• Title/Summary/Keyword: CytA

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Effects of Potassium Ion and Caffeine on Contraction and Cytosolic Free $Ca^{2+}$ Levels in Vascular Smooth Muscle (혈관평할근 세포에서의 칼륨이온과 카페인의 영향: 수축과 세포내 칼슘이온 농도에 대하여)

  • Ahn, H.Y.;Karaki, H.
    • The Korean Journal of Pharmacology
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    • v.24 no.2
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    • pp.197-201
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    • 1988
  • Effects of high concentration of KC1 and caffeine on cytosolic $Ca^{2+}$ level $([Ca^{2+}]_{cyt})$, measured simultaneously with muscle tension using a fluorescent intracellular $Ca^{2+}$ indicator fura 2, were examined in isolated smooth muscle of rat aorta. High $K^+$ (72.7 mM) solution induced sustained increase in both $([Ca^{2+}]_{cyt})$ and tension. In contrast to this, caffeine (20 mM) induced a rapid increase in $([Ca^{2+}]_{cyt})$ followed by a decrease to a level which was higher than the resting level. However, muscle tension showed only a transient increase followed by a decrease below the resting level. In a $Ca^{2+}-free$ solution, high $K^+-induced$ neither $([Ca^{2+}]_{cyt})$ nor tension, whereas caffeine induced a transient increase in both $([Ca^{2+}]_{cyt})$ and muscle tension. These results suggest that high $K^+-induced$ contraction in vascular smooth muscle of rat aorta is due to $Ca^{2+}$ influx whereas caffeine-induced contraction is due to $Ca^{2+}$ release from cellular store. Further, caffeine seems to have an additional effect to decrease the sensitivity of the contractile elements to $Ca^{2+}$.

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Induction of the Intrinsic Apoptotic Pathway by 3-Deazaadenosine Is Mediated by BAX Activation in HL-60 Cells

  • Lee, Sun-Young;Ko, Kyoung-Won;Kang, Won-Kyung;Choe, Yun-Jeong;Kim, Yoon-Hyoung;Kim, In-Kyung;Kim, Jin;Kim, Ho-Shik
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.6
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    • pp.407-412
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    • 2010
  • 3-Deazaadenosine (DZA), a potent inhibitor of S-adenosylhomocysteine hydrolase, was previously proposed to induce intrinsic apoptosis in human leukemic cells. In the present study, we analyzed the mechanism underlying the DZA-induced intrinsic apoptotic pathway. DZA activated typical caspase-dependent apoptosis in HL-60 cells, as demonstrated by an accumulation of hypo-diploidic cells, the processing of multiple procaspases and an inhibitory effect of z-VAD-Fmk on this cell death. During DZA-induced apoptosis, cytochrome c (cyt c) was released into the cytosol. This was neither prevented by z-VAD-Fmk and nor was it associated with the dissipation of mitochondrial membrane potential (${\Delta}{\Psi}_m$). Prior to the release of cyt c, BAX was translocated from the cytosol to mitochondria and underwent oligomerization. Finally, the overexpression of BCL-XL protected HL-60 cells from apoptosis by blocking both the cyt c release and BAX oligomerization. Collectively, these findings suggest that DZA may activate intrinsic apoptosis by stimulating BAX activation and thereby the release of cyt c.

Mechanism of Fatty Acid Synthase in Drug Tolerance Related to Epithelial-mesenchymal Transition of Breast Cancer

