• Title/Summary/Keyword: 분주(分註)

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Variation in Ploidy Level of Rice Plants Derived from Anther Culture (벼 약배양에서 유기된 식물체의 배수성)

  • Sohn, Jae-Keun;Lee, Su-Kwan;Oh, Byong-Geun;Park, Rae-Kyong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.29 no.4
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    • pp.328-333
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    • 1984
  • Variation in ploidy level of regenerated plants from rice anthers and effective diploidization methods of haploid plants were studied to obtain basic information in rice breeding through anther culture. In a total of 574 plants derived from anther culture using 14F$_1$ hybrids as materials, there were 49.7% haploids, 48.6% diploids and 1.7% polyploids, respectively. The frequency of haploids in Japonica/Indica crosses was 60.6%, and that of Japonica/Japonica crosses was 43.0% in average. Inclusion of 2.4-D or NAA as phytohormone may increase the frequency of haploids, but kinetin may increase the frequency of diploids. The rate of auto-diploidization by tiller separation of haploid plants showed 8.2% in average. The rate of diploidization by leaf-sheath injection of colchicine showed 18.8% in average. Morphological characters of haploids plants showed that 64.6% in culm length, 63.4% in panicle length, 68% in flag leaf length, and 74.4% in flag leaf width compared to diploid plants. These apparent morphological differences will contribute to identify the ploidy of plants derived from rice anther culture.

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Evaluation of Insecticidal Efficacy of Six Eco-friendly Agricultural Materials and Metarhizium anisopliae against Ramulus mikado (대벌레(Ramulus mikado)에 대한 유기농업자재 6종과 녹강균(Metarhizium anisopliae)의 살충 효과 평가)

  • Jong-Kook Jung;Bok-Nam Jung;Cha Young Lee;Keonhee E. Kim;Junheon Kim;Young Su Lee;Ji-Hyun Park
    • Journal of Korean Society of Forest Science
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    • v.112 no.1
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    • pp.117-125
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    • 2023
  • Outbreaks of Ramulus mikado (Insecta: Phasmatodea: Phasmatidae) in the hilly areas of Mt. Bongsan, Mt. Cheonggye, and elsewhere in Seoul and Gyeonggi occurred from 2020 to 2021, causing serious defoliation. We evaluated the insecticidal effects of six eco-friendly organic materials and the insect-pathogenic fungus Metarhizium anisopliae against R. mikado. The fungus was isolated from naturally dead bodies of R. mikado in forest ecosystems. The results revealed that three eco-friendly organic materials containing azadirachtin or geraniol as active ingredients showed high mortality in the range of 85.2%-100%, which were rates similar to that of the chemical insecticide fenitrothion emulsifiable concentrate. All R. mikado adults that were sprayed with a conidial suspension of M. anisopliae at different concentrations were killed within a few days. In conclusion, three eco-friendly organic materials and M. anisopliae could be good alternatives to chemical insecticides.

Amino Acid Concentrations in the Blastocoelic Fluid of In Vitro-Produced Bovine Blastocysts (체외생산된 소 배반포강 내의 아미노산 농도)

  • Jung, Y.G.;Wei, H.;Yoon, J.T.;Fukui, Y.
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.251-259
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    • 1998
  • Concentrations of free amino acids in the BF of IVP bovine BL and HBL were examined in this study. The embryos derived from IVF oocytes were cultured in a SOFM containing BSA, EAA and NEAA. BF was aspirated from BL (180 h of age after insemination) and HBL (216 h of age after insemination), and introduced into drops of SOFM (30$\mu$l/drop) containing PVA through micromanipulation. The medium containing BF was then subjected to measurement of 20 amino acids by an automatic amino acid analyzer. The concentrations of isoleucine, leucine and methionine were higher (p〈0.05) in the BF from HBL than from BL, and no difference was found in aspartate or glutamate concentrations between BL and HBL, while threonine, alanine (p〈0.01) and the rest of the amino acids (p〈0.001) were significantly higher in the BF from HBL than from BL. Cystine was not found in either BL or HBL. A high concentration of glutamine was found in the BF from both BL and HBL, although it was not added to the culture medium. These results indicate that bovine BF contains several EAA (methionine in BL and isoleucine, leucine and methionine in HBL) and NEAA (alanine, glutamate, glycine, proline, serine and aspartate in BL, and glutamate and aspartate in HBL), and there is significant differences in the amino acid concentration in the BF between BL and HBL derived by WP.

