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Blastogenic responses of splenic Iymphocytes to Naegleria fowleri Iysates and T-cell mitogen in mice with primary amoebie meningoencephalitis. (실험적 뇌막수염에 있어 Naegleria fowleri 항원에 대한 세포매개성 면역 반응)

  • Park, Gwang-Min;Ryu, Jae-Suk;Im, Gyeong-Il
    • Parasites, Hosts and Diseases
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    • v.25 no.1
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    • pp.1-6
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    • 1987
  • This study was to observe the changes of blastogenic responses of splenic Iymphocytes to T-cell mitogens, N. fcwleri Iysate and concanaualin A, and serum antibody titer during the course of experimental PAM in mice. Naegleria fcwleri, strain 0359, was cultured in the CGVS medium axenically and inoculated intranasally with $7{\times}10^4$ trophozoites for the development of experimental PAM in mice. The amoebae were subjected to ultrasonication and centrifuged at 20,000g for 60 minutes, and filtered through $0.2{\mu\textrm{m}}$ filter membrane. The supernatant, N. fcwleri Iysate, was used as T-cell mitogen, and antigen for ELISA. The serum antibody was examined by ELISA using peroxidase conjugate. Two hundred ${\mi}l$ of $10^6$ splenocytes in RPMI 1640 containing 0% fetal calf serum were added to each well of a microtiter plate. To each well was added T-cell mitogens, $100{\mu}g/ml$ of N. fowleri Iysate or $4{\mu}g/ml$ of con. A, and the plates were incubated for 42 hours at $37^{\circ}C$ in 5% $CO_2$ incubator. Cultures were pulsed with of $methyl-(^3H)-thymidine$ 6 hour before harvesting. The mean blastogenic response of the splenocytes to N. fewleri Iysate was reduced, whereas that to con. A was also reduced up to on day 11 after infection. Both of these results were statistically significant compared with those of uninfected control group. The serum antibody titers were increased gradually up to day 15. The results indicated that there was an impairment of the blastogenic response of splenocytes to N. fowleri Iysate during the acute course of experimental PAM in mice.

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A Health Risk Assessment of Tributyltin Compounds in Fishes and Shellfishes in Korea. (국내 유통중인 어패류 섭취에 따른 유기주석화합물의 인체 위해성 평가)

  • Choi, Shi-Nai;Choi, Hye-Kyung;Song, Hoon;Oh, Chang-Hwan;Park, Jong-Sei
    • Journal of Food Hygiene and Safety
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    • v.17 no.3
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    • pp.137-145
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    • 2002
  • Tributyltin compounds have been increasingly used in the form of plastic stabilizers, catalytic agents, industrial agricultural biocides, antifouling paint, and pesticides. Among these organotin compounds, large amounts of tributyltin(TBT) and triphenyltin(TPT) have been used as antifouling agents because they have a superior ability to prevent marine organism from being encrusted on ship bottoms and in culturing nets. Environmental pollution by these organotin compounds in the aquatic environment were undertaken. The international maritime Organization's established a provisional tolerable daily intake(TDI) of 1.6[micro]g TBTO/kg/ B.W. The Food and Agiculture Organization (of the United Nations)/world Health Organization's (FAO/WHO) proposed a TDI of 0.5ug TPT/kg BW/d. This study is conducted monitoring of TBT on seafoods in Korea and risk assessment for exposure on TBT in seafoods. Total hazard index(using Reference Dose : 0.3 ug TBTO/kg B.W/day) of intake exposure on seafoods is 0.04 as the 50th percentile, 0.08 as the 95th percentile. This value is estimated by Monte-Carlo simulation using Crystal Ball(Decisioneering Co., 2001).

