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Formulation of Bacillus amyloliquefaciens A-2 and Its Efficacy to Control Tomato Leaf Mold Caused by Fulvia fulva (길항세균 Bacillus amyloliquefaciens A-2를 이용한 토마토 잎곰팡이병 방제용 미생물 제제)

  • Kong, Hyun-Gi;Chun, Ock-Joo;Choi, Ki-Hyuck;Lee, Kwang-Youll;Baek, Joung-Woo;Kim, Hyun-Ju;Murugaiyan, Senthilkumar;Moon, Byung-Ju;Lee, Seon-Woo
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.27-34
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    • 2010
  • This study was performed to develop a formulation using an antagonistic bacterium Bacillus amyloliquefaciens A-2 to control tomato leaf mold caused by Fulvia fulva. B. amyloliquefaciens A-2 was grown in a medium with rice oil and mixed with various carrier and additives. One of the formulations, A2-MP, showed the best disease control value among the tested formulations. The disease control value of A2-MP at 100-fold and 500-fold diluted treatment was not significantly different from that of chemical fungicide triflumizole in a growth chamber. Although disease control effect was decreased by serial diluted treatment of the prepared A2-MP, 1,000-fold diluted treatment of A2-MP still showed high disease control value of 72.0%. For the green house experiments, the disease control values of A2-MP was indicated as 79.4% which is similar to that of chemical fungicide, triflumizole showing 79.6%. When the disease control activity of the formulation A2-MP was compared in tomato production conditions, disease control values of 100-fold diluted A2-MP and 3,000 fold diluted triflumizole exhibited 60%, 81.6%, respectively. The disease control efficiency by A-2MP was 73% of the disease control value of chemical fungicide. The formulation A-2MP maintained the stable bacterial viability and disease control activity when stored at $4^{\circ}C$. This result suggested that A-2MP develped from B. amyloliquefaciens A-2 could be used to control tomato leaf mold.

Effect of Acupuncture on the Expressions of Neuropeptide Y and Leptin Receptor in the Hypothalamus of Food-deprived Rats (침치료가 굶긴 쥐 시상하부에서 neuropeptide Y(NPY)와 leptin receptor(LR)의 발현에 미치는 영향에 대한 실험적 연구)

  • Kim, Mi-A;Jung, Woo-Sang;Moon, Sang-Kwan;Kim, Young-Suk;Kim, Chang-Ju;Cho, Ki-Ho
    • The Journal of Internal Korean Medicine
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    • v.34 no.2
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    • pp.113-121
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    • 2013
  • Objectives : This study aimed to find out whether acupuncture at various acupoints shows any effects on appetite by regulating neurotransmitters in the hypothalamus through the expression of NPY and LR in the PVN via immunohistochemistry. Methods : Male Sprague-Dawley rats were divided into eight groups of five mice each. The rats in the acupuncture groups were treated with acupuncture at respective acupoints, twice per a day for 3 days. The animals were sacrificed 72 h after commencement of the experiment, the brains being dissected into serial coronal sections. Expressions of neuropeptide Y and leptin receptor in the hypothalamus were assessed by immunohistochemistry. Results : NPY expression in the PVN was enhanced and LR expression in the PVN was decreased by food deprivation. NPY immunoreactivity in the PVN of the food-derived rats was decreased by acupuncture at the auricular acupoint, Zusanli-acupoint, a and non-acupoint. However, acupuncture at the auricular acupoint showed most potent suppressing effect on NPY expression in the PVN of food-deprived rats. LR expression in the PVN decreased following food deprivation, and auricular acupuncture increased LR expression in the PVN of food-deprived rats. In normal conditions (fed state), LR expression in the PVN was not changed by acupuncture treatment at several sites. Conclusions : From this study, we have shown that acupuncture at the auricular acupoint exerts the most potent appetite suppressing effect on the food restriction state.

