• Title/Summary/Keyword: cellular DNA content

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Flooding Stress-Induced Glycine-Rich RNA-Binding Protein from Nicotiana tabacum

  • Lee, Mi-Ok;Kim, Keun Pill;Kim, Byung-gee;Hahn, Ji-Sook;Hong, Choo Bong
    • Molecules and Cells
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    • v.27 no.1
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    • pp.47-54
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    • 2009
  • A cDNA clone for a transcript preferentially expressed during an early phase of flooding was isolated from Nicotiana tabacum. Nucleotide sequencing of the cDNA clone identified an open reading frame that has high homology to the previously reported glycine-rich RNA-binding proteins. The open reading frame consists of 157 amino acids with an N-terminal RNA-recognition motif and a C-terminal glycine-rich domain, and thus the cDNA clone was designated as Nicotiana tabaccum glycine-rich RNA-binding protein-1 (NtGRP1). Expression of NtGRP1 was upregulated under flooding stress and also increased, but at much lower levels, under conditions of cold, drought, heat, high salt content, and abscisic acid treatment. RNA homopolymer-binding assay showed that NtGRP1 binds to all the RNA homopolymers tested with a higher affinity to poly r(G) and poly r(A) than to poly r(U) and poly r(C). Nucleic acid-binding assays showed that NtGRP1 binds to ssDNA, dsDNA, and mRNA. NtGRP1 suppressed expression of the fire luciferase gene in vitro, and the suppression of luciferase gene expression could be rescued by addition of oligonucleotides. Collectively, the data suggest NtGRP1 as a negative modulator of gene expression by binding to DNA or RNA in bulk that could be advantageous for plants in a stress condition like flooding.

The Change of Secretory Activity of the Alveolar Type ll Cell During Acute Alveolar Injury Induced by N-Nitroso-N-Methylurethane

  • Lee, Young-Man;Bang, In-Sook;Lee, Suck-Kang
    • The Korean Journal of Physiology
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    • v.28 no.1
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    • pp.71-77
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    • 1994
  • In the animal model of acute respiratory distress syndrome (ARDS) induced by N-nitroso-N-methylurethane (NNNMU) the secretory activity of alveolar type H cells during acute alveolar injury was investigated by determining phospholipid and pulmonary surfactant associated proteins in crude surfactant. The mechanism of the secretory change was studied by determination of DNA and RNA levels in the lung tissue. After induction of acute alveolar injury with NNNMU, pulmonary hemorrhage, atelectasis and gross hypertrophy were observed. Seven days after NNNMU treatment the level of total DNA in lung homogenate was increased markedly indicating that a hypertrophy was induced by cellular proliferation. Although the total DNA level increased, the RNA/DNA ratio was gradually decreased after NNNMU treatment. Seven days after NNNMU treatment the RNA/DNA ratio returned to the normal control level. During the acute alveolar injury, phospholipid and surfactant associated proteins were reduced significantly as compared with the control, implying that the secretory activity of alveolar type II cells was altered during acute alveolar injury induced by NNNMU. The protein content in crude surfactant during peak injury(7 days after NNNMU) was decreased significantly but phospholipid/protein ratios were identical in both control and NNNMU treatment groups. SDS-PAGE of proteins in crude pulmonary surfactant showed a decrease in major surfactant associated protein(M.W. 38,000) during acute alveolar injury. The present study may suggest that while alveolar type H cells proliferate markedly, transcription of alveolar type ll cell gene was inhibited by an unknown mechanism such as DNA methylation induced by NNNMU. Such an inhibition of transcriptional activity is thought to be associated with the decreased secretory activity of alveolar type ll cells, which may lead to pulmonary atelectasis and edema during the acute alveolar injury.

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The Effect of Injin and Injinsaryungsangagambang on Liver Cell Viability, Lever Cell Cycle Progression and DNA Damage-induced Apoptosis (인진(茵蔯)과 인진사령산가감방(茵蔯四岺散加減方)이 간세포활성(肝細胞活性), 세포주기(細胞週期) 및 DNA damage-induced apoptosis에 미치는 영향(影響))

