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SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT (인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Hwang, Soon-Jeong;Choung, Pill-Hoon;Kim, Myung-Jin;Kim, Nam-Yeol;Yoo, Sang-Bae;Jahng, Jeong-Won;Kim, Hyun-Man;Kim, Joong-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.3
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

Clinical Long-term Assessment of Bioactive Glass Graft (Bioactive glass의 장기 임상적 평가)

  • Lee, Hang-Bin;Baek, Jeong-Won;Kim, Chang-Sung;Choi, Seong-Ho;Cho, Kyoo-Sung;Kim, Chong-Kwan;Chai, Choong-Kyoo
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.187-198
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    • 2002
  • The ultimate goal of periodontal therapy is the regeneration of periodontal tissue which has been lost due to destructive periodontal disease. To achieve periodontal regeneration, various kinds of methods have been investigated and developed, including guided tissue regeneration and bone graft. Bone graft can be catagorized into autografts, allografts, xenografts, bone substitutes. And materials of all types have different biological activity and the capacity for periodontal regeneration, but ideal graft material has not been developed that fits all the requirement of ideal bone graft material. Recently, bioactive glass that has been utilized in plastic surgery is being investigated for application in dental practice. But, there has not been any long-term assessment of bioactive glass when used in periodontal intrabony defects. The present study evaluates the long-term effects of bioactive glass on the periodontal regeneration in intrabony defects of human and the effect of plaqu control on long term treatment results after dividing patients into those who underwent 3-month regular check-up and those who didn't under go regular check-up The clinical effect on 74sites from 17 infrabony pockets of 11 patients were analyzed 36months after treatment. 51 sites which underwent regular check up were classified as the Follow-up group(F/U group), and 23 sites which did not undergo regular check up were classified as Non Follow-up group(Non F/U group). After comparing the probing depth, attachment loss, bone probing depth before and 36months after treatment, the following results could be concluded. 1. The changes of probing pocket depth showed a statistically significant decrease between after baseline and 36 months after treatment in F/U group(1.79${\pm}$0.68mm) and did no show astatistically significant decrease between after baseline and 36months after treatment in Non F/U group(0.61${\pm}$0.54mm) (P<0.05). 2. The changes of loss of attachment showed a statistically significant decrease between after baseline and 36 months after treatment in F/U group(1.44${\pm}$0.74mm) and did no show astatistically significant decrease between after baseline and 36months after treatment in Non F/U group(1.18${\pm}$1.54) (P<0.05). 3. The changes of bone probing depth showed a statistically significant decrease between after baseline and 36 months after treatment in both F/U(1.35${\pm}$0.28) and Non F/U group(0.78${\pm}$0.55mm) (P<0.05). The results suggest that treatment of infrabony defects with bioactive glass resulted in significan reduction of attachment loss and bone probing depth 36months after the treatment. The use of bioactive glass in infrabony defects, combined with regular check-up and proper plaque control generally shows favorable clinical results. This measn that bioactive glass could be a useful bone substitute.

The Effect of Safflower Seed Extract on the Bone Formation of Calvarial Bone Model in Sprague Dawley rat (백서 두개골 결손부에서 홍화씨 추출물의 골조직 재생 유도 효과)

