• Title/Summary/Keyword: dsRNA

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Toll-like Receptor3-mediated Induction of Chemokines in Salivary Epithelial Cells

  • Li, Jingchao;Jeong, Mi-Young;Bae, Ji-Hyun;Shin, Yong-Hwan;Jin, Meihong;Hang, Sung-Min;Lee, Jeong-Chai;Lee, Sung-Joong;Park, Kyung-Pyo
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.4
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    • pp.235-240
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    • 2010
  • Toll-like receptors (TLRs) functionally expressed in salivary epithelial cells, but their roles remain elusive. Among TLRs family, TLR3 is activated by dsRNA, a byproduct of viral infection. The aim of this study was to investigate the role of TLR3 in the inflammatory immune responses using HSG cells. Reverse transcriptase-polymerase chain reaction (RT-PCR), real-time PCR and ELISA were performed to identify expression of TLRs and TLR3-mediated chemokine inductions. The chemotaxis assay of activated T lymphocytes was also performed. Treatment of HSG cells with polyinosinic: polycytidylic acid (poly(I:C)) significantly increased interferon-$\gamma$-inducible protein 10 (IP-10), interferoninducible T-cell $\alpha$ chemoattractant (I-TAC), and regulated on activation, normal T-cells expressed and secreted (RANTES) gene expressions in a concentration-dependent manner. Anti-TLR3 antibody blocked the increases of IP-10 and I-TAC genes. Poly(I:C)-induced increases of IP-10 and I-TAC were also confirmed at protein levels from cell lysates, but their release into extracellular medium was detected only in IP-10. We found that the culture media from HSG cells stimulated with poly(I:C) significantly increases T lymphocyte migration. Our results suggest that TLR3 plays an important role in chemokine induction, particularly IP-10, in salivary epithelial cells.

Characterization of Melon necrotic spot virus Occurring on Watermelon in Korea

  • Kwak, Hae-Ryun;Kim, Jeong-Soo;Cho, Jeom-Deog;Lee, Joong-Hwan;Kim, Tae-sung;Kim, Mi-Kyeong;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • v.31 no.4
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    • pp.379-387
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    • 2015
  • Melon necrotic spot virus (MNSV) was recently identified on watermelon (Citrullus vulgaris) in Korea, displaying as large necrotic spots and vein necrosis on the leaves and stems. The average occurrence of MNSV on watermelon was found to be 30-65% in Hapcheon and Andong City, respectively. Four isolates of the virus (MNSV-HW, MNSV-AW, MNSV-YW, and MNSV-SW) obtained from watermelon plants in different areas were non-pathogenic on ten general indicator plants, including Chenopodium quinoa, while they infected systemically six varieties of Cucurbitaceae. The virus particles purified by 10-40% sucrose density gradient centrifugation had a typical ultraviolet spectrum, with a minimum at 245 nm and a maximum at 260 nm. The morphology of the virus was spherical with a diameter of 28-30 nm. Virus particles were observed scattered throughout the cytoplasm of watermelon cells, but no crystals were detected. An ELISA was conducted using antiserum against MNSV-HW; the optimum concentrations of IgG and conjugated IgG for the assay were $1{\mu}l/ml$ and a 1:8,000-1:10,000 dilutions, respectively. Antiserum against MNSV-HW could capture specifically both MNSV-MN from melon and MNSV-HW from watermelon by IC/RT-PCR, and they were effectively detected with the same specific primer to produce product of 1,172 bp. The dsRNA of MNSV-HW had the same profile (4.5, 1.8, and 1.6 kb) as that of MNSV-MN from melon. The nucleotide sequence of the coat protein of MNSV-HW gave a different phylogenetic tree, having 17.2% difference in nucleotide sequence compared with MNSV isolates from melon.

