• BMB Reports
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• v.45 no.1
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• pp.32-37
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• 2012

Alternative splicing generates several interleukin-6 (IL-6) isoforms; for them an antagonistic activity to the wild-type IL-6 has been proposed. In this study we quantified the relative abundance of IL-6 mRNA isoforms in a panel of mouse tissues and in C2C12 cells during myoblast differentiation or after treatment with the $Ca^{2+}$ ionophore A23187, the AMP-mimetic AICAR and TNF-${\alpha}$. The two mouse IL-6 isoforms identified, IL-6${\delta}$5 (deletion of the first 58 bp of exon 5) and IL-6${\delta}$3 (lacking exon 3), were not conserved in rat and human, did not exhibit tissue specific regulation, were expressed at low levels and their abundance closely correlated to that of full-length IL-6. Species-specific features of the IL-6 sequence, such as the presence of competitive 3' acceptor site in exon 5 and insertion of retrotransposable elements in intron 3, could explain the production of IL-6${\delta}$5 and IL-6${\delta}$3. Our results argued against biological significance for mouse IL-6 isoforms.

• Applied Microscopy
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• v.27 no.2
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• pp.145-153
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• 1997

Histopathological and fine structural changes in mouse skin after injection of venoms extracted from the venom glands of the honeybee, Apis mellifera, were studied with light and electron microscopes. At this experiment the venoms were directly injected at the hairless abdominal skin of the mouse through the sting of the bee's venomous organ. Main changes appeared within one hour after injection at both epithelial and connective tissues as considerable hyperemia and angioedema, and slight edema and fibrosis. High magnified electron micrographs reveal not only increase of diameter but also deposition of electron dense grains (which seems to be an auto immunoglobulin) at the collagenous fibers characteristically. This kinds of histological and fine structural responses were diminished from 12 hour after injection, and the pathological symptoms disappeared within 3 days at most cases. So, the skin responses induced by honeybee venom seem to be not severe compare to other cases reported by other venomous arthropods.

• Proceedings of the Korean Vacuum Society Conference
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• 2011.02a
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• pp.20-20
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• 2011

It has been known that, under certain conditions, application of low-temperature atmospheric-pressure plasmas can enhance proliferation of cells. In this study, conditions for optimal cell proliferation were examined for various cells relevant for orthopaedic applications. Plasmas used in our experiments were generated by dielectric barrier discharge (DBD) with a helium flow (of approximately 3 litter/min) into ambient air at atmospheric pressure by a 10 kV~20 kHz power supply. Such plasmas were directly applied to a medium, in which cells of interest were cultured. The cells examined in this study were human synoviocytes, rat mesenchymal stem cells derived from bone marrow or adipose tissue, a mouse osteoblastic cell line (MC3T3-E1), a mouse embryonic mesenchymal cell line (C3H-10T1/2), human osteosarcoma cells (HOS), a mouse myoblast cell line (C2C12), and rat Schwann cells. Since cell proliferation can be enhanced even if the cells are not directly exposed to plasmas but cultured in a medium that is pre-treated by plasma application, it is surmised that long-life free radicals generated in the medium by plasma application stimulate cell proliferation if their densities are appropriate. The level of free radical generation in the medium was examined by dROMs tests and correlation between cell proliferation and oxidative stress was observed. Other applications of plasma medicine in orthopaedics, such as plasma modification of artificial bones and wound healing effects by direct plasma application for mouse models, will be also discussed. The work has been done in collaboration with Prof. H. Yoshikawa and his group members at the School of Medicine, Osaka University.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.28 no.2
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• pp.114-125
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• 2002

This study was designed to evaluate the influence of fibroblasts or connective tissue from mouse oral mucosa on differentiation of neonatal mouse calvaria-derived osteoblasts and mineralization of bone nodules. Primary cell cultures from mouse calvarial osteoblasts and 2-4 passaged fibroblasts from oral mucosa were co-cultured in monolayer cultures, devided into 6 experimental group according to cell density or cell confluency. Osteoblasts were also co-cultured with fibroblasts in $Transwell^{(R)}$ culture plate with different co-cultured period according to osteoblast differentiation. The alkaline phosphatase activity were measured in monolayer cultures and cultures using $Transwell^{(R)}$. The mineralized bone nodules were presented by Von Kossa staining and density of mineralized nodules was measured by image analysis. The connective tissues with or without osteoblast seeding were cultured and examined histologically by Von Kossa and Trichrome Goldner staining. The results were as follows; 1. Prolonged maturation of matrix and delayed mineralization of bone nodules were resulted in monolayer cultures. 2. Co-culture of fibroblast with osteoblast using $Transwell^{(R)}$ during osteoblast proliferation stage stimulated proliferation of osteoblasts and increased alkaline phosphatase activity and mineralization of bone nodules. 3. Co-culture of fibroblast with osteoblast using $Transwell^{(R)}$ during matrix mineralization stage decreased and delayed mineralization of bone nodules. 4. In vitro cultured connective tissue with osteoblast seeding resulted in proliferation of osteoblasts and matrix formation with mineralization.

