In order to contribute to the Korean tea-plant culture and tea industry by means of increasing the production of tea-plants, I have performed the tissue culture of the organs of the anther, leaf and stem. As for the culture-material, I have used the anther of tea (Thea sinensis) at the tetrad uninucleate microspore stage and used medium of modified Murashige and Skoog as the basal medium supplemented with the growth regulators of NAA and 2, 4-D, yeast, kinetin and others at various concentrations. As for the handling of material, I have followed the common methods of sterilization and microtoming and paraffine imbedding method and observed systematically periodic changes of the microspores in culture. I have divided the leaf, stem and root into segments and sterilized them and used the modified Murashige and Skoog as the basal medium and observed the differentiation of roots and callus and the results are as follows. 1. In case of anther, I have found 2n callus was found in 30 out of 100 segments in M2 medium. 2. The differentiation of roots appeared in 24.5% of total leaf segments cultured and in 50.5% of stem and in 43.9% of root. 3. When the differentiation of stem in different parts was observed, the most frequent differentiation was found in the second part of all the 4 parts. 4. The most frequent formation of callus was noticed from the anther-walls in case of anther culture and from the veins in case of leaf culture. It is concluded that the seedlings of tea-plant could be multiplied most by means of tissue culture of the second part of the tea-plant stem and reduction in the expenditures of tea-plant propagation was possible through tissue culture.
Swelling properties and shear strength behavior of MgO-Sand mixtures with hydration procese of MgO are compared according to different MgO contents (WMgO/WTotal=0, 30, 50, 70, 100%) in this study. The specimens are prepared by mixing with crushed MgO refractory bricks and silica sand. After hydration, the particle size and the specific gravity of MgO were decreases. Through microstructure observation and X-ray diffraction analysis, it is confirmed that MgO changes from the cubic structure of Periclase to the hexagonal cubic structure of Brucite after hydration. As the MgO content increases, both swelling rate and swelling pressure of the mixtures increase. WMgO/WTotal=30% specimen shows relatively low swelling pressure and swelling rate because produced Mg(OH)2 mainly fills the pores between sand particles. However, in the case of MgO more than 50%, swelling pressure and swelling rate increase significantly because Mg(OH)2 fills the pores of sand particles at first and then either pushes out sand particles or Mg(OH)2 particles after filling the pores. As a result of the direct shear test, before hydration, the mixtures show a dilative behavior on high MgO contents and a contractive behavior on low MgO contents. However, after hydration, the behavior of all mixtures changes to contractive behavior. The threshold fraction of fine (i.e., Mg(OH)2) contents of the hydrated MgO-Sand mixtures reveals approximately 60% compared with normalized shear strength.
Background: Cytokine-mediated ex vivo expansion has been proposed as a means of increasing the number of cord blood (CB) hematopoietic stem cells for transplantation. As well as stem cell number, stromal cells are necessary for functional maturation of hematopoiesis. The purpose of this study was to analyze the development of stromal cells during ex vivo expansion of CB $CD34^+$ cells. Methods : $CD34^+$ cells were purified from CB by magnetic bead selection. The levels of of interleukin-3, interleukin-$1{\beta}$, interleukin-6, granulocyte macrophagecolony stimulating factor and tumor necrosis factor-${\alpha}$ were measured in culture supernatants on 0, 1, 2, and 3 weeks, using ELISA techniques. CB $CD34^+$ cells were expanded in Iscoves modified Dulbeccos medium in the presence of several cytokines. The expression of E-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, platelet/endothelial cell adhesion molecule-1, von Willebrand factor, vimentin, and CD14 in newly developed stromal cells was examined by immunocytochemical method. Relevant extracellular matrix (ECM) proteins and proper cytokines were also assayed for the most suitable condition for expansion of stromal cells. Results: Several cytokines were found to have been produced by CB $CD34^+$ cells as well as bone marrow-derived $CD34^+$ cells. During ex vivo expansion of CB $CD34^+$ cells, stromal cells appeared in the culture by day 4 and expanded over the following 7-10 days before being confluent by day 2 1. These cells expressed surface markers characteristic of cells of endothelial lineage. Furthermore, these stroaml cells also expanded effectively when treated with thrombopoietin+flt-3 ligand+stem cell factor+leukemia inhibitory factor or 0.1% poly-L-lysine-coated wells. Conclusion: Stromal cells were developed during ex vivo expansion of CB $CD34^+$ cells and that this development could be enhanced further by treating the stromal cells with cytokines or ECM.
