbol. Una alternativa para superar este problema es el establecimiento de sistemas para su propagación masiva en tiempos cortos. En este trabajo, se investigó un protocolo para la propagación in vitro de E. cyclocarpum utilizando los segmentos nodales axénicos obtenidos de plántulas germinadas in vitro. Las semillas colectadas en dos comunidades mexicanas fueron germinadas tanto ex vitro como in vitro, y se evaluó el efecto de un pre-tratamiento de escarificación térmica. Para los experimentos de propagación se seleccionaron semillas provenientes de sólo una de las comunidades, debido a que presentaban una menor variabilidad genética de acuerdo con marcadores RAPD y a que existía una gran variación en las respuestas observadas en lotes de semillas mezclados. Esta variación fisiológica presente en semillas mezcladas, probablemente refleja un efecto del genotipo.Los segmentos nodales obtenidos de las plántulas fueron cultivados en medio basal MS suplementado con 30 g/L de sacarosa en presencia de distintas concentraciones de ácido 1-naftalenacético (ANA en combinación con benziladenina (BA o kinetina (KIN. La mayor tasa de multiplicación (de 4.75 brotes por explante en promedio se obtuvo cuando el medio MS fue suplementado con 2.2 µM BA y 10.7 µM ANA. Los brotes obtenidos fueron enraizados en medio MS con la mitad de concentración de sales y sin reguladores de crecimiento. Las plántulas micropropagadas fueron aclimatadas y transferidas exitosamente a suelo con una tasa de sobrevivencia del 90%. Estas plantas eran morfológicamente similares a la planta madre y no se detectó variación entre ellas por el uso de marcadores RAPD, lo cual hace posible el uso de este

  1. Inflammatory Response of Human Gestational Membranes to Ureaplasma parvum Using a Novel Dual-Chamber Tissue Explant System.

    Science.gov (United States)

    Potts, Lauren C; Feng, Liping; Seed, Patrick C; Jayes, Friederike L; Kuchibhatla, Maragatha; Antczak, Brian; Nazzal, Matthew K; Murtha, Amy P

    2016-05-01

    Preterm premature rupture of membranes (PPROM) is often associated with intra-amniotic inflammation and infection. Current understanding of the pathogenesis of PPROM includes activation of pro-inflammatory cytokines and proteolytic enzymes leading to compromise of membrane integrity. The impact of exposure to bacterial pathogens, including Ureaplasma parvum, on gestational membranes is poorly understood. Our objective was to develop a dual-chamber system to characterize the inflammatory response of gestational membranes to U. parvum in a directional nature. Full-thickness human gestational membrane explants, with either choriodecidua or amnion oriented superiorly, were suspended between two washers in a cylindrical device, creating two distinct compartments. Brilliant green dye was introduced into the top chamber to assess the integrity of the system. Tissue viability was evaluated after 72 h using a colorimetric cell proliferation assay. Choriodecidua or amnion was exposed to three doses of U. parvum and incubated for 24 h. Following treatment, media from each compartment were used for quantification of U. parvum (quantitative PCR), interleukin (IL)-8 (enzyme-linked immunosorbent assay), and matrix metalloproteinase (MMP)-2 and MMP-9 activity (zymography). We observed that system integrity and explant viability were maintained over 72 h. Dose-dependent increases in recovered U. parvum, IL-8 concentration, and MMP-2 activity were detected in both compartments. Significant differences in IL-8 concentration and MMP-9 activity were found between the choriodecidua and amnion. This tissue explant system can be used to investigate the inflammatory consequences of directional bacterial exposure for gestational membranes and provides insight into the pathogenesis of PPROM and infectious complications of pregnancy. © 2016 by the Society for the Study of Reproduction, Inc.

  2. Evaluation of novel biodegradable three-armed- and hyper-branched tissue adhesives in a meniscus explant model.

    Science.gov (United States)

    Bochyńska, A I; Hannink, G; Verhoeven, R; Grijpma, D W; Buma, P

    2017-05-01

    Current treatment methods to repair meniscal tears do not bring fully satisfactory results. Tissue adhesives are considered promising alternatives, since they are easy to apply and cause minimal tissue trauma. The first aim of this study was to analyze the adhesive properties of and tissue response to two recently developed biodegradable block copolymeric three-armed- and hyper-branched tissue adhesives. The second aim was to investigate if tissue surface modification with collagenase improves the attachment of the adhesives and increases the healing potential of the tissue. Cylindrical explants were harvested from bovine menisci. The central core of the explants was removed and glued back into the defect, with or without incubation in collagenase solution prior to gluing, using one of the novel glues, Dermabond® or fibrin glue. The repair constructs were cultured in vitro for 1 and 28 days. Adhesion tests and histology were performed to analyze the effects of the glue in combination with the additional treatment. The adhesive strength of the novel glues was 40-50 kPa, which was significantly higher than that of fibrin glue (15 kPa). Cells were present in direct contact with the glues, and the tissue remained vital during the whole culture period. Increased cellularity around the tear in the collagenase treated explants was observed after 1 day. The two newly developed tissue adhesives are attractive materials to be used for repair of meniscal tears. The beneficial influence of collagenase treatment in treating meniscal tears with glues still needs to be confirmed in more clinical relevant studies. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1405-1411, 2017. © 2017 Wiley Periodicals, Inc.

  3. Segmentation of the Infant Food Market

    OpenAIRE

    Hrůzová, Daniela

    2015-01-01

    The theoretical part covers general market segmentation, namely the marketing importance of differences among consumers, the essence of market segmentation, its main conditions and the process of segmentation, which consists of four consecutive phases - defining the market, determining important criteria, uncovering segments and developing segment profiles. The segmentation criteria, segmentation approaches, methods and techniques for the process of market segmentation are also described in t...

  4. Biosynthesis of intestinal microvillar proteins. Rapid expression of cytoskeletal components in microvilli of pig small intestinal mucosal explants

    DEFF Research Database (Denmark)

    Cowell, G M; Danielsen, E M

    1984-01-01

    Using alkaline extraction to separate cytoskeletal and membrane proteins of intestinal microvilli, the kinetics of assembly of these two microvillar protein compartments was studied by pulse-chase labelling of pig small intestinal mucosal explants, kept in organ culture. Following a 10 min pulse...... of [35S]methionine, the membrane proteins did not appear in the microvillar fraction until after 40-60 min of chase. In contrast, the cytoskeletal components, of which the 110-kDa protein and villin were immunologically identified, were expressed in the microvillar fraction immediately after the 10 min...

  5. Combination of MALDI-MSI and cassette dosing for evaluation of drug distribution in human skin explant

    DEFF Research Database (Denmark)

    Sørensen, Isabella S; Janfelt, Christian; Nielsen, Mette Marie B

    2017-01-01

    Study of skin penetration and distribution of the drug compounds in the skin is a major challenge in the development of topical drug products for treatment of skin diseases. It is crucial to have fast and efficacious screening methods which can provide information concerning the skin penetration ...... that combination of MALDI-MSI and cassette dosing can be used as a medium throughput screening tool at an early stage in the drug discovery/development process. Graphical abstract Investigation of drug distribution in human skin explant by MALDI-MSI after cassette dosing....

  6. Laparoscopic mesh explantation and drainage of sacral abscess remote from transvaginal excision of exposed sacral colpopexy mesh.

    Science.gov (United States)

    Roth, Ted M; Reight, Ian

    2012-07-01

    Sacral colpopexy may be complicated by mesh exposure, and the surgical treatment of mesh exposure typically results in minor postoperative morbidity and few delayed complications. A 75-year-old woman presented 7 years after a laparoscopic sacral colpopexy, with Mersilene mesh, with an apical mesh exposure. She underwent an uncomplicated transvaginal excision and was asymptomatic until 8 months later when she presented with vaginal drainage and a sacral abscess. This was successfully treated with laparoscopic enterolysis, drainage of the abscess, and explantation of the remaining mesh. Incomplete excision of exposed colpopexy mesh can lead to ascending infection and sacral abscess. Laparoscopic drainage and mesh removal may be considered in these patients.

  7. Phasing multi-segment undulators

    International Nuclear Information System (INIS)

    Chavanne, J.; Elleaume, P.; Vaerenbergh, P. Van

    1996-01-01

    An important issue in the manufacture of multi-segment undulators as a source of synchrotron radiation or as a free-electron laser (FEL) is the phasing between successive segments. The state of the art is briefly reviewed, after which a novel pure permanent magnet phasing section that is passive and does not require any current is presented. The phasing section allows the introduction of a 6 mm longitudinal gap between each segment, resulting in complete mechanical independence and reduced magnetic interaction between segments. The tolerance of the longitudinal positioning of one segment with respect to the next is found to be 2.8 times lower than that of conventional phasing. The spectrum at all gaps and useful harmonics is almost unchanged when compared with a single-segment undulator of the same total length. (au) 3 refs

  8. The LOFT Ground Segment

    DEFF Research Database (Denmark)

    Bozzo, E.; Antonelli, A.; Argan, A.

    2014-01-01

    targets per orbit (~90 minutes), providing roughly ~80 GB of proprietary data per day (the proprietary period will be 12 months). The WFM continuously monitors about 1/3 of the sky at a time and provides data for about ~100 sources a day, resulting in a total of ~20 GB of additional telemetry. The LOFT...... Burst alert System additionally identifies on-board bright impulsive events (e.g., Gamma-ray Bursts, GRBs) and broadcasts the corresponding position and trigger time to the ground using a dedicated system of ~15 VHF receivers. All WFM data are planned to be made public immediately. In this contribution...... we summarize the planned organization of the LOFT ground segment (GS), as established in the mission Yellow Book 1 . We describe the expected GS contributions from ESA and the LOFT consortium. A review is provided of the planned LOFT data products and the details of the data flow, archiving...

  9. Segmented heat exchanger

    Science.gov (United States)

    Baldwin, Darryl Dean; Willi, Martin Leo; Fiveland, Scott Byron; Timmons, Kristine Ann

    2010-12-14

    A segmented heat exchanger system for transferring heat energy from an exhaust fluid to a working fluid. The heat exchanger system may include a first heat exchanger for receiving incoming working fluid and the exhaust fluid. The working fluid and exhaust fluid may travel through at least a portion of the first heat exchanger in a parallel flow configuration. In addition, the heat exchanger system may include a second heat exchanger for receiving working fluid from the first heat exchanger and exhaust fluid from a third heat exchanger. The working fluid and exhaust fluid may travel through at least a portion of the second heat exchanger in a counter flow configuration. Furthermore, the heat exchanger system may include a third heat exchanger for receiving working fluid from the second heat exchanger and exhaust fluid from the first heat exchanger. The working fluid and exhaust fluid may travel through at least a portion of the third heat exchanger in a parallel flow configuration.

  10. International EUREKA: Market Segment

    International Nuclear Information System (INIS)

    1982-03-01

    The purpose of the Market Segment of the EUREKA model is to simultaneously project uranium market prices, uranium supply and purchasing activities. The regional demands are extrinsic. However, annual forward contracting activities to meet these demands as well as inventory requirements are calculated. The annual price forecast is based on relatively short term, forward balances between available supply and desired purchases. The forecasted prices and extrapolated price trends determine decisions related to exploration and development, new production operations, and the operation of existing capacity. Purchasing and inventory requirements are also adjusted based on anticipated prices. The calculation proceeds one year at a time. Conditions calculated at the end of one year become the starting conditions for the calculation in the subsequent year

  11. Probabilistic retinal vessel segmentation

    Science.gov (United States)

    Wu, Chang-Hua; Agam, Gady

    2007-03-01

    Optic fundus assessment is widely used for diagnosing vascular and non-vascular pathology. Inspection of the retinal vasculature may reveal hypertension, diabetes, arteriosclerosis, cardiovascular disease and stroke. Due to various imaging conditions retinal images may be degraded. Consequently, the enhancement of such images and vessels in them is an important task with direct clinical applications. We propose a novel technique for vessel enhancement in retinal images that is capable of enhancing vessel junctions in addition to linear vessel segments. This is an extension of vessel filters we have previously developed for vessel enhancement in thoracic CT scans. The proposed approach is based on probabilistic models which can discern vessels and junctions. Evaluation shows the proposed filter is better than several known techniques and is comparable to the state of the art when evaluated on a standard dataset. A ridge-based vessel tracking process is applied on the enhanced image to demonstrate the effectiveness of the enhancement filter.

  12. Effects of gamma-rays irradiation in seed of mungbean (vigna radiata (L.) wilczek) composition of media on shoot regeneration of explants from node of cotyledon

    International Nuclear Information System (INIS)

    Hutabarat, Dameria; H, Soeranto

    1998-01-01

    Study the effects of gamma-rays irradiation and composition and media on shoot regeneration of explants from node of cotyledon of mungbean. Wallet variety have been conducted. The explants derived of irradiated seeds of 10-20 Gy of gamma rays were planted in the 0.7% agar solution. One day after planting in the agar media the embryo axis of germinate seed were removed and the node of cotyledon were cultured in the regeneration media as examples. The results shown that shoot regeneration was influenced by media composition and the doses of gamma rays irradiation in seed. In the MURASHIGE and SKOOG medium which contain of BAP or 2-iP or Kinetin with 3 ppm concentrate respectively the explants could produced 100% of shoots. However, the highest. number of produced shoot (3 shoots) was showed in the medium which contained of BAP. The medium with I ppm concentrate od BAP could produced 100% shoot regeneration and the maximum number of shoots (4 shoots) per explant was showed in with 5 ppm. concentrate of BAP. The effectivity off BAP for shoot regeneration by enrichment of 12 ppm Ag 2 SO 4 in the media. Irradiation of 10-20 Gy gamma rays on seeds of mungbean walet variety could improved shoot regeneration of explants from node cotyledon. (author)

  13. Segmented rail linear induction motor

    Science.gov (United States)

    Cowan, Jr., Maynard; Marder, Barry M.

    1996-01-01

    A segmented rail linear induction motor has a segmented rail consisting of a plurality of nonferrous electrically conductive segments aligned along a guideway. The motor further includes a carriage including at least one pair of opposed coils fastened to the carriage for moving the carriage. A power source applies an electric current to the coils to induce currents in the conductive surfaces to repel the coils from adjacent edges of the conductive surfaces.

  14. Segmentation-DrivenTomographic Reconstruction

    DEFF Research Database (Denmark)

    Kongskov, Rasmus Dalgas

    such that the segmentation subsequently can be carried out by use of a simple segmentation method, for instance just a thresholding method. We tested the advantages of going from a two-stage reconstruction method to a one stage segmentation-driven reconstruction method for the phase contrast tomography reconstruction......The tomographic reconstruction problem is concerned with creating a model of the interior of an object from some measured data, typically projections of the object. After reconstructing an object it is often desired to segment it, either automatically or manually. For computed tomography (CT...

  15. Automated medical image segmentation techniques

    Directory of Open Access Journals (Sweden)

    Sharma Neeraj

    2010-01-01

    Full Text Available Accurate segmentation of medical images is a key step in contouring during radiotherapy planning. Computed topography (CT and Magnetic resonance (MR imaging are the most widely used radiographic techniques in diagnosis, clinical studies and treatment planning. This review provides details of automated segmentation methods, specifically discussed in the context of CT and MR images. The motive is to discuss the problems encountered in segmentation of CT and MR images, and the relative merits and limitations of methods currently available for segmentation of medical images.

  16. ADVANCED CLUSTER BASED IMAGE SEGMENTATION

    Directory of Open Access Journals (Sweden)

    D. Kesavaraja

    2011-11-01

    Full Text Available This paper presents efficient and portable implementations of a useful image segmentation technique which makes use of the faster and a variant of the conventional connected components algorithm which we call parallel Components. In the Modern world majority of the doctors are need image segmentation as the service for various purposes and also they expect this system is run faster and secure. Usually Image segmentation Algorithms are not working faster. In spite of several ongoing researches in Conventional Segmentation and its Algorithms might not be able to run faster. So we propose a cluster computing environment for parallel image Segmentation to provide faster result. This paper is the real time implementation of Distributed Image Segmentation in Clustering of Nodes. We demonstrate the effectiveness and feasibility of our method on a set of Medical CT Scan Images. Our general framework is a single address space, distributed memory programming model. We use efficient techniques for distributing and coalescing data as well as efficient combinations of task and data parallelism. The image segmentation algorithm makes use of an efficient cluster process which uses a novel approach for parallel merging. Our experimental results are consistent with the theoretical analysis and practical results. It provides the faster execution time for segmentation, when compared with Conventional method. Our test data is different CT scan images from the Medical database. More efficient implementations of Image Segmentation will likely result in even faster execution times.

  17. Effects of low molecular weight hyaluronan combined with carprofen on canine osteoarthritis articular chondrocytes and cartilage explants in vitro.

    Science.gov (United States)

    Euppayo, Thippaporn; Siengdee, Puntita; Buddhachat, Kittisak; Pradit, Waranee; Viriyakhasem, Nawarat; Chomdej, Siriwadee; Ongchai, Siriwan; Harada, Yasuji; Nganvongpanit, Korakot

    2015-09-01

    Intra-articular injection with non-steroidal anti-inflammatory drugs (NSAIDs) is used to treat inflammatory joint disease, but the side effects of NSAIDs include chondrotoxicity. Hyaluronan has shown positive effects on chondrocytes by reducing apoptosis and increasing proteoglycan synthesis. The purposes of this study were to evaluate the effects of low molecular weight hyaluronan (low MW HA), carprofen 25 mg/ml, carprofen 12.5 mg/ml, and a combination of HA and carprofen on canine osteoarthritis (OA) articular chondrocytes and a cartilage explant model in terms of cell viability, extracellular matrix remaining, and gene expression after exposure. In chondrocyte culture, MTT assay was used to evaluate the chondrotoxicity of IC50 and IC80 of carprofen with HA. In cartilage explant culture, two kinds of extracellular matrix (uronic acid and collagen) remaining in cartilage were used to evaluate cartilage damage for 14 d after treatment. Expression of COL2A1, AGG, and MMP3 was used to evaluate the synthesis and degradation of the matrix for 7 d after treatment. In chondrocyte culture, low MW HA could preserve OA chondrocyte viability but could not reduce the chondrotoxicity level of carprofen (P carprofen caused less destruction of uronic acid and collagen structure when compared with the control (P carprofen resulted in higher COL2A1 and AGG expression levels than carprofen alone.

  18. Plant Explants Grown on Medium Supplemented with Fe3O4 Nanoparticles Have a Significant Increase in Embryogenesis

    Directory of Open Access Journals (Sweden)

    Inese Kokina

    2017-01-01

    Full Text Available Development of nanotechnology leads to the increasing release of nanoparticles in the environment that results in accumulation of different NPs in living organisms including plants. This can lead to serious changes in plant cultures which leads to genotoxicity. The aims of the present study were to detect if iron oxide NPs pass through the flax cell wall, to compare callus morphology, and to estimate the genotoxicity in Linum usitatissimum L. callus cultures induced by different concentrations of Fe3O4 nanoparticles. Two parallel experiments were performed: experiment A, where flax explants were grown on medium supplemented with 0.5 mg/l, 1 mg/l, and 1.5 mg/l Fe3O4 NPs for callus culture obtaining, and experiment B, where calluses obtained from basal MS medium were transported into medium supplemented with concentrations of NPs identical to experiment A. Obtained results demonstrate similarly in both experiments that 25 nm Fe3O4 NPs pass into callus cells and induce low toxicity level in the callus cultures. Nevertheless, calluses from experiment A showed 100% embryogenesis in comparison with experiment B where 100% rhizogenesis was noticed. It could be associated with different stress levels and adaptation time for explants and calluses that were transported into medium with Fe3O4 NPs supplementation.

