• Title/Summary/Keyword: Mice model

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Real-time Imaging of Inositol 1,4,5-trisphosphate Movement in Mouse Salivary Gland Cells

  • Hong, Jeong-Hee;Lee, Syng-Ill;Shin, Dong-Min
    • International Journal of Oral Biology
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    • v.33 no.4
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    • pp.125-129
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    • 2008
  • Inositol 1,4,5-trisphosphate ($IP_3$) plays an important role in the release of $Ca^{2+}$ from intracellular stores into the cytoplasm in a variety of cell types. $IP_3$ translocation dynamics have been studied in response to many types of cell signals. However, the dynamics of cytosolic $IP_3$ in salivary acinar cells are unclear. A green fluorescent protein (GFP)-tagged pleckstrin homology domain (PHD) was constructed and introduced into a phospholipase C ${\delta}1$ (PLC ${\delta}1$) transgenic mouse, and then the salivary acinar cells were isolated. GFP-PHD was heterogeneously localized at the plasma membrane and intracellular organelles in submandibular gland and parotid gland cells. Application of trypsin, a G protein-coupled receptor activator, to the two types of cells caused an increase in GFP fluorescence in the cell cytoplasm. The observed time course of trypsin-evoked $IP_3$ movement in acinar cells was independent of cell polarity, and the fluorescent label showed an immediate increase throughout the cells. These results suggest that GFP-PHD in many tissues of transgenic mice, including non-cultured primary cells, can be used as a model for examination of $IP_3$ intracellular dynamics.

Nonribosomal Peptide Synthase is Responsible for the Biosynthesis of Siderophore in Vibrio vulnificus MO6-24/O

  • Kim, In-Hwang;Shim, Jung-Im;Lee, Ko-Eun;Hwang, Won;Kim, Ik-Jung;Choi, Sang-Ho;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.35-42
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    • 2008
  • Vibrio vulnificus produces siderophores, low-molecular-weight iron-chelating compounds, to obtain iron under conditions of iron deprivation. To identify genes associated with the biosynthesis of siderophore in V. vulnificus MO6-24/O, we screened clones with mini-Tn5 random insertions for those showing decreased production of siderophore. Among 6,000 clones screened, nine such clones were selected. These clones contain the transposon inserted in VV2_0830 (GenBank accession number) that is a homolog of a nonribosomal peptide synthase (NRPS). There is an another NRPS module, VV2_0831, 49-bp upstream to VV2_0830. We named these two genes vvs (Vibrio vulnificus siderophore synthase) A and B, respectively. Mutation of either vvsA or vvsB showed a decreased production of siderophore. The expression of an NRPS-lux fusion was negatively modulated by the presence of iron, and the regulation was dependent on Fur (ferric uptake regulator). However, the expression of the NRPS genes was still not fully derepressed in the iron-rich condition, even in furnull mutant cells, suggesting that some other unknown factors are involved in the regulation of the genes. We also demonstrated that the NRPS genes are important for virulence of the pathogen in a mice model.

Fermentation-Mediated Enhancement of Ginseng's Anti-Allergic Activity against IgE-Mediated Passive Cutaneous Anaphylaxis In Vivo and In Vitro

  • Hwang, Seon-Weon;Sun, Xiao;Han, Jun-Hyuk;Kim, Tae-Yeon;Koppula, Sushruta;Kang, Tae-Bong;Hwang, Jae-Kwan;Lee, Kwang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1626-1634
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    • 2018
  • Ginseng (the root of Panax ginseng Meyer) fermented by Lactobacillus plantarum has been found to attenuate allergic responses in in vitro and in vivo experimental models. Ginseng has been reported to also possess various biological functions including anti-inflammatory activity. The present study was aimed at comparing the anti-allergic effect of ginseng and fermented ginseng extracts on IgE-mediated passive cutaneous anaphylaxis in vitro in a murine cell line and in vivo in mice. Fermented ginseng extract (FPG) showed higher inhibitory effect against in vitro and in vivo allergic responses when compared with ginseng extract (PG). The secretion of ${\beta}$-hexosaminidase and interleukin (IL)-4 from the IgE-DNP-stimulated RBH-2H3 mast cells were significantly (p < 0.05) inhibited by FPG treatment, and this effect was concentration-dependent. Further, MKK4 activation and subsequent JNK phosphorylation were attenuated by FPG treatment. The inhibitory effect of FPG on the in vitro allergic response was verified in vivo against IgE-DNP-induced passive cutaneous anaphylaxis in a mouse model. These data indicated that the fermentation of ginseng with L. plantarum enhanced its anti-allergic effects both in vitro and in vivo. We predict that compositional changes in the ginsenosides caused by the fermentation may contribute to the change in the anti-allergic effects of ginseng. The results of our study highlight the potential of the use of FPG as a potential anti-allergic agent.