Effects of the Dosing Regimen of Tissue-type Plasminogen Activator on Blood Coagulation System in Experimental Pulmonary Embolism (실험적 폐색전증에서 조직형플라스미노겐활성체의 투여방법이 혈액응고기전에 미치는 영향)

  • Chung, Hee-Soon;Kim, Ho-Joong;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.5
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    • pp.474-482
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    • 1993
  • Background: As a physiologic plasminogen activator, tissue-type plasminogen activator (t-PA) could induce effective thrombolysis in massive pulmonary embolism, without the risk of systemic hemorrhage. However, therapeutic doses of t-PA has been associated with systemic lytic state, and fibrin selectivity may be influenced by the dosing regimen of t-PA. To investigate the effects of duration of t-PA infusion on blood coagulation system, we performed this study. Method: In a canine model of pulmonary embolism, which was induced by injection of autologous blood clots, we administered equal doses of t-PA (1 mg/kg) over 15 minutes in $t-PA_{15}$ group, over 180 minutes in $t-PA_{180}$ group, and only saline in control group. Then serial blood samplings were made to check complete blood count, prothrombin time, activated partial thromboplastin time, thrombin time, fibrin, plasminogen, ${\alpha}_2$-antiplasmin, coagulation factor V and VIII, and fibrin(ogen) degradation products. Results: 1) In all 3 groups, complete blood count showed same changes. Hemoglobin, hematocrit and platelet count decreased, but WBC count increased. 2) Prothrombin time, activated partial thromboplastin time, and thrombin time were prolonged during 15-60 minutes after t-PA administration in $t-PA_{15}$ group, and from 30 minutes through 180 minutes after administration in $t-PA_{180}$ gorup. 3) Fibrin, ${\alpha}_2$-antiplasmin, and cogulation factor V and VIII decreased in both $t-PA_{15}$ and $t-PA_{180}$ group, but returned to basal levels earlier in $t-PA_{15}$ group. 4) Fibrin(ogen) degradation products increased after pulmonary embolism in all groups, and further increased in both $t-PA_{15}$ and $t-PA_{180}$ groups after t-PA infusion. But more pronounced increment was noted in $t-PA_{180}$ gorup. Conclusion: In pulmonary embolism, the shorter (15 minutes) infusion of t-PA would have less risk of systemic hemorrhage than the longer (180 minutes) infusion when the doses is equal. And, this suggests that manipulating the duration of t-PA infusion can reduce the risk of major bleeding.

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The Role of Chest CT Scans in the Management of Empyema (농흉에서 전산화 단층촬영의 의의)

  • Heo, Jeong-Suk;Kwun, Oh-Yong;Sohn, Jeong-Ho;Choi, Won-Il;Hwang, Jae-Seok;Han, Seung-Beom;Jeon, Young-June;Kim, Jung-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.4
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    • pp.397-404
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    • 1994
  • Background: To decide the optimal antibiotics and application of chest tube, examination of pleural fluid is fundamental in the management of empyema. Some criteria for drainage of pleural fluid have been recommended but some controversies have been suggested. Recently, newer radiologic methods including ultrasound and computed tomography scanning, have been applied to the diagnosis and management of pleural effusions. We undertook a retrospective analysis of 30 patients with pleural effusion who had CT scans of the chest in order to apply the criteria of Light et al retrospectively to patients with loculation and to correlate the radiologic appearance of pleural effusions with pleural fluid chemistry. Method: We analyzed the records of 30 out of 147 patients with pleural effusion undergoing chest CT scans. Results: 1) Six of the pleural fluid cultures yielded gram negative organisms and three anaerobic bacterias and one Staphylococcus aureus and one non-hemolytic Streptococci. No organism was cultured in ninteen cases(63.0%). 2) The reasons for taking chest CT scans were to rule out malignancy or parenchymal lung disease(46.7%), poor response to antibiotics(40.0%), hard to aspirate pleural fluid(10.0%) and to decide the site for chest tube insertion(3.3%). 3) There was no significant correlations between ATS stages and loculation but there was a tendency to loculate in stage III. 4) There was a significant inverse relationship between the level of pH and loculation(p<0.05) but there appeared to be no relationship between pleural fluid, LDH, glucose, protein, loculation and pleural thickening. 5) In 12 out of 30, therapeutic measures were changed according to the chest CT scan findings. Conclusion: We were unable to identify any correlations between the plerual fluid chemistry, ATS stages and loculations except pH, and we suggest that tube thoracotomy should be individualized according to the clinical judgement and serial observation. All patients with empyema do not need a chest CT scan but a CT scan can provide determination of loculation, guiding and assessing therapy which should decrease morbidity and hospital stay.

