• Title/Summary/Keyword: DNA-level

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Combined Genotoxic Effects of Aflatoxin B1, Ochratoxin A and Zearalenone in Rat Bone Marrow and Blood Leukocytes

  • Tigran, Harutyunyan;Anna, Karapetyan;Galina, Hovhannisyan;Rouben, Aroutiounian
    • Korean Journal of Environmental Biology
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    • v.31 no.3
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    • pp.189-191
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    • 2013
  • Mycotoxins such as aflatoxin B1 (AFB1), ochratoxin A (OTA) and zearalenone (ZEA) are widespread contaminants of food and feedstuffs. It is very likely, that humans and animals are always exposed to mixtures of mycotoxins rather than to individual compounds. Therefore, risk assessments should consider mixture toxicity data. In the present study the combination of AFB1, OTA and ZEA was tested for genotoxicity in rat bone marrow and blood leukocytes after 15, 30 and 60 days treatment. The level of DNA damage was determined by the comet assay. The tail intensity and Olive tail moment in leukocytes and bone marrow cells were significantly higher than in controls. At the same time, the level of DNA damage in bone marrow cells was higher than in leukocytes. The data suggests that prolonged exposure to mycotoxins combination through food consumption can induce DNA damage contributing to the harmful effects in vivo.

Exploring the Utility of Partial Cytochrome c Oxidase Subunit 1 for DNA Barcoding of Gobies

  • Jeon, Hyung-Bae;Choi, Seung-Ho;Suk, Ho Young
    • Animal Systematics, Evolution and Diversity
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    • v.28 no.4
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    • pp.269-278
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    • 2012
  • Gobiids are hyperdiverse compared with other teleost groups, with about 2,000 species occurring in marine, freshwater, and blackish habitats, and they show a remarkable variety of morphologies and ecology. Testing the effectiveness of DNA barcodes on species that have emerged as a result of radiation remains a major challenge in evolutionary biology. Here, we used the cytochrome c oxidase subunit 1 (COI) sequences from 144 species of gobies and related species to evaluate the performance of distance-based DNA barcoding and to conduct a phylogenetic analysis. The average intra-genus genetic distance was considerably higher than that obtained in previous studies. Additionally, the interspecific divergence at higher taxonomic levels was not significantly different from that at the intragenus level, suggesting that congeneric gobies possess substantial interspecific sequence divergence in their COI gene. However, levels of intragenus divergence varied greatly among genera, and we do not provide sufficient evidence for using COI for cryptic species delimitation. Significantly more nucleotide changes were observed at the third codon position than that at the first and the second codons, revealing that extensive variation in COI reflects synonymous changes and little protein level variation. Despite clear signatures in several genera, the COI sequences did resolve genealogical relationships in the phylogenetic analysis well. Our results support the validity of COI barcoding for gobiid species identification, but the utilization of more gene regions will assist to offer a more robust gobiid species phylogeny.

Isolation and Molecular Phylogeny of Three Muscle Actin Isoforms of an Endangered Freshwater Fish Species Hemibarbus mylodon (Cypriniformes; Cyprinidae)

  • Kim, Keun-Yong;Nam, Yoon-Kwon
    • Journal of Aquaculture
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    • v.22 no.1
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    • pp.83-91
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    • 2009
  • The Korean doty barbel Hemibarbus mylodon (Cypriniformes; Cyprinidae) is a critically endangered freshwater fish species mainly because of its natural habitat degradation. Three full-length complementary DNA (cDNA) clones representing different muscle actin isoforms were isolated and characterized. The three muscle actin isoforms were 1,294-1,601 bp long with the identical open reading frames of 1,134 bp with the deduced amino acid residues of 377. They showed 83.9-87.2% identities in the coding nucleotide level and 96.8-98.1% identities in the amino acid level. Phylogenetic analysis with the coding nucleotide sequences revealed that three muscle actin isoforms of H. mylodon formed strongly supported monophyletic groups with one of cypriniform skeletal $\alpha$-actin (acta1), cypriniform aortic $\alpha$-actins (acta2), and uncharacterized Danio rerio muscle actin isoform/Salmo trutta slow muscle actin (a novel muscle actin type). Our phylogenetic tree further suggested that cypriniform acta2 only showed the orthologous relationship to tetrapod acta2. Other multiple actin isoforms from diverse teleostean taxa were however clustered to no tetrapod orthologs, i.e., acta1, cardiac $\alpha$-actins (aetc1), acta2, and enteric $\gamma$-actin (actg2). This result strongly suggested that teleostean muscle actins have experienced different and complicated evolutionary history in comparison to mammalian counterparts.

