• Title/Summary/Keyword: abolished protein

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Anti-inflammatory effect of methanol extract from Erigeron Canadensis L. may be involved with upregulation of heme oxygenase-1 expression and suppression of $NF{\kappa}B$ and MAPKs activation in macrophages

  • Sung, Jeehye;Sung, Misun;Kim, Younghwa;Ham, Hyeonmi;Jeong, Heon-Sang;Lee, Junsoo
    • Nutrition Research and Practice
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    • v.8 no.4
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    • pp.352-359
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    • 2014
  • BACKGROUND/OBJECTIVES: In this study, we determined the anti-inflammatory activities and the underlying molecular mechanisms of the methanol extract from Erigeron Canadensis L. (ECM) in LPS-stimulated RAW264.7 macrophage cells. MATERIALS/METHODS: The potential anti-inflammatory properties of ECM were investigated by using RAW264.7 macrophages. We used western blot assays and real time quantitative polymerase chain reaction to detect protein and mRNA expression, respectively. Luciferase assays were performed to determine the transactivity of transcription factors. RESULTS: ECM significantly inhibited inducible nitric oxide synthase (iNOS)-derived NO and cyclooxygenase-2 (COX-2) derived PGE2 production in LPS-stimulated RAW264.7 macrophages. These inhibitory effects of ECM were accompanied by decreases in LPS-induced nuclear translocations and transactivities of $NF{\kappa}B$. Moreover, phosphorylation of mitogen-activated protein kinase (MAPKs) including extracellular signal-related kinase (ERK1/2), p38, and c-jun N-terminal kinase (JNK) was significantly suppressed by ECM in LPS-stimulated RAW264.7 macrophages. Further studies demonstrated that ECM by itself induced heme oxygenase-1 (HO-1) protein expression at the protein levels in dose-dependent manner. However, zinc protoporphyrin (ZnPP), a selective HO-1 inhibitor, abolished the ECM-induced suppression of NO production. CONCLUSIONS: These results suggested that ECM-induced HO-1 expression was partly responsible for the resulting anti-inflammatory effects. These findings suggest that ECM exerts anti-inflammatory actions and help to elucidate the mechanisms underlying the potential therapeutic values of Erigeron Canadensis L.

The Effect of Cold-adaptation on Stress Responses and Identification of a Cold Shock Gene, capA in Bradyrhizobium japonicum (Bradyrhizobium japonicum의 저온 전처리에 의한 환경스트레스 내성 증진에 대한 연구)

  • 유지철;노재상;오은택;소재성
    • Korean Journal of Microbiology
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    • v.38 no.1
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    • pp.45-49
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    • 2002
  • Bradyrhizobium japonicum is a soil bacterium with a unique ability to infect the roots of leguminous plants and establish a nitrogen-fixing symbiosis, which has been used as a microbial manure. In this study, we examined the stress response after pretreatment of cells with cold temperature. When pre-treated with cold temperature ($4^{\circ}C$) for 16 hr, B. japonicum increased the viability in subsequent stress-conditions such as alcohol, $H_2O_2$, heat, and dehydration. For cold adpatation, cultured B. japonicum was exposed to $4^{\circ}C$. Upon subsequent exposure to various conditions, the number of adapted cells pretreated by cold adaptation was 10-1000 fold higher than that of non-adaptated ones. It appeared de novo protein synthesis occurred during adaptation, because a protein synthesis inhibitor, chloramphenicol abolished the increased stress tolerance. By using a degenerate PCR primer set, a csp homolog was amplified from B. japonicum genome and sequenced. The deduced partial amino acid sequence of the putative Csp (Cold shock protein) shares a significant similarity with known Csp proteins of other bacteria.

Effect of Glutamate on the Vestibulo-Solitary Projection after Sodium Nitroprusside-Induced Hypotension in Conscious Rats

  • Li, Li-Wei;Ji, Guang-Shi;Yang, Yan-Zhao;Ameer, Abdul Nasir;Kim, Min Sun;Park, Byung Rim;Jin, Yuan-Zhe
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.3
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    • pp.275-281
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    • 2015
  • Orthostatic hypotension is most common in elderly people, and its prevalence increases with age. Attenuation of the vestibulo-sympathetic reflex (VSR) is commonly associated with orthostatic hypotension. In this study, we investigated the role of glutamate on the vestibulo-solitary projection of the VSR pathway to clarify the pathophysiology of orthostatic hypotension. Blood pressure and expression of both pERK and c-Fos protein were evaluated in the nucleus tractus solitarius (NTS) after microinjection of glutamate into the medial vestibular nucleus (MVN) in conscious rats with sodium nitroprusside (SNP)-induced hypotension that received baroreceptor unloading via sinoaortic denervation (SAD). SNP-induced hypotension increased the expression of both pERK and c-Fos protein in the NTS, which was abolished by pretreatment with glutamate receptor antagonists (MK801 or CNQX) in the MVN. Microinjection of glutamate receptor agonists (NMDA or AMPA) into the MVN increased the expression of both pERK and c-Fos protein in the NTS without causing changes in blood pressure. These results indicate that both NMDA and AMPA receptors play a significant role in the vestibulo-solitary projection of the VSR pathway for maintaining blood pressure, and that glutamatergic transmission in this projection might play a key role in the pathophysiology of orthostatic hypotension.

