• Title/Summary/Keyword: lipopolysaccharides (LPS)

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Depression of L-type $Ca^{2+}$ and Transient Outward $K^+$ Currents in Endotoxin-treated Rat Cardiac

  • Park, Kyu-Sang;Lee, Boo-Soo;Kong, In-Deok;Lee, Joong-Woo
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.6
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    • pp.623-630
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    • 1999
  • Decreased cardiac contractility occurs in endotoxicosis, but little is known about the ionic mechanism responsible for myocardial dysfunction. In this study, we examined the changes in $Ca{2+}$ and $K^+$ currents in cardiac myocytes from endotoxin-treated rat. Ventricular myocytes were isolated from normal and endotoxemic rats (ex vivo), that were treated for 10 hours with Salmonella enteritidis lipopolysaccharides (LPS; 1.5 mg/kg) intravenously. Normal cardiac myocytes were also incubated for 6 hours with 200 ng/ml LPS (in vitro). L-type $Ca{2+}$ current $(I_{Ca,L})$ and transient outward $K^+$ current $(I_{to})$ were measured using whole cell patch clamp techniques. Peak $I_{Ca,L}$ was reduced in endotoxemic myocytes (ex vivo; 6.00.4 pA/pF, P<0.01) compared to normal myocytes (control; 10.90.6 pA/pF). Exposure to endotoxin in vitro also attenuated $I_{Ca,L}$ (8.40.4 pA/pF, P<0.01). The amplitude of $(I_{to})$ on depolarization to 60 mV was reduced in endotoxin treated myocytes (16.51.5 pA/pF, P<0.01, ex vivo; 20.00.9 pA/pF, P<0.01 , in vitro) compared to normal myocytes (control; 24.71.0 pA/pF). There was no voltage shift in steady-state inactivation of $I_{Ca,L}$ and $(I_{to})$ between groups. These results suggest that endotoxin reduces $Ca{2+}$ and $K^+$ currents of rat cardiac myocytes, which may lead to cardiac dysfunction.

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Mild encephalopathy with a reversible splenial lesion in a girl with acute pyelonephritis

  • Yeom, Jung Sook;Koo, Chung Mo;Park, Ji Sook;Seo, Ji-Hyun;Park, Eun Sil;Lim, Jae-Young;Woo, Hyang-Ok;Youn, Hee-Shang
    • Clinical and Experimental Pediatrics
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    • v.61 no.2
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    • pp.64-67
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    • 2018
  • We report the case of a 12-year-old girl who had mild encephalopathy with a reversible splenial lesion (MERS) associated with acutepyelonephritis caused by Escherichia coli. The patient was admitted with a high fever, and she was diagnosed with acute pyelonephritis based on pyuria and the results of urine culture, which detected cefotaxime-sensitive E. coli. Although intravenous cefotaxime and tobramycin were administered, her fever persisted and her C-reactive protein level increased to 307 mg/L. On day 3 of admission, she demonstrated abnormal neuropsychiatric symptoms, such as delirium, ataxia, and word salad. Magnetic resonance imaging (MRI) of the brain performed on day 4 showed marked hyperintensities in the bilateral corpus callosum and deep white matter on diffusion-weighted images, with corresponding diffusion restriction on apparent diffusion coefficient mapping. No abnormalities or pathogens were detected in the cerebrospinal fluid; however, lipopolysaccharides (LPS, endotoxin) were detected in plasma (41.6 pg/mL), associated with acute neurological deterioration. Her clinical condition gradually improved, and no neurological abnormalities were observed on day 6. Follow-up brain MRI performed 2 weeks later showed near-disappearance of the previously noted hyperintense lesions. In this patient, we first proved endotoxemia in a setting of MERS. The release of LPS following antibiotic administration might be related to the development of MERS in this patient. The possibility of MERS should be considered in patients who present with acute pyelonephritis and demonstrate delirious behavior.

