• Title/Summary/Keyword: Inoculation technique

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Expression of Epidermal Growth Factor-like Domain 7 is Increased by Transcatheter Arterial Embolization of Liver Tumors

  • Li, Zhi;Ni, Cai-Fang;Zhou, Jin;Shen, Xiao-Chun;Yin, Yu;Du, Peng;Yang, Chao
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.1191-1196
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    • 2015
  • Background: Epidermal growth factor-like domain multiple 7 (EGFL7), recently identified as a secreted protein regulated by oxygen exposure, plays a critical role in promoting metastasis of hepatocellular carcinoma (HCC). Transcatheter arterial embolization (TAE) is widely used for treatment of HCC, resulting in hypoxia in tumors and surrounding liver tissues. Accordingly, we proposed the hypothesis that there could be a relationship between expression of EGFL7 and response to TAE. Materials and Methods: We established a rabbit VX2 liver tumor model using percutaneous puncture technique guided by computed tomography. TAE and sham embolization were performed and the results were confirmed by MRI 3 weeks after inoculation. We investigated the EGFL7 expression of the two groups at 6h and 3 days after intervention by means of immunohistochemistry and Western blotting. Results: Immunohistochemical staining demonstrated that the levels of EGFL7 protein significantly increased in the TAE-treated tumors compared with the control group at 6 hours (P=0.031) and 3 days (P=0.020) after intervention. Meanwhile, the relative EGFL7 protein detected in TAE group also up-regulated compared with the control group at 6 hours (P=0.020) and 3 days (P=0.024) after intervention. Conclusions: This study reveals an increase of EGFL7 expression in rabbit VX2 liver tumors after TAE. The role of EGFL7 in HCC, especially its biological behavior after TAE, needs further investigation.

Studies on the Comparative Migration Patterns of Ascaris suum Larvae between Primary and Re-infected Mice (돼지회충(Ascaris suum) 유충 감염력이 재감염에 미치는 영향)

  • 송종술;김재진
    • Parasites, Hosts and Diseases
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    • v.23 no.2
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    • pp.247-252
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    • 1985
  • In the present study, the effect of primary infection to reinfection with Ascaris suum larvae was experimented in mouse model. Mice were challenged with 1,000 infective stage eggs of Ascaris suum. The embryonated eggs were directly introduced into stomach of mice. Reinfection was performed at 50 days after the primary infection with same method as primary infection. Mice were sacrificed 3, 5, 7, 10, 15 and 20 days after infection in both groups respectively. Larvae collected from livers and lungs with Baermann's apparatus were enumerated and measured after sacrifice. Sera of mice were also collected at same time. The results of the experiment were as follows: With antigen prepared from coelomic fluid of adult Ascaris suum and sera collected from mice before reinfection, the production of antibody in experimental mice was confirmed by the gel-diffusion technique. In the livers of reinfected mice, the larvae were recovered up to 10 days after challenge, otherwhile in the primary infected mice, the larvae were observed up to 7 days. The maximum number of larvae were observed in the lungs of primary infected mice on 10 days after inoculation. In the lungs of reinfected mice, maximum number of larvae were recovered on 7 days after, only few larvae were recovered on 10 days after reinfection. As regards the growth of the larvae, the third stage larvae, over $500{\mu\textrm{m}}$ in length, appeared in livers at 5 days after reinfection, but it couldn't be found on 7 days and 10 days after challenge. The third stage larvae continuously developed were observed in lungs of mice from 5 days after reinfection. In conclusion, it was found that development of larvae in livers of immune mice were probably repressed by the immune mechanisms being rises in livers and defence mechanism is also acting by interfering with the process of larval penetration into the lung from the liver.

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Identification of Fusarium Species Associated with Corn Ear Rot (옥수수 이삭썩음병에 관여하는 Fusarium속균의 동정)

  • Choi, Hyo-Won;Kim, Jung-Mi;Kim, Jin-Hee;Hong, Sung-kee;Kim, Wan-Gyu;Chun, Se-Chul
    • The Korean Journal of Mycology
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    • v.37 no.2
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    • pp.121-129
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    • 2009
  • In 2007, a total of 77 isolates of Fusarium spp. were obtained from ear rot symptoms of corns collected from 5 locations in Gangwon Province, Korea. The fungal isolates were identified based on their morphological features. Out of the isolates, fifteen isolates were identified as Fusarium verticillioides which formed microconidia in long chains on monophialides. Four isolates were identified as F. subglutinans which formed microconida only on false heads. Six isolates were identified as F. graminearum which produced red pigment in PDA culture. Besides these Fusarium species, F. napiform, F. nygamai, and F. oxysporum were identified from the rest isolates. To assess for genetic diversity of the isolates, a random amplified polymorphic DNA(RAPD) technique was carried out using URP primers. The results from the RAPD analysis showed that the isolates from corn were divided into 6 groups. These RAPD groups of the Fusarium species corresponded to morphological characters of the Fusarium species. The phylogenetic analysis of most isolates by DNA sequencing of EF-1$\alpha$ gene corresponded to morphological characters of the Fusarium species. The results of pathogenicity tests by two inoculation methods revealed that F. verticillioides, F. graminearum and F. subglutinans are strongly pathogenic to corn stalks.