Ubiquitin E3 ligases controlling p53 stability

  • Lee, Seong-Won;Seong, Min-Woo;Jeon, Young-Joo;Chung, Chin-Ha
    • Animal cells and systems
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    • v.16 no.3
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    • pp.173-182
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    • 2012
  • The p53 protein plays a pivotal role in tumor suppression. The cellular level of p53 is normally kept low by proteasome-mediated degradation, allowing cell cycle progression and cell proliferation. Under stress conditions, such as DNA damage, p53 is stabilized and activated through various post-translational modifications of itself as well as of its regulatory proteins for induction of the downstream genes responsible for cell cycle arrest, DNA repair, and apoptosis. Therefore, the level of p53 should be tightly regulated for normal cell growth and for prevention of the accumulation of mutations in DNA under stress conditions, which otherwise would lead to tumorigenesis. Since the discovery of Mdm2, a critical ubiquitin E3 ligase that destabilizes p53 in mammalian cells, nearly 20 different E3 ligases have been identified and shown to function in the control of stability, nuclear export, translocation to chromatin or nuclear foci, and oligomerization of p53. So far, a large number of excellent reviews have been published on the control of p53 function in various aspects. Therefore, this review will focus only on mammalian ubiquitin E3 ligases that mediate proteasome-dependent degradation of p53.

Platform of Hot Pepper Stress Genomics: Indentification of Stress Inducible Genes in Hot Pepper (Capsicum annuum L.) Using cDNA Microarray Analysis

  • Chung, Eun-Jo;Lee, Sanghyeob;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.81.1-81
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    • 2003
  • Although plants have evolved to possess various defense mechanisms from local biotic and abiotic stressors, most of yield loss is caused by theses stressors. Recent studies have revealed that several different stress responsive reactions are inter-networking. Therefore, the identification and dissection of stress responsive genes is an essential and first step towards understanding of the global defense mechanism in response to various stressors. For this purpose, we applied cDNA microarray analysis, because it has powerful ability to monitor the global gene expression in a specific situation. To date, more than 10,000 non-redundant genes were identified from seven different cDNA libraries and deposited in our EST database (http://plant.pdrs.re.kr/ks200201/pepper.html). For this study, we have built 5K cDNA microarray containing 4,685 unigene clones from three different cDNA libraries. Monitoring of gene expression profiles of hot pepper interactions with biotic stress, abiotic stresses and chemical treatments will be presented. Although this work shows expression profiling at the sub-genomic level, this could be a good starting point to understand the complexity of global defense mechanism in hot pepper.

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Characterization of Excision Repair Genes Related to Damaged DNA Repair from Eukaryotic Cells

  • Choi, In-Soon;Jin, Yong-Hwan;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
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    • v.17 no.1
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    • pp.1-6
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    • 1997
  • The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the partial cloning and characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the sequence homologous DNA to RAD4 gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. The level of the transcript did not increase upon UV-irradiation, suggesting that the RAD4 homologous gene in C. cinereus is not UV-inducible.

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Pyridoxine Deficiency on Neurotransmitters in the Developing Rat Brain - Catecholamine Metabolism- (Pyridoxine결핍이 뇌의 신경전달물질에 미치는 영향 - Catecholamine 대사 -)

  • Choi, Hay-Mie;Kang, Soon-Ah
    • Journal of Nutrition and Health
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    • v.17 no.3
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    • pp.199-209
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    • 1984
  • Pregnant rats were fed a pyridoxine deficient diet during the gestation and lactation. DEF I group received the deficient diet from delivery ; DEF II group, from the 15 th day of gestation. Body and brain weights, brain protein, DNA, RNA, plasma GOT and GPT, and catecholamines were measured. Effect of MAO inhibiting drug, pargyline, was determined. Brain protein, DNA, and RNA of offsprings of deficient groups were significantly lower than the control group, but RNA/ DNA, brain weight/DNA, and protein/DNA show that cell number were more affected than cell size by the pyridoxine deficiency during the 3rd week of gestation and lactation. Plasma GOT activities were more significantly different than plasma GPT between the control and deficient group. Brain norepinephrine of offsprings of deficient group were significantly lower than the control, but brain dopamine content was not significantly different from the control. At 2nd and 3rd week, norepinephrine was significantly depressed in deficient groups. Pargyline treatment affected a 1.2 fold increase in catecholamines in 3hr while the control had a 1.5 fold increase. Thus norepinephrine and dopamine synthesis was depressed in the deficient groups. Dopaminergic neurons may be less dependent on pyridoxine level than neurons from norepinephrine. Pyridoxine deficiency in maternal diet is not so critical to brain catecholamines of offspring except to the neonatal rats.

