• 한국산림과학회지
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• 제80권1호
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• pp.1-8
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• 1991

Nodule 배양(培養) 방법(方法)을 이용(利用)하여 잡종(雜種) 포플러 양황철62-9와 이태리포플러 1호 Eco 28의 보다 진보(進步)된 재분화(再分化) 방법(方法)과 체세포(體細胞) 배(胚) 발생(發生) 방법(方法)을 얻었다. 칼루스는 2,4-D가 0.5, 2.0mg/l씩 첨가(添加)된 배지상(培地上)에 잎 조직(組織)을 치상(置床)하여 얻었고, 발달(發達)한 칼루스 조직(組織)을 액체(液體) 배지(培地)로 옮겨 세포(細胞) 현탁 배양으로 세포를 증식(增殖)했다. 현탁세포로 부터 적당한 생장(生長) 조절물질(調節物質)을 첨가(添加)하여 nodule을 생산(生産)했다. 액체 배지에서 직접(直接) 줄기를 재분화(再分化)하는 시도(試圖)는 양황철에서만 가능(可能)했다. Agar 배지(培地)에 재분화용(再分化用) 생장조절(生長調節) 물질(物質)을 첨가(添加)한 경우 상당히 많은 수(數)의 줄기 분화(分化)가 분화 되었고, 몇몇 배지에서는 체세포(體細胞) 배(胚)로 분화 했다. 이러한 nodule 배양 방법은 묘목(苗木)의 생산(生産), 체세포(體細胞) 변이(變異)의 이용(利用), 이차(二次) 산물(産物)의 생산(生産) 그리고 그밖의 생물공학적 응용을 위한 조직 배양 재료로서 그 이용성에 대하여 고찰(考察)했다.

• Journal of Microbiology and Biotechnology
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• 제16권4호
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• pp.531-537
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• 2006

Lactic acid-producing bacteria (LABs) are known to have various beneficial properties for health. However, they are generally considered to have an adverse effect on teeth, since they produce acid. Nonetheless, milk and cheese containing specific LAB strains were recently found to have an inhibitory effect on dental caries in children, with an inhibitory activity towards the growth of Streptococcus mutans suggested as the responsible mechanism. Accordingly, the current study selected a probiotic candidate for oral health and studied its inhibitory mechanism against dental caries. Twenty-two LAB species belonging to eleven genuses were screened for promoting bone nodule formation using direct microscopic examination. Only one isolate, Weissella kimchii strain PL9001, increased the bone nodule formation significantly. The addition of W. kimchii strain PL9001 to bone cells prepared from mouse calvaria increased the bone nodule formation, calcium accumulation, and activity of alkaline phosphatase (the osteoblastic marker). Moreover, W. kimchii strain PL9001 inhibited the invasion of Streptococcus mutans into bone cells, and an organic extract of the culture supernatant of W. kimchii strain PL9001 inhibited the growth of Strep. mutans. Therefore, the results suggest that W. kimchii strain PL9001 can be used as a preventive measure against dental caries. This is the first time that a LAB has been shown to promote bone nodule formation and prevent the invasion of Strep. mutans into bone cells.

• Tuberculosis and Respiratory Diseases
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• 제48권5호
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• pp.709-719
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• 2000

