• Title/Summary/Keyword: mitochondria

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Haplotype Diversity and Gene Flow of the Diamondback Moth, Plutella xylostella(L.) (Lepidoptera: Yponomeutidae), in Korea (배추좀나방(나비목: 집나방과)의 haplotype 다양성과 유전자 이동률)

  • 김익수;배진식;최광호;진병래;이경로;손흥대
    • Korean journal of applied entomology
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    • v.39 no.1
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    • pp.43-52
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    • 2000
  • A portion of mitochondria1 COI gene (438 bp) was sequenced from the sampls of Plutella xylostella from four localities in Korea to investigate the population genetic structure and characteristics by measuring the magnitude of genetic diversity and the degree of gene flow among populations. Thirteen haplotypes ranging in nucleotide divergence 0.3% to 1.4%, were obtained from 21 individuals. The nucleotide divergence was similar to the other related studies, but haplotype diversity was substantially higher (mean h = 0.81). The genetic distance among geographically remote Cheju Island population and the two Kimhae populations, distant 1 lkm to each other, was not statistically significant (p<0.05). Instead, a substantial or high female gene flow was detected (Nm = 2-30). One Hawaiian haplotype of the diamondback moth obtained through GenBank search also was genetically similar to the ones obtained from this study. Collectively, the genetic population structure of the diamondback moth in Korea can be characterized into two aspects. First, the diamondback moths in Korea possesses overall moderate genetic divergence based on a high number of haplotypes. Second, a high haplotype diversity within each population due to the long distance dispersal with a substantial dispersal power and the resultant genetic similarity among geographic populations is characteristic.

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Mitochondrial Damage and Metabolic Compensatory Mechanisms Induced by Hyperoxia in the U-937 Cell Line

  • Scatena, Roberto;Messana, Irene;Martorana, Giuseppe Ettore;Gozzo, Maria Luisa;Lippa, Silvio;Maccaglia, Alessandro;Bottoni, Patrizia;Vincenzoni, Federica;Nocca, Giuseppina;Castagnola, Massimo;Giardina, Bruno
    • BMB Reports
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    • v.37 no.4
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    • pp.454-459
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    • 2004
  • Experimental hyperoxia represents a suitable in vitro model to study some pathogenic mechanisms related to oxidative stress. Moreover, it allows the investigation of the molecular pathophysiology underlying oxygen therapy and toxicity. In this study, a modified experimental set up was adopted to accomplish a model of moderate hyperoxia (50% $O_2$, 96 h culture) to induce oxidative stress in the human leukemia cell line, U-937. Spectrophotometric measurements of mitochondrial respiratory enzyme activities, NMR spectroscopy of culture media, determination of antioxidant enzyme activities, and cell proliferation and differentiation assays were performed. The data showed that moderate hyperoxia in this myeloid cell line causes: i) intriguing alterations in the mitochondrial activities at the levels of succinate dehydrogenase and succinate-cytochrome c reductase; ii) induction of metabolic compensatory adaptations, with significant shift to glycolysis; iii) induction of different antioxidant enzyme activities; iv) significant cell growth inhibition and v) no significant apoptosis. This work will permit better characterization the mitochondrial damage induced by hyperoxia. In particular, the data showed a large increase in the succinate cytochrome c reductase activity, which could be a fundamental pathogenic mechanism at the basis of oxygen toxicity.

Metabolic Abnormalities in Patients with Mitochondrial Myopathy Evaluated by In Vivo $^{31}$P Magnetic Resonance Spectroscopy (인($^{31}$P) 자기공명분광법을 사용하여 사립체 근질병환자와 정상인과의 대사물질 비교조사)

