• Title/Summary/Keyword: PLC activity

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Saucerneol F, a New Lignan Isolated from Saururus chinensis, Attenuates Degranulation via Phospholipase Cγ1 Inhibition and Eicosanoid Generation by Suppressing MAP Kinases in Mast Cells

  • Lu, Yue;Son, Jong-Keun;Chang, Hyeun Wook
    • Biomolecules & Therapeutics
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    • v.20 no.6
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    • pp.526-531
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    • 2012
  • During our on-going studies to identify bioactive compounds in medicinal herbs, we found that saucerneol F (SF), a naturally occurring sesquilignan isolated from Saururus chinensis (S. chinensis), showed in vitro anti-inflammatory activity. In this study, we examined the effects of SF on the generation of 5-lipoxygenase (5-LO) dependent leukotriene $C_4$ ($LTC_4$), cyclooxygenase-2 (COX-2) dependent prostaglandin $D_2$ ($PGD_2$), and on phospholipase $C{\gamma}1$ ($PLC{\gamma}1$)-mediated degranulation in SCF-induced mouse bone marrow-derived mast cells (BMMCs). SF inhibited eicosanoid ($PGD_2$ and $LTC_4$) generation and degranulation dose-dependently. To identify the molecular mechanisms underlying the inhibition of eicosanoid generation and degranulation by SF, we examined the effects of SF on the phosphorylation of $PLC{\gamma}1$, intracellular $Ca^{2+}$ influx, the translocation of cytosolic phospholipase $A_2$ ($cPLA_2$) and 5-LO, and on the phosphorylation of MAP kinases (MAPKs). SF was found to reduce intracellular $Ca^{2+}$ influx by inhibiting $PLC{\gamma}1$ phosphorylation and suppressing the nuclear translocations of $cPLA_2$ and 5-LO via the phosphorylations of MAPKs, including extracellular signal-regulated protein kinase-1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38. Taken together, these results suggest that SF may be useful for regulating mast cell-mediated inflammatory responses by inhibiting degranulation and eicosanoid generation.

Antioxidant and Nicotine Degradation Effects of Medicinal Herbs

  • Lee, Ho-Jae;Hur, Sung-Ho;Shin, Eui-Cheol;Lee, Yang-Bong
    • Preventive Nutrition and Food Science
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    • v.8 no.3
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    • pp.289-293
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    • 2003
  • Antioxidant activity and nicotine degradation activity (NDA) of 21 medicinal herbs were determined by using a 1,1-diphenol-2-picrylhydrazyl (DPPH) method and a PLC/PRF5 human liver cell line method, respectively, to develop an anti-smoking aid. The highest and lowest antioxidant activities represented by $IC_{50}$/ value were 30 $\mu$g/mL of Eugenia caryophyllus and 3,270 $\mu$g/mL of Panax ginseng C. A. Meyer, respectively. Antioxidant activity of Eugenia caryophyllus was equal to 38.0$\pm$1.2 mg VCEAC(vitamin C equivalent antioxidant capacity)/ g herb. The highest and lowest NDA values were 1.81 of Astrgalus membranaceus Bunge and 1.01 of Raphani seed and Lespedeza tomentosa Sieb, respectively. Eleven medicinal herbs with high antioxidant activity and/or NDA were selected to make an herbal tea. The herbal tea had high antioxidant activity (50 $\mu$g/mL $IC_{50}$/ and 22.4$\pm$1.4 mgVCEAC/g) and NDA (1.243). The medicinal herb tea could help smokers quitting smoking by degrading and exhausting nicotine accumulated in body and removing reactive oxygen species.

