• Title/Summary/Keyword: Nodule formation

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Zinc Deficiency Decreased Alkaline Phosphatase Expression and Bone Matrix Ca Deposits in Osteoblast-like MC3T3-E1 Cells

  • Cho Young-Eon;Lomeda Ria-Ann R.;Kim Yang-Ha;Ryu Sang-Hoon;Choi Je-Yong;Kim Hyo-Jin;Beattie John H.;Kwun In-Sook
    • Nutritional Sciences
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    • v.8 no.4
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    • pp.242-249
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    • 2005
  • It is well established that zinc plays an important role in bone metabolism and mineralization. The role of zinc in bone formation is well documented in animal models, but not much reported in cell models. In the present study, we evaluated zinc deficiency effects on osteoblastic cell proliferation, alkaline phosphatase activity and expression, and extracellular matrix bone nodule formation and bone-related gene expression in osteoblastic MC3T3-E1 cells. To deplete cellular zinc, chelexed-FBS and interpermeable zinc chelator TPEN were used. MC3T3-E1 cells were cultured in zinc concentration-dependent (0-15 ${\mu}M\;ZnCl_2$) and time-dependent (0-20 days) manners. MC3T3-E1 cell proliferation by MTT assay was increased as medium zinc level increased (p<0.05). Cellular Ca level and alkaline phosphatase activity were increased as medium zinc level increased (p<0.05). Alkaline phosphatase expression, a marker of commitment to the osteoblast lineage, measured by alkaline phosphatase staining was increased as medium zinc level increased. Extracellular calcium deposits measured by von Kossa staining for nodule formation also appeared higher in Zn+(15 ${\mu}M\;ZnCl_2$) than in Zn-(0 ${\mu}M\;ZnCl_2$). Bone formation marker genes, alkaline phosphatase and osteocalcin, were also expressed higher in Zn+ than in Zn-. The current work supports the beneficial effect of zinc on bone mineralization and bone-related gene expression. The results also promote further study as to the molecular mechanism of zinc deficiency for bone formation and thus facilitate to design preventive strategies for zinc-deficient bone diseases.

Pulmonary Vessel Extraction and Nodule Reclassification Method Using Chest CT Images (흉부 CT 영상을 이용한 폐 혈관 추출 및 폐 결절 재분류 기법)

  • Kim, Hyun-Soo;Peng, Shao-Hu;Muzzammil, Khairul;Kim, Deok-Hwan
    • Journal of the Institute of Electronics Engineers of Korea CI
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    • v.46 no.6
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    • pp.35-43
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    • 2009
  • In the Computer Aided Diagnosis(CAD) System, the efficient way of classifying nodules from chest CT images of a patient is to perform the classification of the remaining part after the pulmonary vessel extraction. During the pulmonary vessel extraction, due to the small difference between the vessel and nodule features in imaging studies such as CT scans after having an injection of contrast, nodule maybe extracted along with the pulmonary vessel. Therefore, the pulmonary vessel extraction method plays an important role in the nodule classification process. In this paper, we propose a nodule reclassification method based on vessel thickness analysis. The proposed method consist of four steps, lung region searching step, vessel extraction and thinning step, vessel topology formation and correction step and the reclassification of nodule in the vessel candidate step. The radiologists helped us to compare the accuracy of the CAD system using the proposed method and the accuracy of general one. Experimental results show that the proposed method can extract pulmonary vessels and reclassify false-positive nodules accurately.

Effects of Chitosan on Human Periodontal Ligament Cells in Vitro (키토산이 배양중인 치주인대세포에 미치는 영향)

