• Title/Summary/Keyword: specific RNA

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Anticancer Activity of the Antimicrobial Peptide Scolopendrasin VII Derived from the Centipede, Scolopendra subspinipes mutilans

  • Lee, Joon Ha;Kim, In-Woo;Kim, Sang-Hee;Kim, Mi-Ae;Yun, Eun-Young;Nam, Sung-Hee;Ahn, Mi-Young;Kang, Dongchul;Hwang, Jae Sam
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1275-1280
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    • 2015
  • Previously, we performed de novo RNA sequencing of Scolopendra subspinipes mutilans using high-throughput sequencing technology and identified several antimicrobial peptide candidates. Among them, a cationic antimicrobial peptide, scolopendrasin VII, was selected based on its physicochemical properties, such as length, charge, and isoelectric point. Here, we assessed the anticancer activities of scolopendrasin VII against U937 and Jurkat leukemia cell lines. The results showed that scolopendrasin VII decreased the viability of the leukemia cells in MTS assays. Furthermore, flow cytometric analysis and acridine orange/ethidium bromide staining revealed that scolopendrasin VII induced necrosis in the leukemia cells. Scolopendrasin VII-induced necrosis was mediated by specific interaction with phosphatidylserine, which is enriched in the membrane of cancer cells. Taken together, these data indicated that scolopendrasin VII induced necrotic cell death in leukemia cells, probably through interaction with phosphatidylserine. The results provide a useful anticancer peptide candidate and an efficient strategy for new anticancer peptide development.

Application of Bacteria Isolated from Dok-do for Improving Compressive Strength and Crack Remediation of Cement-sand Mortar (독도산 탄산칼슘형성세균에 의한 모르타르 균열보수와 압축강도 증진)

  • Park, Sung-Jin;Lee, Na-Young;Kim, Wha-Jung;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.216-221
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    • 2010
  • This study shows an application of microbiologically induced carbonate precipitate for strength improvement and crack remediation of cement-sand mortar. Seven calcium carbonate-forming bacteria (CFB) were isolated from Dok-do and partially identified by DNA sequence analysis of the 16s rRNA gene. Crystal aggregates were apparent around the bacterial colonies grown on an agar medium. These strains showed strain specific $CaCO_3$ precipitation on urea-$CaCl_2$ medium. Among 7 isolates, Arthrobacter nicotinovorans KNUC601, Microbacterium resistens KNUC602, Agrobacterium tumefaciens KNUC603, Exiguobacterium acetylicum KNUC604, and Bacillus thuringiensis KNUC606 showed a repairing of artificial forced cracks in cement-sand mortar. Compressive strength of cement-sand mortar consolidated with Stenotrophomonas maltophilia KNUC605 was increased around 14.07% compared with that of negative control.

Inhibition of Escherichia coli O157:H7 Attachment by Interactions Between Lactic Acid Bacteria and Intestinal Epithelial Cells

  • Kim, Young-Hoon;Kim, Sae-Hun;Whang, Kwang-Youn;Kim, Young-Jun;Oh, Se-Jong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1278-1285
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    • 2008
  • The intestinal epithelial cell (IEC) layer of the intestinal tract makes direct contact with a number of microbiota communities, including bacteria known to have deleterious health effects. IECs possess innate protective strategies against pathogenic challenge, which primarily involve the formation of a physicochemical barrier. Intestinal tract mucins are principal components of the mucus layer on epithelial surfaces, and perform a protective function against microbial damage. However, little is currently known regarding the interactions between probiotics/pathogens and epithelial cell mucins. The principal objective of this study was to determine the effects of Lactobacillus on the upregulation of MUC2 mucin and the subsequent inhibition of E. coli O157:H7 attachment to epithelial cells. In the current study, the attachment of E. coli O157:H7 to HT-29 intestinal epithelial cells was inhibited significantly by L. acidophilus A4 and its cell extracts. It is also important to note that the expression of MUC2 mucin was increased as the result of the addition of L. acidophilus A4 cell extracts (10.0 mg/ml), which also induced a significant reduction in the degree to which E. coli O157:H7 attached to epithelial cells. In addition, the mRNA levels of IL-8, IL-1$\beta$, and TNF-$\alpha$ in HT-29 cells were significantly induced by treatment with L. acidophilus A4 extracts. These results indicate that MUC2 mucin and cytokines are important regulatory factors in the immune systems of the gut, and that selected lactobacilli may be able to induce the upregulation of MUC2 mucin and specific cytokines, thereby inhibiting the attachment of E. coli O157:H7.