Recolonization of Transfected Blastodermal Cells in Developing Embryos after Transferring into UV-irradiated Fertilized Hen′s Egg (UV-조사 수정란 내로 이식한 유전자 변화 배반엽 세포의 재구성)

  • Lee, K.S.;Lee, H.;Kim, K.D.;Park, S.S.;Lee, S.H.
    • Korean Journal of Poultry Science
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    • v.27 no.2
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    • pp.155-161
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    • 2000
  • Unfortunately, there is no technique which is stable and repetitive to produce transgenic chicken, although various ways of gene transfer including PGC-and embryonic cell-mediated gene transfer, DNA microinjection, virus inoculation and sperm cells have been employed. The aims of this study were 세 develop and establish such a stable, repetitive and efficient way of gene transfer giving a faithful gene expression during development after the reconstruction of embryo in an UV-irradiated egg. A dual reporter plasmid (pJJ9), a fusion gene containing lacZ and GFP driven by a CMV promoter was used to exploit either merits of both reporting markers. lacZ with strong signal or GFP with vital marking. Electroporated embryonic blastodermal cells (EBCs) in the presence of the pJJ9 DNA faithfully showed 377 bp PCR product and lacZ or GFP expressions in the identical cells in vitro of in vivo. Furthermore, analyses of expression pattern of the foreign DNA demonstrated that microinjected EBCs cells into the UV-irradiated recipient egg should participate in normal developmental process, for example, proliferation and differentiation into various tissues. Thirty percentages of the manipulated eggs showed lacZ expression in their tissues. These results together with the specific procedures used in this study should facilitate avian transgenesis.

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Isolation of Sulfur Oxidizing Bacteria from Different Ecological Niches (다양한 생태계에서 분리한 황 산화 세균)

  • Anandham, Rangasamy;Sridar, Ragupathy;Nalayini, Periyakaruppan;Madhaiyan, Munusamy;Gandhi, Pandiyan Indira;Choi, Kwan-Ho;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.4
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    • pp.180-187
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    • 2005
  • Nine chemolithoautotrophic and 12 chemolithoheterotrophic sulfur oxidizing bacteria were isolated using enrichment technique in modified Starkey's medium. All isolates reduced pH of the growth medium through oxidation of elementai sulfur to sulfuric acid. Isolates utilized the thiosulfate as energy source except LCH. None of the isolates grew anaerobically and utilization of glucose was found only in chemolithoheterotrophic isolates SGA6 and JIG. In vitro sulfate production from elemental sulfur was found maximum for chemoiithoautotroph LCH ($43.2mg\;100\;mL^{-1}$) and least for chemolithoheterotroph JIG ($10.04mg\;100\;mL^{-1}$). The above tests suggested that all isolates belong to the member of Thiobacillus. For field inoculation of Thiobacillus, clay based pellet formulation was developed with the cell load of $2.5{\times}10^7cfu\;g^{-1}$ of pellet. It is easy to handle by the farmers and more likely to lead to successful farming.

Classical swine fever disease in Cheolwon

  • Park Yang-Soon;Shin Myung-Kyun;Chong Dong-Soo;Cheong Ki-Soo;Park Young-Nam;Choi Jee-Hee
    • Korean Journal of Veterinary Service
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    • v.27 no.4
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    • pp.345-354
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    • 2004
  • Two cases of classical swine fever (CSF) disease have broken out in Cheolwon (7 April, 2002). The suspected pig herds were huddled together because of high fever (over $40^{\circ}C$) and showed remarkable decrease of the leukocytes. The staggering gait related to posterior weakness, constipation and lethargy, hyperemia, hemorrhagic lesions (on the skin, muzzle, ears, limbs, tail and inner part of legs) and conjunctivitis with dirty streaks below the eyes were observed. The inflammation in the lung, infarction in the spleen, swelling and hemorrhage in lymph nodes, kidney, intestine, heart and cheese like purulent inflammation of the tonsil were observed. The ulcers of the colon were also detected. Several clinical and laboratory techniques including blood test, histo-pathological examinations, indirect fluorescent antibody (IFA) test and RT-PCR test were applied to diagnose the disease. Inoculation test on PK-15 cell was also performed. The necrosis of the lymphatic cells and infiltration of the vessel circumferential cells in the brain and lymph organs were commonly viewed. The proliferation of the glia cell (gliosis) in the lymph was particular. Cytopathogenic effect (CPE) and specific fluorescent-bright-green areas (with IFA) appeared in PK-15 cells inoculated with suspected blood plasma. The IFA test on the epithelial and mucous membrane cells of tonsil was positive. RT-PCR technique required more working hours and labor than other techniques in this examination but it was useful because of the sensitivity to the CSF viral gene.

