• Preventive Nutrition and Food Science
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• v.8 no.4
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• pp.390-394
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• 2003

In the post-genome period, the technique for identifying gene expression has been progressed to high throughput screening. In the field of molecular nutrition, the use of screening techniques to clarify molecular function of specific nutrients would be very advantageous. In this study, we have evaluated Zn-regulated gene expression in Zn-deficient, homocystein-treated EA.hy926 cells, using cDNA microarray, which can be used to screen the expression of many genes simultaneously. The information obtained can be used for preliminary assessment of molecular and signaling events modulated by Zn under pro-atherogenic conditions. EA.hy926 cells derived from human umbilical vein endothelial cells were cultured in Zn-adequate (control, 15 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) Dulbecco's MEM media under high homocysteine level (100 $\mu$M) for 3 days of post-confluency. Cells were harvested and RNA was extracted. Total RNA was reverse-transcribed and the synthesized cDNA was labeled with Cy3 or Cy5. Fluorescent labeled cDNA probe was applied to microarray slides for hybridization, and the slide was then scanned using a fluorescence scanner. The expression of seven genes was found to be significantly decreased, and one significantly increased, in response to treatment of EA.hy926 cells with Zn-deficient medium, compared with Zn-supplemented medium. The upregulated genes were oncogenes and tumor suppressor genes, cell cycle-related genes and transporter genes. The down-regulated gene was RelB, a component of the NF-kappaB complex of transcription factors. The results of this study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, namely Zn. Furthur study, using tailored-cDNA array and vascular endothelial cell lines, would be beneficial to clarify the molecular function of Zn in atherosclerosis, more in detail.

• The Journal of Korea Institute of Information, Electronics, and Communication Technology
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• v.15 no.5
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• pp.387-395
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• 2022

PDRN (Polydeoxyribonucleotide) is a DNA-derived polymer that promotes self-renewal of damaged cells and tissues as a tissue regeneration active material. PDRN is a DNA fragment cut into small sizes by various physical or chemical methods. When administered to the body, PDRN binds and stimulates the adenosine A2A receptor on the surface of tissue cells to promote cell regeneration, accelerate wound healing, and reduce pain. Although PDRN is prepared from testis or semen of fish in most cass, PDRN extraction from various plants species was performed in the present study. Among 7 tested plant species, the highest DNA yield and purity was obtained form mugwort (Chrysanthemum coronarium, C.c), followed by broccoli (Brassica oleracea, B.o). Then, we evaluated the in vitro wound healing capacity of PDRNs prepared from these two selected plants. PDRN from C.c and B.o. significantly stimulated the wound healing process at ㎍/ml range. The present study suggests that PDRN from plant species can be an effective alternative to PDRN from marine organism.

• Fisheries and Aquatic Sciences
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• v.26 no.9
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• pp.548-557
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• 2023

The purposes of this research were to identify fish species using DNA barcodes or partial sequences of cytochrome b (Cytb) and to assess the diversity of fish in the Mae Tam reservoir, Phayao province, Thailand. Fish samples were collected 3 times, during the winter, summer, and rainy seasons, from 2 sampling sites using gillnets with 3 mesh sizes (30, 50, and 70 mm). A total of 34 representative samples were classified into 12 species, 7 families and 6 orders by morphological- and DNA barcoding-based identifications. However, one cichlid species, Cichlasoma trimaculatum, could only be identified using DNA barcoding. Family Cyprinidae had the greatest diversity, 50.00%. The diversity, richness and evenness indices ranged from 0.43-0.65, 0.64-1.46, and 0.27-0.40, respectively, indicating that fish diversity at both sampling sites was relatively low. A comparison of the catch per unit effort (CPUE) with 3 different mesh sizes found that the 50 mm mesh size was the best (474.80 ± 171.56 g/100 m²/night), followed by the 70 mm (417.41 ± 176.24 g/100 m²/night) and 30 mm mesh sizes (327.88 ± 115.60 g/100 m²/night). These results indicate that DNA barcoding is a powerful tool for species identification. Our data can be used for planning the sustainable management of fisheries resources in the Mae Tam reservoir.

• Journal of Life Science
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• v.12 no.6
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• pp.715-720
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• 2002

A mutant Arabidopsis thaliana which is very sensitive to Ultraviolet-B(UV-B) radiation has been isolated by ethylmethane sulfonate(EMS) mutagenesis. Genetic cross proved the UV-sensitive gene(uvs) to segregate as a single Mendelian locus. For mapping of uvs, we crossed Arabidopsis thaliana Lansberg with uvs plant(Columbia), and made F2 plants by F1 selfcross. We designed 10 kinds of CAPS marker primers. Each primers amplifies a single mapped DNA sequence from uvs and Lansberg erecta ecotyres. Also identified was at least one restriction endonuclase for each of these PCR product that generates ecotype-specific digestion pattern. We got crossing over value of UB-sensitivity and each CAPS marker which located on different chromosome arm. The value of crossing over showed that uvs was linked to LFY3 which was on chromosome 5.

