• Title/Summary/Keyword: Mitochondrial complex I

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Characterization of Lupinus Iuteus Chloroplgsl Gene Coding for Components of a Chloroplastic NADH Dehydrogenase

  • Oczkowski, Marian;Augustyniak, Halina
    • Journal of Plant Biotechnology
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    • v.2 no.2
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    • pp.73-78
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    • 2000
  • The plastid genomes of several plants contain ndh genes homologues of genes encoding subunits of the mitochondrial complex I. We sequenced the part of lupin ndhB, ndhD and ndhF genes in order to compare the structure of these genes with those of Nicotiana tabaum, Arabidopsis thaliana, Zea mays and Oryza sativa with the idea to detect the presence of stretches with identical aminoacid composition. We were only able to find one or two stretches of this kind of about 16 aminoacid- long in the analyzed fragments of the ndh genes. The total number of such stretches was different in particular gene products: for ndhc 1, ndhB 9, ndhD 3 and ndhF 6. We have also examined the transcription pattern of ndhC, ndhK and ndhJ genes during lupin development. We show that the greatest amount of ndhC, ndhK and ndhJ transcripts are observed in 7- to 14 day- old lupin seedlings. We also studied the level of transcription of those genes in plants growing at low temperature. All the data confirmed that the abundance of transcription of ndhC, ndhK, and ndhJ genes increased under chill conditions. It has to be noted that the level of transcription of the ndhC gene was higher than the other genes probably due to higher stability of this transcript.

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Naringin Protects against Rotenone-induced Apoptosis in Human Neuroblastoma SH-SY5Y Cells

  • Kim, Hak-Jae;Song, Jeong-Yoon;Park, Hae-Jeong;Park, Hyun-Kyung;Yun, Dong-Hwan;Chung, Joo-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.4
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    • pp.281-285
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    • 2009
  • Rotenone, a mitochondrial complex I inhibitor, can induce the pathological features of Parkinson's disease (PD). In the present study, naringin, a grapefruit flavonoid, inhibited rotenone-induced cell death in human neuroblastoma SH-SY5Y cells. We assessed cell death and apoptosis by measuring mitogen-activated protein kinase (MAPKs) and caspase (CASPs) activities and by performing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Naringin also blocked rotenone-induced phosphorylation of Jun NH2-terminal protein kinase (JNK) and P38, and prevented changes in B-cell CLL/lymphoma 2 (BCL2) and BCL2-associated X protein (BAX) expression levels. In addition, naringin reduced the enzyme activity of caspase 3 and cleavages of caspase 9, poly (ADP-ribose) polymerase (PARP), and caspase 3. These results suggest that naringin has a neuroprotective effect on rotenone-induced cell death in human neuroblastoma SH-SY5Y cells.

NADPH Oxidase and Mitochondrial ROS are Involved in the $TNF-{\alpha}$-induced Vascular Cell Adhesion Molecule-1 and Monocyte Adhesion in Cultured Endothelial Cells

  • Yu, Jae-Hyeon;Kim, Cuk-Seong;Yoo, Dae-Goon;Song, Yun-Jeong;Joo, Hee-Kyoung;Kang, Gun;Jon, Ji-Yoon;Park, Jin-Bong;Jeon, Byeong-Hwa
    • The Korean Journal of Physiology and Pharmacology
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    • v.10 no.4
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    • pp.217-222
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    • 2006
  • Atherosclerosis is considered as a chronic inflammatory process. However, the nature of the oxidant signaling that regulates monocyte adhesion and its underlying mechanism is poorly understood. We investigated the role of reactive oxygen species on the vascular cell adhesion molecule-1 (VCAM-1) and monocyte adhesion in the cultured endothelial cells. $TNF-{\alpha}$ at a range of $1{\sim}30\;ng/ml$ induced VCAM-1 expression dose-dependently. BCECF-AM-labeled U937 cells firmly adhered on the surface of endothelial cells when the endothelial cells were incubated with $TNF-{\alpha}$ (15 ng/ml). Ten $\;{\mu}mol/L$ of SB203580, an inhibitor of p38 MAPK, significantly reduced $TNF-{\alpha}-induced$ VCAM-1 expression, compared to the JNK inhibitor ($40\;{\mu}mol/L$ of SP60015) or ERK inhibitor ($40\;{\mu}mol/L$ of U0126). Also, SB203580 significantly inhibited $TNF-{\alpha}-induced$ monocyte adhesion in HUVEC. Superoxide production was minimal in the basal condition, however, treatment of $TNF-{\alpha}$ induced superoxide production in the dihydroethidineloaded endothelial cells. Diphenyleneiodonium (DPI, $10\;{\mu}mol/L$), an inhibitor of NADPH oxidase, and rotenone $(1\;{\mu}mol/L)$, an inhibitor of mitochondrial complex I inhibited $TNF-{\alpha}-induced$ superoxide production, VCAM-1 expression and monocyte adhesion in the endothelial cells. Taken together, our data suggest that NADPH oxidase and mitochondrial ROS were involved in $TNF-{\alpha}-induced$ VCAM-1 and monocyte adhesion in the endothelial cells.