Microbiological and Immunological Investigation on the Bacteroides gingivalis in Rapidly Progressive and Adult Periodontitis in Korean (한국인 급성진행성 및 성인성 치주염의 원인균인 Bacteroides gingivalis에 대한 미생물 및 면역학적 연구)

  • Chung, Chong-Pyoung;Lee, Jong-Heun;Chung, Hyun-Ju
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.309-321
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    • 1987
  • For the investigation of microbiological and immunological specificity of Bacteroides gingivalis, Bacteroides gingivalis were isolated, enumerated and characterized from 13 Korean rapidly progressive periodontitis and 7 healthy control by anaerobic culture technique. The total proportion of black-pigmented Bacteroides from Korean R.P.P. patients and healthy control were 8.78% and 0.92%, respectively, among total isolated black-pigmented Bacteroides. In antibiotic susceptibility test, Bacteroides gingivalis isolated from R.P.P. patients were sensitive to Ampicillin and Tetracycline, and resistant to Gentamicin and Erythromycin in disc diffusion method. In antibiotic broth dilution method, the minimum inhibitory concentration(MIC) to Bacteroides gingivalis was 2 unit/ml of Penicillin and $0.25{\sim}1{\mu}g/ml$ of Tetracycline, respectively. The concentration of serum IgG in rapidly progressive periodontitis patients were sigificantly higher than that of healthy control, and concentration of diluted gingival crevicular IgG has not any significant differences between two groups. Serum and gingival crevicular IgG antibody to Bacteroides gingivalis were significantly higher titer in rapidly progressive periodontitis patients to compare with healthy control. The lipopolysaccharide profiles of 2 Korean B. gingivalis in silver stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis were similar to type strains of B. gingivalis and typical LPS band were appeared around the 24-Kd molecular weight. Immunodiffusion test and immunoelectrophoresis of the L.P.S. extracted from 2 Korean B. gingivalis and 2 kinds of type strains of B. gingivalis showed that B. gingivalis Korean-1 was reacted identically to B. gingivalis ATCC 33277. In trypsin and ${\alpha}$-glucosidase activity test of 2 Korean B. gingivalis, both of them revealed positive trypsin and negative ${\alpha}$-glucosidase activity, respectively. These investigation suggested that B. gingivalis is important pathogenic plaque bacteria for the pathogenesis of periodontitis and further study is needed to purify and characterize of the species-specific antigens of this organisms to develop monoclonal antibody and potential diagnostic reagents.

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A Clinical and Serologic Study of 21 Cases of Tsutsugamushi Disease Confirmed by Serologic test (혈청학적으로 확진된 쯔쯔가무시(Tsutsugamushi)병 21예에 대한 임상적 고찰)

  • Park, Jong-Seon;Kweon, Young-Su;Lee, Kwan-Ho;Hyun, Myung-Su;Chung, Moon-Kwan;Lee, Hyun-Woo
    • Journal of Yeungnam Medical Science
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    • v.7 no.1
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    • pp.151-163
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    • 1990
  • Tsutsugamushi disease is an acute febrile disease caused by Rickettsia tsutsugamushi, and which has been reported with increasing frequency through the nation since 1986. We experienced 21 cases of Tsutsugamushi disease diagnosed with serologic test occuring in Taegu city and Kyungpook province during October-November, 1989. The results of survey are as follow. 1) Of 21 cases, 12(57%) were males and 9(43%) were females, and the peak incidence was the 4th decade. 2) The outbreak was in October to November and the peak incidence was in October. 3) The most frequent symptoms were fever and chill(100%), myalgia(95%), headache(90%). Eschar and rash were observed in 18 patients(86%) and the eschar was detected in all over the body, especially thorax(33%) and lower extremity(22%). 4) Laboratory features were SGOT elevation(83%), SGPT elevation(61%), LDH elevation(67%). leukocytosis (38%). 5) Indirect immunofluorescent antibody test was done m 18 patients and the antibody titer was above 1 : 320 in all patients. 6) The chloramphenicol, tetracycline or doxycycline regimens were very effective and mean duration of defervescence from iniation of therapy was 1.3 days. 7) The complication such as meningitis or shock, was not seen.

