• Title/Summary/Keyword: $cPLA_2$

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Cromakalim Blocks Membrane Phosphoinositide Activated Signals in the Guinea Pig Lung Mast Cells Stimulated with Antigen-Antibody Reactions

  • Ro, Jai-Youl;Kim, Ji-Young;Kim, Kyung-Hwan
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.2
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    • pp.251-260
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    • 1998
  • Cromakalim (BRL 34915), known as an airway smooth muscle relaxant, inhibited the releases of mediators in the antigen-induced mast cell activation. It has been suggested that cromakalim, in part, inhibited mediator releases by inhibiting the initial increase of 1,2-diacylglycerol (DAG) produced by the activation of the other phospholipase system which is different from phosphatidylcholine-phospholipase D pathway. The aim of this study is to further examine the inhibitory mechanism of cromakalim on the mediator release in the mast cell activation. Guinea pig lung mast cells were purified by using enzyme digestion and percoll density gradient. In purified mast cells prelabeled with $[^3H]PIP_2$, phospholipase C (PLC) activity was assessed by the production of $[^3H]$insitol phosphates. Protein kinase C (PKC) activity was assessed by measuring the protein phosphorylated from mast cells prelabeled with $[{\gamma}-32P]ATP$, and Phospholipase $A_2\;(PLA_2)$ activity by measuring the lyso-phosphatidylcholine produced from mast cell prelabeled with 1-palmitoyl-2-arachidonyl $phosphatidyl-[^{14}C]choline$. Histamine was assayed by fluorometric analyzer, and leukotrienes by radioimmunoassay. The PLC activity was increased by activation of the passively sensitized mast cells. This increased PLC activity was decreased by cromakalim pretreatment. The PKC activity increased by the activation of the passively sensitized mast cells was decreased by calphostin C, staurosporine and cromakalim, respectively. The $PLA_2$ activity was increased in the activated mast cells. The pretreatment of cromakalim did not significantly decrease $PLA_2$ activity. These data show that cromakalim inhibits histamine release by continuously inhibiting signal transduction processes which is mediated via PLC pathway during mast cell activation, but that cromakalim does not affect $PLA_2$ activity related to leukotriene release.

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Effect of the Inhibition of PLA2 and PAF on the Neutrophilic Respiratory Burst and Apoptosis (호중구의 Respiratory Burst에 미치는 PLA2 및 PAF와 영향 : In vitro에서의 호중구의 산소기 생성 및 Apoptosis에 관한 연구)

  • Lee, Young-Man;Kim, Sang-Gyung;Park, Yoon-Yub
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.6
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    • pp.887-897
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    • 2000
  • Background : Since the exact pathogenesis of sepsis-induced ARDS has not been elucidated, the mechanisms of enhanced neutrophilic respiratory burst were probed in endotoxin primed neutrophils associated with the roles of phospholipase A2(PLA2), platelet activating factor(PAF) and apoptosis. Methods : In isolated fresh human neutrophils, effects of the inhibition of PLA2 and PAF on the apoptosis were examined by the method of Annexin-FITC/dual PIflow cytometry. The roles of PLA2 and PAF on the neutrophilic respiratory burst were also examined by measuring oxidant generation in cytochrome-c reduction assay. Activities of the PLA2 and lysoPAF acetyltransferase (lysoPAF AT) of the neutrophils were determined to understand the effect of endotoxin on these enzymatic activities which may be related to the neutrophilic respiratory burst and apoptosis. In addition, the role roles of PLA2 and PAF in neutrophilic adhesion to bovine endothelial cells were examined in vitro by neutrophil adhesion assay. To investigate the effect of oxidants on pulmonary surfactant, cytochemical ultrastructural microscopy was performed. To inhibit PLA2 and PAF, non-specific PLA2 inhibitor mepacrine (100 nM) and WEB 2086 (100 nM) or ketotifen fumarate (10 ${\mu}g$/ml) were used respectively in all in vitro experimental sets. WEB 2086 is PAF receptor antagonist, and ketotifen fumarate is a lyso PAF AT inhibitor. Results: The mapacrine treatment, provided and the endotoxin (ETX) treatment, resulted in increased apoptosis of neutrophils (p<0.001) while treatments of WEB 2086 and ketotifen did not. The inhibition of PLA2 and PAF decreased (p<0.001) production of oxidants from PMA-stimulated neutrophils. While endotoxin increased the PLA2 activity of neutrophils (p<0.01), mepacrine supressed (p<0.001) the activity, provided after treatment of ETX. The lyso PAF actyltransferase activity (lyso PAF AT) increased (p<0.01) after treatment of ETX. In contrast, mepacrine, WEB 2086 and ketotifen showed a tendency of decreasing the activity after treatment of ETX. The treatment of ETX incresed (p<0.001) neutrophil adhesion to endothelial cells, which was reversed by inhibition of PLA2 and PAF (p<0.01). The binding of oxidants to pu1monary surfactant was identified histologically. Conclusions : The enhanced neutrophilic respiratory burst by ETX plays a pivotal role in the pathogenesis of ARDS in terms of oxidayive oxidative stress. Increased production of oxidants from neutrophils is mediated by the activations of PLA2 and lyso PAF AT.