  19. 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)

  20. Cellular, pharmacological, and biophysical evaluation of explanted lungs from a patient with sickle cell disease and severe pulmonary arterial hypertension.

    Science.gov (United States)

    Rogers, Natasha M; Yao, Mingyi; Sembrat, John; George, M Patricia; Knupp, Heather; Ross, Mark; Sharifi-Sanjani, Maryam; Milosevic, Jadranka; St Croix, Claudette; Rajkumar, Revathi; Frid, Maria G; Hunter, Kendall S; Mazzaro, Luciano; Novelli, Enrico M; Stenmark, Kurt R; Gladwin, Mark T; Ahmad, Ferhaan; Champion, Hunter C; Isenberg, Jeffrey S

    2013-12-01

    Pulmonary hypertension is recognized as a leading cause of morbidity and mortality in patients with sickle cell disease (SCD). We now report benchtop phenotyping from the explanted lungs of the first successful lung transplant in SCD. Pulmonary artery smooth muscle cells (PASMCs) cultured from the explanted lungs were analyzed for proliferate capacity, superoxide (O2 (•-)) production, and changes in key pulmonary arterial hypertension (PAH)-associated molecules and compared with non-PAH PASMCs. Upregulation of several pathologic processes persisted in culture in SCD lung PASMCs in spite of cell passage. SCD lung PASMCs showed growth factor- and serum-independent proliferation, upregulation of matrix genes, and increased O2 (•-) production compared with control cells. Histologic analysis of SCD-associated PAH arteries demonstrated increased and ectopically located extracellular matrix deposition and degradation of elastin fibers. Biomechanical analysis of these vessels confirmed increased arterial stiffening and loss of elasticity. Functional analysis of distal fifth-order pulmonary arteries from these lungs demonstrated increased vasoconstriction to an α1-adrenergic receptor agonist and concurrent loss of both endothelial-dependent and endothelial-independent vasodilation compared with normal pulmonary arteries. This is the first study to evaluate the molecular, cellular, functional, and mechanical changes in end-stage SCD-associated PAH.

  1. Inhibition of HIV transmission in human cervicovaginal explants and humanized mice using CD4 aptamer-siRNA chimeras

    Science.gov (United States)

    Wheeler, Lee Adam; Trifonova, Radiana; Vrbanac, Vladimir; Basar, Emre; McKernan, Shannon; Xu, Zhan; Seung, Edward; Deruaz, Maud; Dudek, Tim; Einarsson, Jon Ivar; Yang, Linda; Allen, Todd M.; Luster, Andrew D.; Tager, Andrew M.; Dykxhoorn, Derek M.; Lieberman, Judy

    2011-01-01

    The continued spread of the HIV epidemic underscores the need to interrupt transmission. One attractive strategy is a topical vaginal microbicide. Sexual transmission of herpes simplex virus type 2 (HSV-2) in mice can be inhibited by intravaginal siRNA application. To overcome the challenges of knocking down gene expression in immune cells susceptible to HIV infection, we used chimeric RNAs composed of an aptamer fused to an siRNA for targeted gene knockdown in cells bearing an aptamer-binding receptor. Here, we showed that CD4 aptamer-siRNA chimeras (CD4-AsiCs) specifically suppress gene expression in CD4+ T cells and macrophages in vitro, in polarized cervicovaginal tissue explants, and in the female genital tract of humanized mice. CD4-AsiCs do not activate lymphocytes or stimulate innate immunity. CD4-AsiCs that knock down HIV genes and/or CCR5 inhibited HIV infection in vitro and in tissue explants. When applied intravaginally to humanized mice, CD4-AsiCs protected against HIV vaginal transmission. Thus, CD4-AsiCs could be used as the active ingredient of a microbicide to prevent HIV sexual transmission. PMID:21576818

  2. MRI screening for silicone breast implant rupture: accuracy, inter- and intraobserver variability using explantation results as reference standard

    Energy Technology Data Exchange (ETDEWEB)

    Maijers, M.C.; Ritt, M.J.P.F. [VU University Medical Centre, Department of Plastic, Reconstructive and Hand Surgery, De Boelelaan 1117, PO Box 7057, Amsterdam (Netherlands); Niessen, F.B. [VU University Medical Centre, Department of Plastic, Reconstructive and Hand Surgery, De Boelelaan 1117, PO Box 7057, Amsterdam (Netherlands); Jan van Goyen Clinic, Department of Plastic Surgery, Amsterdam (Netherlands); Veldhuizen, J.F.H. [MRI Centre, Amsterdam (Netherlands); Manoliu, R.A. [MRI Centre, Amsterdam (Netherlands); VU University Medical Centre, Department of Radiology, Amsterdam (Netherlands)

    2014-06-15

    The recall of Poly Implant Prothese (PIP) silicone breast implants in 2010 resulted in large numbers of asymptomatic women with implants who underwent magnetic resonance imaging (MRI) screening. This study's aim was to assess the accuracy and interobserver variability of MRI screening in the detection of rupture and extracapsular silicone leakage. A prospective study included 107 women with 214 PIP implants who underwent explantation preceded by MRI. In 2013, two radiologists blinded for previous MRI findings or outcome at surgery, independently re-evaluated all MRI examinations. A structured protocol described the MRI findings. The ex vivo findings served as reference standard. In 208 of the 214 explanted prostheses, radiologists agreed independently about the condition of the implants. In five of the six cases they disagreed (2.6 %), but subsequently reached consensus. A sensitivity of 93 %, specificity of 93 %, positive predictive value of 77 % and negative predictive value of 98 % was found. The interobserver agreement was excellent (kappa value of 0.92). MRI has a high accuracy in diagnosing rupture in silicone breast implants. Considering the high kappa value of interobserver agreement, MRI appears to be a consistent diagnostic test. A simple, uniform classification, may improve communication between radiologist and plastic surgeon. (orig.)

  3. MRI screening for silicone breast implant rupture: accuracy, inter- and intraobserver variability using explantation results as reference standard

    International Nuclear Information System (INIS)

    Maijers, M.C.; Ritt, M.J.P.F.; Niessen, F.B.; Veldhuizen, J.F.H.; Manoliu, R.A.

    2014-01-01

    The recall of Poly Implant Prothese (PIP) silicone breast implants in 2010 resulted in large numbers of asymptomatic women with implants who underwent magnetic resonance imaging (MRI) screening. This study's aim was to assess the accuracy and interobserver variability of MRI screening in the detection of rupture and extracapsular silicone leakage. A prospective study included 107 women with 214 PIP implants who underwent explantation preceded by MRI. In 2013, two radiologists blinded for previous MRI findings or outcome at surgery, independently re-evaluated all MRI examinations. A structured protocol described the MRI findings. The ex vivo findings served as reference standard. In 208 of the 214 explanted prostheses, radiologists agreed independently about the condition of the implants. In five of the six cases they disagreed (2.6 %), but subsequently reached consensus. A sensitivity of 93 %, specificity of 93 %, positive predictive value of 77 % and negative predictive value of 98 % was found. The interobserver agreement was excellent (kappa value of 0.92). MRI has a high accuracy in diagnosing rupture in silicone breast implants. Considering the high kappa value of interobserver agreement, MRI appears to be a consistent diagnostic test. A simple, uniform classification, may improve communication between radiologist and plastic surgeon. (orig.)

  4. İn Vitro Koşullarda Domates (Lycopersicon esculentum Mill. Bitkisinde Hipokotil ve Kotiledon Eksplantlarından Kallus ve Sürgün Oluşumu

    Directory of Open Access Journals (Sweden)

    Emel YILMAZ

    2015-01-01

    Full Text Available In this study, the effects of various plant growth regulators and culture conditions on the callus induction and shoot regeneration in M-28 hybrid cultivar of tomato have been evaluated. The seeds were germinated in half strength (½ Murashige-Skoog (MS medium and the cotyledon (5x5 mm and hypocotyl (5 mm segments of the 10 day old seedlings were cultured on semi-solide MS medium containing different concentrations and combinations of plant growth regulators. The best result on callus formation, both explants (cotyledon and hypocotyl and both culture conditions (photoperiodic condition at 16 hours light8 hours dark ve 25 days dark condition after photoperiodic condition, was found MS medium supplemented with 1 mg/l BAP+1 mg/l NAA (%100 callus formation. At the adventitious shoot induction, under dark condition after photoperiodic condition, the best results were obtained from MS medium supplemented with 2 mg/l Kin+0.2 mg/l IAA, only 2 mg/l BAP and only 3 mg/l TDZ and both explants (regenerated explant from %72 to %100. The highest number of shoots regenerated per explant (17.30±5.72 number, was found under dark condition after photoperiodic condition at MS medium supplemented with TDZ of cotyledon explants. The highest adventitious shoot length (33.00±0.57 mm and the number of leaf in shoot (3.00±0.00 number were obtained at MS supplemented with 1 mg/l BAP+1 mg/l NAA of hypocotyl explants and photoperiodic conditions

  5. Histological analysis of the callogenesis and organogenesis from root segments of Curcuma zedoaria Roscoe

    Directory of Open Access Journals (Sweden)

    Marcia O. Mello

    2001-06-01

    Full Text Available Callus was induced from root segments taken from in vitro grown plants of Curcuma zedoaria Roscoe. The explants were cultured on agar-solidified Murashige and Skoog medium supplemented with 13.4muM of alpha-naphthaleneacetic acid and 2.2muM of 6-benzylaminopurine at 25ºC in the dark. Histological analysis revealed that callus was formed from the hypertrophied cortical parenchyma cells of the explant. Some of these cells underwent division while the surrounding cells accumulated starch. Callus was capable of shoot bud regeneration after 70 days when it was transfered to liquid medium of the same composition. After 30 days in liquid medium, buds developed from nodular structures. The adventitious shoots developed extensive root systems when they were placed on agar-solidified Murashige and Skoog medium without growth regulators at 25º C in the light. The establishment of these plantlets in soil was about 95%.Calo de Curcuma zedoaria Roscoe foram induzidos a partir de segmentos de raízes de plantas cultivadas in vitro. Os explantes foram inoculados em meio de Murashige & Skoog solidificado com ágar e suplementado com 13,4miM de ácido alfa-naftaleno acético e 2,2miM de 6-benzilaminopurina e mantidos no escuro a 25°C. As análises histológicas realizadas revelaram que os callus eram formados a partir de células hipertrofiadas do parênquima cortical do explante. Algumas destas células entravam em divisão, enquanto as células vizinhas a estas acumulavam amido. Após 70 dias, calos transferidos para meio de Murashige & Skoog líquido de mesma composição, eram capazes de regenerar plantas. Após 30 dias em meio líquido, gemas se desenvolveram de estruturas nodulares. Estas gemas adventíceas formaram um abundante sistema radicular quando transferidas para meio de Murashige & Skoog solidificado com ágar, sem regulador de crescimento e mantidas a 25°C na luz. A taxa de sobrevivência das plantas foi de 95%.

  6. Isolation of Highly Purified Fractions of Plasma Membrane and Tonoplast from the Same Homogenate of Soybean Hypocotyls by Free-Flow Electrophoresis 1

    Science.gov (United States)

    Sandelius, Anna Stina; Penel, Claude; Auderset, Guy; Brightman, Andrew; Millard, Merle; Morré, D. James

    1986-01-01

    A procedure is described whereby highly purified fractions of plasma membrane and tonoplast were isolated from hypocotyls of dark-grown soybean (Glycine max L. var Wayne) by the technique of preparative free-flow electrophoresis. Fractions migrating the slowest toward the anode were enriched in thick (10 nanometers) membranes identified as plasma membranes based on ability to bind N-1-naphthylphthalamic acid (NPA), glucan synthetase-II, and K+-stimulated, vanadate-inhibited Mg2+ ATPase, reaction with phosphotungstic acid at low pH on electron microscope sections, and morphological evaluations. Fractions migrating farthest toward the anode (farthest from the point of sample injection) were enriched in membrane vesicles with thick (7-9 nanometers) membranes that did not stain with phosphotungstic acid at low pH, contained a nitrate-inhibited, Cl-stimulated ATPase and had the in situ morphological characteristics of tonoplast including the presence of flocculent contents. These vesicles neither bound NPA nor contained levels of glucan synthetase II above background. Other membranous cell components such as dictyosomes (fucosyltransferase, latent nucleosidediphosphate phosphatase), endoplasmic reticulum vesicles (NADH- and NADPH- cytochrome c reductase), mitochondria (succinate-2(p-indophenyl)-3-p-nitrophenyl)-5-phenyl tetrazolium-reductase and cytochrome oxidase) and plastids (carotenoids and monogalactosyl diglyceride synthetase) were identified on the basis of appropriate marker constituents and, except for plastid thylakoids, had thin (marker activities. From electron microscope morphometry (using both membrane measurements and staining with phosphotungstic acid at low pH) and analysis of marker enzymes, both plasma membrane and tonoplast fractions were estimated to be about 90% pure. Neither fraction appeared to be contaminated by the other by more than 3%. Images Fig. 1 Fig. 3 Fig. 4 Fig. 5 Fig. 9 PMID:16664771

  7. Microculture of western white pine (Pinus monticola) by induction of shoots on bud explants from 1- to 7-year-old trees.

    Science.gov (United States)

    Lapp, M S; Malinek, J; Coffey, M

    1996-04-01

    We developed a protocol for the production of shoots from bud explants from 1- to 7-year-old trees of western white pine (Pinus monticola Dougl.). The best explant was a 2-mm-thick cross-sectional slice of the early winter bud. Genotype of the donor tree was a significant factor affecting shoot production, but more than 80% of the genotypes tested produced shoots. Of the media tested, bud slices from 1- to 3-year-old trees grew best in Litvay's medium containing N(6)-benzyladenine in the range of 1 to 30 micro M, whereas bud slices from older trees grew best in Gupta and Durzan's DCR medium with zeatin riboside. Up to 400 shoots more than 3 mm in height were obtained from 100 bud-slice explants taken from 7-year-old western white pine trees.

  8. [Influence of genotype, explant type and component of culture medium on in vitro callus induction and shoot organogenesis of tomato (Solanum lycopersicum L.)].

    Science.gov (United States)

    Khaliluev, M R; Bogoutdinova, L R; Baranova, G B; Baranova, E N; Kharchenko, P N; Dolgov, S V

    2014-01-01

    The influence of explant type as well as of the type of growth regulators and concentration on callus induction processes and somatic organogenesis of shoots was studied in vitro on four tomato genotypes of Russian breeding. Cytological study of callus tissue was conducted. It was established that tomato varieties possess a substantially greater ability to indirect shoot organogenesis compared with the F1 hybrid. The highest frequency of somatic organogenesis of shoots, as well as their number per explant, was observed for most of the genotypes studied during the cultivation of cotyledons on Murashige-Skoog culture medium containing 2 mg/l of zeatin in combination with 0.1 mg/l of 3-indoleacetic acid. An effective protocol of indirect somatic organogenesis of shoots from different explants of tomato varieties with a frequency of more than 80% was developed.

  9. Region segmentation along image sequence

    International Nuclear Information System (INIS)

    Monchal, L.; Aubry, P.

    1995-01-01

    A method to extract regions in sequence of images is proposed. Regions are not matched from one image to the following one. The result of a region segmentation is used as an initialization to segment the following and image to track the region along the sequence. The image sequence is exploited as a spatio-temporal event. (authors). 12 refs., 8 figs

  10. Market segmentation using perceived constraints

    Science.gov (United States)

    Jinhee Jun; Gerard Kyle; Andrew Mowen

    2008-01-01

    We examined the practical utility of segmenting potential visitors to Cleveland Metroparks using their constraint profiles. Our analysis identified three segments based on their scores on the dimensions of constraints: Other priorities--visitors who scored the highest on 'other priorities' dimension; Highly Constrained--visitors who scored relatively high on...

  11. Market Segmentation: An Instructional Module.

    Science.gov (United States)

    Wright, Peter H.

    A concept-based introduction to market segmentation is provided in this instructional module for undergraduate and graduate transportation-related courses. The material can be used in many disciplines including engineering, business, marketing, and technology. The concept of market segmentation is primarily a transportation planning technique by…

  12. IFRS 8 – OPERATING SEGMENTS

    Directory of Open Access Journals (Sweden)

    BOCHIS LEONICA

    2009-05-01

    Full Text Available Segment reporting in accordance with IFRS 8 will be mandatory for annual financial statements covering periods beginning on or after 1 January 2009. The standards replaces IAS 14, Segment Reporting, from that date. The objective of IFRS 8 is to require

  13. Reduplication Facilitates Early Word Segmentation

    Science.gov (United States)

    Ota, Mitsuhiko; Skarabela, Barbora

    2018-01-01

    This study explores the possibility that early word segmentation is aided by infants' tendency to segment words with repeated syllables ("reduplication"). Twenty-four nine-month-olds were familiarized with passages containing one novel reduplicated word and one novel non-reduplicated word. Their central fixation times in response to…

  14. The Importance of Marketing Segmentation

    Science.gov (United States)

    Martin, Gillian

    2011-01-01

    The rationale behind marketing segmentation is to allow businesses to focus on their consumers' behaviors and purchasing patterns. If done effectively, marketing segmentation allows an organization to achieve its highest return on investment (ROI) in turn for its marketing and sales expenses. If an organization markets its products or services to…

  15. Essays in international market segmentation

    NARCIS (Netherlands)

    Hofstede, ter F.

    1999-01-01

    The primary objective of this thesis is to develop and validate new methodologies to improve the effectiveness of international segmentation strategies. The current status of international market segmentation research is reviewed in an introductory chapter, which provided a number of

  16. Agrobacterium-mediated genetic transformation of Pogostemon cablin (Blanco) Benth. Using leaf explants: bactericidal effect of leaf extracts and counteracting strategies.

    Science.gov (United States)

    Paul, Anamika; Bakshi, Souvika; Sahoo, Debee Prasad; Kalita, Mohan Chandra; Sahoo, Lingaraj

    2012-04-01

    An optimized protocol for Agrobacterium tumefaciens-mediated transformation of patchouli using leaf disk explants is reported. In vitro antibacterial activity of leaf extracts of the plants revealed Agrobacterium sensitivity to the extracts. Fluorometric assay of bacterial cell viability indicated dose-dependent cytotoxic activity of callus extract against Agrobacterium cells. Addition of 0.1% Tween 20 and 2 g/l L-glutamine to Agrobacterium infection medium counteracted the bactericidal effect and significantly increased the T-DNA delivery to explants. A short preculture of explants for 2 days followed by infection with Agrobacterium in medium containing 150 μM of acetosyringone were found essential for efficient T-DNA delivery. Cocultivation for 3 days at 22 °C in conjunction with other optimized factors resulted in maximum T-DNA delivery. The Agrobacterium-mediated transformation of leaf disk explants were found significantly related to physiological age of the explants, age and origin of the of the donor plant. Leaf explants from second node of the 3-month-old in vivo plants showed highest transformation efficiency (94.3%) revealed by transient GUS expression assay. Plants selected on medium containing 20 mg/l kanamycin showed stable GUS expression in leaves and stem. The elongated shoots readily developed roots on kanamycin-free rooting medium and on transfer to soil, plants were successfully established. Polymerase chain reaction (PCR) and reverse-transcriptase PCR analysis in putative plants confirmed their transgenic nature. The established transformation method should provide new opportunities for the genetic improvement of patchouli for desirable trait.