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Study on Tumor Control Probability and Normal Tissue Complication Probability in 3D Conformal Radiotherapy (방사선 입체조형치료에 대한 종양치유확율과 정상조직손상확율에 관한 연구)

  • 추성실
    • Progress in Medical Physics
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    • v.9 no.4
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    • pp.227-245
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    • 1998
  • A most appropriate model of 3-D conformal radiotherapy has been induced by clinical evaluation and animal study, and therapeutic gains were evaluated by numerical equation of tumor control probability(TCP) and normal tissue complication probability (NTCP). The radiation dose to the tumor and the adjacent normal organs was accurately evaluated and compared using the dose volume histogram(DVH). The TCP and NTCP was derived from the distribution of given dosage and irradiated volume, and these numbers were used as the biological index for the assessment of the treatment effects. Ten patients with liver disease have been evaluated and 3 dogs were sacrificed for this study. Based on the 3-D images of the tumor and adjacent organs, the optimum radiation dose and the projection direction which could maximize the radiation effect while minimizing the effects to the adjacent organs could be decided. 3). The most effective collimation for the normal adjacent organs was made through the beams eye view with the use of multileaf collimator. When the dose was increased from 50Gy to 70Gy, the TCP for the conventional 2-port radiation and the 5-port multidimensional therapy was 0.982 and 0.995 respectively, while the NTCP was 0.725 and 0.142 respectively, suggesting that the 3-D conformal radiotherapy might be the appropriate therapy to apply sufficient radiation dose to the tumor while minimizing the damages to the normal areas of the liver. Positive correlation was observed between the NTCP and the actual complication of the normal liver in the animal study. The present study suggest that the use of 3-D conformal radiotherapy and the application of the mathematical models of TCP and NTCP may provide the improvements in the treatment of hepatoma with enhanced results.

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Ototoxicity in children receiving cisplatin chemotherapy (Cisplatin을 포함한 항암치료를 받은 소아에서 이독성)

  • Jang, Hee Jin;Cho, Hyung Rae;Lee, Jae Hee;Bae, Kun Yuk;Seo, Jong Jin;Moon, Hyung Nam;Im, Ho Joon
    • Clinical and Experimental Pediatrics
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    • v.53 no.2
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    • pp.210-214
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    • 2010
  • Purpose : Cisplatin is highly effective for the treatment of solid tumors in children. However, the clinical use of cisplatin is limited by its ototoxicity. The aim of this study was to evaluate the ototoxicity in children treated with cisplatin. Method : We performed a single institution retrospective analysis of pediatric oncology patients who received cisplatin therapy between January 2001 and January 2008. Thirty-seven patients with sufficient medical and audiologic data were included in this study. Results : The median age at the time of diagnosis was 10.7 (range 3.8-6.7) years. There were 16 males and 21 females. The underlying diseases were osteosarcoma (15 cases), medulloblastoma (14 cases), germ cell tumors (7 cases), and hepatoblastoma (1 case). The median individual dose was $100mg/m^2$/cycle (56-200). The median cumulative dose was $480mg/m^2$ (200-1,490). Sixteen patients (43%) received cranial radiotherapy. Of the 37 patients, 17 developed hearing loss, leading to an overall incidence of 46%. Logistic regression showed that age at treatment (P =0.04) and cumulative dose of cisplatin (P =0.005) were the significant risk factors in predicting hearing loss in children treated with cisplatin. In all the patients who had hearing loss, there was neither improvement nor aggravation during the follow-up (3-8 months). Conclusion : The cumulative dose of cisplatin (>$500mg/m^2$) and younger age at treatment (<12 years) were 2 most important risk factors for ototoxicity in patients treated with cisplatin. Serial audiometric evaluations are needed in the patients with risk factors during and after cisplatin treatment.

Serum methotrexate level is inversely related to the outcome of osteosarcoma patients (혈중 methotrexate 농도와 골육종 환자들의 치료결과의 연관성)