In this study, we attempt to investigate the mechanisms by which PDL cells regulate osteoclast formation and also tc know whether PDL retained their characteristic phenotype during tooth eruption and interdental separation. Rats were prepared at developmental days 21 (pre-root formation), 27(toot development), 34(advanced root formation/eruption) and at later times(adult rats). To induce severe resorption state of alveolar bone and tooth root, interdental separation with brass wire was performed between the lower first and second molars for 2 weeks in adult rats. Rat mandibles were demineralized and embedded in paraffin, and horizontal and frontal section were prepared for immuno-histochemical analysis using PDL-specific protein 22 (PDLs22), receptor activator of NFKB ligand (RANKL) and osteoprotegerin (OPG) antibodies. 1. Root formation and eruption stage of tooth development. 1) PDLs22 immunolocalization was observed in tooth follicle/PDL cells and osteoblasts throught out the root formation and eruption stages of tooth development. 2) RANKL expression became stronger at eruption stage than root formation stage of tooth development. 3) Strong expression of OPG was detected in follice/PDL cells of toot formation stage but it was decreased with tooth eruption. 2. Interdental separation between lower first and second molar 1) Comparared to normal animal, multinucleated osteoclasts and odontoclasts were markedly induced in the alveolar bone and tooth root with PDL remodeling in hematoxylin-eosin section. 2) PDLs22 expression was decreased with interdental separation. 3) RANKL expression was Increased with interdental separation in PDL fibroblasts, osteoblasts, odontoclasts and it lacunae, resorting dentin, cementum and bone matrix. 4) OPG expression was slightly decreased in the PDL cells adjacent to the alveolar bone and root surface with interdental separation. These results suggested that during tooth eruption and tooth movement, RANKL and OPG in the periodontal tissues are important determinants regulating balanced alveolar bone and tooth root resorption. And it is also suggested that PDL cells retained their characteristic phenotype during tooth eruption and interdental separation except for the short period of PDL remodeling.
This study was designed to evaluate the expression of type I collagen in periodontal tissue during the experimental movement of rat incisors. Twenty-one Sprague-Dawley rats were divided into a control group(3 rats), and experimental groups(18 rats) where a force(75g) from helical springs across the maxillary incisors was applied. Experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. And tissue slides of control and experimental groups were studied histologically and immunohistochemically by LSAB(Labelled streptavidine Biotin) immunohistochemical staining for type I collagen. The results were as follows: 1. Until 28-day after force application, periodontal fibers were strectched on the tension side, and compressed in pressure side, and the arrangement of periodontal fibers was not recovered by that time. 2. The degree of type I collagen expression in control group was rare in the oral epithelium, predentin, pulp and periodontal ligament, but was mildly positive in osteoblasts, acellular cementum, cementoblasts, intermaxillary suture. 3. At acellular cementum of experimental group, the expression of type I collagen was moderate in 1-day and severe in 7-day, which was maintained until 28-day. 4. Type I collagen was observed in the newly formed fibrous connective tissue and osteoblasts at intermaxillary suture, moderately in 1-day, and severely in 14-day. 5. The tension side of periodontal ligament showed a more positive expression of type I collagen than the pressure side in 4-day. The degree was highest in 7-day and was not differentiated between sides in 14-day. 6. In the side wall of bone matrix on which osteoblasts were attached, type I collagen was expressed severely, especially in 7-day. From the above findings, we could suggest that bone remodeling in tooth movement be intimately related to the cell differentiation and the resulting formation of type I collagen.