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Molecular Level Relationships of Purple Nonsulfur Bacteria and their Relatives

  • Lee, Sang-Seob;Yoon, Byoung-Su;Kim, Jae-Soo;Lee, Hyun-Soon
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.1-6
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    • 1994
  • DNA-DNA hybridization by kinetic method was carried out between species of purple nonsulfur photosynthetic bacteria and nonphotosynthetic bacteria. The degrees of homology percent were shown to be low (2-35 D%) with the exception of high homology % (72-88 D%) for strains within a species and between Rhodobacter capsulatus and Rhodopseudomonas blastica. The D% between the purple nonsulfur photosynthetic bacteria, Rhodopseudomonas palustris, and nonphotosynthetic bacteria, Pseudomonas aeruginosa ATCC 27853 or Bradyrhizobium japonicum were a little higher (26-33 D%) than the D% between any other photosynthetic bacteria. The homology % between Rhodopseudomonas blastica and Rhodobacter capsulatus was 72 D%, which showed genetic relationship.

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Molecular Cloning of Estrogen Receptor $\alpha$ in the Masu Salmon, Oncorhynchus masou

  • Sohn, Young Chang
    • Journal of Aquaculture
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    • v.17 no.1
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    • pp.62-68
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    • 2004
  • A cDNA encoding the masu salmon, Oncorhynchus masou, estrogen receptor $\alpha$ (msER$\alpha$) was cloned from the pituitary gland by polymerase chain reaction (PCR). This cDNA contains an open reading frame encoding 513 amino acid residues, and the calculated molecular weight of this protein is about 56,430 Dalton. The amino acid sequences of the DNA binding and ligand binding domains of msER$\alpha$ showed high homology to those of other fish species (84-100%). Reverse transcription PCR analysis showed that the mRNA level of msER$\alpha$ in the pituitary was slightly higher in estradiol-17$\beta$(E2) injected masu salmon than that of control fish. To test the biological activity of msER$\alpha$, the cDNA was ligated to a mammalian expression vector and transfected into a gonadotrope-derived cell line, L$\beta$T2, with a reporter plasmid including estrogen responsive element. Expression of the reporter protein, luciferase, was E2 and msER$\alpha$-dependent. The masu salmon ER$\alpha$ is structurally conserved among teleost species and functions as a transcriptional activator in the pituitary cells.

The Role of Mercury in the Etiology of Sperm Dysfunction in Holstein Bulls

  • Arabi, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.335-340
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    • 2006
  • A large number of toxicological substances and pharmacological and physical agents can cause reproductive intervention at the cellular and molecular level. The present study was designed to assess the effect of mercury ($HgCl_2$) at 50 to $550{\mu}M$ concentration ranges, in vitro, on the sperm membrane and DNA integrity, viability, and acrosomal status of normal bull spermatozoa. The samples were processed for sperm analyses using semen-diluting fluid (PBS, pH 7.2). We recorded a sharp increase in the lipid peroxidation/LPO rate; the highest was at $550{\mu}M$ mercury concentration, indicating a deleterious effect of mercury on the sperm membrane intactness. There was also a strong negative correlation between LPO rate and % viable spermatozoa (R = 0.987, p<0.001). Data obtained from a comet assay technique revealed that mercury is capable of inducing DNA breaks in the sperm nuclei. Interestingly, 92% of DNA breaks were double-stranded. The correlation between LPO rate and % DNA breaks was 0.984. Performing the gelatin test indicates that mercury is able to alter the integrity of acrosomal membranes showing an abnormal acrosome reaction. In this regard, a strong link was found between LPO rate and % halos (R = 0.990, p<0.001). Collectively, mercury proved to be a potent oxidant in the category of environmental factors affecting bull spermatozoa. Hence, considering the wide spread use of mercury and its compounds, these metals should be regarded with more concern.