• Title/Summary/Keyword: LPS transfer

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Dynamic lipopolysaccharide transfer cascade to TLR4/MD2 complex via LBP and CD14

  • Kim, Soo Jin;Kim, Ho Min
    • BMB Reports
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    • v.50 no.2
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    • pp.55-57
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    • 2017
  • Toll-like receptor 4 (TLR4) together with MD2, one of the key pattern recognition receptors for a pathogen-associated molecular pattern, activates innate immunity by recognizing lipopolysaccharide (LPS) of Gram-negative bacteria. Although LBP and CD14 catalyze LPS transfer to the TLR4/MD2 complex, the detail mechanisms underlying this dynamic LPS transfer remain elusive. Using negative-stain electron microscopy, we visualized the dynamic intermediate complexes during LPS transfer-LBP/LPS micelles and ternary CD14/LBP/LPS micelle complexes. We also reconstituted the entire cascade of LPS transfer to TLR4/MD2 in a total internal reflection fluorescence (TIRF) microscope for a single molecule fluorescence analysis. These analyses reveal longitudinal LBP binding to the surface of LPS micelles and multi-round binding/unbinding of CD14 to single LBP/LPS micelles via key charged residues on LBP and CD14. Finally, we reveal that a single LPS molecule bound to CD14 is transferred to TLR4/MD2 in a TLR4-dependent manner. These discoveries, which clarify the molecular mechanism of dynamic LPS transfer to TLR4/MD2 via LBP and CD14, provide novel insights into the initiation of innate immune responses.

Effect of Lipopolysaccharides Treatment on Uterine Involution in Dairy Cows with Retained Placenta (후산정체 젖소에 있어서 Lipopolysaccharide의 처리가 자궁 회복에 미치는 영향)

  • Baek, K.S.;Park, S.J.;Park, S.B.;Lee, W.S.;Kim, H.S.;Jeong, G.Y.;Ki, K.S.;Suh, G.H.;Jeon, B.S.;Ahn, B.S.;Kim, T.I.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.233-239
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    • 2006
  • This study was carried out to investigate the effect of intra-uterine infusion of lipopolysaccharides (LPS) on uterine involution in dairy cows with retained placenta. The LPS isolated from Bacteroides helcogenes and Fusobaoterium varium was infused at the rate of 100 ug (n=11) and 200 ug (n=11) with 30 ml of phosphate buffer saline in each cow at 20 days post-partum. The cows in conrol group (n=11) with retained placenta were infused with 30 ml phosphate buffer saline in each cow at 20 days postpartum. Cross-sectional ultrasonography was performed at days 40 postpartum to examine uterine involution and exudate in the all dairy cows in study. The cows in control group have shown 45.5, 27.3 and 27.3% gravid horn with less than 30 mm (normal), $31{\sim}50mm$ (medium) and more than 51 mm (large) in diameter. Respective average values of gravid horn diameter in cows treated with LPS 200 ug were noticed 72.7, 18.2 and 9.1%. However, the gravid horn diameter in all the cows treated with 100 ug of LPS was less than 30 mm. The 18.2, 63.6 and 54.6% cows in the control, LPS 100 ug and LPS 200 ug, respectively, have shown no exudate in uterine cavity. However, the respective rates of small amount of exudate retention in uterine cavity were 45.5, 0.0, and 9.1%. Approximately 36.4% cows in all 3 treated groups have shown minute quantity of exudate. In conclusion, the LPS intra-uterine infusion promoted postpartum uterine involution in dairy cows with retained placenta.

