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Pathogenesis of Hantaan Virus Infection in Suckling Mice -Clinical, Virologic and Serologic Observations-

  • Kim, Gum-Ryong;Mckee, Jr, Kelly T.;Lee, Ho-Wang
    • The Journal of the Korean Society for Microbiology
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    • v.20 no.1
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    • pp.115-125
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    • 1985
  • Hemorrhagic fever with renal syndrome (HFRS) is a debilitating disease of humans caused by Hantaan virus (HV), the prototype member of a newly proposed genus of Bunyaviridae. Studies of HV pathogenesis have been limited by the absence of a well defined model for a virus-induced disease state. In an attempt to devise a model for HV pathogenesis in laboratory rodents, newborn outbred suckling ICR mice were shown to be uniformly susceptible to lethal infection with non- mouse adapted HV by intracerebral (IC), intraperitoneal (IP), intramuscular (IM), and subcutaneous (SC) inoculation routes. Clinical coures, mean time to death, and fatal outcome were age-dependent. With an inoculum of 10 $LD_{50}$, mortality was 100% in mice infected within 72h of birth, but declined to 50% by 7 days. By 2-2.5 weeks, animals developed complete resistance to clinical disease. Virus was consistently detected in serum by day 6 post-infection in IC- and IP- inoculated animals, and reached peak levels of $10^5\;PFU/ml$ by day 8 Mice infected IM and SC showed delays in onset of viremia, but achieved similar titers. Immunofluorescent antibody appeared by 17-18 days, and neutralizing antibody by 15 days, in all experimental groups. Two of 8 inbred mouse strains were identified as resistant to clinical disease : SJL/J and A/J. Manipulation of this model will allow investigation of natural rodent pathogenesis with HV, as well as offer insight into disease mechanisms and therapy of HFRS.

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Characterization and Utilization of the Clubroot Resistant Genes in Chinese Cabbage (Brassica rapa L.)

  • Hatakeyama, Katsunori
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.33-33
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    • 2015
  • Clubroot disease is the major threat to the production of Chinese cabbage (Brassica rapa L.) in Japan. Although the breeding of the clubtoot resistant (CR) cultivars is one of the most efficient ways to control this disease, the CR cultivars do not always have effects due to the breakdown of resistance. Therefore, it is necessary to develop the breeding strategy to accumulate multiple CR genes in a single cultivar effectively. We have identified two incomplete dominant CR loci, Crr1 and Crr2, which are originated from the European CR turnip Siloga. To investigate the effectiveness of marker-assisted selection (MAS) for CR breeding, the inbred line with Crr1 and Crr2 was crossed with parental lines of the existing CR $F_1$ cultivar of Chinese cabbage, followed by 5 times of MAS and backcrossing. The $F_1$ derived from a cross between the resulting parental lines improved the clubroot resistance as expected and had the same morphological characters as the original $F_1$ cultivar. We have shown that the Crr1 locus comprised two loci: Crr1a, which by itself conferred resistance to the mild isolate; and Crr1b, which had a minor effect, but was not required for Crr1a-mediated resistance. Further genetic analysis suggested that Crr1b was necessary to acquire resistance to the more virulent isolate in combination with Crr2. Molecular characterization of Crr1a encoding TIR-NB-LRR class of R protein revealed that there were at least 4 alleles in Japanese CR cultivars of Chinese cabbage. PCR analysis with Crr1a-specific markers demonstrated that the functional alleles were predicted to be present in European CR turnips, Debra and 77b besides Siloga, whereas rarely in Japanese CR cultivars, indicating that Crr1a is an useful source to improve the resistance of Chinese cabbage cultivars.