Characterization of Hemolytic Aeromonas sp. MH-8 Responding to the Exposure of Green Tea Catechin, EGCG (녹차 카테킨 EGCG의 노출에 따른 식중독 세균인 용혈성 Aeromonas sp. MH-8의 특성조사)

  • Kim, Dong-Min;Oh, Kye-Heon
    • KSBB Journal
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    • v.31 no.4
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    • pp.228-236
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    • 2016
  • The aim of this study was to characterize the hemolytic Aeromonas sp. MH-8 exposed to green tea catechin, epigallocatechin gallate (EGCG). Initially, the hemolytic Aeromonas sp. MH-8 was enriched and isolated from stale fish. Bactericidal effects of MH-8 exposed to EGCG ranging from 1 mg/mL to 4 mg/mL were monitored, and complete bactericidal effects were achieved within 3 h at 3 mg/mL and higher concentrations. SDS-PAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain MH-8 treated to different concentrations and exposing periods of EGCG in exponentially growing cultures. The stress shock proteins (70-kDa DnaK and 60-kDa GroEL), which might contribute to enhancing the cellular resistance to the cytotoxic effect of EGCG, were induced at different concentrations of EGCG exposed to cell culture of MH-8. Scanning electron microscopic analysis demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with EGCG. 2-DE of soluble protein fractions from MH-8 cultures showed 18 protein spots changed by EGCG exposure. These proteins involved in chaperons (e.g., DnaK, GroEL and trigger factor), enterotoxins (e.g., aerolysin and phospholipase C precursor), LPS synthesis (e.g., LPS biosynthesis protein and outer membrane protein A precursor), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. In consequence, EGCG was found to have substantial antibacterial effects against food-poisoning causing bacterium, hemolytic Aeromonas sp. MH-8. Also the results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on Aeromonas sp. MH-8.

Biological effects of a root conditioning agent for dentin surface modification in vitro

  • Lee, Jue-Yeon;Seol, Yang-Jo;Park, Jang-Ryul;Park, Yoon-Jeong;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.40 no.6
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    • pp.257-264
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    • 2010
  • Purpose: Connective tissue reattachment to periodontally damaged root surfaces is one of the most important goals of periodontal therapy. The aim of this study was to develop a root conditioning agent that can demineralize and detoxify the infected root surface. Methods: Dentin slices obtained from human teeth were treated with a novel root planing agent for 2 minutes and then washed with phosphate-buffered saline. Smear layer removal and type I collagen exposure were observed by scanning electron microscopy (SEM) and type I collagen immunostaining, respectively. Cell attachment and lipopolysaccharides (LPS) removal demonstrated the efficiency of the root conditioning agent. Results: SEM revealed that the smear layer was entirely removed and the dentinal tubules were opened by the experimental gel. Type I collagen was exposed on the surfaces of the dentin slices treated by the experimental gel, which were compared with dentin treated with other root planing agents. Dentin slices treated with the experimental gel showed the highest number of attached fibroblasts and flattened cell morphology. The agar diffusion assay demonstrated that the experimental gel also has effective antimicrobial activity. Escherichia coli LPS were effectively removed from well plates by the experimental gel. Conclusions: These results demonstrated that this experimental gel is a useful tool for root conditioning of infected root surfaces and can also be applied for detoxification of ailing implant surface threads.

Effects of Butanol extract from Rhois Vernicifluae Cortex (RVC) in lipopolysaccharides-induced macrophage RAW 264.7 cells (칠피(漆皮) 부탄올 분획물이 LPS로 유도된 RAW 264.7 대식세포에 미치는 영향)

  • Song, Saeng-Yeop;Sim, Sung-Yong;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.20 no.1 s.32
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    • pp.1-15
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    • 2007
  • Objectives : RVC has long been used for a useful natural agent ameliorating inflammation related symptoms in the folk medicine recipe. This study was performed to investigate effects of RVC on the inflammation and oxidation in RAW 264.7 cells. Methods : The RVC was extracted with 80% ethanol and sequentially partitioned with solvents in order to increase polarity. With the various fractions, we determined the activities on the inflammation and oxidation in RAW 264.7 cells. Results : 1. Among the various solvent extracts of RVC, the butanol fraction showed the most powerful inhibitory ability against nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells without affecting cell viability. 2. Butanol fraction showed a oxidation inhibition effect by decreasing the DPPH and OH radicals. 3. Butanol fraction exhibited the inhibitory avilities against iNOS and COX-2. 4. Reverse transcriptase polymerase chain reaction (RT-PCR) and Westem blotting analysis revealed that the BuOH fraction provided a primary inhibitor of the iNOS protein and mRNA expression in LPS-induced RAW 264.7 cells. Among the up-regulater molecules of iNOS and COX-2, the BuOh fraction of RVC was shown the inhibitory activity of phoshporylation of c-Jun N-terminal kinase (JNK) 1/2 and threonine protein kinase (AKT), the one of the MAPKs pathway. Conclusion : Thus, the present study suggests that the response of a component of the BuOH fraction to NO generation via iNOS expression provide a important clue to elucidate anti-inflammatory and anti-oxidation mechanism of RVC.