  • Kim, Sung-Tae;Jhon, Gil-Ja;Lim, So-Hyoung;Cho, Kyoo-Sung;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.835-852
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    • 2000
  • The ultimate goal of periodontal therapy is the regeneration of periodontal tissue and repair of function. For more than a decade there have been many efforts to develop materials and methods of treatment to promote periodontal wound healing. Recently many efforts are concentrated on the regeneration potential of material used in oriental medicine. In some in vitro and in vivo experiments, there have been many evidences that these materials have an effect on bone regeneration. The purpose of this study was to evaluate histologically and radiologically in Sprague-Dawley rats the effects of safflower seed extracts on the regeneration of the calvarial defects surgically produced. So in this study, the critical size defects were surgically produced in the calvarial bone of 30 Sprague-Dawley rats using the 8mm trephine bur. The safflower seed extract was applied into the defect of each rat in experimental group, whereas nothing was applied into the defect of each rat in control group. Rats were sacrificed at 2, 4, 8 weeks following operation and histomorphometric and radiodensitometric analysis were performed. 1. The newly formed bone length was $102.91{\pm}22.05$, $178.29{\pm}24.40$ at 2 week in the each control, experimental group, $130.95{\pm}39.24$, $242.62{\pm}50.33$ at 4 week and $181.53{\pm}76.35$, $240.36{\pm}22.00$ at 8 week($unit,{\mu}m$). In the 2, 4 week, there were statistically significant difference between control and experimental group(P<0.05). 2. The newly formed bone area was $2962.06{\pm}1284.48$, $10648.35{\pm}1284.48$ at 2 week, $5103.25{\pm}1375.88$, $9706.78{\pm}1481.81$ at 4 week, $8046.02{\pm}818.99$, $12057.06{\pm}740.47$ at 8 week($unit,{\mu}m^2$). In every week, there were statistically significant difference between control and experimental group(P<0.05). 3. The radiopacity was $14.26{\pm}.33$, $25.47{\pm}4.33$ at 2 week, $20.06{\pm}9.07$, $26.61{\pm}2.78$ at 4 week, $22.99{\pm}3.76$, $27.29{\pm}1.54$ at 8 week(unit, %). In the 2 week, there was statistically significant difference between control and experimental group(P<0.05). In conclusion, the results of the present study suggest that safflower seed extract initially has an effect on the newly formed bone area, length and radiopacity when it is applied to the calvarial defect of Sprague - Dawley rat. Then. the material has an effect on newly formed bone area and length.

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THE EFFECTS OF POLYTETRAFLUOROETHYLENE MEMBRANE AND MILLIPORE FILTER COMBINED WITH FIBRIN ON THE REGENERATION OF PERIODONTIUM IN DOGS (Polytetrafluoroethylene membrane 및 millipore filter를 섬유소와 병용사용시 성견 치주조직 재생에 미치는 영향)

  • Joo, Sang-Don;Park, Jae-Wan;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.23 no.3
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    • pp.577-594
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    • 1993
  • The ultimate goal of periodontal therapy is the regeneration of the periodontium that have been destroyed as a result of periodontal disease. This study were done in order to determine the healing status of periodontium under Polytetrafluoroethylene and millipore fillter combined with fibrin and the effect of the guided tissue regeneration procedures were performed as follows : 1) flap operation using PTFE membrane(control group) 2) flap operation using PTFE membrane which was fixed with fibrin(experimental group 1) 3) flap operation using millipore filter which was fixed with suture(experimental group II) 4) flap operation using millipore filter which was fixed with fibrin(experimental group II) After 1, 2, 4, 8, 12 weeks, dogs were sacrificed by perfusion technique and tissue block was excised including the tooth and prepared for light microscope with H-E & Masson’s trichrome staining. The result were as follows : In control and experimental group, there is no siginificant difference on epithelial cell down growth within 1st week, but more epithelial cell downgrowth in millipore or millipore combined with fibrin group. In this experiment, there were no significant difference in new cementum and alveolar bone formation whether PTFE membrane was fixed with suture or fibrin. In control and each experimental group, bone maturation appeared in 4 weeks, bone width increased bucco-lingually in control and experimental 1 group especially. Both control group and experimental group showed mild mew cementum formation on root surface and irregular arrangement of collagen fiber at 4 weeks, that showed obvious increased cementum formation at 8 weeks, and that was observed the functional arrangement of collagen fiber between new cementum and new alveolar bone at 12 weeks.