Effects of Poly-N-acetyl Glucosamine(pGlcNAc) Patch on Wound Healing in db/db Mouse (Poly-N-acetyl-glucosamine이 당뇨병 쥐에서 창상치료에 미치는 영향)

  • Yang, Ho Jik;Yoon, Chi Sun
    • Archives of Plastic Surgery
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    • v.35 no.2
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    • pp.121-126
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    • 2008
  • Purpose: Poly-N-acetyl glucosamine(PGlcNAc) nanofiber-based materials, produced by a marine microalga, have been characterized as effective hemostatic and angiogenic agents. The similarity between PGlcNAc patch and the natural extracellular matrix allows it to support new healthy tissue growth in an injured area and to encourage fluid absorption. In this study, we hypothesized that a poly-N-acetyl glucosamine fiber patch(PGlcNAc patch) may enhance wound healing in the db/db mouse. Methods: PGlcNAc patches were applied on one square centimeter, full-thickness, skin wounds in the db/db mouse model. Wounds(n=15 per group) were dressed with a PGlcNAc nanofiber patch for 1 hour(1 h), 24 hours(24 h) or left untreated(NT). After the application time, patches were removed and wounds were allowed to heal spontaneously. The rate of wound closure was evaluated by digital analysis of unclosed wound area in course of time. At day 10, wounds(n=7 per group) were harvested and quantified with immunohistochemical markers of proliferation(Ki-67) and vascularization (platelet endothelial cell adhesion molecule, PECAM-1). Results: Wounds dressed with PGlcNAc patches for 1 hour closed faster than control wounds, reaching 90% closure in 16.6 days, nine days faster than untreated wounds. Granulation tissue showed higher levels of proliferation and vascularization following 1 h treatment than the 24 h and NT groups. In addition to its hemostatic properties, the PGlcNAc material also appears to accelerate wound closure in healing-impaired genetically diabetic mice. Conclusion: This material, with its combination of hemostatic and wound healing properties, has the potential to be effective agent for the treatment of complicated wounds.

Development of a Magnetoencephalograph System for Small Animals (소동물용 뇌자도 측정 시스템 개발)

  • Kim, J.E.;Kim, I.S.;Kang, C.S.;Kwon, H.;Kim, J.M.;Lee, Y.H.;Kim, K.
    • Progress in Superconductivity
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    • v.13 no.1
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    • pp.18-23
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    • 2011
  • We developed a four-channel first order gradiometer system to measure magnetoencephalogram for mice. We used double relaxation oscillation SQUID (DROS). The diameter of the pickup coil is 4 mm and the distance between the coils is 5 mm. Coil distance was designed to have good spatial resolution for a small mouse brain. We evaluated the current dipole localization confidence region for a mouse brain, using the spherical conductor model. The white noise of the measurement system was about 30 fT/$Hz^{1/2}$/cm when measured in a magnetically shielded room. We measured magnetic signal from a phantom having the same size of a mouse brain, which was filled with 0.9% saline solution. The results suggest that the developed system has a feasibility to study the functions of brain of small animals.

