Journal of The Korean Society of Grassland and Forage Science
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v.23
no.2
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pp.95-100
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2003
The alfalfa (Medicago sativa L.) callus was induced from seeds on SH medium contained $3\;mg/{\ell}$ of 2,4-D. Several regenerated alfalfa plants and many shoots were obtained by procedure of Kim et al. (1999); 1) incubation for 28~30 days on SH medium added $5\;mg/{\ell}$ of NAA and $2\;mg/{\ell}$ of Kinetin, 2) incubation for 3~5 days on SH medium added $11\;mg/{\ell}$ of 2,4-D and $1\;mg/{\ell}$ of kinetin. 3) incubation for 21~25 days on SH medium added $1.6\;g/{\ell}$ of ammonium sulfate and $5.75\;g/{\ell}$ of proline. To increase of root induction ratio on plant regeneration process of alfalfa, root induction ratio was examined on 8 kinds of medium, containing different amount of hormone and SH salt. Root induction ratio was higher on SH medium contained IBA than SH basal medium. In case 1.5 mg of IBA was added in SH medium, root induction ratio was the highest to 56.0% in this study. On the other hand, root induction ratio was higher on SH medium diminished SH salt amount to half volume and addition of IBA makes high root induction ratio, too. Thus, we conclude that the medium for root induction of alfalfa may be added $1.5\;mg/{\ell}$ of IBA and diminished SH salt amount to half volume.
Kim, Soung-Min;Lee, Jong-Ho;Jo, Joung-Ae;Lee, Seung-Cheol;Lee, Suk-Keun
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.31
no.5
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pp.440-453
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2005
Objectives : To develop a bioactive membrane for guided bone regeneration (GBR), the biocompatibility and bone regenerating capacity of the cellulose membrane obtained from the Ascidians squirt skin were evaluated. Materials and methods : After processing the pure cellulose membrane from the squirt skin, the morphological study, amino acid analysis and the immunoreactivity of the cellulose membrane were tested. Total eighteen male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into two control (n=8) and another two experimental groups (n=10). In the first experimental group (n=5), the cellulose membrane was applicated to the 8.0 mm sized calvarial bone defect and the same sized defect was left without cellulose membrane in the first control group (n=4). In the another experimental group (n=5), the cellulose membrane was applicated to the same sized calvarial bone defect after femoral bone graft and the same sized defect with bone graft was left without cellulose membrane in the another control group (n=4). Each group was sacrificed after 6 weeks, the histological study with H&E and Masson trichrome stain was done, and immunohistochemical stainings of angiogenin and VEGF were also carried out. Results : The squirt skin cellulose showed the bio-inductive effect on the bone and mesenchymal tissues in the periosteum of rat calvarial bone. This phenomenon was found only in the inner surface of the cellulose membrane after 6 weeks contrast to the outer surface. Bone defect covered with the bioactive cellulose membrane showed significantly greater bone formation compared with control groups. Mesenchymal cells beneath the inner surface of the bioactive cellulose membrane were positive to the angiogenin and VEGF antibodies. Conclusion : We suppose that there still remains extremely little amount of peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx. This composition could prevent the adverse immunological hypersensitivity and also induce bioactive properties of cellulose membrane. These properties induced the effective angiogenesis with rapid osteogenesis beneath the inner surface of cellulose membrane, and so the possibilities of clinical application in dental field as a GBR material will be able to be suggested.
Cacalia firma recently has been used increasingly as leaf vegetables but endangered in natural forest. In this work, we established the plant regeneration via adventitious shoot formation from petiole segments of seedling and in vitro plantlets. Wounding of seed coats and $GA_3$ treatments were effective to induce in vitro germination of seeds, whereas, seed did not germinate at all without these treatment. When cotyledon, leaf, petiole, and root segments of seedling were cultured on medium with 2 $mg{\cdot}L^{-1}$ benzyl adenine (BA) and 0.5 $mg{\cdot}L^{-1}$ naphthaleneacetic acid (NAA), petiole segments showed highest number of shoots per explant among the other segments. Among the various kinds of cytokinins, BA, isopentyl adenine (2-ip), kinetin, zeatin, thidiazuron (TDZ), TDZ and BA treatments were effective to induce high frequency of adventitious shoot formation from petiole segments of in vitro propagated plants. NAA stimulated the frequency of adventitious shoot formation but not for number of adventitious shoots per explants compared to TDZ or BA treatment alone. Most of adventitious shoots were developed directly from surfaces of explants. Adventitious shoots were transferred on medium with IBA for root formation, thereafter the plantlets were successfully transferred to soil.
