• BMB Reports
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• v.43 no.1
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• pp.57-61
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• 2010

Activator protein-1 can induce either cell survival or death, which is controlled by opposing effects of different Jun members. It is generally accepted that c-Jun is pro-apoptotic, but that JunD is anti-apoptotic in stress-exposed cells. Additionally, although there are reports suggesting that JunB plays a protective role, its role in stress-induced apoptosis remains unclear. Here, we investigated the role of JunB in $H_2O_2$-induced cell death using cells that over-expressed the protein or were transfected with si-JunB. Inhibition of JunB expression accelerated $H_2O_2$-mediated loss of mitochondrial membrane potential (MMP) and cytotoxicity. Conversely, over-expression of JunB protein led to significant inhibition of the MMP loss and cell death. The increase in JunB expression also attenuated nuclear relocation of apoptosis-inducing factor and mitochondrial Bcl-2 reduction that occurred following $H_2O_2$ exposure. These results suggest that JunB can signal survival against oxidant-mediated cell death by suppressing mitochondrial stress.

• BMB Reports
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• v.33 no.6
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• pp.483-492
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• 2000

The Herpes simplex virus type 1 (HSV-1) strain F glycoprotein H (gH) gene in the pHLB-4 plasmid was recombinated into a baculovirus expression vector (lacZ-HcNPV) to construct a recombinant virus GH-HcNPV expressing gH. The sequences of gH and its expression were analyzed. The gH gene was located in the 6.41 kb BglII fragment. The open reading frame (ORF) of the gH gene was 2,517 bp and codes 838 amino acid residues. Insect cells infected with this recombinant virus synthesized a high level of the matured and gX-gH fusion protein with approximately 112 kDa. The fusion gH protein was localized on the membrane of the insect cells as seen by using immunofluorescence assay and accumulated in the cultured media by the SDS-PAGE and immunoprecipitation assays. The amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. Antibodies raised in mice to this recombinant protein recognized viral gH and neutralized the infectivity of HSV-1 in vitro. These results demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV; accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

• Biomolecules & Therapeutics
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• v.27 no.3
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• pp.290-301
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• 2019

Paeonol has neuroprotective function, which could be useful for improving central nervous system disorder. The purpose of this study was to characterize the functional mechanism involved in brain transport of paeonol through blood-brain barrier (BBB). Brain transport of paeonol was characterized by internal carotid artery perfusion (ICAP), carotid artery single injection technique (brain uptake index, BUI) and intravenous (IV) injection technique in vivo. The transport mechanism of paeonol was examined using conditionally immortalized rat brain capillary endothelial cell line (TR-BBB) as an in vitro model of BBB. Brain volume of distribution (VD) of [$^3H$]paeonol in rat brain was about 6-fold higher than that of [$^{14}C$]sucrose, the vascular space marker of BBB. The uptake of [$^3H$]paeonol was concentration-dependent. Brain volume of distribution of paeonol and BUI as in vivo and inhibition of analog as in vitro studies presented significant reduction effect in the presence of unlabeled lipophilic compounds such as paeonol, imperatorin, diphenhydramine, pyrilamine, tramadol and ALC during the uptake of [$^3H$]paeonol. In addition, the uptake significantly decreased and increased at the acidic and alkaline pH in both extracellular and intracellular study, respectively. In the presence of metabolic inhibitor, the uptake reduced significantly but not affected by sodium free or membrane potential disruption. Similarly, paeonol uptake was not affected on OCTN2 or rPMAT siRNA transfection BBB cells. Interestingly. Paeonol is actively transported from the blood to brain across the BBB by a carrier mediated transporter system.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.50 no.2
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• pp.94-102
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• 2024