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Moxifloxacin Alleviates Oleic Acid-provoked Neutrophilic Respiratory Burst in the Rat Lung through the Inhibition of Cytosolic Phospholipase $A_2$ (Moxifloxacin의 Cytosolic Phospholipase $A_2$ 억제효과가 흰 쥐 호중구의 Respiratory Burst에 미치는 영향)

  • Lee, Young-Man
    • Tuberculosis and Respiratory Diseases
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    • v.69 no.4
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    • pp.256-264
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    • 2010
  • Background: According to the notion of the immunoregulatory functions of moxifloxacin (MFX), the effect of MFX on the neutrophilic respiratory burst in conjunction with the expression of cytosolic phospholipase $A_2$ ($cPLA_2$) was investigated. Methods: The effects and possible mechanisms of MFX on neutrophilic respiratory burst in oleic acid (OA)-induced acutely injured rats lung and OA-stimulated, isolated murine neutrophils were probed, associated with the expression of cytosolic phospholipase $A_2$ in vivo and in vitro. Results: In the OA-induced acutely-injured lungs, neutrophils were accumulated, which was attenuated by MFX. The parameters denoting a neutrophilic respiratory burst, such as nitro blue tetrazolium reaction, cytochrome-c reduction, neutrophil aggregation, $H_2O_2$ production in neutrophils revealed increased neutrophilic respiratory burst by OA, and MFX decreased all of these parameters. In addition, the enhanced expression of $cPLA_2$ in the lung and isolated murine neutrophils by OA were decreased by MFX. Conclusion: MFX suppresses the OA-induced neutrophilic respiratory burst by the suppression of $cPLA_2$ in neutrophils.

Influence of TiO2 Nanoparticle Filler on the Properties of PET and PLA Nanocomposites (이산화티탄 나노입자 필러가 PET와 PLA 나노복합체의 특성에 미치는 영향)

  • Farhoodi, Mehdi;Dadashi, Saeed;Mousavi, Seyed Mohammad Ali;Sotudeh-Gharebagh, Rahmat;Emam-Djomeh, Zahra;Oromiehie, Abdolrasul;Hemmati, Farkhondeh
    • Polymer(Korea)
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    • v.36 no.6
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    • pp.745-755
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    • 2012
  • Two types of polymers were tested in this study; poly(ethylene terephthalate) (PET) as a synthetic example and poly(lactic acid) (PLA) as a natural polymer. DSC analyses showed that the use of nanofiller increased the degree of crystallinity ($X_c$) of both PET and PLA polymers, but the effect was more noticeable on PET nanocomposites. The crystallization of PLA and PET nanocomposites occurred at higher temperatures in comparison to neat polymers. According to dynamic mechanical-thermal analysis (DMTA), the damping factor of PET/$TiO_2$ nanoparticles decreased compared to the neat matrix, but for PLA nanocomposites the opposite trend was observed. Results of the mechanical test showed that for both PET and PLA nanocomposites, the most successful toughening effect was observed at 3 wt% loading of $TiO_2$ nanoparticles. SEM micrographs revealed uniform distribution of $TiO_2$ nanoparticles at 1 and 3 wt% loading levels. The results of WAXD spectra explained that the polymorphs of PLA and PET was not affected by $TiO_2$ nanoparticles. UV-visible spectra showed that $TiO_2$ nanocomposite films had high ultraviolet shielding compared to neat polymer, but there was significant reduction in transparency.

The Production Mechanism of TNF-${\alpha}$ and IL-6 by Group IIA Phospholipase $A_2$ (IIA형 Phospholipase $A_2$에 의한 TNF-${\alpha}$와 IL-6 생성 기전)