• Journal of Microbiology and Biotechnology
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• v.4 no.4
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• pp.256-263
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• 1994

The aceB gene, encoding for malate synthase, one of the key enzymes of glyoxylate bypass, was isolated from a pMT1-based Corynebacterium glutamicum gene library via complementation of an Escherichia coli aceB mutant on an acetate minimal medium. The aceB gene was closely linked to aceA, separated by 598 base pairs, and transcribed in divergent direction. The aceB expressed a protein product of Mr 83, 000 in Corynebacterium glutamicum which was unusually large compared with those of other malate synthases. A DNA-sequence analysis of the cloned DNA identified an open-reading frame of 2, 217 base pairs which encodes a protein with the molecular weight of 82, 311 comprising 739 aminoo acids. The putative protein product showed only limited amino acid-sequence homology to its counteliparts in other organisms. The N-terminal region of the protein, which shows no apparent homology with the known sequences of other malate synthases, appeared to be responsible for the protein s unusually large size. A potential calciumbinding domain of EF-hand structure found among eukaryotes was detected in the N-terminal region of the deduced protein.

• Journal of Periodontal and Implant Science
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• v.41 no.3
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• pp.157-163
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• 2011

Purpose: Curcumin is known to exert numerous biological effects including anti-inflammatory activity. In this study, we investigated the effects of curcumin on the production of interleukin-6 (IL-6) by murine macrophage-like RAW 264.7 cells stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a major cause of inflammatory periodontal disease, and sought to determine the underlying mechanisms of action. Methods: LPS was prepared from lyophilized P. intermedia ATCC 25611 cells by the standard hot phenol-water method. Culture supernatants were collected and assayed for IL-6. We used real-time polymerase chain reaction to detect IL-6 mRNA expression. $I{\kappa}B-{\alpha}$ degradation, nuclear translocation of NF-${\kappa}B$ subunits, and STAT1 phosphorylation were characterized via immunoblotting. DNA-binding of NF-${\kappa}B$ was also analyzed. Results: Curcumin strongly suppressed the production of IL-6 at both gene transcription and translation levels in P. intermedia LPS-activated RAW 264.7 cells. Curcumin did not inhibit the degradation of $I{\kappa}B-{\alpha}$ induced by P. intermedia LPS. Curcumin blocked NF-${\kappa}B$ signaling through the inhibition of nuclear translocation of NF-${\kappa}B$ p50 subunit. Curcumin also attenuated DNA binding activity of p50 and p65 subunits and suppressed STAT1 phosphorylation. Conclusions: Although further study is required to explore the detailed mechanism of action, curcumin may contribute to blockade of the host-destructive processes mediated by IL-6 and appears to have potential therapeutic values in the treatment of inflammatory periodontal disease.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.7
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• pp.3213-3222
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• 2016

Background: Methyl donor status influences DNA stability and DNA methylation although little is known about effects on DNA methyltransferases. The aim of this study was to determine whether methyl-donor status influences DNA methyltransferase (Dnmt) gene expression in cervical cancer cells, and if so, whether there are associated effects on global DNA methylation. Materials and Methods: The human cervical cancer cell line, C4-II, was grown in complete medium and medium depleted of folate (F-M+) and folate and methionine (F-M-). Growth rate, intracellular folate, intracellular methionine and homocysteine in the extracellular medium were measured to validate the cancer cell model of methyl donor depletion. Dnmt expression was measured by qRT-PCR using relative quantification and global DNA methylation was measured using a flow cytometric method. Results: Intracellular folate and methionine concentrations were significantly reduced after growth in depleted media. Growth rate was also reduced in response to methyl donor depletion. Extracellular homocysteine was raised compared with controls, indicating disturbance to the methyl cycle. Combined folate and methionine depletion led to a significant down-regulation of Dnmt3a and Dnmt3b; this was associated with an 18% reduction in global DNA methylation compared with controls. Effects of folate and methionine depletion on Dnmt3a and 3b expression were reversed by transferring depleted cells to complete medium. Conclusions: Methyl donor status can evidently influence expression of Dnmts in cervical cancer cells, which is associated with DNA global hypomethylation. Effects on Dnmt expression are reversible, suggesting reversible modulating effects of dietary methyl donor intake on gene expression, which may be relevant for cancer progression.

