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Detection and Genotyping of Viruses Detected in Children with Benign Afebrile Seizures Associated with Acute Gastroenteritis (급성 위장관염에 병발하는 양성 무열성 경련 소아 환자의 대변에서 검출된 바이러스 및 유전자형 분석 연구)

  • Yang, Hye-Ran;Jee, Young-Mee;Ko, Jae-Sung;Seo, Jeong-Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.12 no.2
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    • pp.183-193
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    • 2009
  • Purpose: Rotaviruses, noroviruses, astroviruses, and enteric adenoviruses cause acute gastroenteritis (AGE) in children. Some children with AGE have afebrile convulsions associated with viral gastroenteritis. The purpose of this study was to detect and genotype viruses from children with AGE or benign infantile seizures associated with mild gastroenteritis (BIS-MG). Methods: Between August 2004 and June 2005, 311 children with AGE were included. Four viral agents, including rotavirus, norovirus, astrovirus, and adenovirus, were analyzed from stool specimens of each patient using the latex agglutination method, enzyme immunoassay, and reverse transcriptase polymerase chain reaction. Genotyping of each virus was performed in 217 of the 311 children. Results: Among 217 children (male, 121; female, 96; mean age, 20.6${\pm}$15.4 months), rotavirus was detected in 109 (50.2%), norovirus in 28 (12.9%), adenovirus in 13 (6.0%), and astrovirus in 2 children (0.9%). Genotyping of rotavirus revealed positive results in 97 children; P[8]G3 in 36, P[4]G2 in 21, P[6]G4 in 10, P[4]G4 in 9, P[8]G9 in 6, P[8]G1 in 6, P[4]G3 in 4, P[4]G9 in 3, and P[6]G2 in 2. Genotyping of norovirus showed GII-4 in 27 of 28 children and GII-6 in 1 child. Sixteen children were diagnosed with BIS-MG. Rotavirus was detected in 13 of 16 children with BIS-MG, and norovirus in 2 children. Genotyping of rotavirus detected in children with BIS-MG revealed P[8]G3 in 6 children, P[4]G2 in 2 children, and P[4]G9 in 1 child. Conclusion: Analysis of viruses from stool specimens indicates that both rotavirus and norovirus are the main viruses related to BIS-MG in children.

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Development of Assay Methods for Enterotoxin of Escherichia coli Employing the Hybridoma Technology (잡종세포종기법을 이용한 대장균의 장독소 측정법 개발)

  • Kim, Moon-Kyo;Cho, Myung-Je;Park, Kyung-Hee;Lee, Woo-Kon;Kim, Yoon-Won;Choi, Myung-Sik;Park, Joong-Soo;Cha, Chang-Yong;Chang, Woo-Hyun;Chung, Hong-Keun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.151-161
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    • 1986
  • In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

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Ammonia Volatilization from Coated Urea in Paddy Soil of Direct Seeding Rice Culture (벼 건답직파재배에서 피복요소 시용에 따른 암모니아 휘산)

  • Lee, Dong-Wook;Park, Ki-Do;Park, Chang-Young;Jeon, Weon-Tae;Son, Il-Soo;Park, Sung-Tae;Lee, Suk-Soon;Kang, Ui-Gum
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.6
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    • pp.328-333
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    • 2005
  • Ammonia volatilization is the major form of nitrogen (N) loss from flooded paddy soils and causes low N use efficiency. The effects of controlled release fertilizer (latex coated urea complex fertilizer, LCU) on reducing N loss by ammonia volatilization was measured comparing with urea in rice culture system of direct seeding on dry soil. In the treatment of urea, $NH_4-N$ concentration in surface water after flooding increased rapidly up to $8-10mg\;L^{-1}$ as affected by topdressing, while in the LCU treatment $NH_4-N$ concentration in surface water was less than $1mg\;L^{-1}$ during rice growing season. Relation of $NH_4-N$ concentration in surface water and ammonia volatilization was significant in urea treatment. The amount of ammonia volatilized from rice paddy of LCU treatment was $2.4-3.0kg\;ha^{-1}$ and the rate of ammonia volatilization from N fertilizer applied was only 2.0-2.3% compared with 5.9-7.9% in urea treatment. Therefore, N loss by ammonia volatilization could be reduced by 72-76% with by LCU compared with urea in rice culture system of direct seeding on dry soil.

