• Title/Summary/Keyword: Mitochondrial DNA

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Cloning, Expression and Hormonal Regulation of Steroidogenic Acute Regulatory Protein Gene in Buffalo Ovary

  • Malhotra, Nupur;Singh, Dheer;Sharma, M.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.2
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    • pp.184-193
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    • 2007
  • In mammalian ovary, steroidogenic acute regulatory (StAR) protein mediates the true rate-limiting step of transport of cholesterol from outer to inner mitochondrial membrane. Appropriate expression of StAR gene represents an indispensable component of steroidogenesis and its regulation has been found to be species specific. However, limited information is available regarding StAR gene expression during estrous cycle in buffalo ovary. In the present study, expression, localization and hormonal regulation of StAR mRNA were analyzed by semi-quantitative RT-PCR in buffalo ovary and partial cDNA was cloned. Total RNA was isolated from whole follicles of different sizes, granulosa cells from different size follicles and postovulatory structures like corpus luteum and Corpus albicans. Semi-quantitative RT-PCR analyses showed StAR mRNA expression in the postovulatory structure, corpus luteum. No StAR mRNA was detected in total RNA isolated from whole follicles of different size including the preovulatory follicle (>9 mm in diameter). However, granulosa cells isolated from preovulatory follicles showed the moderate expression of StAR mRNA. To assess the hormonal regulation of StAR mRNA, primary culture of buffalo granulosa cells were treated with FSH (100 ng/ml) alone or along with IGF-I (100 ng/ml) for 12 to 18 h. The abundance of StAR mRNA increased in cells treated with FSH alone or FSH with IGF-I. However, effect of FSH with IGF-I on mRNA expression was found highly significant (p<0.01). In conclusion, differential expression of StAR messages was observed during estrous cycle in buffalo ovary. Also, there was a synergistic action of IGF-I on FSH stimulation of StAR gene.

Occurrence Pattern of an Unidentified Moth Captured by Sex Pheromone Trap of the Oriental Fruit Moth, Grapholita molesta, and Its Discriminating Molecular Markers (복숭아순나방(Grapholita molesta) 성페로몬 트랩에 포획된 미동정 나방의 발생패턴과 판별 분자지표)

  • Huh, Hye-Jung;Son, Ye-Rim;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.47 no.3
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    • pp.303-308
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    • 2008
  • An unidentified moth was captured in sex pheromone traps of the oriental fruit moth, Grapholita molesta, especially at spring season in apple orchards and their vicinity. Though the captured males were similar in appearance to G. molesta males, they were easily distinguished by a matted difference in body size. Their occurrence pattern was also similar to that of overwintering G. molesta population from April to May, at which more males were captured in the pheromone traps installed in the vicinity of apple orchards than within apple orchards. After May, they were no longer captured in the pheromone traps. To investigate any larval damage due to this unidentified moth, molecular markers needed to be developed. Four PCR-RFLP markers originated from cytochrome b region of mitochondrial DNA could distinguish this unidentified moth from G. molesta.

Predominant Genotypes and Alleles of Two Functional Polymorphisms in the Manganese Superoxide Dismutase Gene are Not Associated with Thai Cervical or Breast Cancer

  • Attatippaholkun, Watcharee;Wikainapakul, Kornwipa
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3955-3961
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    • 2013
  • Background: Defects of manganese superoxide dismutase (MnSOD) have long been implicated in generation of oxidative stress and risk susceptibility to various cancers. Two functional polymorphisms within the MnSOD gene, including the Val-9Ala of the mitochondrial targeting sequence (MTS) and the Ile58Thr of the exon-3, have been proposed to reduce its enzyme activity and antioxidant potential. Materials and Methods: A high-throughput multiplex SNaPshot$^{(R)}$ system was developed herein for simultaneous analyses of Val-9Ala and Ile58Thr in a single reaction. Genomic DNA extracted from each whole blood sample of 248 patients including 107 with cervical cancer and 141 with breast cancer and from 136 healthy women as controls was analyzed by the multiplex SNaPshot$^{(R)}$ system. Results: The Val/Val, Val/Ala genotypes and the Val allele of the MTS were predominant in patients with cervical or breast cancer as well as healthy women in Thailand. The Ile/Ile genotype and the Ile allele of the exon-3 were found in all of them whereas none of the Ile/Thr, the Thr/Thr genotypes and the Thr allele was detected. Genotypic association of both Val-9Ala and Ile58Thr polymorphisms with cervical cancer and breast cancer of these patients comparing to healthy women was not statistically significant (p<0.05). Conclusions: The Val/Val, Val/Ala genotypes and the Val allele of the MTS were found predominantly but the Ile/Ile genotype and the Ile allele of the exon-3 were detected in patients with cervical cancer, breast cancer and healthy women in Thailand. These two functional polymorphisms (Val-9Ala and Ile58Thr) in MnSOD gene did not associate with susceptibility risk of these cancer patients in Thailand.