Vitamin D is known to exert its action by activating DNA and RBA within target cells to produce proteins and enzymes that can be used in bone resorption process. Particularly, the active form of vitmain D, 1,25-dihydroxycholecalciferol $[1,25-(OH)_2D_3]$, is considered to be one of the most potent stimulators of osteoclatic acitivity in vitro. The purpose of this study was to evaluate the effect of 1,25-Dihydroxyvitamin $D_3$ on the avtivity of periodotal ligament cells and, the experimental tooth movement. Human periodontal ligament cells were collected from the first premolar tooth extracted for the orthodontic treatment, and were incubated in the environment of $37^{\circ}C$, 5% $CO_2$ and 95% humidity. Microtitration(MIT) assay was done at 10, 25, 50 and 100ng/ml of 1,25-Dihydroxyvitamin $D_3$. 21 Sprague-Daft rats were divided into a control gmup(3), and experimental groups(18) where 100g of force from helical spring was applied across the maxillary incisors 1,25-Dihydroxyvitamin $D_3$ was injected into periodontal ligament at the mesial or distal surface of maxillary incisors so that we can compare the control side and the experimental side. Expreimental groups were sac rifled at 12, 24, 36, 48, 72hours and 7 days after force application, respectively. And the obtained tissues were evaluated histologically. The observed results were as follows. 1. The activity of periodontal ligament cells in l0ng/ml or 25ng/ml of 1,25-Dihydroxyvitamin $D_3$ 1,25-Dihydroxyvitamin $D_3$ was not significantly different to the control at the cultivation of 1, 2 and 3 days. 2. The activity of periodontal ligament cells was significantly increased at 3 days in 50 ng/ml of 1,25-Dihydroxyvitamin $D_3$ and 2, 3 days in 100g/ml of 1,25-Dihydroxyvitamin $D_3$. 3. Up to 7 days after force application, there was no difference in osteoblastic activity, tearing of periodontal ligament and proliferation of capillary at tension side between 1,25-Dihydroxyvitamin $D_3$ injection side and the control side. 4. The osteoclastic activity and the resorption of alveolar bone was greater in 1,25-Dihydroxyvitamin $D_3$ injection side than the control side at 36 hours after force application.
This study was conducted to observe the early life history of sea bass, Lateolabrax japonicus. The fertilized eggs were spherical in shape and turned out to be separative and floated. Their membrane and yolk having 1~5 oil globule were transparent. Fertilized eggs were measured to be 1.33~1.46 mm in diameter. Hatching of eggs were started at 74 hrs 15 mins, 54 hrs 55 mins, 50 hrs 45 mins, after fertilization in water temperature $16.0^{\circ}C$, $18.0^{\circ}C$, $20.0^{\circ}C$ respectively. The newly hatching larvae were 3.79~3.97 mm in total length with 35~37 myomeres, and mouth and anus were closed. Melanophores were distributed on the up side of head, upper jaw and margins of the body. The 5 days after hatching larvae measured 4.78~5.24 mm in total length, yolk were completely absorbed, and transformed to postlarval stage. In this time, mouth of larvae was opened, and also melanophores were presented on the lower jaw. Head of larvae grew remarkably. The 21~22 days after hatching, total length of the larvae was 7.15~8.12 mm, the caudal fin rays began to differentiation. In 31 days after hatching, the larva were 8.46~9.16 mm in total length, and tip of the caudal notochord flexed $45^{\circ}$. The larvae reached to the juvenile stage with all the fins were developed at 61 days after hatching and attained 16.28~17.31 mm in total length.
