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Flaviviruses Induce Pro-inflammatory and Anti-inflammatory Cytokines from Murine Dendritic Cells through MyD88-dependent Pathway

  • Aleyas, Abi G.;George, Junu A.;Han, Young-Woo;Kim, Hye-Kyung;Kim, Seon-Ju;Yoon, Hyun-A;Eo, Seong-Kug
    • IMMUNE NETWORK
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    • v.7 no.2
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    • pp.66-74
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    • 2007
  • Background: The genus Flavivirus consists of many emerging arboviruses, including Dengue virus (DV), Japanese encephalitis virus (JEV) and West Nile virus (WNV). Effective preventive vaccines remain elusive for these diseases. Mice are being increasingly used as the animal model for vaccine studies. However, the pathogenic mechanisms of these viruses are not clearly understood. Here, we investigated the interaction of DV and JEV with murine bone marrow-derived dendritic cells (bmDC). Methods: ELISA and FACS analysis were employed to investigate cytokine production and phenotypic changes of DCs obtained from bone marrow following flavivirus infection. Results: We observed that these viruses altered the cytokine profile and phenotypic markers. Although both viruses belong to the same family, JEV-infected bmDC produced anti-inflammatory cytokine (IL-10) along with pro-inflammatory cytokines, whereas DV infection induced production of large amounts of pro-inflammatory cytokines (IL-6 and TNF-${\alpha}$) and no IL-10 from murine bmDCs. Both flaviviruses also up-regulated the expression of co-stimulatory molecules such as CD40, CD80 and CD86. JEV infection led to down-regulation of MHC II expression on infected bmDCs. We also found that cytokine production induced by JEV and DV is MyD88-dependent. This dependence was complete for DV, as cytokine production was completely abolished in the absence of MyD88. With regard to JEV, the absence of MyD88 led to a partial reduction in cytokine levels. Conclusion: Here, we demonstrate that MyD88 plays an important role in the pathogenesis of flaviviruses. Our study provides insight into the pathogenesis of JEV and DV in the murine model.

Seedling Conditions for Kimchi Cabbage, Head Lettuce, Cabbage and Broccoli for a Riding-type Transplanter

  • Lim, Jung-Hyun;Park, Suh-young;Chae, Won-Byung;Kim, Sung-Kyeom;Choi, Seung-Kook;Yang, Eun-Young;Lee, Min-Ji;Park, Gyeong-Bin;Jang, Yoon-ah;Seo, Myeong-Hoon;Jang, Suk-Woo
    • Journal of Biosystems Engineering
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    • v.42 no.2
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    • pp.104-111
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    • 2017
  • Purpose: We have studied the necessary qualities of seedlings of some leafy vegetables for a riding-type two-row automatic transplanter. When seedlings are planted using a transplanter, long roots may be rounded in the bottom of the tray, and this can interfere with the separation of the seedlings. Uprightness related to leaf spread angle is an important quality for seedlings in mechanical planting. Methods: To select cultivars suitable for the transplanter, we compared varieties of Kimchi cabbage (Chukwang, Daetong, Whipalam and Namdo), head lettuce (Abi and Sensation), cabbage (YR Onnuri, YR Hogel, Harutama, and Ogane), and broccoli (Nicegreen and Earlyyou). To compare the effect of bed soil on root formation and growth, we used five types of soil: Chologi, Burger, Wonjomix, Bio, and Baroker with 2.6-3 L per tray. Growth increment and the degree of root formation were measured according to the RDA guidelines 25 days after sowing for Kimchi cabbage and head lettuce and 44 days after sowing for cabbage and broccoli. Conclusions: According to the plug tray, the optimum seedling age in both 128- and 200-hole trays was 28 days for Kimchi cabbage and 44 days for cabbage and broccoli. Head lettuce took 35 days in 128-hole trays and 31 days in 200-hole trays. Burger soil was most effective for root formation and growth of the four kinds of leafy vegetables; it appeared that smaller soil volume led to faster root formation.