  17. Complications and Short-Term Explantation Rate Following Artificial Urinary Sphincter Implantation: Results from a Large Middle European Multi-Institutional Case Series.

    Science.gov (United States)

    Kretschmer, Alexander; Hüsch, Tanja; Thomsen, Frauke; Kronlachner, Dominik; Obaje, Alice; Anding, Ralf; Pottek, Tobias; Rose, Achim; Olianas, Roberto; Friedl, Alexander; Hübner, Wilhelm; Homberg, Roland; Pfitzenmaier, Jesco; Grein, Ulrich; Queissert, Fabian; Naumann, Carsten Maik; Schweiger, Josef; Wotzka, Carola; Nyarangi-Dix, Joanne N; Hofmann, Torben; Seiler, Roland; Haferkamp, Axel; Bauer, Ricarda M

    2016-01-01

    Background/Aims/Objectives: To analyze perioperative complication and short-term explantation rates after perineal or penoscrotal single-cuff and double-cuff artificial urinary sphincter (AUS) implantation in a large middle European multi-institutional patient cohort. 467 male patients with stress urinary incontinence underwent implantation of a perineal single-cuff (n = 152), penoscrotal single-cuff (n = 99), or perineal double-cuff (n = 216) AUS between 2010 and 2012. Postoperative complications and 6-month explantation rates were assessed. For statistical analysis, Fisher's exact test and Kruskal-Wallis rank sum test, and a multiple logistic regression model were used (p AUS, penoscrotal single-cuff implantation led to significantly increased short-term explantation rates (8.6% (perineal) vs. 19.2% (penoscrotal), p = 0.019). The postoperative infection rate was significantly higher after double-cuff compared to single-cuff implantation (6.0% (single-cuff) vs. 13.9% (double-cuff), p = 0.019). The short-term explantation rate after primary double-cuff placement was 6.5% (p = 0.543 vs. perineal single-cuff). In multivariate analysis, the penoscrotal approach (p = 0.004), intraoperative complications (p = 0.005), postoperative bleeding (p = 0.011), and perioperative infection (p < 0.001) were independent risk factors for short-term explantation. Providing data from a large contemporary multi-institutional patient cohort from high-volume and low-volume institutions, our results reflect the current standard of care in middle Europe. We indicate that the penoscrotal approach is an independent risk factor for increased short-term explantation rates. © 2016 S. Karger AG, Basel.

  18. Segmental vitiligo with segmental morphea: An autoimmune link?

    Directory of Open Access Journals (Sweden)

    Pravesh Yadav

    2014-01-01

    Full Text Available An 18-year old girl with segmental vitiligo involving the left side of the trunk and left upper limb with segmental morphea involving the right side of trunk and right upper limb without any deeper involvement is illustrated. There was no history of preceding drug intake, vaccination, trauma, radiation therapy, infection, or hormonal therapy. Family history of stable vitiligo in her brother and a history of type II diabetes mellitus in the father were elicited. Screening for autoimmune diseases and antithyroid antibody was negative. An autoimmune link explaining the co-occurrence has been proposed. Cutaneous mosiacism could explain the presence of both the pathologies in a segmental distribution.

  19. Using Predictability for Lexical Segmentation.

    Science.gov (United States)

    Çöltekin, Çağrı

    2017-09-01

    This study investigates a strategy based on predictability of consecutive sub-lexical units in learning to segment a continuous speech stream into lexical units using computational modeling and simulations. Lexical segmentation is one of the early challenges during language acquisition, and it has been studied extensively through psycholinguistic experiments as well as computational methods. However, despite strong empirical evidence, the explicit use of predictability of basic sub-lexical units in models of segmentation is underexplored. This paper presents an incremental computational model of lexical segmentation for exploring the usefulness of predictability for lexical segmentation. We show that the predictability cue is a strong cue for segmentation. Contrary to earlier reports in the literature, the strategy yields state-of-the-art segmentation performance with an incremental computational model that uses only this particular cue in a cognitively plausible setting. The paper also reports an in-depth analysis of the model, investigating the conditions affecting the usefulness of the strategy. Copyright © 2016 Cognitive Science Society, Inc.

  20. The Hierarchy of Segment Reports

    Directory of Open Access Journals (Sweden)

    Danilo Dorović

    2015-05-01

    Full Text Available The article presents an attempt to find the connection between reports created for managers responsible for different business segments. With this purpose, the hierarchy of the business reporting segments is proposed. This can lead to better understanding of the expenses under common responsibility of more than one manager since these expenses should be in more than one report. The structure of cost defined per business segment hierarchy with the aim of new, unusual but relevant cost structure for management can be established. Both could potentially bring new information benefits for management in the context of profit reporting.

  1. Segmental dilatation of the ileum

    Directory of Open Access Journals (Sweden)

    Tune-Yie Shih

    2017-01-01

    Full Text Available A 2-year-old boy was sent to the emergency department with the chief problem of abdominal pain for 1 day. He was just discharged from the pediatric ward with the diagnosis of mycoplasmal pneumonia and paralytic ileus. After initial examinations and radiographic investigations, midgut volvulus was impressed. An emergency laparotomy was performed. Segmental dilatation of the ileum with volvulus was found. The operative procedure was resection of the dilated ileal segment with anastomosis. The postoperative recovery was uneventful. The unique abnormality of gastrointestinal tract – segmental dilatation of the ileum, is described in details and the literature is reviewed.

  2. Accounting for segment correlations in segmented gamma-ray scans

    International Nuclear Information System (INIS)

    Sheppard, G.A.; Prettyman, T.H.; Piquette, E.C.

    1994-01-01

    In a typical segmented gamma-ray scanner (SGS), the detector's field of view is collimated so that a complete horizontal slice or segment of the desired thickness is visible. Ordinarily, the collimator is not deep enough to exclude gamma rays emitted from sample volumes above and below the segment aligned with the collimator. This can lead to assay biases, particularly for certain radioactive-material distributions. Another consequence of the collimator's low aspect ratio is that segment assays at the top and bottom of the sample are biased low because the detector's field of view is not filled. This effect is ordinarily countered by placing the sample on a low-Z pedestal and scanning one or more segment thicknesses below and above the sample. This takes extra time, however, We have investigated a number of techniques that both account for correlated segments and correct for end effects in SGS assays. Also, we have developed an algorithm that facilitates estimates of assay precision. Six calculation methods have been compared by evaluating the results of thousands of simulated, assays for three types of gamma-ray source distribution and ten masses. We will report on these computational studies and their experimental verification

  3. Pulmonary arterial lesions in explanted lungs after transplantation correlate with severity of pulmonary hypertension in chronic obstructive pulmonary disease

    DEFF Research Database (Denmark)

    Carlsen, Jørn; Andersen, Kasper Hasseriis; Boesgaard, Søren

    2013-01-01

    BACKGROUND: Pulmonary vascular findings are largely unreported in end-stage chronic obstructive pulmonary disease (COPD). METHODS: Pulmonary vascular lesions in explanted lungs from 70 patients with COPD/emphysema or α-1-antitrypsin deficiency were analyzed retrospectively. Patients were stratified...... of pulmonary vascular lesions in COPD correlate with the severity of PH. Morphologic lesions similar to those characteristic of IPAH can be observed as PH in COPD progresses to levels characteristic of IPAH....... by the presence and severity of pulmonary hypertension (PH) assessed by right-heart catheterization in 3 hemodynamically distinct groups: (1) non-PH (mean pulmonary arterial pressure [mPAP]50 mm Hg; median HE Grade 4 (range 3-6), with generalized arterial dilatation and plexiform lesions. CONCLUSIONS: The extent...

  4. Nicotine promotes rooting in leaf explants of in vitro raised seedlings of tomato, Lycopersicon esculentum Miller var. Pusa Ruby.

    Science.gov (United States)

    Bamel, Kiran; Gupta, Rajendra; Gupta, Shrish C

    2015-11-01

    Nicotine promotes rooting in leaf explants of tomato (Lycopersicon esculentum Miller var. Pusa Ruby). Nicotine at 10(-9) to 10(-3) M concentrations was added to the MS basal medium. The optimum response (three-fold increase in rooting) was obtained at 10(-7) M nicotine-enriched MS medium. At the same level i.e. 10(-7) M Nicotine induced dramatic increase (11-fold) in the number of secondary roots per root. We have shown earlier that exogenous acetylcholine induces a similar response in tomato leaves. Since nicotine is an agonist of one of the two acetylcholine receptors in animals, its ability to simulate ACh action in a plant system suggests the presence of the same molecular mechanism operative in both, animal and plant cells. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Nonparenchymal cells cultivated from explants of fibrotic liver resemble endothelial and smooth muscle cells from blood vessel walls

    International Nuclear Information System (INIS)

    Voss, B.; Rauterberg, J.; Pott, G.; Brehmer, U.; Allam, S.; Lehmann, R.; von Bassewitz, D.B.

    1982-01-01

    Tissue specimens from human fibrotic liver obtained by needle biopsy were cultured. Two cell types emerged from the tissue explants. From their morphology and biosynthetic products they resembled smooth muscle cells and endothelial cells from blood vessel walls. In the endothelial cells, factor VIII-associated protein was demonstrated by indirect immunofluorescence. Synthesis of collagen types I and III, basement membrane collagen types IV and V, and fibronectin by both cell types was observed by immunofluorescence microscopy. Homogeneous cultures of smooth muscle cells were observed in subcultures. After incubation with [ 14 C]glycine, collagen was isolated and characterized by CM cellulose chromatography, and consisted mainly of types I and III. These data suggest involvement of mesenchymal cells in hepatic fibrosis; they presumably originate from blood vessel or sinusoidal walls

  6. Enhanced unscheduled DNA synthesis in UV-irradiated human skin explants treated with T4N5 liposomes

    International Nuclear Information System (INIS)

    Yarosh, D.B.; Kibitel, J.T.; Green, L.A.; Spinowitz, A.

    1991-01-01

    Epidermal keratinocytes cultured from explants of skin cancer patients, including biopsies from xeroderma pigmentosum patients, were ultraviolet light-irradiated and DNA repair synthesis was measured. Repair capacity was much lower in xeroderma pigmentosum patients than in normal patients. The extent of DNA repair replication did not decline with the age of the normal patient. Treatment with T4N5 liposomes containing a DNA repair enzyme enhanced repair synthesis in both normal and xeroderma pigmentosum keratinocytes in an irradiation- and liposome-dose dependent manner. These results provide no evidence that aging people or skin cancer patients are predisposed to cutaneous malignancy by a DNA repair deficiency, but do demonstrate that T4N5 liposomes enhance DNA repair in the keratinocytes of the susceptible xeroderma pigmentosum and skin cancer population

  7. CT Fluoroscopy-Guided Lung Biopsy with Novel Steerable Biopsy Canula: Ex-Vivo Evaluation in Ventilated Porcine Lung Explants

    International Nuclear Information System (INIS)

    Schaefer, Philipp J.; Fabel, Michael; Bolte, Hendrik; Schaefer, Fritz K. W.; Jahnke, Thomas; Heller, Martin; Lammer, Johannes; Biederer, Juergen

    2010-01-01

    The purpose was to evaluate ex-vivo a prototype of a novel biopsy canula under CT fluoroscopy-guidance in ventilated porcine lung explants in respiratory motion simulations. Using an established chest phantom for porcine lung explants, n = 24 artificial lesions consisting of a fat-wax-Lipiodol mixture (approx. 70HU) were placed adjacent to sensible structures such as aorta, pericardium, diaphragm, bronchus and pulmonary artery. A piston pump connected to a reservoir beneath a flexible silicone reconstruction of a diaphragm simulated respiratory motion by rhythmic inflation and deflation of 1.5 L water. As biopsy device an 18-gauge prototype biopsy canula with a lancet-like, helically bended cutting edge was used. The artificial lesions were punctured under CT fluoroscopy-guidance (SOMATOM Sensation 64, Siemens, Erlangen, Germany; 30mAs/120 kV/5 mm slice thickness) implementing a dedicated protocol for CT fluoroscopy-guided lung biopsy. The mean-diameter of the artificial lesions was 8.3 ± 2.6 mm, and the mean-distance of the phantom wall to the lesions was 54.1 ± 13.5 mm. The mean-displacement of the lesions by respiratory motion was 14.1 ± 4.0 mm. The mean-duration of CT fluoroscopy was 9.6 ± 5.1 s. On a 4-point scale (1 = central; 2 = peripheral; 3 = marginal; 4 = off target), the mean-targeted precision was 1.9 ± 0.9. No misplacement of the biopsy canula affecting adjacent structures could be detected. The novel steerable biopsy canula proved to be efficient in the ex-vivo set-up. The chest phantom enabling respiratory motion and the steerable biopsy canula offer a feasible ex-vivo system for evaluating and training CT fluoroscopy-guided lung biopsy adapted to respiratory motion.

  8. A valued Indian medicinal plant – Begonia malabarica Lam. : Successful plant regeneration through various explants and field performance

    Directory of Open Access Journals (Sweden)

    Sevanan Rajeshkumar

    2009-05-01

    Full Text Available A cost-effective and efficient protocol has been described in the present work for large-scale and rapid in vitro propagation of a valuable medicinal herb Begonia malabarica Lam. (Begoniaceae by shoot auxillary-bud proliferation and organogenesis on MS medium supplemented with 6-benzylaminopurine (BA; 0.0-8.8 mg/l and indole-3-acetic acid (IAA; 0.0-2.88 mg/l at different concentrations, either alone or in combinations. Initiation of callus formation from the base of the leaf lamina was observed on MS supplemented with BA, IAA and adenine sulphate. Root induction on shoots was achieved on full strength MS with IAA/ indole-3-butyric acid (IBA at different concentrations. MS medium with 4.4 mg/l BA and 1.4 mg/l IAA elicited the maximum number of shoots (10 multiple shoots from nodal explants. Leaf-based callus differentiated into more than 28 shoots on MS with 150 mg/l adenine sulphate. The regenerated shoots were rooted on MS with 1.2 mg/l IBA within ten days. Almost 95% of the rooted shoots survived hardening when transferred to the field. The regenerated plants did not show any morphological change and variation in levels of secondary metabolites when compared with the mother stock. Thus, a reproduction of B. malabarica was established through nodal and leaf explants. This protocol can be exploited for conservation and commercial propagation of this medical plant in the Indian subcontinent and might be useful for genetic improvement programs.

  9. T-DNA transfer and T-DNA integration efficiencies upon Arabidopsis thaliana root explant cocultivation and floral dip transformation.

    Science.gov (United States)

    Ghedira, Rim; De Buck, Sylvie; Van Ex, Frédéric; Angenon, Geert; Depicker, Ann

    2013-12-01

    T-DNA transfer and integration frequencies during Agrobacterium-mediated root explant cocultivation and floral dip transformations of Arabidopsis thaliana were analyzed with and without selection for transformation-competent cells. Based on the presence or absence of CRE recombinase activity without or with the CRE T-DNA being integrated, transient expression versus stable transformation was differentiated. During root explant cocultivation, continuous light enhanced the number of plant cells competent for interaction with Agrobacterium and thus the number of transient gene expression events. However, in transformation competent plant cells, continuous light did not further enhance cotransfer or cointegration frequencies. Upon selection for root transformants expressing a first T-DNA, 43-69 % of these transformants showed cotransfer of another non-selected T-DNA in two different light regimes. However, integration of the non-selected cotransferred T-DNA occurred only in 19-46 % of these transformants, indicating that T-DNA integration in regenerating root cells limits the transformation frequencies. After floral dip transformation, transient T-DNA expression without integration could not be detected, while stable T-DNA transformation occurred in 0.5-1.3 % of the T1 seedlings. Upon selection for floral dip transformants with a first T-DNA, 8-34 % of the transformants showed cotransfer of the other non-selected T-DNA and in 93-100 % of them, the T-DNA was also integrated. Therefore, a productive interaction between the agrobacteria and the female gametophyte, rather than the T-DNA integration process, restricts the floral dip transformation frequencies.

  10. What are Segments in Google Analytics

    Science.gov (United States)

    Segments find all sessions that meet a specific condition. You can then apply this segment to any report in Google Analytics (GA). Segments are a way of identifying sessions and users while filters identify specific events, like pageviews.

  11. MICROPROPAGATION OF ADULT TREE OF PTEROCARPUS MARSUPIUM ROXB. USING NODAL EXPLANTS

    Directory of Open Access Journals (Sweden)

    Shipra JAISWAL

    2015-12-01

    Full Text Available Attempts were made for in vitro propagation of Pterocarpus marsupium Roxb., belonging to family Fabaceae, an economically important multipurpose tree. The tree is scared with noval antidiabetic properties. The tree shows poor seed germination capacity (30% due to hard seed coat and conventional vegetative regeneration methods are a complete failure. Therefore, the propagation of this tree by tissue culture techniques is an urgent need and well justified. Nodal segments containing axillary bud from 10 years old tree of P. marsupium were evaluated for axillary shoot proliferation on Murashige and Skoog’s (MS basal medium fortified with BAP (6–benzylaminopurine and kinetin (Kn singly or in combinations with auxins at different concentrations. The best shoot proliferation was obtained with 13.95 µM Kn + additives (568 µM Ascorbic acid, 260 µM Citric acid, 605 µM Ammonium sulphate and 217 µM Adenine sulphate in MS medium where 64.44% of the axillary buds responded with development of (2.51±0.10 shoots. Multiplication of in vitro shoots were achieved on MS Medium supplemented with Kn (9.30 µM + NAA (0.54 µM and additives. Half strength MS medium supplemented with 4.92 µM IBA induced in vitro rooting of in vitro shoots. In vitro regenerated plantlets with well developed roots were successfully hardened in a greenhouse.

  12. The release of bystander factor(s) from tissue explant cultures of rainbow trout (Onchorhynchus mykiss) after exposure to gamma radiation.

    Science.gov (United States)

    O'Dowd, Colm; Mothersill, Carmel E; Cairns, Michael T; Austin, Brian; McClean, Brendan; Lyng, Fiona M; Murphy, James E J

    2006-10-01

    The bystander response has been documented in cell lines and cell cultures derived from aquatic species over the past several years. However, little work has been undertaken to identify a similar bystander response in tissue explant cultures from fish. In this study, indirect effects of ionizing gamma radiation on tissue explant cultures of fish were investigated. Tissue explants in culture were exposed to 0.5 Gy and 5 Gy gamma radiation from a 60Co teletherapy unit. A bystander response in Epithelioma papulosum cyprini (EPC) cells exposed to gamma-irradiated tissue conditioned medium from rainbow trout explants was investigated, and the effects on cell survival were quantified by the clonogenic survival assay. Dichlorofluorescein and rhodamine 123 fluorescent dyes were used to identify alterations in reactive oxygen species (ROS) and mitochondrial membrane potential (MMP), respectively. Results indicate a different response for the three tissue types investigated. Clonogenic assay results vary from a decrease in cell survival (gill) to no effect (skin) to a stimulatory effect (spleen). Results from fluorescence assays of ROS and MMP show similarities to clonogenic assay results. This study identifies a useful model for further studies relating to the bystander effect in aquatic organisms in vivo and ex vivo.