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Effect of 2-2'-dipyridyl in culture media and combined advantage of Streptococcus parauberis vaccine for preparation of Vibrio harveyi vaccine on olive flounder, Paralichthys olivaceus (Vibrio harveyi 백신의 효능 향상을 위한 배양 배지내 2-2'-dipyridyl 첨가 및 연쇄구균 백신과 혼합 투여의 효과)

  • Kim, Myoung Sug;Jung, Sung Hee;Hong, Suhee
    • Journal of Fisheries and Marine Sciences Education
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    • v.26 no.6
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    • pp.1366-1372
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    • 2014
  • This study was conducted to select the media for the formalin killed vaccine (FKC) production of Vibrio harveyi and its application for olive flounder, Paralichthys olivaceus. For this, we have investigated the immune effects of Vibrio harveyi FKC vaccines grown in 3 different media i.e. Tryptic Soy Broth (TSB), TSB containing 2-2'-dipyridyl (TSB-D), Brain Heart Infusion Broth (BHIB) on the production of agglutinating antibody and protection against Vibrio harveyi in olive flounder. Additionally, a dual vaccine was prepared by combining Streptococcus parauberis vaccine to V. harveyi vaccine and its efficacy was also analyzed with the determination of optimal administration dosage. Consequently, olive flounder immunized with FKC grown in TSB-D showed the same protection with the vaccine grown in BHIB and the optimal dose of the vaccine was 10mg/kg of body weight. Indeed the dual vaccine showed higher agglutination titer and protection than control fish. The optimal dose for dual vaccine was 10mg/kg body weight for each of two vaccines.

Development of an Equine Antitoxin by Immunizing the Halla Horse with the Receptor-Binding Domain of Botulinum Neurotoxin Type A1

  • Kim, Na Young;Park, Kyung-eui;Lee, Yong Jin;Kim, Yeong Mun;Hong, Sung Hyun;Son, Won Rak;Hong, Sungyoul;Lee, Saehyung;Ahn, Hye Bin;Yang, Jaehyuk;Seo, Jong-pil;Lim, Yoon-Kyu;Yu, Chi Ho;Hur, Gyeung Haeng;Jeong, Seong Tae;Lee, Hun Seok;Song, Kyoung;Kang, Tae Jin;Shin, Young Kee;Choi, Joon-Seok;Choi, Jun Young
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1165-1176
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    • 2019
  • Botulinum neurotoxins (BoNTs), produced by Clostridium botulinum, are the most toxic substances known. However, the number of currently approved medical countermeasures for these toxins is very limited. Therefore, studies on therapeutic antitoxins are essential to prepare for toxin-related emergencies. Currently, more than 10,000 Halla horses, a crossbreed between the native Jeju and Thoroughbred horses, are being raised in Jeju Island of Korea. They can be used for equine antitoxin experiments and production of hyperimmune serum against BoNT/A1. Instead of the inactivated BoNT/A1 toxoid, Halla horse was immunized with the receptor-binding domain present in the C-terminus of heavy chain of BoNT/A1 (BoNT/A1-HCR) expressed in Escherichia coli. The anti-BoNT/A1-HCR antibody titer increased rapidly by week 4, and this level was maintained for several weeks after boosting immunization. Notably, $20{\mu}l$ of the week-24 BoNT/A1-HCR(-immunized) equine serum showed an in vitro neutralizing activity of over 8 international units (IU) of a reference equine antitoxin. Furthermore, $20{\mu}l$ of equine serum and $100{\mu}g$ of purified equine $F(ab^{\prime})_2$ showed 100% neutralization of 10,000 $LD_{50}$ in vivo. The results of this study shall contribute towards optimizing antitoxin production for BoNT/A1, which is essential for emergency preparedness and response.