  • Li, Jun-Qin;Xue, Hui;Zhou, Lan;Dong, Li-Hua;Wei, Da-Peng;Li, Hua
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7617-7623
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    • 2014
  • Objective: The mechanism of action of fatty acid synthase (FASN) in drug tolerance of breast cancer cells with epithelial-mesenchymal transition (EMT) features was investigated. Methods: The breast cancer cell line MCF-7-MEK5 with stably occurring EMT and tumour necrosis factor-${\alpha}$ (TNF-${\alpha}$) tolerance was used as the experimental model, whereas MCF-7 acted as the control. Tumour cells were implanted into nude mice for in vivo analysis, and cerulenin was used as a FASN inhibitor. RT-PCR, real-time quantitative PCR and Western blot were employed to detect the expression of FASN, TNFR-1, TNFR-2, Wnt-1, ${\beta}$-catenin and cytC at the RNA and protein levels. Results: Compared with MCF-7, TNFR-1 expression in MCF-7-MEK5 was slightly changed, TNFR-2 was decreased, and FASN, Wnt-1, ${\beta}$-catenin and cytC were increased. The expression of Wnt-1 and ${\beta}$-catenin in MCF-7-MEK5 decreased after cerulenin treatment, whereas cytC expression increased. Conclusions: The important function of FASN in the drug tolerance of breast cancer may be due to the following mechanisms: FASN downregulated TNFR-2 expression through lipid rafts to make the cells less sensitive to TNF-${\alpha}$, and simultaneously activated the Wnt-$1/{\beta}$-catenin signalling pathway. Thus, cytC expression increased, which provided cells with anti-apoptotic capacity and induced drug tolerance.

The Genetic Approach on Analyzing the Habitat Characteristics of Fairy Pitta Pitta Nympha Inhabiting Jeju Island, the Korean Peninsula and Taiwan (제주도, 한반도 및 대만 내 팔색조의 생태적 서식특성 분석을 위한 유전적 접근)

  • Kim, Eun-Mi;Jeon, Yeon-Seon;Kim, Se-Jae;Kang, Chang-Wan;Won, Hyun-Kyu;Jeong, Gil-Sang
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.17 no.1
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    • pp.81-90
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    • 2014
  • A Fairy Pitta is a bird known to breed only in mainland China, Taiwan, Japan and Korea and is listed as Vulnerable in the IUCN Red List. We carried out a DNA analysis to contribute to conserve the genetic diversity of Fairy Pitta. 32 samples were collected at Jeju Island, the Korean Peninsula and Taiwan from 2004 to 2013 and DNA was extracted from them and several sequences were amplified-it through PCR. And then we performed the population genetic analysis. We found there was a transversion between nucleotide sequences at CO1 gene, while there was no changes at Cyt-b gene. And we confirmed the polymorphism from two genes was caused from genetic drift not from selection. Through this analysis, the group within the Peninsula was found bigger than other two groups based on the analysis of CO1 gene, and the group from Taiwan was found bigger than other two groups through the analysis of Cyt-b gene. The population genetic structure of mitochondria gene of three group was showing CO1 gene had 5 haplotypes and Cyt-b gene had 6 haplotypes. Haplotype 2 in CO1 gene was found in three group and many individuals of samples had this haplotype. Like CO1 gene, haplotype 2 in Cyt-b gene was found in three group and was included in plenty of individuals. Other haplotypes were not overlaped and broke off among the three groups. To prevent from the extinction of Fairy Pitta and to obtain the genetic diversity, we need to compare with other regional group such as Japan, China and perform additional research in the non-breeding area.

Investigation of the electrode reaction of cytochrome c and pyrroliquinoline quinone at self-assembled monolayers of amino acid

  • Kim Imsook;Kwak Juhyoun
    • Journal of the Korean Electrochemical Society
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    • v.2 no.1
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    • pp.27-30
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    • 1999
  • Self-Assembled monolayers of carboxyl-terminated alkanethiols, which is negatively charged in pH 7.0, were usually used to facilitate the electron transfer between the positively charged protein and the electrode. In case of L-cysteine, as it has both positive and negative group, it can be a candidate for a new modifier to facilitate positively charged protein or negatively charged protein. Our investigation of L-cysteine shows that the electron transfer occurs successfully to both cytochrome c (cyt. c) and pyrroloquinoline quinone (PQQ). By using 1-ethyl-3-(3-dime-thlyaminopropyl) carbodiimide (EDC), we made a covalent bond between cyt. c and monolayer. Then PQQ was electrostatically adsorbed to the same monolayer. Cyclic voltammograms show that both molecules do not interfere each other and electron transfer is appreciable.

