• Journal of Plant Biotechnology
• /
• v.7 no.3
• /
• pp.161-167
• /
• 2005

Brassinosteroids are steroidal plant hormones and are known to modulate physiological and cellular response in a wide range of plant species. Considerable insights has been achieved of the physiological role of brassinosteroid in Brassica species in the past few years, but their effect on direct organogenesis has not been extensively studied. In this sense, under optimal basal media and growth conditions we tested the cellular and organogenic response of 24-epibrassinolide (EBL) in a variable concentration (0.1 to $5.0\;{\mu}M$) and Zeatin (Z) (1.0 to $100\;{\mu}M$) and their synergic effect on hypocotyl explants of cauliflower and broccoli. The isolated EBL accelerated cell elongation and promotes direct organogenesis. One micromolar EBL + $10\;{\mu}M$ of Z was the most efficient combination for cell elongation, cell differentiation as well as for organogenesis. A suppressing effect on root induction was confirmed for all the tested hormone levels. The general results indicate a synergic effect of EBL-Z and EBL potentates Zeatin activity, at least in certain tissues. Besides de genetic factors, we can speculate that the natural hormone concentration in the explants might affect the responses by application of exogenous growth regulators. Experiments with new plant growth regulators, like brassinolide, are important aiming to maximize or accelerate plant regeneration for in vitro multiplication or for genetic transformation.

• Journal of Periodontal and Implant Science
• /
• v.28 no.2
• /
• pp.291-310
• /
• 1998

This study was performed to evaluate the effects of extracts of Drynariae Rhizoma on the characteristics of rat calvaria cells(RCV) and bone marrow cells(RBM) which have the important role on the bone formation in vitro. Drynariae Rhizoma has been known as the useful herbal medicament for treatment of the wound healing including regeneration of bone fracture, and also has been used to treat the periodontal lesions, tooth mobility, gingival bleeding and pus discharge via sulcus in Oriental Medicine. In control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 10% fetal bovine serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, extracts of Drynariae Rhizoma(0.1, 1, 5, 10, $50{\mu}g/ml$) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 3, 7, 14, 21, 30th day, the amount of total protein synthesis and alkaline phosphatase activity of RCV at 2,4th day and those of RBM at 3, 6th day. And also, the calcified nodule of RCV was examed at 3, 5th day in three goup, control, experimental, culture with the PDGF group. The results were as follow ; 1. Both RCV and RBM cells in Drynariae Rhizoma-treated experimental group proliferated more rapidly than nontreated control group. The experimental group below $5{\mu}g/ml$ Drynariae Rhizoma-treated showed more prominent cell proliferation from the 7th day to the 21st day than the control group and above $10\;{\mu}g/ml$ treated group in RCV. 2. Amount of total protein synthesis was more increased in Drynariae Rhizomatreated group than in control group. In $5{\mu}g/ml$ Drynariae Rhizoma-treated group showed most prominent protein synthesis of the any other exrperimental group and control group. 3. Alkaline phosphatase activity also more increased in Drynariae Rhizomatreated group than control group. 4. Mineralized nodules in Drynariae Rhizoma-treated group were more than not in control group but also in PDGF-treated group. From the above results, Drynariae Rhizoma appeared to enhanced the proliferation, protein synthesis, alkaline phosphatase activity and cellular ability of mineralized nodule formation than PDGF. So that, we conclude that Drynariae Rhizoma enhances the activities of bone cells which have the important role on the periodontal regeneration and optimal application of Drynariae Rhizoma was thought to be useful as the means in bone regeneration.

• Journal of the Korea Organic Resources Recycling Association
• /
• v.23 no.1
• /
• pp.70-81
• /
• 2015

Helicases are known to be a proteins that use the chemical energy of NTP binding and hydrolyze to separate the complementary strands of double-stranded nucleic acids to single-stranded nucleic acids. They participate in various cellular metabolism in many organisms. DEAD-box proteins are ATP-dependent RNA helicase that participate in all biochemical steps involving RNA. DEAD-box3 (DDX3) gene is belonging to the DEAD-box family and plays an important role in germ cell development in many organisms including not only vertebrate, but also invertebrate during asexual and sexual reproduction and participates in stem cell differentiation during regeneration. In this study, in order to identify and characterize DDX3 gene in the earthworm, Perionyx excavatus having a powerful regeneration capacity, total RNA was isolated from adult head containing clitellum. Full length of DDX3 gene from P. excavatus, Pe-DDX3, was identified by RT-PCR using the total RNA from head as a template. Pe-DDX3 encoded a putative protein of 607 amino acids and it also has the nine conserved motifs of DEAD-box family, which is characteristic of DEAD-box protein family. It was confirmed that Pe-DDX3 has the nine conserved motifs by the comparison of entire amino acids sequence of Pe-DDX3 with other species of different taxa. Phylogenetic analysis revealed that Pe-DDX3 belongs to a DDX3 (PL10) subgroup of DEAD-box protein family. And it displayed a high homology with PL10a, b from P. dumerilii.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.3
• /
• pp.151-156
• /
• 1994