  13. HSV-1 interaction to 3-O-sulfated heparan sulfate in mouse-derived DRG explant and profiles of inflammatory markers during virus infection

    NARCIS (Netherlands)

    Sharthiya, H.; Seng, C.; Kuppevelt, T.H. van; Tiwari, V.; Fornaro, M.

    2017-01-01

    The molecular mechanism of herpes simplex virus (HSV) entry and the associated inflammatory response in the nervous system remain poorly understood. Using mouse-derived ex vivo dorsal root ganglia (DRG) explant model and single cell neurons (SCNs), in this study, we provided a visual evidence for

  14. Reproductive survival of explanted human tumor cells after exposure to nitrogen mustard or x irradiation; differences in response with subsequent subculture in vitro

    International Nuclear Information System (INIS)

    Wells, J.; Berry, R.J.; Laing, A.H.

    1977-01-01

    Curves for the survival of reproductive capacity of explanted human tumor cells, following exposure to the alkylating agent nitrogen mustard (mustine hydrochloride) or 250-kVp x rays, were obtained as soon as a satisfactory plating efficiency, i.e., greater than or approximately equal to 10 percent, was obtained from the tumor cells in vitro (usually within 2-10 weeks of explanation). It was found that all six tumor explants tested became more sensitive to the action of nitrogen mustard on serial subculture, whereas the response of four explants which were X-irradiated was invariant with further subculturing. Furthermore, all but one explant yielded survival curves which were extremely similar, with D/sub q/ values circa 440-610 rad. One line, from a seminoma, however, had a D/sub q/ of 150 rad. These radiosensitive seminoma cells were, however, the most resistant to the action of nitrogen mustard. The increase in sensitivity to nitrogen mustard with serial subculture in vitro was not associated with any change in the proliferative rate of the cells, although it may be associated with an increase in the efficiency of transport

  15. CLG for Automatic Image Segmentation

    OpenAIRE

    Christo Ananth; S.Santhana Priya; S.Manisha; T.Ezhil Jothi; M.S.Ramasubhaeswari

    2017-01-01

    This paper proposes an automatic segmentation method which effectively combines Active Contour Model, Live Wire method and Graph Cut approach (CLG). The aim of Live wire method is to provide control to the user on segmentation process during execution. Active Contour Model provides a statistical model of object shape and appearance to a new image which are built during a training phase. In the graph cut technique, each pixel is represented as a node and the distance between those nodes is rep...

  16. Market segmentation, targeting and positioning

    OpenAIRE

    Camilleri, Mark Anthony

    2017-01-01

    Businesses may not be in a position to satisfy all of their customers, every time. It may prove difficult to meet the exact requirements of each individual customer. People do not have identical preferences, so rarely does one product completely satisfy everyone. Many companies may usually adopt a strategy that is known as target marketing. This strategy involves dividing the market into segments and developing products or services to these segments. A target marketing strategy is focused on ...

  17. Recognition Using Classification and Segmentation Scoring

    National Research Council Canada - National Science Library

    Kimball, Owen; Ostendorf, Mari; Rohlicek, Robin

    1992-01-01

    .... We describe an approach to connected word recognition that allows the use of segmental information through an explicit decomposition of the recognition criterion into classification and segmentation scoring...

  18. Somatic Embryogenesis and Plant Regeneration in Eggplant ...

    African Journals Online (AJOL)

    DR SIDHU

    2013-02-20

    Feb 20, 2013 ... Two as well as three way interactions of three eggplant genotypes, media compositions and explants. (hypocotyl, cotyledon and leaf) showed significant differences for plant regeneration. Among three explants, hypocotyl induced highest percent callusing, but cotyledon showed best results for somatic.

  19. Effect of explant density and volume of cultivation medium on in-vitro multiplication of blueberry (Vaccinium corymbosum L. varieties "Brigitta" and "Legacy"

    Directory of Open Access Journals (Sweden)

    Mario Rodríguez Beraud

    2015-03-01

    Full Text Available The objective of the investigation was to evaluate the in-vitro multiplication of two varieties of blueberry (Vaccinium corymbosum L., “Brigitta” and “Legacy” in response to five explants densities (5, 10, 15, 20 and 25 and four flask volumes (10, 20, 30 and 40 mL for cultivation. For both varieties the cultivation medium WPM (Woody Plant Medium was used. The experiment was completely randomized with 20 treatments and 12 repetitions per treatment. After 45 days of cultivation we evaluated the height of shoots, number of shoots/explant, number of nodes/shoot and number of shoots/flask. Variety “Brigitta” had highest shoots at higher densities and flask volumes, while variety “Legacy” had the highest average shoot height with intermediate densities and high volumes. Regarding the number of shoots/explant, the volume of the medium had no influence on “Brigitta”, however, higher plant densities affected this parameter. With variety “Legacy” the maximum number of shoots was achieved with lower plant densities and intermediate culture volumes per flask. In relation to the number of nodes per explant "Brigitta had lower numbers as compared to “Legacy”, but with both varieties the number of nodes decresed with smaller volumes of medium in the flasks. For the number of shoots per flask, “Brigitta” responsed best at higher densities exceeding 40 shoots per flask. In contrast, “Legacy” produced maximum results at density of 25 explants in 30 mL of medium. It is concluded that for the optimum multiplication of both varieties the correct selection of both, the planting density and the volume of multiplication medium are important.

  20. Changes in the metabolic footprint of placental explant-conditioned medium cultured in different oxygen tensions from placentas of small for gestational age and normal pregnancies.

    LENUS (Irish Health Repository)

    Horgan, R P

    2012-01-31

    Being born small for gestational age (SGA) confers significantly increased risks of perinatal morbidity and mortality. Accumulating evidence suggests that an SGA fetus results from a poorly perfused and abnormally developed placenta. Some of the placental features seen in SGA, such as abnormal cell turnover and impaired nutrient transport, can be reproduced by culture of placental explants in hypoxic conditions. Metabolic footprinting offers a hypothesis-generating strategy to investigate factors absorbed by and released from this tissue in vitro. Previously, metabolic footprinting of the conditioned culture media has identified differences in placental explants cultured under normoxic and hypoxic conditions and between normal pregnancies and those complicated by pre-eclampsia. In this study we aimed to examine the differences in the metabolic footprint of placental villous explants cultured at different oxygen (O(2)) tensions between women who deliver an SGA baby (n = 9) and those from normal controls (n = 8). Placental villous explants from cases and controls were cultured for 96 h in 1% (hypoxic), 6% (normoxic) and 20% (hyperoxic) O(2). Metabolic footprints were analysed by Ultra Performance Liquid Chromatography coupled to an electrospray hybrid LTQ-Orbitrap Mass Spectrometry (UPLC-MS). 574 metabolite features showed significant difference between SGA and normal at one or more of the oxygen tensions. SGA explant media cultured under hypoxic conditions was observed, on a univariate level, to exhibit the same metabolic signature as controls cultured under normoxic conditions in 49% of the metabolites of interest, suggesting that SGA tissue is acclimatised to hypoxic conditions in vivo. No such behaviour was observed under hyperoxic culture conditions. Glycerophospholipid and tryptophan metabolism were highlighted as areas of particular interest.

  1. Methods of evaluating segmentation characteristics and segmentation of major faults

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Kie Hwa; Chang, Tae Woo; Kyung, Jai Bok [Seoul National Univ., Seoul (Korea, Republic of)] (and others)

    2000-03-15

    Seismological, geological, and geophysical studies were made for reasonable segmentation of the Ulsan fault and the results are as follows. One- and two- dimensional electrical surveys revealed clearly the fault fracture zone enlarges systematically northward and southward from the vicinity of Mohwa-ri, indicating Mohwa-ri is at the seismic segment boundary. Field Geological survey and microscope observation of fault gouge indicates that the Quaternary faults in the area are reactivated products of the preexisting faults. Trench survey of the Chonbuk fault Galgok-ri revealed thrust faults and cumulative vertical displacement due to faulting during the late Quaternary with about 1.1-1.9 m displacement per event; the latest event occurred from 14000 to 25000 yrs. BP. The seismic survey showed the basement surface os cut by numerous reverse faults and indicated the possibility that the boundary between Kyeongsangbukdo and Kyeongsannamdo may be segment boundary.

  2. Segmentation by Large Scale Hypothesis Testing - Segmentation as Outlier Detection

    DEFF Research Database (Denmark)

    Darkner, Sune; Dahl, Anders Lindbjerg; Larsen, Rasmus

    2010-01-01

    a microscope and we show how the method can handle transparent particles with significant glare point. The method generalizes to other problems. THis is illustrated by applying the method to camera calibration images and MRI of the midsagittal plane for gray and white matter separation and segmentation......We propose a novel and efficient way of performing local image segmentation. For many applications a threshold of pixel intensities is sufficient but determine the appropriate threshold value can be difficult. In cases with large global intensity variation the threshold value has to be adapted...... locally. We propose a method based on large scale hypothesis testing with a consistent method for selecting an appropriate threshold for the given data. By estimating the background distribution we characterize the segment of interest as a set of outliers with a certain probability based on the estimated...

  3. Methods of evaluating segmentation characteristics and segmentation of major faults

    International Nuclear Information System (INIS)

    Lee, Kie Hwa; Chang, Tae Woo; Kyung, Jai Bok

    2000-03-01

    Seismological, geological, and geophysical studies were made for reasonable segmentation of the Ulsan fault and the results are as follows. One- and two- dimensional electrical surveys revealed clearly the fault fracture zone enlarges systematically northward and southward from the vicinity of Mohwa-ri, indicating Mohwa-ri is at the seismic segment boundary. Field Geological survey and microscope observation of fault gouge indicates that the Quaternary faults in the area are reactivated products of the preexisting faults. Trench survey of the Chonbuk fault Galgok-ri revealed thrust faults and cumulative vertical displacement due to faulting during the late Quaternary with about 1.1-1.9 m displacement per event; the latest event occurred from 14000 to 25000 yrs. BP. The seismic survey showed the basement surface os cut by numerous reverse faults and indicated the possibility that the boundary between Kyeongsangbukdo and Kyeongsannamdo may be segment boundary

  4. Automatic segmentation of psoriasis lesions

    Science.gov (United States)

    Ning, Yang; Shi, Chenbo; Wang, Li; Shu, Chang

    2014-10-01

    The automatic segmentation of psoriatic lesions is widely researched these years. It is an important step in Computer-aid methods of calculating PASI for estimation of lesions. Currently those algorithms can only handle single erythema or only deal with scaling segmentation. In practice, scaling and erythema are often mixed together. In order to get the segmentation of lesions area - this paper proposes an algorithm based on Random forests with color and texture features. The algorithm has three steps. The first step, the polarized light is applied based on the skin's Tyndall-effect in the imaging to eliminate the reflection and Lab color space are used for fitting the human perception. The second step, sliding window and its sub windows are used to get textural feature and color feature. In this step, a feature of image roughness has been defined, so that scaling can be easily separated from normal skin. In the end, Random forests will be used to ensure the generalization ability of the algorithm. This algorithm can give reliable segmentation results even the image has different lighting conditions, skin types. In the data set offered by Union Hospital, more than 90% images can be segmented accurately.

  5. Skip segment Hirschsprung disease and Waardenburg syndrome

    Directory of Open Access Journals (Sweden)

    Erica R. Gross

    2015-04-01

    Full Text Available Skip segment Hirschsprung disease describes a segment of ganglionated bowel between two segments of aganglionated bowel. It is a rare phenomenon that is difficult to diagnose. We describe a recent case of skip segment Hirschsprung disease in a neonate with a family history of Waardenburg syndrome and the genetic profile that was identified.

  6. U.S. Army Custom Segmentation System

    Science.gov (United States)

    2007-06-01

    segmentation is individual or intergroup differences in response to marketing - mix variables. Presumptions about segments: •different demands in a...product or service category, •respond differently to changes in the marketing mix Criteria for segments: •The segments must exist in the environment

  7. Skip segment Hirschsprung disease and Waardenburg syndrome

    OpenAIRE

    Gross, Erica R.; Geddes, Gabrielle C.; McCarrier, Julie A.; Jarzembowski, Jason A.; Arca, Marjorie J.

    2015-01-01

    Skip segment Hirschsprung disease describes a segment of ganglionated bowel between two segments of aganglionated bowel. It is a rare phenomenon that is difficult to diagnose. We describe a recent case of skip segment Hirschsprung disease in a neonate with a family history of Waardenburg syndrome and the genetic profile that was identified.

  8. A Novel Iris Segmentation Scheme

    Directory of Open Access Journals (Sweden)

    Chen-Chung Liu

    2014-01-01

    Full Text Available One of the key steps in the iris recognition system is the accurate iris segmentation from its surrounding noises including pupil, sclera, eyelashes, and eyebrows of a captured eye-image. This paper presents a novel iris segmentation scheme which utilizes the orientation matching transform to outline the outer and inner iris boundaries initially. It then employs Delogne-Kåsa circle fitting (instead of the traditional Hough transform to further eliminate the outlier points to extract a more precise iris area from an eye-image. In the extracted iris region, the proposed scheme further utilizes the differences in the intensity and positional characteristics of the iris, eyelid, and eyelashes to detect and delete these noises. The scheme is then applied on iris image database, UBIRIS.v1. The experimental results show that the presented scheme provides a more effective and efficient iris segmentation than other conventional methods.

  9. Rod and cone photoreceptor cells produce ROS in response to stress in a live retinal explant system.

    LENUS (Irish Health Repository)

    Bhatt, Lavinia

    2010-01-01

    PURPOSE: The production of reactive oxygen species (ROS) can lead to oxidative stress, which is a strong contributory factor to many ocular diseases. In this study, the removal of trophic factors is used as a model system to investigate the effects of stress in the retina. The aims were to determine if both rod and cone photoreceptor cells produce ROS when they are deprived of trophic factor support and to demonstrate if the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (Nox) enzymes are responsible for this ROS production. METHODS: Retinas were explanted from mice aged between postnatal days 8-10 and cultured overnight. The following morning, confocal microscopy combined with various fluorescent probes was used to detect the production of ROS. Each time peanut agglutinin (PNA), a cone photoreceptor marker, was used to facilitate orientation of the retina. Dihydroethidium and dihydrorhodamine 123 (DHR123) were used to determine which cells produce ROS. Subsequently, western blots of retinal serial sections were used to detect the presence of Noxs in the different retinal layers. The Nox inhibitor apocynin was then tested to determine if it altered the production of ROS within these cells. RESULTS: Live retinal explants, viewed at high magnifications using confocal microscopy, displayed an increase in the fluorescent products of dihydroethidium and DHR123 upon serum removal when compared to controls. DHR123 fluorescence, once oxidized, localized to mitochondria and was found in the same focal plane as the PNA staining. This showed that cones and rods produced ROS when stressed. Retinal serial sectioning established that the photoreceptor layer expressed Nox4, dual oxidase (Duox) 1, and Duox2 at varying levels. Finally, the Nox inhibitor apocynin decreased the burst stimulated by the stress of serum removal. CONCLUSIONS: Confocal microscopy and PNA staining allowed differentiation of cell types within the outermost layers of the retina, demonstrating

  10. Notochord manipulation does not impact oesophageal and tracheal formation from isolated foregut in 3D explant culture.

    Science.gov (United States)

    Mc Laughlin, Danielle; Murphy, Paula; Puri, Prem

    2016-01-01

    Tracheo-oesophageal malformations result from disturbed foregut separation during early development. The notochord, a specialised embryonic structure, forms immediately adjacent to the dividing foregut. In the Adriamycin mouse model of oesophageal atresia, foregut and notochord abnormalities co-exist, and the site and severity of foregut malformations closely correlate to the position and extent of the notochord defects. Notochord and foregut abnormalities also co-exist in the Noggin Knockout mouse as well in a small number of human cases. The notochord is a source of powerful molecular signals during early embryogenesis, being particularly important for neural crest development. The influence of notochord signaling on the adjacent foregut is not known. The purpose of this study was to examine the impact of notochord manipulation on foregut separation using a robust 3D explant method for culturing isolated foregut which permits oeosphageal and tracheal formation in vitro. Foregut was micro-dissected from embryonic day 9 mice (License B100/4447 Irish Medicines Board), embedded in collagen and cultured for 48 h with native notochord intact (n = 6), notochord removed (n = 10) or additional notochord transplanted from stage matched controls (n = 8). Specimens were analysed for foregut morphology and molecular patterning using immunohistochemistry for Hnf3b (an endoderm marker) and Sox2 (a notochord and oesophageal marker) on cryosections. Foregut separation into distinct oesophagus and trachea was observed in isolated foregut specimens with or without their native notochord. In specimens with additional notochord transplants, foregut morphology and molecular patterning were comparable to controls whether or not the native notochord was maintained. In particular foregut separation was not disrupted by the transplantation of additional notochord at the dorsal foregut endoderm. The relationship between the embryonic foregut and notochord is complex and ill

  11. Document segmentation via oblique cuts

    Science.gov (United States)

    Svendsen, Jeremy; Branzan-Albu, Alexandra

    2013-01-01

    This paper presents a novel solution for the layout segmentation of graphical elements in Business Intelligence documents. We propose a generalization of the recursive X-Y cut algorithm, which allows for cutting along arbitrary oblique directions. An intermediate processing step consisting of line and solid region removal is also necessary due to presence of decorative elements. The output of the proposed segmentation is a hierarchical structure which allows for the identification of primitives in pie and bar charts. The algorithm was tested on a database composed of charts from business documents. Results are very promising.

  12. Optimally segmented permanent magnet structures

    DEFF Research Database (Denmark)

    Insinga, Andrea Roberto; Bjørk, Rasmus; Smith, Anders

    2016-01-01

    We present an optimization approach which can be employed to calculate the globally optimal segmentation of a two-dimensional magnetic system into uniformly magnetized pieces. For each segment the algorithm calculates the optimal shape and the optimal direction of the remanent flux density vector......, with respect to a linear objective functional. We illustrate the approach with results for magnet design problems from different areas, such as a permanent magnet electric motor, a beam focusing quadrupole magnet for particle accelerators and a rotary device for magnetic refrigeration....

  13. In vitro regeneration of cocona (Solanum sessiliflorum, Solanaceae) cultivars for commercial production.

    Science.gov (United States)

    Schuelter, A R; Grunvald, A K; Amaral, A T; da Luz, C L; Luz, C L; Gonçalves, L M; Stefanello, S; Scapim, C A

    2009-08-11

    Cocona (Solanum sessiliflorum Dunal) is a solanaceous shrub native to the Amazon region that produces an edible fruit. This species has numerous advantages, particularly a high nutritional value and productivity. However, due to irregular germination and rapid loss of seed viability, there are few plantations for production on a large scale. Development of alternative propagation strategies is essential for the production of homogeneous seedlings of genotypes with superior agronomic performance. We developed techniques for in vitro regeneration of the cocona varieties Santa Luzia and Thaís for large-scale production of healthy plantlets. Twenty days after seeding, seedling segments germinated in vitro were used as explant sources. Three successive experiments were performed: one to test the effect of the explant source and combinations of two growth regulators, auxin (indole acetic acid, IAA) and kinetin (KIN), on the morphogenetic response; another to investigate the effect of the combination of growth regulators on the morphogenetic response of hypocotyl segments, and another to evaluate how sucrose concentration affects the development of adventitious shoots. The best shoot induction was obtained using hypocotyl segments and stem apices, while rhizogenesis was greatest in leaves with a petiole. The number of adventitious shoots per explant on hypocotyl segments increased with 10 and 20 mg/L KIN, combined with 0.02 mg/L IAA in the variety Santa Luzia. Sucrose combined with these growth regulator levels increased the average number of calli; these were optimally produced when 45 g/L sucrose and 0.01 mg/L IAA + 20 mg/L KIN were applied. Only sucrose concentration influenced shoot proliferation in the two S. sessiliflorum varieties, with a maximum at 17.5 g/L.