Comprehensive investigations of key mitochondrial metabolic changes in senescent human fibroblasts

  • Ghneim, Hazem K.;Alfhili, Mohammad A.;Alharbi, Sami O.;Alhusayni, Shady M.;Abudawood, Manal;Aljaser, Feda S.;Al-Sheikh, Yazeed A.
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.4
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    • pp.263-275
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    • 2022
  • There is a paucity of detailed data related to the effect of senescence on the mitochondrial antioxidant capacity and redox state of senescent human cells. Activities of TCA cycle enzymes, respiratory chain complexes, hydrogen peroxide (H2O2), superoxide anions (SA), lipid peroxides (LPO), protein carbonyl content (PCC), thioredoxin reductase 2 (TrxR2), superoxide dismutase 2 (SOD2), glutathione peroxidase 1 (GPx1), glutathione reductase (GR), reduced glutathione (GSH), and oxidized glutathione (GSSG), along with levels of nicotinamide cofactors and ATP content were measured in young and senescent human foreskin fibroblasts. Primary and senescent cultures were biochemically identified by monitoring the augmented cellular activities of key glycolytic enzymes including phosphofructokinase, lactate dehydrogenase, and glycogen phosphorylase, and accumulation of H2O2, SA, LPO, PCC, and GSSG. Citrate synthase, aconitase, α-ketoglutarate dehydrogenase, succinate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, and complex I-III, II-III, and IV activities were significantly diminished in P25 and P35 cells compared to P5 cells. This was accompanied by significant accumulation of mitochondrial H2O2, SA, LPO, and PCC, along with increased transcriptional and enzymatic activities of TrxR2, SOD2, GPx1, and GR. Notably, the GSH/GSSG ratio was significantly reduced whereas NAD+/NADH and NADP+/NADPH ratios were significantly elevated. Metabolic exhaustion was also evident in senescent cells underscored by the severely diminished ATP/ADP ratio. Profound oxidative stress may contribute, at least in part, to senescence pointing at a potential protective role of antioxidants in aging-associated disease.

Neurotoxin-Induced Pathway Perturbation in Human Neuroblastoma SH-EP Cells

  • Do, Jin Hwan
    • Molecules and Cells
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    • v.37 no.9
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    • pp.672-684
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    • 2014
  • The exact causes of cell death in Parkinson's disease (PD) remain unknown despite extensive studies on PD.The identification of signaling and metabolic pathways involved in PD might provide insight into the molecular mechanisms underlying PD. The neurotoxin 1-methyl-4-phenylpyridinium ($MPP^+$) induces cellular changes characteristic of PD, and $MPP^+$-based models have been extensively used for PD studies. In this study, pathways that were significantly perturbed in $MPP^+$-treated human neuroblastoma SH-EP cells were identified from genome-wide gene expression data for five time points (1.5, 3, 9, 12, and 24 h) after treatment. The mitogen-activated protein kinase (MAPK) signaling pathway and endoplasmic reticulum (ER) protein processing pathway showed significant perturbation at all time points. Perturbation of each of these pathways resulted in the common outcome of upregulation of DNA-damage-inducible transcript 3 (DDIT3). Genes involved in ER protein processing pathway included ubiquitin ligase complex genes and ER-associated degradation (ERAD)-related genes. Additionally, overexpression of DDIT3 might induce oxidative stress via glutathione depletion as a result of overexpression of CHAC1. This study suggests that upregulation of DDIT3 caused by perturbation of the MAPK signaling pathway and ER protein processing pathway might play a key role in $MPP^+$-induced neuronal cell death. Moreover, the toxicity signal of $MPP^+$ resulting from mitochondrial dysfunction through inhibition of complex I of the electron transport chain might feed back to the mitochondria via ER stress. This positive feedback could contribute to amplification of the death signal induced by $MPP^+$.

