• Title/Summary/Keyword: In vivo

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Molecular Breeding of Tobacco Plants Resistant to TMV and PVY (분자생물학적 TMV 및 PVY 저항성 연초 육종)

  • E.K. Pank;Kim, Y.H.;Kim, S.S.;Park, S.W.;Lee, C.H.;K.H.Paik
    • Proceedings of the Korean Society of Tobacco Science Conference
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    • 1997.10a
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    • pp.134-152
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    • 1997
  • Plant viruses of tobacco including tobacco mosaic virus (TMV) and potato virus Y (PVY) cause severe economic losses in leaf-tobacco production. Cultural practices do not provide sufficient control against the viruses. Use of valuable resistant cultivars is most recommendable for the control of the viruses. However, conventional breeding programs are not always proper for the development of virus-resistant plants mostly owing to the frequent lack of genetic sources and introduction of their unwanted properties. Therefore, we tried to develop virus-resistant tobacco plants by transforming commercial tobacco cultivars, NC 82 and Burley 21, with coat protein (CP) or replicase (Nlb) genes of TMV and PVY necrosis strain (PVY-VN) with or without untranslated region (UTR) and with or without mutation. Each cDNA was cloned and inserted in plant expression vectors with 1 or 2 CaMV 35S promotors, and introduced into tobacco leaf tissues by Agrobacterium tumefaciens LBA 4404. Plants were regenerated in kanamycin-containing MS media. Regenerated plants were tested for resistance to TMV and PVY In these studies, we could obtain a TMV-resistant transgenic line transformed with TMV CP and 6 genetic lines with PVY-VN cDNAs out of 8 CP and replicase genes. In this presentation, resistance rates, verification of gene introduction in resistant plants, stability of resistance through generations, characteristics of viral multiplication and translocation in resistant plants, and resistance responses relative to inoculum potential and to various PVY strains will be shown. Yield and quality of leaf tobacco of a promising resistant tobacco line will be presented.

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The Morphological Study on Inflammation of Murine Knee Joint by Lipopolysaccharide - Based on the Morphological Changes of Synovial Membrane and Fibrous Membrane - (Lipopolysaccharide로 유발된 생쥐 무릎관절낭 염증에 관한 형태학적 연구 - 윤활관절막과 섬유관절막의 변화를 중심으로 -)

  • Kim, Jin-Tack;Ahn, Sang-Hyun;Choi, Nan-Hee;Chung, Jae-Man;Park, In-Sick;Gang, Yun-Ho;Kim, Ho-Hyun;Lee, Hai-Poong
    • The Journal of Dong Guk Oriental Medicine
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    • v.7 no.2
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    • pp.107-120
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    • 1999
  • Synovial joint of BALB/C mice were injeced with Lipopolysaccharide(LPS) were observed to investigate the morphological changes of synovial capsule caused by rheumatoid arthritis(RA). The RA on female Balb/c mice were induced by LPS injection, as dose of $300{\mu}{\ell}/kg$, into synovial cavity of knee joint. And then these specimen were fixed in 10% neutral buffered formalin and were decalcificated in EDTA solution for 4 weeks. The hyperplasia of synovium were appeared in synovial membrane. The filopodia of phagocytic like synoviocyte(type I synoviocyte) projected into synovial cavity and the number of fibroblast like synoviocyte(type II synoviocyte) with well-developed endoplasmic reticulum were increased in synovium. In fibrous membrane, the fibrosis induced by synthesis of collagen fiber were enlarged to all fibrous membrane, and the number of fibroblast were increased. A great number of inflammation component cell as lymphocyte and neutrophil leukocyte were infiltrated around capillary and the degranulate typed mast cell were increased. As results indicated that the hyperplasia of synovium induced by LPS, subsequently to cause the fibrosis, infiltration of imflammation component cell, and increase of degranulated type mast cell as same as symptoms of RA.