Diversity of Bacillus thuringiensis Strains Isolated from Citrus Orchards in Spain and Evaluation of Their Insecticidal Activity Against Ceratitis capitata

  • J.C., Vidal-Quist;Castanera, P.;Gonzalez-Cabrera, J.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.749-759
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    • 2009
  • A survey of Bacillus thuringiensis (Berliner) strains isolated from Spanish citrus orchards has been performed, and the strains were tested for insecticidal activity against the Mediterranean fruit fly Ceratitis capitata (Wiedemann), a key citrus pest in Spain. From a total of 150 environmental samples, 376 isolates were selected, recording a total B. thuringiensis index of 0.52. The collection was characterized by means of phase-contrast microscopy, SDS-PAGE, and PCR analysis with primer pairs detecting toxin genes cry1, cry2, cry3, cry4, cry5, cry7, cry8, cry9, cry10, cry11, cry12, cry14, cry17, cry19, cry21, cry27, cry39, cry44, cyt1, and cyt2. Diverse crystal inclusion morphologies were identified: bipyramidal (45%), round (40%), adhered to the spore (7%), small (5%), and irregular (3%). SDS-PAGE of spore-crystal preparations revealed 39 different electrophoresis patterns. All primer pairs used in PCR tests gave positive amplifications in strains of our collection, except for primers for detection of cry3, cry19, cry39, or cry44 genes. Strains containing cry1, cry2, cry4, and cry27 genes were the most abundant (48.7%, 46%, 11.2%, and 8.2% of the strains, respectively). Ten different genetic profiles were found, although a total of 109 strains did not amplify with the set of primers used. Screening for toxicity against C. capitata adults was performed using both spore-crystal and soluble fractions. Mortality levels were less than 30%. We have developed a large and diverse B. thuringiensis strain collection with huge potential to control several agricultural pests; however, further research is needed to find out Bt strains active against C. capitata.

Comparison of the Genetic Relationships and Osteological Aspects in Six Branchiostegid Fish Species (Perciformes)

  • Ryu, Jung-Hwa;Kim, Jin-Koo;Park, Jung-Youn
    • Animal cells and systems
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    • v.13 no.3
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    • pp.323-329
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    • 2009
  • We analyzed partial sequences of cytochrome b (cyt-b), a mitochondrial DNA (mtDNA) gene, to determine the genetic relationships between six horsehead fish species: Branchiostegus japonicus, Branchiostegus albus, Branchiostegus auratus, Branchiostegus argentatus, Branchiostegus wardi, and an unidentified Branchiostegus species. The specimens were collected in Korea, China, Japan, and Vietnam. We compared their molecular phylogenetic relationships inferred from mtDNA cyt-b sequences with an osteological analysis. The unidentified species, B. sp., was similar to B. albus in terms of the lack of triangular silver-white dot at the posterior region of eyes (vs. large one present in B. japonicus), but was also similar to B. japonicus in terms of the presence of a straight-shaped first hemal spine (vs. a curve-shaped hemal spine in B. albus). Analysis of the mtDNA cyt-b sequences indicated that the smallest estimated sequence divergence was between the B. japonicus and B. sp. (0.70-0.94%), whereas the largest difference was between B. auratus and B. argentatus (23.06-23.36%). Both the maximum parsimony and maximum likelihood trees showed that the B. sp. was closely clustered with B. japonicus, and that B. auratus was most distant from the other species. When comparing the osteological characters, UPGMA tree showed that the B. japonicus and B. sp. were the most closely clustered species, and B. auratus was the most distantly clustered fish relative to the other species. The shape of the nasal, otolith and first hemal spine was informative for distinguishing B. auratus from the other species. These osteological differences were consistent with the differences in mtDNA.