Acifluorfen-tolerant cell lines of S, ptycanthum were isolated by stepwise selection using suspension culture. Growth of unselected line was completely inhibited at $0.5\;\mu\textrm{M}$, while some selected lines grew at $8\;\mu\textrm{M}$ acifluorfen. After subculturing on acifluorfen-free medium for 4 passages, six of the eleven cell lines screened and maintained their tolerance to $2\;\mu\textrm{M}$ acifluorfen. The regeneration capacity of selected cell lines in Solanum ptycanthum differed depending on the tell line. The acifluorfen tolerance of the somarclones regenerated from acifluorfen-tolerant cell lines differed depending on the somarclone. When plants were heated with $16\;\mu\textrm{M}$ acifluorfen, unselected control plane had over 75% phytotoxicity Many selected cell lines had less phytotoxicity than the seed-grown control plants. Tolerance to acifluorfen was inherited to the self-pollinated progenies. The inheritance patterns differed depending on the clone. Acifluorfen tolerance was inherited as a semidominant trait. Other segregation patterns were also observed. acifluorfen tolerance was recessive and acifluorfen sensitivity was dominant.

• Journal of Periodontal and Implant Science
• /
• v.24 no.1
• /
• pp.51-63
• /
• 1994

There has been many attempts to develop a method that can regenerate periodontal tissues that were lost due to periodontal diseasd, but none of them was completely successful. This study was designed to investigate the healing and regeneration of periodontal tissue when bone substitutes such as porous replamineform hydroxyapatite and porous resorbable calcium carbonate were used in combination with oxidized cellulose membrane and collagen absorbable hemostat, compared to a control where only oxidized cellulose membrane or collagen absorbable hemostat were used. Chronic periodontitis was induced on mandibular premolars of and adult dog by placing orthodontic elastic ligatures for 10 weeks. After flap operation, the control group were received oxidized cellulose membrane (control- I )or collagen absorbable hemostat (control- II) only, while one experimental group was given either porous replamineform hydroxyapatite or porous resorbable calcium carbonate in addition to oxidized cellulose membrane (Experimental I-A, I-B), and another experimental group was treated by using either porous replamineform hydroxyapatite or porous resorbable calcium carbonate in addition to collagen absorbable hemostat. (Experimental II-A, II-B) After 56 weeks, healing was histologically analyzed with the following results. 1. Apical migration of junctional epithelium was observed only in areas coronal to the notch for both control and experimental group. 2. Inflammatory cell infiltration was not observed in any groups. 3. Oxidized cellulose membrane and collagen absorbable hemostat were completely resorbed. 4. Newly-formed cementum was observed up to the level where junctional epithelium was located, for both control and experimental groups. 5. Bone formation was limited of the middle portion of the notch in the control group, where as experimental groups showed bone formation up to the level of implant materials coronal to the notch. 6. Minute resorption of apically located portions of implanted materials was observed in experimental group I-B and II-B only.

• Animal cells and systems
• /
• v.16 no.3
• /
• pp.207-214
• /
• 2012

Burns are one of the most devastating forms of trauma and wound healing is a complex and multicellular process, which is executed and regulated by signaling networks involving numerous growth factors, cytokines, and chemokines. Recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) was specifically produced from rice cell culture through use of a recombinant technique in our laboratory. The effect of rhGM-CSF on promotion of deep second-degree burn wound healing on the back skin of a hamster model was evaluated through a randomized and double-blind trial. As macroscopic results, hamster skins of the experimental groups showed earlier recovery by new epidermis than the control groups. Immunohistochemical reactions of proliferating cell nuclear antigen and transforming growth factor-b1, which are indicators of cell proliferation, were more active in the experimental group, compared with the control group. On electron microscopy, basal cells in the epidermis of the experimental group showed oval nuclei, prominent nucleoli, numerous mitochondria and abundant free ribosomes. In addition, fibroblasts contained well-developed rough endoplasmic reticulum with dilated cisternae. Bundles of collagen fibrils filled the extracellular spaces. Particularly, ultrastructural features indicating active metabolism for regeneration of injured skin at 15 days after burn injury, including abundant euchromatin, plentiful free ribosomes, and numerous mitochondria, were observed. These findings suggest that use of rhGM-CSF could result in accelerated deep second-degree burn wound healing in animal models.