Identification of New Isolates of Phytophthora sojae and Selection of Resistant Soybean Genotypes

  • Su Vin Heo;Hye Rang Park;Yun Woo Jang;Jihee Park;Beom Kyu Kang;Jeong Hyun Seo;Jun Hoi Kim;Ji Yoon Lee;Man Soo Choi;Jee Yeon Ko;Choon Song Kim;Sungwoo Lee;Tae-Hwan Jun
    • The Plant Pathology Journal
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    • v.40 no.3
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    • pp.329-335
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    • 2024
  • Phytophthora root and stem rot (PRR), caused by Phytophthora sojae, can occur at any growth stage under poorly drained and humid conditions. The expansion of soybean cultivation in South Korean paddy fields has increased the frequency of PRR outbreaks. This study aimed to identify four P. sojae isolates newly collected from domestic fields and evaluate race-specific resistance using the hypocotyl inoculation technique. The four isolates exhibited various pathotypes, with GJ3053 exhibiting the highest virulence complexity. Two isolates, GJ3053 and AD3617, were screened from 205 soybeans, and 182 and 190 genotypes (88.8 and 92.7%, respectively) were susceptible to each isolate. Among these accessions, five genotypes resistant to both isolates were selected. These promising genotypes are candidates for the development of resistant soybean cultivars that can effectively control PRR through gene stacking.

Inflammation Scan Using $^{99m}Tc-HMPAO$ Labelled Leukocytes ($^{99m}Tc-HMPAO$를 이용한 자가백혈구표지 및 그를 이용한 염증병소의 스캔)

  • Yang, Woo-Jin;Chung, Soo-Kyo;Shinn, Kyung-Sub;Bahk, Yong-Whee;Kim, Hoon-Kyo
    • The Korean Journal of Nuclear Medicine
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    • v.23 no.2
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    • pp.219-223
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    • 1989
  • Inflammation scan using radiolabelled leukocytes has high sensitivity and specificity. Several methods for labelling leukocytes have been evaluated using P-32 diisopropyl fluorophosphate (DFP-32), H-3 thymidine, Cr-51 chromate, Ga-67 citrate and Tc-99m-sulfur colloid. In-111-oxine has proved so far to be the most reliable agent for labelling leukocytes. In-111-oxine is, however, expensive, not easily available when needed, and its radiation dose to leukocytes is relatively high. Moreover, resolution of the resultant image is relatively poor. Tc-99m is still the agent of choice because of, as compared with the indium, its favorable physical characteristics, lower cost and availability. Now the technique for labelling the leukocytes with technetium is successfully obtained using the lipophilic HAPAO with higher efficiency for granulocytes than for other cells. With this technique it is possible to label leukocytes in plasma to improve the viability of the leukocytes. Inflammation scan using Tc-99m-HMPAO has been evaluated in several laboratories, and difference in methods for separation and labelling accounts for difference in efficiency, viability and biodistribution of the labelled leukocytes. We performed inflammation scan using leukocytes labelled with Tc-99m-HMPAO in three dogs 24 hours after inoculation of live E. Coli and A. Aureus in their right abdominal wall. We separated mixed leukocytes by simple sedimentation using 6% hetastarch (HES) and labelled the leukocytes with Tc-99m-HMPAO in 20% cell free plama diluted with phosphate buffer solution(Fig. 1). Uptake was high in the liver and spleen but is was minimal in the lungs on whole body scan. Kidneys and intestine showed minimal activity although it was high in the urinary bladder(Fig. 2). Uptake of labelled leukocytes in the inflammation site was do(mite on 2 hour-postinjection scan and abscess was clearly delineated on 24 hour-delayed scan with high target-to-nontarget ratio(Fig. 3, 4). Inflammation scan using mixed leukocytes labelled with Tc-99m-HMPAO is very sensitive and specific in early detection of inflammation.