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Effect of Rumex acetosella Extract on Germination and Growth of Festuca arundinacea

  • Yosep Kang;Но-Jun Gam;Eun-Jung Park;Bo-Ram Choi;Ki-Yong Kim;Sang-Mo Kang; In-Jung Lee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.112-112
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    • 2022
  • 외래잡초인 애기수영(Rumex acetosella)는 생태계 교란종으로 생물다양성을 감소시키고, 우리나라 목초지에 우점하고 있어 큰 문제를 야기한다. 애기수영의 경우, 살초효과 및 제초활성물질인 chrysophanic acid와 catechd이 밝혀져 있지만, 톨페스큐(Festuca arundinacea)에 대한 살초 효과 연구는 미비하다. 이에 본 연구는 톨페스큐 종자에 대해 애기수영 MeOH 추출물을 처리한 Seed bioassay를 진행해 IC50 값을 구하고, 톨페스큐에 애기수영 MeOH 추출물을 경엽처리를 진행한 후 생육조사를 진행했다. Seed bioassay의 경우, petri dish 위에 톨페스큐 종자 20개가 치상하고, 애기수영 지상부 추출물과 지하부 추출물을 각각 20,000 mg L-1, 10,000 mg L-1, 5,000 mg L-1, 2,500 mg L-1 농도로 serial dilution 하여 1mL씩 분주한 뒤, 일주일 뒤에 발아한 종자에 대해 생체중을 조사하고 Prizm 프로그램을 이용해 IC50을 구하였다. 경엽처리의 경우, 톨페스큐 종자 파종4주 뒤에 IC50값이 더 낮았던 지상부 추출물을 100,000 mg L-1, 50,000 mg L-1, 25,000 mg L-1, 12,500 mg L-1, 6,250 mg L-1 농도로 serial dilution 한 뒤 5mL씩 일주일 간격으로 3회 경엽처리를 진행하였고, 마지막 처리 일주일 뒤 초장, 근장, 생체중, 건물중을 조사하였다. Seed bioassay 결과, 애기수영 지하부 추출물에 대한 톨페스큐의 IC50값은 3274가 나왔고, 애기수영 지상부 추출물에 대한 톨페스큐의 IC50값은 2728가 나왔다. Seed bioassay 결과를 바탕으로 효과적 이 었던 지상부 추출물을 이용해 톨페스큐 경엽처리를 진행하였다. 애기수영 지상부 추출물 경엽처리 결과, 톨페스큐 초장과 생체중이 추출물 처리량이 높아짐에 따라 낮아졌으며, 100,000 mg L-1 처리구는 Control과 비교해 유의적으로 감소하였고, 처리량이 높아짐에 따라 근장이 감소했지만, 유의적인 차이는 없었다. 그리고 건물중은 100,000 mg L-1 처리구가 Control, 12,500 mg L-1, 6,250 mg L-1 처리구와 비교해 유의적으로 낮았다.