  • Ahn, Jae-Hong;Park, Dae-Won;Kim, Jin-Hee;Bae, Jei-Jun;Bae, Yeun-Kyoung;Park, Yoon-Ki
    • Journal of Yeungnam Medical Science
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    • v.21 no.2
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    • pp.177-190
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    • 2004
  • Background: Secretory phospholipase $A_2$ ($sPLA_2$) are a group of extracellular enzymes that release fatty acids at the sn-2 position of phospholipids. Group IIA $sPLA_2$ ($sPLA_2$-IIA) has been detected in the inflammatory fluids, and its plasma level increases in the inflammatory disease. This study examined the effect of $sPLA_2$-IIA on mouse macropahges in order to investigate the potential mechanism of $sPLA_2$-induced inflammation. Materials and Methods: Wild type $PLA_2$ and mutant H48Q $PLA_2$ were purified from HEK293 cells transfected with the corresponding plasmids, and the $PLA_2$ activities were measured using 1-palmitoyl-2-[1-$^{14}C$]linoleoyl-3-phosphatidylethanolamine as substrates. The TNF-${\alpha}$ and IL-6 released in the supernatants were determined by ELISA. In addition, the TNF-${\alpha}$ and IL-6 mRNA were analyzed by RT-PCR. Results: $sPLA_2$-IIA stimulated the production of TNF-${\alpha}$ and IL-6 in a dose- and time-dependent manner. In addition, the effect of $sPLA_2$-IIA on cytokine production from the macrophage was found to be associated with the accumulation of their specific mRNA. The mRNA levels of TNF-${\alpha}$ and IL-6 peaked at 2 and 6 hours in a time-dependent manner, respectively. Conclusion: In conclusion, the production of proinflammatory cytokine might be mediated by the binding of $sPLA_2$-IIA to the receptors.

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Physical and electrical properties of PLA-carbon composites

  • Kang Z. Khor;Cheow K. Yeoh;Pei L. Teh;Thangarajan Mathanesh;Wee C. Wong
    • Advances in materials Research
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    • v.13 no.3
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    • pp.211-220
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    • 2024
  • Polylactic acid or polylactide (PLA) is a biodegradable thermoplastic that can be produced from renewable material to create various components for industrial purposes. In 3D printing technology, PLA is used due to its good mechanical, electrical, printing properties, environmentally friendly and non-toxic properties. However, the physical properties and excellent electrical insulation properties of PLA have limited its application. In this study, with the carbon black (CB) as filler added into PLA, the lattice spacing and morphology were investigated by using X-ray diffraction (XRD) and scanning electron microscope (SEM), respectively. The physical properties of PLA-carbon composite were evaluated by using tensile test, shore D hardness test and density and voids measurement. Impedance test was conducted to investigate the electrical properties of PLA-Carbon composites. The results demonstrate that the inclusion of carbon black as filler enhances the physical properties of the PLA-carbon composites, including tensile properties, hardness, and density. The addition of carbon black also leads to improved electrical conductivity of the composites. Better enhancement toward the electrical properties of PLA-carbon composites is observed with 1wt% of carbon black in N774 grade. The N550 grade with 2wt% of carbon black shows better improvement in the physical properties of PLA-carbon composites, achieving 10.686 MPa in tensile testing, 43.330 in shore D hardness test, and a density of 1.200 g/cm3 in density measurement. The findings suggest that PLA-carbon composites have the potential for enhanced performance in various industrial applications, particularly in sectors requiring improved physical and electrical properties.

Gene Expression Profiling by Ginsenoside Rb1 in Keratinocyte HaCaT Cells (피부각질세포 HaCaT에서 진세노사이드 Rb1에 의한 유전자 발현 양상)

  • Lee, Dong Woo;Kim, Jung Min;Bang, In Seok
    • Journal of Life Science
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    • v.29 no.5
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    • pp.514-523
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    • 2019
  • We investigated the gene expression patterns and the mechanisms of action of the apoptotic response by microarray analysis of human keratinocyte HaCaT cells treated with ginsenoside Rb1, a saponin of Panax ginseng C. A. Meyer. Genes related to apoptosis, the G2/M transition of the mitotic cell cycle, cell division, mitotic nuclear division, and intracellular protein transport were 2-fold up-regulated in HaCaT cells treated with the ginsenoside Rb1, whereas genes related to DNA repair, regeneration fission, and extracellular matrix organization were 2-fold down-regulated. Apoptosis signaling may be mediated by FAS and PLA2G4A, and pathway analysis indicated that STAT3 might be an upstream regulator of these genes. The activity of FAS and PLA2G4A was verified by qPCR, which showed that FAS was increased about 2-fold in HaCaT cells treated with $10{\mu}g/ml$ of ginsenoside Rb1 for 24 hr, PLA2G4A was increased about twice after 6 hours, and gene expression was increased more than 2-fold after 24 hr. Knockdown of STAT3 with siRNA decreased FAS expression and increased PLA2G4A expression but only FAS was passed from the upstream regulator STAT3. These results indicate that STAT3, which is an upstream regulator, induces apoptosis via FAS during treatment with ginsenoside Rb1.