The exact mechanism of sialolith formation has yet to be determined. Recurrence of sialolithiasis is rare, affecting only 1%-10% of patients. The current study presents a case of recurrent stones that occurred twice on the right submandibular gland 6 months postoperative and 7 months after reoperation in a 48-year-old female patient. The stones were analyzed using histology, scanning electron microscopy, energy dispersive spectroscopy, and transmission electron microscopy (TEM). The first stone showed a three-layered structure with a poorly mineralized peripheral multilayered zone, highly mineralized middle layer, and the central nidus. The stones were composed of Ca, C, O, Cu, F, N, P, Si, Zn, and Zr. In TEM, compact bi-layered bacterial cell membrane was found on the peripheral layer and the central nidus of the stone as well as exosomes in the central nidus. The results demonstrated the essential components of sialolith formation, including bacteria, inflammatory exosomes, and exfoliated salivary epithelial cells that cooperatively underwent the pathogenetic progresses of central nidus formation, induction of compact zone calcification of the middle layer, and repeated subsequent deposition in the peripheral multilayer zone. The rapid recurrence could have resulted from residual pieces of a sialolith acting as the nidus of bacterial infection.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.392-402
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• 2017

Background: Previously, we reported that Korean Red Ginseng inhibited liver fibrosis in mice and reduced the expressions of fibrogenic genes in hepatic stellate cells (HSCs). The present study was undertaken to identify the major ginsenoside responsible for reducing the numbers of HSCs and the underlying mechanism involved. Methods: Using LX-2 cells (a human immortalized HSC line) and primary activated HSCs, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assays were conducted to examine the cytotoxic effects of ginsenosides. $H_2O_2$ productions, glutathione contents, lactate dehydrogenase activities, mitochondrial membrane permeabilities, apoptotic cell subpopulations, caspase-3/-7 activities, transferase dUTP nick end labeling (TUNEL) staining, and immunoblot analysis were performed to elucidate the molecular mechanism responsible for ginsenoside-mediated cytotoxicity. Involvement of the AMP-activated protein kinase (AMPK)-related signaling pathway was examined using a chemical inhibitor and small interfering RNA (siRNA) transfection. Results and conclusion: Of the 11 ginsenosides tested, 20S-protopanaxadiol (PPD) showed the most potent cytotoxic activity in both LX-2 cells and primary activated HSCs. Oxidative stress-mediated apoptosis induced by 20S-PPD was blocked by N-acetyl-$\text\tiny L$-cysteine pretreatment. In addition, 20S-PPD concentration-dependently increased the phosphorylation of AMPK, and compound C prevented 20S-PPD-induced cytotoxicity and mitochondrial dysfunction. Moreover, 20S-PPD increased the phosphorylation of liver kinase B1 (LKB1), an upstream kinase of AMPK. Likewise, transfection of LX-2 cells with LKB1 siRNA reduced the cytotoxic effect of 20S-PPD. Thus, 20S-PPD appears to induce HSC apoptosis by activating LKB1-AMPK and to be a therapeutic candidate for the prevention or treatment of liver fibrosis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.10
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• pp.1599-1605
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• 2005

The object of this study is to develop the method for differentiating fresh meat from frozen meat by using the measurement of the mitochondrial malate dehydrogenase in the Korean native cattle. The principle of this experiment is based on the fact that the enzyme proteins associated with mitochondrial membrane could be released by freezing. The methods of differentiating fresh meat from thawed, frozen meat were studied by measurements of mitochondrial malate dehydrogenase activity of meat press juice. Fresh and frozen beef were stored at 4, -4, -18 and -77$^{\circ}C$ for 15-day storage period. A meat press machine using air pressure was manufactured especially for these experiments, and sufficient amount of drip (about 0.15 mL/g) from 1.5 g of beef sample was efficiently obtained under a pressure of 8 kg/$cm^{2}$ generated by the meat pressing machine. The mitochondrial malate dehydrogenase activities of frozen meat drip i년ices stored at -18 and -77$^{\circ}C$ were significantly higher than those of fresh and frozen meat samples at -4$^{\circ}C$ (p < 0.05) during 10-min reaction period. However, the enzyme activities of the frozen meat drip juices (-18 and -77$^{\circ}C$) disappeared after 5 minutes of the reaction, which was not observed from the fresh and -4$^{\circ}C$ frozen meats. The enzyme activity maintained until 12 minutes for the fresh and -4$^{\circ}C$ frozen meats. From these results, the mitochondrial malate dehydrogenase could be considered as an indicator to differentiate fresh beef from frozen one.