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Effect of Kami-KangHwalSan on atopic dermatitis-like skin lesions induced in NC/Nga mice by mite antigen stimulation (가미강활산(加味羌活散)이 NC/Nga mice의 아토피 발진 억제에 미치는 실험적 연구)

  • Kim, Yun-Hee;Han, Jae-Kyung;Kim, Yun-Hee
    • Journal of Haehwa Medicine
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    • v.16 no.1
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    • pp.81-91
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    • 2007
  • Objective : We wished to examine closely effect that Kami-KangHwalSan medicines used to atopy dermatitis disease patient get in atopy eruption control experimentally. Materials and Methods : Atopic dermatitis (AD) usually develops in patients with an individual or family history of allergic diseases, and is characterized by chronic relapsing inflammation seen specially in childhood, association with IgE hyperproduction and precipitation by environmental factors. However, the exact etiology of AD has been unclear. To further explore the pathogenesis and treatment of AD, a suitable animal model is required. We found that skin lesions, which were chnically and histologically very simlar to human AD, mite antigen-induced dermatitis on the face, neck, ears and dorsal skin of inbred NC/Nga mice. Results and Conclusion : Kami-KangHwalSan medicines controlled CD3+/CD69+, CD4+/CD25+, B220+/IgE+, and B220+/CD23+ revelation that an experiment that motive allergy immune reponse because an in vitro experiment stimulates splenocytes of a NC/Nga mouse same t1me by PWM, and interleukin-4, eotaxin 2, CCR3, TARC mRNA outturn that bear in splenocytes decreased remarkably by Kami-KangHwalSan medicines. Th1 cell and Th2 cell observe to be shifted by secretion amount of IL-4 and IFN-$\gamma$ by Kami-KangHwalSan medicines could know that Kami-KangHwalSan medicines can use usefully in allergy autoimmnune diease.

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Effect of Jeseupwiryeongtang-Kamibang(JWRTK) on atopic dermatitis-like skin lesions induced in NC/Nga mice by mite antigen stimulation (제습위령탕가미방(除濕胃笭湯加味方)이 NC/Nga mice의 아토피 발진 억제에 미치는 실험적 연구)

  • Na, Dong-Kyu;Kim, Yun-Hee;Han, Jae-Kyung;Kim, Yun-Hee
    • Journal of Haehwa Medicine
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    • v.16 no.1
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    • pp.93-105
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    • 2007
  • Objective : We wished to examine closely effect that Kami-JeSeubUilYeongTang medicines used to atopy dermatitis disease patient get in atopy eruption control experimentally. Materials and Methods : Atopic dermatitis (AD) usually develops in patients with an individual or family history of allergic diseases, and is characterized by chronic relapsing inflammation seen specially in childhood, association with IgE hyperproduction and precipitation by environmental factors. However, the exact etiology of AD has been unclear. To further explore the pathogenesis and treatment of AD, a suitable animal model is required. We found that skin lesions, which were clinically and histologically very similar to human AD, mite antigen-induced dermatitis on the face, neck, ears and dorsal skin of inbred NC/Nga mice. Result and Conclusion : Kami-jeseupwiryeongtang(JWRTK) medicines controlled CD3+/CD69+, CD4+/CD25+, B220+/IgE+, and B220+/CD23+ revelation that an experiment that motive allergy immune reponse because an in vitro experiment stimulates splenocytes of a NC/Nga mouse same time by PWM, and interleukin-4, eotaxin 2, CCR3, TARC mRNA outturn that bear in splenocytes decreased remarkably by Jeseupwiryeongtang-Kamibang(JWRTK) medicines. Th1 cell and Th2 cell observe to be shifted by secretion amount of IL-4 and IFN-$\gamma$ by Jeseupwiryeongtang-Kamibang(JWRTK) medicines could know that Jeseupwiryeongtang-Kamibang(JWRTK) medicines can use usefully in allergy autoimmnune diease.

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Characterization of Anthranilate Synthetase from a 5-methyltryptophan Resistant Mutant(MR1) in Maize (옥수수 5-methyltryptophan 저항성 돌연변이주(MR1)의 Anthranilate Synthetase 특성)

  • 강권규;노일섭;이효연;신동영
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.1
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    • pp.52-58
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    • 1995
  • 5-methyltryptophan(5MT) resistant mutant plants (MRl) were analyzed for characterization of anthranilate synthetase (AS) and tryptophan synthetase (TS) enzymes. The enzyme was measured in crude extracts from MR1 and control seedlings of Danggin inbred line. There was no significant difference in the level of AS between MR1 and control seedlings when grown on MS medium without 5MT. However, MR1 seedlings grown on MS medium with 25mg/L 5MT showed the level of AS twice higher than that of control seedlings. The activity of AS was inhibited to 50% in untreated plants when 4mg /L L-tryptophan was added to their extracts. Extracts from MR1 plants required about four times higher concentration of amino acid to cause equal inhibition. In the TS assay, the activity observed in MR1 seedlings was four times higher than that of control seedlings. We have also isolated and sequenced the gene which encoding the tryptophan synthetase B subunit (TSB) from maize. The gene encodes polypeptides with high homology to TSB isolated from other plants, and is expressed in all the developmental stages examined. Northern hybridization analysis indicated that the gene expression in MR1 seedlings grown on MS medium showed a higher level than in control seedlings.