  • Bo-Young Choe
    • Investigative Magnetic Resonance Imaging
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    • v.2 no.1
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    • pp.89-95
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    • 1998
  • Purpose : To investigate the phosphorus metabolic abnormalities in skeletal muscle of patients with mitochondrial myopathy using in vivo $^{31}P$ magnetic resonance spectroscopy(MRS). Materials and Methods : Patients with mitochondrial myopathy(N=10) and normal control subjects (N=10) participated. All in vivo $^{31}P$ MRS examinations were performed on 1.5T whole-body MRI/MRS system by using an image selected in vivo spectroscopy (ISIS) pulse sequence that provided a $4{\times}4{\times}4{\;}cm^{3}$ volume of interest (VOI) in the right thigh muscle tissue. Peak areas for each phophorus methabolite were measured using a Marquart algorithm. Results : The specific features in patients with mitochondrial myopathy were a significant increase of Pi/PCr ratio (p=0.003) and a significant decrease of ATP/PCr ratio (p=0.004) as compared with normal controls. In particular, the ${\beta}-ATP/PCr$ ratio between controls and patients with mitochondrial myopathy was predominantly altered. Conclusions : In vivo $^{31}P$ MRS may be a useful modality in the clinical evaluation of patients with mitochondrial myopathy based on ATP/PCr and Pi/PCr ratios in skeletal muscle tissue and provides a valuable information in further understanding disorders of muscle metabolism.

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Cell Biological Study on Factors Affecting Brain Formation at Early Chick Embryo (1) The Effect of Serotonin (초기 계배의 뇌형성에 미치는 몇가지 요인에 관한 세포 생물학적 연구 (1) Serotonin의 영향)

  • 최임순;주상옥;주충노;오억수;신길상
    • The Korean Journal of Zoology
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    • v.32 no.1
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    • pp.55-73
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    • 1989
  • The effect of tryptophan or serotonin on the early stage of chick brain development has been morphologically investigated using an electron microscope. The electron micrographs of neural plate cells of 1-day chick embryo treated with tryptophan or serotonin showed irregularity, evagination and disruption of nuclear membrane and nuclear chromatin condenstation, nucleolar margination and segregation. Hypertrophy of stalks, vesicles and vaculoes were seen and dilated and disrupted rough endoplasmic reticulum and underdeveloped neurotubules were also observed. In mesenchyme cells of tryptophan or serotonin administered 18 hr embryo, irregular nuclear membrane, swollen mitochondria, dilated rough endoplasmic reticulum and very large yolk granules were observed. Furthermore, DNA, RNA and protein contents of the embryos treated with typtophan or serotonin were considerably lower than those of control group. The amount of tubulin of the experimental groups was also greatly lower than that of control, suggesting that the impairment of microtubule formation occurred. Tryptophan or serotonin administration might depress the biosynthesis, of nucleic acid and protein including some enzymes tested. It seems that the serotonin formed from exogeneous tryptophan might inhibit the degradation of yolk granule by feedback regulation mechanism so as to impair microtububle and microvilli formation followed by a malformation of chick embryos.

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SYNERGISTIC APOPTOTIC EFFECT OF TAXOL ON ORAL SQUAMOUS CELL CARCINOMA BY CYCLOSPORIN A (구강 편평세포암종에서 Taxol과 Cyclosporin A의 세포사멸 상승 작용 효과)