Molecular Identification of Diphyllobothrium latum from a Pediatric Case in Taiwan

  • An, Yu-Chin;Sung, Chia-Cheng;Wang, Chih-Chien;Lin, Hsin-Chung;Chen, Kuang-Yao;Ku, Fu-Man;Chen, Ruei-Min;Chen, Mei-Li;Huang, Kuo-Yang
    • Parasites, Hosts and Diseases
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    • v.55 no.4
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    • pp.425-428
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    • 2017
  • Human diphyllobothriasis is a parasitic disease caused by ingestion of larvae (plerocercoids) in raw or undercooked fish and commonly found in temperate areas. Rare cases were reported in tropical or subtropical areas especially in children. The first documented case of pediatric diphyllobothriasis in Taiwan had been reported 11 years ago. Here, we report another 8-year-old girl case who presented with a live noodle-like worm hanging down from her anus, with no other detectable symptoms. We pulled the worm out and found the strobila being 260 cm in length. Examination of gravid proglottids showed that they were wider than their lengths, containing an ovoid cirrus sac in the anterior side and the rosette-shaped uterus. Eggs extracted from the uterus were ovoid and operculated. Diphyllobothrium latum was confirmed by molecular analysis of the mitochondrial DNA cytochrome c oxidase subunit 1 (cox1) gene. The girl was treated with a single oral dose of praziquantel, and no eggs or proglottids were observed from her stool in the subsequent 3 months. The reemergence of human diphyllobothriasis in non-endemic countries is probably due to prevalent habit of eating imported raw fish from endemic areas. This pediatric case raised our concern that human diphyllobothriasis is likely underestimated because of unremarkable symptoms.

Occurrence of skipjack tuna (Katsuwonus pelamis) larvae in Korean waters (가다랑어 (Katsuwonus pelamis) 자어의 국내 출현)

  • Yoon, Sang Chul;Jeong, Yeon Kyu;Ji, Hwan Sung;Shin, Ari;Kim, Zang Geun;Choi, Kwang Ho
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.51 no.1
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    • pp.79-85
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    • 2015
  • Three specimens of larvae (5.2~7.8 mm in standard length (SL)), of the family Scombridae, were collected from the southeastern waters off Jeju Island in August, 2014. These specimens were identified to Katsuwonus pelamis have melanophores distributed on the 1st dorsal-fin spines and post ventral margin on caudal peduncle. An analysis of the cytochrome oxidase submit I (COI) sequences (652 base pairs) of mitochondrial DNA showed that our specimens must be K. pelamis, because their sequences were concordant with those of the adult K. pelamis (genetic distance= 0.000~0.002), followed by Auxis rochei (genetic distance= 0.061~0.063) and Euthynnus affinis (genetic distance= 0.077~0.079). During the larval stage, K. pelamis differed from Thunnus spp. species in melanophores distribution period of 1st dorsal-fin spines, lower jaws and ventral margin on caudal peduncle. In conclusion, occurrence of K. pelamis larvae means Korean waters are potential spawning ground of K. pelamis.

A Natural Product, Chios Gum Mastic, Induces the Death of HL-60 Cells via Apoptosis and Cell Cycle Arrest

  • Koo, Byung-Chan;Kim, Duck-Han;Kim, In-Ryoung;Kim, Gyoo-Cheon;Kwak, Hyun-Ho;Park, Bong-Soo
    • International Journal of Oral Biology
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    • v.36 no.1
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    • pp.13-21
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    • 2011
  • Chios gum mastic (CGM) is produced from Pistiacia lentiscus L var chia, which grows only on Chios Island in Greece. CGM is a kind of resin extracted from the stem and leaves, has been used for many centuries in many Mediterranean countries as a dietary supplement and folk medicine for stomach and duodenal ulcers. CGM is known to induce cell cycle arrest and apoptosis in some cancer cells. This study was undertaken to investigate the alteration of the cell cycle and induction of apoptosis following CGM treatment of HL-60 cells. The viability of the HL-60 cells was assessed using the MTT assay. Hoechst staining and DNA electrophoresis were employed to detect HL-60 cells undergoing apoptosis. Western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, MMP activity and proteasome activity analyses were also employed. CGM treatment of HL-60 cells was found to result in a dose- and time-dependent decrease in cell viability and apoptotic cell death. Tested HL-60 cells showed a variety of apoptotic manifestations and induced the downregulation of G1 cell cycle-related proteins. Taken collectively, our present findings demonstrate that CGM strongly induces G1 cell cycle arrest via the modulation of cell cycle-related proteins, and also apoptosis via proteasome, mitochondrial and caspase cascades in HL-60 cells. Hence, we provide evidence that a natural product, CGM could be considered as a novel therapeutic for human leukemia.