Primary Cilium by Polyinosinic:Polycytidylic Acid Regulates the Regenerative Migration of Beas-2B Bronchial Epithelial Cells

  • Gweon, Bomi;Jang, Tae-Kyu;Thuy, Pham Xuan;Moon, Eun-Yi
    • Biomolecules & Therapeutics
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    • v.30 no.2
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    • pp.170-178
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    • 2022
  • The airway epithelium is equipped with the ability to resist respiratory disease development and airway damage, including the migration of airway epithelial cells and the activation of TLR3, which recognizes double-stranded (ds) RNA. Primary cilia on airway epithelial cells are involved in the cell cycle and cell differentiation and repair. In this study, we used Beas-2B human bronchial epithelial cells to investigate the effects of the TLR3 agonist polyinosinic:polycytidylic acid [Poly(I:C)] on airway cell migration and primary cilia (PC) formation. PC formation increased in cells incubated under serum deprivation. Migration was faster in Beas-2B cells pretreated with Poly(I:C) than in control cells, as judged by a wound healing assay, single-cell path tracking, and a Transwell migration assay. No changes in cell migration were observed when the cells were incubated in conditioned medium from Poly(I:C)-treated cells. PC formation was enhanced by Poly(I:C) treatment, but was reduced when the cells were exposed to the ciliogenesis inhibitor ciliobrevin A (CilioA). The inhibition of Beas-2B cell migration by CilioA was also assessed and a slight decrease in ciliogenesis was detected in SARS-CoV-2 spike protein (SP)-treated Beas-2B cells overexpressing ACE2 compared to control cells. Cell migration was decreased by SP but restored by Poly(I:C) treatment. Taken together, our results demonstrate that impaired migration by SP-treated cells can be attenuated by Poly(I:C) treatment, thus increasing airway cell migration through the regulation of ciliogenesis.

Distribution of Disease Severity and Group A Rotavirus Genotypes (VP7 & VP4) in Children with Acute Rotavirus Gastroenteritis (급성 로타바이러스 장염 소아에서 질환의 중증도와 로타바이러스 VP7 & VP4 유전형의 분포에 대한 연구)

  • Oh, Hyun-Ju;Kang, Hyun-Sik;Kang, Ki-Soo;Kim, Youn-Woo;Hong, Jung-Yun;Shin, Kyung-Sue;Lee, Jin-Sook;Lee, Su-Hyun;Lee, Keun-Hwa;Cho, Moon-Je;Suh, Dong-In
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.14 no.2
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    • pp.148-154
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    • 2011
  • Purpose: We aimed to study the distribution of rotavirus genotypes (VP7 and VP4) and disease severity of rotavirus gastroenteritis prevalent in our community. Methods: Stool samples were collected from 156 children who were hospitalized with rotavirus gastroenteritis from December 2007 to June 2008. The disease severity of all patients was scored using the Vesikari scale. After extraction of ds-RNA of the rotavirus, cDNA synthesis using reverse transcription and polymerase chain reaction (RT-PCR) and multiplex PCR was performed. Following this, the final identification of genotypes was performed. Results: Of the 156 samples, VP7(G) and VP4(P) genotypes were identified in 147 (94.2%) and 140 (89.7%) samples, respectively. G1 (116 of 147 samples; 78.9%) and P[8] (137 of 140 samples; 97.9%) were the most prevalent, respectively. Of the 138 samples identified of combination types of VP7 and VP4, G1P[8] (111 samples; 80.4%) was the most prevalent. Other combination types varied with very low distribution rates. 9.4% of genotypes were not included in the new vaccines. The disease severity score was $11.8{\pm}3.3$ ($mean{\pm}2SD$). The distribution of disease severity was mild or moderate in 37.8% and severe in 62.2% of patients. Conclusion: The most prevalent genotype combination of rotavirus was G1P[8] and genotypes not included in the vaccines represented 9.4% in our community. Disease severity distribution of hospitalized children with rotavirus gastroenteritis was higher in the severe than in the mild and moderate categories.

Dominant-species Variation of Soil Microbes by Temperate Change (온도변화에 기인한 토양미생물 우점종의 변화에 관한 연구)