The Effects of Hominis Placenta Hydrate (HPH) on the Bone Marrow Failuer Induced by 5-Fluorouracil

  • Shin, Jang-Woo;Cho, Jung-Hyo;Son, Chang-Gue;Lee, Yong-Yeon;Lee, Yeon-Weol;Yoo, Hwa-Seung;Lee, Nam-Heon;Yun, Dam-Hee;Cho, Chong-Kwan;Choi, Chang-Woo
    • The Journal of Internal Korean Medicine
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    • v.27 no.2
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    • pp.327-335
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    • 2006
  • Aim: To examine the efficacy of Hominis Placenta Hydrate (HPH) on the hemopoiesis in a myelosuppression model system. Methods: Mice were injected with 5-Fluorouracil (5-FU) intraperitoneally and were administered with 200 mg/kg and 1000 mg/kg of HPH. Peripheral blood was analyzed at 5, 9, and 13 days. Histopathologic examination of bone marrow was performed at 5 days after 5-FU injection. The expression of cytokine involved in hemopoiesis was examined by using ELISA kit. Results: The hematology data demonstrated that the treatment of all the agents augmented monocytes and leucocytes counts in the peripheral blood WBC and platelet in HPH treated group were significant increased compared with control group. Also, cell numbers of RBC and Hb were restored. In HPH treated group, expression of IL-3, GM-CSF was significant increased But not TPO. Conclusions: Based on the above results, it is suggested that Hominis Placenta Hydrate is an effective remedy for the bone marrow failure and myelosuppression occurring during chemotherapy.

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Synergic Effect of Trimebutine Combined with Mosapride on Gastrointestinal Dysfunction and Visceral Pain Induced in Stress Models

  • Park, Young-Joon;Park, Yong-Sul;Chung, Zoo-Chul;Nam, Yun-Sung;Chung, Yoon-Hee;Cho, Kwan-Hyung;Choi, Sung-Up;Sohn, Uy-Dong;Park, Eon-Sub;Je, Hyun-Dong;Lee, Choong-Ho;Lee, Moo-Yeol;Jeong, Ji-Hoon
    • Biomolecules & Therapeutics
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    • v.19 no.1
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    • pp.84-89
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    • 2011
  • The present study was undertaken to determine whether combined treatment with prokinetic trimebutine and mosapride has a synergic effect on gastrointestinal motility and visceral pain associated with gastrointestinal dysfunction. To develop effective gastroprokinetic agents with greater potencies than trimebutine or mosapride for the treatment of gastrointestinal tract disease, a mixture of trimebutine and mosapride was designed and prepared. In the present study, treatment with trimebutine alone showed a dose-dependent effect on propelling movements of normal small and large intestine in mice, whereas mosapride effected only small intestine motility. Co-administration of trimebutine with mosapride, a well-established prokinetic drug, produced a synergistic influence on normal small intestine motility, but demonstrated an unclear effect on large intestine motility, with a slight tendency to reduce the propelling time. In a stress model, the small and large intestine motilities were significantly decreased. The reduction of intestine motility was restored to a normal level and the restoring effect was more pronounced in the combined treatment with trimebutine plus mosapride than treatment with trimebutine or mosapride alone. Furthermore, treatment with trimebutine plus mosapride significantly decreased acute visceral pain which was not controlled by trimebutine or mosapride alone. These data suggest that combination therapy with trimebutine plus mosapride has a synergic effect on small and large intestine motility and visceral pain control in gastrointestinal disorders.

Curcumin Inhibits the Activation of Immunoglobulin E-Mediated Mast Cells and Passive Systemic Anaphylaxis in Mice by Reducing Serum Eicosanoid and Histamine Levels

  • Li, Xian;Lu, Yue;Jin, Ye;Son, Jong-Keun;Lee, Seung Ho;Chang, Hyeun Wook
    • Biomolecules & Therapeutics
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    • v.22 no.1
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    • pp.27-34
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    • 2014
  • Curcumin is naturally occurring polyphenolic compound found in turmeric and has many pharmacological activities. The present study was undertaken to evaluate anti-allergic inflammatory activity of curcumin, and to investigate its inhibitory mechanisms in immunoglobulin E (IgE)/Ag-induced mouse bone marrow-derived mast cells (BMMCs) and in a mouse model of IgE/Ag-mediated passive systemic anaphylaxis (PSA). Curcumin inhibited cyclooxygenase-2 (COX-2) dependent prostaglandin $D_2$ ($PGD_2$) and 5-lipoxygenase (5-LO) dependent leukotriene $C_4$ ($LTC_4$) generation dose-dependently in BMMCs. To probe the mechanism involved, we assessed the effects of curcumin on the phosphorylation of Syk and its downstream signal molecules. Curcumin inhibited intracellular $Ca^{2+}$ influx via phospholipase $C{\gamma}1$ ($PLC{\gamma}1$) activation and the phosphorylation of mitogen-activated protein kinases (MAPKs) and the nuclear factor-${\kappa}B$ (NF-${\kappa}B$) pathway. Furthermore, the oral administration of curcumin significantly attenuated IgE/Ag-induced PSA, as determined by serum $LTC_4$, $PGD_2$, and histamine levels. Taken together, this study shows that curcumin offers a basis for drug development for the treatment of allergic inflammatory diseases.