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Analysis of Variation for Parallel Test between Reagent Lots in in-vitro Laboratory of Nuclear Medicine Department (핵의학 체외검사실에서 시약 lot간 parallel test 시 변이 분석)

  • Chae, Hong Joo;Cheon, Jun Hong;Lee, Sun Ho;Yoo, So Yeon;Yoo, Seon Hee;Park, Ji Hye;Lim, Soo Yeon
    • The Korean Journal of Nuclear Medicine Technology
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    • v.23 no.2
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    • pp.51-58
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    • 2019
  • Purpose In in-vitro laboratories of nuclear medicine department, when the reagent lot or reagent lot changes Comparability test or parallel test is performed to determine whether the results between lots are reliable. The most commonly used standard domestic laboratories is to obtain %difference from the difference in results between two lots of reagents, and then many laboratories are set the standard to less than 20% at low concentrations and less than 10% at medium and high concentrations. If the range is deviated from the standard, the test is considered failed and it is repeated until the result falls within the standard range. In this study, several tests are selected that are performed in nuclear medicine in-vitro laboratories to analyze parallel test results and to establish criteria for customized percent difference for each test. Materials and Methods From January to November 2018, the result of parallel test for reagent lot change is analyzed for 7 items including thyroid-stimulating hormone (TSH), free thyroxine (FT4), carcinoembryonic antigen (CEA), CA-125, prostate-specific antigen (PSA), HBs-Ab and Insulin. The RIA-MAT 280 system which adopted the principle of IRMA is used for TSH, FT4, CEA, CA-125 and PSA. TECAN automated dispensing equipment and GAMMA-10 is used to measure insulin test. For the test of HBs-Ab, HAMILTON automated dispensing equipment and Cobra Gamma ray measuring instrument are used. Separate reagent, customized calibrator and quality control materials are used in this experiment. Results 1. TSH [%diffrence Max / Mean / Median] (P-value by t-test > 0.05) C-1(low concentration) [14.8 / 4.4 / 3.7 / 0.0 ] C-2(middle concentration) [10.1 / 4.2 / 3.7 / 0.0] 2. FT4 [%diffrence Max / Mean / Median] (P-value by t-test > 0.05) C-1(low concentration) [10.0 / 4.2 / 3.9 / 0.0] C-2(high concentration) [9.6 / 3.3 / 3.1 / 0.0 ] 3. CA-125 [%diffrence Max / Mean / Median] (P-value by t-test > 0.05) C-1(middle concentration) [9.6 / 4.3 / 4.3 / 0.3] C-2(high concentration) [6.5 / 3.5 / 4.3 / 0.4] 4. CEA [%diffrence Max / Mean / median] (P-value by t-test > 0.05) C-1(low concentration) [9.8 / 4.2 / 3.0 / 0.0] C-2(middle concentration) [8.7 / 3.7 / 2.3 / 0.3] 5. PSA [%diffrence Max / Mean / Median] (P-value by t-test > 0.05) C-1(low concentration) [15.4 / 7.6 / 8.2 / 0.0] C-2(middle concentration) [8.8 / 4.5 / 4.8 / 0.9] 6. HBs-Ab [%diffrence Max / Mean / Median] (P-value by t-test > 0.05) C-1(middle concentration) [9.6 / 3.7 / 2.7 / 0.2] C-2(high concentration) [8.9 / 4.1 / 3.6 / 0.3] 7. Insulin [%diffrence Max / Mean / Median] (P-value by t-test > 0.05) C-1(middle concentration) [8.7 / 3.1 / 2.4 / 0.9] C-2(high concentration) [8.3 / 3.2 / 1.5 / 0.1] In some low concentration measurements, the percent difference is found above 10 to nearly 15 percent in result of target value calculated at a lower concentration. In addition, when the value is measured after Standard level 6, which is the highest value of reagents in the dispensing sequence, the result would have been affected by a hook effect. Overall, there was no significant difference in lot change of quality control material (p-value>0.05). Conclusion Variations between reagent lots are not large in immunoradiometric assays. It is likely that this is due to the selection of items that have relatively high detection rate in the immunoradiometric method and several remeasurements. In most test results, the difference was less than 10 percent, which was within the standard range. TSH control level 1 and PSA control level 1, which have low concentration target value, exceeded 10 percent more than twice, but it did not result in a value that was near 20 percent. As a result, it is required to perform a longer period of observation for more homogenized average results and to obtain laboratory-specific acceptance criteria for each item. Also, it is advised to study observations considering various variables.