  • Kim, Ok-Su;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.163-180
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    • 2001
  • The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of periodontal ligament cells. Primary human periodontal ligament cells were cultured in dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells of 4th to 7th passage were inoculated in the multiwell plates coated with chitosan in concentration of 0.22, 0.2, and $2mg/m{\ell}$. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized modules was evaluated after 21 days of culture. The results were as follows : 1. The morphology of periodontal ligament cells on the chitosan coating was round or spheric. Round cells were aggregated after 6 hours of culture. Aggregated cells on the chitosan coated surface showed nodule-like appearance after 24 hours of culture and not achieved confluency at 7 days. 2. During early period of culture, the attachment of periodontal ligament cells were inhibited by chitosan coating. Inhibition of cell attachment tended to increase with the concentration of chitosan. 3. At the chitosan concentration of 0.02 and $0.2mg/m{\ell}$, periodontal ligament cells were more rapidly proliferated at 7 days, compared to the control group. At the concentration of $2mg/m{\ell}$, the proliferation of periodontal ligament cells was inhibitied(p<0.01). 4. Alkaline phosphatase activity of periodontal ligament cells was increased in chitosan coated group, especially at the concentration of $0.02mg/m{\ell}$after 4 days of culture.5. Periodontal ligament cells produced mineralized nodules on chitosan coated wells without the addition of mineralized nodule forming materials (ascorbic acid, ${\beta}-glycerophosphat$, dexamethasone). With the addition of mineralized nodule forming materials, periodontal ligament cells produced more mineralized nodules at the concentration of $0.02mg/m{\ell}$, compared to the control. In summary, the attachment, proliferation, cell activity, and alkaline phosphatase activity of periodontal ligament cells depended on the concentration of coated chitosan. Chitosan stimulated mineralized nodule formation by periodontal ligament cells. At the appropriate concentration($0.02mg/m{\ell}$), chitosan could increase alkaline phosphatase activity and stimulate the formation of mineralized nodule by periodontal ligament cells. These results suggest that chitosan can be used as an adjunct for bone graft material, and the matrix of tissue engineering for periodontal regeneration, especially bone regeneration.

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Physiological and biochemical studies on legunme nodule bacteria, Rhizobia -I. Some charateristics of isolated strains of Rhizobia and inoculation test on soy bean. (두과작물 근류균에 대한 생리 및 생화학적 연구 -I. 근류균 균주의 특성과 접종시험-)

  • Lim, Sun-Uk
    • Applied Biological Chemistry
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    • v.13 no.1
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    • pp.51-57
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    • 1970
  • On the basis of the specific interrelationship between the species or variety of leguminous crops and the species or strain of nodule bacteria, Rhizobia, the rhizobial species and strain must be effectively chosen for the successful inoculation. The present paper describes on some results of the isolation and taxonomic study on the native rhizobial strains isolated from the nodules of five species of leguminous crops such as numerous varieties of soy bean, lespedeza, birdfoot trefoil, ladino and red clovers. The isolated strains of soy bean nodule bacterium, Rhizobium japonicum were grouped through the inoculation test on variety Changdanbaikmock into the effective, noneffective and toxic strain for the nodule formation. In the study of the effect of some inorganic and organic nitrogenous compounds on the growth of Rhizobium japonicum strain Ac 20, a promotive response was showed by asparagine, and glutamine, but hydroxylamine, nitrite, hydrazine and azide was inhibitory at the concentration of $10^{-2}M/l$ in mannitol-yeast extract basal medium. In the physiological characteristics each strain showed somewhat different activities of the indole-3-actic acid formation and hydrogenase and discussed with these characters in relation to nodule forming ability.

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THE EFFECTS OF GINGIVAL FIBROBLAST ON THE MINERALIZATION OF THE RAT BONE MARROW STROMAL CELL (백서 골수세포의 석회화 과정에 미치는 치은 섬유아세포의 영향)

  • Kim, Seuk-Yong;Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.25 no.2
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    • pp.210-221
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    • 1995
  • The purpose of this study was performed to investigate the mineralization and differentiation of osteobalsts for bone regeneration in vitro and the effect of rate of the composition in periodontal cells on mineralization. For this study, healthy gingival tissues were surgically obtained from the patients during 1st premolar extraction for the purposes of orthodontic treament. Gingival tissue was washed several time with Phosphate buffered saline contained high concentration of antibiotics and antifungal agent, and cultured in Dulbecco's Modified Eagle's Medium(DMEM, Gibco, U.S.A.). Every cell were cultured in state at $37^{\circ}C$, 100% of humidity, 5% of $CO_2$ incubator. Bone marrow stromal cells were isolated from 5-clay-old rat femur with using medium irrigation mathod by syringe. Cell suspension medium were centrifuged at 1500 rpm for 5 min and then cultured in the petri dish. Two kinds of cell were freezed and stocked in the liquid nitrogen tank until experiment. Cell were incubated into the 24 multi-well plate with $5{\times}10^4$cell/well of medium at $37^{\circ}C$, 100% of humidity 5% $CO_2$ incubator for 24 hours. After discarded of the supernatent of medium, O.5ml of medium were reapplied and incubated. And counted the number of cell using the hemocytometer and inverted light microscope. We have measured the number of mineralized nodule with using Alizarin red S. staining in microscope. Furthermore every cell were observed the morphological change between every rate of co-culture of the two kinds of cell. The results were as follows; The rate of proliferation of co-culture cell revealed high rate tendency compared the bone marrow stromal cell only and low growth rate to compared with gingival fibroblast only. The tendency of formation of the mineralized nodule were observed dose-depend pattern of bone marrow stromal cell. It is concluded that the gingival fibroblast may inhibit the formation of mineralized nodule in the culture of the bone marrow stromal cell.