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Physiochemical and Quality Characteristics of Young Radish (Yulmoo) Kimchi Cultivated by Organic Farming (유기농법에 의해 재배된 열무김치의 품질 및 기능적 특성)

  • Jung, Su-Jin;So, Byung-Ok;Shin, Sang-Wook;Noh, Sun-Ok;Jung, Eun-Soo;Chae, Soo-Wan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.8
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    • pp.1197-1206
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    • 2014
  • This study compared and analyzed the physiochemical and quality characteristics of young radish kimchi made with YR-FNC or YR-GC cultivated by organic farming as well as kimchi made with young radish cultivated by chemical composting/general farming (YR-Control). YR-Control showed higher contents of water and crude protein than YR-FNC and YR-GC kimchi but lower contents of crude ash, dietary fiber, and vitamin C. YR-FNC and YR-GC kimchi also showed higher contents of P and Cu than YR-Control but lower contents of N, K, Ca, Mg, Fe, and Zn. YR-Control fermentation progressed faster than that of YR-FNC or YR-GC after 24 days, and YR-Control more rapidly reached an appropriate pH. On the other hand, YR-FNC and YR-GC kimchi fermentation progressed slow and did not reach a pH level below 5.3. For period of fermentation, YR-Control and YR-FNC kimchi showed no significant difference in reducing sugar content, whereas that of YR-GC kimchi decreased significantly after 24 days of fermentation (P<0.002). YR-FNC and YR-GC kimchi showed 1.5~3 times higher total polyphenol and flavonoid contents than YR-Control at the beginning of fermentation, whereas YR-Control and YR-FNC kimchi showed a significant increase in total polyphenol and flavonoid contents during the fermentation period. By day 7 of fermentation, YR-FNC kimchi showed higher contents of lactic acid bacteria and total microbes than YR-Control. In the sensory evaluation, YR-FNC and YR-GC kimchi showed higher preference values than YR-Control. Therefore, kimchi made from young radish cultivated by organic farming has a longer freshness period than control kimchi and is characterized by excellent sensory quality, increased physiological contents, and improved beneficial health effects.

Antioxidant and Anti-inflammatory Activity of Ethanol Extract of Malus micromalus Makino in Jeju Island (제주도 자생 제주아그배 (Malus micromalus Makino) 추출물의 항산화와 항염증 활성)

  • Lee, Ju-Yeop;Kang, Min-Chul;Lee, Jung-A;Ko, Kwang-Hyo;Kim, Bong-Seok;Han, Jong-Heon;Kim, Se-Jae;Kim, Gi-Ok
    • KSBB Journal
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    • v.24 no.4
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    • pp.327-333
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    • 2009
  • The antioxidant and anti-inflammatory activities of ethanol extract of Malus micromalus were studied in vitro. Ethanol extract of M. micromalus showed scavenging effects on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and nitric oxide (NO) radicals. In addition, ethanol extract of M. micromalus inhibited the generation of superoxide anion ($O_2^-$) radical and uric acid by xanthine oxidase. We also investigated the effect of ethanol extract of M. micromalus on NO production in a lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7 cells. Ethanol extract of M. micromalus significantly inhibited NO production and this inhibitory action was not due to the cytotoxicity. The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) was markedly down-regulated by ethanol extract of M. micromalus. These results indicate that the inhibitory action of ethanol extract of M. micromalus on NO production in LPS-stimulated macropages might be due in part to abrogation of iNOS and COX-2 protein induction. Taken together, this study suggests that ethanol extract of M. micromalus could contribute to the chemoprevention and therapy of oxidative stress and inflammation.

Effect of an Acrylic Plate and SSD on Dose Profile and Depth Dose Distribution of 9 MeV Electron Beams (에너지 저하체로서 아크릴과 SSD 가 9MeV 전자선의 측방 및 깊이선량분포에 미치는 효과)