Effects of Terpenoids-Rich Plant Extracts on Ruminal-fermentation and Methane Production (Terpenoid 함유 식물 추출물의 첨가가 반추위 발효와 메탄 발생에 미치는 영향)

  • Hwang, Hee-Soon;Ha, Dong-Uk;Lee, Su-Kyoung;Lee, Il-Dong;Lee, Shin-Ja;Lee, Sung-Sill
    • Korean Journal of Organic Agriculture
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    • v.21 no.4
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    • pp.629-646
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    • 2013
  • This study was conducted to investigate effects of terpenoids-rich plant extracts (TRPE) on the in vitro ruminal fermentation characteristics and methane production. The ruminal fluid was collected from a cannulated Hanwoo cow fed concentrate and timothy in the ratio of 6 to 4. The TRPE as Mint (Mentha arvensis var. piperascens), Pine (Pinus densiflora), Japan cedar (Cryptomeria japonica), Sichuan pepper (Zanthoxylum piperitum), Hinoki cypress (Chamaecyparis obtuse) and Japanese black pine (Pinus thunbergii) were used in this study. The 15 mL of mixture, contains McDougall buffer and rumen fluid in the ratio of 2 to 1. The mixture was dispensed anaerobically 50 mL serum bottles and it is contained 0.3 g timothy substrate and 5% TRPE. The bottles were incubated at $39^{\circ}C$ for 3, 6, 9, 12, 24, 48 and 72 hours. The pH value decrease by increased incubation times and the pH values at all times were significantly (p<0.05) higher in treatments than in control. The digestibility of dry matter at 3 hours was significantly (p<0.05) higher in mint treatment than in control. Productions of total gas and carbon dioxide at before 12 hours was significantly lower (p<0.05) in treatments than in control. The methane production at 24 hours was significantly (p<0.05) lower in treatments than in control. The concentrations of acetic acid and propionic acid at 24 hours were significantly higher (p<0.05) in mint and pine treatments than in control. In conclusion, the terpenoid-rich plant extracts were shown to decreased methane emission and without adversely affected ruminal fermentation. Therefore, the terpenoid-rich plant extracts as mint and pine were shown to decreased methane production and it has potential possibility for ruminal fermentations.

The Optimal Condition of Performing MTT Assay for the Determination of Radiation Sensitivity (방사선 감수성 측정법으로서 MTT 법 시행 시의 최적 조건에 대한 연구)

  • Hong, Se-Mie;Kim, Il-Han
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.163-170
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    • 2001
  • Purpose : The measurement of radiation survival using a clonogenic assay, the established standard, can be difficult and time consuming. In this study, We have used the MTT assay, based on the reduction of a tetrazolium salt to a purple formazan precipitate by living cells, as a substitution for clonogenic assay and have examined the optimal condition for performing this assay in determination of radiation sensitivity. Materials and Methods : Four human cancer cell lines - PCI-1, SNU-1066, NCI-H630 and RKO cells have been used. For each cell line, a clonogenic assay and a MTT assay using Premix WST-1 solution, which is one of the tetrazolium salts and does not require washing or solubilization of the precipitate were carried out after irradiation of 0, 2, 4, 6, 8, 10 Gy. For clonogenic assay, cells in $25\;cm^2$ flasks were irradiated after overnight incubation and the resultant colonies containing more than 50 cells were scored after culturing the cells for $10\~14$ days. For MTT assay, the relationship between absorbance and cell number, optimal seeding cell number, and optimal timing of assay was determined. Then, MTT assay was performed when the irradiated cells had regained exponential growth or when the non-irradiated cells had undergone four or more doubling times. Results : There was minimal variation in the values gained from these two methods with the standard deviation generally less than $5\%$, and there were no statistically significant differences between two methods according to t-test in low radiation dose (below 6 Gy). The regression analyses showed high linear correlation with the $R^2$ value of $0.975\~0.992$ between data from the two different methods. The optimal cell numbers for MTT assay were found to be dependent on plating efficiency of used cell line. Less than 300 cells/well were appropriate for cells with high plating efficiency (more than $30\%$). For cells with low plating efficiency (less than $30\%$), 500 cells/well or more were appropriate for assay. The optimal time for MTT assay was after 6 doubling times for the results compatible with those of clonogenic assay, at least after 4 doubling times was required for valid results. In consideration of practical limits of assay (12 days, in this study) cells with doubling time more than 3 days were inappropriate for application. Conclusion : In conclusion, it is found that MTT assay can successfully replace clonogenic assay of tested cancer cell lines after irradiation only if MTT assay was undertaken with optimal assay conditions that included plating efficiency of each cell line and doubling time at least.