  • Lee, Jun Ah;Kim, Min Suk;Lee, Jin Kyung;Kim, Dong Ho;Hong, Young Joon;Song, Won Seok;Cho, Wan Hyeong;Lee, Soo-Yong;Lim, Jung Sub;Park, Kyung Duk;Jeon, Dae-Geun
    • Clinical and Experimental Pediatrics
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    • v.52 no.5
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    • pp.581-587
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    • 2009
  • Purpose : To evaluate the correlation between serum methotrexate (MTX) peak levels and clinical outcome of osteosarcoma, as well as to determine the correlation of these levels with the histologic response and event-free survival (EFS). Methods : To maintain the homogeneity of the study population, we selected 52 patients with localized extremity osteosarcoma who had received two cycles of neoadjuvant chemotherapy consisting of high-dose (HD) MTX ($12g/m^2$), cisplatin ($100mg/m^2$), and doxorubicin ($60mg/m^2$). Results : Totally, 204 courses of HD MTX were administered. The serial MTX levels ($mean{\pm}SE$) at 4 h (peak), 24 h, 48 h, and 72 h were $1292.14{\pm}12.83{\mu}m$, $9.29{\pm}3.89{\mu}m$, $1.73{\pm}1.37{\mu}m$, and $0.58{\pm}0.44{\mu}m$, respectively. The peak MTX serum level was $1292.14{\pm}12.83{\mu}m$. Neither the continuous average MTX peak level nor the dichotomized MTX peak level was related to the histologic response. However, the patients with a high 24-h MTX level ($3.4{\mu}m$) had a poor histologic response (P=0.044). An inverse relationship was observed between MTX levels and survival: the EFS was better in the patients with a mean MTX peak level of less than $1,400{\mu}m$ (P=0.002) and mean 24-h MTX level of less than $3.4{\mu}m$ (P=0.011). Conclusion : The inverse correlation between the MTX level and the outcome is an unexpected finding. Further study on the pharmacokinetics of MTX is required to substantiate our findings and elucidate the mechanism involved.

Periventricular leukomalacia induced by in utero clamping of pregnant rat aorta in fetal rats (태아 백서에서 임신 백서의 자궁 내 대동맥 결찰로 유발한 뇌실주위 백질연화증)

  • Chang, Yun Sil;Sung, Dong Kyung;Kang, Saem;Park, Soo Kyung;Jung, Yu Jin;Seo, Hyun Joo;Choi, Seo Heui;Park, Won Soon
    • Clinical and Experimental Pediatrics
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    • v.51 no.8
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    • pp.874-878
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    • 2008
  • Purpose : This study was undertaken to develop an animal model of periventricular leukomalacia (PVL) induced by in utero clamping of pregnant rat aorta in fetal rats. Methods : A timed pregnanct Sprague-Dawley rat on embryonic day 21 just prior to delivery was sedated and anesthetized, and a Harvard ventilator for small animals was applied. Following laparotomy, the maternal aorta was clamped reversibly for 40 minutes using a surgical clip. The fetal rats were then delivered by Cesarean section, resuscitated if necessary, and reared by a surrogate mother rat until postnatal day 21 to obtain the brain specimen. After systemic perfusion and fixation, $10{\mu}m$ thick serial brain sections were obtained and stained for pathologic examination and assessment of ventriculomegaly. Ventriculomegaly was assessed by the measured ventricle to total brain volume ratio. Results : Eight out of eleven fetal rats (73%) survived in the ischemia group after induction of in utero ischemia by clamping maternal rat aorta, and all ten survived in the control group. Body and brain weights measured at postnatal day 21 were significantly lower in the ischemia group compared to the control group. In pathologic findings, significant ventriculomagaly ($3.67{\pm}1.21%$ vs. $0.23{\pm}0.06%$) was observed in the ischemia group compared to the control group; although cystic lesion was not observed, mild (n=6) and moderate (n=2) rerefaction of the brain tissue was observed. Conclusion : A fetal rat model of PVL induced by in utero clamping of pregnant rat aorta was developed.

Alpha-1,3-galactosyltransferase-deficient miniature pigs produced by serial cloning using neonatal skin fibroblasts with loss of heterozygosity