  • Suh, Min-Jung;Han, Se-Jin;Lee, Jae-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.5
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    • pp.394-404
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    • 2007
  • Oral squamous cell carcinoma is the most prevalent oral cancer, which is characterized by its high metastasis and recurrent rates and poor prognosis. Taxol is an anticancer agent which is microbial products extracted from jew tree. It combines with the tubulin and induces apoptosis by inhibiting mitosis of cell with microtubule stabilization. Recently, it was reported to be effective in various solid tumors, but only very slight effect has been seen in oral squamous cell carcinomas due to its cell-specific potencies. Cyclosporin A is used as immune suppressant and is being applied in anticancer therapy as its mechanism of induction of change of apoptotic process in various cells have been known. In this study, oral squamous cell carcinoma HN22 cell line was used for in vitro experiment and as for the experimental group taxol and cyclosporin A were applied alone and to observe the synergistic effect of apoptosis, Taxol and cyclosporin A were coadministered with different concentration of taxol for comparison. The results were obtained as follow: 1. There was no difference in Bcl-2, Bax, caspase 3, 8, 9 mRNA expression when cyclosprin A or taxol was applied alone to HN 22 cell line. 2. Caspase 3, 9 mRNA expression was prominently increased when cyclosprin A and taxol were applied together to cancer cell. 3. No significant difference was observed when cyclosporin A and taxol($1{\mu}g/ml$ and $3{\mu}g/ml$) were applied together to cancer cell line. 4. No significant difference was seen in Bcl-2, Bax, and caspase 8 mRNA expression in all the groups of in vitro experiments. 5. When cyclosporin A was applied alone in vivo study on the nude mice, histopathologi cal findings was similar to those of the control group. Oral squamous cell carcinoma induced by inoculation of HN 22 cell line was not reduced after treatment of cyclosporin A. 6. When taxol was applied alone, the islands of squamous cell carcinoma still remained, which meant insignificant healing effect. There was a lesser volume increase compared with the cyclosporin A alone. 7. When taxol and cyclosporin A were applied together, the connective tissue and calcification were seen in the histopathologic findings. Oral squamous cell carcinoma was decreased and cancer cell was disappeared. In observing the tumor mass change with time, there was a gradual decreased size and healing features. As the results of the in vitro experiment, it could conclud that only when the two agents are applied together, mitochondria-mediated apoptosis occurred by considerable increase of caspase 3, 9 mRNA expression, irrespectable of the concentration of taxol. In vivo experiment, there was a discrete synergistic effect when the two agents were applied together. But single use of cyclosporin A was not effective in this study. Based on the results of this experiment, if further clinical studies are done, taxol and cyclosporin A could be effectively used in treatment of oral squamous cell carcinomas.

Stereoisomer-specific ginsenoside 20(S)-Rg3 reverses replicative senescence of human diploid fibroblasts via Akt-mTOR-Sirtuin signaling

  • Yang, Kyeong-Eun;Jang, Hyun-Jin;Hwang, In-Hu;Hong, Eun Mi;Lee, Min-Goo;Lee, Soon;Jang, Ik-Soon;Choi, Jong-Soon
    • Journal of Ginseng Research
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    • v.44 no.2
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    • pp.341-349
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    • 2020
  • Background: The replicative senescence of human dermal fibroblasts (HDFs) is accompanied by growth arrest. In our previous study, the treatment of senescent HDFs with Rg3(S) lowered the intrinsic reactive oxygen species (ROS) levels and reversed cellular senescence by inducing peroxiredoxin-3, an antioxidant enzyme. However, the signaling pathways involved in Rg3(S)-induced senescence reversal in HDFs and the relatedness of the stereoisomer Rg3(R) in corresponding signaling pathways are not known yet. Methods: We performed senescence-associated β-galactosidase and cell cycle assays in Rg3(S)-treated senescent HDFs. The levels of ROS, adenosine triphosphate (ATP), and cyclic adenosine monophosphate (cAMP) as well as the mitochondrial DNA copy number, nicotinamide adenine dinucleotide (NAD)+/1,4-dihydronicotinamide adenine dinucleotide (NADH) ratio, and NAD-dependent sirtuins expression were measured and compared among young, old, and Rg3(S)-pretreated old HDFs. Major signaling pathways of phosphatidylinositol 3-kinase/Akt, 5' adenosine monophosphate-activated protein kinase (AMPK), and sirtuin 1/3, including cell cycle regulatory proteins, were examined by immunoblot analysis. Results: Ginsenoside Rg3(S) reversed the replicative senescence of HDFs by restoring the ATP level and NAD+/NADH ratio in downregulated senescent HDFs. Rg3(S) recovered directly the cellular levels of ROS and the NAD+/NADH ratio in young HDFs inactivated by rotenone. Rg3(S) mainly downregulated phosphatidylinositol 3-kinase/Akt through the inhibition of mTOR by cell cycle regulators like p53/p21 in senescent HDFs, whereas Rg3(R) did not alter the corresponding signaling pathways. Rg3(S)-activated sirtuin 3/PGC1α to stimulate mitochondrial biogenesis. Conclusion: Cellular molecular analysis suggests that Rg3(S) specifically reverses the replicative senescence of HDFs by modulating Akt-mTOR-sirtuin signaling to promote the biogenesis of mitochondria.