Changes in Gut Microbial Community of Pig Feces in Response to Different Dietary Animal Protein Media

  • Jeong, Yujeong;Park, Jongbin;Kim, Eun Bae
    • Journal of Microbiology and Biotechnology
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    • v.30 no.9
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    • pp.1321-1334
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    • 2020
  • Beef, pork, chicken and milk are considered representative protein sources in the human diet. Since the digestion of protein is important, the role of intestinal microflora is also important. Despite this, the pure effects of meat and milk intake on the microbiome are yet to be fully elucidated. To evaluate the effect of beef, pork, chicken and milk on intestinal microflora, we observed changes in the microbiome in response to different types of dietary animal proteins in vitro. Feces were collected from five 6-week-old pigs. The suspensions were pooled and inoculated into four different media containing beef, pork, chicken, or skim milk powder in distilled water. Changes in microbial communities were analyzed using 16S rRNA sequencing. The feces alone had the highest microbial alpha diversity. Among the treatment groups, beef showed the highest microbial diversity, followed by pork, chicken, and milk. The three dominant phyla were Proteobacteria, Firmicutes, and Bacteroidetes in all the groups. The most abundant genera in beef, pork, and chicken were Rummeliibacillus, Clostridium, and Phascolarctobacterium, whereas milk was enriched with Streptococcus, Lactobacillus, and Enterococcus. Aerobic bacteria decreased while anaerobic and facultative anaerobic bacteria increased in protein-rich nutrients. Functional gene groups were found to be over-represented in protein-rich nutrients. Our results provide baseline information for understanding the roles of dietary animal proteins in reshaping the gut microbiome. Furthermore, growth-promotion by specific species/genus may be used as a cultivation tool for uncultured gut microorganisms.

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.

Characterization of Expressed Sequence Tags (ESTs) Generated from the Bombyx mandarina Whole Larvae and Molecular Cloning of Serine Protease Homologue Gene

  • Hwang, Jae Sam;Yun, Eun Young;Goo, Tae Won;Kim, Iksoo;Choi, Kwang Ho;Seong, Su Il;Kim, Keun Young;Lee, Sang Mong;Kang, Seok Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.2
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    • pp.167-171
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    • 2004
  • We constructed an oligo-d(T) primed directional cDNA library from the Bombyx mandarina whole larvae. In an effort to isolate genes expressed in the B. mandarina, 227 expressed sequence tags (ESTs) were generated by single-pass sequencing from the cDNA library. Sequence analysis showed that 107 clones (47.1%) were classified into known genes and 120 clones (52.9%) were novel transcripts, which are unknown for their function. Of the 107 known genes, the most abundant gene was found to be actin and followed by serine protease in the expression profile. Among these clones, a serine protease homolog (BmSP) which is a class of proteolytic enzymes isolated. Full-length sequence of the BmSP cDNA clone was 922 bp in length and has an open reading frame of 276 amino acids. The conserved histidine, aspatic acid and serine residues forming the catalytic center as well as cysteine residues contributing to three disulphide bonds also were found in Bmsp gene. mRNA expression analysis revealed a high and specific expression of the gene only in midgut tissue, suggesting that BmSP gene is closely associated with the expression of digestive enzyme.

Sochungyong-tang, Tradititional Korean Medicine, Suppresses Th2 Lineage Development (소청룡탕(小靑龍湯)이 분화된 Th1 cell 및 Th2 cell cytokine profile에 미치는 영향)

  • Jeong, Hyuk-Joon;Hong, Moo-Chang;Shin, Min-Kyu;Bae, Hyun-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.2
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    • pp.380-388
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    • 2005
  • In this study, the immunological effect of a traditional Korea herbal medicine, Sochungyong-tang (SCRT) that has been widely used for the treatment of various immunological disorders including allergic asthma in Korea, was examined in vitro. In our previous study demonstrated that SCRT decreases the expression of IL-4 mRNA, that plays pivotal role in Th2 cell development, while increases $IFN-{\gamma}{\tilde{a}}expression$, which is one of the key cytokines for Th1 lineage development in Th0 condition. That study strongly implies that SCRT can correct Th2 dominant condition directly affecting to the CD4+ T cell development. Present study designated to further evaluate the SCRT on helper T cell development by monitoring Th1/Th2 specific cytokine secretion patterns in artificially induced Th1 or Th2 polarized condition. The results demonstrated that Th2 cells were dramatically under-populated in Th2 driven condition with SCRT treatment, while Th1 cells were not altered in Th1 skewed condition. Furthermore, under Th2-skewed conditions the levels of and IL-4 were considerably decreased with SCRT treatment. However, the expression of GATA-3, a transcription factor that plays pivotal role in Th2 lineage programming, was not changed with SCRT, suggesting that the suppression of Th2 cell development by SCRT was not mediated by GATA-3. Present study implies that the effect on CD4+ T cell may be the one of key pharmacological effect point for treating IgE medicated allergic asthma by SCRT. These results also suggest that SCRT might be desirable agent for the correction of Th2 dominant pathological disorders.