The p53-p21Cip1/WAF1 Pathway Is Necessary for Cellular Senescence Induced by the Inhibition of Protein Kinase CKII in Human Colon Cancer Cells

  • Kang, Ji-Young;Kim, Jin Joo;Jang, Seok Young;Bae, Young-Seuk
    • Molecules and Cells
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    • v.28 no.5
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    • pp.489-494
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    • 2009
  • We have previously shown that the down-regulation of protein kinase CKII activity is tightly associated with cellular senescence of human fibroblast IMR-90 cells. Here, we examined the roles of p53 and $p21^{Cip1/WAF1}$ in senescence development induced by CKII inhibition using wild-type, isogenic p53-/- and isogenic p21-/- HCT116 human colon cancer cell lines. A senescent marker appeared after staining for senescence-associated ${\beta}$-galactosidase activity in wild-type HCT116 cells treated with CKII inhibitor or $CKII{\alpha}$ siRNA, but this response was almost abolished in p53- or $p21^{Cip1/WAF1}$-null cells. Increased cellular levels of p53 and $p21^{Cip1/WAF1}$ protein occurred with the inhibition of CKII. CKII inhibition upregulated p53 and $p21^{Cip1/WAF1}$ expression at post-transcriptional level and transcription level, respectively. RB phosphorylation significantly decreased in cells treated with CKII inhibitor. Taken together, this study shows that the activation of the $p53-p21^{Cip1/WAF1}$ pathway acts as a major mediator of cellular senescence induced by CKII inhibition.

Curcumin Inhibits Osteoclastogenesis by Decreasing Receptor Activator of Nuclear Factor-κB Ligand (RANKL) in Bone Marrow Stromal Cells

  • Oh, Sora;Kyung, Tae-Wook;Choi, Hye-Seon
    • Molecules and Cells
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    • v.26 no.5
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    • pp.486-489
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    • 2008
  • Curcumin (diferuloylmethane), a pigment derived from turmeric, has anti-oxidant and anti-inflammatory activities. Accumulating evidence points to a biochemical link between increased oxidative stress and reduced bone density. Osteoclast formation was evaluated in co-cultures of bone marrow stromal cells (BMSC) and whole bone marrow cells (BMC). Expression of receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL) was analyzed at the mRNA and protein levels. Exposure to curcumin led to dose-dependent suppression of osteoclastogenesis in the co-culture system, and to reduced expression of RANKL in $IL-1{\alpha}$-stimulated BMSCs. Addition of RANKL abolished the inhibition of osteoclastogenesis by curcumin, whereas the addition of prostaglandin $E_2$ ($PGE_2$) did not. The decreased osteoclastogenesis induced by curcumin may reduce bone loss and be of potential benefit in preventing and/or attenuating osteoporosis.

Improved Cell Viability of Lactobacillus crispatus KLB46 by Stress Adaptation (Lactobacillus crispatus KLB46의 스트레스 전처리시 열 내성 증진효과)

  • Kwak, Dae-Yung;Kang, Chang-Ho;Jeon, HanEul;So, Jae-Seong
    • KSBB Journal
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    • v.29 no.2
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    • pp.81-86
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    • 2014
  • Lactobacilli, the dominant species of microorganisms in the vaginal flora of healthy women, play important roles to prevent bacterial vaginosis and other sexually transmitted diseases. In this study, we carried out studies on stress adaptation prior to various stress treatment. We found that heat or salt adapted KLB46 showed higher cell viability than non adapted upon heat stress at $60^{\circ}C$ for 20 min. When chloramphenicol was added during the adaptation process, heat tolerance was abolished. This result suggested that de novo protein synthesis was essential during adaptation.

Antilisterial Bacteriocin from Lactobacillus rhamnosus CJNU 0519 Presenting a Narrow Antimicrobial Spectrum

  • Jeong, Ye-Jin;Moon, Gi-Seong
    • Food Science of Animal Resources
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    • v.35 no.1
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    • pp.137-142
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    • 2015
  • A lactic acid bacterium presenting antimicrobial activity against a Lactobacillus acidophilus strain used for eradication of acid inhibition was isolated from a natural cheese. The 16S rRNA gene sequence of the isolate best matched with a strain of L. rhamnosus and was designated L. rhamnosus CJNU 0519. The antimicrobial activity of the partially purified bacteriocin of CJNU 0519 was abolished when treated with a protease, indicating the protein nature of the bacteriocin. The partially purified bacteriocin (rhamnocin 519) displayed a narrow antimicrobial activity against L. acidophilus, Listeria monocytogenes, and Staphylococcus aureus among several tested bacterial and yeast strains. Rhamnocin 519 in particular showed strong bactericidal action against L. monocytogenes.