  • Kim, Young June;Ahn, Kwang Sung;Kim, Minjeong;Kim, Min Ju;Ahn, Jin Seop;Ryu, Junghyun;Heo, Soon Young;Park, Sang-Min;Kang, Jee Hyun;Choi, You Jung;Shim, Hosup
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.3
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    • pp.439-445
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    • 2017
  • Objective: Production of alpha-1,3-galactosyltransferase (${\alpha}GT$)-deficient pigs is essential to overcome xenograft rejection in pig-to-human xenotransplantation. However, the production of such pigs requires a great deal of cost, time, and labor. Heterozygous ${\alpha}GT$ knockout pigs should be bred at least for two generations to ultimately obtain homozygote progenies. The present study was conducted to produce ${\alpha}GT$-deficient miniature pigs in much reduced time using mitotic recombination in neonatal ear skin fibroblasts. Methods: Miniature pig fibroblasts were transfected with ${\alpha}GT$ gene-targeting vector. Resulting gene-targeted fibroblasts were used for nuclear transfer (NT) to produce heterozygous ${\alpha}GT$ gene-targeted piglets. Fibroblasts isolated from ear skin biopsies of these piglets were cultured for 6 to 8 passages to induce loss of heterozygosity (LOH) and treated with biotin-conjugated IB4 that binds to galactose-${\alpha}$-1,3-galactose, an epitope produced by ${\alpha}GT$. Using magnetic activated cell sorting, cells with monoallelic disruption of ${\alpha}GT$ were removed. Remaining cells with LOH carrying biallelic disruption of ${\alpha}GT$ were used for the second round NT to produce homozygous ${\alpha}GT$ gene-targeted piglets. Results: Monoallelic mutation of ${\alpha}GT$ gene was confirmed by polymerase chain reaction in fibroblasts. Using these cells as nuclear donors, three heterozygous ${\alpha}GT$ gene-targeted piglets were produced by NT. Fibroblasts were collected from ear skin biopsies of these piglets, and homozygosity was induced by LOH. The second round NT using these fibroblasts resulted in production of three homozygous ${\alpha}GT$ knockout piglets. Conclusion: The present study demonstrates that the time required for the production of ${\alpha}GT$-deficient miniature pigs could be reduced significantly by postnatal skin biopsies and subsequent selection of mitotic recombinants. Such procedure may be beneficial for the production of homozygote knockout animals, especially in species, such as pigs, that require a substantial length of time for breeding.

Arthroscoic Anterior Cruciate Ligament Reconstruction with Autogenous Hamstring Gratt - Effect of the Additional Fixation after Fixation of the Graft with Intrafix - (자가 슬괵건을 이용한 관절경적 전방 십자 인대 재건술 - INTRAFIX system을 이용한 경골부 고정 후 부가적 고정의 효과 -)

  • Yoo, Jae-Doo;Roh, Kwon-Jae;Shin, Sang-Jin;Yoon, Jong-Suk;Yeo, Sung-Gu
    • Journal of the Korean Arthroscopy Society
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    • v.9 no.1
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    • pp.51-55
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    • 2005
  • Purpose: To compare the stability and clinical result after anterior cruciate ligament reconstructed knee after graft fixation using Intrafix in tibial tunnel with or without additional tibial post fixation. Materials and Methods: We analyzed 37 cases which were treated with four-strand hamstring tendon autograft during the period from May 2002 to January 2003. The grafts were fixed with Rigidfix system (Mitek Product, Johnson and Johnson, USA) in femur tunnel and Intrafix system (Mitek Product, Johnson and Johnson, USA) in tibial tunnel. After tibial fixation, additional tibial post fixation was done, which was determined by the serial case number prospectively. Patients were followed for average of fourteen months(range, thirteen to twenty-five months) At the time of final follow-up, patients were evaluated in terms of Lachman test, pivot shift test, Lysholm scores, IKDC (International Knee Documentation Committee) assessment, side-to-side KT-1000 maximum-manual arthrometer differences. Results: At last follow-up, Lysholm score was average 93.1(range: 65 to 98), IKDC assessment revealed that 26 cases had score of A, 10 cases had score of B and 1 case had score of C. The average maximum-manual KT-1000 arthrometer side ?to-side difference was 2.5 mm$(0{\sim}6mm)$. There was one case in which the Lachman test was graded as 2+ and four cases in which the Lachman test was graded as 1+ and the remaining thirty-two cases were normal by Lachman test. One case had a 2+ pivot-shift, and 2 cases had a 11 pivot-shift. The remaining 34 knees were normal on pivot -shift testing. The average maximum-manual KT-1000 arthrometer side-to-side difference was average 2.8 mm$(0{\sim}6mm)$ in Intrafix only group and average 2.2 mm$(0{\sim}4mm)$ in additional fixation group (P>0.05). Conclusion: Without additional tibial fixation, the stability of the anterior cruciate reconstructed knee with hamstring graft which was fixed with Intrafix was restored.

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