Effect of PCB118 on expression of COX-2 and cPLA2 in rat testes

  • Han, Dae-Yong;Park, Kwang-Il;Park, Hyeon-Soo;Kang, Sang-Rim;Cho, Jae-Hyeon;Lee, Hu-Jang;Kim, Eun-Hee;Kim, Gon-Sup
    • Korean Journal of Veterinary Research
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    • v.49 no.4
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    • pp.345-349
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    • 2009
  • Polychlorinated biphenyls (PCBs) are synthetic organic compounds with two benzene rings and well known environmental pollutants. This study examined the effect of persistent exposure to 2,$3^{\prime}$,4,$4^{\prime}$,5-pentachlorobiphenyl (PCB118) on the proinflammatory and proapoptotic factors in male rats. Male Sprague Dawley rats were administered weekly intraperitoneal injections of either PCB118 (20 mg/kg) dissolved in corn oil or corn oil alone. One week after 2 and 5 administrations, the rats were sacrificed by a pentobarbital injection. The effect of PCB118 on the expression of cyclooxygenase 2 (COX-2), cytosolic phospholipase A2 (cPLA2), peroxisome proliferator-activated receptor gamma, Bcl and Bcl-2-associated X protein (BAX) was investigated. The level of COX-2 and cPLA2 expression was higher in the PCB118-treated rats than the control. These results suggest that PCB118 has a proinflammatory effect in rats.

Modulation of Uterine Phospholipase $A_2$ Activity by Estradiol During the Delayed Implantation Process in Rats (흰쥐의 착상기간중 Estradiol이 자궁의 Phospholipase $A_2$ 활성도에 미치는 영향)

  • Yoon, Mi-Chung;Kim, Chang-Mee;Choe, Rim-Soon;Ryu, Kyung-Za
    • The Korean Journal of Pharmacology
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    • v.27 no.2
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    • pp.191-196
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    • 1991
  • The present study was performed to determine whether estradiol, via cAMP mediation, induces prostaglandin synthesis by modulating phospholipase $A_2$ activity which hydrolyzes phospholipids into arachidonic acids, a precursor for prostaglandin synthesis, during the implantation process in rats. Uterine phospholipase $A_2$ activity was elevated on day 5 of pregnancy when implantation normally occurs in rats. Moreover, phospholipase $A_2$ activity was higher in the implant sites than in the non-implant sites of uterus on day 6. In delayed implantation model, phospholipase $A_2$ activity was increased at 12 hrs after estradiol administration and at 8 hrs after dbcAMP administration. In addition, higher activity of phospholipase $A_2$ was induced by the treatment of estradiol plus theophylline, compared with estradiol-only treated group. The simultaneous treatment of indomethacin with estradiol or dbcAMP did not alter phospholipase $A_2$ activity compared with estradiol or dbcAMP-only treated group although significant suppression was observed in uterine PGE and $PGF_{2{\alpha}}$ concentrations. These results suggest that estradiol or cAMP stimulates uterine phospholipase $A_2$ activity, thereby increasing prostaglandin synthesis during the implantation process in rats.

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Biomechanical Analysis of Biodegradable Cervical Plates Developed for Anterior Cervical Discectomy and Fusion

  • Cho, Pyung Goo;Ji, Gyu Yeul;Park, Sang Hyuk;Shin, Dong Ah
    • Asian Spine Journal
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    • v.12 no.6
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    • pp.1092-1099
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    • 2018
  • Study Design: In-vitro biomechanical investigation. Purpose: To evaluate the biomechanical effects of the degeneration of the biodegradable cervical plates developed for anterior cervical discectomy and fusion (ACDF) on fusion and adjacent levels. Overview of Literature: Biodegradable implants have been recently introduced for cervical spine surgery. However, their effectiveness and safety remains unclear. Methods: A linear three-dimensional finite element (FE) model of the lower cervical spine, comprising the C4-C6 vertebrae was developed using computed tomography images of a 46-year-old woman. The model was validated by comparison with previous reports. Four models of ACDF were analyzed and compared: (1) a titanium plate and bone block (Tita), (2) strong biodegradable plate and bone block (PLA-4G) that represents the early state of the biodegradable plate with full strength, (3) weak biodegradable plate and bone block (PLA-1G) that represents the late state of the biodegradable plate with decreased strength, and (4) stand-alone bone block (Bloc). FE analysis was performed to investigate the relative motion and intervertebral disc stress at the surgical (C5-C6 segment) and adjacent (C4-C5 segment) levels. Results: The Tita and PLA-4G models were superior to the other models in terms of higher segment stiffness, smaller relative motion, and lower bone stress at the surgical level. However, the maximal von Mises stress at the intervertebral disc at the adjacent level was significantly higher in the Tita and PLA-4G models than in the other models. The relative motion at the adjacent level was significantly lower in the PLA-1G and Bloc models than in the other models. Conclusions: The use of biodegradable plates will enhance spinal fusion in the initial stronger period and prevent adjacent segment degeneration in the later, weaker period.