• Journal of Periodontal and Implant Science
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• v.24 no.3
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• pp.581-596
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• 1994

This in vitro study was undertaken to obtain optimal tetracycline concentration that aids proliferation and spreading of human periodontal ligament cells, for clinical application in root surfaces of periodontally diseased teeth. Periodontal ligament cells used in this study were obtained from explants of periodontal ligament of 1st premolar teeth which were extracted for the purpose of orthodontic treatment. The cells were cultured in Dulbecco's Modified Eagle Medium(DMEM) supplemented with 100 U/ml penicillin, $100\;{\mu}g/ml$ streptomycin and 10% FBS at $37^{\circ}C$, 100% humidity, 5% $CO_2-95%$ air. Cells were used between the third to 4th passage. After root planing of periodontally extracted teeth, the root slabs were cut with carborundum disk. In the cell proliferation experiment, experimental groups were root planing only group, immersed groups in 25, 50, 75, 100, 150mg/ml aqueous solution of Tetracycline HCl followed by a vigorous rinse in PBS. Human PDL cells at concentration of $1{\times}10^5\;cells/ml$ were seeded in each culture well which contained root slabs and incubated for 6 hours. Then, all of the root slabs were moved into new 24 culture well and incubated 24, 48 and 72 hours. The cell counting was done by inverted phase contrast microscope after trypsinization. The following results were obtained. The cell number was increased in order root planing only group, 25, 150, 50, 75, 100mg/ml of Tetracycline HCl treated group in 24, 48 and 72 hours. The maximal cell number was obtained when the root slabs were immersed in solution with 100mg/ml of Tetracycline HCl. There were statistically significant between the root planing only group and 75, 100 mg/ml of Tetracycline HCl treated group in 24 hours, between the root planing only group and 100mg/ml of Tetracycline HCl treated group in 48 hours, between the root planing only group and 50, 75, 100mg/ml of Tetracycline HCl treated group, between 25 and 100mg/ml of Tetracycline HCl treated group in 72 hours(p<0.05). In the cell spreading experiment, after 30 minutes of incubated, in the root planing only group, the cells were generally round in shape. The cell surface was mostly covered with blebs. The cells started to attach to root surface by cytoplasmic extension in 50, 100mg/ml of Tetracycline HCl treated groups, more numerous cells attached to root surface than root planing only group. Many orifices of dentinal tubule were exposed, cells showed radially spreaded cytoplasm and unspreaded central region of the cell was covered with blebs. After 6 hours of incubation, in the root planing only group, cells showed radially spreaded cytoplasm and were attached flat appearance. In 50, 100mg/ml of Tetracycline HCl treated groups, cellular margin was concaved and cytoplasm showed elongated appearance with polarity. After 24 hours of incubation, in the root planing group, cells showed characteristic polarity. In 50, 100mg/ml of Tetracycline HCl treated groups, cells showed more elongated and spindle - like appearance.

• Korean Journal of Agricultural Science
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• v.4 no.2
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• pp.283-316
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• 1977

In an attempt to obtain a basic information to develop an effective integrated system of controlling sheath blight of rice in Korea, the transition of this disease, the variation of cultural characters and pathogenicity of the pathogen, environmental conditions affecting the disease outbreak and varietal resistance have been investigated. 1. Rice sheath blight which has been minor disease in the past was widely spread, especially since 1971. This disease has promptly spread all over the country and infected 65.2% of total rice growing area in 1976. Various factors are considered to be related to such transition of this disease. Above all, increace of application of nitrogenous fertilizer, early season and earlier cultivation of rice, introduction of more susceptible "Tongil" varieties etc. must be important factors influencing the outbreak of this disease. 2. Great variations in cultural characteristics-such as mycelial growth rate, color of the medium, amount of the aerial mycelium, shape and color of the sclerotia- and in the pathogenicity of isolates of the pathogen, Thanatephorus cucumeris Dank were observed. The optimum temperature for mycelial growth also varied with isolates, from $25^{\circ}C$ to $30^{\circ}C$. There were not necessarily any correlation between curtural characteristics and pathogenicity of isolates of Thanatephorus cucumens. 3. Mycelial grow th of isolates of Thanatephorus cucumens on the PDA medium were correlated with the air temperatures of the region where the isolates were collected. The isolates from the regions with high temperature grew well on PDA medium at $35^{\circ}C$ than those from the region with low temperature, on the other hand, the isolates from the regions with the low temperature grew well on the same medium at $12^{\circ}C$ than those from the regions with high temperature. 4. Pectin polygalacturonase (PG) and cellulase (Cx) were most active on the 3rd day after inoculation on the leaves of rice plant with Thanatephorus cucumeris, whereas pectin methylestrase (PE) was most active on the 4th day after inoculation. Relationship between the activities of PE of isolates and the strength of pathogenicity of isolates was obtained, but PG and cellulase activities were not correlated with pathogenicity of isolates. 5. The tolerence of sclerotia from in-vitro culture to low temperature varied with their water content, the dried cultural sclerotia were more tolerent than wet ones, Dried cultural sclerotia maintained almost 100% germinability for 45 days at $-20^{\circ}C$, whereas wet sclerotia lost viability at $-5^{\circ}C$. The germination ratio of the sclerotia after overwintering changed from 18% to 70% according to the water content of the test paddy fields and the ratio was low in wet paddy condition. 6. To investigate the host range of this fungi in and near paddy field, 17 weeds were inoculated with fungi. The lesions of sheath blight disease was obserbed on Sagittaria trifolia L., Echinochloa crusgalli P. Beauv., Monochoria vaginal is Presl, Polygonum Hydropiper L., Eclipta prostrata L., Digitaria sanguinalis Scapoli. 7. When the level of nitrogen applied was doubled over standard level, total nitrogen content in rice sheath increased, ami when silicate was applied, starch content in rice sheath decreased, inducing the rice plants more susceptible to sheath blight disease. Increased dressing of potash ferilizer reduced the incidence of sheat blight disease. 8. The percentage of infected stems in the early period increased more in the narrow hill plot than in the wide hill plot, but in the late period this tendency was inversed; the percentage of infected stems as well as severity in the wide hill plot increased more compared to the narrow hill plot, and the disease severity in the one plant per hill plot was also low. The number of stems in the wide hill plot was more than the number of stems in the narrow hill plot. This indicates that the microclimate, such as the relative humidity, in the narrow hill plot was more favorable for the development of this disease. 9. There was a high negative correlation between the disease severity of varieties to the sheath blight and the maturity of the varieties, that is, the early varieties were more susceptible than the late ones, and much-tillering varieties usually showed more infection than less tillering varieties. 10. No relationship was obtained between the percentage of infected stems in the early period and the severity after heading, whereas a distinct relationship was obtained between former and latter after Aug. 10.