Studies on the Cellular Metabolism in Microorganisms as Influenced by Gamma-irradiation.(II) - On the Respiration Rate and Dehydrogenase Actibity in Yeast Cells Irradiated by $\gamma$-ray. (미생물의 세포생리에 미치는 전이방사선의 영향에 관한 연구 (제 2보) - 효모균의 산소호흡기및 탈수소효소능에 대한 $\gamma$-ray 의 영향)

  • 김종협
    • Korean Journal of Microbiology
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    • v.5 no.2
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    • pp.69-78
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    • 1967
  • Kim, Jong Hyup, (Div. of Biology, Atomic Energy Research Institute.) Studies on the Cellular Metabolism in Microorganisms as influenced by Gamma-irradiation(II). On respiration rate and dehydrogenase activity of yeast cells irradiated by gamma ray from cobalt-60. 1. Oxygen uptake rate of the gamma irraiated yeast cells had been measured with Warburg's manometer, and the $O_{2}$-uptake was compared with those of normal cells. The rate of endogetious respiration increases in its $O_2$-uptake at 150, 000 rentgen dose, and at higher rentoen doses it was decreased. Exogenous respiration begin to decrease in its O_2$-uptake at 5, 000r. doses of irradiation, further decrease with increasing of doses unproportionally. 2. It appears that plasma-membrane and nuclear membrane of yeast cells have changed and denatured by gamma-irradiation, as exogenous respiration of glucose had been decreased at a dose of 200, 000r's irradiation. 3. The activity of glucose, alcoholic, lactic, succinic and glutamic deliydrogenase (G.D.H., A.D.H., L.D.H., S.D.11., and GL.D.H.) in the gamma irradaited cells had been assayed by T.T.C.(Triphenyl tetrazolium chloride) method and spectrophotometry, the obtained results were compared with those of normal cells. 4. At a dose of and 10, 000 rentgens' irradiation of gamma ray, the activty of each debydrogenase (G.D.H., A.D.H., L.D.H., ) shows a sharp and highest peak in optical absorbalicy, but each abtivity of S.D.H and Gl.D.H shows its' maximum peak at a dose of 30, 000r. 5. The curve of each dehydrogenase activity was found to be rhythmical according to dose-rate of gamma irradiation. 6. Comparing with activity of debydrogenase each other, the maximum peak in optical absorbency can be arranged according to order as follows; glucose > alcoholoic > lactic > glutamic > succinic, this order is identical to the order of breakdown utility in respiration of normal yeast cells. 7. The activity of dehydrogenase experimented exhibit a resistance against gamma irradiation at lethal dose of cells, and the activity of dehydrogenase are found to be much resistant than those of respiratory system. We may assume that the membrane substrate of mitochondria or cytoplasm had been destructed by gamma-irradiation much more than that of dehydronase system.

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Induction of Spontaneous Neutrophil Apoptosis by 4-O-Methyl-Ascochlorin, A Prenyl Phenol Compound (프레닐 페놀계 항생제인 4-O-methyl-ascochlorin에 의한 호중구 세포사멸의 유도)

  • Son Dong-Aoon;Lee Sun-Young;Lee Min-Jung;Park Joo-In;Hong Young-Seob;Lee Yong-Hwan;Chang Young-Chae;Kwak Jong-Young
    • Journal of Life Science
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    • v.16 no.1
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    • pp.30-36
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    • 2006
  • Neutrophils are short-lived leukocytes that play a vital role in immune responses to bacteria, yeast, and fungi. This study was performed to investigate the effect of 4-O-methyl-ascochlorin (MAC), an anti-tumor, antibiotic, and anti-fungal prenyl-phenol compound on the spontaneous apoptosis of human neutrophils. MAC time- and dose-dependently accelerated the spontaneous apoptosis of human neutrophils. The effect of MAC on neutrophil apoptosis was blocked by pre-treatment of the neutrophils with specific inhibitors of pancaspase (zVAD-fmk), caspase-8 (zIETD-fmk), or caspase-3 (zDEVD-fmk). The cleavage of procaspase-8 and procaspase-3 was increased by MAC. Mitochondrial permeability, which was measured by the retention of $DiOC_6(3)$, was dose-de-pendently increased by MAC but the change of mitochondrial permeability was not blocked by pretreatment of neutrophils with zIETD-fmk. These results suggest that MAC induces neutrophil apoptosis by caspase-8-dependent but mitochondria-independent manner.