연구배경 : 폐결핵의의 활동성 여부를 판정하는 방법으로 객담검사에 더불어 HRCT가 유용한 것으로 알려져 있다. 이에 저자들은 객담 도말 및 배양검사의 결과와 HRCT 소견을 비교하고자 하였다. 방법 : 1996년 3월부터 1998년 9월까지 을지대학병원 호흡기내과에 내원한 활동성 폐결핵 환자를 대장으로 객담도말 및 배양검사와 HRCT를 시행하여 객담검사 결과에 따라 1군(도말(-)배양(-)군), 2군(도말(-)배양(+)군), 3군(도말(+),배양(+)군)으로 분류 하였고, 각 군에서의 HRCT소견을 비교하는 전향적 연구를 시행하였다. 결과 : 도말(+),배양(+)인 3군에서는 acinar nodule, macronodule, cavity가 각각 100%. 75%, 75%로서 도말(-)배양(-)인 1군(63%, 18%, 9%)과 도말(-)배양(+)인 2군(46%, 15%, 23%) 보다 통계적으로 유의하게 많이 관찰되었다(p<0.05). centrilobular nodule and branching structure는 3군(92%)이 1군(54%) 보다 유의하게 많았으나(p<0.05), 2군(77%)과는 통계적으로 차이가 없었다. 객담 항상균 도말 양성군은 음성군에 비해 acinar nodule(100% vs 54%), macronodule(75% vs 17%), cavity(75% vs 17%)가 통계적으로 유의하게 많이 관찰되었다(p<0.05). 결핵균 배양 양성군은 음성군에 비해서 acinar nodule(72% vs 45%), cavity(48% vs 9%)가 통계적으로 유의하게 많이 관찰되었다(p<0.05). 결론 : 임상증상과 흉부 X-선으로 활동성 폐결핵이 의심되는 환자에서 HRCT는 활동성 판정에 도움이 되며, centrilobular nodule and branching structure, acinar nodule, macronodule, cavity등의 소견은 객담 도말 검사상 음성이라도 항결핵제 투여와 함께 균 동정을 위한 유도객담 및 기관지경 검사의 필요 여부를 결정하는데 도움이 될 것으로 생각된다.

• 한국초지조사료학회지
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• 제7권3호
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• pp.128-134
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• 1987

刈取가 알팔파根瘤의 着生 및 發達과 室素固定活性에 미치는 影響을 檢討하기 위하여 圃場實驗에서는 刈取區와 非刈取區로 구분하여 時期別로 Acetylene還元法에 의하여 根瘤의 室素固定活性을 測定하였다. 養液栽培實驗에서는 地上部의 50%와 90%刈取, 花芽의 除法, 根瘤의 50%와 100% 除法등을 組合處理後 알팔파 再生과 根瘤의 着生과 發達에 미치는 影響을 檢討하였다. 1. 圃場實驗에서 根瘤의 무게는 1回刈取後 30%, 2 回刈取後 25%의 減少가 있었고 2 回刈取後 30日부터는 減少가 없었다. 2. 7月初, 8月初, 9月初에 刈取區의 specific nodule activity(SNA)는 非刈取區보다 높았으며 total nodule activity도 刈取區가 非刈取區보다 높았다. 開花期의 刈取는 根瘤의 室素固定活性 유지에 도움이 되는 것으로 나타났다. 3. 養液栽培實驗에서는 地上部의 50% 刈取는 알팔카의 再生과 根瘤의 發達에 影響이 없었으나 地上部의 90% 刈取는 再生을 느리게 하고 根瘤의 脫落을 심하게 조장하고 着生된 根瘤나 새로이 着生된 根瘤의 發達을 나쁘게 한 것으로 보아 알팔파根瘤의 室素固定活性을 높은 狀態로 維持하기 위해서는 너무 낮은 刈取를 피하는 것이 바람직할 것으로 보여진다.

• Imaging Science in Dentistry
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• 제30권3호
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• pp.189-198
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• 2000

Purpose: The study was aimed to detect the induction of apoptosis, cell cycle arrest and calcified nodule formation after irradiation on primarily cultured osteoblasts. Materials and Methods: Using rat calvarial osteoblasts, the effects of irradiation on apoptosis, cell cycle arrest, and calcified nodule formation were studied. The single irradiation of 10 and 20 Gy was done with 5.38 Gy/min dose rate using the l37Cs cell irradiator at 4th and 14th day of culture. Apoptosis induction and cell cycle arrest were assayed by the flowcytometry at 1, 2, 3, and 4 days after irradiation. The formation of calcified nodules was observed by alizarin red staining at 1, 3, 10, 14 days after irradiation at 4th day of culture, and at 1, 4, 5 days after irradiation at 14th day of culture. Results: Apoptosis was not induced by 10 or 20 Gy independent of irradiation and culture period. Irradiation did not induced G1 arrest in post-irradiated ostedblasts. After irradiation at 4th-day of culture, G2 arrest was induced but it was not statistically significant after irradiation at 14th-day of culture. In the case of irradiated cells at 4th day of culture, calcified nodules were not formed and at 14th-day of culture after irradiation, calcified nodule formation did not affected. Conclusion: Taken together, these results suggest that irradiation at the dose of 10-20 Gy would not affect apoptosis induction of osteoblasts. Cell cycle and calcified nodule formation were influenced by the level of differentiation of osteblasts.