• Journal of Yeungnam Medical Science
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• v.12 no.2
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• pp.339-346
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• 1995

Assays for HBsAg, HBV DNA, anti-HBc and anti-HBs of 285 units of packed red blood cells supplied by Taegu Red Cross Blood Center were performed to evaluate the correlation between the prevalence of HBV DNA and the serologic markers for hepatitis B virus. None of 285 plasma samples was positive for HBsAg, however, HBV DNA were detected by polymerase chain reaction in 2 samples which both presented only with anti-HBc positivity. Of 204 samples tested for anti-HBs, 96 samples(47.1%) were positive and among 216 samples tested for anti-HBc, 80 samples(37.0%) were positive. Of 193 samples tested for both anti-HBs and anti-HBc, 80(41.1%) were all negative and 48(24.9%) were positive on both tests. Those samples which showed positivity only to anti-HBc were 25(13.0%). Considering the above results, transfusion-transmitted hepatitis B virus infection could be prevented by discarding anti-HBc positive blood, however, that may bring insufficient supply of donor bloods in the country like Korea where the prevalence of anti-HBc is high. Anti-HBc positive blood unequivocally positive for anti-HBs should be considered noninfectious for HBV and should be allowed to be transfused. It would reduce the amount of discarding donor blood as the routine blood donor screening tests presently used at Korea Red Cross Blood Center supplemented by anti-HBs and anti-HBc testing.

• Journal of Ginseng Research
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• v.14 no.2
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• pp.200-206
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• 1990

The effects of ginseng saponins, SRbl and G-Rc on the rat liver LDH A-gene transcriptional activity was investigated during prereplicative phase of rat liver after partial hepatectomy. Changes in LDH A-mRNA levels in regenerating rat liver after intraperitoneal administrations of G-Rbl or 'G-Rc were tested by slot blot hybridization methods. The results showed that G-Rbl (1 mg/100g B.W) and G-Rc (1 mg/100g B.W) caused marked increases of LDH A-mRNA contents by respectively 1.9- and 1.5-fold in rat liver at 5-hours after partial hepatectomy Dose dependent elect of G-Rbl and G-Rc (1-25 mg/ 100g B.W) on the LDH A-mRNA levels on regenerating rat liver were also analyzed. The maximal increases of liver LDH A-mRNA levels were observed with the doses of 1 mg for G-Rbl and 5 mg for G-Rc. However, when the administration doses of G-Rbl and G-Rc were increased to 20 mg, G-Rbl caused a marked decrease of LDH A-mRNA level to 61% of those in sham-operated rat liver. In contrast, G-Rc slightly decreased the liver LDH A-mRNA contents by 30% as compared to those of the maximum value but still maintained 22% higher LDH A-mRNA levels then those of sham-operated rate liver. On the basis of these experimental results, we conclude that ginseng saponin, G-Rbl and G-Rc have stimulatory effect at the lower concentration (1 mg/ 100g B.W) and inhibitory effect at the higher concentration (20 mg/ 100g B.W) on the LDH A-gene transcription during regeneration of rat liver. Additionally we also investigated the stimulatory effects of ginsenosides on the protein and DNA sinthetic activities in hepatocyte primary cell cultures isolated from regenerating rat liver. Both of G-Rc and -Re increased the synthetic rates of hepatocytes proteins and DNA at the administration doses of 50 us and 100 $\mu\textrm{g}$/3 ml/dish respectively representing 1.3-1.6 fold increases. From these results we postulate that G-Rc and -Re may have a mitogen ehincer activity for the hepatocyte proliferation during rat liver regeneration period.

• Journal of Life Science
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• v.27 no.4
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• pp.398-407
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• 2017

BTG 1 (B-cell translocation gene 1) gene was first identified as a translocation gene in a case of B-cell chronic lympocytic leukemia. BTG1 is a member of the BTG/TOB family with sharing a conserved N-terminal region, which shows anti-proliferation properties and is able to stimulate cell differentiation. In this study, we identified and characterized the pacific oyster Crassostrea gigas BTG1 (cg-BTG1) gene from the gill cDNA library by an Expressed Sequence Tag (EST) analysis and its nucleotide sequence was determined. The cg-BTG1 gene encodes a predicted protein of 182 amino acids with 57% 56% identities to its zebrafish and human counterparts, and is an intron-less gene, which was confirmed by PCR analysis of genomic DNA. Maximal homologies were shown in conserved Box A and B. The deduced amino acid sequence shares high identity with other BTG1 genes of human, rat, mouse and zebrafish. The phylogenic analysis and sequence comparison of cg-BTG1 with other BTG1 were found to be closely related to the BTG1 gene structure. In addition, the predicted promoter region and the different transcription-factor binding site like an activator protein-1 (AP-1) response element involved in negative regulation and serum response element (SRE) were able to be identified by the genomic DNA walking experiment. The quantitative real-time PCR analysis showed that the mRNA of cg-BTG1 gene was expressed in gill, heart, digestive gland, intestine, stomach and mantle. The cg-BTG1 gene was expressed mainly in heart and mantle.