By the investigation of "Sim-Kyoung-Bu-Ju" and the comparison study between the thought of "Sim-Kyoung-Bu-Ju" and that of Lee Je-ma, I've got the following conclusion. 1. All man have two mind. That is explained that "In-Sim" and "Do-Sim" in the "Sim-Kyong-Bu-Ju", "Kun-Ja-Ji-Sim" and "So-In-Ji-Sim" in the Lee Je-ma. 2. says that "In-Sim" and "Do-Sim" are the important point of the distinguishment of the "Sung-In" from "Jung-In". The "Sung-In" is the man who distinguishes "In-Sim" from "Do-Sim" well, and he always is cautious for "In-Sim"s" falling in desire by the "Do-Sim". In the case of LeeJe-ma,"Kun-Ja-Ji-Sim" is easy to know and "So-In-Ji-Sim" is hard to know. The man of "Kun-Ja-Ji-Sim" being large part is "Kun-Ja" and the man of "So-In-Ji-Sim" being large part is "So-In". 3. To reach the state of the "Kun-Ja", the "Sim-Kyoung-Bu-Ju" and Lee Je-ma present the variant training methods, "Kei-Shin-Kong-Ku" which they have in common. The "Sin-Kyoung-Bu-Ju" presents the "Kyung" firstly for "Kei-Shin-Kong-Ku", Lee Je-ma presents the "Yo-In-Sang-Jep-Ji-Sung" and "Ja-Ki-Tok-Tuk-Ji-Sung" for "Jel-Bu-Jel", "Jung-Bu-Jung", "Ji-In", "Ji-Chen". 4. The "Sim-Kyoung-Bu-Ju" says that establish the "Sung-Ui" by the "Kei-Shin-Kong-Ku", and to "Jung-Sim" by the "Sung-Ui", Lee Je-ma says "Chi-Sim-Jung-Ki" by "Ji-In".
The following results were obtained in this work intended for getting more informations on features of phosphorus in Korean characteristic virgin, upland and paddy soils for knowing the ways how to evaluate the soil phosphorus availability and utilize fertilizers efficiently. 1. A large part of soil phosphorus is occupied by inorganic phosphorus in the soils of which the cultivated soils showed higher accumulation of applied phosphorus as inorganic forms than the uncultivated soils. The accumulation of organic phosphorus was indicated the highest in the paddy soils, and the lowest in the uncultivated soils. Than the cultuivated soil, the uncultivated soils have higher C/P ratio which was estimated to be related to soil fertility status essentially. 2. Iron phosphate was shown to be the most dominant form in soil phosphorus. Difference in parent rocks, from which the soils were originated, was shown to affect the carious distribution of phosphorus forms in the uncultivated soils to a large extent. The paddy soils have almost same distribution pattern of soil phosphorus forms regardless of different parent rocks. 3. The different methods for available phosphorus estimation extracted different amounts of phosphorus from the soils. Close relations between the available phosphours extracted with the different methods and the amounts of seil inorganic phosphorus were shown : e.g. between Fe-P and available-P by Olsen method. 4. Phosphorus absorption coefficient correlated negatively to soil inorganic phosphorus, and also to available phosphorus extracted with the several methods.
Wang, Yiming;Wu, Jingni;Choi, Young Whan;Jun, Tae Hwan;Kwon, Soon Wook;Choi, In Soo;Kim, Yong Chul;Gupta, Ravi;Kim, Sun Tae
Journal of Life Science
/
v.25
no.10
/
pp.1091-1097
/
2015
We have isolated and characterized an ascorbate peroxidase (APx) gene, OsAPx1 from rice. Northern and Western blot analyses indicated that at young seedling stage, OsAPx1 mRNA was expressed highly in root, shoot apical meristem (SAM) and leaf sheath than leaf. In mature plant, OsAPx1 gene expressed highly in root, stem and flower but weakly in leaf. OsAPx1 gene and protein expression level was induced in leaves inoculated with Magnaporthe oryzae (M. oryzae) and Xanthomonas oryzae pv. oryzae (Xoo). Phytohormones treatment showed that OsAPx1 was up-regulated by jasmonic acid (JA), but was down regulated by ABA and SA co-treatments with JA, resulting that they have antagonistic effect on pathogen responsive OsAPx1 expression. Phylogenetic analysis illustrated that Arabidopsis AtAPx1 has a close relationship with OsAPx1. In AtAPx1 knock out lines, the accumulation of O2- and H2O2 are all highly detected than wild type, revealing that the high concentration of exogenous H2O2 cause the intercellular superoxide anion and hydrogen peroxide accumulation in AtAPx1 knockout plant. These results suggested that OsAPx1 gene may be associated with the pathogen defense cascades as the mediator for balancing redox state by acting ROS scavenger and is associated with response to the pathogen defense via Jasmonic acid signaling pathway.
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