  14. Intercalary bone segment transport in treatment of segmental tibial defects

    International Nuclear Information System (INIS)

    Iqbal, A.; Amin, M.S.

    2002-01-01

    Objective: To evaluate the results and complications of intercalary bone segment transport in the treatment of segmental tibial defects. Design: This is a retrospective analysis of patients with segmental tibial defects who were treated with intercalary bone segment transport method. Place and Duration of Study: The study was carried out at Combined Military Hospital, Rawalpindi from September 1997 to April 2001. Subjects and methods: Thirteen patients were included in the study who had developed tibial defects either due to open fractures with bone loss or subsequent to bone debridement of infected non unions. The mean bone defect was 6.4 cms and there were eight associated soft tissue defects. Locally made unilateral 'Naseer-Awais' (NA) fixator was used for bone segment transport. The distraction was done at the rate of 1mm/day after 7-10 days of osteotomy. The patients were followed-up fortnightly during distraction and monthly thereafter. The mean follow-up duration was 18 months. Results: The mean time in external fixation was 9.4 months. The m ean healing index' was 1.47 months/cm. Satisfactory union was achieved in all cases. Six cases (46.2%) required bone grafting at target site and in one of them grafting was required at the level of regeneration as well. All the wounds healed well with no residual infection. There was no residual leg length discrepancy of more than 20 mm nd one angular deformity of more than 5 degrees. The commonest complication encountered was pin track infection seen in 38% of Shanz Screws applied. Loosening occurred in 6.8% of Shanz screws, requiring re-adjustment. Ankle joint contracture with equinus deformity and peroneal nerve paresis occurred in one case each. The functional results were graded as 'good' in seven, 'fair' in four, and 'poor' in two patients. Overall, thirteen patients had 31 (minor/major) complications with a ratio of 2.38 complications per patient. To treat the bone defects and associated complications, a mean of

  15. Indução e cultivo in vitro de gemas adventícias em segmentos de epicótilo de laranja-azeda In vitro induction and culture of adventitious buds in epicotyl segments of sour orange

    Directory of Open Access Journals (Sweden)

    Rosely Pereira da Silva

    2008-10-01

    Full Text Available O objetivo deste trabalho foi avaliar a indução e a formação de gemas adventícias em explantes de laranja-azeda, pelo uso de fitorreguladores. Em experimentos de organogênese in vitro foram avaliados 6-benzilaminopurina (BAP, thidiazuron (TDZ e cinetina (CIN, em diferentes concentrações e sob duas condições de luminosidade; BAP e CIN combinados ou não com ácido naftalenoacético (ANA; e BAP e CIN isoladamente ou combinados entre si. Segmentos de epicótilo de 1 cm de comprimento, provenientes de plântulas de laranja-azeda germinadas in vitro, foram utilizados como explantes. Para induzir a formação de gemas, os segmentos foram cultivados em meio MT com ou sem adição de fitorreguladores. O material foi cultivado a 27ºC em ausência de luz por 30 dias, seguidos de fotoperíodo de 16 horas. O delineamento experimental foi inteiramente casualizado, com quatro ou cinco repetições, a depender do experimento e, cada repetição foi constituída de placa de Petri com 20 explantes. Após 60 ou 70 dias de cultivo foram avaliados o percentual de explantes responsivos e o número de gemas por explante. A adição de BAP ao meio de cultura, combinada ou não com ANA, e em combinações com CIN promovem melhor resposta organogênica.The objective of this work was to evaluate the induction and formation of adventitious buds in sour orange explants through the use of plant regulators. In vitro organogenesis experiments were conducted to evaluate the effect of BAP, TDZ, and KIN in different concentrations and under two light conditions; BAP and KIN, combined or not with NAA; BAP and KIN, separately or in combined concentrations. Sour orange epicotyl segments (1 cm length, from in vitro germinated plants, were used as explants. In order to induce bud formation, the explants were cultured in MT medium with or without the addition of plant regulators. The material was cultivated at 27ºC in the absence of light for 30 days, followed of culture

  16. Hydrophilic segmented block copolymers based on poly(ethylene oxide) and monodisperse amide segments

    NARCIS (Netherlands)

    Husken, D.; Feijen, Jan; Gaymans, R.J.

    2007-01-01

    Segmented block copolymers based on poly(ethylene oxide) (PEO) flexible segments and monodisperse crystallizable bisester tetra-amide segments were made via a polycondensation reaction. The molecular weight of the PEO segments varied from 600 to 4600 g/mol and a bisester tetra-amide segment (T6T6T)

  17. Dose-dependency and reversibility of radiation-induced injury in cardiac explant-derived cells of mice

    Science.gov (United States)

    Luo, Lan; Yan, Chen; Urata, Yoshishige; Hasan, Al Shaimaa; Goto, Shinji; Guo, Chang-Ying; Zhang, Shouhua; Li, Tao-Sheng

    2017-01-01

    We evaluated the dose-dependency and reversibility of radiation-induced injury in cardiac explant-derived cells (CDCs), a mixed cell population grown from heart tissues. Adult C57BL/6 mice were exposed to 0, 10, 50 and 250 mGy γ-rays for 7 days and atrial tissues were collected for experiments 24 hours after last exposure. The number of CDCs was significantly decreased by daily exposure to over 250 mGy. Interestingly, daily exposure to over 50 mGy significantly decreased the c-kit expression and telomerase activity, increased 53BP1 foci in the nuclei of CDCs. However, CD90 expression and growth factors production in CDCs were not significantly changed even after daily exposure to 250 mGy. We further evaluated the reversibility of radiation-induced injury in CDCs at 1 week and 3 weeks after a single exposure to 3 Gy γ-rays. The number and growth factors production of CDCs were soon recovered at 1 week. However, the increased expression of CD90 were retained at 1 week, but recovered at 3 weeks. Moreover, the decreased expression of c-kit, impaired telomerase activity, and increased 53BP1 foci were poorly recovered even at 3 weeks. These data may help us to find the most sensitive and reliable bio-parameter(s) for evaluating radiation-induced injury in CDCs. PMID:28098222

  18. The effect of radiation in combination with carcinogens on the growth of normal urothelium in explant culture

    International Nuclear Information System (INIS)

    Mothersill, C.; O'Brien, A.

    1990-01-01

    Radiation is known to be carcinogenic to humans but attempts to demonstrate the process using human tissue culture models have met with little success. In the present study explants were established from urothelium and exposed to radiation and a range of chemical carcinogens, suspected promotor or metabolic agents. The resulting outgrowth was monitored for growth rate, proliferating epithelial fraction and development and differentiation of endothelial cells in culture. The results indicate that enhanced growth of epithelial cells can be seen when cultures are irradiated in the presence of various nitrosamines, benzo(a)pyrene or aniline. Radiation alone reduced the overall growth area measured but several proliferative foci developed on the resulting outgrowth. Their ultrastructural appearance reveals that they carry severe mitochondrial damage and exposure of treated cultures to metabolic inhibitors confirms that their respiration is defective. Endothelial cells proliferated over the surface of the epithelial monolayer and both the number and the degree of differentiation of the endothelial cells increased with increasing dose up to 10 Gy. While the cultures are not immortalised by the treatment, it appears that the epithelial cells have an extended lifespan (division capacity) and that a subpopulation has undergone a number of premalignant changes. Changes in endothelial cell proliferation also occur. (orig.)

  19. Accuracy of hepatocellular carcinoma detection on multidetector CT in a transplant liver population with explant liver correlation

    International Nuclear Information System (INIS)

    Addley, H.C.; Griffin, N.; Shaw, A.S.; Mannelli, L.; Parker, R.A.; Aitken, S.; Wood, H.; Davies, S.; Alexander, G.J.; Lomas, D.J.

    2011-01-01

    Aim: To evaluate the diagnostic accuracy of multidetector computed tomography (MDCT) for hepatocellular carcinoma (HCC) in cirrhotic patients undergoing liver transplantation. Secondary aims were to examine the effect of radiologist experience and lesion size on diagnostic accuracy. Materials and methods: Thirty-nine patients (72% male with a mean age of 56.5 years) underwent liver transplantation following preoperative triple-phase MDCT examination of the liver. MDCT examinations were retrospectively independently reviewed by three radiologists for the presence and location of suspected HCCs, with the diagnostic confidence recorded using a five-point confidence scale. MDCT examinations were compared with explant specimens for histopathological correlation. Results: Histopathological results demonstrated 46 HCCs in 29 of the 39 patients. Analysis demonstrated a sensitivity of 65-75% and specificity of 47-88% for detection of HCC lesions. The sensitivity dropped to 48-57% for lesions of size ≤20 mm. As the diagnostic confidence increased, there was a further decrease in the sensitivity (4-26%). The radiologist with the greatest number of years experience was found to have a significantly higher accuracy of detection of HCC lesions compared with the least experienced radiologist. Conclusion: Larger lesion size of HCC and greater number of years experience of the radiologist resulted in significantly higher accuracy of HCC lesion detection. The overall sensitivity and specificity results for MDCT detection of HCC are comparable to previous helical CT imaging.

  20. Experimental analysis of control mechanisms in somite segmentation in avian embryos. II. Reduction of material in the gastrula stages of the chick.

    Science.gov (United States)

    Bellairs, R; Veini, M

    1984-02-01

    A new theory of control of somite segmentation in chick embryos is proposed. This supposses that tiny clusters of already programmed cells are present throughout the presumptive somite area at stage 4, but that in order to fulfill their destiny they probably depend on the addition of further cells from the primitive streak. Evidence is based on the two groups of experiments: a) Experiments involving transection across the primitive streak at various stages, (which results in a 'tail' which possesses mesodermal derivatives) and across the segmental plate (which results in a 'tail' lacking mesodermal derivatives). b) Experiments in which parts of embryos have been explanted with or without their primitive streak. It is suggested that the initial clusters of pre-programmed cells move further and further posteriorly, developing into somitomeres (the precursors of true somites) only as they receive re-inforcements from the primitive streak or, ultimately, from the tail bud.

  1. Dictionary Based Segmentation in Volumes

    DEFF Research Database (Denmark)

    Emerson, Monica Jane; Jespersen, Kristine Munk; Jørgensen, Peter Stanley

    Method for supervised segmentation of volumetric data. The method is trained from manual annotations, and these annotations make the method very flexible, which we demonstrate in our experiments. Our method infers label information locally by matching the pattern in a neighborhood around a voxel ...... to a dictionary, and hereby accounts for the volume texture....

  2. Multiple Segmentation of Image Stacks

    DEFF Research Database (Denmark)

    Smets, Jonathan; Jaeger, Manfred

    2014-01-01

    We propose a method for the simultaneous construction of multiple image segmentations by combining a recently proposed “convolution of mixtures of Gaussians” model with a multi-layer hidden Markov random field structure. The resulting method constructs for a single image several, alternative...

  3. Segmenting Trajectories by Movement States

    NARCIS (Netherlands)

    Buchin, M.; Kruckenberg, H.; Kölzsch, A.; Timpf, S.; Laube, P.

    2013-01-01

    Dividing movement trajectories according to different movement states of animals has become a challenge in movement ecology, as well as in algorithm development. In this study, we revisit and extend a framework for trajectory segmentation based on spatio-temporal criteria for this purpose. We adapt

  4. Segmental Colitis Complicating Diverticular Disease

    Directory of Open Access Journals (Sweden)

    Guido Ma Van Rosendaal

    1996-01-01

    Full Text Available Two cases of idiopathic colitis affecting the sigmoid colon in elderly patients with underlying diverticulosis are presented. Segmental resection has permitted close review of the histopathology in this syndrome which demonstrates considerable similarity to changes seen in idiopathic ulcerative colitis. The reported experience with this syndrome and its clinical features are reviewed.

  5. Leaf segmentation in plant phenotyping

    NARCIS (Netherlands)

    Scharr, Hanno; Minervini, Massimo; French, Andrew P.; Klukas, Christian; Kramer, David M.; Liu, Xiaoming; Luengo, Imanol; Pape, Jean Michel; Polder, Gerrit; Vukadinovic, Danijela; Yin, Xi; Tsaftaris, Sotirios A.

    2016-01-01

    Image-based plant phenotyping is a growing application area of computer vision in agriculture. A key task is the segmentation of all individual leaves in images. Here we focus on the most common rosette model plants, Arabidopsis and young tobacco. Although leaves do share appearance and shape

  6. Acetylcholine suppresses shoot formation and callusing in leaf explants of in vitro raised seedlings of tomato, Lycopersicon esculentum Miller var. Pusa Ruby.

    Science.gov (United States)

    Bamel, Kiran; Gupta, Rajendra; Gupta, Shirish C

    2016-06-02

    We present experimental evidence to show that acetylcholine (ACh) causes decrease in shoot formation in leaf explants of tomato (Lycopersicon esculentum Miller var Pusa Ruby) when cultured on shoot regeneration medium. The optimum response was obtained at 10(-4) M ACh-enriched medium. ACh also causes decrease in percentage of cultures forming callus and reduces the callus mass. Inhibitors of enzymatic hydrolysis of ACh, neostigmine and physostigmine, also suppresses callogenesis and caulogenesis. On the other hand, the breakdown products of Ach, choline and acetate, do not alter the morphogenic response induced on the shoot regeneration medium. Neostigmine showed optimal reduction in shoot formation at 10(-5) M. The explants cultured on neostigmine augmented medium showed decline in the activity of ACh hydrolyzing enzyme acetylcholinesterase. ACh and neostigmine added together showed marked reduction in callus mass. These results strongly support the role of ACh as a natural regulator of morphogenesis in tomato plants.

  7. Acetylcholine suppresses shoot formation and callusing in leaf explants of in vitro raised seedlings of tomato, Lycopersicon esculentum Miller var. Pusa Ruby

    Science.gov (United States)

    Bamel, Kiran; Gupta, Rajendra; Gupta, Shirish C.

    2016-01-01

    ABSTRACT We present experimental evidence to show that acetylcholine (ACh) causes decrease in shoot formation in leaf explants of tomato (Lycopersicon esculentum Miller var Pusa Ruby) when cultured on shoot regeneration medium. The optimum response was obtained at 10−4 M ACh-enriched medium. ACh also causes decrease in percentage of cultures forming callus and reduces the callus mass. Inhibitors of enzymatic hydrolysis of ACh, neostigmine and physostigmine, also suppresses callogenesis and caulogenesis. On the other hand, the breakdown products of Ach, choline and acetate, do not alter the morphogenic response induced on the shoot regeneration medium. Neostigmine showed optimal reduction in shoot formation at 10−5 M. The explants cultured on neostigmine augmented medium showed decline in the activity of ACh hydrolyzing enzyme acetylcholinesterase. ACh and neostigmine added together showed marked reduction in callus mass. These results strongly support the role of ACh as a natural regulator of morphogenesis in tomato plants. PMID:27348536

  8. Pars plana Ahmed valve and vitrectomy in patients with glaucoma associated with posterior segment disease.

    Science.gov (United States)

    Wallsh, Josh O; Gallemore, Ron P; Taban, Mehran; Hu, Charles; Sharareh, Behnam

    2013-01-01

    To assess the safety and efficacy of a modified technique for pars plana placement of the Ahmed valve in combination with pars plana vitrectomy in the treatment of glaucoma associated with posterior segment disease. Thirty-nine eyes with glaucoma associated with posterior segment disease underwent pars plana vitrectomy combined with Ahmed valve placement. All valves were placed in the pars plana using a modified technique, without the pars plana clip, and using a scleral patch graft. The 24 eyes diagnosed with neovascular glaucoma had an improvement in intraocular pressure from 37.6 mmHg to 13.8 mmHg and best-corrected visual acuity from 2.13 logarithm of minimum angle of resolution to 1.40 logarithm of minimum angle of resolution. Fifteen eyes diagnosed with steroid-induced glaucoma had an improvement in intraocular pressure from 27.9 mmHg to 14.1 mmHg and best-corrected visual acuity from 1.38 logarithm of minimum angle of resolution to 1.13 logarithm of minimum angle of resolution. Complications included four cases of cystic bleb formation and one case of choroidal detachment and explantation for hypotony. Ahmed valve placement through the pars plana during vitrectomy is an effective option for managing complex cases of glaucoma without the use of the pars plana clip.

  9. Joint shape segmentation with linear programming

    KAUST Repository

    Huang, Qixing

    2011-01-01

    We present an approach to segmenting shapes in a heterogenous shape database. Our approach segments the shapes jointly, utilizing features from multiple shapes to improve the segmentation of each. The approach is entirely unsupervised and is based on an integer quadratic programming formulation of the joint segmentation problem. The program optimizes over possible segmentations of individual shapes as well as over possible correspondences between segments from multiple shapes. The integer quadratic program is solved via a linear programming relaxation, using a block coordinate descent procedure that makes the optimization feasible for large databases. We evaluate the presented approach on the Princeton segmentation benchmark and show that joint shape segmentation significantly outperforms single-shape segmentation techniques. © 2011 ACM.

  10. INTRASTROMAL CORNEAL RING SEGMENTS (ICRS, KERAVISION RING, INTACSTM: CLINICAL RESULTS AFTER 2 YEARS

    Directory of Open Access Journals (Sweden)

    Josef Ruckhofer

    2002-12-01

    Full Text Available Background. Since 1996 Intrastromal Corneal Ring Segments (IntacsTM, KeraVision, Inc.Fremont, Ca, USA have been used for the correction of mild to moderate myopia at the Salzburg Eye Clinic. Aim of this study was to evaluate the stability, reversibility and adjustability for this new method.Patients and methods. Our experience and final results of 54 surgeries – 30 eyes with a minimum follow-up of 2 years – as well as the potential reversibility (3 explantations, 2 of them followed by PRK and adjustability (3 exchanges are reported in detail.Results. None of the eyes lost more than one line in BSCVA (in 14 of 30 eyes the BSCVA improved. 73% (22/30 eyes reached an UCVA of 1.0 or better, 47% (14/30 eyes of 1.25 or better. After 2 years 47% (14/30 eyes were within ± 0.5 Dsph of the attempted correction. We observed no significant intraor postoperative complications. After ICRS removal the refractive data returned within ± 0.75 Dsph (MRSE and ± 0.5 Dsph (mean keratometry of preoperative values, respectively. Patients with an ICRS exchange obtained an improved visual acuity between 0.8 and 1.0, gaining between 2 to 4 lines.Conclusions. With the follow-up period of 2 years the ICRS seem to provide a very stable correction of low to moderate myopia. The procedure is reversible to a large extent, potentially adjustable (within certain limits and carries a minimal risk only. After explantation PRK can be performed with good visual results.