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Antibacterial Activity against Salmonella enteritidis JK-15 and LPS Changes Caused by Rose Flower Extracts (장미꽃 추출물에 의한 식중독 세균 Salmonella enteritidis JK-15에 대한 살균활성 및 그에 따른 LPS 변화)

  • Song, You-Jin;Cho, Yun-Seok;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.318-323
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    • 2009
  • The aim of this work was to investigate the antibacterial effect of the food-poisoning bacterium, Salmonella enteritidis JK-15 exposed to rose extracts. Initially, the isolate S. enteritidis JK-15 was enriched and isolated from stale food. BIOLOG and 16S rRNA analyses revealed that strain S. enteritidis JK-15 was 98% similar to the S. enteritidis species cluster; therefore we have designated this strain as S. enteritidis JK-15. Bactericidal effects of S. enteritidis JK-15 exposed to rose extracts ranging from 5 mg/ml to 100 mg/ml were monitored, and complete bactericidal effects were achieved within 6 h at 100 mg/ml and 12 h at 50 mg/ml, respectively. SDSPAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain S. enteritidis JK-15 treated to different concentrations and exposing periods of rose extracts in exponentially growing cultures. Scanning electron microscopic analysis, demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with rose extracts.

Analysis of Anti-inflammatory Efficacy of Chrysanthemum coronarium PDRN (Polydeoxyribonucleotide) (쑥갓(Chrysanthemum coronarium L.) PDRN(Polydeoxyribonucleotide)의 항염증 효능 분석)

  • Song, Mi-Hee;Choi, Moon-Hyeok;Jeong, Jin-Hyoung;Lee, Sang-Sik;Jeong, Woo-Young
    • The Journal of Korea Institute of Information, Electronics, and Communication Technology
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    • v.15 no.5
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    • pp.396-404
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    • 2022
  • Chrysanthemum coronarium L. contains various antioxidants such as beta-carotene, vitamins A and C, and polyphenols, and is known to have anti-inflammatory effects. Under the assumption that the PDRN contained in the extract can mediate the anti-inflammatory response, the mouse macrophages, RAW264.7 cells, were stimulated with LPS to induce the conversion to inflammatory cells, and then the addition of PDRN extracted from the extract was effective in inhibiting inflammation. It was analyzed whether there was The gene expression of IL-1β and TNF-α was used as an anti-inflammatory index, and the relative expression levels of each gene were confirmed by RT-PCR. As a result, RT-PCR confirmed the effect of PDRN-induced inhibition of inflammation in both IL-1β and TNF-α genes. Therefore, based on this study, it is considered to be a precedent data that can be developed as a treatment for inflammatory diseases, and it will be helpful in research and development of a treatment that can improve the anti-inflammatory mechanism.

Anti-inflammation Effect of Cyrtomium fortunei J.Sm. Extracts in Lipopolysaccharides-induced Microglia BV2 Cell (LPS로 자극한 microglia BV2 cell에서 Cyrtomium fortunei J.Sm. 추출물의 항염증 효과)

  • Jiwon Choi;Shintae Kim;Sang Yoon Choi;Inwook Choi;Jinyoung Hur
    • Journal of the Korean Society of Food Culture
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    • v.38 no.3
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    • pp.176-183
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    • 2023
  • In this study, we investigated the effect of the extracts of Cyrtomium fortunei J.Sm. (CFJ) on lipopolysaccharide (LPS) induced inflammation in mouse BV-2 microglial cells. Nitric oxide (NO) production and cell viability were measured using the Griess reagent and the (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) (MTT) assay. Inflammatory cytokines were detected by quantitative polymerase chain reaction (qPCR) in BV-2 microglial cells with and without CFJ extracts. Subsequently, mitogen-activated protein kinases (MAPKs) and antioxidant markers were assessed by western blot analysis. It was found that the CFJ extract significantly decreased the production of pro-inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor [TNF]-α, and IL-1β) and NO in BV-2 microglial cells that were stimulated with LPS. In addition, the expression levels of the phosphorylation of the MAPK family (p38, c-Jun N-terminal kinases [JNK], and extracellular-signal regulated kinase [ERK]) were reduced by CFJ. Also, treatment with CFJ significantly increased the activities of superoxide dismutase type 1(SOD1) and Catalase in BV-2 microglial cells. Our results indicate that CFJ has a potent suppressive effect on the pro-inflammatory responses of activated BV-2 microglia. Therefore, CFJ has the potential to be an effective treatment for neurodegenerative diseases, as it can inhibit the production of inflammatory mediators in activated BV-2 microglial cells.