  • Kang, Woo-Sung;Lee, Jang-Hoon;Woo, Hong-Jung
    • The Journal of Korean Medicine
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    • v.20 no.1 s.37
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    • pp.91-105
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    • 1999
  • The effects of Yinjin and Yinjinsaryongsangagambang on a DNA damaging agent, etoposide-induced apoptosis, cell viability, cell cycle progression, and mRNA expression of apoptosis-related genes of human hepatocyte cell line HepG2 were investigated using tryphan blue exclusion assay, MTT assay, flow cytometry, immunocytometric analysis of PCNA, and quantitative RT-PCR analysis. MTT assay showed that Yinjin and Yinjinsaryongsangagambang increases cellular viability of HepG2 cells in a dosage-dependent manner. Stimulation of cell cycle progression by Yinjin or Yinjinsaryongsangagambang was detected by flow cytometric analysis of the DNA content and immunocytometric analysis of PCNA expression. A significant reduction of a DNA-damaging agent, etoposide-induced apoptosis were found in both Yinjin and Yinjinsaryongsangagambang-treated cells in dosage-dependent manner. In overall, 3-fold reduction of apoptosis was recognized in $10.0\;{\mu}g/ml$ of Yinjin or Yinjinsaryongsangagambang-treated cells compared to untreated cells. Although the difference is not significant, Yinjinsaryongsangagambang showed slightly higher effect on the inhibition of apoptosis than Yinjin. From flow cytometric analysis of apoptosis, while 39.9% of untreated cells showed etoposide-induced apoptotic cell death, only 19.6% or 17.4% of Yinjin or Yinjinsaryongsangagambang-treated cells were fond at apoptotic sub G1 phase, respectively. Interestingly, strong induction of Gadd45-mRNA was observed from Yinjin or Yinjinsaryongsangagambang-treated cells. However, no changes in expression levels of p53 and Waf1 were detected, demonstrating that induction of Gadd45 mRNA expression by Yinjin or Yinjinsaryongsangagambang occurs by p53-independent mechanism. Marked mRNA inductions of two apoptosis-inhibiting genes, Bcl-2 and Bcl- XL, were found in both Yinjin or Yinjinsaryongsangagambang-treated HepG2 cells while no changes was detected in expression levels of an apoptosis-promoting gene, Bax.

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Antioxidant Activity of Orange Flesh and Peel Extracted with Various Solvents

  • Park, Jae-Hee;Lee, Minhee;Park, Eunju
    • Preventive Nutrition and Food Science
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    • v.19 no.4
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    • pp.291-298
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    • 2014
  • The aim of this study was to investigate the antioxidant activity of orange (Citrus auranthium) flesh (OF) and peel (OP) extracted with acetone, ethanol, and methanol. Antioxidant potential was examined by measuring total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (RSA), total radical-trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), and cellular antioxidant activity (CAA). The comet assay was used to determine the protective effects of OF and OP against $H_2O_2$-induced DNA damage. TPC was highest in the acetone extracts of OF and OP. DPPH RSA was also higher in the acetone extracts than in the ethanol extracts. The DPPH RSA was highest in the acetone extracts of OF. The TRAP and ORAC values of the all extracts increased in a dose-dependent manner. In the TRAP assay, the acetone extracts of OF and OP had the lowest $IC_{50}$ values. In the CAA assay, the methanol and acetone extracts of OP had the lowest $IC_{50}$ values. All of the samples protected against $H_2O_2$-induced DNA damage in human leukocytes, as measured by the comet assay, but the acetone extracts of OP had the strongest effect. These results suggest that acetone is the best solvent for the extraction of antioxidant compounds from OF and OP. Furthermore, the high antioxidant activity of OP, which is a by-product of orange processing, suggests that it can be used in nutraceutical and functional foods.

Chemical Characteristics and Antioxidant Effects of Sea Mustard Undaria pinnatifida from the Gultongmeori Area, Taejongdae, Busan (부산 태종대 굴통머리 미역(Undaria pinnatifida)의 화학적 특성 및 항산화 효과)

  • Young Do Shin;Jung Woo Lee;Myungwon Choi;Sun Young Lim
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.2
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    • pp.196-203
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    • 2023
  • We investigated the nutritional characteristics and antioxidant effects of sea mustard Undaria pinnatifida fractions from Gultongmeori in Taejongdae, Youngdo, Busan. Based on dry weight, the moisture, crude protein, crude lipid, crude ash, and crude fiber contents were 34.98%, 11.55%, 0.43%, 17.82%, and 3.45%, respectively. To evaluate the antioxidant effect, we used radical scavenging (DPPH and ABTS), reactive oxygen species (ROS) production measurement, and DNA oxidation assays. Total flavonoid and phenol contents were higher in the n-hexane fraction than in other fractions. The n-hexane fraction was more effective at scavenging free radicals than other fractions as assessed using DPPH and ABTS assays (P<0.05). The ROS production assay showed that all sea mustard fractions decreased H2O2 induced cellular ROS production compared to that seen in the control (P<0.05); however, the n-hexane fraction reduced cellular ROS production to a greater extent than the other fractions. Furthermore, the n-hexane fraction from Gultongmeori significantly inhibited genomic DNA oxidation. These results indicate that the antioxidant effect of sea mustard is associated with its high flavonoid and phenol content. This study proposes that processed food products supplemented with sea mustard can be developed as functional foods to promote health in the local population.

Effects of Heterologous Expression of Thioredoxin Reductase on the Level of Reactive Oxygen Species in COS-7 Cells

  • Kang, Hyun-Jung;Hong, Sung-Min;Kim, Byung-Chul;Park, Eun-Hee;Ahn, Kisup;Lim, Chang-Jin
    • Molecules and Cells
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    • v.22 no.1
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    • pp.113-118
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    • 2006
  • Thioredoxin reductase (TrxR), a component of the redox control system involving thioredoxin (Trx), is implicated in defense against oxidative stress, control of cell growth and proliferation, and regulation of apoptosis. In the present study a stable transfectant was made by introducing the vector pcDNA3.0 harboring the fission yeast TrxR gene into COS-7 African green monkey kidney fibroblast cells. The exogenous TrxR gene led to an increase in TrxR activity of up to 3.2-fold but did not affect glutathione (GSH) content, or glutaredoxin and caspase-3 activities. Levels of reactive oxygen species (ROS), but not those of nitric oxide (NO), were reduced. Conversely, 1-chloro-2,4-dinitrobezene (CDNB), an irreversible inhibitor of mammalian TrxR, enhanced ROS levels in the COS-7 cells. After treatment with hydrogen peroxide, the level of intracellular ROS was lower in the transfectants than in the vector control cells. These results confirm that TrxR is a crucial determinant of the level of cellular ROS during oxidative stress as well as in the normal state.