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Immuno-Enhancing Effect of Enzymatic Extract of Sargassum coreanum Using Crude Enzyme from Shewanella oneidensis PKA 1008 (Shewanella oneidensis PKA 1008 유래 조효소 처리에 의한 큰잎모자반(Sargassum coreanum) 추출 분해물의 면역증진 효과)

  • Park, Sun-Hee;Kim, Min-Ji;Kim, Go-Eun;Park, So-Yeong;Kim, Koth-Bong-Woo-Ri;Kim, Yeon-Ji;Cho, Young-Je;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.8
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    • pp.919-928
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    • 2017
  • The immuno-enhancing effects of alginate oligosaccharides from Sargassum coreanum were investigated. The alginate oligosaccharides were produced by an alginate-degrading enzyme from S. oneidensis PKA 1008. The degraded alginate oligosaccharides were visualized by thin-layer chromatography developed using a solvent system of 1-butanol/methanol/water, 4:1:2 (v/v/v). Alginate was degraded into dimmers at 60 h. As a result, the levels of Th1 cytokine [interferon $(IFN)-{\gamma}$ and interleukin (IL)-2] and Th2 cytokine (IL-6 and IL-10) increased with increasing incubation time compared to the control in vitro. Enzymatic extract treatment promoted proliferation of splenocytes at concentrations of 100 and 200 mg/kg at 24 h in vivo. Secretion of $IFN-{\gamma}$ and IL-2 significantly increased in a dose-dependent manner at 24 h as well as induced higher production of IgG2a in serum. Natural killer cell activity was measured and tended to increase. In addition, complete blood cell counts increased in a dose-dependent manner. These results indicate that alginate oligosaccharides produced by crude enzyme from S. oneidensis PKA 1008 may have significant immune activities.

DEVELOPMENT OF HIGH SENSITIVE MODEL OF CARIES ACTIVITY TEST FOR EARLY DIAGNOSIS OF DENTAL CARIES (치아우식증의 조기진단을 위한 고감도 우식활성검사 모형개발)

  • Lee, Sang-Ho;Lee, Chang-Seop
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.1
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    • pp.169-179
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    • 2000
  • The purpose of this study is to develop the system which convert the optical difference of teeth texture between intact enamel and incipient caries lesion into shade difference by laser fluorescence and to develop new and simple caries activity test using laser fluorescence. The experimental design of this study consists of three parts. In first part, a new method for the in vitro assessment of changes in initial enamel caries lesion of Bovine teeth using laser fluorescence is tested. In second part, in vivo assessment undertaken. Number of teeth which showed incipient carious lesion on buccal surface examined by laser fluorescence was compared with the caries activity test of $Cariescreen^{(R)}$ test and other oral environmental test of dDfFtT. In third part, new caries activity test measured by laser fluorescence was developed on the basis of above results and evaluated the sensitivity, specificity, and diagnostic power. Optical density measured by laser fluorescence was increased as increasing the depth of incipient carious lesion and showed high correlation$(\gamma=0.7015)$ with lesion depth. Optical density showed direct proportion to lesion depth. Linear equation was obtained between the optical density and the lesion depth by regression analysis. The result of caries activity test with laser fluorescence showed high correlation with those of $Cariescreen^{(R)}$ test and dDfFtT examination. Caries activity test with laser fluorescence showed 48% of sensitivity, 52% of specificity, and 45% of diagnostic power on the basis of dDfFtT examination, and also showed 48% of sensitivity, 51% of specificity, and 36% of diagnostic power on the basis of $Cariescreen^{(R)}$ test. In regard above result, caries activity test with laser fluorescence considered to be reliable for caries activity test compared with other oral environmental test. and it was also considered to be practical because it would be simple, inexpensive, and time saving method.

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Anti-Obesity Effects of Jeju Hallabong Tangor (Citrus kiyomi${\times}$ponkan) Peel Extracts in 3T3-L1 Adipocytes (제주산 한라봉 과피 추출물의 지방세포에서의 항비만 효과)

