• Title/Summary/Keyword: cell infection

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Inhibition of the Replication of Hepatitis C Virus Replicon with Nuclease-Resistant RNA Aptamers

  • Shin, Kyung-Sook;Lim, Jong-Hoon;Kim, Jung-Hye;Myung, Hee-Joon;Lee, Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1634-1639
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    • 2006
  • Hepatitis C virus (HCV)-encoded nonstructural protein 5B (NS5B) possesses RNA-dependent RNA polymerase activity, which is considered essential for viral proliferation. Thus, HCV NS5B is a good therapeutic target protein for the development of anti-HCV agents. In this study, we isolated two different kinds of nuclease-resistant RNA aptamers with 2'-fluoro pyrimidines against the HCV NS5B from a combinatorial RNA library with 40 nucleotide random sequences, using SELEX technology. The isolated RNA aptamers were observed to specifically and avidly bind the HCV NS5B with an apparent $K_d$ of 5 nM and 18 nM, respectively, in contrast with the original RNA library that hardly bound the target protein. Moreover, these aptamers could partially inhibit RNA synthesis of the HCV subgenomic replicon when transfected into Huh-7 hepatoma cell lines. These results suggest that the RNA aptamers selected in vitro could be useful not only as therapeutic agents of HCV infection but also as a powerful tool for the study of the HCV RNA-dependent RNA polymerase mechanism.

Anti-inflammatory effect of new calcium hydroxide paste containing silicon-substituted hydroxyapatite in lipopolysaccharide-stimulated macrophages

  • Roh, Ji-Yeon;Kim, Ki-Rim
    • Journal of Korean society of Dental Hygiene
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    • v.18 no.4
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    • pp.423-432
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    • 2018
  • Objectives: Calcium hydroxide, a root canal temporary sealer, has long been used and it has anti-bacterial and anti-inflammatory activity. To investigate the properties of a newly developed calcium hydroxide paste comprising silicon-substituted hydroxyapatite (Si-HA), we examined the anti-inflammatory activity of the new calcium hydroxide paste in RAW 264.7 macrophages stimulated by lipopolysaccharide (LPS), which causes infection of the root canal. Methods: The test materials, including Calcipex II as control group and the newly developed TRC paste, were extracted from cell culture media and then diluted for experiment. In LPS-stimulated RAW 264.7 cells, the cytotoxicity and nitric oxide (NO) production of test materials were measured by MTT assay and Griess reagents, respectively. Also, the expression of the inducible nitric oxide synthase (iNOS) was assessed by western blotting. Results: The IC50 values of Calcipex II and TRC paste were $17.6mg/m{\ell}$ and $13.5mg/m{\ell}$, respectively. The level of NO, increased by LPS, was dose-dependently inhibited more by TRC paste than Calcipex II treatment. In addition, iNOS expression was decreased by 71% and 92% at concentrations of $2mg/m{\ell}$ and $20mg/m{\ell}$ of TRC paste, respectively. Conclusions: We demonstrated that the Si-HA calcium hydroxide paste has a slightly improved anti-inflammatory property and further studies are needed before clinical recommendations are proposed.

The Effect of Various Partition Extracts of Radix Codonopsis Lanceolatae on the Melanogenesis (더덕 분획별 抽出液이 멜라닌 生成에 미치는 影響)

  • Oh, Han-Cheol;Hong, Chul-Hee;Lee, Soo-Hyeong;Hwang, Chung-Yeon;Kim, Nam-Kwon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.17 no.2
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    • pp.59-71
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    • 2004
  • Melanin pigmentation has an essential role in protecting human skin against the damaging effects of ultra violet radiation, infection, environmental factor, etc. However abnormal melanin pigmentation can directly cause a number of congenital and acquired pigmentary skin disease. This study conducted to evaluate the effects of various partition extracts of Codonopsis lanceolata on the melanogenesis in the B 16 melanoma cells. The cells were treated for 3 days with various partition extracts of Codonopsis lanceolata. While treatment with the BuOH partition extract of Codonopsis lanceolata increased significantly the tyrosinase activity and melanin content, the n-Hexane or ethyl acetate partition extract dose-dependently suppressed them compared with untreated control. But ethyl acetate partition significantly decreased the cell viability. The H2O partiton extract at various concentrations did not exhibit any significant change on the melanogenesis. These results indicate that treatment with the n-Hexane partition extract inhibits the melanogenesis in B16 melanoma cells, and raise the possibility that this extract may be effective in the abnormal melanin pigmentation as well as the whitening agent for the skin.