  11. Improvement in Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) by the inhibition of polyphenolics released during wounding of cotyledonary node explants.

    Science.gov (United States)

    Yadav, Reena; Mehrotra, Meenakshi; Singh, Aditya K; Niranjan, Abhishek; Singh, Rani; Sanyal, Indraneel; Lehri, Alok; Pande, Veena; Amla, D V

    2017-01-01

    Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) has been performed using cotyledonary node explants (CNs), which release phenolics upon excision that are detrimental to the viability of Agrobacterium tumefaciens and result in low transformation frequency. Twelve low molecular weight phenolic compounds and salicylic acid were identified in the exudates released upon excision during the preparation of cotyledonary nodes by reverse phase high-performance liquid chromatography (RP-HPLC). Zone inhibition assays performed with the explant exudates released at periodic intervals after excision showed the inhibition of A. tumefaciens. Agroinoculation of freshly excised cotyledonary nodes of chickpea showed 98-99 % inhibition of colony forming units (cfu). Osmium tetraoxide fixation of excised tissues showed enhanced accumulation of phenolics in the sub-epidermal regions causing enzymatic browning, affecting the viability and performance of A. tumefaciens for T-DNA delivery. The periodic analysis of exudates released from excised CNs showed enhanced levels of gallic acid (0.2945 ± 0.014 μg/g), chlorogenic acid (0.0978 ± 0.0046 μg/g), and quercetin (0.0971 ± 0.0046 μg/g) fresh weight, which were detrimental to A. tumefaciens. Quantitative assays and the elution profile showed the maximum leaching of phenolics, flavonoids, and salicylic acid immediately after the excision of explants and continued till 4 to 8 h post-excision. Pre-treatment of excised explants with inhibitors of polyphenol oxidase like L-cysteine, DTT, and sodium thiosulfate before co-cultivation showed the recovery of A. tumefaciens cfu, decreased the accumulation of phenolics, and improved transformation frequency. Our results show the hypersensitive response of excision stress for the expression of defense response-related genes and synthesis of metabolites in grain legume chickpea against pathogen infestation including Agrobacterium.

  12. CRESCIMENTO E OXIDAÇÃO DE EXPLANTES DE BANANEIRA-PRATA (Musa AAB IN VITRO: I. CONCENTRAÇÕES DE SAIS DE FERRO, COBRE E ZINCO

    Directory of Open Access Journals (Sweden)

    UTINO SERGIO

    2001-01-01

    Full Text Available Este experimento teve como objetivo avaliar a influência de diferentes concentrações de ferro, cobre e zinco do meio MS (MURASHIGE & SKOOG, 1962 no controle da oxidação de explantes iniciais de bananeira-Prata (Musa AAB. Foram utilizadas três concentrações (100, 50 e 0 miM de FeEDTA, duas concentrações (0,1 e 0miM de (CuSO4.5H2O e duas concentrações (30 e 0miM de (ZnSO4.7H2O, num delineamento inteiramente casualizado, arranjado em um fatorial completo 3 x 2 x 2, utilizando-se de 15 repetições. Ápices caulinares foram inoculados em meio MS modificado e, decorridos 28 dias após a inoculação, avaliaram-se a massa de matéria fresca, altura e grau de oxidação. Observou-se que esses micronutrientes são essenciais para o crescimento dos explantes e que a concentração de ferro influencia na oxidação de explantes, sendo que maiores graus de escurecimento foram observados nas concentrações mais elevadas. A redução ou retirada destes elementos do meio MS, isoladamente ou em combinações, não foi suficiente para eliminar a oxidação dos explantes.

  13. Post-explant visualization of thrombi in outflow grafts and their junction to a continuous-flow total artificial heart using a high-definition miniaturized camera.

    Science.gov (United States)

    Karimov, Jamshid H; Horvath, David; Sunagawa, Gengo; Byram, Nicole; Moazami, Nader; Golding, Leonard A R; Fukamachi, Kiyotaka

    2015-12-01

    Post-explant evaluation of the continuous-flow total artificial heart in preclinical studies can be extremely challenging because of the device's unique architecture. Determining the exact location of tissue regeneration, neointima formation, and thrombus is particularly important. In this report, we describe our first successful experience with visualizing the Cleveland Clinic continuous-flow total artificial heart using a custom-made high-definition miniature camera.

  14. Evaluación de diferentes combinaciones fitohormonales en la regeneración de Solanum tuberosum (Solanaceae Var. Pastusa Suprema a partir de explantes internodales

    Directory of Open Access Journals (Sweden)

    Jenny Paola Jiménez Barreto

    2009-07-01

    Full Text Available La regeneración de plantas mediante el cultivo de tejidos es un importante componente de la biotecnología que es requerido para procesos tales como la obtención de plantas transgénicas. Se estableció un sistema eficiente de regeneración para la especie Solanum tuberosum L. var. Pastusa Suprema, susceptible de ser transformada genéticamente. Se evaluó el efecto de las fitohormonas zeatina ribósido (ZR, ácido naftalénacetico (ANA y ácido gibérelico (AG3, utilizadas en combinaciones específicas, sobre la inducción de callo, la regeneración y el número de brotes producidos por explante. La presencia de ANA demostró ser esencial en la respuesta callogénica y regenerativa de los explantes. Se encontró que la adición de 3,0 mg/L de ZR, 0,02 mg/L de ANA y 1,0 mg/L de AG3 sobre un medio básico M-S, es una formulación hormonal adecuada para inducir el proceso de organogénesis indirecta sobre la variedad de papa Pastusa Suprema; produce callogénesis y regeneración en porcentajes superiores al 90%, con un promedio de seis regenerantes por explante.

  15. A morphometric and molecular study of the apoptosis observed on tadpoles' tail explants under the exposition of triiodothyronine in different homeopathic dilutions.

    Science.gov (United States)

    Guedes, José Roberto Pereira; Carrasco, Solange; Ferreira, Cláudia M; Bonamin, Leoni V; Goldenstein-Schainberg, Cláudia; Martins, Vanessa; Capelozzi, Vera L

    2016-08-01

    As a therapeutic system, homeopathy is supported by: i) similitude and experimentation in healthy individuals, ii) potentization. A challenge for researchers consists in looking for signals in water (or vehicle) to explain the storage of information in extremely high dilutions and the transfer of such information to the living systems. Anuran amphibian metamorphosis is controlled by thyroid hormones (TH), including the resorption of the tadpole tail. Apoptosis is a genetically regulated form of cell death that can be triggered by various extracellular and intracellular stimuli resulting in coordinated activation of a family of cysteine proteases called caspases. This study was blind and randomized. It performed in three stages: I) the identification of the most effective T3 homeopathic dilution to induce apoptotic reactions in Rana (Lithobates) catesbeianus tadpole tail explants stimulated by T3 in substantial, II) study of different controls and III) detection in explants under the action of the most effective dilution of T3, as established in Stage I. There was no statistically significant difference between tail macroscopic dimensions between the groups. T3 10cH decreased the expression of caspase 3/7 mRNA, in explants treated with T3 20 nM. The present experiment is in agreement with the hypothesis that T3, at a 10cH homeopathic dilution, changes the metamorphosis molecular network. Copyright © 2016 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.

  16. Suppression of MMP activity in bovine cartilage explants cultures has little if any effect on the release of aggrecanase-derived aggrecan fragments

    DEFF Research Database (Denmark)

    Wang, Bijue; Chen, Pingping; Jensen, Anne-Christine Bay

    2009-01-01

    BACKGROUND: Progressive loss of articular cartilage is a central hallmark in many joint disease, however, the relative importance of individual proteolytic pathways leading to cartilage erosion is at present unknown. We therefore investigated the time-dependant release ex vivo of MMP- and aggreca......BACKGROUND: Progressive loss of articular cartilage is a central hallmark in many joint disease, however, the relative importance of individual proteolytic pathways leading to cartilage erosion is at present unknown. We therefore investigated the time-dependant release ex vivo of MMP......- and aggrecanase-derived fragments of aggrecan and type II collagen into the supernatant of bovine cartilage explants cultures using neo-epitope specific immunoassays, and to associate the release of these fragments with the activity of proteolytic enzymes using inhibitors. FINDINGS: Bovine cartilage explants were...... cultured in the presence or absence of the catabolic cytokines oncostatin M (OSM) and tumor necrosis factor alpha (TNFalpha). In parallel, explants were co-cultured with protease inhibitors such as GM6001, TIMP1, TIMP2 and TIMP3. Fragments released into the supernatant were determined using a range of neo...

  17. HSV-1 interaction to 3-O-sulfated heparan sulfate in mouse-derived DRG explant and profiles of inflammatory markers during virus infection.

    Science.gov (United States)

    Sharthiya, Harsh; Seng, Chanmoly; Van Kuppevelt, T H; Tiwari, Vaibhav; Fornaro, Michele

    2017-06-01

    The molecular mechanism of herpes simplex virus (HSV) entry and the associated inflammatory response in the nervous system remain poorly understood. Using mouse-derived ex vivo dorsal root ganglia (DRG) explant model and single cell neurons (SCNs), in this study, we provided a visual evidence for the expression of heparan sulfate (HS) and 3-O-sulfated heparan sulfate (3-OS HS) followed by their interactions with HSV-1 glycoprotein B (gB) and glycoprotein D (gD) during cell entry. Upon heparanase treatment of DRG-derived SCN, a significant inhibition of HSV-1 entry was observed suggesting the involvement of HS role during viral entry. Finally, a cytokine array profile generated during HSV-1 infection in DRG explant indicated an enhanced expression of chemokines (LIX, TIMP-2, and M-CSF)-known regulators of HS. Taken together, these results highlight the significance of HS during HSV-1 entry in DRG explant. Further investigation is needed to understand which isoforms of 3-O-sulfotransferase (3-OST)-generated HS contributed during HSV-1 infection and associated cell damage.

  18. Geldanamycin induces production of heat shock protein 70 and partially attenuates ototoxicity caused by gentamicin in the organ of Corti explants

    Directory of Open Access Journals (Sweden)

    Haupt Heidemarie

    2009-09-01

    Full Text Available Abstract Background Heat shock protein 70 (HSP70 protects inner ear cells from damage and death induced by e.g. heat or toxins. Benzoquinone ansamycin antibiotic geldanamycin (GA was demonstrated to induce the expression of HSP70 in various animal cell types. The aim of our study was to investigate whether GA induces HSP70 in the organ of Corti (OC, which contains the auditory sensory cells, and whether GA can protect these cells from toxicity caused by a common aminoglycoside antibiotic gentamicin. Methods To address these questions, we used the OC explants isolated from p3-p5 rats. As a read-out, we used RT-PCR, ELISA and immunofluorescence. Results We found that GA at the concentration of 2 μM efficiently induced HSP70 expression on mRNA and protein level in the OC explants. Confocal microscopy revealed that HSP70 induced by GA is expressed by hair cells and interdental cells of spiral limbus. Preincubation of explants with 2 μM GA prior to adding gentamicin (500 μM significantly reduced the loss of outer but not inner hair cells, suggesting different mechanisms of otoprotection needed for these two cell types. Conclusion GA induced HSP70 in the auditory sensory cells and partially protected them from toxicity of gentamicin. Understanding the molecular mechanisms of GA otoprotection may provide insights for preventative therapy of the hearing loss caused by aminoglycoside antibiotics.

  19. Differential Gene Expression in Explanted Human Retinal Pigment Epithelial Cells 24-Hours Post-Exposure to 532 nm, 3.0 ns Pulsed Laser Light and 1064 nm, 170 ps Pulsed Laser Light 12-Hours Post-Exposure: Results Compendium

    National Research Council Canada - National Science Library

    Obringer, John

    2004-01-01

    .... We assessed the sublethal insult to human retinal pigment epithelial cells using a cadaver organ donor explant system for genes differentially expressed 12 and 24 hours post- exposure using gene...

  20. TGF-ß1 enhances the BMP-2-induced chondrogenesis of bovine synovial explants and arrests downstream differentiation at an early stage of hypertrophy.

    Science.gov (United States)

    Shintani, Nahoko; Siebenrock, Klaus A; Hunziker, Ernst B

    2013-01-01

    Synovial explants furnish an in-situ population of mesenchymal stem cells for the repair of articular cartilage. Although bone morphogenetic protein 2 (BMP-2) induces the chondrogenesis of bovine synovial explants, the cartilage formed is neither homogeneously distributed nor of an exclusively hyaline type. Furthermore, the downstream differentiation of chondrocytes proceeds to the stage of terminal hypertrophy, which is inextricably coupled with undesired matrix mineralization. With a view to optimizing BMP-2-induced chondrogenesis, the modulating influences of fibroblast growth factor 2 (FGF-2) and transforming growth factor beta 1 (TGF-ß1) were investigated. Explants of bovine calf metacarpal synovium were exposed to BMP-2 (200 ng/ml) for 4 (or 6) weeks. FGF-2 (10 ng/ml) or TGF-ß1 (10 ng/ml) was introduced at the onset of incubation and was present either during the first week of culturing alone or throughout its entire course. FGF-2 enhanced the BMP-2-induced increase in metachromatic staining for glycosaminoglycans (GAGs) only when it was present during the first week of culturing alone. TGF-ß1 enhanced not only the BMP-2-induced increase in metachromasia (to a greater degree than FGF-2), but also the biochemically-assayed accumulation of GAGs, when it was present throughout the entire culturing period; in addition, it arrested the downstream differentiation of cells at an early stage of hypertrophy. These findings were corroborated by an analysis of the gene- and protein-expression levels of key cartilaginous markers and by an estimation of individual cell volume. TGF-ß1 enhances the BMP-2-induced chondrogenesis of bovine synovial explants, improves the hyaline-like properties of the neocartilage, and arrests the downstream differentiation of cells at an early stage of hypertrophy. With the prospect of engineering a mature, truly articular type of cartilage in the context of clinical repair, our findings will be of importance in fine-tuning the

  1. Suppression of MMP activity in bovine cartilage explants cultures has little if any effect on the release of aggrecanase-derived aggrecan fragments

    Directory of Open Access Journals (Sweden)

    Sondergaard Bodil-Cecilie

    2009-12-01

    Full Text Available Abstract Background Progressive loss of articular cartilage is a central hallmark in many joint disease, however, the relative importance of individual proteolytic pathways leading to cartilage erosion is at present unknown. We therefore investigated the time-dependant release ex vivo of MMP- and aggrecanase-derived fragments of aggrecan and type II collagen into the supernatant of bovine cartilage explants cultures using neo-epitope specific immunoassays, and to associate the release of these fragments with the activity of proteolytic enzymes using inhibitors. Findings Bovine cartilage explants were cultured in the presence or absence of the catabolic cytokines oncostatin M (OSM and tumor necrosis factor alpha (TNFα. In parallel, explants were co-cultured with protease inhibitors such as GM6001, TIMP1, TIMP2 and TIMP3. Fragments released into the supernatant were determined using a range of neo-epitope specific immunoassays; (1 sandwich 342FFGVG-G2 ELISA, (2 competition NITEGE373ELISA (3 sandwich G1-NITEGE373 ELISA (4 competition 374ARGSV ELISA, and (5 sandwich 374ARGSV-G2 ELISA all detecting aggrecan fragments, and (6 sandwich CTX-II ELISA, detecting C-telopeptides of type II collagen. We found that (1 aggrecanase-derived aggrecan fragments are released in the early (day 2-7 and mid phase (day 9-14 into the supernatant from bovine explants cultures stimulated with catabolic cytokines, (2 the release of NITEGE373 neo-epitopes are delayed compared to the corresponding 374ARGSV fragments, (3 the MMP inhibitor GM6001 did not reduce the release of aggrecanase-derived fragment, but induced a further delay in the release of these fragments, and finally (4 the MMP-derived aggrecan and type II collagen fragments were released in the late phase (day 16-21 only. Conclusion Our data support the model, that aggrecanases and MMPs act independently in the processing of the aggrecan molecules, and furthermore that suppression of MMP-activity had little if

  2. Evidence for CB2 receptor involvement in LPS-induced reduction of cAMP intracellular levels in uterine explants from pregnant mice: pathophysiological implications.

    Science.gov (United States)

    Salazar, Ana Inés; Carozzo, Alejandro; Correa, Fernando; Davio, Carlos; Franchi, Ana María

    2017-07-01

    What is the role of the endocannabinoid system (eCS) on the lipopolysaccharide (LPS) effects on uterine explants from 7-day pregnant mice in a murine model of endotoxin-induced miscarriage? We found evidence for cannabinoid receptor type2 (CB2) involvement in LPS-induced increased prostaglandin-F2α (PGF2α) synthesis and diminished cyclic adenosine monophosphate (cAMP) intracellular content in uterine explants from early pregnant mice. Genital tract infections by Gram-negative bacteria are a common complication of human pregnancy that results in an increased risk of pregnancy loss. LPS, the main component of the Gram-negative bacterial wall, elicits a strong maternal inflammatory response that results in embryotoxicity and embryo resorption in a murine model endotoxin-induced early pregnancy loss. We have previously shown that the eCS mediates the embryotoxic effects of LPS, mainly via CB1 receptor activation. An in vitro study of mice uterine explants was performed to investigate the eCS in mediating the effects of LPS on PGF2α production and cAMP intracellular content. Eight to 12-week-old virgin female BALB/c or CD1 (wild-type [WT] or CB1-knockout [CB1-KO]) mice were paired with 8- to 12-week-old BALB/c or CD1 (WT or CB1-KO) males, respectively. On day 7 of pregnancy, BALB/c, CD1 WT or CD1 CB1-KO mice were euthanized, the uteri were excised, implantation sites were removed and the uterine tissues were separated from decidual and embryo tissues. Uterine explants were cultured and exposed for an appropriate amount of time to different pharmacological treatments. The tissues were then collected for cAMP assay and PGF2α content determination by radioimmunoassay. In vitro treatment of uteri explants from 7-day pregnant BALB/c or CD1 (WT or CB1-KO) mice with LPS induced an increased production of PGF2α (P Investigaciones Científicas y Técnicas (PIP 2012/0061). Dr Carlos Davio was funded by Agencia Nacional para la Promoción Científica y Tecnológica (PICT 2013

  3. TGF-ß1 enhances the BMP-2-induced chondrogenesis of bovine synovial explants and arrests downstream differentiation at an early stage of hypertrophy.