• Korean Journal of Ichthyology
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• v.27 no.2
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• pp.63-70
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• 2015

This research is about spawning behavior and morphological development of the eggs and larvae of fringed blenny which lives in Korean sea. It helps to find out the relation between fringed blenny larvae and other stichaeidae larva. Moreover, it is going to use for the basic data of taxonomical research. There were 8 mother fishes (total length 31.8~34.0 cm, average $32.9{\pm}0.21cm$), which were captured from the cost of Sindeok-dong, Yeosu-si, Jeollanam-do from December, 2010. we used trap to capture them. Mother fishes were brought to glass aquarium ($50{\times}30{\times}35cm$) and nurtured with recirculate rearing system. Breeding water temperature was $12.3{\sim}15.8^{\circ}C$ (average $14.1{\pm}2.47$) and salinity was 32.5~33.5‰ (average $33.0{\pm}0.05$‰). fertilized egg was round, opaque demersal egg. The size was 1.81~2.19 mm (average $2.00{\pm}0.27mm$, n=50). Incubation was begun after 271 hours and 30 minutes from fertilization. It started with piercing the egg membrane. prelarvae had length of 8.45~8.84 mm (average $8.65{\pm}0.28mm$, n=5) right after the incubation, and both mouth and anus were opened. After 11 days from the incubation, postlarvae had length of 10.5~11.3 mm (average $10.9{\pm}0.57mm$), and the number of myotomes was increased to 66.

• Journal of Life Science
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• v.26 no.11
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• pp.1232-1236
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• 2016

Mitochondrial calcium uptake 1 (MICU1) including two canonical EF hands, located in the mitochondrial inner membrane, is known to play a crucial role in the calcium uptake in mitochondria. Mitochondrial calcium uptake in muscular cells is related to post mortem shortening by calcium release from muscles. Therefore, the porcine MICU1 gene has been estimated as a genetic candidate for meat quality traits. In this study, variations on the exonic regions of the porcine MICU1 gene were investigated by sequencing cDNAs and tested for their association with meat quality traits. A total of 667 Berkshire heads (347 sows and 320 castrated boars) were used for this association test. Three SNPs were detected on the 3' untranslated region (UTR) of the porcine MICU1 gene. SNP1 (c.*136G>A) was associated with drip loss (p=0.017) and intramuscular fat content (p=0.039). In addition, SNP2 (c.*222G>A) and SNP3 (c.*485G>A) were associated with drip loss (p=0.018) and intramuscular fat content (p<0.001), respectively. In conclusion, it was verified that three variations on the 3' UTR of the porcine MICU1 gene were significantly associated with meat quality traits. It was also suggested that molecular biological analyses are needed to validate the function of variations on the 3 UTR of the porcine MICU1 gene.

• Molecules and Cells
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• v.38 no.12
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• pp.1064-1070
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• 2015

Translocator protein 18 kDa (TSPO) is a mitochondrial outer membrane protein and is abundantly expressed in a variety of organ and tissues. To date, the functional role of TSPO on vascular endothelial cell activation has yet to be fully elucidated. In the present study, the phorbol 12-myristate 13-acetate (PMA, 250 nM), an activator of protein kinase C (PKC), was used to induce vascular endothelial activation. Adenoviral TSPO overexpression (10-100 MOI) inhibited PMA-induced vascular cell adhesion molecule-1 (VCAM-1) and intracellular cell adhesion molecule-1 (ICAM-1) expression in a dose dependent manner. PMA-induced VCAM-1 expressions were inhibited by Mito-TEMPO ($0.1-0.5{\mu}m$), a specific mitochondrial antioxidants, and cyclosporin A ($1-5{\mu}m$), a mitochondrial permeability transition pore inhibitor, implying on an important role of mitochondrial reactive oxygen species (ROS) on the endothelial activation. Moreover, adenoviral TSPO overexpression inhibited mitochondrial ROS production and manganese superoxide dismutase expression. On contrasts, gene silencing of TSPO with siRNA increased PMA-induced VCAM-1 expression and mitochondrial ROS production. Midazolam ($1-50{\mu}m$), TSPO ligands, inhibited PMA-induced VCAM-1 and mitochondrial ROS production in endothelial cells. These results suggest that mitochondrial TSPO can inhibit PMA-induced endothelial inflammation via suppression of VCAM-1 and mitochondrial ROS production in endothelial cells.