Antimicrobial, Antioxidant and Hemolytic Activity of Water-soluble Extract of Mottled Anemone Urticina crassicornis

  • Lee, Ye Jin;Kim, Chan-Hee;Oh, Hye Young;Go, Hye-Jin;Park, Nam Gyu
    • Fisheries and Aquatic Sciences
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    • v.18 no.4
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    • pp.341-347
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    • 2015
  • We evaluated the biological activities of five water extracts of tissue of the mottled anemone Urticina crassicornis. Most extracts exhibited broad-spectrum antimicrobial activity as determined by ultrasensitive radial diffusion assay (URDA) against gram-positive and -negative bacteria, including a fish pathogen, Aeromonas hydrophila, but no activity against fungi. The activity of the extracts was abolished by tryptic digestion, indicating that protein compounds were responsible for the antimicrobial activity. Furthermore, in a 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity assay, only the visceral tissue extract showed activity. However, no extract had hemolytic activity against human red blood cells. Consequently, this study suggests the water-soluble extract of mottled anemone to be a promising source of proteinaceous antimicrobial compounds that can be utilized for development of novel antibiotics.

Effects of R100 Mutant MerR on Regulation of mer Operon from Shigella flexneri

  • Yoon, Kyung-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.245-249
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    • 1994
  • An amino-terminal 14 amino acids deletion and three site-directed mutations were created to investigate the mechanism of induction and repression of MerR regulatory protein in R100 mer operon from gramnegative Shigella flexneri. The amino-terminal 14 amino acids deletion, Cysl17Ser, and Cys126Ser mutations abolished the inducibility of the mer operon and the Hisl18Ala mutation resulted in the reduction of inducibility (about 9.1 % remaining) in complementation experiment in the presence of $Hg^{2+}$ at subtoxic level ($1\mu M$). The complementation experiment with $Hg^{2+}$ absent showed that Hisl18Ala, Cys126Ser, and wild-type MerR could repress the operon but Cysl17Ser could not, and the amino-terminal deletion mutant could neither induce nor repress the R100 mer operon.

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Effects of α1-adrenoceptor stimulation on Mg2+ release in perfused guinea pig heart (관류 기니픽 심장에서 Mg2+ 유리에 미치는 α1-adrenoceptor 자극효과)

  • Hwang, Sung-chul;Kim, Sang-jin;Kang, Hyung-sub;Lee, Seung-ok;Kang, Chang-won;Kwon, Oh-deog;Kim, Jin-sang
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.327-335
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    • 1996
  • Recently in spite of the interest on the regulation of intracellular $Mg^{2+}$ by neurotransmitters or drugs, the magnesium ion($Mg^{2+}$) regulation by ${\alpha}_1$-adrenoceptor stimulation has not been studied in the heart yet. To elucidate the regulation of ${\alpha}_1$-adrenoceptor stimulation-induced $Mg^{2+}$ release and the effects of ${\alpha}_1$-adrenoceptor stimulation on pathophysiological conditions, in this study we have evaluated the effects of phenylephrine, PMA, $H_7$. staurosporine, verapamil and lidocaine on $Mg^{2+}$ release in perfused guinea pig heart. During preperfusion exogenous $Mg^{2+}$ was added to the medium to give 1.2mM 15min before starting to addition of drugs, and then the infusion of exogenous $Mg^{2+}$ was stopped. $Mg^{2+}$ in the perfusate leaving the heart was measured by atomic absorption spectrophotometry. $Mg^{2+}$ free solution produced an increase in heart rate and phenylephrine elicited $Mg^{2+}$ release from the heart. $Mg^{2+}$ release by phenylephrine was abolished by combined treatment with prazosin. By contrast, cardiac $Mg^{2+}$ uptake induced by a protein kinase C(PKC) activator, PMA was abolished by a selective PKC inhibitor, staurosporine. And the phenylephrine-induced $Mg^{2+}$ release was not affected by the PKC inhibitor, $H_7$. When verapamil or lidocaine was added to perfusing solution, $Mg^{2+}$ release was potentiated by phenylephrine from perfused guinea pig heart. These results suggest that ${\alpha}_1$-adrenoceptor stimulation caused $Mg^{2+}$ release and that PKC is not involved in ${\alpha}_1$-adrenoceptor mediated $Mg^{2+}$ release from perfused guinea pig heart. Under pathophysiological conditions, the $Mg^{2+}$ alteration by ${\alpha}_1$-adrenoceptor stimulation is considerable.

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