• BMB Reports
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• v.41 no.10
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• pp.745-750
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• 2008

Selenium, an essential trace element possessing anti-carcinogenic properties, can induce apoptosis in cancer cells. We have previously shown that sodium selenite can induce apoptosis by activating the mitochondrial apoptosis pathway in NB4 cells. However, the detailed mechanism remains unclear. Presently, we demonstrate that p53 contributes to apoptosis by directing signaling at the mitochondria. Immunofluorescent and Western blot procedures revealed selenite-induced p53 translocation to mitochondria. Inhibition of p53 blocked accumulation of reactive oxygen species (ROS) and loss of mitochondrial membrane potential, suggesting that mitochondrial p53 acts as an upstream signal of ROS and activates the mitochondrial apoptosis pathway. Selenite also disrupted cellular calcium ion homeostasis in a ROS-dependent manner and increased mitochondrial calcium ion concentration. p38 kinase mediated phosphorylation and mitochondrial translocation of p53. Taken together, these results indicate that p53 involves selenite-induced NB4 cell apoptosis by translocation to mitochondria and activation mitochondrial apoptosis pathway in a transcription-independent manner.

• Reproductive and Developmental Biology
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• v.35 no.4
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• pp.427-434
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• 2011

Mitochondria diseases have been reported to involve structural and functional defects of complex I-V. Especially, many of these diseases are known to be related to dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I). The dysfunction of mitochondria complex I is associated with neurodegenerative disorders, such as Parkinson's disease, Huntington's disease, and Leber's hereditary optic neuropathy (LHON). Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) is largest and consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. The Saccharomyces cerevisiae NDI1 gene using a recombinant adeno-associated virus vector (rAAV-NDI1) was successfully expressed in AML12 mouse liver hepatocytes and the NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced cells was not affected by rotenone which is inhibitor of complex I, but was inhibited by antimycin A. Furthermore, these results indicate that Ndi1 can be functionally expressed in the AML12 mouse liver hepatocytes. It is conceivable that the NDI1 gene is powerful tool for gene therapy of mitochondrial diseases caused by complex I deficiency. In the future, we will attempt to functionally express the NDI1 gene in mouse embryonic stem (mES) cell.

• Korean Journal of Weed Science
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• v.30 no.3
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• pp.225-232
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• 2010

The effect of Protox expression site on herbicidal resistance was investigated in wild-type and transgenic rice plants imposed by peroxidizing herbicide oxyfluorfen. The transgenic rice systems involved the plastidal expression of Arabidopsis protoporphyrinogen oxidase (Protox; AP line) and the dual expression of Myxococcus xanthus Protox in chloroplasts and mitochondria (TTS line). The oxyfluorfen-treated TTS4 line showed the lower levels of cellular leakage and malonyldialdehyde and the sustained capacity of 5-aminolevulinic acid synthesis, compared to the oxyfluorfen-treated AP and wild-type lines. During oxyfluorfen action, the TTS4 line had greater herbicide resistance than the AP1 line, indicating that the dual expression of M. xanthus Protox in chloroplasts and mitochondria prevented the accumulation of photodynamic protoporphyrin IX more effectively than the expression of Arabidopsis Protox only in chloroplasts. These results suggest that the ectopic expression of Protox in mitochondria greatly contributes to the herbicidal resistance in rice plants.

• Journal of Life Science
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• v.10 no.4
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• pp.340-346
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• 2000

This study was designed to investigate the effects of silk fibroin(Mw 500) powder (SFP) on oxygen radicals and the scavenger enzymes in brain membranes of rats. Spragu-Dawley(SD) male rats(160${\pm}$10g) were fed basic diet(control group), and experimental diets(SFP-2.5 and SFP-5.0 groups) added 2.5 and 5.0g/kg BW/day for 6 weeks. Hydroxyl radical($.$OH) levels resulted in a decreases(6.6% and 9.7%, 2.8% and 11.9%, respectively) in brain mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group, but were significantly decreased in these membrances of SFP-5.0 group only. Superoxide radical (O2) levels were a slightly decreased (2.0% and 9.1%, respectively) in brain cytosol of SFP-2.5 and SFP-5.0 groups compared with control group. Lipid peroxide(LPO) levels were significantly decreased (12.9% and 21.9%, 13.2% and 22.5%, respectively) in brain mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. Oxidized protein (OP) levels were significantly decreased (16.7% and 15.7%, respectively) in brain microsomes of SFP-2.5 and SFP-5.0 group compared with control group, but significantly difference between in brain mitochondria of these two groups could not be obtained. Mn-SOD activities were remarkably increased (11.2% and 24.2%, respectively) in mitochodria of SFP-2.5 and SFP-5.0 groups. CuZn-SOD activities were effectively increased (7.7% and 19.6%, respectively) in brain cytosol of SFP-2.5 and SFP-5.0 groups, but significant difference between control and SFP-2.5 groups could be not obtained. GSHPx activities were considerably increased (5.3% and 11.7%, respectively) in brain cytosol of SFP-2.0 and SFP-5.0 groups compared with control group. There results suggest that anti-aging effect of silk fibroin may play an effective learning and memory role in a attenuating a oxidative stress and increasing a scavenger enzyme activity in brain membranes.