• BMB Reports
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• v.29 no.6
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• pp.555-563
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• 1996

A membrane-bound phosphatidylinositol 4-kinase (PI 4-kinase) was separated in a sucrose gradient and solubilized with 1% Triton X-100 from mouse brain. The enzyme was purified 2,952-fold by various chromatographic techniques including DEAE-cellulose, PI-Sepharose and Sephacryl S-200 gel filtration. The molecular weight of PI 4-kinase was approximately 76 kDa by gel filtration and 70.8 kDa by SDS-polyacrylamide gel electrophoresis. The purified enzyme exhibited specific activity of 11.2 nmol/min/mg protein and pi value of 4.7. Kinetic analysis of the PI 4-kinase indicated apparent $K_m$, values of 190 ${\mu}M$ and 120 ${\mu}M$ for phosphatidylinositol and ATP, respectively. The maximal activity of this purified enzyme was observed at pH 7.4 at an incubation temperature of $37^{\circ}C$. The enzyme activity was significantly activated by $Mg^{2+}$, $Mn^{2+}$ and $Fe^{2+}$, and inhibited severely by $Ca^{2+}$. PI 4-kinase was proved to be pure in its immunoblot test by polyclonal antibody prepared from immunized rabbit sera. By this test, we were able to detect the existence of the same type of PI 4-kinase from other mouse organ tissues, such as liver, heart, kidney and spleen. Furthermore, similar immunoblot analysis with the same antisera recognized the different epitopes of PI 4-kinase proteins from various organs of rabbit, chinese hamster and rat.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.12
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• pp.1517-1522
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• 2007

Inhibitory effects of allergic contact dermatitis of hot water extract of Duchung (Eucommia ulmoides Oliver) leaf, bark and stem growing at Sancheong-gun were investigated for female BALB/c mouse induced by repeat elicitation of DNCB (2,4-dinitro-chlorobenzene). Skin reactions, consisting of increased ear thickness and the presence of ear inflammation, were observed in mice treated with DNCB and Duchung. Weight of lymph node, spleen and thymus in mice treated with Duchung extracts were lower than that of mouse treated with DNCB. Ear weight of mouse treated with Duchung extracts was decreased by increasing the concentration of sample as compared to control group and dropped as low as control level at 1,000 mg/kg. Ear thickness became thinner as test time on Duchung extract progressed. MDA (malondialdehyde) contents in liver tissue were not different in sample group with DNCB group, but were different in ear tissue. NO (nitric oxide) contents was decreased in Duchung extract groups at serum and ear tissues as compared to 1% DNCB group. In the present study, the results suggested that Duchung extract inhibits inflammatory response and oxidative damage induced DNCB allergen.

• Development and Reproduction
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• v.23 no.3
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• pp.213-221
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• 2019

The epididymal fat of mouse is a part of visceral fat deposit and is divided into the distal or proximal part based on its histochemical characteristics. Even though the formation of the epididymal fat pad begins before the birth, a detailed adipogenic procedure of the epididymal fat has not been revealed. The epididymal fat pad becomes enlarged and expended with age, and expressional changes of numerous genes are associated with the maturation of fat tissues. In the present research, expressional patterns of adipose tissue-related genes in the distal epididymal fat of mouse at 2, 5, 8, and 12 months of postnatal age were determined by a quantitative real-time polymerase chain reaction (PCR) analysis. The lowest transcript levels of fatty acid binding protein 4 (Fabp4), lipoprotein lipase (Lpl), delta like non-canonical Notch ligand 1 (Dlk1), peroxisome proliferator-activated receptor gamma (Pparg), leptin (Lep), adiponectin (Adipoq), and resistin (Retn) were detected at 2 months of age, except fatty acid synthase (Fasn) showing the lowest level at 5 months of age. Even though expression of Lep and Fabp4 were gradually increased until 12 months of age, significant increases of Pparg and Adipoq transcript levels were continued until 8 months of age. The transcript levels of Lpl, Rent, Dlk1, and Fasn were significantly increased at 8 months of age, compared with those at 2 months of age. The current findings suggest that the expansion of the distal epididymal fat of mouse during postnatal period would be companied with differential expression of various adipocyte-associated molecules.