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Altered Sulfate Metabolism of Arabidopsis Caused by Beet Severe Curly Top Virus Infection

  • Lee, Hong-Gun;Park, Sung-Hee;Kim, Dong-Giun;Lee, Taek-Kyun;Yum, Seung-Shic;Auh, Chung-Kyoon;Lee, Suk-Chan
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.355-360
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    • 2005
  • Sulfur, an important component of plants, is regulated by a variety of stresses in sulfate assimilation and metabolism. Increase has been observed in the expression of O-acetylserine(thiol)lyase (OASTL) through two-dimensional electrophoresis with the shoot tips of Arabidopsis infected by beet severe curly top geminivirus (BSCTV). With the three- to six-fold increases in the transcript expression of OASTL, serine acetyltransferase (SAT) and $\gamma$-glutmylcysteine synthetase (GSH) were induced over the mock-inoculated organization in each organization through real-time RT-PCR analysis. The expression of those genes might affect the accumulation of anthocyanin in symptomatic tissues and the induction of abnormal callus-like structures formed by additional cell divisions as typical disease symptoms of BSCTV-infected Arabidopsis. This is the first report to describe the collaborative induction of OASTL, SAT, and GSH in virus-infected plants. The changed expressions of OASTL, SAT, and GSH in Arabidopsis infected with BSCTV raises new aspects regarding the biological function of symptomatic tissues related to sulfate metabolism.

Purification and biochemical characterization of two novel antigens from Leishmania major promastigotes

  • Zeinali, Majid;Ardestani, Sussan K.;Kariminia, Amina
    • Parasites, Hosts and Diseases
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    • v.45 no.4
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    • pp.287-293
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    • 2007
  • The identification and characterization of antigens that elicit human T cell responses is an important step toward understanding of Leishmania major infection and ultimately in the development of a vaccine. Micropreparative SDS-PAGE followed by electro transfer to a PVDF membrane and elution of proteins from the PVDF, was used to separate 2 novel proteins from L. major promastigotes, which can induce antibodies of the IgG2a isotype in mice and also are recognized by antisera of recovered human cutaneous leishmaniasis subjects. Fractionation of the crude extract of L. major revealed that all detectable proteins of interest were present within the soluble Leishmania antigens (SLA). Quantitation of these proteins showed that their expression in promastigotes is relatively very low. Considering the molecular weight, immunoreactivity, chromatographic and electrophoretic behavior in reducing and non-reducing conditions, these proteins are probably 2 isoforms of a single protein. A digest of these proteins was resolved on Tricine-SDS-PAGE and immunoreactive fragments were identified by human sera. Two immunoreactive fragments (36.4 and 34.8 kDa) were only generated by endoproteinase Glu-C treatment. These immunoreactive fragments or their parent molecules may be ideal candidates for incorporation in a cocktail vaccine against cutaneous leishmaniasis.

3D-printed titanium implant with pre-mounted dental implants for mandible reconstruction: a case report

  • Park, Jung-Hyun;Odkhuu, Michidgerel;Cho, Sura;Li, Jingwen;Park, Bo-Young;Kim, Jin-Woo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.42
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    • pp.28.1-28.4
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    • 2020
  • Background: This clinical case presented a novel method of segmental mandible reconstruction using 3D-printed titanium implant with pre-mounted dental implants that was planned to rehabilitate occlusion. Case presentation: A 53-year-old male who suffered osteoradionecrosis due to the radiation after squamous cell carcinoma resection. The 3D-printed titanium implant with pre-mounted dental implant fixtures was simulated and fabricated with selective laser melting method. The implant was successfully inserted, and the discontinuous mandible defect was rehabilitated without postoperative infection or foreign body reaction during follow-ups, until a year. Conclusions: The 3D-printed titanium implant would be the one of the suitable treatment modalities for mandible reconstruction considering all the aspect of mandibular functions.

Characterization of rock bream (Oplegnathus fasciatus) fin cells and its susceptibility to different genotypes of megalocytiviruses

  • Jeong, Ye Jin;Kim, Young Chul;Min, Joon Gyu;Jeong, Min A;Kim, Kwang Il
    • Journal of fish pathology
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    • v.34 no.2
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    • pp.149-159
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    • 2021
  • Genus Megalocytivirus cause red sea bream iridoviral disease (RSIVD) and scale drop disease (SDD). Based on the phylogeny of the major capsid protein (MCP) and adenosine triphosphatase (ATPase) genes, megalocytiviruses except for SDD virus (SDDV) could be three different genotypes, red sea bream iridovirus (RSIV), infectious spleen and kidney necrosis (ISKNV), and turbot reddish body iridovirus (TRBIV). In this study, primary cells derived from the caudal fin of rock bream (Oplegnathus fasciatus) grew at 25℃ in Leibovitz's medium supplemented with 10% (v/v) fetal bovine serum and primocin (100 ㎍/mL). Rock bream fin (RBF) cells exhibited susceptibility to infections by different genotypes of megalocytiviruses (RSIV, ISKNV and TRBIV) with the appearance of cytopathic effects with an increase in the viral genome copy number. Furthermore, compared to grunt fin (GF) cells, even though 10 times lower number of RSIV genome copies were inoculated in RBF cells, viral genome copy number produced on RBF cells were 44 times higher than that of GF cells at 7 d post-inoculation. As the isolated RBF cells are sensitive to different genotypes of megalocytiviruses (RSIV, ISKNV and TRBIV), they can be used for future studies regarding in vitro viral infection and subsequent diagnosis.