Anti-Endotoxin 9-Meric Peptide with Therapeutic Potential for the Treatment of Endotoxemia

  • Krishnan, Manigandan;Choi, Joonhyeok;Choi, Sungjae;Kim, Yangmee
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.25-32
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    • 2021
  • Inflammatory reactions activated by lipopolysaccharide (LPS) of gram-negative bacteria can lead to severe septic shock. With the recent emergence of multidrug-resistant gram-negative bacteria and a lack of efficient ways to treat resulting infections, there is a need to develop novel anti-endotoxin agents. Antimicrobial peptides have been noticed as potential therapeutic molecules for bacterial infection and as candidates for new antibiotic drugs. We previously designed the 9-meric antimicrobial peptide Pro9-3 and it showed high antimicrobial activity against gram-negative bacteria. Here, to further examine its potency as an anti-endotoxin agent, we examined the anti-endotoxin activities of Pro9-3 and elucidated its mechanism of action. We performed a dye-leakage experiment and BODIPY-TR cadaverine and limulus amebocyte lysate assays for Pro9-3 as well as its lysine-substituted analogue and their enantiomers. The results confirmed that Pro9-3 targets the bacterial membrane and the arginine residues play key roles in its antimicrobial activity. Pro9-3 showed excellent LPS-neutralizing activity and LPS-binding properties, which were superior to those of other peptides. Saturation transfer difference-nuclear magnetic resonance experiments to explore the interaction between LPS and Pro9-3 revealed that Trp3 and Tlr7 in Pro9-3 are critical for attracting Pro9-3 to the LPS in the gram-negative bacterial membrane. Moreover, the anti-septic effect of Pro9-3 in vivo was investigated using an LPS-induced endotoxemia mouse model, demonstrating its dual activities: antibacterial activity against gram-negative bacteria and immunosuppressive effect preventing LPS-induced endotoxemia. Collectively, these results confirmed the therapeutic potential of Pro9-3 against infection of gram-negative bacteria.

Effect of Treatment of Lipopolysaccharides on Reproductive Performance in Lactating Dairy Cows (젖소에 있어서 Lipopolysaccharide의 처리가 번식 성적에 미치는 영향)

  • Baek K.S.;Park, S.B.;Park, S.J.;Kim, H.S.;Lee, W.S.;Jeong, G.Y.;Lee, H.J.;Hur, T.Y.;Jeon B.S.;Ahn B.S.;Khan M. Aiumal
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.241-245
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    • 2006
  • This study was carried out to investigate the effect of intra-uterine infusion of lipopolysaccharide (LPS) on reproductive performance in lactating dairy cows. Conception rate at first service was higher (56.0%) in dairy cows treated with LPS compared to non-treated cows (20.0 %). Cumulative conception rate by overall services (more than 2 services) after LPS treatment was also higher (64.0%) in dairy cows treated with LPS compared to non-treated cows (40.0%). Services per conception was significantly higher in control ($2.0{\pm}0.1$) compared with LPS ($1.2{\pm}0.4$) treated group (p<0.05).

Lipooligosaccharide biosynthesis genes of nontypable Haemophilus influenzae 2019

  • Lee, Na-Gyong;Melvin G. Sunshine;Jeffery J. Engstrom;Bradford W. Gibson;Michael A. Apicella
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.11a
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    • pp.65-73
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    • 1996
  • LPS/LOS, the compound found only in gram-negative bacterial outer membrane, plays important roles in bacterial maintenance as well as its pathogenesis. We isolated and characterized several genes required for NTHi 2019 LOS biosynthesis, which encode enzymes required for sugar substrate synthesis or the transfer of substrates to receptor molecules. The htrB gene, however, appears to have more complex role. It has acryltransferase activity as well as various other activity, which may control regulation of LOS biosynthesis as well as its pathogenicity. Evidences supporting the latter come from the observations that the lipid A of the B29 induced significantly less TNF ${\alpha}$ from macrophages than that of the wild type LOS (unpublished data). H. influenzae A2-htrB mutant strain was also significantly less invasive than the wild type strain. The structural similarities of the enterobacterial LPS and the Haemophilus LOS enabled us to isolate the NTHi 2019 genes involved in LOS biosynthesis genes by using the S. typhimurium LPS deep core mutants. While a similar approach has been used for E. coli, this technique for selection of an LPS phenotype has not been applied to nonenterobacterial species. The difficulties inherent in the molecular manipulation of organism such as Neisseria and Haemophilus species make this approach particularly attractive in the identification and cloning LOS genes. Studies on genetic features of LPS/LOS biosynthesis would be useful for understanding bacterial pathogenesis as well as for developing vaccines for these gram-negative pathogenic bacteria.