Escherichia coli-Derived Outer Membrane Vesicles Deliver Galactose-1-Phosphate Uridyltransferase and Yield Partial Protection against Actinobacillus pleuropneumoniae in Mice

  • Quan, Keji;Zhu, Zhuang;Cao, Sanjie;Zhang, Fei;Miao, Chang;Wen, Xintian;Huang, Xiaobo;Wen, Yiping;Wu, Rui;Yan, Qigui;Huang, Yong;Ma, Xiaoping;Han, Xinfeng;Zhao, Qin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2095-2105
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    • 2018
  • In our previous studies, we have identified several in vivo-induced antigens and evaluated their potential as subunit vaccine candidates in a murine model, in which the recombinant protein GalT showed the most potent immunogenicity and immunoprotective efficacy against Actinobacillus pleuropneumoniae. To exploit a more efficient way of delivering GalT proteins, in this study, we employed the widely studied E. coli outer membrane vesicles (OMVs) as a platform to deliver GalT protein and performed the vaccine trial using the recombinant GalT-OMVs in the murine model. Results revealed that GalT-OMVs could elicit a highly-specific, IgG antibody titer that was comparable with the adjuvant GalT group. Significantly higher lymphocyte proliferation and cytokines secretion levels were observed in the GalT-OMVs group. 87.5% and 50% of mice were protected from a lethal dose challenge using A. pleuropneumoniae in active or passive immunization, respectively. Histopathologic and immunohistochemical analyses showed remarkably reduced pathological changes and infiltration of neutrophils in the lungs of mice immunized with GalT-OMVs after the challenge. Taken together, these findings confirm that OMVs can be used as a platform to deliver GalT protein and enhance its immunogenicity to induce both humoral and cellular immune responses in mice.

Biological and molecular characterization of feline caliciviruses isolated from cats in South Korea

  • Yang, Dong-Kun;Park, Yu-Ri;Yoo, Jae Young;Choi, Sung-Suk;Park, Yeseul;An, Sungjun;Park, Jungwon;Kim, Heui-Jin;Kim, Jongho;Kim, Ha-Hyun;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
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    • v.60 no.4
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    • pp.195-202
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    • 2020
  • Feline calicivirus (FCV) infection results in a common upper respiratory disease associated with oral ulceration in cats. Although FCV infection has been reported in cats worldwide, the biologic and genetic features of South Korean FCV are unclear. We aimed to investigate the biological and genetic features of South Korean FCV isolates. Crandell-Rees feline kidney (CRFK) cells were used to isolate FCV from 58 organ homogenate samples. The FCV isolates were confirmed by cytopathic effects, immunofluorescence, electron microscopy, and reverse transcription polymerase chain reaction assays. Viral genetic analysis was carried out with VP2 gene and complete genomes of FCVs. Five viruses propagated in CRFK cells were confirmed to be FCVs. The FCV17D283 isolate showed the highest viral titer of 107.2 TCID50/mL at 36 h post-inoculation. Korean FCV isolates did not grow well in Vero, BHK-21, A72, or Madin-Darby canine kidney cells. The FCV17D03 and FCV17D283 isolates had the highest genetic similarity (80.1% and 86.9%) with the UTCVM-H1 and 14Q315 strains, which were isolated in the United States and South Korea in 1995 and 2014, respectively. We isolated five FCVs from cats and detected important genetic differences among them. FCV isolates did not show any virulent effects in mice.

Efficacy of Living Donor Liver Transplantation in Patients with Methylmalonic Acidemia