• Journal of Microbiology and Biotechnology
• /
• v.30 no.7
• /
• pp.996-1004
• /
• 2020

Various genetically engineered microorganisms have been developed for the removal of heavy metal contaminants. Metal biosorption by whole-cell biosorbents can be enhanced by overproduction of metal-binding proteins/peptides in the cytoplasm or on the cell surface. However, few studies have compared the biosorption capacity of whole cells expressing intracellular or surface-displayed metal-adsorbing proteins. In this study, several constructs were prepared for expressing intracellular and surface-displayed Ochrobactrum tritici 5bvl1 ChrB in Escherichia coli BL21(DE3) cells. E. coli cells expressing surface-displayed ChrB removed more Cr(VI) from aqueous solutions than cells with cytoplasmic ChrB under the same conditions. However, intracellular ChrB was less susceptible to variation in extracellular conditions (pH and ionic strength), and more effectively removed Cr(VI) from industrial wastewater than the surface-displayed ChrB at low pH (<3). An adsorption-desorption experiment demonstrated that compared with intracellular accumulation, cell-surface adsorption is reversible, which allows easy desorption of the adsorbed metal ions and regeneration of the bioadsorbent. In addition, an intrinsic ChrB protein fluorescence assay suggested that pH and salinity may influence the Cr(VI) adsorption capacity of ChrB-expressing E. coli cells by modulating the ChrB protein conformation. Although the characteristics of ChrB may not be universal for all metal-binding proteins, our study provides new insights into different engineering strategies for whole-cell biosorbents for removing heavy metals from industrial effluents.

• The Korean Journal of Mycology
• /
• v.21 no.4
• /
• pp.285-292
• /
• 1993

For the enzymatic conversion of 5'-XMP to 5'-GMP, partially purified XMP aminase from Escherichia coli was coupled with the yeast, Saccharomycrs cerevisiae, capable of ATP regeneration through glycolytic pathway. In order to elevate the level of XMP aminase in E. coli, $guaB^{-}(IMP\;dehydrogenase-less)$ mutant were introduced, and the yeast used as ATP supplier was treated by some method to increase its membrane permeability. The optimum conditions for efficient conversion reaction by energy-coupled system were investigated. As the results, a CH 41, $guaB^-$ mutant of E. coli K-12, showed 2.75 fold increase in the level of XMP aminase, compared with its parent cell. And the lyophylized yeast was the most effective at the ATP supplier. The optimum temperature and pH of conversion reaction were $40{\circ]C$ and pH 7.4, and the highest conversion ratio was shown under the reaction condition of 100 mM glucose, 100 mM inorganic phosphate and 6 mM AMP. When 36 units/ml XMP aminase used under the above conditions, the amount of 60 mg/ml yeast was sufficient to be used. Under the optimum condition, 71% of 1.8 mM(65.6 mg/100 ml) 5'-XMP was converted to 5'-GMP within 8 hr.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.3
• /
• pp.532-538
• /
• 2008

For the newly isolated $H_2$-producing chemoheterotrophic bacterium Citrobacter amalonaticus Y19, anaerobic glucose metabolism was studied in batch cultivation at varying initial glucose concentrations (3.5-9.5 g/l). The carbon-mass and energy balances were determined and utilized to analyze the carbon metabolic-pathways network. The analyses revealed (a) variable production of major metabolites ($H_2$, ethanol, acetate, lactate, $CO_2$, and cell mass) depending on initial glucose levels; (b) influence of NADH regeneration on the production of acetate, lactate, and ethanol; and (c) influence of the molar production of ATP on the production of biomass. The results reported in this paper suggest how the carbon metabolic pathway(s) should be designed for optimal Hz production, especially at high glucose concentrations, such as by blocking the carbon flux via lactate dehydrogenase from the pyruvate node.

• Applied Biological Chemistry
• /
• v.34 no.2
• /
• pp.134-141
• /
• 1991

In order to investigate the changes and characteristics of biochemical metabolic substances of soybean tissue culture during the cultural period, immature cotyledons were detached form the plant on 15th days after flowering and cultured in vitro for 3 weeks. The cultures were classified into embryogenic(EC) and non-embryogenic callus(NEC). A part of the EC lines were subcultured for another 3 weeks and classified into root forming(RFC), and shoot forming cultures(SFC). Another part of the EC lines were used for isolation of protoplasts, which were subsequently cultured in vitro for 4 weeks. The cultures were classified into embryogenic(PEC) and non-embryogenic callus(PNEC) derived from the protoplasts. The cultures of EC and PEC lines showed higher phenylalanine content and lower methionine content than those of NEC and PNEC. At organ differentiation stage, both cultures showed the content of aspartic acid decreased, while the other amino acids increased as a whole. The protein pattern analysis of the cultures revealed that EC and NEC lines contained distinctive polypeptides, with mass of ca. 18KD for EC and ca. 22KD for NEC respectively. The EC and PEC lines also showed high activity of peroxidase isozyme A(piA), while the RFC and SFC lines showed that of peroxidase isozyme B(piB).