Phylogenetic Characterization of White Hanwoo Using the Mitochondrial Cytochrome b Gene (mtDNA cytochrome b 분석을 통한 백한우의 계통유전학적 특성 분석)

  • Kim, Jae-Hwan;Cho, ChangYeon;Kim, SeungChang;Kim, Sung Woo;Choi, Seong-Bok;Lee, Seong-Su
    • Journal of Life Science
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    • v.25 no.9
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    • pp.970-975
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    • 2015
  • The goals of this study were to identify sequence variations in the mitochondrial cytochrome b (mtDNA cyt b) gene in White Hanwoo (Wh) and the genetic relationship between the Wh and other breeds. When whole sequences of the mtDNA cyt b gene in 14 Wh cattle were determined, a silent mutation and two haplotypes were detected in the Wh cattle. The major haplotype, H1, was found in 13 of 14 individuals in the Wh cattle. Haplotype diversity and nucleotide diversity were 0.143 and 0.00013, respectively. Compared to previous reports, these levels of genetic diversity are lower than other Korean and Chinese breeds. To identify the genetic relationship among Korean, Chinese, Japanese, and European cattle breeds, the neighbor-joining (NJ) tree was constructed based on Dxy genetic distances. Two distinct groups were identified and classified as A and B. Wh was found in the A group, which consisted of Bos taurus breeds. From calculating the Dxy genetic distances, Wh was found to be genetically more closely related to two breeds, Heugu (0.00018) and Yanbian (0.00021), than to other breeds. In conclusion, Wh is genetically related to Chikso, Heugu, and Yanbian breeds based on maternal inheritance. The results of this study will be useful for efficient management and sustainable utilization of Wh.

Genetic Relationships of Sandfish (Arctoscopus japonicas) from Five Different Areas of Korea and Japan Based on Mitochondrial DNA and Microsatellite Analyses (Mitochondrial DNA와 microsatellite marker 분석을 통한 한국과 일본에 서식하는 5 지역의 도루묵(Arctoscopus japonicas)에 대한 유전학적 유연관계 분석)

  • Kim, Eun-Mi;Kang, Hyun-Sook;Kang, Jung-Ha;Kim, Dong-Gyun;An, Cheul Min;Lee, Hae Won;Park, Jung Youn
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1204-1213
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    • 2015
  • A comprehensive analysis of the population structure of the sandfish (Arctoscopus japonicas), the most abundant fishery resource in the East Sea of Korea, has not been carried out, despite its importance in Korea. The present study examined the genetic diversity and differences between five populations (two Japanese and three Korean populations) of A. japonicas captured in the East Sea using both the 401 bp sequence of mitochondrial DNA (mtDNA, cytochrome b) and five microsatellite DNA (msDNA) markers. The results of the analysis using the Cyt b sequence revealed 27 haplotypes. Based on msDNA variations, the estimated expected heterozygosity (HE) in each population ranged from 0.68 (Gampo, Korea) to 0.7765 (Erimo, Japan). Pairwise FST and AMOVA tests using both the Cyt b sequence and msDNA data pointed to significant differences between the Korean and Japanese populations (mtDNA; FST=0.2648, p<0.05, msDNA; FST=0.0814, p<0.05). These results were similar to the results of UPGMA, PCA, and structure analysis. In these analyses, the five populations were assigned to two groups (Korean populations and Japanese populations). These results shed light on the genetic diversity and relationships of A. japonicas and contribute to research on the evaluation, conservation, and utilization of Korean A. japonicas as genetic resources.

Molecular Phylogeny of Two Species of Hexagrammidae (Greenlings) Inferred from Mitochondrial Cytochrome b Gene (쥐노래미과 어류 2종의 미토콘드리아 Cytochrome b 유전자의 분자계통)

  • Jung, Sang-Oun;Lee, Young-Mi;Hur, Jun-Wook;Im, Soo-Yeon;Lee, Jae-Seong;Park, In-Seok
    • Korean Journal of Environmental Biology
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    • v.24 no.2 s.62
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    • pp.112-118
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    • 2006
  • We report mitochondrial cytochrome b (cyt b) genes from greenling Hexagrammos otakii (Jordan et Starks) and spotty belly greenling, H. agrammus (Temminck et Schlegel) within Hexagrammidae, two species of aquaculture importance. Of 489 bp of the cytochrome b gene, a little variation occurred between species (96% similarity). The pairwise distance (0.0342) between greenling and spotty belly greenling in term of the Neighbor-joining method indicated that two species was close in molecular phylogenetic consideration. These findings are applicable to aquaculture, fisheries genetics and molecular phylogenetics in the genus Hexagrammos.