The purpose of this study is to evaluate the phenomenon of attachment and spreading of the cultured rat calvarial cell inoculated on their surface of different kinds of biodegradable membrane which had been used on tissue regeneration on periodontal defects by using scanning electron microscope. In this experiment 30 Sprague-Dawley male rats (mean BW 150gm) were used to harvest abundant number of cell in the short period. The rats were sacrificed by decapitatioan to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Biodegradable barrier membrane were collected with collagen type, and were divided into 3 different kind of surface such as scattered, polarized and fine-net type as their surface texture. Microcover plate which usually used for cell culture was used as control for smooth surface. All the membrane were seeded with cultured calvarial cell on their surface. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. After the culture as designed time, all the membrane were washed with 0.1 M Phosphate Buffered saline and fuxed with 2.5% Glutaraldehyde. And all specimen were treated with $OsO_4$, and Tannic acid before drying the cell for coating the cell with gold. Scanning Electron Microscope was used to observation. The following results were obtained. I. During the whole period of experiment, the phenomenon of cell attachment and spreading were revealed similar pattern to compare with smooth surface culture plate and ordinary culture dish. 2. The shape of cell attachment and spreading on the surface of barrier membrane were observed no remarked difference pattern between smooth surface culture plate and ordinary culture dish. 3. The cytoplasmic process of cultured calvaria cell extent to the deep portion of barrier membrane like as their own proper shape. 4. There were no remarkable relationships between the degree of cultured cell spreading and surface structure of barrier membrane. 5. Slight starified layer of cultured calvaria cell were observed on the scattered type of resorbable membrane, Conclusively, this study thus suggest that cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of carrier for many cell which could be used as new tissue regeneration, and those tissue engeering technique may become an new method in the approach to the repair of bone defects.
To investigate optimal conditions for plant regeneration in Platycodon grandiflorum (Jacq. A. DC.).Both leaf and hypocotyl explants were cultured on Murashige& Skoog's (MS) medium supplemented with combinations of 0.1, 0.5, 1.0, or 2.0 mg/L cytokinins (BA and kinetin) and 1.0 mg/L 2,4-D for 6 weeks, respectively. According to the type of explant, the total shoot organogenesis (56.38%) in leaf explants was higher than in hypocotyls (28.20%). In comparison with kinetin and BA for the plant regeneration, the frequency (70.38%) of leaf explants was higher in combination with kinetin and 2,4-D than of BA with 2,4-D (42.38%), whereas the frequency (35.56%) of hypocotyls explants was higher in BA combination than kinetin combination (20.83%). Thehighest frequency (94.20%) was observed from the cultures of leaf explants on the MS medium supplemented with 1.0 mg/L kinetin and 1.0 mg/L 2,4-D. Upon transfer onto 1/2 MS basal medium containing 3% sucrose, shoots developed into plantlets with roots, and were well grown in soil in the greenhouse. These results lead us to speculate that the optimization of culture conditions was responsible for the mass propagation from in vitro cultures of Platycodon grandiflorum (Jacq. A. DC.).