  • 강위생
    • Progress in Medical Physics
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    • v.9 no.2
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    • pp.65-71
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    • 1998
  • The aims are to evaluate the effects of an 1.0 cm acrylic plate and SSD on the dose profile and depth dose distribution of 9 MeV electron beam and to analyse adequacy for using an acrylic plate to reduce energy of electron beams. An acrylic plate of 1.0 cm thickness was used to reduce energy of 9 MeV electron beam to 7 MeV. The plate was put on an electron applicator at 65.4 cm distance from x-ray target. The size of the applicator was 10${\times}$l0cm at 100 cm SSD. For 100cm, l05cm and 110cm SSD, depth dose on beam axis and dose profiles at d$\_$max/ on two principal axes were measured using a 3D water phantom. From depth dose distributions, d$\_$max/, d$\_$85/, d$\_$50/ and R$\_$p/, surface dose, and mean energy and peak energy at surface were compared. From dose profiles flatness, penumbra width and actual field size were compared. For comparison, 9 MeV electron beams were measured. Surface dose of 7 MeV electron beams was changed from 85.5% to 82.2% increasing SSD from 100 cm to 110 cm, and except for dose buildup region, depth dose distributions were independent of SSD. Flatness of 7 MeV ranged from 4.7% to 10.4% increasing SSD, comparing 1.4% to 3.5% for 9 MeV. Penumbra width of 7 MeV ranged from 1.52 cm to 3.03 cm, comparing 1.14 cm to 1.63 cm for 9 MeV. Actual field size increased from 10.75 cm to 12.85 cm with SSD, comparing 10.32 cm to 11.46 cm for 9 MeV. Virtual SSD's of 7 and 9 MeV were respectively 49.8 cm and 88.5cm. In using energy reducer in electron therapy, depth dose distribution were independent of SSD except for buildup region as well as open field. In case of using energy reducer, increasing SSD made flatness to deteriorate more severely, penumbra width more wide, field size to increase more rapidly and virtual SSD more short comparing with original electron beam. In conclusion, it is desirable to use no energy reducer for electron beam, especially for long SSD.

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Comparison of Anisotropic Analytic Algorithm Plan and Acuros XB Plan for Lung Stereotactic Ablative Radiotherapy Using Flattening Filter-Free Beams (비편평화여과기 빔을 이용한 폐 정위절제방사선치료를 위한 AAA와 Acuros XB 계산 알고리즘의 치료계획 비교)

  • Chung, Jin-Beom;Eom, Keun-Yong;Kim, In-Ah;Kim, Jae-Sung;Lee, Jeong-Woo;Hong, Semie;Kim, Yon-Lae;Park, Byung-Moon;Kang, Sang-Won;Suh, Tae-Suk
    • Progress in Medical Physics
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    • v.25 no.4
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    • pp.210-217
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    • 2014
  • This study investigated the dosimetric effects of different dose calculation algorithm for lung stereotactic ablative radiotherapy (SABR) using flattening filter-free (FFF) beams. A total of 10 patients with lung cancer who were treated with SABR were evaluated. All treatment plans were created using an Acuros XB (AXB) of an Eclipse treatment planning system. An additional plans for comparison of different alagorithm recalcuated with anisotropic analytic algorithm (AAA) algorithm. To address both algorithms, the cumulative dose-volume histogram (DVH) was analyzed for the planning target volume (PTV) and organs at risk (OARs). Technical parameters, such as the computation times and total monitor units (MUs), were also evaluated. A comparison analysis of DVHs from these plans revealed the PTV for AXB estimated a higher maximum dose (5.2%) and lower minimum dose (4.2%) than that of the AAA. The highest dose difference observed 7.06% for the PTV $V_{105%}$. The maximum dose to the lung was also slightly larger in the AXB plans. The percentate volumes of the ipsilateral lung ($V_5$, $V_{10}$, $V_{20}$) receiving 5, 10, and 20 Gy were also larger in AXB plans than for AAA plans. However, these parameters were comparable between both AAA and AXB plans for the contralateral lung. The differences of the maximum dose for the spinal cord and heart were also small. The computation time of AXB plans was 13.7% shorter than that of AAA plans. The average MUs were 3.47% larger for AXB plans than for AAA plans. The results of this study suggest that AXB algorithm can provide advantages such as accurate dose calculations and reduced computation time in lung SABR plan using FFF beams, especially for volumetric modulated arc therapy technique.