Anti-allergic Effects of Jagamcho-tang in Ovalbumin-induced Atopic Dermatitis Mouse Model (Ovalbumin으로 유발된 아토피피부염 마우스 모델에서 자감초탕(炙甘草湯)의 항알레르기 효과)

  • Noh, Hyeon-Min;Park, Sung-gu;Kim, Wonjeong;Jo, Eun-Hee;Ki, Hyeon-Hui;Kim, Dae-Ki;Park, Min-Cheol
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.31 no.5
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    • pp.277-283
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    • 2017
  • Activated T helper 2 (Th2) immune function is hallmark of various allergic diseases. We investigated the anti-allergic effect of Jagamcho-tang extract(JE) on ovalbumin(OVA)-induced atopic dermatitis mice model and OVA-stimulated splenocytes isolated from the mice. Mice were intraperitoneally injected OVA/alum solution 2 times at interval of 14 days, followed by oral administration of JE for 7 days. After administration, mice were subcutaneously injected with OVA in ear. JE treatment reduced ear swelling and infiltration of inflammatory cells in ear. Serum levels of interleukin(IL)-4 and immunoglobulins, such as total-IgE and OVA-specific IgE, were decreased in JE treated group. Furthermore, JE treatment decreased OVA-induced Th2-associated cytokines like IL-4, IL-5 and IL-13 mRNA levels in splenocytes. In conclusion, JE reduced allergic immune response via IgE production and Th2 response in OVA-sensitized mice, suggesting that JE could be useful prescription for allergic diseases including atopic dermatitis.

Effects of Resveratrol and trans-3,5,4'-Trimethoxystilbene on Glutamate-Induced Cytotoxicity, Heme Oxygenase-1, and Sirtuin 1 in HT22 Neuronal Cells

  • Kim, Dae-Won;Kim, Young-Mi;Kang, Sung-Don;Han, Young-Min;Pae, Hyun-Ock
    • Biomolecules & Therapeutics
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    • v.20 no.3
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    • pp.306-312
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    • 2012
  • Resveratrol (trans-3,5,4'-trihydroxystilbene) has received considerable attention recently for the potential neuroprotective effects in neurodegenerative disorders where heme oxygenase-1 (HO-1) and sirtuin 1 (SIRT1) represent promising therapeutic targets. Resveratrol has been known to increase HO-1 expression and SIRT1 activity. In this study, the effects of resveratrol and trans-3,5,4'-trimethoxystilbene (TMS), a resveratrol derivative, on cytotoxicity caused by glutamate-induced oxidative stress, HO-1 expression, and SIRT1 activation have been investigated by using murine hippocampal HT22 cells, which have been widely used as an in vitro model for investigating glutamate-induced neurotoxicity. Resveratrol protected HT22 neuronal cells from glutamate-induced cytotoxicity and increased HO-1 expression as well as SIRT1 activity in a concentration-dependent manner. Cytoprotection afforded by resveratrol was partially reversed by the specific inhibition of HO-1 expression by HO-1 small interfering RNA and the nonspecific blockage of HO-1 activity by tin protoporphyrin IX, but not by SIRT1 inhibitors. Surprisingly, TMS, a resveratrol derivative with methoxyl groups in lieu of the hydroxyl groups, and trans-stilbene, a non-hydroxylated analog, failed to protect HT22 cells from glutamate-induced cytotoxicity and to increase HO-1 expression and SIRT1 activity. Taken together, our findings suggest that the cytoprotective effect of resveratrol was at least in part associated with HO-1 expression but not with SIRT1 activation and, importantly, that the presence of hydroxyl groups on the benzene rings of resveratrol appears to be necessary for cytoprotection against glutamate-induced oxidative stress, HO-1 expression, and SIRT1 activation in HT22 neuronal cells.

L1 Cell Adhesion Molecule Promotes Migration and Invasion via JNK Activation in Extrahepatic Cholangiocarcinoma Cells with Activating KRAS Mutation

  • Kim, Haejung;Hwang, Haein;Lee, Hansoo;Hong, Hyo Jeong
    • Molecules and Cells
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    • v.40 no.5
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    • pp.363-370
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    • 2017
  • Extrahepatic cholangiocarcinoma (ECC), a malignant tumor of biliary origin, has a poor prognosis with limited treatment options. The KRAS oncogene is the most commonly mutated gene in ECC and one of the factors that predicts a poor prognosis and low survival rate. L1 cell adhesion molecule (L1CAM) is expressed in ECC cells and acts as an independent poor prognostic factor in predicting patient survival. In this study we investigate the functional significance of L1CAM in ECC cells with activating KRAS mutation. We selected an ECC cell line, EGI-1, with activating KRAS mutation, and then confirmed its expression of L1CAM by RT-PCR, western blot analysis, and flow cytometry. The suppression of L1CAM expression (using a specific lentivirus-delivered shRNA) significantly decreased the migratory and invasive properties of EGI-1 cells, without altering their proliferation or survival. Analyses of signaling effectors in L1CAM-depleted and control EGI-1 cells indicated that L1CAM suppression decreased the levels of both phosphorylated MKK4 and total MKK4, together with c-Jun N-terminal kinase (JNK) phosphorylation. Further, exposure to a JNK inhibitor (SP600125) decreased migration and invasion of EGI-1 cells. These results suggest that L1CAM promotes cellular migration and invasion via the induction of MKK4 expression, leading to JNK activation. Our study is the first to demonstrate a functional role for L1CAM in ECC carrying the activating KRAS mutation. Given that KRAS is the most commonly mutated oncogene in ECC, L1CAM may serve as an attractive therapeutic target for ECC cells with activating KRAS mutation.