Multi-Epitope Fusion Protein Eg mefAg-1 as a Serodiagnostic Candidate for Cystic Echinococcosis in Sheep

  • Tianli, Liu;Xifeng, Wang;Zhenzhong, Tian;Lixia, Wang;Xingxing, Zhang;Jun, Qiao;Qingling, Meng;Shasha, Gong;Ying, Chen;Xuepeng, Cai
    • Parasites, Hosts and Diseases
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    • v.57 no.1
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    • pp.61-67
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    • 2019
  • Cystic echinococcosis (CE) in sheep is a hazardous zoonotic parasitic disease that is caused by Echinococcus granulosus (Eg). At present, serological test is an important diagnostic method for Eg infection in domestic animals. Here, a fusion protein Eg mefAg-1 harboring 8 dominant B-cell epitopes of Eg such as antigen B, tetraspanin 1, tetraspanin 6, reticulon and Eg95 was produced in E. coli and evaluated for CE in sheep by indirect ELISA. Eg mefAg-1 showed in ELISA a high sensitivity (93.41%) and specificity (99.31%), with a coincidence rate of 97.02%. Overall, it is suggested that the Eg mefAg-1 could be a potential antigen candidate for CE serodiagnosis in sheep.

Mini-review: oomycete RXLR genes as effector-triggered immunity

  • Arif, Saima;Jang, Hyun A;Kim, Mi-Reu;Oh, Sang-Keun
    • Korean Journal of Agricultural Science
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    • v.45 no.4
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    • pp.561-573
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    • 2018
  • Oomycetes are known to secrete a vast arsenal of effectors that modulate the host defense system as well as facilitate establishing a parasitic infection in plants. In recent years, tremendous progress has been made in the field of effectromics based on studies of oomycetes, especially the cytoplasmic family of RXLR effectors. Yet, the biology of the RXLR effector family is still poorly understood. There has been a consensus regarding the structure of the RXLR motif in the mycologist community. However, the function of the RXLR motif is still unclear. First, different models have suggested that the role of the RXLR motif is either in translocation to a target destination inside a host cell or in the cleavage of itself followed by secretion. Second, recent studies have suggested different functional models for the RXLR motif. According to a widely accepted model, the RXLR motif is directly involved in the translocation of effectors to target sites. In contrast, a new study has proposed that the RXLR motif is involved in secretion rather than translocation. Thus, this review is an attempt to summarize the recent advances made in the functional analysis of the N-terminal domain of RXLR effectors.

Improved Cell Viability and Anti-Candida Activity of Probiotic Lactobacillus salivarius MG242 by Heat Adaptation (Lactobacillus salivarius MG242의 열 전처리시 생존율 증진 및 항 캔디다 효과)

  • Kang, Chang-Ho;Kim, YongGyeong;Shin, YuJin;Paek, Nam-Soo;So, Jae-Seong
    • Journal of Dairy Science and Biotechnology
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    • v.37 no.1
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    • pp.49-56
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    • 2019
  • Vulvovaginal candidiasis is a major urogenital infection in women. Lactobacilli are important in maintaining vaginal health. In the present study, the effect of heat adaptation at $47{\sim}52^{\circ}C$ prior to heat stress at $60^{\circ}C$ in improving the viability of Lactobacillus salivarius MG242 was examined. L. salivarius MG242 has antifungal effects against Candida albicans. Heat-adapted cells had a higher survival rate than non-adapted cells during the subsequent heat stress. When chloramphenicol was added during the adaptation process, heat tolerance was abolished, suggesting the involvement of de novo protein synthesis with the heat adaptation of L. salivarius MG242 strain. Exopolysaccharide quantification and scanning election microscopy did not reveal any appreciable changes during heat adaptation. The antifungal activity of L. salivarius MG242 against C. albicans was maintained during the heat adaptation. These results suggest that heat adaptation can be applied for the development of probiotic products using L. salivarius MG242 to improve its stress tolerance during processing.