  • Lim, Heejin;Seo, Jieun;Chang, Yun-Hee;Han, Bok-Kyung;Jeong, Jung-Ky;Park, Su-Beom;Choi, Hyuk-Joon;Hwang, Jinah
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.11
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    • pp.1688-1694
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    • 2014
  • Jeju Hallabong Tangor (Citrus kiyomi${\times}$ponkan) is a Citrus species with a variety of physiological properties such as anti-oxidant, anti-inflammation, anti-cancer, and anti-obesity. We investigated the anti-obesity effects of Hallabong Tangor peel extracts before (HLB) and after (HLB-C) bioconversion with cytolase based on modulation of adipocyte differentiation and lipid metabolism in 3T3-L1 adipocytes. Treatment with cytolase decreased flavanone rutinoside forms (narirutin and hesperidin) and increased flavanone aglycone forms (naringenin and hesperetin). During adipocyte differentiation, 3T3-L1 cells were treated with 0.5 mg/mL of Sinetrol (a positive control), HLB or HLB-C. Adipocyte differentiation was inhibited in both citrus groups, but not in control and Sinetriol groups. HLB and HLB-C tended to reduce insulin-induced mRNA levels of CCAAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$) and sterol regulatory element-binding protein 1c (SREBP1c). Compared to the control and Sinetrol groups, HLB and HLB-C markedly suppressed insulin-induced protein expression of $C/EBP{\alpha}$ and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$). The HLB and Sinetrol groups, but not HLB-C group, significantly increased adipolytic activity with higher release of free glycerol compared to the control group in differentiated 3T3-L1 adipocytes. These results suggest that bio-conversion of Hallabong Tangor peel extracts with cytolase increases aglycone flavonoids. Irrespective of bioconversion, both Hallabong Tangor peel extracts exert anti-obesity effects that may contribute to prevention of obesity through inhibition of adipocyte differentiation or induction of adipolytic activity.

Carboxyhemoglobin Dissociation at the Various Partial Pressures of Oxygen -Comparison between Adult and Fetal Bloods- (산소분압(酸素分壓)에 따른 Carboxyhemoglobin의 해리양상(解離樣相) -성인혈(成人血)과 태아혈(胎兒血)의 비교(比較)-)

  • Park, Byung-Joo
    • Journal of Preventive Medicine and Public Health
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    • v.15 no.1
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    • pp.145-151
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    • 1982
  • Breslau's report on the two stillbirths induced by illuminating gas poisoning made many investigators explore the hazards. of carbon monoxide(CO) poisoning to pregnancy. The pregnant woman, her fetus, and the newborn infant have been identified to be particularly vulnerable to CO even in low concentration. Several factors, such as placental barrier, membrane resistance of maternal and fetal red blood cells etc., were considered to be related to the delayed elimination of CO from fetus. Slower elimination of CO from fetus than from mother was confirmed in several in vivo studies. But there are few studies which have confirmed the difference of carboxyhemoglobin (HbCO) dissociation in adult and fetal bloods. Author investigated the effects of hemoglobin itself on the elimination of CO from mother and fetus. By observing the difference of CO dissociation from adult and fetal hemoglobin at the various partial pressures of oxygen, the author tries to suggest the base of the proper treatment measure for the CO poisoning of pregnant woman and newborn infant. The results were as follows: 1. The total hemoglobin amounts of adults and fetal bloods were $16.1{\pm}0.50gm%\;and\;15.7{\pm}0.32gm%$, respectively. The fetal hemoglobin proportions in adult and fetal bloods were $1.2{\pm}0.15%\;and\;72.7{\pm}3.01%$, respectively. 2. Adult and fetal bloods saturated by CO to 100% HbCO were exposed to ambient air$(21%\;O_2),\;100%\;O_2\;and\;3\;ATAO_2$. After 30 minutes exposure, the HbCO saturations of adult blood were 96.7%, 70.9%, and 52.8%, respectively, and those of fetal blood were 98.5%, 76.1%, and 62.2%, respectively. HbCO dissociation was proportional to the partial pressure of oxygen and the most marked dissociation was shown under 3 ATA $O_2$, HbCO dissociation of fetal blood was slower than that of adult blood in all conditions. According to the above results, it is possible that CO poisoning make more serious damage to the fetus and newborn infant than to the adult due to the delayed dissociation of HbCO. Thus in the treatment of CO poisoning of pregnant woman and newborn infant, hyperbaric oxygen therapy seems to be the most eflective treatment measure, but the duration of hyperbaric oxygenation should be lengthened accordingly.