Studies on Proteolytic and Fibrinolytic Activity of Bacillus subtilis JM-3 Isolated from Anchovy Sauce (멸치액젓으로부터 분리한 Bacillus subtilis JM-3의 단백질 분해활성과 혈전 용해 활성에 관한 연구)

  • Lee, Sang-Soo;Kim, Sang-Moo;Park, Uk-Yeon;Kim, Hee-Yun;Shin, Il-Shik
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.283-289
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    • 2002
  • This study was performed to search for potential microorganism that has rapid fermenting and physiological function from anchovy sauce. We isolated three bacterial strains, JM-1, JM-2, and JM-3 with proteolytic and fibrinolytic activity from anchovy sauce. Among the 3 bacterial strains, JM-3 showed the strongest proteolytic and fibrinolytic activity. Bacterial strain JM-3 was gram-positive rod, motile and formed endospore. The 16S rRNA of bacterial strain JM-3 was amplified by PCR and then its sequence was determined by ABI 310 genetic analyzer. The 16S rRNA sequence of bacterial strain JM-3 was compared to BLAST DNA database and identified to Bacillus subtilis with 99% of homology. The optimum temperature, pH and NaCl concentration for growth of B. subtilis JM-3 were $40^{\circ}C$, 5.0 and 0%, respectively. The optimum temperature, pH and NaCl concentration for proteolytic and fibrinolytic enzyme production of B. subtilis JM-3 were same as optimum conditions for growth. At 20% of NaCl concentration which is common NaCl concentration of fish sauce, B. subtilis JM-3 showed about 60% of proteolytic and fibrinolytic activity of 0% NaCl concentration. From above results, we found that B. subtilis JM-3 will be able to used for starter of functional fish sauce.

SNPchaser : A Web-based Program for Detecting SNPs Substitution and Heterozygosity Existence (SNPchaser : DNA서열의 SNPs 치환 및 Heterozygosity 확인 프로그램)

  • Jang, Jin-Woo;Lee, Hyun-Chul;Lee, Myung-Hoon;Choi, Yeon-Shik;Choo, Dong-Won;Park, Kie-Jung;Lee, Dae-Sang
    • KSBB Journal
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    • v.24 no.4
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    • pp.410-414
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    • 2009
  • Single-nucleotide polymorphisms (SNPs) are the DNA sequences difference among the same species in the level of nucleic acids and are widely applied in clinical fields such as personalized medicine. The routine and labor-intensive methods to determine SNPs are performing the sequence homology search by using BLAST and navigating the trace of chromatogram files generated by high-throughput DNA sequencing machine by using Chromas program. In this paper, we developed SNPchaser, a web-based program for detecting SNPs substitution and heterozygosity existence, to improve the labor-intensive method in determining SNPs. SNPchaser performed sequence alignment and visualized the suspected region of SNPs by using user's reference sequence, AB1 files, and positional information of SNPs. It simultaneously provided the results of sequences alignment and chromatogram of relevant area of SNPs to user. In addition, SNPchaser can easily determine existence of heterozygosity in SNPs area. SNPchaser is freely accessible via the web site http://www.bioinformatics.ac.kr/SNPchaser and the source codes are available for academic research purpose.

No Associations between Schizophrenia and D22S280 Marker on Synapsin III Gene in Korean Males (한국인 남자에서 Synapsin III 유전자의 D22S280 표지자와 정신분열병의 연합연구)

  • Lee, Yu-Sang;Park, Chong-Won;Lee, Seung-Yeoun;Lee, Suk-Jin;Park, Yong-Bum;Shin, Yoon-Sik;Yoo, Jang-Keun;Hong, Kyung Sue;Yang, Byung-Hwan
    • Korean Journal of Biological Psychiatry
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    • v.13 no.4
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    • pp.260-266
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    • 2006
  • Objectives : Synapsin III near VCFS region on chromosome 22q affects. It could be an interesting candidate gene for schizophrenia. D22S280 is a highly polymorphic genetic marker residing in synapsin III. We examined association of D22S280 marker on synapsin III with Korean patients with schizophrenia. Methods : The subjects were 46 male Korean patients with schizophrenia and 60 male normal controls. Using polymerase chain reaction, gel electrophoresis, ABI 310 genetic analyzer, and GeneScan Collection 3.1 software, we confirmed genotypes of D22S280 marker. We examined Hardy-Weinberg equilibrium and case-control association using SAS/Genetic 9.1.3. Results : Genotypes of both schizophrenia and control groups were in Hardy-Weinberg equilibrium. We could not find any significant statistical differences in allele-wise(${\chi}^2$=10.4, df=6, p=0.098) and genotype-wise (${\chi}^2$=22.1 df=19, p=0.258) analyses of D22S280 marker between schizophrenia and normal controls. Individual allele analyses with df=1 showed significant differences in A1(p=0.025) and A7(p=0.034) allele, which were not significant following Bonferroni corrections(A1:p=0.177, A7:p=0.235). Conclusion : We couldn't find any association between schizophrenia and the synapsin III gene. Given the small number of subjects studied, further investigations are needed.