Exploration of Virulence Markers and Genes of Listeria monocytogenes Isolated from Animal Products (축산물유래 Listeria monocytogenes의 virulence marker 및 gene 조사)

  • Yi, Chul-Hyeon;Song, Hyeon-Ho;Kim, Mi-Ryung;Kang, Ho-Jo;Son, Won-Geun
    • Journal of Food Hygiene and Safety
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    • v.23 no.3
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    • pp.248-256
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    • 2008
  • To investigate the epidemiological characteristics of 68 Listeria monocytogenes isolates, including 11 reference strains and 57 isolates from imported US beef, domestic meats(beef, pork, chicken meat), raw milk, and milk plants. L. monocytogenes was to evaluate the production of virulence proteins, such as hemolysin(LLO) and lecithinase(LCP), the adsorption of Congo red(CRA), and to detect virulence genes using the polymerase chain reaction(PCR). In the study of virulence protein production, 68(100%), 62(91.2%), and 54(79.4%) of the 68 L. monocytogenes strains were positive for LLO production, the LCP test, and the CRA test, respectively, while strains of other species, such as L. innocua, L. gray, L. murrayi, and L. welshimeri, were not. There were no significant differences between L. monocytogenes serotypes and the ability to produce LLO or LCP. L. monocytogenesstrains had very high hemolytic titers(2 to 16 fold), while the other Listeria species, other than L. ivanovii and L. seeligeri, did not. The hemolysin activities of L. monocytogenes, L. ivanovii, and L. seeligeri usually exceeded 1.0 HU/mg, while those of other Listeria spp. were less than 0.04 HU/mg. In the PCR assay, all of the L. monocytogenes strains contained the hlyA, plcA, plcB, inlA, and inlB virulence genes and produced a product of the expected size. In the PCR of the actA gene, the expected 385-bp product was seen in 39(57.4%) L. monocytogenesstrains, while an unexpected 268-bp product was seen in 29(42.6%) strains. Most L. monocytogenes strains isolated from Hanwoo beef produced the 385-bp actA gene product, while strains of imported US beef usually produced the 268-bp actA gene product. By contrast, no virulence gene products were amplified in the other Listeria spp.

A Study on the Reflective Practice Experience of a Home Economics Teacher Professional Learning Community Operator (가정과 교사학습공동체 운영자의 성찰적 실천 경험에 관한 실행연구)

  • Lee, Gyeong Suk;Yoo, Taemyung
    • Journal of Korean Home Economics Education Association
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    • v.29 no.2
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    • pp.1-22
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    • 2017
  • The purpose of this research is to share the experience of reflective practice of a Home Economics teacher professional learning community(PLC) operator and to share better suggestions through reflection on PLC operation. All conversation in the 18 sessions of the PLC totally from May 31st 2013 to May 19th 2014 was recorded and transcribed. All materials of PLC activity were qualitatively analysed. Its themes were grounded on its coding and categorization scheme. Findings and conclusions of this study are as followings. The experience of learning the PLC for a year has found that it is the power to keep the PLC alive: courage and anxiety to face anxiety and anxiety about new challenges, the importance of theme selection, a teacher with a reckless challenge, shares of becoming a leader. Through reflection, I learned that concerns require attention to other teachers and the need for a 'New Round' for ongoing meetings. I, as a operator, did not fully consider participant's different interest as well as a program for a new round due to the lack of management experience. It led to the low participations in latter sessions of PLC. Suggestions are put as solutions to improve these problems; (1) grouping participants by school levels when collecting participants, (2) setting operation period and program applying participants' opinions, and (3) operating short-term PLC with certain themes to immerse reaching goal and satisfaction in short time.

Bioluminescent assay of Phospholipase C Using A Luminescent Marine Mutant Bacterium Vibrio harveyi M-17

  • Cho, Ki-Woong;Mo, Sang-Jun;Lee, Hyi-Seung;Park, Jung-Rae;Jongheon Shin
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.150-155
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    • 2000
  • A bioluminescent assay method for detecting the activity of phospholipase C(PLC; phosphatidyl choline cholinephosphohydrolase, EC 3.1.4.3) was developed using bioluminescent marine bacteria. Phospholipase C from Bacillus cereus and sn-1,2- dimyristoyl glycerol was further hydrolyzed with lipase from Candida ecylidracea. The hydrolyzed myristic acid was quantified using a dark mutant of Vibrio harveyi (designated as M-17). The in vivo light intensity of which was stimulated specifically up to one thousand fold in the presence of myristic acid. The rates of the hdrolysis of the DMPC substrate by the phospholipase measured by the luminescence method were linear with time and the were estalished to detect as little as 0.1 mUnit of phospholipase C and 5 nM of myristic acid production.