Ginsenoside Rd inhibits the expressions of iNOS and COX-2 by suppressing NF-κB in LPS-stimulated RAW264.7 cells and mouse liver

  • Kim, Dae Hyun;Chung, Jae Heun;Yoon, Ji Sung;Ha, Young Mi;Bae, Sungjin;Lee, Eun Kyeong;Jung, Kyung Jin;Kim, Min Sun;Kim, You Jung;Kim, Mi Kyung;Chung, Hae Young
    • Journal of Ginseng Research
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    • v.37 no.1
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    • pp.54-63
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    • 2013
  • Ginsenoside Rd is a primary constituent of the ginseng rhizome and has been shown to participate in the regulation of diabetes and in tumor formation. Reports also show that ginsenoside Rd exerts anti-oxidative effects by activating anti-oxidant enzymes. Treatment with ginsenoside Rd decreased nitric oxide and prostaglandin $E_2$ ($PGE_2$) in lipopolysaccharides (LPS)-challenged RAW264.7 cells and in ICR mouse livers (5 mg/kg LPS; LPS + ginsenoside Rd [2, 10, and 50 mg/kg]). Furthermore, these decreases were associated with the down-regulations of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 and of nuclear factor (NF)-${\kappa}B$ activity in vitro and in vivo. Our results indicate that ginsenoside Rd treatment decreases; 1) nitric oxide production (40% inhibition); 2) $PGE_2$ synthesis (69% to 93% inhibition); 3) NF-${\kappa}B$ activity; and 4) the NF-${\kappa}B$-regulated expressions of iNOS and COX-2. Taken together, our results suggest that the anti-inflammatory effects of ginsenoside Rd are due to the down-regulation of NF-${\kappa}B$ and the consequent expressional suppressions of iNOS and COX-2.

Anti-inflammatory Effect of Gagam-GongJin-dan in mouse peritoneal macrophages (마우스 복강대식세포에서 가감공진단(加減拱辰丹)의 항염증 효과)

  • Kim, Hong-Jun;Kim, Young-Sik;Mok, Ji-Ye;Jeong, Seung-Il;Hwang, Sung-Yeoun;Cho, Jung-Keun;Jang, Seon-Il
    • Herbal Formula Science
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    • v.19 no.1
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    • pp.207-217
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    • 2011
  • Objectives : In a previous study, we have shown that Gagam-Gongjin-Dan(GGD) has an inhibitory effect on the ovalbumin-induced immune responses and a hepatoprotective effect on actaminophen-induced liver injury in Balb/c Mice. However, the possible anti-inflammatory effect of GGD extract for inflammatory mediators was not reported. Therefore, the purpose of this study was to investigate an inhibitory effects of GGD extract against lipopolysaccharides(LPS) induced inflammatory mediators in mouse peritoneal macrophages. Methods : GGD extract was prepared by extracting with methanol for 7 days. The extract was freeze-dried following filtration through vacuum distillation system. Accumulated nitrite, an oxidative product of nitric oxide(NO), was measured in the culture medium by the Griess reaction. The levels of prostaglandin $E_2(PGE_2)$, interleukin-$1{\beta}$(IL-$1{\beta}$), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) were measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2(COX-2) were measured by Western blot analysis. Results : GGD extract (50-$400\;{\mu}g$/ml) per se had no cytotoxic effect in LPS-stimulated peritoneal macrophages. GGD extract dose-dependently reduced NO, $PGE_2$, IL-$1{\beta}$ and TNF-${\alpha}$ production and COX-2 activity caused by stimulation of LPS. The levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with GGD extract in a dose dependent manner. Conclusions : These results suggest that GGD extract has an anti-inflammatory effect against LPS-induced inflammatory mediators in peritoneal macrophages, these properties may contribute to inflammation disease care.