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Multiple Alternating Immunizations with DNA Vaccine and Replication-incompetent Adenovirus Expressing gB of Pseudorabies Virus Protect Animals Against Lethal Virus Challenge

  • Kim, Seon-Ju;Kim, Hye-Kyung;Han, Young-Woo;Aleyas, Abi G.;George, Junu A.;Yoon, Hyun-A;Yoo, Dong-Jin;Kim, Koan-Hoi;Eo, Seong-Kug
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1326-1334
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    • 2008
  • The prime-boost vaccination with DNA vaccine and recombinant viral vector has emerged as an effective prophylactic strategy to control infectious diseases. Here, we compared the protective immunities induced by multiple alternating immunizations with DNA vaccine (pCIgB) and replication-incompetent adenovirus (Ad-gB) expressing glycoprotein gB of pseudorabies virus (PrV). The platform of pCIgB-prime and Ad-gB-boost induced the most effective immune responses and provided protection against virulent PrV infection. However, priming with pCIgB prior to vaccinating animals by the DNA vaccine-prime and Ad-boost protocol provided neither effective immune responses nor protection against PrV. Similarly, boosting with Ad-gB following immunization with DNA vaccine-prime and Ad-boost showed no significant responses. Moreover, whereas the administration of Ad-gB for primary immunization induced Th2-type-biased immunity, priming with pCIgB induced Th1-type-biased immunity, as judged by the production of PrV-specific IgG isotypes and cytokine IFN-$\gamma$. These results indicate that the order and injection frequency of vaccine vehicles used for heterologous prime-boost vaccination affect the magnitude and nature of the immunity. Therefore, our demonstration implies that the prime-boost protocol should be carefully considered and selected to induce the desired immune responses.

Detection of infectious canine hepatitis virus by TaqMan real-time PCR method (TaqMan 실시간 PCR법에 의한 개 전염성 간염 바이러스의 검출)

  • Wang, Hye-young;Choi, Jae-yong;Lee, Mi-jin;Park, Jin-ho;Cho, Mae-Rim;Han, Jae-cheol;Choi, Kyoung-seong;Chae, Joon-seok
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.655-662
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    • 2004
  • The aim of this work was the validation of a rapid real-time PCR assay based on TaqMan technology for the unequivocal identification of infectious canine hepatitis (ICH) virus, to be used directly on DNA purified from blood specimens. A real-time PCR system targeting at the E3 ORFA gene sequence of canine adenovirus type 1 was optimized and validated through comparative analysis of samples using conventional PCR system. The real-time PCR assay based on TaqMan technology could disclose 23 (37.7%) out of 61 samples as PCR positive. In contrast, 18 (29.5%) samples were found PCR positive when conventional PCR was applied on these samples. The use of the ABI Prism 7700 sequence detection system allowed the efficient determination of the amplified product accumulation through a fluorogenic probe. The entire real-time TaqMan PCR assay, including DNA extraction, amplification, and detection could be completed within 3 hours. The detection method of real-time TaqMan PCR assay was 1,000 times more sensitive than conventional PCR. Real-time TaqMan probe and primer set developed and optimized in this study is a sensitive, rapid and accurate method for detection of ICH virus and can be effective screening tool for the detection of ICH in a diagnostic laboratory routines.

Molecular Survey of Latent Pseudorabies Virus Infection in Nervous Tissues of Slaughtered Pigs by Nested and Real-time PCR