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Effect of luteal phase support with vaginal progesterone on pregnancy outcomes in natural frozen embryo transfer cycles: A meta-analysis

  • Seol, Aeran;Shim, Yoo Jin;Kim, Sung Woo;Kim, Seul Ki;Lee, Jung Ryeol;Jee, Byung Chul;Suh, Chang Suk;Kim, Seok Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.2
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    • pp.147-152
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    • 2020
  • Objective: The purpose of this study was to determine the effect of vaginal progesterone for luteal phase support (LPS) on the clinical pregnancy rate (CPR) in natural frozen embryo transfer (FET) cycles via a meta-analysis. Methods: We performed a meta-analysis of randomized controlled trials (RCTs) and retrospective studies that met our selection criteria. Four online databases (PubMed, Embase, Medline, and the Cochrane Library) were searched between January 2017 and May 2017. Studies were selected according to predefined inclusion criteria and meta-analyzed using R software version 2.14.2. The main outcome measure was CPR. Results: A total of 18 studies were reviewed and assessed for eligibility. One RCT (n = 435) and three retrospective studies (n = 3,033) met the selection criteria. In a meta-analysis of the selected studies, we found no significant difference in the CPR (odds ratio [OR], 0.96; 95% confidence interval [CI], 0.60-1.55) between the vaginal progesterone and control groups. An analysis of the two retrospective cohort studies that reported the live birth rate (LBR) following FET showed a significantly higher LBR in the vaginal progesterone group (OR, 1.72; 95% CI, 1.21-2.46). A subgroup meta-analysis of FET conducted 5 days after injection of human chorionic gonadotropin showed no significant differences between the two groups with regard to the CPR (OR, 1.18; 95% CI, 0.90-1.55) or miscarriage rate (OR, 0.73; 95% CI, 0.36-1.47). Conclusion: The results of this meta-analysis of the currently available literature suggest that LPS with vaginal progesterone in natural FET cycles does not improve the CPR.

Understanding of the Duplex Thrust System - Application to the Yeongwol Thrust System, Taebaeksan Zone, Okcheon Belt (듀플렉스트러스트시스템의이해 - 옥천대태백산지역영월트러스트시스템에의 적용)

  • Jang, Yirang
    • Economic and Environmental Geology
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    • v.52 no.5
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    • pp.395-407
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    • 2019
  • The duplex system has been considered as an important slip-transfer mechanism to evaluate the evolution of orogenic belts. Duplexes are generally found in the hinterland portion of fold-thrust belts and accommodate large amounts of total shortening. Thus, understanding its geometric and kinematic evolution can give information to evaluate the evolution of the entire orogenic belt. Duplexes are recognized as closed-loop thrust traces on map view, indicating higher connectivity than imbricate fans. As originally defined, a duplex is an array of thrust horses which are surrounded by thrust faults including the floor and roof thrusts, and imbricate faults between them. Duplexes can accommodate regional layer-parallel shortening and transfer slip from a floor thrust to a roof thrust. However, an imbricate fault is not the only mean for layer-parallel shortening (LPS) and displacement transfer within duplexes. LPS cleavages and detachment folds can also play the same role. From this aspect, a duplex can be divided into three types; 1) fault duplex, 2) cleavage duplex and 3) fold duplex. Fault duplex can further be subdivided into the Boyer-type duplex, which was firstly designed duplex system in the 1980s that widely applied most of the major fold-thrust belts in the world, and connecting splay duplex, which has different time order in the emplacement of horses from those of the Boyer-type. On the contrary, the cleavage and fold duplexes are newly defined types based on some selected examples. In the Korean Peninsula, the Yeongwol area, the western part of the Taebaeksan Zone of the Okcheon Belt, gives an excellent natural laboratory to study the structural geometry and kinematics of the closed-loops by thrust fault traces in terms of a duplex system. In the previous study, the Yeongwol thrust system was interpreted by alternative duplex models; a Boyer-type hinterland-dipping duplex vs. a combination of major imbricate thrusts and their connecting splays. Although the high angled beds and thrusts as well as different stratigraphic packages within the horses of the Yeongwol duplex system may prefer the later complicate model, currently, we cannot choose one simple answer between the models because of the lack of direct field evidence and time information. Therefore, further researches on the structural field investigations and geochronological analyses in the Yeongwol and adjacent areas should be carried out to test the possibility of applying the fold and cleavage duplex models to the Yeongwol thrust system, and it will eventually provide clues to solve the enigma of formation and its evolution of the Okcheon Belt.