Epigenetics: A key paradigm in reproductive health

  • Bunkar, Neha;Pathak, Neelam;Lohiya, Nirmal Kumar;Mishra, Pradyumna Kumar
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.2
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    • pp.59-81
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    • 2016
  • It is well established that there is a heritable element of susceptibility to chronic human ailments, yet there is compelling evidence that some components of such heritability are transmitted through non-genetic factors. Due to the complexity of reproductive processes, identifying the inheritance patterns of these factors is not easy. But little doubt exists that besides the genomic backbone, a range of epigenetic cues affect our genetic programme. The inter-generational transmission of epigenetic marks is believed to operate via four principal means that dramatically differ in their information content: DNA methylation, histone modifications, microRNAs and nucleosome positioning. These epigenetic signatures influence the cellular machinery through positive and negative feedback mechanisms either alone or interactively. Understanding how these mechanisms work to activate or deactivate parts of our genetic programme not only on a day-to-day basis but also over generations is an important area of reproductive health research.

Random Sequence Analysis of the Genomic DNA of Methanopyrus kandleri and Molecular Cloning of the Gene Encoding a Homologue of the Catalytic Subunit of Carbon Monoxide Dehydrogenase

  • Shin, Hyun-Seock;Ryu, Jae-Ryeon;Han, Ye-Sun;Choi, Yong-Jin;Yu, Yeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.404-413
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    • 1999
  • Methanopyrus kandleri is a hyperthermophilic methanogen that represents one of the most heat-resistant organisms: the maximum growth temperature of M. kandleri is $110^{\circ}C$. A random sequence analysis of the genomic DNA of M. kandleri has been performed to obtain genomic information. More than 200 unique sequence tags were obtained and compared with the sequences in the GenBank and PIR databases. About 30% of the analyzed tags showed strong sequence similarity to previously identified genes involved in various cellular processes such as biosynthesis, transport, methanogenesis, or metabolism. When statistics relating to the frequency of codons were examined, the sequenced open reading frames showed highly biased codon usage and a high content of charged amino acids. Among the identified genes, a homologue of the catalytic subunit of carbon monoxide dehydrogenase (CODH) that reduces $CO_2$ to CO was cloned and sequenced in order to examine its detailed gene structure. The cloned gene includes consensus promoters. The amino acid sequence of the cloned gene shows a strong homology with the CODH genes from methanogenic Archaea, especially in the presumed binding sites for Fe-S centers.

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Characterization of a New Acidophilic Acetobacter sp. Strain HA Isolated from Korean Traditional Fermented Vinegar

  • CHUN, HONG-SUNG;SUNG-JUN KIM
    • Journal of Microbiology and Biotechnology
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    • v.3 no.2
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    • pp.108-114
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    • 1993
  • A new strain of acidophilic, acetogenic bacterium, Acetobacter sp. strain HA was isolated by selective enrichment from the traditionally fermented rice wine vinegar in Korea. It was a gram-negative, non-motile short rod and oxidized acetate and lactate. The optimal temperature and pH for growth were $28^{\circ}C$ and 4.0, respectively. The strain HA differed from other Acetobacter species by growing well on methanol, xylitol, inositol, dulcitol, D-xylose, L-arabinose, and D-mannose as sole sources of carbon and energy. The isolated strain HA did not produce $\gamma$-pyrones from glucose and did not produce ketone bodies from glycerol. The quinone system used in this study was an ubiquinone-9 isoprene unit. The guanine-plus-cytosine content of the DNA was 50.7 mol%, and the major cellular fatty acids were $C_{18:1} and C_{16:0}$.

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Cytogenetic Analysis of Induced Hybrid between Common Carp (Cyprinus carpio) and Crucian Carp (Carassius auratus) (잉어(Cyprinus carpio)와 붕어(Carassius auratus)간 잡종의 세포유전학적 분석)

  • 남윤권;오승용;조재윤;김동수
    • Journal of Aquaculture
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    • v.11 no.1
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    • pp.77-81
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    • 1998
  • Cytogenetic analyses were carried out with induced hybrid between common carp (Cyprinus carpio) female and crucian carp (Carassius auratus) male. The erythrocytic measurement revealed that cellular and nuclear size of induced hybrids were intermediate between those of paremtal species. The modal chromosome numbers of common carp, crucian carp and its hybid were same as 2n=100. The DNA content of induced hybrids determined based on flow cytometry was 3.7pg/cell which corresponding to intermediate value between the carp (3.6ph/cell) and crucian carp (3.8pg/cell)

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