• Journal of Life Science
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• v.10 no.6
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• pp.577-583
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• 2000

This study was designed to investigate the effect of silk fibroin (Mw 500)power (SFP) on oxygen radicals and their scavenger enzymes in liver membranes of rats. Sprague-Dawley (SD) male rats (160$\pm$10 g) were fed basic diet (control group), and experimental diets (SFP-2.5 and SFP-5.0 groups) added 2.5 and 5.0 g/kg BW/day for 6 weeks. Hydroxyl radical (.OH) levels resulted in a considerable decreases (5.8% and 8.4%, 3.7% and 11.1%, respectively) in liver motochondria and micorsomes of SFP-2.5 and SFP-5.0 groups compared with control group, and $O_2$radical level was remarkably decreased about 15% and 20% in liver cytosol of SFP-2.5 and SFP-5.0 groups compared with control group. Lipid peroxide (LPO) levels were significantly decreased (8.3% and 18.0%, 13.4% and 18.4%, respectively) in liver mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. Oxidized protein (OP) levels were remarkably decreased about 11.6% in liver mitochondria of SFP-5.0 group compared with control group. Mn-SOD activities were remarkably in creased (17.6% and 28.8%, respectively) in mitochondria of SFP-2.5 and SFP-5.0 groups, and Cu/Zn-SOD activities were also effectively in creased (about 14.4%) in liver cytosol of SFP-5.0 groups, but significant difference between GSHPx activity in liver cytosol of these two groups could be not obtained. These results suggest that anti-aging effect of silk fibroin may play an effective anti-aging role in a aattenuating a oxidative stress and increasing a scavenger enzyme activity in liver membranes.

• Toxicological Research
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• v.17 no.3
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• pp.215-223
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• 2001

Cadmium is an ubiquitous toxic metal and chronic exposure to cadmium results in the accumulation of cadmium in the liver and kidneys. In contrast, acute exposure leads to damage mainly in the liver. Apoptosis induced by cadmium has been shown in many tissues in vivo and in cultured cells in vitro. However, the molecular mechanism of cadmium-induced apoptosis is not clear in hepatocyte. To investigate the induction of apoptosis in the hepatocyte, we used mouse hepatoma cell line, Hepalclc7 cells, and analysed the molecules that involved in cadmium-induced apoptosis. Cadmium induced the genomic DNA fragmentation, PARP cleavage, and activation of caspase-3 like protease. Caspase-9 cysteine protease was activated in a time-dependent manner but caspase-8 cysteine protease was not significantly activated in cadmium-treated Hepalclc7 cells. Cadmium also induced mitochondrial dysfunction including cytochrome c release from mitochondria, change oj mitochondrial membrane potential tranition, and tranlocation of Bax Protein into mitochondria. These results strong1y indicated that the signal Pathway of apoptotic death in cadmium-treated Hepalclc7 cells is modulated by caspase cascade via mitochondria.

• Korean journal of applied entomology
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• v.38 no.2
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• pp.129-138
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• 1999

This study was carried out with two types of silkworm, Bombyx mori, adults having different life span: long life span (adult living for more than fifteen days; LLS), and short life span (adult living less for than five days; SLS). The fine structures of fat body cells in the young and the senescent males and females were compared in an attempt to elucidate the aging physiology of the silkworm adult. In SLS, there were many rough endoplasmic reticula (RER) and glycogen granules in the female cytoplasm, whereas the smooth endoplasmic reticula (SER) were observed in the male cytoplasm. Also in SLS, the mitochondria were swollen and had many poor osmiophillic inclusions (fatty degeneration) at three days after emergence. In LLS, compared to SLS, comparatively normal mitochondria and nuclear membrane were observed in even five day old female. In fifteen day old LLS female, however most of cell membranes were disappeared and mitochondria were abnormally expanded. Many fat granules were observed in the cytoplasm of ten day old male (LLS) which died with full autophagic vacuole (AV) collapsed at fifteen days. Therefore, it is suggested that SLS type is recommended for the study of aging tissue, because it is easier to monitor the changes in tissue structure.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.357-366
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• 1996