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The Lymphocyte Dependent Bactericidal Assay of Human Monocyte and Alveolar Macrophage for Mycobacteria (마이코박테리아에 대한 인체 말초혈액 단핵구와 폐포대식세포의 림프구 의존적 살해능에 관한 연구)

  • Cheon, Seon-Hee;Lee, You-Hyun;Lee, Jong-Soo;Bae, Ki-Sun;Shin, Sue-Yeon
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.1
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    • pp.5-16
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    • 2002
  • Background : Though mononuclear phagocytes serve as the final effectors in killing intracellular Mycobacterium tuberculosis, the bacilli readily survive in the intracellular environment of resting cells. The mechanisms through which cellular activation results in the intracellular killing is unclear. In this study, we sought to explore an in vitro model of a low-level infection of human mononuclear phagocytes with MAC and $H_{37}Ra$ and determine the extent of the lymphocyte dependent cytotoxicity of human monocytes and alveolar macrophages. Materials and Methods : The peripheral monocytes were prepared using the Ficoll gradient method from PPD positive healthy people and tuberculosis patients. The alveolar macrophages were prepared from PPD positive healthy people via a bronchoalveolar lavage. The human mononuclear phagocytes were infected at a low infection rate (bacilli:phagocyte 1:10) with MAC(Mycobacterium avium) and Mycobacterium tuberculosis $H_{37}Ra$. Non-adherent cells(lymphocyte) were added at a 10:1 ratio. After 1,4, and 7 days culture in $37^{\circ}C$, 5% CO2 incubator, the cells were harvested and inoculated in a 7H10/OADC agar plate for the CFU assay. The bacilli were calculated with the CFU/$1{\times}10^6$ of the cells and the cytotoxicity was expressed as the log killing ratio. Results : The intracellular killing of MAC and $H_{37}Ra$ within the monocyte was greater in patients with tuberculosis compared to the PPD positive controls (p<0.05). Intracellular killing of MAC and $H_{37}Ra$ within the alveolar macrophage appeared to be greater than that within the monocytes of the PPD positive controls. There was significant lymphocyte dependent inhibition of intracellular growth of the mycobacteria within the monocytes in both the controls and tuberculosis patients and within the macrophages in the controls(p<0.05). There was no specific difference in the virulence between the MAC and the $H_{37}Ra$. Conclusion : This study is an in vitro model of a low-level infection with MAC and $H_{37}Ra$ of human mononuclear phagocytes. The intracellular cytotoxicity of the mycobacteria within the phagocytic cells was significantly lymphocyte dependent. During the 7 days culture after the intracellular phagocytosis, the actual confinement of the mycobacteria was observed within the monocytes of tuberculosis patients and the alveolar macrophages of the controls as in the case of adding lymphocytes.