Percutaneous Catheter Drainage of Lung Abscess (폐농양의 경피적 카테타 배농법)

  • Kim, Chang-Ho;Cha, Seoung-Ick;Han, Chun-Duk;Kim, Yeon-Jae;Lee, Yeung-Suk;Park, Jae-Yong;Jung, Tae-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.2
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    • pp.158-164
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    • 1993
  • Background: Recently, lung abscess tends to be increased in patients with underlying disease, most of whom are unsuitable for surgery when medical treatment fails. The patients with giant lung abscesses do not frequently respond to antibiotics and often have life-threatening complications. Therefore, more intensive cares are required in these patients. We studied the results and effects of percutaneous catheter drainage in these patients. Method: We performed fluoroscopy-guided percutaneous pigtail catheter (8.3 F) drainage by Seldinger technique in 9 cases of lung abscess (in 7 cases, intractable to medical treatment for an average of 8.4 days and in 2 cases, catheter drainage immediately performed due to a large cavity that was initially 10 cm in diameter). We compared 10 cases of lung abscess as control group which had receieved conventional medical treatment alone. Results: Seven of the 9 patients in study group of percutaneous drainage and 7 of the 10 patients in control group of medical treatment alone clinically improved in the average of 1.8 and 8.7 days, respectively. The mean duration of drainage was 13.2 days. There were 3 cases of death from massive hemoptysis, asphyxia of pus, and sepsis in control group, as compared with 2 cases of death from hepatic encephalopathy and sepsis in study group. The malfunctions of catheter occurred in these 2 cases, obstruction and dislodgement. But there were no significant pleuropulmonary complications of percutaneous drainage. Conclusion: Percutaneous drainage is effective and relatively safe in the management of lung abscesses refractory to medical therapy or giant lung abscesses.

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Selection of fungicides to control leaf spot of jujube (Zizyphus jujuba) trees caused by Phoma sp. (Phoma sp.에 의한 대추나무 점무늬병 방제용 살균제 선발)

  • Lee, Bong-Hun;Lim, Tae-Heon;Cha, Byeong-Jin
    • The Korean Journal of Pesticide Science
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    • v.4 no.3
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    • pp.40-46
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    • 2000
  • To select the effective fungicides for the control of leaf spot disease of jujube tree (Zizyphus jujuba) caused by Phoma sp., inhibitory effects of 26 fungicides for mycelial growth were investigated at $250{\mu}g\;a.i./m{\ell}$. In the test, eight fungicides were selected and minimum inhibitory concentration (MIC) for mycelial growth and an inhibitory effect for spore germination were investigated. Among the fungicides, myclobutanil, hexaconazole, and triflumizole were excluded in control effect tests because of their relatively high MICs. MICs were ranged $10-50{\mu}g\;a.i./m{\ell}$ for benomyl, carbendazim + kasugamycin (CK), and thiophanate-methyl. triflumizole (TT), and $50-250{\mu}g\;a.i./m{\ell}$ for iprodione + propineb (IT) and iminoctadine-triacelate (IT). However, benomyl and IP showed very low inhibitory effect on conidial germination. When the fungicides were sprayed on the seedlings before the leaves were inoculated with conidial suspension of Phoma sp., the protective values of CK and TT were around 70% at 1,000 ppm and around 90% at 2,000 ppm. The protective values were around 70% at 2,000 ppm (benomyl), 4,000 ppm (IP), and 8,000 ppm (IT). When the fungicides were sprayed after inoculation, benomyl showed the highest curative values of over 90% at 1,000 ppm and the values of CK and TT ranged $70{\sim}80%$ at 1,000 ppm. However, IP and IT had little or no effect on therapy of the disease. IT caused necrotic phytotoxicity on the leaves of jujube seedlings. As results, the best fungicides for the protection of jujube trees from leaf spot disease were CK (2,000 ppm) and TT (2,000 ppm) and for the remedy of the tree, benomyl (1,000 ppm) was the best. Therefore, alternate application of benomyl and CK or TT will be effective in the disease control.

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