• Title/Summary/Keyword: STAT2

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Effect of Soyangin-Hyeongbangpaedok-san on Anti-CD3 Stimulated Mouse T Cells In Vivo and In Vitro (소양인 형방패독산(荊防敗毒散)의 마우스 T 세포 활성에 대한 실험적 연구)

  • Kim, Joong-Bae;Kang, Hee;Ahn, Kwang-Seok;Shim, Bum-Sang;Kim, Sung-Hoon;Choi, Seung-Hoon;Ahn, Kyoo-Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.3
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    • pp.554-561
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    • 2009
  • Soyangin-Hyeongbangpaedok-san(SHBPDS) is a herbal formula used for the common cold or upper respiratory illness. In order to investigate the effect of SHBPDS, mice were orally administered with SHBPDS alcohol extract for 7 days followed by intravenous anti-CD3 injection. In addition, splenocytes and CD4 T cells were cultured with SHBPDS in response to anti-CD3 in vitro and cytokines and transcription factors were evaluated. In vivo treatment with SHBPDS significantly augmented the expressions of the percentage of CD4 T cells and CD 69, an indicator of early T cell activation. Serum levels of IL-4 were significantly increased but those of IFN-${\gamma}$ and IL-2 did not reach statistical significance. The expressions of IFN-${\gamma}$ and T-bet mRNA were significantly downregulated in SHBPDS treated mice while those of IL-4 and C-Maf were significantly upregulated. In vitro stimulation of splenocytes and CD4 T cells by SHBPDS resulted in a reduction in IFN-${\gamma}$ secretion and STAT4 activity. The IL-4 releases from both cells were slightly reduced, but STAT6 activity was rather increased. In conclusion, SHBPDS exerted an inhibition in the expression of IFN-${\gamma}$, T-bet and STAT4 while IL-4, C-Maf and STAT6 were increased. Further studies are required to examine its pharmacological effects using more appropriate animal experiments.

Role of IFNLR1 gene in PRRSV infection of PAM cells

  • Qin, Ming;Chen, Wei;Li, Zhixin;Wang, Lixue;Ma, Lixia;Geng, Jinhong;Zhang, Yu;Zhao, Jing;Zeng, Yongqing
    • Journal of Veterinary Science
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    • v.22 no.3
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    • pp.39.18-39.18
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    • 2021
  • Background: Interferon lambda receptor 1 (IFNLR1) is a type II cytokine receptor that clings to interleukins IL-28A, IL29B, and IL-29 referred to as type III IFNs (IFN-λs). IFN-λs act through the JAK-STAT signaling pathway to exert antiviral effects related to preventing and curing an infection. Although the immune function of IFN-λs in virus invasion has been described, the molecular mechanism of IFNLR1 in that process is unclear. Objectives: The purpose of this study was to elucidate the role of IFNLR1 in the pathogenesis and treatment of porcine reproductive and respiratory syndrome virus (PRRSV). Methods: The effects of IFNLR1 on the proliferation of porcine alveolar macrophages (PAMs) during PRRSV infection were investigated using interference and overexpression methods. Results: In this study, the expressions of the IFNLR1 gene in the liver, large intestine, small intestine, kidney, and lung tissues of Dapulian pigs were significantly higher than those in Landrace pigs. It was determined that porcine IFNLR1 overexpression suppresses PRRSV replication. The qRT-PCR results revealed that overexpression of IFNLR1 upregulated antiviral and IFN-stimulated genes. IFNLR1 overexpression inhibits the proliferation of PAMs and upregulation of p-STAT1. By contrast, knockdown of IFNLR1 expression promotes PAMs proliferation. The G0/G1 phase proportion in IFNLR1-overexpressing cells increased, and the opposite change was observed in IFNLR1-underexpressing cells. After inhibition of the JAK/STAT signaling pathway, the G2/M phase proportion in the IFNLR1-overexpressing cells showed a significant increasing trend. In conclusion, overexpression of IFNLR1 induces activation of the JAK/STAT pathway, thereby inhibiting the proliferation of PAMs infected with PRRSV. Conclusion: Expression of the IFNLR1 gene has an important regulatory role in PRRSV-infected PAMs, indicating it has potential as a molecular target in developing a new strategy for the treatment of PRRSV.