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Determination of Seed Purity in Radish (Raphanus sativus L.) Using Allozyme (알로자임에 의한 무 씨의 순수성 검증)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.18 no.7
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    • pp.907-911
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    • 2008
  • Radish (Raphanus sativus L.) is one of very important crop plants in the world. It is very important to determine hybrid seed quality in the production of hybrid Brassica vegetable seeds to avoid unacceptable contamination with self-inbred (sib) seeds. The allozyme for evaluating seed purity in a commercial $F_1-hybrid$ radish cultivar is demonstrated. Three hundred sixty seeds from the male and female harvest were subsequently screened for seed purity using 27 isozyme loci. Especially, F1 hybrids of radish, Per-1 ($aa{\times}bb$), Lap-1 ($aa{\times}bb$), Est-1 ($aa{\times}bb$) were presented clear hybrid bands. Est-1 locus revealed that 15 (8.3%) seeds from the female harvest and 26 (14.4%) seeds from the male harvest were sibs. It maintains higher than average level of genetic diversity compared with their correspondent parents. Shannon's index of phenotypic diversity (I) of hybrids was the highest of all accessions (R. sativus L. cv. Daepeng, R. sativus L. cv. Backza, and their hybrids). The allozyme may lead to a better insight into the hybrid seed purity.

Optimal Condition and Interspecific Cross-Reaction of H-Y Antibody Activity (H-Y항체활성의 최적조건과 종간교차반응)

  • ;H.S.Shim;J.B.Kim;H.Y.Park;K.S.Chung
    • Korean Journal of Animal Reproduction
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    • v.10 no.2
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    • pp.168-174
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    • 1986
  • These experiments were carried out to clarify the optimalconditions and interspecific cross reaction of H-Y antibody activity. H-Y antiserum was prepared in inbred SD female rats and Balb/c female mice by repeated immunization of rat newborn testis homogemate, rat and mouse spleen cells obtained from males of same strain. The activity of H-Y antibody in antiserum was tested by ELISA and biological tests. The cross reactivity of H-Y antibody was confirmed by culturing mouse and rabbit embryos in medium containing H-Y antibody and complement obtained from rat and guinea pig, respectively. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos in medium with different pH and complement concentration. The results obtained in these experiments were summarized as follows: 1. The formation rates of H-Y antibody in rats immunized with newborn testis and spleen cell were 40.0 and 50.0% respectively, and that in mouse immunized with spleen cell was 48.4%. 2. The activity of H-Y antibody was not affected by pH in range of 6.5 to 8.0, and the same was true for the relative concentration of complement to the H-Y antibody. 3. Minimum time needed for the activity of H-Y antibody was confirmed to be 0.5 to 1 hour and 24 to 48 hours respectively for the zona free embryos and intact embryos. 4. When mouse and rabbit embryos were treated with H-Y antibody obtained from rat, 46.4 and 54.8% of embryos were retarded or destroyed. From these results it could be said that H-Y antibody had strong interspecific cross reactivity.