The rh-BMP-4 is a subgroup of TGF-${\beta}$ superfamily. The application of rh-BMP in alveolar bony defect was reported to new alveolar bone and new cementum formation. For minimized complications following tooth replantation, a operator must replant a tooth fast at the pertinent position. This study was to evaluate the effect of rh-BMP-4 on periodontal regeneration and root resorption following tooth replantation in rats. The 50 Sprague-Dawley rats weighting about 130gm were used in this study. The animals were divided into three groups. Group 1 ; immediate replantation after extraction : Group 2 ; replantation stored teeth extraction of first molar, the removal of periodontal ligament with collagenase, and etching with citric acid : Group 3 ; replantation stored teeth with treated rh-BMP-4 in mesial root. Experimental animals were sacrificed 3, 7, 14 days after replantation by heart infusion. The maxillae were removed, fixed, demineralized, dehydrated, infiltrated and embedded with JB-4 mixture. For light microscopic observation, 5 micron sections were cut and stained with toluidine blue. The results of this study were as follows : 1. After experimental 3 days, all groups were observed dead space between periodontum and root. 2. After experimental 7 days, group 1 and group 3 were observed filling periodontal fibers between alveolar bone and root but group 2 were not. 3. After experimental 7 days, group 3 were observed appearance of attached cementoblast like cell on root surface. Group 1 were observed regular arrangement of fibroblasts and collagen fibers at ${\times}400$ observation. 4. After experimental 14 days, all group were observed filling periodontal fibers between alveolar bone and root. Group 1 were observed normal arrangement of periodontal fibers. Group 3 were observed less abnormal arrangement of periodontal fibers. Group 2 were not observed functional normal arrangement of periodontal fibers. 5. After experimental 14 days, group 2 and 3 were observed several root resorption and irregular root surface but group 1 were not. These results suggest that the rh-BMP-4 can stimulate cementogenesis and enhance to attach collagen fibers.
Kim, Ji Ah;Choi, I Song;Oh, Jong Min;Kim, Won Jae;Park, Jae Roh
Ecology and Resilient Infrastructure
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v.3
no.3
/
pp.201-206
/
2016
The purpose of this study is to find the regeneration method of filter media using physical and chemical treatment for restoration of phosphorus adsorption ability. The filtration material used in this study is called Adphos. In an experiment of heating treatment, re-used filter media is heated to a high temperature before the adsorption test. The results show that the $PO{_4}^{3-}-P$ adsorption capacity is in the range of 0.0021 - 0.0030 mg/g and the removal efficiency is in the range of 26.1 - 39.4%. In the experiment of acid or basic treatment, re-used filter media is exposed to a different pH condition before the adsorption test. The results show that the $PO_4^{3-}-P$ adsorption capacity is in the range of 0.0010 - 0.0066 mg/g and the removal efficiency is in the range of 15.8 - 87.1% after the acid treatments which have pH values of 1 - 5. However, after the basic treatments which have pH values of 8 - 11, the results show that the $PO{_4}^{3-}-P$ adsorption capacity is in the range of 0.0018 - 0.0034 mg/g and the removal efficiency is in the range of 26.7 - 48.0%. In an experiment of chemical treatment using NaCl, re-used filter media was exposed to a different NaCl concentration before the adsorption test. The results show that the $PO{_4}^{3-}-P$ adsorption capacity is in the range of 0.0036 - 0.0050 mg/g and the removal efficiency is in the range of 50.5 - 71.1%. In conclusion, chemical treatment using NaCl shows a high recovery probability of phosphorus adsorption ability of filter media.
Cho Mi-Ae;Park Yun-Ok;Kim Jin-Suck;Park Ki-Jin;Min Hwang-Ki;Liu Jang-Ryol;Choi Pil-Son
Journal of Plant Biotechnology
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v.32
no.2
/
pp.117-121
/
2005
Immature embryos of 3 cultivars (Du Me Chal, Mi Baek Chal, Heug Jeom Chal) and 5 genotypes (HW1, KL103, HW3, HW4, KW7) were cultured on medium containing MS salts, Eriksson's vitamins, 1 mg/L 2,4-dichlorophenoxyacetic acid, 25 mM proline, 100 mg/L casamino acid, 3 mM MES, 1.7 mg/L $AgNO_3$ and 20 g/L sucrose (SIM). Frequency of somatic embryo formation on explant of immature embryos showed in HW1 (45.20%), KL103 (5.75%), HW3 (37.20%), HW4 (30.10%), KW70 (55.20%), Mi Baek Chal (18.74%), Heug Jeom Chal (22.41%), Du Me Chal (36.72%) and Hi II type (<10%), respectively. Yellowish friable embryogenic callus (YFEC) such as type II callus of Hi II genotype only produced from the HW3 and Heug Jeom Chal, whereas other cultivars and genotypes were directly formed somatic embryos with late-embryonic stages or expanded yellowish compact somatic embryo with morphological abnormality. The yellowish friable embryogenic callus (YFEC) could be proliferated on the same medium, which were maintained embryogenic capacity for 6 months over. Upon transfer to first regeneration and second regeneration medium, somatic embryos converted to plantlets at a frequency of approximately 100%. However, the expanded somatic embryos with abnormal morphology were slowly proliferated when subcultured on the same medium, and some of them were degenerated or converted to plantlets at a frequency of approximately 25%. Accordingly, The Heug Jeom Chal and HW3 genotype will be further used for development of high frequency transformation system in domestic maize germplasm.