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Control of Tomato Wilt Disease by Amending pH of Nutrient Solution in Hydroponic System (토마토 수경재배에서 배양액의 pH 조절에 의한 풋마름병 방제)

  • Lee Jung-Sup;Choi Ji-Ho;Seo Sang-Tae;Han Kyoung-Suk;Park Jong-Han;Jang Han-Ik
    • Research in Plant Disease
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    • v.11 no.2
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    • pp.193-197
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    • 2005
  • The effect of pH on the survival of R. solanacearum and its transmission via roots of tomato in hydroponic culture were studied in laboratory and greenhouse. In laboratory experiment, R. solanacearum could not survive for 24h in nutrient solution with pH of $4{\cdot}0;or\;4{\cdot}5$, while 1, 14, 51 and $62\%$ of inoculum survived at pH $5{\cdot}0,\;5{\cdot}6\;and\;6{\cdot}5$, respectively. When tomato plants were inoculated with R. solanacearum through wounds on the stems, the bacteria moved downward from the inoculation site to the roots and infectious bacteria were released from the roots into the nutrient solution. Of two pH regimes tested in greenhouse nutrient-film technique(NFT) culture, the R. solanacearum population was significantly lower in pH 5.0 than in pH 6.5 in most sampling data. In treatments in which R. solanacearum was introduced by transplanting two root-inoculated plants, significantly move plants developed wilt at pH $6{\cdot}5$(34 out of 48 plants) than at pH 5.0(11 out of 48 plants). In addition, when the bacterium was introduced by transplanting two stem-inoculated plants at pH $6{\cdot}5$, seven out of 24 plants developed wilt.

Studies on Garlic Mosaic Virus -lts isolation, symptom expression in test plants, physical properties, purification, serology and electron microscopy- (마늘 모자이크 바이러스에 관한 연구 -마늘 모자이크 바이러스의 분리, 검정식물상의 반응, 물리적성질, 순화, 혈청반응 및 전자현미경적관찰-)

  • La Yong-Joon
    • Korean journal of applied entomology
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    • v.12 no.3
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    • pp.93-107
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    • 1973
  • Garlic (Allium sativum L.) is an important vegetable crop for the Korean people and has long been cultivated extensively in Korea. More recently it has gained importance as a source of certain pharmaceuticals. This additional use has also contributed to the increasing demand for Korean garlic. Garlic has been propagated vegetatively for a long time without control measures against virus diseases. As a result it is presumed that most of the garlic varieties in Korea may have degenerated. The production of virus-free plants offers the most feasible way to control the virus diseases of garlic. However, little is known about garlic viruses both domestically and in foreign countries. More basic information regarding garlic viruses is needed before a sound approach to the control of these diseases can be developed. Currently garlic mosaic disease is most prevalent in plantings throughout Korea and is considered to be the most important disease of garlic in Korea. Because of this importance, studies were initiated to isolate and characterize the garlic mosaic virus. Symptom expression in test plants, physical properties, purification, serological reaction and morphological characteristics of the garlic mosaic virus were determined. Results of these studies are summarized as follows. 1. Surveys made throughout the important garlic growing areas in Korea during 1970-1972 revealed that most of the garlic plants were heavily infected with mosaic disease. 2. A strain of garlic mosaic virus was obtained from infected garlic leaves and transmitted mechanically to Chenopodium amaranticolor by single lesion isolation technique. 3. The symptom expression of this garlic mosaic virus isolate was examined on 26 species of test plants. Among these, Chenopodium amaranticolor, C. quince, C. album and C. koreanse expressed chlorotic local lesions on inoculated leaves 11-12 days after mechanical inoculation with infective sap. The remaining 22 species showed no symptoms and no virus was recovered from them whet back-inoculated to C. amaranticolor. 4. Among the four species of Chtnopodium mentioned above, C. amaranticolor and C. quinoa appear to be the most suitable local lesion test plants for garlic mosaic virus. 5. Cloves and top·sets originating from mosaic infected garlic plants were $100\%$ infected with the same virus. Consequently the garlic mosaic virus is successively transmitted through infected cloves and top-sets. 6. Garlic mosaic virus was mechanically transmitted to C, amaranticolor when inoculations were made with infective sap of cloves and top-sets. 7. Physical properties of the garlic mosaic virus as determined by inoculation onto C. amaranticolor were as follows. Thermal inactivation point: $65-70^{\circ}C$, Dilution end poiut: $10^-2-10^-3$, Aging in vitro: 2 days. 8. Electron microscopic examination of the garlic mosaic virus revealed long rod shaped particles measuring 1200-1250mu. 9. Garlic mosaic virus was purified from leaf materials of C. amaranticolor by using two cycles of differential centrifugation followed by Sephadex gel filtration. 10. Garlic mosaic virus was successfully detected from infected garlic cloves and top-sets by a serological microprecipitin test. 11 Serological tests of 150 garlic cloves and 30 top-sets collected randomly from seperated plants throughout five different garlic growing regions in Korea revealed $100\%$ infection with garlic mosaic virus. Accordingly it is concluded that most of the garlic cloves and top-sets now being used for propagation in Korea are carriers of the garlic mosaic virus. 12. Serological studies revealed that the garlic mosaic virus is not related with potato viruses X, Y, S and M. 13. Because of the difficulty in securing mosaic virus-free garlic plants, direct inoculation with isolated virus to the garlic plants was not accomplished. Results of the present study, however, indicate that the virus isolate used here is the causal virus of the garlic mosaic disease in Korea.

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