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Shengmaisan Regulates Pacemaker Potentials in Interstitial Cells of Cajal in Mice

  • Kim, Byung Joo
    • Journal of Pharmacopuncture
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    • v.16 no.4
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    • pp.36-42
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    • 2013
  • Objectives: Shengmaisan (SMS) is a traditional Chinese medicine prescription widely used for the treatment of diverse organs in Korea. The interstitial cells of Cajal (ICCs) are pacemaker cells that play an important role in the generation of coordinated gastrointestinal (GI) motility. We have aimed to investigate the effects of SMS in the ICCs in the mouse small intestine. Methods: To dissociate the ICCs, we used enzymatic digestions from the small intestine in a mouse. After that, the ICCs were identified immunologically by using the anti-c-kit antibody. In the ICCs, the electrophysiological whole-cell patch-clamp configuration was used to record pacemaker potentials in the cultured ICCs. Results: The ICCs generated pacemaker potentials in the mouse small intestine. SMS produced membrane depolarization with concentration-dependent manners in the current clamp mode. Pretreatment with a $Ca^{2+}$ free solution and thapsigargin, a $Ca^{2+}$-ATPase inhibitor in the endoplasmic reticulum, stopped the generation of the pacemaker potentials. In the case of $Ca^{2+}$-free solutions, SMS induced membrane depolarizations. However, when thapsigargin in a bath solution was applied, the membrane depolarization was not produced by SMS. The membrane depolarizations produced by SMS were inhibited by U-73122, an active phospholipase C (PLC) inhibitors. Furthermore, chelerythrine and calphostin C, a protein kinase C (PKC) inhibitors had no effects on SMS-induced membrane depolarizations. Conclusions: These results suggest that SMS might affect GI motility by modulating the pacemaker activity through an internal $Ca^{2+}$- and PLC-dependent and PKC-independent pathway in the ICCs.

Isolation of Anguillosporal, a Potent Inhibitor of Phospholipase C from Fungi Isolate No. 51005 (곰팡이 분리주 No. 51005로부터 포스포리파제 C 저해물질 앙길로스포랄의 분리)

  • Oh, Won-Keun;Lee, Hyun-Sun;Ahn, Soon-Cheol;Ko, Hack-Ryong;Kim, Bo-Yeon;Min, Tae-Ick;Ahn, Jong-Seog
    • YAKHAK HOEJI
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    • v.41 no.4
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    • pp.433-438
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    • 1997
  • MT 51005 as a potent inhibitor of phospholipase C(PLC) was purified from the culture broth of a fungal strain No. 51005 isolated from soil. It was identified as a benzaldehyde d erivative, anguillosporal. by the physico-chemical properties and spectroscopic data. Anguillosporal showed the inhibitory activity against purified PLC with an $IC_{50}\;of\;13{\mu}g/ml$. And it also inhibited the formation of inositol phosphates($IP_t$) in platelet-derived growth factor(PDGF)-stimulated $NIH3T3{\gamma}1$ cells with an $IC_{50}\;of\;0.8{\mu}g/ml$.