  • Park, Kap-Joo;Lee, Byeong-Chol;Lee, Jae-Seok;Park, Chan-Sun;Cho, Myung-Hwan
    • Korean Journal of Environmental Biology
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    • v.29 no.1
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    • pp.52-60
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    • 2011
  • Today, the weather is changing continually, due to the progress of global warming. As the weather changes, the habitats of different organisms will change as well. It cannot be predicted whether or not the weather will change with each passing day. In particular, the biological distribution of the areas climate change affects constitutes a major factor in determining the natural state of indigenous plants; additionally, plants are constantly exposed to rhizospheric microorganisms, which are bound to be sensitive to these changes. Interest has grown in the relationship between plants and rhizopheric microorganisms. As a result of this interest we elected to research and experiment further. We researched the dominant changes that occur between plants and rhizospheric organisms due to global warming. First, we used temperature as a variable. We employed four different temperatures and four different sites: room temperature ($27^{\circ}C$), $+2^{\circ}C$, $+4^{\circ}C$, and $+6^{\circ}C$. The four different sites we used were populated by the following species: Pinus deniflora, Pinus koraiensis, Quercus acutissima, and Alnus japonica. We counted colonies of these plants and divided them. Then, using 16S rRNA analysis we identified the microorganisms. In conclusion, we identified the following genera, which were as follows: 10 species of Bacillus, 2 Enterobacter species, 4 Pseudomonas species, 1 Arthrobacter species, 1 Chryseobacterium species, and 1 Rhodococcus species. Among these genera, the dominant species in Pinus deniflora was discovered in the same genus, but a different species dominated at $33^{\circ}C$. Additionally, that of Pinus koraiensis changed in both genus and species which changed into the Chryseobacrterium genus from the Bacilus genus at $33^{\circ}C$.

Inhibitory Effects of Ginger and Beopje Ginger on DSS-induced Colitis in Mice (생강과 법제생강의 DSS(Dextran Sulfate Sodium)로 유도된 마우스의 대장염 억제 효과)

  • Kim, Sin-Jeong;Kim, So-Hee;Lim, Yaung-Iee;Kim, Yong-Gyu;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.4
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    • pp.477-484
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    • 2014
  • The purpose of this study was to examine the anti-inflammatory effects of ginger and processed (Beopje) ginger on colitis induced by 2.5% dextran sulfate sodium (DSS) in Balb/c mice. Beopje means a process that herbal medicines are treated by a specific Korean traditional method in order to obtain better pharmacological effects. Mice were fed saline or two different doses of ethanol extracts (ginger and processed (Beopje) ginger) once a day for 14 days. Colitis was induced from day 7 to 14 via administration of 2.5% DSS in drinking water. Experimental animals were divided into four groups: Nor (Normal, 200 ${\mu}L$ of saline without 2.5% DSS-treated group), Con (Control, 200 ${\mu}L$ of saline and 2.5% DSS treated group), G (500 mg/kg of ginger and 2.5% DSS treated group), and BG (500 mg/kg of Beopje ginger and 2.5% DSS treated group). Body weights of both ginger-administered groups increased compared to the control. Colon length increased to 7.6, and 8.0 cm in the G and BG groups, respectively, whereas that of control was 5.7 cm. Histological colon injury induced by DSS-induced colitis was reduced (P<0.05). In serum and DSS-treated colon tissues, mRNA expression levels of IFN-${\gamma}$, IL-6, TNF-${\alpha}$, and IL-12 of the Beopje ginger-treated group were significantly suppressed compared to those of the ginger-treated groups. Expression levels of iNOS and COX-2 of the Beopje ginger-treated group were significantly reduced compared to those of the ginger-treated groups (P<0.05), and BG showed stronger anti-inflammatory effects on colitis. These results indicated that ginger exerted anti-inflammatory effects on DSS-induced colitis in mice, and its effects could be increased through Beopje.

First Report of Soybean Dwarf Virus on Soybean(Glycine max) in Korea (콩(Glycine max)에서 콩위축바이러스(Soybean dwarf virus)의 최초 발생보고)

  • Kim, Sang-Mok;Lee, Jae-Bong;Lee, Yeong-Hoon;Choi, Se-Hoon;Choi, Hong-Soo;Park, Jin-Woo;Lee, Jun-Seong;Lee, Gwan-Seok;Moon, Jung-Kyung;Moon, Jae-Sun;Lee, Key-Woon;Lee, Su-Heon
    • Research in Plant Disease
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    • v.12 no.3
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    • pp.213-220
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    • 2006
  • In year 2003, a soybean(Glycine max) sample showing severe dwarfing symptom was collected from a farmers' field in Cheongsong in Korea. The results from the diagnosis of the sample by RT-PCR revealed that it was infected by Soybean dwarf virus(SbDV), SbDV-L81. This study could be the first report of the occurrence of the virus in Korea. To further characterize the virus, the partial nucleotide sequence of the genomic RNA of SbDV-L81 was determined by RT-PCR using species-specific primers. The sequences were analyzed and subsequently compared to previously characterized strains of SbDV based on the pattern of symptom expression and vector specificities. The intergenic region between ORF 2 and 3 and the coding regions of ORF 2, 3 and 4 were relatively similar to those of dwarfing strains(SbDV-DS and DP) rather than those of yellowing strains(SbDV-YS and YP). Likewise, the result from the analysis of 5'-half of the coding region of ORF5 indicated that SbDV-L81 was closely related to strains(SbDV-YP and DP) transmitted by Acyrthosiphon pisum. These data from the natural symptom and the comparisons of five regions of nucleotide sequences of SbDV suggested that SbDV-L81 might be closely related SbDV-DP.