• The Korean Journal of Physiology and Pharmacology
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• v.25 no.1
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• pp.27-38
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• 2021

Excessive salt intake induces hypertension, but several gamma-aminobutyric acid (GABA) supplements have been shown to reduce blood pressure. GABA-salt, a fermented salt by L. brevis BJ20 containing GABA was prepared through the post-fermentation with refined salt and the fermented GABA extract. We evaluated the effect of GABA-salt on hypertension in a high salt, high cholesterol diet induced mouse model. We analyzed type 1 macrophage (M1) polarization, the expression of M1 related cytokines, GABA receptor expression, endothelial cell (EC) dysfunction, vascular smooth muscle cell (VSMC) proliferation, and medial thicknesses in mice model. GABA-salt attenuated diet-induced blood pressure increases, M1 polarization, and TNF-α and inducible nitric oxide synthase (NOS) levels in mouse aortas, and in salt treated macrophages in vitro. Furthermore, GABA-salt induced higher GABAB receptor and endothelial NOS (eNOS) and eNOS phosphorylation levels than those observed in salt treated ECs. In addition, GABA-salt attenuated EC dysfunction by decreasing the levels of adhesion molecules (E-selectin, Intercellular Adhesion Molecule-1 [ICAM-1], vascular cell adhesion molecule-1 [VCAM-1]) and of von Willebrand Factor and reduced EC death. GABA-salt also reduced diet-induced reductions in the levels of eNOS, phosphorylated eNOS, VSMC proliferation and medial thickening in mouse aortic tissues, and attenuated Endothelin-1 levels in salt treated VSMCs. In summary, GABA-salt reduced high salt, high cholesterol diet induced hypertension in our mouse model by reducing M1 polarization, EC dysfunction, and VSMC proliferation.

• BMB Reports
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• v.56 no.5
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• pp.308-313
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• 2023

Phenotypic features such as ataxia and loss of motor function, which are characteristics of Parkinson's disease (PD), are expected to be very closely related to cerebellum function. However, few studies have reported the function of the cerebellum. Since the cerebellum, like the cerebrum, is known to undergo functional and morphological changes due to neuroinflammatory processes, elucidating key functional factors that regulate neuroinflammation in the cerebellum can be a beneficial therapeutic approach. Therefore, we employed PD patients and MPTP-induced PD mouse model to find cytokines involved in cerebellar neuroinflammation in PD and to examine changes in cell function by regulating related genes. Along with the establishment of a PD mouse model, abnormal shapes such as arrangement and number of Purkinje cells in the cerebellum were confirmed based on histological finding, consistent with those of cerebellums of PD patients. As a result of proteome profiling for neuroinflammation using PD mouse cerebellar tissues, fetuin-A, a type of cytokine, was found to be significantly reduced in Purkinje cells. To further elucidate the function of fetuin-A, neurons isolated from cerebellums of embryos (E18) were treated with fetuin-A siRNA. We uncovered that not only the population of neuronal cells, but also their morphological appearances were significantly different. In this study, we found a functional gene called fetuin-A in the PD model's cerebellum, which was closely related to the role of cerebellar Purkinje cells of mouse and human PD. In conclusion, morphological abnormalities of Purkinje cells in PD mice and patients have a close relationship with a decrease of fetuin-A, suggesting that diagnosis and treatment of cerebellar functions of PD patients might be possible through regulation of fetuin-A.

• Proceedings of the Korean Vacuum Society Conference
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• 2015.08a
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• pp.230.1-230.1
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• 2015

One of the major problems of biological ToF-SIMS imaging is the lack of protein and peptide imaging. Most of biological story telling is mianly based on proteins. The biological implication of lipid ToF-SIMS imaging would be much higher if protein imaging is provided together. Utilizing high secondary ion yields of metals, proteins can be ToF-SIMS imaged with nanoparticle tagged proteins. Nanoparticles such as Fe3O4, SiO2, PbS were used for imaing NeuN, MCH, Orexin A, ${\alpha}$ synucline, TH(Tryosine Hydroxylase) in mouse tissues with a spatial resolution of ${\sim}2{\mu}m$ using a TOF-SIMS. Lipids and neurotransmitters images obtained simultaneously with protein images were overlayed for more deeper understanding of neurobiology, which is not allowed by any other bioimaging technqiues. The protein images from TOF-SIMS were compared with confocal fluorescence microscopy and NanoSIMS images. A new sample preparation method for imaging single cell membranes in a tissue using the vibrotome technique to prepare a tissue slice without any fixation and freeze drying will be also presented briefly for Hippocampus and Hypothalamus tissues.