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Chemotactic Effect of Leukotactin-1/CCL15 on Human Neutrophils

  • Lee Ji-Sook;Yang Eun-Ju;Ryang Yong-Suk;Kim In-Sik
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.145-151
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    • 2006
  • Leukotactin-l (Lkn-l )/CCL15 has been known as a potent chemoattractant of leukocytes. However, the precise function of Lkn-l in human neutrophils has not been explained well. In the present study, we investigated the contribution of Lkn-1 in chemotactic activity of human neutrophils. Both CCR1 and CCR3 mRNA expressions are strongly expressed in human neutrophils but CCR2 protein expression was uniquely detected on the cell surface. Lkn-l binding to CCR1 and CCR3 induced chemotactic activity of neutrophils. Chemotactic index of Lkn-l was comparable to that of IL-8. $MIP-1{\alpha}/CCL3$ binding to CCR1 and CCR5 has no effect on neutrophil migration. Cell migration, in response to Lkn-l, was blocked by pertussis toxin (Ptx), a $G_o/G_i$ protein inhibitor, and U73122, a phospholipase C(PLC) inhibitor but not by protein kinase C inhibitor such as rottlerin, and Ro-31-8425. Taken together, our results demonstrate that Lkn-l transduces the chemotaxis signal through $G_o/G_i$ protein and PLC. This finding provides the molecular mechanism by which Lkn-l may contribute to neutrophil movement into the site of inflammation.

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Regulation properties of phospholipase C$\delta$ cloned from Misgurnus mizolepis

  • Kim, Na-Young;Ahn, Sang-Jung;Jeon, Soo-Jin;Seo, Jung-Soo;Kim, Moo-Sang;Lee, Sang-Hwan;Je, Ju-Eun;Sung, Ji-Hea;Lee, Hyung-Ho;Lee, June-Woo;Chung, Joon-Ki
    • Journal of fish pathology
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    • v.20 no.2
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    • pp.119-127
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    • 2007
  • Phosphoinositide-specific phospholipase Cδ (PLCδ) plays an important role in many cellular responses and is involved in the production of second messenger. The present study was conducted to characterize the catalytic and regulatory properties of the PLCδ of Misgurnus mizolepis (ML-PLCδ). The ML-PLCδ gene was cloned and expressed under according to the method of the previous report (Kim et al., 2004), and its recombinant protein was purified by successive chromatography using Ni2+-NTA affinity column. The recombinant ML-PLCδ showed a concentration-dependent PLC activity to phosphatidylinositol 4,5-bisphosphate (PIP2) or phosphatidylinositol (PI). Its activity was absolutely Ca2+-dependence, which was similar to mammalian PLCδ isozymes. The Ca2+ concentration yielding maximal activation of ML-PLCδ was 100 μM. However, the activity was decreased interestingly by a polyamine, such as spermine and spermidine. In vitro assay using cholate-micelle cell, ML-PLCδ activity was inhibited in dose-dependent manner by sphinogosine but increased by phosphocholine . In the lipid-binding assay, ML-PLCδ was strongly bound to LPA, PI(3)P, PI(4)P, PI(5)P, PI(3,5)P2, PI(4,5)P2, PI(3,4,5)P3 and PA, but it showed the low affinity to S1P, PI(3,4)P2 and PS. Taken together our results, it is suggested that the general catalytic and regulatory properties of ML-PLCδ are similar with those of mammalian PLCδ1 isozymes, but the N-terminal extended piscine phospholipase Cδ1 (ML-PLCδ) might reflect some distinctions in regulatory properties and inositol-lipid binding specificity between piscine ML-PLCδ and mammalian PLCδ isozymes.

The Survey of Domestic and Foreign for On-Line Monitoring System at Low Voltage Facilities (저전압 전기설비의 온라인 감시시스템 구축을 위한 국내외 실태조사)

  • Kim, Young-Seok;Shong, Kil-Mok;Kim, Sun-Gu
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.21 no.5
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    • pp.75-81
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    • 2007
  • In this paper, the survey and the analysis of domestic and foreign for on-line monitoring system at low voltage facilities example for value added service about electrical safety were executed. According to survey, in Japan, they are executing the on-line electrical facilities monitoring in based on IT basic law for the information of society construction and not only electrical safety but also value added service such as alarm for earthquake, security are being done. In USA and Europe using power line communication(PLC), the on-line observation and building district management service were being realized. Observing the domestic case, electrical safety related with service such as distribution panel product incorporating load management and leakage current measurement was being executed in some companies. From the survey, it is necessary to improve on the law and the system for market activity in domestic.