Fine Structure on the Spermiogenesis of Octopus minor on the Western Coast of Korea I (한국 서해안 서해낙지 (Octopus minor)의 정자 완성에 관한 미세구조 I)

  • Chang, Nam-Sub;Kim, Sang-Won;Han, Jong-Min
    • Applied Microscopy
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    • v.31 no.3
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    • pp.223-233
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    • 2001
  • The spermiogenesis of a Korean octopus, Octopus minor, inhabiting western of Korea Sea was observed by electron microscopy . The obtained results are as follows: The spermiogenesis of Octopus miner proceeds through four stages; early- , mid- , and late-spermatid, and mature sperm. An early spermatid is a spherical cell looking light due to the low electron density. The acrosome formed from Golgi complex of the upper nucleus looks dark due to the high electron density. The extra-nuclear rod (enr) stemming from proximal centriole is transformed from round shape into oval shape, elongating to the upper nucleus. In our observation, the axoneme was being formed from distal centriole, and the manchette composed of a number of microtubules is also found around nuclear membrane. In a mid-spermatid, chromatins in the nucleus contract shaping fine threads, and the manchette is also observed around nuclear membrane. Especially, the spherical acrosome is transformed into long oval one which is tinged with a number of horizontal stripes and has the middle electron density. In a late-spermatid, chromatins in the nucleus contract thick and short. Furthermore, the mitochondrial sleeve, in which the axoneme is surrounded with mitochondria, is observed at middle piece. The axoneme has a typical structure of 9+2 and around it, 9 coarse fibers are observed. Also in the acrosome cavity of mature sperm, horizontal striation is found. However, regularly spaced processes are peculiarly observed in there. A sperm is about 390 fm long, whose head is bent a little like a banana while the acrosome region is helical. In the middle piece of sperm, $11\sim12$ mitochondria are surrounding coarse fibers that reach the main piece of tail, while nothing but 9+2 structured axoneme is found in the end piece.

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Intraspecific diversity and phylogeography of bony lip barb, Osteochilus vittatus, in Sundaland, as revealed by mitochondrial cytochrome oxidase I (mtCOI)

  • Imron Imron;Fajar Anggraeni;Wahyu Pamungkas;Huria Marnis;Yogi Himawan;Dessy Nurul Astuti;Flandrianto Sih Palimirmo;Otong Zenal Arifin;Jojo Subagja;Daniel Frikli Mokodongan;Rahmat Hidayat
    • Fisheries and Aquatic Sciences
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    • v.27 no.3
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    • pp.145-158
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    • 2024
  • Life history characteristics, habitat landscape, and historical events are believed to have shaped the patterns of genetic variation in many taxa. The bony lip barb, Osteohilus vittatus, represent a potamodromous fish that complete all life cycle in freshwater and is widely distributed in Southeast Asia. It usually lives in small rivers and other freshwater habitats, and movement between habitats for either food or reproduction has been typical. These life history characteristics may promote gene flow, leading to less structured populations. However, many freshwater habitats are fragmented, which restricts gene flow. We investigate how this interplay has shaped patterns of genetic variation and phylogeographic structure within this species in the Sundaland, a biodiversity hotspot with a complex geological history, using mitochondrial cytochrome oxidase I (mtCOI) as a genetic marker. Forty-six mtCOI sequences of 506 bp long were collected from ten localities, eight geographically isolated and two connected. The sequences were used for population genetic and phylogeographic analyses. Our results showed a low genetic diversity within populations but high between populations. There was a deep phylogeographic structure among geographically isolated populations but a lack of such structure in the connected habitats. Among geographically isolated populations, sequence divergence was revealed, ranging from 1.8% between Java and Sumatra populations to 12.2% between Malaysia and Vietnam. An indication of structuring was also observed among localities that are geographically closer but without connectivity. We conclude that despite high dispersal capacity, the joint effects of historical events, long-term geographic isolation associated with sea level oscillation during the Pleistocene, and restricted gene flow related to lack of habitat connectivity have shaped the phylogeographic structure within the O. vittatus over the Sundaland.

Genetic Diversity and Pathogenicity of Cylindrocarpon destructans Isolates Obtained from Korean Panax ginseng

  • Song, Jeong Young;Seo, Mun Won;Kim, Sun Ick;Nam, Myeong Hyeon;Lim, Hyoun Sub;Kim, Hong Gi
    • Mycobiology
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    • v.42 no.2
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    • pp.174-180
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    • 2014
  • We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.