Protective Effects of Sosokmyoung-tang Against Parkinson's Model in Human Neuroblastoma SH-SY5Y Cells (사람 신경모세포종 SH-SY5Y 세포주의 파킨슨 모델에 대한 소속명탕(小續命湯)의 보호효과)

  • Woo, Chan;You, Ju-Yeon;Jang, Chul-Yong;Kim, Hyo-Rin;Shin, Yong-Jeen;Moon, A-Ji;Shin, Sun-Ho
    • The Journal of Internal Korean Medicine
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    • v.35 no.3
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    • pp.298-308
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    • 2014
  • Objectives: In this study we made an effort to investigate the protective effect of SSMT on the N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) -induced cytotoxicity of SH-SY5Y cells. Methods: The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MMT) assay. The fluorescence intensity was measured by using a dye and then with propidium iodide (PI) DNA flow cytometry analysis of the effects on the cell cycle of the SH-SY5Y cells and were used to measure the fluorescence of intracellular reactive oxygen species generation by MPTP. Results: Pretreatment of SSMT significantly suppressed MPTP-induced cytotoxicity, which was revealed as apoptosis characterized by the reduction of cell viability, the increase of ROS production, and the loss of mitochondrial membrane potential in SH-SY5Y cells. Conclusions: These findings suggest that SSMT exerts neuroprotective effects on human neuroblastoma SH-SY5Y cells by MPTP-induced dopaminergic neurodegeneration.

Protective Effects of Hwangryunhaedog-tang on Hypoxia-induced Apoptosis in H9c2 Cardiomyoblast Cells (황연해독탕이 저산소증에 의한 배양심근세포고사에 미치는 영향)

  • Jeong Jae Eun;Yu Bong Seon;Park Jin Yeong;Jeon In Cheol;Park Sang Beom;Lee Dae Yong;Lee Min Goo;Lee In;Moon Byun Soon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1733-1739
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    • 2004
  • The water extract of Hwangryunhaedog-tang(HRHDT} has been traditionally used for treatment of ischemic heart and brain damage in oriental medicine. However, little is known about the mechanism by which the water extract of HRHDT rescues cells from these damages. This study was designed to investigate the protective mechanisms of HRHDT on hypoxia-induced cytotoxicity in H9c2 cardiomyoblast cells. Hypoxia, markedly decreased the viability of H9c2 cells, which was characterized with apparent apoptptic features such as chromatin condensation as well as fragmentation of genomic DNA and nuclei. However, HRHDT significantly reduced hypoxia-induced cell death and apoptotic characteristics. Also, HRHDT prevented the mitochondrial dysfunction including the disruption of mitochondria membrane permeability transition (MPT) and an increase in expression of anti-apoptotic Bcl-2 proteins in hypoxia-H9c2 cells. Taken together, this study suggests that the protective effects of the water extract of HRHDT against hypoxic damages may be mediated by the modulation of Bcl-2 and Bak expression.

Effects of Sibjeondaebo-Tang on Oxidative Stress of C6 Glial Cells (십전대보탕이 신경교세포의 산화적 손상에 미치는 영향)

  • Ryu Ji Yong;Yun Jong Min;Cho Kwang Ho;Moon Byung Soon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.4
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    • pp.1120-1128
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    • 2004
  • Zinc ion has both essential and toxic effects on mammalian cells. The results demonstrated that the ability of zinc to act as an inducer of apoptosis in C6 glial cells. Incubation with 0.2 mM ZnCl₂ caused cell death that was characterized as apoptosis by internucleosomal DNA fragmentation, formation of apoptotic bodies, nuclear fragmentation and breakdown of the mitochondrial membrane potential. ZnCl₂-induced apoptosis of C6 glial cells was prevented by the addition of Sibjeondaebo-Tang and antioxidants including reduced glutathione, N-acetyl-L-cysteine. We further confirmed that ZnCl₂ decreased the intracellular levels of GSH as well as generation of H₂O₂ in C6 glial cells. In 2D-electrophoresis, computer-assisted comparative analysis of the respective silver stained spot patterns revealed 3 groups with strongly decreased intensity by ZnCl₂. Whereas, 3 groups with increasing intensity were recovered by Sibjeondaebo-Tang. These results suggest that Sibjeondaebo-Tang may function as an antioxidant against free radicals and be applicable to the treatment of brain cells against oxidative stress.

Combination Effects of 7β-Hydroxycholesterol and Radiation in Human Lung Cancer Cells

  • KANG Kyoung Ah;LEE Kyoung Hwa;CHAE Sungwook;KIM Dae Yong;PARK Moon Taek;LEE Su Jae;LEE Yun Sil;HYUN Jin Won
    • Biomolecules & Therapeutics
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    • v.13 no.4
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    • pp.220-226
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    • 2005
  • The present study was performed to evaluate combination effect of 7$\beta$-hydroxycholesterol (7$\beta$-OHC) and $\gamma$-radiation in NCI-H460 human lung cancer cells. 7$\beta$-OHC in combination with $\gamma$-irradiation has an enhanced effect in decreasing clonogenic survival and increasing cellular DNA damage. Pretreatment of cells with 7$\beta$-OHC enhanced radiation-induced apoptosis. Apoptosis of the cells by combined treatment of 7$\beta$-OHC and $\gamma$-irradiation was associated with reactive oxygen species generation and loss of mitochondrial membrane potential, resulting in the activation of caspase 9 and caspase 3. The combined treatment also resulted in an increased G1 cell cycle distribution. These results indicate that 7$\beta$-OHC shows the additive effect of radiation sensitivity in human lung carcinoma cells in vitro.