• Title/Summary/Keyword: NHE8 protein

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Up-regulation of NHE8 by somatostatin ameliorates the diarrhea symptom in infectious colitis mice model

  • Lei, Xuelian;Cai, Lin;Li, Xiao;Xu, Hua;Geng, Chong;Wang, Chunhui
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.3
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    • pp.269-275
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    • 2018
  • $Na^+/H^+$ exchangers (NHEs) have been shown to be involved in regulating cell volume and maintaining fluid and electrolyte homeostasis. Pooled evidences have suggested that loss of $Na^+/H^+$ exchanger isoform 8 (NHE8) impairs intestinal mucosa. Whether NHE8 participates in the pathology of infectious colitis is still unknown. Our previous study demonstrated that somatostatin (SST) could stimulate the expression of intestinal NHE8 so as to facilitate $Na^+$ absorption under normal condition. This study further explored whether NHE8 participates in the pathological processes of infectious colitis and the effects of SST on intestinal NHE8 expression in the setting of infectious colitis. Our data showed that NHE8 expression was reduced in Citrobacter rodentium (CR) infected mice. Up-regulation of NHE8 improved diarrhea symptom and mucosal damage induced by CR. In vitro, a similar observation was also seen in Enteropathogenic E. coli (EPEC) infected Caco-2 cells. Seglitide, a SST receptor (SSTR) 2 agonist, partly reversed the inhibiting action of EPEC on NHE8 expression, but SSTR5 agonist (L-817,818) had no effect on the expression of NHE8. Moreover, SST blocked the phosphorylation of p38 in EPEC-infected Caco-2 cells. Taken together, these results suggest that enhancement of intestinal NHE8 expression by SST could ameliorate the symptoms of mice with infectious colitis.

Melanogenesis Inhibition Effects of Nemopilema nomurai Hydrolized Extracts (노무라입깃해파리 가수분해 추출물의 멜라닌합성 저해효과)

  • Lee, Ha Youn;Kim, Young Hoon;Um, Sun Young;Jung, Uk Sun;Chang, Moon Sik;Lee, Nam Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.4
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    • pp.341-347
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    • 2014
  • This study was aimed to investigate skin whitening effects of Nomura's jellyfish (Nemopilema nomurai) hydrolyzed extracts (NHE). The extracts were prepared through the hydrolysis of N. nomurai using commercial proteolytic enzymes such as Protamex, Alcalase, Flavourzyme and Neutrase with optimum pHs (pH 5-8) at $55^{\circ}C$. Their whitening activities were examined from the inhibition of melanin synthesis using B16-F1 cell lines. Among the examined samples, Neutrase-treated extract (N-NHE) showed the most significant inhibition effect on melanin synthesis by 89.9% at a concentration of $100{\mu}g/mL$. This sample decreased the expression of tyrosinase and TRP-1 (tyrosinase related protein-1) proteins as well. These results suggested the potential application of NHE as whitening ingredients in cosmetic preparation.

Cloning and Activity Analysis of the FosB Promoter Region from Human Genomic DNA (사람 핵DNA로부터 FosB 유전자 프로모터 클로닝 및 활성도 분석)

  • Na, Han-Heom;Kang, Yoonsung;Kim, Keun-Cheol
    • Journal of Life Science
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    • v.27 no.8
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    • pp.857-863
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    • 2017
  • The FBJ murine osteosarcoma viral oncogene homolog B (FosB) gene is located at chromosome 19, and encodes 43 Kda protein. Functionally, the FosB gene is important for differentiation, development, and pathogenesis. Furthermore, the FosB gene is suggested as possible biomarker for tracing disease prognosis. In this study, we constructed plasmid containing a FosB promoter region and evaluate its promoter activity. We analyzed the putative promoter region in FosB genomic DNA using bioinformatics program, and we found important regulatory elements in 1 Kb upstream from transcription start site (TSS). Therefore, we performed polymerase chain reaction (PCR) amplification on region from-1,555 upstream to +73 of the FosB genomic DNA, and PCR product was inserted into TA vector to create the $TA-1^{st}FosBp$ plasmid. We then prepared the primer sets, which contain a restriction enzyme site for Kpn1 and Nhe1, in order to reinsert into the TA vector to prepare $TA-2^{nd}FosBp$ plasmid. It was finally subcloned into pGL3-luc vector after enzyme cutting. To evaluate whether the cloned plasmid is useful in cell based experiment, we performed luciferase assay with pGL3-FosBp-luctransfection. FosB promoter activity was increased compared to empty vector, and this activity was significantly increased by treatment of doxorubicin and taxol. We obtained consistent data on regulation of FosB gene expression after anticancer drug treatment using Western blot analysis. The results suggest that promoter cloning of the human FosB gene is very useful for studying gene expression and analyzing biomarkers.