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Anti-Thrombogenic and Anti-Inflammatory Effects of Solvent Fractions from Leaves of Zanthoxylum Schinifolium (Sancho Namu) in Rats Fed High Fat Diet (고지방식이 흰쥐에서 산초나무 Butanol 및 Methylene Chloride 분획의 항혈전 및 항염증 작용)

  • Jang, Hyun-Seo;Rhee, Soon-Jae;Woo, Mi-Hee;Cho, Sung-Hee
    • Journal of Nutrition and Health
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    • v.40 no.7
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    • pp.606-615
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    • 2007
  • This study was performed to investigate anti-thrombogenic, anti-inflammatory effects of n-BuOH (B) and $CH_2Cl_2$ (MC) fractions extracted from Sancho (Zanthoxylum. schinifolium) leaves in rats fed high fat diets. The experimental animal groups were consisted of eight including one 5% fat (N) and one 20% fat (H) without the test materials in diets and six H groups of feeding three levels (50, 100 and 150 mg/day) of the B and the MC fractions from Z. schinifolium, respectively. Plasma activated partial thromboplastin times and thrombin times of H group were decreased compared to the N group, but they were increased by feeding the MC fraction of 50 mg and over. Polymorphonuclear leukocyte 5#-lipo-xygenase activities and leukotriene $B_4$ contents of the H group were significantly increased compared to the N group, but they were decreased in the 100 mg and 150 mg of B fraction or the 150 mg of MC fraction fed groups. Liver cytochrome $P_{450}$, $O_2^-$, $H_2O_2$ and GSSG contents were increased by the high fat diet but decreased by feeding the B fraction or the MC fraction, while GSH content and glutathione S-transferase activity lowered by high fat diet were increased by feeding the two solvent fractions. The effects of the solvent fractions were evident at the level of 100 mg/day and over. The present results confirmed that two solvent fractions from the leaves of Z, schinifolium have enhancing effects on anti-thrombosis and anti-inflammation partly by antioxidant action and partly by direct modulation of the respective processeds. In conclusion, the n-BuOH and $CH_2Cl_2$ fractions from leaves of Z, schinifolium can be utilized as the proper ingredients of functional foods for preventing chronic degenerative disease.

In vivo and in vitro Confirmation of Dose Homogeneity in Total Body Irradiation with Thermoluminescent Dosimeter (인체 및 인형 팬톰에서 전신방사선조사시 열형광선량계(TLD)를 이용한 선량분포 균일성 확인)

  • Chie Eui Kyu;Park Suk Won;Kang Wee-Saing;Kim Il Han
    • Radiation Oncology Journal
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    • v.17 no.4
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    • pp.321-328
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    • 1999
  • Purpose : Total body irradiation (TBI) or whole body irradiation is used to acquire immune suppression, to treat malignant lymphoma and leukemia, and as an conditioning regimen for bone marrow transplantation. For these purposes, many methods were developed to obtain homogenous dose distribution. The objective of this study was to analyze and confirm the accuracy and the homogeneity of the treatment setup, the parallel opposed lateral technique, currently used in Seoul National University Hospital. Materials and Metheods : Surface dose data, measured with a thermoluminescent dosimeter, of 8 patients among 10 patients, who were given total body irradiation with the parallel opposed lateral technique between September 1996 to August 1998, at Seoul National University Hospital were analyzed. Surface doses were measured at the head, neck, axilla, thigh, and ankle level. Surface and midline doses were measured with similar set-up and technique in the Humanoid phantom. Results : Measured surface doses relative to prescribed dose for the head, neck, axilla, thight, and ankle leve were $91.3{\pm}7.8,{\;}98.3{\pm}7.5,{\;}95.1{\pm}6.3,{\;}98.3{\pm}5.5$, and $95.3{\pm} 6.3\%$, respectively. The midline doses of the head, neck, axilla, thigh, and ankle level estimated from the surface-to-midline ratios in the Humanoid Phantom were $103.4{\pm}9.0,{\;}107.8{\pm}10.5,{\;}91.1{\pm}6.1,{\pm} 93.8{\pm}4.5,{\;}and{\;}104.5{\pm}9.3\%$, respectively. Measured surface doses and estimated midline doses ranged from $-8.9\%$ to $+7.8\%$. Midline doses at the neck and the axilia level deviated more than $5\%$ from the prescribed doses. The difference of the estimated midline doses at the neck and the axilla level and the actual doses were attributed to the thickness differences between the Humanoid phantom and the patients. Conclusion Distribution of the midline doses as well as the suface doses were measured to be within $-8.7\~{\pm}7.8\%$ range. Actual dose distribution in the patient is expected to be better than the measured dose range mainly attributed to thickness difference between the patient and the Humanoid phantom.