Inherited metabolic diseases in the urine organic acid analysis of complex febrile seizure patients (복합 열성경련 환자의 소변 유기산 분석에서 나타난 유전대사질환)

  • Cheong, Hee Jeong;Kim, Hye Rim;Lee, Seong Soo;Bae, Eun Joo;Park, Won Il;Lee, Hong Jin;Choi, Hui Chul
    • Clinical and Experimental Pediatrics
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    • v.52 no.2
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    • pp.199-204
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    • 2009
  • Purpose : Seizure associated with fever may indicate the presence of underlying inherited metabolic diseases. The present study was performed to investigate the presence of underlying metabolic diseases in patients with complex febrile seizures, using analyses of urine organic acids. Method : We retrospectively analyzed and compared the results of urine organic acid analysis with routine laboratory findings in 278 patients referred for complex febrile seizure. Results : Of 278 patients, 132 had no abnormal laboratory findings, and 146 patients had at least one of the following abnormal laboratory findings: acidosis (n=58), hyperammonemia (n=55), hypoglycemia (n=21), ketosis (n=12). Twenty-six (19.7 %) of the 132 patients with no abnormal findings and 104 (71.2%) of the 146 patients with statistically significant abnormalities showed abnormalities on the organic acid analysis (P<0.05). Mitochondrial respiratory chain disorders (n=23) were the most common diseases found in the normal routine laboratory group, followed by PDH deficiency (n=2) and ketolytic defect (n=1). In the abnormal routine laboratory group, mitochondrial respiratory chain disorder (n=29) was the most common disease, followed by ketolytic defects (n=27), PDH deficiency (n=9), glutaric aciduria type II (n=9), 3-methylglutaconic aciduria type III (n=6), biotinidase deficiency (n=5), propionic acidemia (n=4), methylmalonic acidemia (n=2), 3-hydroxyisobutyric aciduria (n=2), orotic aciduria (n=2), fatty acid oxidation disorders (n=2), 2-methylbranched chain acyl CoA dehydrogenase deficiency (n=2), 3-methylglutaconic aciduria type I (n=1), maple syrup urine disease (n=1), isovaleric acidemia (n=1), HMG-CoA lyase deficiency (n=1), L-2-hydroxyglutaric aciduria (n=1), and pyruvate carboxylase deficiency (n=1). Conclusion : These findings suggest that urine organic acid analysis should be performed in all patients with complex febrile seizure and other risk factors for early detection of inherited metabolic diseases.

Inhibition of Proliferation and Neurogenesis of Mouse Subventricular Zone Neural Stem Cells by a Mitochondrial Inhibitor Rotenone (미토콘드리아 억제제 rotenone에 의한 쥐의 뇌실 하 영역 신경 줄기 세포의 증식과 신경 세포로의 분화 억제)

  • Park, Ki-Youb;Kim, Man Su
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1397-1405
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    • 2018
  • Mitochondria have multiple functions in cells: providing chemical energy, storing cellular $Ca^{2+}$, generating reactive oxygen species, and regulating apoptosis. Through these functions, mitochondria are also involved in the maintenance, proliferation, and differentiation of stem/progenitor cells. In the brain, the subventricular zone (SVZ) is one of the neurogenic regions that contains neural stem cells (NSCs) throughout a lifetime. However, reports on the role of mitochondria in SVZ NSCs are scarce. Here, we show that rotenone, a complex I inhibitor of mitochondria, inhibits the proliferation and differentiation of SVZ NSCs in different ways. In proliferating NSCs, rotenone decreases mitosis as measured through phosphorylated histone H3 detection; moreover, apoptosis is not induced by rotenone at 50 nM. In differentiating NSCs, rotenone blocks neurogenesis and oligodendrogenesis while glial fibrillary acidic protein-positive astrocytes are not affected. Interestingly, in this study there were more cells in the differentiating NSCs treated with rotenone for 4-6 days than in the vehicle control group which was a different effect from the reduced number of cells in the proliferating NSCs. We examined both apoptosis and mitosis and found that rotenone decreased apoptosis as detected by staining cleaved caspase-3 but did not affect mitosis. Our results suggest that functional mitochondria are necessary in both the proliferation and differentiation of SVZ NSCs. Furthermore, mitochondria might be involved in the mitosis and apoptosis that occur during those processes.