Ammonia Volatilization from Coated Urea in Paddy Soil of Transplanting Rice Culture (벼 이앙재배에서 피복요소 시용에 따른 암모니아 휘산)

  • Lee, Dong-Wook;Park, Ki-Do;Park, Chang-Young;Kang, Ui-Gum;Son, Il-Soo;Yun, Eul-Soo;Park, Sung-Tae;Lee, Suk-Soon
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.6
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    • pp.321-327
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    • 2005
  • Ammonia ($NH_3$) volatilization was measured from latex coated urea (LCU) and normal urea treated rice paddy under transplanting rice culture in Milyang in 2002 and 2003. The $NH_3$ volatilization from incubation experiment was significantly related with ammonium-N ($NH_4-N$) concentration and pH in the surface water. The correlation coefficients of $NH_3$ volatilization compared to the $NH_4-N$ and pH in surface water were significantly higher in urea than LCU. The $NH_3$ volatilization from both urea and LCU treatments was not increased in surface water of pH less than 8.0, while $NH_3$ volatilization increased significantly in the surface water of pH over 8.0. The results in the field experiment indicated that $NH_3$ volatilization after top-dressing of urea increased rapidly with increasing $NH_4-N$ concentration in soil and floodwater, and highest from 7 to 10 days after top-dressing. The amount of $NH_3$ volatilized from urea treatment was in the range of $4.9-8.4kg\;N\;ha^{-1}$. The variations of $NH_3$ volatilization in 2002 and 2003 were caused by changed N dynamics due to the different weather conditions such as rainfall and temperature. The amount of $NH_3$ volatilized from LCU treatment was significantly reduced compared to that of urea. The reason for the reduced $NH_3$ volatilization in LCU treatment would be due to the lower concentration of $NH_4-N$ in floodwater. The amount of $NH_3$ volatilized from LCU treated rice paddy was in the range of $1.2-1.8kg\;N\;ha^{-1}$, and the loss of N by ammonia volatilization was 2.0-2.3%. Loss of N by $NH_3$ volatilization with LCU treatment was reduced by 75-79% comparing to urea treatment.

Recommendation Rate of Slow Release Nitrogen Fertilizer Application for Early Dry Seeding Culture of Rice in Honam Plain Area (호남평야지에서 벼 건답직파 조기파종재배에 알맞은 완효성 질소비료 시비량)

  • Kim, Sang-Su;Choi, Weon-Yeong;Back, Nam-Hyun;Choi, Min-Gyu;Park, Hong-Kyu;Nam, Jeong-Kweon
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.5
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    • pp.292-297
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    • 2006
  • A research was carried out to investigate the release pattern of slow release nitrogen fertilizer compound(SRNC) that is latex coated urea(LCU) and to recommend the application rate of the fertilizer at dry seeding rice culture in Honam plain area. The experiment was conducted at experimental field(Jeonbug series, fine silty, mixed, nonacid, mesic family of Fluvaquentic Endoaquepts) of National Honam Agricultural Research Institute. A medium late maturing rice variety (Dongjinbyeo) was seeded at the rate of $60kg\;ha^{-1}$ by drill seeder on April 1, 1997 and March 30, 1998. Fertilizer application rate was $160-90-110kg\;ha^{-1}$ for $N-P_2O_5-K_2O$. Soil ammonium nitrate($NH_4-N$) contents remained higher in all SRNC plots than conventional ones and higher with the increase of SRNC application rate until panicle formation stage, but the contents was higher in the conventional than NRNC plots at the heading stage. The plant hight was taller in SRNC than conventional plots until maximum tiller stage, but became similar in both conventional and SRNC plots at heading stage. The culm and panicle number was greater in SRNC than conventional plots throughout all growth stage, but the effective tiller rate was higher in conventional ones. Nitrogen efficiency was higher in the SRNC than conventional plots, but the efficiency was decreasing with the higher SRNC level. The spikelet number per unit area was greater in SRNC than conventional plots, and increased with higher SRNC level. The more spikelet number with higher NRNC level resulted in lower 1,000-grain weight. The rice yield in conventional plot was similar with only 60% SRNC level, but lower than 80% and 100% SRNC levels. However, slight lodging was observed in 100% SRNC level. In conclusions, we recommend NRNC application level as 80% of standard nitrogen application for early dry seeding culture of rice Honam plain area.