  • Jang, Jae Guk;Oh, Seak Hee;Kim, Yu Bin;Kim, Seo Hee;Yoo, Han-Wook;Lee, Beom Hee;Namgoong, Jung-Man;Kim, Dae Yeon;Kim, Ki-Hun;Song, Gi-Won;Moon, Deok-Bog;Hwang, Shin;Lee, Sung-Gyu;Kim, Kyung Mo
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.24 no.3
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    • pp.288-294
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    • 2021
  • Purpose: Despite aggressive medical and nutritional management, patients with methylmalonic acidemia (MMA) often suffer from multi-organ damage. Early deceased donor liver transplantation (DDLT) has emerged as an intervention to prevent disease progression. We investigated the efficacy of living donor LT (LDLT) with a potential carrier of MMA and a small volume of graft in patients with MMA as an alternative to DDLT. Methods: Of five patients (three male, two female; median age 5.7 years; range, 1.3-13.7 years), four underwent carrier LDLT, while one underwent non-carrier auxiliary LDLT. All patients received pre- and post-LT continuous renal replacement therapy and were provided with minimal restriction diet according to serum MMA level after LT. MMA levels in the serum and urine, the incidence of metabolic crisis, and clinical findings before and after LT were compared. Results: The survival rate was 100% during 2.2 years of follow up period after LT. In all five cases, MMA titer in the serum after transplantation decreased with less restrictive diet. Metabolic crisis was not observed during the follow-up period. In addition, no patient showed progression of severe renal impairment requiring hemodialysis. Progression of delayed cognitive development was not observed. Social functioning with improved neuropsychiatric development was observed. Conclusion: This study showed that LDLT achieved improved quality of life with less restrictive diet, therefore it could be a feasible alternative option to DDLT for the treatment of patients with MMA, even with an auxiliary LT.

Age-dependent immune response in pigs against foot-and-mouth disease virus in vitro

  • Roh, Jae-Hee;Bui, Ngoc Anh;Lee, Hu Suk;Bui, Vuong Nghia;Dao, Duy Tung;Vu, Thanh Thi;Hoang, Thuy Thi;So, Kyoung-Min;Yi, Seung-Won;Kim, Eunju;Hur, Tai-Young;Oh, Sang-Ik
    • Journal of Animal Science and Technology
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    • v.63 no.6
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    • pp.1376-1385
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    • 2021
  • Foot-and-mouth disease, one of the most contagious diseases in cloven-hoofed animals, causes significant economic losses. The pathogenesis of foot-and-mouth disease virus (FMDV) infection is known to differ with age of the animals. In this study, we aimed to reveal the difference in immunological response in the initial stage of FMDV infection between piglets and adult pigs. Peripheral blood mononuclear cells (PBMCs) were isolated from 3 piglets (8 weeks old) and 3 pigs (35 weeks old) that were not vaccinated against FMDV. O-type FMDV (2 × 102 median tissue culture infectious dose) was inoculated into porcine PBMCs and the cells were incubated at 37.0℃ under 5% CO2 for various time periods (0, 1, 3, 6, 12, 24, and 48 h). The total RNA was obtained from the FMDV-inoculated PBMCs after each time point, and the virus titer was investigated in these RNA samples. Furthermore, dynamics of mRNA expression of the six tested cytokines (interferon [IFN]-α, IFN-γ, interleukin [IL]-6, IL-8, IL-10, and tumor necrosis factor [TNF]-α) in FMDV-inoculated porcine PBMCs were evaluated by time-series analysis to determine the differences, if any, based on the age of the pigs. The PBMCs of piglets contained the highest quantity of FMDV mRNA at 6 hours post-inoculation (hpi), and the PBMCs of pigs had the highest quantity of FMDV mRNA at 3 hpi. The mean cycle threshold-value in the PBMCs steadily decreased after the peak time point in the piglets and pigs (6 and 3 hpi, respectively). The dynamics of mRNA expression of all cytokines except TNF-α showed age-dependent differences in FMDV-inoculated PBMCs. The mRNA expression of most cytokines was more pronounced in the piglets than in the pigs, implying that the immune response against FMDV showed an age-dependent difference in pigs. In conclusion, within 48 hpi, the 8-week-old piglets responded more rapidly and were more sensitive to FMDV infection than the 35-week-old pigs, which could be associated with the difference in the pathogenesis of FMDV infection among the pigs. These results provide valuable insights into the mechanisms underlying the age-dependent differences in immune response in pigs against FMDV infection.