Candida magnoliae에 의한 erythritol 생산을 위한 유가식 공정의 개발

  • Park, Chang-Yeol;Seo, Jin-Ho;Yu, Yeon-U
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.53-56
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    • 2000
  • Two-stage fed-batch culture was peformed to improve the volumetric productivity of erythritol. In the growth phase dissolved oxygen was maintained to 20% and the feed medium was automatically supplied to the fermenter by pH-stat mode. The cell yield was 0.76 g-cell/g-glucose. In two-stage fed-batch culture, 41% of total erythritol conversion yield with 187 g/L of erythritol concentration and 2.79 g/L-h of maximum erythritol Productivity were obtained when 400 g/L of glucose was directly added in the form of non-sterile powder at production phase. The erythritol productivity increased in parallel with cell mass. The metabolic shift in the biosynthetic pathway of erythritol was caused by dissolved oxygen concentration. The production of gluconic acid was observed when the dissolved oxygen in the medium was maintained over 40% during the production phase, whereas the dissolved oxygen concentration lower than 40% caused the production of citric acid. But the butyric acid was produced independently with dissolved oxygen concentration in the medium. The production of organic acids such as gluconic acid, citric acid, and butyric acid was decreased by addition of mineral salts.

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Methylated Alteration of SHP1 Complements Mutation of JAK2 Tyrosine Kinase in Patients with Myeloproliferative Neoplasm

  • Yang, Jun-Jun;Chen, Hui;Zheng, Xiao-Qun;Li, Hai-Ying;Wu, Jian-Bo;Tang, Li-Yuan;Gao, Shen-Meng
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2219-2225
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    • 2015
  • SHP1 negatively regulates the Janus kinase 2/signal transducer and activator of transcription (JAK2/STAT) signaling pathway, which is constitutively activated in myeloproliferative neoplasms (MPNs) and leukemia. Promoter hypermethylation resulting in epigenetic inactivation of SHP1 has been reported in myelomas, leukemias and other cancers. However, whether SHP1 hypermethylation occurs in MPNs, especially in Chinese patients, has remained unclear. Here, we report that aberrant hypermethylation of SHP1 was observed in several leukemic cell lines and bone marrow mononuclear cells from MPN patients. About 51 of 118 (43.2%) MPN patients including 23 of 50 (46%) polycythaemia vera patients, 20 of 50 (40%) essential thrombocythaemia and 8 of 18 (44.4%) idiopathic myelofibrosis showed hypermethylation by methylation-specific polymerase chain reaction. However, SHP1 methylation was not measured in 20 healthy volunteers. Hypermethylation of SHP1 was found in MPN patients with both positive (34/81, 42%) and negative (17/37, 45.9%) JAK2V617F mutation. The levels of SHP1 mRNA were significantly lower in hypermethylated samples than unmethylated samples, suggesting SHP1 may be epigenetically inactivated in MPN patients. Furthermore, treatment with 5-aza-2'-deoxycytidine (AZA) in K562 cells showing hypermethylation of SHP1 led to progressive demethylation of SHP1, with consequently increased reexpression of SHP1. Meanwhile, phosphorylated JAK2 and STAT3 were progressively reduced. Finally, AZA increased the expression of SHP1 in primary MPN cells with hypermethylation of SHP1. Therefore, our data suggest that epigenetic inactivation of SHP1 contributes to the constitutive activation of JAK2/STAT signaling. Restoration of SHP1 expression by AZA may contribute to clinical treatment for MPN patients.

Effect of Environmental pH on End Products, Fermentation Balances and Bioenergetic As-pects of Lactobacillus bulg-aricus in a Glucose-limited “pH Stat” Continuous Culture.