Passive Immunity by Splenocyte Transfer against Amebic Meningoeneephalitis in Mice (세포에 의한 아메바성 수막뇌염에 대한 피동면역의 전달)

  • 임경일;유재숙
    • Parasites, Hosts and Diseases
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    • v.26 no.3
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    • pp.169-174
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    • 1988
  • The role of passive cell-mediated transfer of immunity against primary amoebic meningoen- cephalitis(PAME) in mice was studied. Waegleria fowleri, ITMAP 359, were cultured in CGVS medium. The ICR mice used were six week-old males of average weight of 15 g. Immunization was done by three intraperitoneal injections of $1{\times}10^6$ N. fowleri trophozoites at the interval of one week. Splenocytes were obtained from normal and immune mice spleens, and Ix107 cells were administered intraperitoneally into mice 3 days before challenge infection. Mice were infected intranasally with $7{\times}10^4$ N. fowleri trophozoites in a $3{\;}{\mu}l$ suspension under secobarbiturate anesthesia. Transplants of normal or immune splenocytes seem to alter the pattern of the PAME level- opment. The splenocytcs transferred from immune mice reduced the mortality rate in the JV. fowleri infected mice, as compared with the mice transferred with the same number of normal splenocytes or without splenocyte, The blastogenic response of the splenocytes to both lipopoly- saccharide and concanavalin A was elevated on duty 7 after infection the mice transinoculated with immune splenocytes. The serum antibody titers in the mice transferred with immune spleno- cytes were increased gradually from day 7 up to day 20 after infections by mean of ELISA. It is suggested that the transfer of splenocytes from immuniged mice conferred immunity against N. fowleri infection.

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Binding Model of Fisetin and Human c-Jun NH2-Terminal Kinase 1 and Its Anti-inflammatory Activity

  • Jnawali, Hum Nath;Lee, Eunjung;Jeong, Ki-Woong;Heo, Yong-Seok;Kim, Yangmee
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2629-2634
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    • 2013
  • Fisetin is a naturally occurring flavonoid with some anti-cancer and anti-inflammation capabilities. In this study, we perform docking studies between human c-Jun N-terminal kinase 1 (JNK 1) and fisetin and proposed a binding model of fisetin and JNK 1, in which the hydroxyl groups of the B ring and oxygen at the 4-position of the C ring play key roles in binding interactions with JNK. Fluorescence quenching and saturation-transfer difference (STD) NMR experiments showed that fisetin exhibits good binding affinity to JNK, $1.32{\times}10^8M^{-1}$. The anti-inflammatory activity of fisetin was also investigated. Fisetin significantly suppressed tumor necrosis factor, the NO production, and macrophage inflammatory cytokine release in LPS-stimulated RAW264.7 mouse macrophages. We found that the anti-inflammatory cascade of fisetin was mediated through the JNK, and cyclooxygenase (COX)-2 pathways. Our findings suggest the potential of fisetin as an anti-inflammatory agent.