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Effects of Nitrogen and Potassium on Plant Growth, Yield and Nodule Formation in Winged Bean(Psophocarpus tetragonolobus(L.) D. C.) (질소와 가리시용이 Winged bean의 생육, 수량 및 근류형성에 미치는 영향)

  • 손상목;이장석
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.35 no.6
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    • pp.525-531
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    • 1990
  • This study was conducted to find out the effects of nitrogen, potassium application on the plant growth, grain yield and nodule formation of Winged bean. Plant height, number of branches, number of leaves, leaf area, NAR, RGR and CGR increased with basal and top dressing of nitrogen and potassium application. Especially CGR was increased significantly with the added amount of potassium. Number of nodules and nodule dry weight decreased with application of basal and top dressing nitrogen, but increased with the added amount of potassium. Green pod, tuber and grain yield increased with application of basal and top dressing nitrogen, and the added amount of potassium. There appeared positive correlation coefficient between grain yield and growth charateristics, and between grain yield and components. But there showed negative correlation coefficient among yield, number of nodules, dry weight and P$_2$O$\_$5/ contents in leaf.

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Effect of Simulated Acid Rain on the Growth and Nitrogen fixation of melilotus suaveolens (산성비가 전동싸리의 생장과 질소 고정 활성에 미치는 영향)

  • 송승달;서봉보;박재홍;박태규;정화숙;송종석;노광수;김인선
    • The Korean Journal of Ecology
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    • v.21 no.1
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    • pp.65-72
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    • 1998
  • Various effects of simulated acid rain by $HNO_3$ and $H_2SO_4$ with pH 3.2, 4.2 and 5.6 were investigated in Melilotus suaveolens Ledeb, a biennial legume dominating the disturbed and poor soil vegetations. The nitric acid treatment of pH 3.2 resulted in 121% increase of the plant height compared to that of pH 5.6 during early growth, although 17% decrease was detected with the sulfuric acids. During 14 days of treatment, leaf biomass and chlorophyll content increased 168% and 133% with pH 3.2 nitric acid rain but decreased 20% and 38%, respectively, with sulfuric acid rain. Nitrogen content in various organs was also determined after 42 days of nitric acid treatment. It increased 480% and 387% by pH 3.2 and 4.2 in leaves, 212% and 214% in stems and 247% and 249% in roots. However, the same treatment with the sulfuric acid showed a considerable reduction in this content, indicating that the nitric acid was a nitrogen source. Nodule formation assayed revealed 15% decrease with pH 3.2 in contrast to 157% increase in pH 4.2, further suggesting an enhancement effect by the additional nitrogen source. Contrary to this, the nodule formation was reduced up to 43-71% by sulfuric acid rains. Specific nitrogen fixation activities of nodules estimated at pH 3.2, 4.2 and 5.6 nitric acid rain were 36.7, 42.8 and 47.3 ${\mu}mol\;C_2H_4{\cdot}g^{-1}\;fw\;nodule{\cdot}h^{-1}$, while those of sulfuric acid rain exhibited 1 nmol $C_2H_4{\cdot}plant^{-1}{\cdot}h^{-1}$ with pH 3.2, 177 with pH 4.2 and 179 with pH 5.6, yet it increased 2, 115 and 286 respectively corresponding to the three sulfuric acid concentrations. Further implications of the simulated acied rain were also discussed in the study.

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