Association of a Methanol Extract of Rheum undulatum L. Mediated Cell Death in AGS Cells with an Intrinsic Apoptotic Pathway

  • Hong, Noo Ri;Park, Hyun Soo;Ahn, Tae Seok;Jung, Myeong Ho;Kim, Byung Joo
    • Journal of Pharmacopuncture
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    • v.18 no.2
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    • pp.26-32
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    • 2015
  • Objectives: Rheum undulatum L. has traditionally been used for the treatment of many diseases in Asia. However, its anti-proliferative activity in cancer has still not been studied. In the present study, we investigated the anti-cancer effects of methanol extract of Rheum undulatum L. (MERL) on human adenocarcinoma gastric cell lines (AGS). Methods: To investigate the anti-cancer effect of MERL on AGS cells, we treated the AGS cells with varying concentrations of MERL and performed 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays. Cell cycle analyses, measurements of the mitochondrial membrane potential (MMP), caspase activity assays and Western blots were conducted to determine whether AGS cell death occurred by apoptosis. Results: Treatment with MERL significantly inhibited growth of AGS cells in a concentration dependent manner. MERL treatment in AGS cells leaded to increased accumulation of apoptotic sub G1 phase cells in a concentration dependent manner. In control cultures, 5.38% of the cells were in the sub G1 phase. In MERL treated cells, however, this percentage was significantly increased (9.95% at $70{\mu}g/mL$, 15.94% at $140{\mu}g/mL$, 26.56% at $210{\mu}g/mL$ and 38.08% at $280{\mu}g/mL$). MERL treatment induced the decreased expression of pro-caspase-8 and -9 in a concentration dependent manner, whereas the expression of the active form of caspase-3 was increased. A subsequent Western blot analysis revealed increased cleaved levels of poly (ADP-ribose) polymerase (PARP) protein. Also, treatment with MERL increased the activities of caspase-3 and -9 compared with the control. MERL treatment increased the levels of the pro-apoptotic truncated Bid (tBid) and Bcl2 Antagonist X (Bax) proteins and decreased the levels of the anti-apoptotic B-cell lymphoma 2 (Bcl-2) protein, whose is the stabilization of mitochondria. However, inhibitions of p38, extracellular signal regulated kinases (ERKs) and C-Jun N-terminal kinases (JNK) by MERL treatment did not affect cell death. Conclusion: These results suggest that MERL mediated cell death is associated with an intrinsic apoptotic pathway in AGS cells.

Ultrastructure of Gametes in the Three-spine stickleback, Gasterosteus aculeatus aculeatus (큰가시고기 배우자의 미세구조)

  • Deung, Young-Kun;Kim, Dong-Heui;Reu, Dong-Suck
    • Applied Microscopy
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    • v.29 no.2
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    • pp.177-187
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    • 1999
  • Ultrastructure of gametes in the three-spine stickleback, Gasterosteus aculeatus aculeatus was observed, utilizing light, scanning and transmission electron microscopes. The egg of three-spine stickleback is spherical and demersal type. The eggs are highly adhesived to each other but not to substrates. There are many oil droplets in vitelline membrane. The outer surface of egg envelope is arranged by mushroom-like structures and pore canals. The egg have a micropyle, sperm entry site, in the area of the animal pole. The egg envelope consists of three layers, an outer layer with high electron density, a middle layer consisting two layers and an inner layer consisting of 16 to 20 layers. In the fertilized egg envelope, the molecular weights of these components ranged from 14 kDa to 205 kDa. The molecular weights of nam protein bands are 19.4 kDa, 36.7 KDa, 39.4 kDa, 42.9 kDa, 46.1 kDa and 53.0 kDa. The head of spermatozoa is spherical shape and the acrosome is absent. The mitochondria in midpiece are arranged from one to three layers and separated from the axoneme by the cytoplasmic canal. The tail has two lateral fins and the axoneme is of the 9+2 structure.

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