The present experiments aimed to investigate the metabolism of calcium during oocyte maturation in rat. The concentration of free calcium and calmodulin in oocytes was measured respectively by using of fluo-3/AM and FITC with microscope fluorescence spectrometer. The ultrastructural localization of calcium precipitates in oocytes was observed with the transmission electron microscope. Cumulus-free immature oocytes(GV-oocyte) were cultured in vitro through 15 hours. The free calcium concentration in GV oocyte was $55.9{\pm}3.5nM$. In calcium-containing medium, the free calcium concentration was increased in germinal vesicle breakdown(GVBD) oocyte($64.2{\pm}7.3nM$). In normal medium after calcium chelator treatment ($10{\mu}M$ BAPTA/AM), the free calcium contents were slightly lower than those in control group. In calcium-free medium, the free calcium content was drastically increased in GVBD($72.7{\pm}3.4nM$) and metaphase I - anaphase I ($88.0{\pm}3.4nM$) oocyte. In maturation rate of oocytes, GVBD rate was high in control group($82.9{\pm}6.55%$) and calcium chelator treatment group($91.2{\pm}4.4%$), but in calcium-free medium group, it was low and then the oocyte was degenerated without polar body formation. Relative content of calmodulin in oocyte was significantly(P<0.001) increased in metaphase I - anaphase I than in GV and GVBD oocyte. The calcium precipitates were observed in mitochondria and cytoplasm of GV oocyte but that were not observed in mitochondria of GVBD and metaphase I - anaphase I oocyte. And then the calcium precipitates reappeared in mitochondria of metaphase II oocyte. The above results indicate that changes in free calcium and calmodulin concentration of oocyte occur according to the maturational stages and the extracellular calcium is required during oocyte maturation. Also change of calcium localization in oocyte occurs according to the maturational stages.

• The Korean Journal of Malacology
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• v.27 no.1
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• pp.55-65
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• 2011

Some characteristics of germ cell differntiations during spermiogenesis and mature sperm ultrastructure in male Phacosoma japonicus were investigated by transmission electron microscope observations. The morphology of the spermatozoon of this species has a primitive type and is similar to those of other species in the subclass Heterodonta. Morphologies of the sperm nucleus and the acrosome of this species are the cylindrical type and cap shape, respectively. The spermatozoon is approximately 45-50 ${\mu}m$ in length, including a long curved sperm nucleus (about $3.70{\mu}m$ long with 45 $^{\circ}$ of the angle of the nucleus, an acrosome (about $0.55{\mu}m$ in length), and tail flagellum (about 42-$47{\mu}m$)The axoneme of the sperm tail shows a 9+2 structure. As some characteristics of the acrosomal vesicle structures, the basal and lateral parts of basal rings show electron opaque part (region), while the anterior apex part of the acrosomal vesicle shows electron lucent part (region). These characteristics of the acrosomal vesicle were found in the family Veneridae and other several families in the subclass Heterodonta. These common characteristics of the acrosomal vesicle in the subclass Heterodonta can be used for phylogenetic and systematic analysis as a taxonomic key or a significant tool. The number of mitochondria in the sperm midpiece of this species are four, as one of common characteristics appear in most species in the family Veneridae and other families in the subclass Heterodonta. However, exceptionally, only three species in Veneridae of the subclass Heterodonta contain 5 mitochondria. The number of mitochondria in the sperm midpiece can be used for the taxonomic analysis of the family or superfamily levels as a systematic key or tools.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.6
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• pp.599-607
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• 2017

Most normal cells express L-type amino acid transporter 2 (LAT2). However, L-type amino acid transporter 1 (LAT1) is highly expressed in many tumor cells and presumed to support their increased growth and proliferation. This study examined the effects of JPH203, a selective LAT1 inhibitor, on cell growth and its mechanism for cell death in Saos2 human osteosarcoma cells. FOB human osteoblastic cells and Saos2 cells expressed LAT1 and LAT2 together with their associating protein 4F2 heavy chain, but the expression of LAT2 in the Saos2 cells was especially weak. JPH203 and BCH, a non-selective L-type amino acid transporter inhibitor, potently inhibited L-leucine uptake in Saos2 cells. As expected, the intrinsic ability of JPH203 to inhibit L-leucine uptake was far more efficient than that of BCH in Saos2 cells. Likewise, JPH203 and BCH inhibited Saos2 cell growth with JPH203 being superior to BCH in this regard. Furthermore, JPH203 increased apoptosis rates and formed DNA ladder in Saos2 cells. Moreover, JPH203 activated the mitochondria-dependent apoptotic signaling pathway by upregulating pro-apoptotic factors, such as Bad, Bax, and Bak, and the active form of caspase-9, and downregulating anti-apoptotic factors, such as Bcl-2 and Bcl-xL. These results suggest that the inhibition of LAT1 activity via JPH203, which may act as a potential novel anti-cancer agent, leads to apoptosis mediated by the mitochondria-dependent intrinsic apoptotic signaling pathway by inducing the intracellular depletion of neutral amino acids essential for cell growth in Saos2 human osteosarcoma cells.