  • Rhee, Sang-Ki;Pack, Moo-Young
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1979.10a
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    • pp.246.1-246
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    • 1979
  • A glucose-limited “pH-stat” continuous culture study of Lactobacillus bulgaricus NLS-4 in an anaerobic condition showed the marked effects of environmental pH on end products, fermentation blances and bioenergetic aspects of the organism. Lactic acid was the major end product of fermentation with minor products, such as acetic acid, formic acid and ethanol throughout the pH range tested. In acidic conditions below pH 6.5, a typi-cal pattern of homofermentation was revealed whereas in alkaline conditions, the metabolic pattern was changed from homofermentation to heterofermentation and led to acquire much energy. This metabolic change was likely due to the pH-dependent lactate dehydrogenase activity. Molar growth yields (Yglc=35.5-44.4) and YATP, $18.5\pm2.5$ in average which was 80% higher than the value ever postulated seemed to be accounted for less requirement of maintenance energy of the organism in the culture conditions.

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Extracellular Overproduction of $\beta$-Cyclodextrin Glucanotransferase in a Recombinant E. coli Using Secretive Expression System

  • Lee, Kwang-Woo;Shin, Hyun-Dong;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.753-759
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    • 2002
  • $\beta$-Cyclodextrin glucanotransferase ($\beta$-CGTase) was overproduced extracellularly using recombinant E. coli by transforming the plasmid pECGT harboring a secretive signal peptide. The $\beta$-CGTase gene of alkalophilic Bacillus firmus var alkalophilus was inserted into the high expression vector pET20b(+) containing a secretive pelB signal peptide, and then transformed into E. coli BL2l(DE3)pLysS. The optimum culture conditions fer the overproduction of $\beta$-CGTase were determined to be TB medium containing 0.5% (w/v) soluble starch at post-induction temperature of $25^{\circ}C$. A significant amount of $\beta$-CGTase, up to 5.83 U/ml, which was nine times higher than that in the parent strain B. firmus var. alkalophilus, was overproduced in the extracellular compartment. A pH-stat fed-batch cultivation of the recombinant E. coli was also performed to achieve the secretive overproduction of $\beta$-CGTase at a high cell density, resulting in production of up to 21.6 U/ml of $\beta$-CGTase.

Comparative Analysis of Growth-Phase-Dependent Gene Expression in Virulent and Avirulent Streptococcus pneumoniae Using a High-Density DNA Microarray

  • Ko, Kwan Soo;Park, Sulhee;Oh, Won Sup;Suh, Ji-Yoeun;Oh, TaeJeong;Ahn, Sungwhan;Chun, Jongsik;Song, Jae-Hoon
    • Molecules and Cells
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    • v.21 no.1
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    • pp.82-88
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    • 2006
  • The global pattern of growth-dependent gene expression in Streptococcus pneumoniae strains was evaluated using a high-density DNA microarray. Total RNAs obtained from an avirulent S. pneumoniae strain R6 and a virulent strain AMC96-6 were used to compare the expression patterns at seven time points (2.5, 3.5, 4.5, 5.5, 6.0, 6.5, and 8.0 h). The expression profile of strain R6 changed between log and stationary growth (the Log-Stat switch). There were clear differences between the growth-dependent gene expression profiles of the virulent and avirulent pneumococcal strains in 367 of 1,112 genes. Transcripts of genes associated with bacterial competence and capsular polysaccharide formation, as well as clpP and cbpA, were higher in the virulent strain. Our data suggest that late log or early stationary phase may be the most virulent phase of S. pneumoniae.

Root Bark extract of Morus alba L. Suppressed the Migration and Invasion of HCT116 Human Colorectal Carcinoma Cells (HCT116 인체 대장암 세포주에서 상백피 추출물에 의한 전이 억제 효과)

  • Park, Shin-Hyung;Park, Hyun-Ji
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.35 no.5
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    • pp.177-184
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    • 2021
  • The root bark of Morus alba L. (MA) used in traditional oriental medicine for the treatment of pulmonary diseases exerts various pharmacological activities including anticancer effects. In the current study, we investigated the effects of MA on the migration and invasion of colorectal carcinoma cells. Results from a transwell assay showed that the methylene chloride extract of MA (MEMA) suppressed the migration and invasion of HCT116 human colorectal carcinoma cells in a concentration-dependent manner. MEMA reduced both mRNA and protein levels of matrix metalloproteinase (MMP)-9, but did not suppress the expression of MMP-2 in HCT116 cells. As a molecular mechanism, MEMA inhibited the phosphorylation of mitogen-activated protein kinases (MAPKs), including ERK, JNK and p38, in a dose-dependent manner. In addition, MEMA dephosphorylated both Src and signal transducer and activator of transcription 3 (STAT3) in HCT116 cells. Taken together, we demonstrate that MEMA suppressed the migration and invasion capacity of HCT116 human colorectal cancer cells by downregulation of MMP-9 and inactivation of both MAPKs and Src/STAT3 signaling pathway.