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Antithrombotic Activities of Cheongkookjang and Cheongkookjang Fermented with Green Tea or Mugwort (청국장 및 녹차, 쑥이 첨가된 청국장의 항혈전 활성)

  • Lee, Kyung-Ae;Jang, Jeong-Oak;Yoon, Hye-Kyung;Kim, Moo-Sung
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.298-303
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    • 2007
  • Antithrombotic activities of water extract of cheongkookjang and cheongkookjang fermented with green tea or mugwort were evaluated on some antithrombosis related activities in vitro and thrombotic death inhibition in vivo. Cheongkookjang made of white soybean (Glycine max) or black small soybean (Rhynchosia nulubilis) showed potent antioxidative activities. Addition of green tea or mugwort during cheongkookjang fermentation increased the antioxidative activity, cheongkookjang with green tea showed more drastic increase compared with cheongkookjang with mugwort. Nitrite scavenging effects of the cheongkookjang extracts were prominent but the addition of green tea or mugwort seldom increased the scavenging effects. All the cheongkookjang extracts showed strong inhibitory activities on platelet aggregation. The inhibitory activities of cheongkookjang were increased considerably by addition of green tea or mugwort even with low concentration. Plasmin unit as fibrinolytic activity was not affected considerably by addition of green tea. Addition of mugwort decreased the activity transiently at low concentration ($0.3{\sim}1.0%$) but increased again slowly at higher concentration ($1{\sim}3%$). In vitro thrombotic death inhibition test, the antithrombotic activity of cheongkookjang made of black small bean with green tea was higher by about 1.5 times compared to that without green tea. As results, cheongkookjang might inhibit antithrombosis not only by fibrinolytic action but also by inhibition of platelet aggregation and antioxidative action. The addition of functional materials such as green tea or mugwort could increase the antithrombotic function, even at low concentration.

Apoptotic Effect of Co-Treatment with Valproic Acid and HS-1200 on Human Osteosarcoma Cells (Valproic acid와 HS-1200의 병용처리가 사람골육종세포에 미치는 세포자멸사 효과에 대한 연구)

  • Kim, Duck-Han;Lee, Kee-Hyun;Kim, In-Ryoung;Kwak, Hyun-Ho;Park, Bong-Soo;Jeong, Sung-Hee;Ko, Myung-Yun;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.35 no.3
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    • pp.165-175
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    • 2010
  • Valproic acid (VPA) is a well-known anticonvulsive agent and has been used in the treatment of epilepsy for almost 30 years. VPA emerged in 1997 as an antineoplastic agent as well, when findings indicated the substance inhibited proliferation and induced differentiation of primitive neuroectocdermal tumor cells in vivo (Cinatl et al., 1997). Antitmor activity of VPA is associated with its targeting histone deacetylases. Bile acids and their synthetic derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. It has been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis-inducing activity on various cancer cells in vitro. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with the histone deacetylases inhibitor, VPA and a CDCA derivative, HS-1200 on human osteosarcoma (HOS) cells. Cell viability was evaluated by trypan-blue exclusion. Induction and augmentation of apoptosis were confirmed by Hoechst staining, flow cytometry (DNA hypoploidy and MMP change), Westen blot analysis and immunofluorescent staining. In this study, HOS cells co-treated with VPA and HS-1200 showed several lines of apoptotic manifestation such as nuclear condensations, the reduction of MMP, the decrease of DNA content, the release of cytochrome c into cytosol, the translocation of AIF onto nuclei, and activation of caspase-7, caspase-3 and PARP whereas each single treated HOS cells did not. Although the single treatment of 1 mM VPA or $25\;{\mu}M$ HS-1200 for 48 h did not induce apoptosis, the co-treatment of them induced prominently apoptosis. Therefore our data provide the possibility that combination therapy of VPA and HS-1200 could be considered as a novel therapeutic strategy for human osteosarcoma.