Improvement of colitis preventive effects of Gochujang by addition of Lactobacillus plantarum on C57BL/6 mice (Lactobacillus plantarum 첨가 고추장의 C57BL/6 마우스에서 대장염 예방 증진효과)

  • Park, Eui-Seong;Heo, Ju-Hee;Lim, Yaung-Iee;Ju, Jaehyun;Park, Kun-Young
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1188-1194
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    • 2017
  • Gochujang, a traditional Korean food, is fermented by mixing red pepper powder, various grain, meju and salt. Changes in the kind of ingredients and fermentation method could increase health functionalities. In this study, in vivo anti-colitis effects of gochujang prepared with mixed grains, bamboo salt baked 3 times and meju starters on DSS-induced colitis in C57BL/6 mice were studied. We prepared gochujang prepared with mixed grains (MG), bamboo salt, and Aspergillus oryzae (A) and Baccillus subtilis (B) mixed starters (MG-AB) and gochujang prepared with MG, bamboo salt and A, B and Lactobacillus plantarum (L) mixed starters (MG-ABL). MG-AB and MG-ABL significantly increased body weight and colon length compared to the control (p<0.05). MG-ABL showed significantly decreased interleukin-6 (IL-6) expression in serum compared to the control and MG-AB group (p<0.05). MG-ABL also regulated mRNA and protein levels of pro-apoptotic Bcl-2-associated X protein (Bax) and anti-apoptotic B-cell lymphoma-2 (Bcl-2) in the mice colon tissue (p<0.05). Therefore, MG-ABL exhibited the increased anticolitis effects by inhibiting damage of colon tissue, probably by regulating a pro-inflammatory cytokine of IL-6 and regulated apoptosis related genes. These results indicated that gochujang changed with good ingredients and starters had colitis preventive effects and might be due to active compounds in mixed grain and bamboo salt, and produced by L during the fermentation of gochujang.

Anticancer and Antiviral Activity of Chlorine Dioxide by Its Induction of the Reactive Oxygen Species (이산화염소의 활성산소 생성 유도에 의한 항암 및 항바이러스 활성)

  • Kim, Yonggyun;Kumar, Sunil;Cheon, Wonsu;Eo, Hyunji;Kwon, Hyeok;Jeon, Yongho;Jung, Jinboo;Kim, Wook
    • Journal of Applied Biological Chemistry
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    • v.59 no.1
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    • pp.31-36
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    • 2016
  • Chlorine dioxide has been used for a disinfectant by exhibiting antimicrobial activity and is also potent to kill insect pests infesting stored grains. This study aimed to extend the usefulness of chlorine dioxide with respect to anticancer and antiviral activities. Cytotoxicity of chlorine dioxide was assessed against five different human cancer cell lines. Chlorine dioxide exhibited significant cytotoxicity against two breast cancer cell lines (MCF-7, MDA-MB-231) and three colorectal cancer cell lines (LoVo, HCT-116, SW-480). This cytotoxicity appeared to be associated with the capacity of chlorine dioxide to induce the production of reactive oxygen species (ROS). Compared to control insect cell lines, the cancer cell lines possessed much higher levels of ROS. On the other hand, a treatment of an antioxidant, vitamin E, significantly reduced the cytotoxicity, suggesting that the cytotoxicity was induced by high levels of ROS production. Chlorine dioxide exhibited antiviral activity against different viruses. A baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV), is a dsDNA insect virus and lost its viral activity to form polyhedral viral particles in response to chlorine dioxide. The antiviral activity against AcNPV was dependent on the incubation time with chlorine dioxide. Tobacco mosaic virus is a ssRNA plant virus and was reduced in its population after exposure to chlorine dioxide along with significant decrease of viral symptoms. These results indicate that chlorine dioxide possesses anticancer and antiviral activities probably due to its inducing activity of ROS production.