Sequence Analysis of a Cryptic Plasmid pKW2124 from Weissella cibaria KLC140 and Construction of a Surface Display Vector

  • Kim, Soo Young;Oh, Chang Geun;Lee, Young Joo;Choi, Kyu Ha;Shin, Doo Sik;Lee, Si Kyung;Park, Kab Joo;Shin, Hakdong;Park, Myeong Soo;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.545-554
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    • 2013
  • Plasmid isolation of kimchi-derived Weissella cibaria KLC140 revealed six different plasmids. The smallest plasmid, pKW2124, was DNA sequenced and characterized, showing 2,126 bp with a GC content of 36.39% and five putative open reading frames (ORFs). In silico analysis of these ORFs showed ORF1 encodes a putative replication protein similar to rolling circular replication proteins from other lactic acid bacteria. However, a single-stranded intermediate was not detected when S1 nuclease was treated, suggesting it may follow theta replication. Interestingly, the replication initiation site of this plasmid is 100% identical to other plasmids from lactic acid bacteria, suggesting it may function for replication initiation. To construct a surface layer expression vector, pTSLGFP, slpA encoding the surface layer protein from Lactobacillus acidophilus was PCR amplified and fused with the gfp gene, forming a SLGFP fused gene. The plasmid pKW2124 was cloned into the XbaI site of pUC19, forming an Weissella-E. coli shuttle vector pKUW22. NheI-linearized pTSLGFP was ligated into pKUWCAT containing pKUW22 and the chloramphenicol acetyltransferase gene from pEK104, resulting in an 8.6 kb pKWCSLGFP surface layer expression vector. After transformation of this vector into W. cibaria KLC140, a GFP fluorescence signal was detected on the surface of the transformant, substantiating production of SLGFP fused protein and its secretion. This is the first report for construction of a Weissella surface layer expression vector, which may be useful for surface layer production of beneficial proteins in Weissella.

Physicochemical characteristics of garlic (Allium sativum L.) shoots from different areas in Namhae (남해지역 마늘종의 이화학적 특성)

  • Yoon, Hwan-Sik;Kang, Min-Jung;Hwang, Cho-Rong;Sim, Hye-Jin;Kim, Gyeong-Min;Shin, Jung-Hye
    • Food Science and Preservation
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    • v.21 no.3
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    • pp.321-327
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    • 2014
  • This paper introduces the physicochemical characteristics of garlic shoots investigated in the 8 areas of Namhae, Gohyeon-myeon (GHM), Nam-myeon (NM), Samdong-myeon (SDM), Namhae-eup (NHE), Seo-myeon (SM), Seolcheon-myeon (SCM), Idong-myeon (IDM) and Changsoen-myeon (CSM). The moisture was $80.15{\pm}0.37{\sim}83.87{\pm}0.15g/100g$. The crude protein and lipid were $1.59{\pm}0.02{\sim}2.25{\pm}0.09g/100g$ and $0.35{\pm}0.01{\sim}0.48{\pm}0.03g/100g$, respectively. Ash was $0.65{\pm}0.03{\sim}0.71{\pm}0.03g/100g$. In addition, the total levels of phenol and flavonoid were searched as $33.02{\pm}1.78{\sim}58.41{\pm}1.08mg/100g$ and $11.60{\pm}3.36{\sim}23.95{\pm}2.04mg/100g$, respectively. The garlic shoots in IDM, where the highest total chlorophyll level detected as $170.58{\pm}1.04{\mu}g/g$ and in CSM, where the lowest total chlorophyll level investigated as $116.65{\pm}0.00{\mu}g/g$, were unveiled. The mineral elements (S, Ca, Fe, K, Mg, Na, Mn and Zn) were analyzed by ICP-OES, and K was the most highly contained mineral with $7257.0{\pm}87.9{\sim}10070.0{\pm}9.7mg/kg$. Significant feature of garlic shoots in IDM and NHE was that the total amounts of mineral were relatively higher than other regions as 21,143.5 mg/kg and 20,581.2 mg/kg, respectively. The cultivating regions were revealed as the one of important matters with the level of allicin ($78.03{\pm}6.15{\sim}203.43{\pm}18.47mg/kg$). In particular, the level of allicin of garlic raised in SDM ($203.43{\pm}18.47mg/kg$) was approximately 2.2~2.6 times higher than those in CSM ($92.86{\pm}8.70mg/kg$) and SCM ($78.03{\pm}6.15mg/kg$). The sulfur compounds content of diallyl trisulde was remarkably higher, especially the garlic shoots in IDM with $830.62{\pm}14.69{\sim}1281.10{\pm}27.71mg/kg$.