An experimental study was done on rabbits to observe the effects of several anticlonorchial drugs on the pathology of the liver infested with Clonorchis sinensis. After two months of infestation with Clonorchis sinensis by giving $400{\sim}500$ metacercariae by mouth, hexachlorophene, chloroquine 2,2' methylenebis (3,4,6 trichlorophenoxy acetic acid) and Hetol were administered orally and follow up macro-and microscopic studies of the liver pathology were done in 2 to 3 days, one month, 2 months and 3 months after the completion of medications. The results obtained in this study are as follows: 1. In both groups which were administered hexachlorophene piperazine 20mg/kg for seven days or 8mg/kg for 18 days, the macroscopic findings of the liver after 3 months revealed only minimal changes of the color and consistency The histopathological findings were the reduction of fibrosis, pseudolobulation, proliferation and adenomatous hyperplasia of bile ducts, and regeneration of liver parenchyma. 2. In groups which were administered chloroquine phosphate 20mg/kg for 18 days or 40mg/kg for 5 days, and also in groups which were administered dithiazanine iodide 30mg/kg for 18 days or 60mg/kg for 5 days, no significant findings of recovery were observed either macroscopically or microscopically. 3. In the group which was given 20mg/kg of 2,2' methylenebis(3,4,6 trichlorophenoxy acetic acid) for 5 days, prominent healing of the damaged tissues was observed after 2 months, revealing the decrease of fibrous tissue, caliber of bile ducts and adenomatous hyperplasia of the epitherial cells of the bile ducts, and regenerationof liver parenchyma. 4. In the group which was given Hetol 200mg/kg for 5 days, swelling, congestion and eddish-brown discoloration of the liver were noted macroscopically after 3 days of completion of drug administration. Hemorrhage, congestion, necrosis and degeneration of the parenchyma were observed microscopically After 10 days, liver appeared almost normal macroscopically, but marked fat degeneration was noted microscopically. After 2 months, the liver was almost normal in gross appearence with only slight atrophy and also marked healing was observed microscopically, i. e. decrease of fibrous tissue and reduction of the previously enlarged bile duct and the regeneration of the liver parenchyma.
Kim, Chi Moon;Kwon, Ki Won;Moon, Heung Kyu;Lee, Jae Soon
Korean Journal of Agricultural Science
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v.14
no.2
/
pp.205-212
/
1987
To examine the morphogenetic response, stem segments of 8 Populus spp. and 3 different explants of P. nigra var. italica were cultured on MS (Murashige and Skoog 1962) and WPM (Woody Plant Medium) medium containing various phytohormones. The results obtained were as follows: 1. Shoot regeneration and development from stem segment of 8 Populus spp. showed a quite difference according to the section and the species. All of the species of Leuce and Tacamahaca section did not form adventitious buds, while most of explants showed axillary or dormant bud elongation after 4 weeks. But P. nigra var. italica of Aigeiros section showed a successful adventitious bud formation (mean 5.4 buds per explant). 2. Leaf, petiole, and internode segment of P. nigra var. italica showed a quite differences according to media and ex plants upon the morphogenetic response. Adventitious bud formation from leaf was more abundant and readily initiated on the abaxial side than on the adaxial side. Mean number of 103 adventitious buds per explant was obtained from abaxial side of leaf segment cultured on WPM medium containing $0.2mg/{\ell}$ BAP for 5 weeks. 3. 2,4-D (2,4-dichlorophenoxy acetic acid) supplemented to media appeared to be negative upon the adventitious bud formation of P. nigra var. italica, while it promoted callus formation from all explants. Especially, NAA (${\alpha}$-naphtalene acetic acid) or NAA combination with BAP (6-benzylaminopurine) promoted root regeneration from the all explant of P. nigra var. italica in this study.
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