• Journal of Periodontal and Implant Science
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• 제31권1호
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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.

• Journal of Periodontal and Implant Science
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• 제26권1호
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• pp.89-102
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• 1996

The goal of periodontal therapy is to regenerate the loss of periodontal attachment appratus. Current theories suggest the cells of the periodontium have the capacity, when appropriately triggered, to actively participate in restoring connective tissues, including mineralized tissues. This study was performed to define the hard tissue regeneration effect of periodontal ligament(PDL) cells in vitro and the effect of rate of the composition in gingival fibroblasts(GF) on the hard tissue regeneration capacity of PDL cells. For this study, Cell growth rate, alkaline phosphatase(Al.Pase) levels and the ability to produce mineralized nodules in co-culture of PDL cells and GF were examined. The results were as follows : 1. At 7 and 15 days, Cell growth of co-culture of PDL and GF(50 : 50) was greater than that of PDL cells or GF alone(P>0.05). 2. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(p<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P>0.05) 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group 1(PDL 100%), 2(PDL 70% : GF 30%), and 3(PDL 50% : GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30% GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05) From the above results, it is assumed that the co-culture of PDL cells and GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density.

• Journal of Plant Biology
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• 제36권2호
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• pp.177-182
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• 1993

The root nodules of Elaeagnus umbellata were coralloid-shape due to repeated dichotomous branching of nodule meristem. The filamentous endophyte with vesicle cluster ranging from 30 ${\mu}{\textrm}{m}$ to 60 ${\mu}{\textrm}{m}$ in diameter was present only in the cortical cells. The isolated endophytes in vitro culture showed typical Frankia morphology, consisting of highly branched hyphae ranging from 0.8 ${\mu}{\textrm}{m}$ to 1.0 ${\mu}{\textrm}{m}$ in diameter, terminal and intrahyphal sporangia varing in shape and size up to 60 ${\mu}{\textrm}{m}$ in length and laminated vesicles. Its infectivity and effectivity were confirmed by induction of nitrogen-fixing root nodules on the inoculated seedlings of two Elaeagnus species. Consequently, the isolate was confirmed as a true symbiont of Elaeagnus umbellata root nodule and named Frankia EuIK1.

• Journal of Periodontal and Implant Science
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• 제25권2호
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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.

• Journal of Periodontal and Implant Science
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• 제32권1호
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• pp.235-247
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• 2002

The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of human gingival fibroblasts. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2, and 2 mg/ml. 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 nodules was evaluated after 21 days of culture. The results were as follows : The morphology of cells on the chitosan-coated well was round or spheric. Round cells were aggregated since 6 hours of culture and showed nodule-like appearance after 24 hours of culture and did not achieved confluency at 7 days. The attachment of gingival fibroblasts was inhibited by chitosan coating with a tendency of dose dependent pattern. But, cellular activity of unit cell was higher than control. The proliferation of gingival fibroblasts was inhibited by chitosan coating at 2 mg/ml(P<0.01), while the cell proliferation at 0.02, 0.2 $mg/m{\ell}$ was comparable to the control well. Total alkaline phosphatase activity was inhibited by chitosan coating and decreased in the course of time. While ALP activity of unit cell was the highest at 2mg/ml after 4 days of culture. Finally, gingival fibroblasts produced the mineralized nodule at 2 mg/ml. In summary, the attachment, proliferation, and alkaline phosphatase activity of gingival fibroblasts were influenced differently by the concentration of coated chitosan. From this study, it could be used as the matrix of tissue engineering for gingiva without inhibition on proliferation of gingival fibroblasts using chitosan at the optimal concentration (0.02mg/ml).