Based on morphology and molecular data, Palisada rigida comb. nov. and Laurencia decussata comb. et stat. nov. (Rhodophyta, Rhodomelaceae) are proposed

  • Metti, Yola
    • ALGAE
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    • v.37 no.1
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    • pp.15-32
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    • 2022
  • Inspecting herbaria collections of Laurencia rigida highlighted frequent misidentifications between L. rigida and L. heteroclada f. decussata, two poorly studied taxa from Australia. Recent collections of DNA material, including from topotype material, allowed for re-examination of these two taxa using molecular techniques. Detailed morphological and molecular analyses based on two markers (rbcL and COI-5P) strongly supported these two taxa as being distinct from each other and requiring nomenclatural changes. Comprehensive morphological analyses highlighted features useful for accurate identifications. Interestingly, L. rigida was found to belong to the genus Palisada with evidence from both the morphology and molecular data. Therefore, this study proposed recognizing L. rigida as Palisada rigida comb. nov. Molecular data for L. heteroclada f. decussata on the other hand supported its separation from L. heteroclada, with too great a molecular distance to be considered a variety. Morphological characters that best separated P. rigida from L. decussata included seven characters; number of pericentral cells per vegetative axial segment, the presence of secondary pit connections, the presence of lenticular thickenings, tetrasporangia alignment, the presence of corps en cerise, holdfast morphology, and overall plant shape. Morphologically, L. heteroclada f. decussata was also separated from L. heteroclada, particularly by the following characteristics; ultimate branchlets morphologies, lower order branch lengths, primary axis and holdfast morphologies. Therefore, it was proposed that L. heteroclada f. decussata is recognized at a species level as L. decussata comb. et stat. nov.

Ethyl Acetate Fraction of Adenophora triphylla var. japonica Inhibits Migration of Lewis Lung Carcinoma Cells by Suppressing Macrophage Polarization toward an M2 Phenotype

  • Park, Shin-Hyung
    • Journal of Pharmacopuncture
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    • v.22 no.4
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    • pp.253-259
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    • 2019
  • Objectives: It is reported that tumor-associated macrophages (TAMs) contribute to cancer progression by promoting tumor growth and metastasis. The purpose of this study is to investigate the effect of different fractions of Adenophora triphylla var. japonica (AT) on the polarization of macrophages into the M2 phenotype, a major phenotype of TAMs. Methods: We isolated hexane, ethyl acetate, and butanol fractions from crude ethanol extract of AT. The cytotoxicity of AT in RAW264.7 cells was examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RAW264.7 cells were polarized into the M2 phenotype by treatment with interleukin (IL)-4 and IL-13. The expression of M2 macrophage marker genes was detected by reverse transcription polymerase chain reaction (RT-PCR). The phosphorylation level of signal transducer and activator of transcription 6 (STAT6) was investigated by western blot analysis. The migration of Lewis lung carcinoma (LLC) cells was examined by transwell migration assay using conditioned media (CM) collected from RAW264.7 cells as a chemoattractant. Results: Among various fractions of AT, the ethyl acetate fraction of AT (EAT) showed the most significant suppressive effect on the mRNA expression of M2 macrophage markers, including arginase-1, interleukin (IL)-10 and mannose receptor C type 1 (MRC-1), up-regulated by treatment of IL-4 and IL-13. In addition, EAT suppressed the phosphorylation of STAT6, a critical regulator of IL-4 and IL-13-induced M2 macrophage polarization. Finally, the increased migration of Lewis lung carcinoma (LLC) cells by CM from M2-polarized RAW264.7 cells was reduced by CM from RAW264.7 cells co-treated with EAT and M2 polarization inducers. Conclusion: We demonstrated that EAT attenuated cancer cell migration through suppression of macrophage polarization toward the M2 phenotype. Additional preclinical or clinical researches are needed to evaluate its regulatory effects on macrophage polarization and anti-cancer activities.