Study on the Effect of Blending Ratios on the Antibacterial Activities of Chitosan/Gelatin Blend Solutions (혼합비율에 따른 키토산/젤라틴 혼합용액의 항균활성에 관한 연구)

  • Kim, Byung-Ho;Park, Jang-Woo;Hong, Ji-Hyang
    • Korean Journal of Food Science and Technology
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    • v.37 no.3
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    • pp.405-411
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    • 2005
  • Chitosan, second largest biomass after cellulose on earth, has potential for use as functional food package due to its antibacterial activity. However, due to high melting temperature of chitosan, chitosan films have been made by casting method. Because gelatin has relatively low molting temperature depending upon amount of plasticizer added, it was added to chitosan to produce commercially feasible film. The objective of the current study was to determine optimum blend ratio and amount of chitosan/gelatin blend solutions against antibacterial activities for extruder resin. Gram-positive bacteria (Bacillus cereus ATCC 14579 and Listeria monocytogenes ATCC 15313) and -negative bacteria (Escherichia coli ATCC 25922 and Salmonella enteritidis IFO 3313) were used. Paper (8 mm) diffusion and optical density methods were used to evaluate effect of different blending ratio solutions on the inhibition of bacterial growth. Measured clear none size ranged from 8 mm to 18.07 mm in paper diffusion test. For B. cereus, E. coli, and S. enteritidis, addition of $50\;{\mu}L$ blend solution (chitosan/gelatin = 2/8: 0.3 mg) resulted in clear zone on paper disc. In L. monocytogenes, inhibition effect was observed with 0.6 mg chitosan (chitosan/gelatin=4/6). Minimum inhibitory concentration (MIC) values of B. cerues, L. monocytogenes, E. coli, and S. enteritidis with addition of chitosan were 0.1461, 0.2419, 0.0980, and 0.0490 mg/mL, respectively, These results indicate possibility of producing commercially feasible film with addition of optimum chitosan/gelatin amount.

The Effect of the IGF-I treated Gingival and Periodontal Ligament Fibroblast on Osteoblasts (IGF-I으로 처리한 치은 및 치주인대 섬유모세포가 골모세포에 미치는 영향)

  • Kim, Mi-Jeong;Yang, Won-Sik
    • The korean journal of orthodontics
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    • v.31 no.6 s.89
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    • pp.589-600
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    • 2001
  • Insulin-like growth factor I (IGF-I) has the local tissue regulating actions. In bone, IGF-I increases the replication of osteoblastic lineage, probably preosteoblasts, and enhances osteoblastic collagen synthesis and matrix composition rates. The purpose of this study was to investigate the local regulatory effect of IGF-I on periodontium totally, both in an autocrine and paracrine manner. To examine the effect of IGF-I directly on osteoblast (OB) of test rats, and indirectlv on OB via periodontal ligament fibroblast (PDLF), and the effect of gingival fibroblast (GF) on OB via cellular paracrine manner for the understanding of humoral action of adjacent tissue, GF and PDLF were obtained from male Sprague-Dawley rats of six to eight weeks of age. OB was obtained iron frontal and parietal calvarial bone of Sprague-Dawley 21day-old-fetus. After each tell was Incubated 24 hours, for collecting conditioned medium, different concentrations of IGF-I (1,10,100 ng/ml,1ml/well) was adding in the GF, PDLF cells, and the supernatant from these cultures was put into the primary OB culture with $1{\times}10^4$cell/ml/well. The experimental group was divided into six groups control OB, IGF-I treated OB, OB culture with conditioned medium from PDLF, OB culture with conditioned medium from IGF-I treated PDLF, OB culture with conditioned medium from GF, OB culture with conditioned medium from IGF-I treated GF. After final IGF-I treatment, OB was Incubated for 24 hours, and alkaline phosphatase activity assay, BMP expression, cell proliferation measurement using MTT assay, total protein measurement, Collagen synthesis assay using western blot, and examination of bone nodule synthesis were done. Alkaline phosphatase expressions were increased in the group of PDLF-IGF-I supernatant treatment. Direct IGF-I treatment with concentrations of 100ng/m1 showed increased viable tell number measured by MTT assay. And IGF-I treatment did not increase total protein amount. The entire experimental group showed BMP2, 4 expression in western blot, and there was no significant difference between control and experimental groups. These results suggested that supernatant from PDLF effects on increasing cellular activities of OB regardless of IGF-I, and at high concentration, IGF-I increases OB tell proliferation.

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