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Generation of Embryonic Stem Cell-derived Transgenic Mice by using Tetraploid Complementation

  • Park, Sun-Mi;Song, Sang-Jin;Choi, Ho-Jun;Uhm, Sang-Jun;Cho, Ssang-Goo;Lee, Hoon-Taek
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.121-121
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    • 2003
  • The standard protocol for the production of transgenic mouse from ES-injected embryo has to process via chimera producing and several times breeding steps, In contrast, tetraploid-ES cell complementation method allows the immediate generation of targeted murine mutants from genetically modified ES cell clones. The advantage of this advanced technique is a simple and efficient without chimeric intermediates. Recently, this method has been significantly improved through the discovery that ES cells derived from hybrid strains support the development of viable ES mice more efficiently than inbred ES cells do. Therefore, the objective of this study was to generate transgenic mice overexpressing human resistin gene by using tetrapioid-ES cell complementation method. Human resistin gene was amplified from human fetal liver cDNA library by PCR and cloned into pCR 2.1 TOPO T-vector and constructed in pCMV-Tag4C vector. Human resistin mammalian expression plasmid was transfected into D3-GL ES cells by lipofectamine 2000, and then after 8~10 days of transfection, the human resistin-expressing cells were selected with G418. In order to produce tetraploid embryos, blastomeres of diploid embryos at the two-cell stage were fused with two times of electric pulse using 60 V 30 $\mu$sec. (fusion rate : 93.5%) and cultured upto the blastocyst stage (development rate : 94.6%). The 15~20 previously G418-selected ES cells were injected into tetraploid blastocysts, and then transferred into the uterus of E2.5d pseudopregnant recipient mice. To investigate the gestation progress, two El9.5d fetus were recovered by Casarean section and one fetus was confirmed to contain human resistin gene by genomic DNA-PCR. Therefore, this finding demonstrates that tetraploid-ES mouse technology can be considered as a useful tool to produce transgenic mouse for the rapid analysis of gene function in vivo.

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Variation of Peroxidase 8 in Maize, Zea mays, L. (옥수수에 있어서 Peroxidase 8의 변이)

  • Bong Ho, Choe
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.28 no.3
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    • pp.374-378
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    • 1983
  • As one way of evaluating polymorphism in maize, variation of peroxidase 8 (Px.8) in maize plants was investigated by means of horizontal acrylomide electrophoresis. The specific part of maize plants was stele and other parts of plants were also studied for 34 different maize lines and hybrids. The results obtained indicate that Px.8 has three distinct migrating patterns on gel such as fast, slow and fast-slow. The band pattern varied with materials used. Most hybrids were showing fast-slow band, indicating that those hybrids are heterozygous at least for Px.8 allele. Variation of band pattern was obseerved within a single inbred line. The Px.8 in stele tissue and in young leaf and nodal tissue was found to be identical, indicating the possible use of those tissues as an alternative tissue for Px.8 study. It was also found that there might be some definite relationship between Px.8 and Px.3 activities in the same tissue.

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Application of Disease Resistance Markers for Developing Elite Tomato Varieties and Lines

  • Kim, Hyoun-Joung;Lee, Heung-Ryul;Hyun, Ji-Young;Won, Dong-Chan;Hong, Dong-Oh;Cho, Hwa-Jin;Lee, Kyung-Ah;Her, Nam-Han;Lee, Jang-Ha;Harn, Chee-Hark
    • Horticultural Science & Technology
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    • v.29 no.4
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    • pp.336-344
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    • 2011
  • Using the abundant available information about the tomato genome, we developed DNA markers that are linked to disease resistant loci and performed marker-assisted selection (MAS) to construct multi-disease resistant lines and varieties. Resistance markers of Ty-1, T2, and I2, which are linked to disease resistance to Tomato yellow leaf curl virus (TYLCV), Tomato mosaic virus (ToMV), and Fusarium wilt, respectively, were developed in a co-dominant fashion. DNA sequences near the resistance loci of TYLCV, ToMV, and Fusarium wilt were used for primer design. Reported candidate markers for powdery mildew-resistance were screened and the 32.5Cla marker was selected. All four markers (Ty-1, T2, I2, and 32.5Cla) were converted to cleavage amplification polymorphisms (CAPS) markers. Then, the CAPS markers were applied to 96 tomato lines to determine the phenetic relationships among the lines. This information yielded clusters of breeding lines illustrating the distribution of resistant and susceptible characters among lines. These data were utilized further in a MAS program for several generations, and a total of ten varieties and ten inbred lines were constructed. Among four traits, three were introduced to develop varieties and breeding lines through the MAS program; several cultivars possessed up to seven disease resistant traits. These resistant trait-related markers that were developed for the tomato MAS program could be used to select early stage seedlings, saving time and cost, and to construct multi-disease resistant lines and varieties.