Inhibition of Experimental Systemic Inflammation (Septic Inflammation) and Chronic Bronchitis by New Phytoformula BL Containing Broussonetia papyrifera and Lonicera japonica

  • Ko, Hyun Jeong;Kwon, Oh Song;Jin, Jeong Ho;Son, Kun Ho;Kim, Hyun Pyo
    • Biomolecules & Therapeutics
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    • v.21 no.1
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    • pp.66-71
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    • 2013
  • Broussonetia papyrifera and Lonicera japonica have long been used in the treatment of inflammatory disorders in Chinese medicine, especially respiratory inflammation. Previously, a new phytoformula (BL) containing B. papyrifera and L. japonica was found to exert strong anti-inflammatory activity against several animal models of inflammation, especially against an animal model of acute bronchitis. In the present investigation, the effects of BL on animal models of septic inflammation and chronic bronchitis are examined. Against lipopolysaccharide (LPS)-induced septic inflammation in mice, BL (200-400 mg/kg) reduced the induction of some important proinflammatory cytokines. At 1 h after LPS treatment, BL was found to considerably inhibit TNF-${\alpha}$ production when measured by cytokine array. At 3 h after LPS treatment, BL inhibited the induction of several proinflammatory cytokines, including IFN-${\gamma}$ and IL-$1{\beta}$, although dexamethasone, which was used as a reference, showed a higher inhibitory action on these biomarkers. Against chronic bronchitis induced by LPS/elastase instillation in rats for 4 weeks, BL (200-400 mg/kg/day) significantly inhibited cell recruitment in bronchoalveolar lavage fluid. Furthermore, BL considerably reduced lung injury, as revealed by histological observation. Taken together, these results indicate that BL may have a potential to treat systemic septic inflammation as well as chronic bronchitis.

Poncirin Inhibits Osteoclast Differentiation and Bone Loss through Down-Regulation of NFATc1 In Vitro and In Vivo

  • Chun, Kwang-Hoon;Jin, Hyun Chul;Kang, Ki Sung;Chang, Tong-Shin;Hwang, Gwi Seo
    • Biomolecules & Therapeutics
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    • v.28 no.4
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    • pp.337-343
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    • 2020
  • Activation of osteoclast and inactivation of osteoblast result in loss of bone mass with bone resorption, leading to the pathological progression of osteoporosis. The receptor activator of NF-κB ligand (RANKL) is a member of the TNF superfamily, and is a key mediator of osteoclast differentiation. A flavanone glycoside isolated from the fruit of Poncirus trifoliata, poncirin has anti-allergic, hypocholesterolemic, anti-inflammatory and anti-platelet activities. The present study investigates the effect of poncirin on osteoclast differentiation of RANKL-stimulated RAW264.7 cells. We observed reduced formation of RANKL-stimulated TRAP-positive multinucleated cells (a morphological feature of osteoclasts) after poncirin exposure. Real-time qPCR analysis showed suppression of the RANKL-mediated induction of key osteoclastogenic molecules such as NFATc1, TRAP, c-Fos, MMP9 and cathepsin K after poncirin treatment. Poncirin also inhibited the RANKL-mediated activation of NF-κB and, notably, JNK, without changes in ERK and p38 expression in RAW264.7 cells. Furthermore, we assessed the in vivo efficacy of poncirin in the lipopolysaccharide (LPS)-induced bone erosion model. Evaluating the micro-CT of femurs revealed that bone erosion in poncirin treated mice was markedly attenuated. Our results indicate that poncirin exerts anti-osteoclastic effects in vitro and in vivo by suppressing osteoclast differentiation. We believe that poncirin is a promising candidate for inflammatory bone loss therapeutics.