  • Yoon Hyun A;Eo Seong Kug;Aleyas Abi George;Park Seong Ok;Lee John Hwa;Chae Joon Seok;Cho Jeong Gon;Song Hee Jong
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.430-436
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    • 2005
  • In this study, the prevalence and quantity of a latent pseudorabies virus (PrV) infection in the nervous tissues of randomly selected pigs was determined via nested and real-time PCR. The nervous tissues, including the trigeminal ganglion (TG), olfactory bulb (OB), and brain stem (BS), were collected from the heads of 40 randomly selected pigs. The majority of the nervous tissues from the selected pigs evidenced a positively amplified band on nested PCR. In particular, nested PCR targeted to the PrV glycoprotein B (gB) gene yielded positive results in all of the BS samples. Nested PCR for either the gE or gG gene produced positive bands in a less number of nervous tissues ($57.5\%$ and $42.5\%$, respectively). Real-time PCR revealed that the examined tissues harbored large copy numbers of latent PrV DNA, ranging between $10^{0.1}\;and\;10^{7.2}(1-1.58{\times}10^7)$ copies per $1{\mu}g$ of genomic DNA. Real-time PCR targeted to the PrV gE gene exhibited an accumulated fluorescence of reporter dye at levels above threshold, thereby indicating a higher prevalence than was observed on the nested PCR ($100\%$ for BS, $92\%$ for OB, and $85\%$ for TG). These results indicate that a large number of farm-grown pigs are latently infected with a field PrV strain with a variety of copy numbers. This result is similar to what was found in association with the human herpes virus.

Short Bowel Syndrome as the Leading Cause of Intestinal Failure in Early Life: Some Insights into the Management

  • Goulet, Olivier;Nader, Elie Abi;Pigneur, Benedicte;Lambe, Cecile
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.22 no.4
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    • pp.303-329
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    • 2019
  • Intestinal failure (IF) is the critical reduction of the gut mass or its function below the minimum needed to absorb nutrients and fluids required for adequate growth in children. Severe IF requires parenteral nutrition (PN). Pediatric IF is most commonly due to congenital or neonatal intestinal diseases or malformations divided into 3 groups: 1) reduced intestinal length and consequently reduced absorptive surface, such as in short bowel syndrome (SBS) or extensive aganglionosis; 2) abnormal development of the intestinal mucosa such as congenital diseases of enterocyte development; 3) extensive motility dysfunction such as chronic intestinal pseudo-obstruction syndromes. The leading cause of IF in childhood is the SBS. In clinical practice the degree of IF may be indirectly measured by the level of PN required for normal or catch up growth. Other indicators such as serum citrulline have not proven to be highly reliable prognostic factors in children. The last decades have allowed the development of highly sophisticated nutrient solutions consisting of optimal combinations of macronutrients and micronutrients as well as guidelines, promoting PN as a safe and efficient feeding technique. However, IF that requires long-term PN may be associated with various complications including infections, growth failure, metabolic disorders, and bone disease. IF Associated Liver Disease may be a limiting factor. However, changes in the global management of IF pediatric patients, especially since the setup of intestinal rehabilitation centres did change the prognosis thus limiting "nutritional failure" which is considered as a major indication for intestinal transplantation (ITx) or combined liver-ITx.

Effects of Low Intensity Combined Exercise Training with Blood Flow Restriction on Body Composition and Cardiovascular Responses in Elderly Females (저강도 혈류제한 복합운동이 여성노인들의 신체조성과 심혈관 요인들에 미치는 영향)

  • Kim, Daeyeol;Kuk, Doohong;Park, Hyeok
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.1
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    • pp.362-370
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    • 2019
  • This study was conducted to investigate effects of 12 weeks of combined exercise training with blood flow restriction (BFR) on body composition (weight, %body fat, lean body mass, body mass index (BMI)) and cardiovascular responses (brachial-ankle pulse wave velocity (ba PWV) and ankle-brachial index (ABI)) in elderly women. Participants (N = 43, Females) were randomly assigned into a combined exercise with BFR (n = 14, BFR), only combined exercise (n =14, EX) or non-exercise control group (n = 15, CON). Two-way repeated measures ANOVA with contrast testing was utilized for data analysis. Alpha was set at p < 0.05. Body composition (weight, %body fat, BMI) in BFR was significantly changed, and %body fat in EX was significantly decreased, but there was no change in the CON. In addition, the right and left ba PWV values in the BFR were significantly decreased, while only the left side ba PWV in EX was significantly decreased and there was no change in the CON. Moreover, the % change and effect size of most variables in the BFR were higher than the EX. Taken together, the results indicate that even though BFR and EX groups performed the same combined exercise training, BFR had additional stimulations of the sympathetic nerve system due to blood flow restriction. Thus, BFR training is more beneficial and has greater effects on body composition and cardiovascular responses in elderly females.