    Directory of Open Access Journals (Sweden)

    Nahoko Shintani

    Full Text Available Synovial explants furnish an in-situ population of mesenchymal stem cells for the repair of articular cartilage. Although bone morphogenetic protein 2 (BMP-2 induces the chondrogenesis of bovine synovial explants, the cartilage formed is neither homogeneously distributed nor of an exclusively hyaline type. Furthermore, the downstream differentiation of chondrocytes proceeds to the stage of terminal hypertrophy, which is inextricably coupled with undesired matrix mineralization. With a view to optimizing BMP-2-induced chondrogenesis, the modulating influences of fibroblast growth factor 2 (FGF-2 and transforming growth factor beta 1 (TGF-ß1 were investigated.Explants of bovine calf metacarpal synovium were exposed to BMP-2 (200 ng/ml for 4 (or 6 weeks. FGF-2 (10 ng/ml or TGF-ß1 (10 ng/ml was introduced at the onset of incubation and was present either during the first week of culturing alone or throughout its entire course. FGF-2 enhanced the BMP-2-induced increase in metachromatic staining for glycosaminoglycans (GAGs only when it was present during the first week of culturing alone. TGF-ß1 enhanced not only the BMP-2-induced increase in metachromasia (to a greater degree than FGF-2, but also the biochemically-assayed accumulation of GAGs, when it was present throughout the entire culturing period; in addition, it arrested the downstream differentiation of cells at an early stage of hypertrophy. These findings were corroborated by an analysis of the gene- and protein-expression levels of key cartilaginous markers and by an estimation of individual cell volume.TGF-ß1 enhances the BMP-2-induced chondrogenesis of bovine synovial explants, improves the hyaline-like properties of the neocartilage, and arrests the downstream differentiation of cells at an early stage of hypertrophy. With the prospect of engineering a mature, truly articular type of cartilage in the context of clinical repair, our findings will be of importance in fine-tuning the

  4. Dictionary Based Segmentation in Volumes

    DEFF Research Database (Denmark)

    Emerson, Monica Jane; Jespersen, Kristine Munk; Jørgensen, Peter Stanley

    2015-01-01

    We present a method for supervised volumetric segmentation based on a dictionary of small cubes composed of pairs of intensity and label cubes. Intensity cubes are small image volumes where each voxel contains an image intensity. Label cubes are volumes with voxelwise probabilities for a given...... label. The segmentation process is done by matching a cube from the volume, of the same size as the dictionary intensity cubes, to the most similar intensity dictionary cube, and from the associated label cube we get voxel-wise label probabilities. Probabilities from overlapping cubes are averaged...... and hereby we obtain a robust label probability encoding. The dictionary is computed from labeled volumetric image data based on weighted clustering. We experimentally demonstrate our method using two data sets from material science – a phantom data set of a solid oxide fuel cell simulation for detecting...

  5. Compliance with Segment Disclosure Initiatives

    DEFF Research Database (Denmark)

    Arya, Anil; Frimor, Hans; Mittendorf, Brian

    2013-01-01

    Regulatory oversight of capital markets has intensified in recent years, with a particular emphasis on expanding financial transparency. A notable instance is efforts by the Financial Accounting Standards Board that push firms to identify and report performance of individual business units...... (segments). This paper seeks to address short-run and long-run consequences of stringent enforcement of and uniform compliance with these segment disclosure standards. To do so, we develop a parsimonious model wherein a regulatory agency promulgates disclosure standards and either permits voluntary...... by increasing transparency and leveling the playing field. However, our analysis also demonstrates that in the long run, if firms are unable to use discretion in reporting to maintain their competitive edge, they may seek more destructive alternatives. Accounting for such concerns, in the long run, voluntary...

  6. Segmental osteotomies of the maxilla.

    Science.gov (United States)

    Rosen, H M

    1989-10-01

    Multiple segment Le Fort I osteotomies provide the maxillofacial surgeon with the capabilities to treat complex dentofacial deformities existing in all three planes of space. Sagittal, vertical, and transverse maxillomandibular discrepancies as well as three-dimensional abnormalities within the maxillary arch can be corrected simultaneously. Accordingly, optimal aesthetic enhancement of the facial skeleton and a functional, healthy occlusion can be realized. What may be perceived as elaborate treatment plans are in reality conservative in terms of osseous stability and treatment time required. The close cooperation of an orthodontist well-versed in segmental orthodontics and orthognathic surgery is critical to the success of such surgery. With close attention to surgical detail, the complication rate inherent in such surgery can be minimized and the treatment goals achieved in a timely and predictable fashion.

  7. Segmented fuel and moderator rod

    International Nuclear Information System (INIS)

    Doshi, P.K.

    1987-01-01

    This patent describes a continuous segmented fuel and moderator rod for use with a water cooled and moderated nuclear fuel assembly. The rod comprises: a lower fuel region containing a column of nuclear fuel; a moderator region, disposed axially above the fuel region. The moderator region has means for admitting and passing the water moderator therethrough for moderating an upper portion of the nuclear fuel assembly. The moderator region is separated from the fuel region by a water tight separator

  8. Segmentation of sows in farrowing pens

    DEFF Research Database (Denmark)

    Tu, Gang Jun; Karstoft, Henrik; Pedersen, Lene Juul

    2014-01-01

    The correct segmentation of a foreground object in video recordings is an important task for many surveillance systems. The development of an effective and practical algorithm to segment sows in grayscale video recordings captured under commercial production conditions is described...

  9. Segmentation in local hospital markets.

    Science.gov (United States)

    Dranove, D; White, W D; Wu, L

    1993-01-01

    This study examines evidence of market segmentation on the basis of patients' insurance status, demographic characteristics, and medical condition in selected local markets in California in the years 1983 and 1989. Substantial differences exist in the probability patients may be admitted to particular hospitals based on insurance coverage, particularly Medicaid, and race. Segmentation based on insurance and race is related to hospital characteristics, but not the characteristics of the hospital's community. Medicaid patients are more likely to go to hospitals with lower costs and fewer service offerings. Privately insured patients go to hospitals offering more services, although cost concerns are increasing. Hispanic patients also go to low-cost hospitals, ceteris paribus. Results indicate little evidence of segmentation based on medical condition in either 1983 or 1989, suggesting that "centers of excellence" have yet to play an important role in patient choice of hospital. The authors found that distance matters, and that patients prefer nearby hospitals, moreso for some medical conditions than others, in ways consistent with economic theories of consumer choice.

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

    Energy Technology Data Exchange (ETDEWEB)

    Kamada, Mizuna; Mitsui, Youji, E-mail: y-mitsui8310@hb.tp1.jp; Kumazaki, Tsutomu; Kawahara, Yuta; Matsuo, Taira; Takahashi, Tomoko, E-mail: t-takahashi@kph.bunri-u.ac.jp

    2014-10-24

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

  11. Establishment and Characterization of Primary Cultures from Iranian Oral Squamous Cell Carcinoma Patients by Enzymatic Method and Explant Culture

    Directory of Open Access Journals (Sweden)

    Meysam Ganjibakhsh

    2017-10-01

    Full Text Available Objectives: Oral Squamous Cell Carcinoma (OSCC is the most frequent oral cancer worldwide. It is known as the eighth most common cancer in men and as the fifth most common cancer in women. Cytogenetic and biochemical studies in recent decades have emphasized the necessity of providing an appropriate tool for such researches. Cancer cell culture is a useful tool for investigations on biochemical, genetic, molecular and immunological characteristics of different cancers, including oral cancer. Here, we explain the establishment process of five primary oral cancer cells derived from an Iranian population.Materials and Methods: The specimens were obtained from five oral cancer patients. Enzymatic, explant culture and magnetic-activated cell sorting (MACS methods were used for cell isolation. After quality control tests, characterization and authentication of primary oral cancer cells were performed by short tandem repeats (STR profiling, chromosome analysis, species identification, and monitoring the growth, morphology and the expression of CD326 and CD133 markers.Results: Five primary oral cancer cells were established from an Iranian population. The flow cytometry results showed that the isolated cells were positive for CD326 and CD133 markers. Furthermore, the cells were free from mycoplasma, bacterial and fungal contamination. No misidentified or cross-contaminated cells were detected by STR analysis.Conclusions: Human primary oral cancer cells provide an extremely useful platform for studying carcinogenesis pathways of oral cancer in Iranian population. They may be helpful in explaining the ethnic differences in cancer biology and the individuality in anticancer drug response in future studies.

  12. Roentgenological diagnoss of central segmental lung cancer

    International Nuclear Information System (INIS)

    Gurevich, L.A.; Fedchenko, G.G.

    1984-01-01

    Basing on an analysis of the results of clinicoroentgenological examination of 268 patments roentgenological semiotics of segmental lung cancer is presented. Some peculiarities of the X-ray picture of cancer of different segments of the lungs were revealed depending on tumor site and growth type. For the syndrome of segmental darkening the comprehensive X-ray methods where the chief method is tomography of the segmental bronchi are proposed

  13. Review of segmentation process in consumer markets

    OpenAIRE

    Veronika Jadczaková

    2013-01-01

    Although there has been a considerable debate on market segmentation over five decades, attention was merely devoted to single stages of the segmentation process. In doing so, stages as segmentation base selection or segments profiling have been heavily covered in the extant literature, whereas stages as implementation of the marketing strategy or market definition were of a comparably lower interest. Capitalizing on this shortcoming, this paper strives to close the gap and provide each step...

  14. Market Segmentation from a Behavioral Perspective

    Science.gov (United States)

    Wells, Victoria K.; Chang, Shing Wan; Oliveira-Castro, Jorge; Pallister, John

    2010-01-01

    A segmentation approach is presented using both traditional demographic segmentation bases (age, social class/occupation, and working status) and a segmentation by benefits sought. The benefits sought in this case are utilitarian and informational reinforcement, variables developed from the Behavioral Perspective Model (BPM). Using data from 1,847…

  15. Parallel fuzzy connected image segmentation on GPU

    OpenAIRE

    Zhuge, Ying; Cao, Yong; Udupa, Jayaram K.; Miller, Robert W.

    2011-01-01

    Purpose: Image segmentation techniques using fuzzy connectedness (FC) principles have shown their effectiveness in segmenting a variety of objects in several large applications. However, one challenge in these algorithms has been their excessive computational requirements when processing large image datasets. Nowadays, commodity graphics hardware provides a highly parallel computing environment. In this paper, the authors present a parallel fuzzy connected image segmentation algorithm impleme...

  16. LIFE-STYLE SEGMENTATION WITH TAILORED INTERVIEWING

    NARCIS (Netherlands)

    KAMAKURA, WA; WEDEL, M

    The authors present a tailored interviewing procedure for life-style segmentation. The procedure assumes that a life-style measurement instrument has been designed. A classification of a sample of consumers into life-style segments is obtained using a latent-class model. With these segments, the

  17. The Process of Marketing Segmentation Strategy Selection

    OpenAIRE

    Ionel Dumitru

    2007-01-01

    The process of marketing segmentation strategy selection represents the essence of strategical marketing. We present hereinafter the main forms of the marketing statategy segmentation: undifferentiated marketing, differentiated marketing, concentrated marketing and personalized marketing. In practice, the companies use a mix of these marketing segmentation methods in order to maximize the proffit and to satisfy the consumers’ needs.

  18. Automatic segmentation of vertebrae from radiographs

    DEFF Research Database (Denmark)

    Mysling, Peter; Petersen, Peter Kersten; Nielsen, Mads

    2011-01-01

    Segmentation of vertebral contours is an essential task in the design of automatic tools for vertebral fracture assessment. In this paper, we propose a novel segmentation technique which does not require operator interaction. The proposed technique solves the segmentation problem in a hierarchical...... is constrained by a conditional shape model, based on the variability of the coarse spine location estimates. The technique is evaluated on a data set of manually annotated lumbar radiographs. The results compare favorably to the previous work in automatic vertebra segmentation, in terms of both segmentation...

  19. Color image Segmentation using automatic thresholding techniques

    International Nuclear Information System (INIS)

    Harrabi, R.; Ben Braiek, E.

    2011-01-01

    In this paper, entropy and between-class variance based thresholding methods for color images segmentation are studied. The maximization of the between-class variance (MVI) and the entropy (ME) have been used as a criterion functions to determine an optimal threshold to segment images into nearly homogenous regions. Segmentation results from the two methods are validated and the segmentation sensitivity for the test data available is evaluated, and a comparative study between these methods in different color spaces is presented. The experimental results demonstrate the superiority of the MVI method for color image segmentation.

  20. MOVING WINDOW SEGMENTATION FRAMEWORK FOR POINT CLOUDS

    Directory of Open Access Journals (Sweden)

    G. Sithole

    2012-07-01

    Full Text Available As lidar point clouds become larger streamed processing becomes more attractive. This paper presents a framework for the streamed segmentation of point clouds with the intention of segmenting unstructured point clouds in real-time. The framework is composed of two main components. The first component segments points within a window shifting over the point cloud. The second component stitches the segments within the windows together. In this fashion a point cloud can be streamed through these two components in sequence, thus producing a segmentation. The algorithm has been tested on airborne lidar point cloud and some results of the performance of the framework are presented.

  1. Organogênese in vitro a partir de explante caulinar na regeneração de clones de Eucalyptus grandis W. Hill ex Maiden X E. urophylla S. T. Blake In vitro regeneration from stem explants of Eucalyptus grandis x E. urophylla clones through organogenesis

    Directory of Open Access Journals (Sweden)

    Elisa Cristina Soares de Carvalho Alves

    2004-10-01

    Full Text Available Com o objetivo de testar a regeneração in vitro por organogênese a partir de explante caulinar de três clones híbridos de Eucalyptus grandis x Eucalyptus urophylla, foram avaliados os efeitos dos reguladores de crescimento TDZ [1-fenil-3-(1,2,3-tiadiazol-5-iluréia], BAP (6-benzilaminopurina e ANA (ácido naftalenoacético. De modo geral, pôde-se observar resposta diferenciada dos clones quanto a intensidade, textura, coloração e grau de oxidação dos calos, em função dos tratamentos com os reguladores de crescimento. Os melhores resultados de calejamento foram dos tratamentos com a combinação dos reguladores de crescimento TDZ (0,5 mg L-1 e ANA (0,1 mg L-1, obtendo-se 100% de calejamento no explante caulinar. Houve a formação de estruturas nodulares compactas, principalmente na extremidade dos explantes caulinares, sendo essas regiões responsáveis pela regeneração de gemas adventícias. Em relação à regeneração, a melhor resposta foi obtida com 1,0 mg L-1 BAP.With the objective of testing in vitro regeneration through organogenesis from stem explants of three hybrid clones of Eucalyptus grandis x Eucalyptus urophylla, the effects of the growth regulators TDZ [1-phenil-3-(1, 2, 3-thiadiazol-5-ilurea], BAP (6-benzilaminopurine and NAA (Naphthalene acetic acid were evaluated. In general, a differentiated clone response in relation to intensity, texture, color and oxidation degree of calli was observed due to growth regulator treatments. The best results in callus formation were recorded for the treatments with the combination TDZ (0,5 mg L-1 and ANA (0,1 mg L-1, achieving 100% of callus formation from the stem explants. Compact node structures appeared mainly in the extremities of the stem explants, being those areas responsible for the regeneration of adventitious buds. The best regeneration response was obtained with 1,0 mg L-1 BAP.

  2. Unsupervised Performance Evaluation of Image Segmentation

    Directory of Open Access Journals (Sweden)

    Chabrier Sebastien

    2006-01-01

    Full Text Available We present in this paper a study of unsupervised evaluation criteria that enable the quantification of the quality of an image segmentation result. These evaluation criteria compute some statistics for each region or class in a segmentation result. Such an evaluation criterion can be useful for different applications: the comparison of segmentation results, the automatic choice of the best fitted parameters of a segmentation method for a given image, or the definition of new segmentation methods by optimization. We first present the state of art of unsupervised evaluation, and then, we compare six unsupervised evaluation criteria. For this comparative study, we use a database composed of 8400 synthetic gray-level images segmented in four different ways. Vinet's measure (correct classification rate is used as an objective criterion to compare the behavior of the different criteria. Finally, we present the experimental results on the segmentation evaluation of a few gray-level natural images.

  3. Efficient graph-cut tattoo segmentation

    Science.gov (United States)

    Kim, Joonsoo; Parra, Albert; Li, He; Delp, Edward J.

    2015-03-01

    Law enforcement is interested in exploiting tattoos as an information source to identify, track and prevent gang-related crimes. Many tattoo image retrieval systems have been described. In a retrieval system tattoo segmentation is an important step for retrieval accuracy since segmentation removes background information in a tattoo image. Existing segmentation methods do not extract the tattoo very well when the background includes textures and color similar to skin tones. In this paper we describe a tattoo segmentation approach by determining skin pixels in regions near the tattoo. In these regions graph-cut segmentation using a skin color model and a visual saliency map is used to find skin pixels. After segmentation we determine which set of skin pixels are connected with each other that form a closed contour including a tattoo. The regions surrounded by the closed contours are considered tattoo regions. Our method segments tattoos well when the background includes textures and color similar to skin.

  4. Metric Learning for Hyperspectral Image Segmentation

    Science.gov (United States)

    Bue, Brian D.; Thompson, David R.; Gilmore, Martha S.; Castano, Rebecca

    2011-01-01

    We present a metric learning approach to improve the performance of unsupervised hyperspectral image segmentation. Unsupervised spatial segmentation can assist both user visualization and automatic recognition of surface features. Analysts can use spatially-continuous segments to decrease noise levels and/or localize feature boundaries. However, existing segmentation methods use tasks-agnostic measures of similarity. Here we learn task-specific similarity measures from training data, improving segment fidelity to classes of interest. Multiclass Linear Discriminate Analysis produces a linear transform that optimally separates a labeled set of training classes. The defines a distance metric that generalized to a new scenes, enabling graph-based segmentation that emphasizes key spectral features. We describe tests based on data from the Compact Reconnaissance Imaging Spectrometer (CRISM) in which learned metrics improve segment homogeneity with respect to mineralogical classes.

  5. Interferon Induced Focal Segmental Glomerulosclerosis

    Directory of Open Access Journals (Sweden)

    Yusuf Kayar

    2016-01-01

    Full Text Available Behçet’s disease is an inflammatory disease of unknown etiology which involves recurring oral and genital aphthous ulcers and ocular lesions as well as articular, vascular, and nervous system involvement. Focal segmental glomerulosclerosis (FSGS is usually seen in viral infections, immune deficiency syndrome, sickle cell anemia, and hyperfiltration and secondary to interferon therapy. Here, we present a case of FSGS identified with kidney biopsy in a patient who had been diagnosed with Behçet’s disease and received interferon-alpha treatment for uveitis and presented with acute renal failure and nephrotic syndrome associated with interferon.

  6. A contrario line segment detection

    CERN Document Server

    von Gioi, Rafael Grompone

    2014-01-01

    The reliable detection of low-level image structures is an old and still challenging problem in computer vision. This?book leads a detailed tour through the LSD algorithm, a line segment detector designed to be fully automatic. Based on the a contrario framework, the algorithm works efficiently without the need of any parameter tuning. The design criteria are thoroughly explained and the algorithm's good and bad results are illustrated on real and synthetic images. The issues involved, as well as the strategies used, are common to many geometrical structure detection problems and some possible

  7. Did Globalization Lead to Segmentation?

    DEFF Research Database (Denmark)

    Di Vaio, Gianfranco; Enflo, Kerstin Sofia

    Economic historians have stressed that income convergence was a key feature of the 'OECD-club' and that globalization was among the accelerating forces of this process in the long-run. This view has however been challenged, since it suffers from an ad hoc selection of countries. In the paper......, a mixture model is applied to a sample of 64 countries to endogenously analyze the cross-country growth behavior over the period 1870-2003. Results show that growth patterns were segmented in two worldwide regimes, the first one being characterized by convergence, and the other one denoted by divergence...

  8. Comparison of hepatic MDCT, MRI, and DSA to explant pathology for the detection and treatment planning of hepatocellular carcinoma

    Directory of Open Access Journals (Sweden)

    Lauren M. Ladd

    2016-12-01

    Full Text Available Background/Aims The diagnosis and treatment plan for hepatocellular carcinoma (HCC can be made from radiologic imaging. However, lesion detection may vary depending on the imaging modality. This study aims to evaluate the sensitivities of hepatic multidetector computed tomography (MDCT, magnetic resonance imaging (MRI, and digital subtraction angiography (DSA in the detection of HCC and the consequent management impact on potential liver transplant patients. Methods One hundred and sixteen HCC lesions were analyzed in 41 patients who received an orthotopic liver transplant (OLT. All of the patients underwent pretransplantation hepatic DSA, MDCT, and/or MRI. The imaging results were independently reviewed retrospectively in a blinded fashion by two interventional and two abdominal radiologists. The liver explant pathology was used as the gold standard for assessing each imaging modality. Results The sensitivity for overall HCC detection was higher for cross-sectional imaging using MRI (51.5%, 95% confidence interval [CI]=36.2-58.4% and MDCT (49.8%, 95% CI=43.7-55.9% than for DSA (41.7%, 95% CI=36.2-47.3% (P=0.05. The difference in false-positive rate was not statistically significant between MRI (22%, MDCT (29%, and DSA (29% (P=0.67. The sensitivity was significantly higher for detecting right lobe lesions than left lobe lesions for all modalities (MRI: 56.1% vs. 43.1%, MDCT: 55.0% vs. 42.0%, and DSA: 46.9% vs. 33.9%; all P<0.01. The sensitivities of the three imaging modalities were also higher for lesions ≥2 cm vs. <2 cm (MRI: 73.4% vs. 32.7%, MDCT: 66.9% vs. 33.8%, and DSA: 62.2% vs. 24.1%; all P<0.01. The interobserver correlation was rated as very good to excellent. Conclusion The sensitivity for detecting HCC is higher for MRI and MDCT than for DSA, and so cross-sectional imaging modalities should be used to evaluate OLT candidacy.

  9. Pathogenesis of Focal Segmental Glomerulosclerosis

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    Beom Jin Lim

    2016-11-01

    Full Text Available Focal segmental glomerulosclerosis (FSGS is characterized by focal and segmental obliteration of glomerular capillary tufts with increased matrix. FSGS is classified as collapsing, tip, cellular, perihilar and not otherwise specified variants according to the location and character of the sclerotic lesion. Primary or idiopathic FSGS is considered to be related to podocyte injury, and the pathogenesis of podocyte injury has been actively investigated. Several circulating factors affecting podocyte permeability barrier have been proposed, but not proven to cause FSGS. FSGS may also be caused by genetic alterations. These genes are mainly those regulating slit diaphragm structure, actin cytoskeleton of podocytes, and foot process structure. The mode of inheritance and age of onset are different according to the gene involved. Recently, the role of parietal epithelial cells (PECs has been highlighted. Podocytes and PECs have common mesenchymal progenitors, therefore, PECs could be a source of podocyte repopulation after podocyte injury. Activated PECs migrate along adhesion to the glomerular tuft and may also contribute to the progression of sclerosis. Markers of activated PECs, including CD44, could be used to distinguish FSGS from minimal change disease. The pathogenesis of FSGS is very complex; however, understanding basic mechanisms of podocyte injury is important not only for basic research, but also for daily diagnostic pathology practice.

  10. Carprofen inhibits the release of matrix metalloproteinases 1, 3, and 13 in the secretome of an explant model of articular cartilage stimulated with interleukin 1β.

    Science.gov (United States)

    Williams, Adam; Smith, Julia R; Allaway, David; Harris, Pat; Liddell, Susan; Mobasheri, Ali

    2013-01-01

    Arthritic diseases are characterized by the degradation of collagenous and noncollagenous extracellular matrix (ECM) components in articular cartilage. The increased expression and activity of matrix metalloproteinases (MMPs) is partly responsible for cartilage degradation. This study used proteomics to identify inflammatory proteins and catabolic enzymes released in a serum-free explant model of articular cartilage stimulated with the pro-inflammatory cytokine interleukin 1β (IL-1β). Western blotting was used to quantify the release of selected proteins in the presence or absence of the cyclooxygenase-2 specific nonsteroidal pro-inflammatory drug carprofen. Cartilage explant cultures were established by using metacarpophalangeal joints from horses euthanized for purposes other than research. Samples were treated as follows: no treatment (control), IL-1β (10 ng/ml), carprofen (100 μg/ml), and carprofen (100 μg/ml) + IL-1β (10 ng/ml). Explants were incubated (37°C, 5% CO2) over twelve day time courses. High-throughput nano liquid chromatography/mass spectrometry/mass spectrometry uncovered candidate proteins for quantitative western blot analysis. Proteoglycan loss was assessed by using the dimethylmethylene blue (DMMB) assay, which measures the release of sulfated glycosaminoglycans (GAGs). Mass spectrometry identified MMP-1, -3, -13, and the ECM constituents thrombospondin-1 (TSP-1) and fibronectin-1 (FN1). IL-1β stimulation increased the release of all three MMPs. IL-1β also stimulated the fragmentation of FN1 and increased chondrocyte cell death (as assessed by β-actin release). Addition of carprofen significantly decreased MMP release and the appearance of a 60 kDa fragment of FN1 without causing any detectable cytotoxicity to chondrocytes. DMMB assays suggested that carprofen initially inhibited IL-1β-induced GAG release, but this effect was transient. Overall, during the two time courses, GAG release was 58.67% ± 10.91% (SD) for IL-1

  11. Micropropagaión de stevia rebaudiana bertoni, un endulzante natural a través de explantes con meristemos pre existentes

    OpenAIRE

    Suárez, Isidro; Quintero, Irma R.

    2014-01-01

    Título en ingles: Micropropagation of  Stevia rebaudiana Bertoni, a natural sweetener, through pre existing meristem explants Titulo corto: Micropropagación de Stevia rebaudiana Bertoni Resumen: Las hojas de Stevia rebaudiana son fuente de esteviosidos y rebaudiosidos, sustancias endulzantes con bajo contenido calórico. La propagación sexual y clonal de estevia es difícil debido a la calidad de la semilla y el tamaño reducido de la planta. Para evaluar la multiplicación, brotes establecidos i...

  12. Enrofloxacin and Toltrazuril Are Able to Reduce Toxoplasma gondii Growth in Human BeWo Trophoblastic Cells and Villous Explants from Human Third Trimester Pregnancy

    Directory of Open Access Journals (Sweden)

    Rafaela J. da Silva

    2017-07-01

    Full Text Available Classical treatment for congenital toxoplasmosis is based on combination of sulfadiazine and pyrimethamine plus folinic acid. Due to teratogenic effects and bone marrow suppression caused by pyrimethamine, the establishment of new therapeutic strategies is indispensable to minimize the side effects and improve the control of infection. Previous studies demonstrated that enrofloxacin and toltrazuril reduced the incidence of Neospora caninum and Toxoplasma gondii infection. The aim of the present study was to evaluate the efficacy of enrofloxacin and toltrazuril in the control of T. gondii infection in human trophoblast cells (BeWo line and in human villous explants from the third trimester. BeWo cells and villous were treated with several concentrations of enrofloxacin, toltrazuril, sulfadiazine, pyrimethamine, or combination of sulfadiazine+pyrimethamine, and the cellular or tissue viability was verified. Next, BeWo cells were infected by T. gondii (2F1 clone or the ME49 strain, whereas villous samples were only infected by the 2F1 clone. Then, infected cells and villous were treated with all antibiotics and the T. gondii intracellular proliferation as well as the cytokine production were analyzed. Finally, we evaluated the direct effect of enrofloxacin and toltrazuril in tachyzoites to verify possible changes in parasite structure. Enrofloxacin and toltrazuril did not decrease the viability of cells and villous in lower concentrations. Both drugs were able to significantly reduce the parasite intracellular proliferation in BeWo cells and villous explants when compared to untreated conditions. Regardless of the T. gondii strain, BeWo cells infected and treated with enrofloxacin or toltrazuril induced high levels of IL-6 and MIF. In villous explants, enrofloxacin induced high MIF production. Finally, the drugs increased the number of unviable parasites and triggered damage to tachyzoite structure. Taken together, it can be concluded that

  13. Enrofloxacin and Toltrazuril Are Able to Reduce Toxoplasma gondii Growth in Human BeWo Trophoblastic Cells and Villous Explants from Human Third Trimester Pregnancy.

    Science.gov (United States)

    da Silva, Rafaela J; Gomes, Angelica O; Franco, Priscila S; Pereira, Ariane S; Milian, Iliana C B; Ribeiro, Mayara; Fiorenzani, Paolo; Dos Santos, Maria C; Mineo, José R; da Silva, Neide M; Ferro, Eloisa A V; de Freitas Barbosa, Bellisa

    2017-01-01

    Classical treatment for congenital toxoplasmosis is based on combination of sulfadiazine and pyrimethamine plus folinic acid. Due to teratogenic effects and bone marrow suppression caused by pyrimethamine, the establishment of new therapeutic strategies is indispensable to minimize the side effects and improve the control of infection. Previous studies demonstrated that enrofloxacin and toltrazuril reduced the incidence of Neospora caninum and Toxoplasma gondii infection. The aim of the present study was to evaluate the efficacy of enrofloxacin and toltrazuril in the control of T. gondii infection in human trophoblast cells (BeWo line) and in human villous explants from the third trimester. BeWo cells and villous were treated with several concentrations of enrofloxacin, toltrazuril, sulfadiazine, pyrimethamine, or combination of sulfadiazine+pyrimethamine, and the cellular or tissue viability was verified. Next, BeWo cells were infected by T. gondii (2F1 clone or the ME49 strain), whereas villous samples were only infected by the 2F1 clone. Then, infected cells and villous were treated with all antibiotics and the T. gondii intracellular proliferation as well as the cytokine production were analyzed. Finally, we evaluated the direct effect of enrofloxacin and toltrazuril in tachyzoites to verify possible changes in parasite structure. Enrofloxacin and toltrazuril did not decrease the viability of cells and villous in lower concentrations. Both drugs were able to significantly reduce the parasite intracellular proliferation in BeWo cells and villous explants when compared to untreated conditions. Regardless of the T. gondii strain, BeWo cells infected and treated with enrofloxacin or toltrazuril induced high levels of IL-6 and MIF. In villous explants, enrofloxacin induced high MIF production. Finally, the drugs increased the number of unviable parasites and triggered damage to tachyzoite structure. Taken together, it can be concluded that enrofloxacin and

  14. Respiratory syncytial virus infection in sheep bronchial explants is associated with enhanced ETB receptor-mediate contractile functional and autoradiographic studies

    International Nuclear Information System (INIS)

    Fernandes, L.B.; D'Aprile, A.C.; Betts, R.J.; Goldie, R.G.

    2001-01-01

    Full text: Respiratory syncytial virus (RSV) is an important precipitant of asthma in children. The impact of RSV infection on endothelin (ET) receptor density and function in airways is unknown. In the present study, sheep bronchial rings were maintained as explants in culture for up to 48 h. During this time, both the structural integrity of the epithelium and carbachol responsiveness were preserved. Bronchial rings in culture were exposed to non-infected culture medium or to RSV (1/50 TCID 50 ) for 0, 24 and 48 h which caused marked damage to and loss of the epithelium. RSV infection did not significantly alter responsiveness to ET-1 at either 24 (Control EC 40 = 102 nM, 95% confidence limits, 76-138 nM vs RSV EC 40 = 66 nM, 95% confidence limits, 48-91 nM, n=5-6, P>0.05) or 48 h (Control EC 40 35 nM, 95% confidence limits, 19-66 nM vs RSV EC 40 = 55 nM, 95% confidence limits, 32-93 nM, n=8, P>0.05). As seen previously (Goldie et al., 1994), sarafotoxin S6c (StxS6c, ET B -selective) did not cause contraction in non-infected sheep bronchial explants. In contrast, StxS6c (300 nM) increased tone by 8±3% carbachol Emax (n=6-8) in explants exposed to RSV for 24 or 48 h. Light microscopic autoradiography was used to determine the relative distribution of ET A and ET B receptors using [ 125 I]-ET-1, BQ-123 (ET A -selective) and StxS6c. Sheep airway smooth muscle contains a homogeneous population of ET A receptors (Goldie et al., 1994). Since StxS6c caused significant contraction in RSV-infected bronchial explants, it was surprising that autoradiographic techniques failed to detect airway smooth muscle ET B receptors in these preparations. It is likely that ET B receptors fell below the level of detection of autoradiography. The significant StxS6c-induced contraction of sheep bronchi suggests the novel expression of ET B receptors triggered by RSV which might be relevant to RSV-associated asthma. Copyright (2001) Australasian Society of Clinical and Experimental

  15. Brain Tumor Image Segmentation in MRI Image

    Science.gov (United States)

    Peni Agustin Tjahyaningtijas, Hapsari

    2018-04-01

    Brain tumor segmentation plays an important role in medical image processing. Treatment of patients with brain tumors is highly dependent on early detection of these tumors. Early detection of brain tumors will improve the patient’s life chances. Diagnosis of brain tumors by experts usually use a manual segmentation that is difficult and time consuming because of the necessary automatic segmentation. Nowadays automatic segmentation is very populer and can be a solution to the problem of tumor brain segmentation with better performance. The purpose of this paper is to provide a review of MRI-based brain tumor segmentation methods. There are number of existing review papers, focusing on traditional methods for MRI-based brain tumor image segmentation. this paper, we focus on the recent trend of automatic segmentation in this field. First, an introduction to brain tumors and methods for brain tumor segmentation is given. Then, the state-of-the-art algorithms with a focus on recent trend of full automatic segmentaion are discussed. Finally, an assessment of the current state is presented and future developments to standardize MRI-based brain tumor segmentation methods into daily clinical routine are addressed.

  16. A new framework for interactive images segmentation

    International Nuclear Information System (INIS)

    Ashraf, M.; Sarim, M.; Shaikh, A.B.

    2017-01-01

    Image segmentation has become a widely studied research problem in image processing. There exist different graph based solutions for interactive image segmentation but the domain of image segmentation still needs persistent improvements. The segmentation quality of existing techniques generally depends on the manual input provided in beginning, therefore, these algorithms may not produce quality segmentation with initial seed labels provided by a novice user. In this work we investigated the use of cellular automata in image segmentation and proposed a new algorithm that follows a cellular automaton in label propagation. It incorporates both the pixel's local and global information in the segmentation process. We introduced the novel global constraints in automata evolution rules; hence proposed scheme of automata evolution is more effective than the automata based earlier evolution schemes. Global constraints are also effective in deceasing the sensitivity towards small changes made in manual input; therefore proposed approach is less dependent on label seed marks. It can produce the quality segmentation with modest user efforts. Segmentation results indicate that the proposed algorithm performs better than the earlier segmentation techniques. (author)

  17. Hierarchical image segmentation for learning object priors

    Energy Technology Data Exchange (ETDEWEB)

    Prasad, Lakshman [Los Alamos National Laboratory; Yang, Xingwei [TEMPLE UNIV.; Latecki, Longin J [TEMPLE UNIV.; Li, Nan [TEMPLE UNIV.

    2010-11-10

    The proposed segmentation approach naturally combines experience based and image based information. The experience based information is obtained by training a classifier for each object class. For a given test image, the result of each classifier is represented as a probability map. The final segmentation is obtained with a hierarchial image segmentation algorithm that considers both the probability maps and the image features such as color and edge strength. We also utilize image region hierarchy to obtain not only local but also semi-global features as input to the classifiers. Moreover, to get robust probability maps, we take into account the region context information by averaging the probability maps over different levels of the hierarchical segmentation algorithm. The obtained segmentation results are superior to the state-of-the-art supervised image segmentation algorithms.

  18. Image Segmentation Using Minimum Spanning Tree

    Science.gov (United States)

    Dewi, M. P.; Armiati, A.; Alvini, S.

    2018-04-01

    This research aim to segmented the digital image. The process of segmentation is to separate the object from the background. So the main object can be processed for the other purposes. Along with the development of technology in digital image processing application, the segmentation process becomes increasingly necessary. The segmented image which is the result of the segmentation process should accurate due to the next process need the interpretation of the information on the image. This article discussed the application of minimum spanning tree on graph in segmentation process of digital image. This method is able to separate an object from the background and the image will change to be the binary images. In this case, the object that being the focus is set in white, while the background is black or otherwise.

  19. Toxic Anterior Segment Syndrome (TASS

    Directory of Open Access Journals (Sweden)

    Özlem Öner

    2011-12-01

    Full Text Available Toxic anterior segment syndrome (TASS is a sterile intraocular inflammation caused by noninfectious substances, resulting in extensive toxic damage to the intraocular tissues. Possible etiologic factors of TASS include surgical trauma, bacterial endotoxin, intraocular solutions with inappropriate pH and osmolality, preservatives, denatured ophthalmic viscosurgical devices (OVD, inadequate sterilization, cleaning and rinsing of surgical devices, intraocular lenses, polishing and sterilizing compounds which are related to intraocular lenses. The characteristic signs and symptoms such as blurred vision, corneal edema, hypopyon and nonreactive pupil usually occur 24 hours after the cataract surgery. The differential diagnosis of TASS from infectious endophthalmitis is important. The main treatment for TASS formation is prevention. TASS is a cataract surgery complication that is more commonly seen nowadays. In this article, the possible underlying causes as well as treatment and prevention methods of TASS are summarized. (Turk J Oph thal mol 2011; 41: 407-13

  20. Short communication. N-(2-chloro-4-pyridyl)-N-phenylurea (4-CPPU) enhances in vitro direct shoot organogenesis of Citrus aurantium L. epicotyl segments compared to other commonly used cytokinins

    Energy Technology Data Exchange (ETDEWEB)

    Roussos, P. A.; Dimitriou, G.; Voloudakis, A. E.

    2011-07-01

    The effect of three concentrations of five different cytokinins, i.e. 6-benzylamino purine, 2-isopentyl adenine, kinetin (Kin), thidiazuron and N-(2-chloro-4-pyridyl)-N-phenylurea (4-CPPU), was evaluated on the in vitro direct shoot organogenesis of epicotyl explants of sour orange (Citrus aurantium L.). The basal medium used was that of Murashige and Tucker and epicotyl explants were incubated in medium supplemented with the prementioned cytokinins for 45 days. The addition of Kin and 4-CPPU in the medium enhanced the direct shoot organogenesis of sour orange epicotyl segments. The concentration of each of these two cytokinins which gave the best results, was combined with indole-3-acetic acid (IAA) or {alpha}-naphthalene acetic acid ({alpha}-NAA) at concentrations ranging from 0.01 mg L{sup -}1 to 0.2 mg L{sup -}1. The inclusion of IAA at 0.2 mg L{sup -}1 in the medium with 4-CPPU at 0.05 mg L{sup -}1 resulted in 100% successful direct shoot organogenesis, while the combination of Kin at 0.25 mg L{sup -}1 with IAA or {alpha}-NAA each at 0.01 mg L{sup -}1 presented equally high organogenesis percentages (91.7%). The incubation of the produced shoots, in medium supplemented with either indole-3-butyric acid or {alpha}-NAA resulted in high rooting percentages (up to 90%) and the rooted explants were successfully acclimatized under mist (85%). Although 4-CPPU has been used in in vitro culture of various species, this is the first report on its use in the direct shoot organogenesis of citrus species and could be of great value in citrus genetic transformation protocols using epicotyl segments, since this cytokinin resulted in the absolute organogenesis percentage. (Author) 20 refs.