• Parasites, Hosts and Diseases
• /
• v.42 no.3
• /
• pp.145-148
• /
• 2004

We compared the DNA sequence difference of isolates of Clonorchis sinensis from one Korean (Kimhae) and two Chinese areas (Guangxi and Shenyang), The sequences of nuclear rDNA (18S, internal transcribed spacer 1 and 2: ITS1 and ITS2) and mitochondrial DNA (cytochrome c oxidase subunit 1: cox1) were compared. A very few intraspecific nucleotide substitution of the 18S, ITS1, ITS2 and cox1 was found among three isolates of C. sinensis and a few nucleotide insertion and deletion of ITS1 were detected. The 18S, ITS1, ITS2 and cox1 sequences were highly conserved among three isolates. These findings indicated that the Korean and two Chinese isolates are similar at the DNA sequence level.

• Nutrition Research and Practice
• /
• v.11 no.2
• /
• pp.105-113
• /
• 2017

BACKGROUND/OBJECTIVES: A high-fat diet (HFD) induces obesity, which is a major risk factor for cardiovascular disease and cancer, while a calorie-restricted diet can extend life span by reducing the risk of these diseases. It is known that health effects of diet are partially conveyed through epigenetic mechanism including DNA methylation. In this study, we investigated the genome-wide hepatic DNA methylation to identify the epigenetic effects of HFD-induced obesity. MATERIALS AND METHODS: Seven-week-old male C57BL/6 mice were fed control diet (CD), calorie-restricted control diet (CRCD), or HFD for 16 weeks (after one week of acclimation to the control diet). Food intake, body weight, and liver weight were measured. Hepatic triacylglycerol and cholesterol levels were determined using enzymatic colorimetric methods. Changes in genome-wide DNA methylation were determined by a DNA methylation microarray method combined with methylated DNA immunoprecipitation. The level of transcription of individual genes was measured by real-time PCR. RESULTS: The DNA methylation statuses of genes in biological networks related to lipid metabolism and hepatic steatosis were influenced by HFD-induced obesity. In HFD group, a proinflammatory Casp1 (Caspase 1) gene had hypomethylated CpG sites at the 1.5-kb upstream region of its transcription start site (TSS), and its mRNA level was higher compared with that in CD group. Additionally, an energy metabolism-associated gene Ndufb9 (NADH dehydrogenase 1 beta subcomplex 9) in HFD group had hypermethylated CpG sites at the 2.6-kb downstream region of its TSS, and its mRNA level was lower compared with that in CRCD group. CONCLUSIONS: HFD alters DNA methylation profiles in genes associated with liver lipid metabolism and hepatic steatosis. The methylation statuses of Casp1 and Ndufb9 were particularly influenced by the HFD. The expression of these genes in HFD differed significantly compared with CD and CRCD, respectively, suggesting that the expressions of Casp1 and Ndufb9 in liver were regulated by their methylation statuses.

• Journal of Nutrition and Health
• /
• v.31 no.7
• /
• pp.1100-1111
• /
• 1998

The incorporation of docosahexaenoic acid(DHA) and arachidonic acid(AA) into brain and liver lipid has been compared in male pups from binth to 10 weeks old by feeding DHA-rich experimental diets or chow diets to dams from pregnancy in rats. The experimental DHA-rich diets contained 7g fish oil and 3g corn oil per 100g diet. There were three experimental groups, FO-I : Dams were fed DHA-rich diet during pregnancy and lactation, and their it pups fed the same diet until 10 weeks old. FO-II Dams fed chow diet during pregnancy and DHA-diet during lactation, and their pups fed the same DHA-diet until 10 weeks. FO-III : Dams fed chow diet during gestation and lactation, and then the pups fed DHA-diet after weaning. The relative % of DHA in hepatic lipid was about 12% with chow diets, but increased rapidly to 20-25% level when DHA-rich diets were supplied after weaning. The AA(%) of FO-III group was relatively high when a chow diet containing higher amount of linoleic acid was given, but there was no significant difference between the groups after feeding on a DHA-rich diet. When the DHA-rich diet was supplied from pregnancy(FO-I), the relative % of DHA in brain lipid was 13.7% at birth and continuously increased to a maximum level(17.2%) at 3-weeks and then was sustained until 5 weeks old. Similar levels of DHA incorporation were observed when DHA-rich diet was supplied from lactation(FO-II). However, the pups of FO-III group showed significantly lower levels of DHA incorporation(72%) at birth. These livels slowly increased and reached an 87% level of FO-I at 10 weeks when the pups ate DHA-rich diets after weaning. The relative % of AA in brain lipid was 10.4% in the FO-I group at birth, which was significantly lower than those of other groups, but there was no significant difference between groups after feeding DHA-rich diets in all groups. The Ah(%) level increased to maximum(11-12%) at 3-weeks and then was slightly reduced and was sustained at about 10% after S-weeks. Total amounts of DNA in the whole brain rapidly reached maximum level at 3-weeks and then was sustained at a constant level after S-weeks. DNA content was not significantly different between groups at birth, but it was significantly higher in FO-I and FO-II groups than in FO-III group at 3-weeks. However, DNA content in FO-III group was continuously increased to 80% level of FO-I at 10-weeks after feeding DHA-rich diet since weaning. In conclusion, the DHA(%) in whole brain was most effectively deposited when DHA-rich diet had been supplied during pregnancy and lactation in rats. However, DHA supplementation after weaning also improved the incorporaton of DHA into brain and content of DNA even though brain development was almost completed, which suggests that DHA supplementation might be necessary to improve brain development in humans during infancy as well as pregnancy and lactation. (Korean J Nutrition 31(7) 1100-1111, 1998)

• Journal of Nutrition and Health
• /
• v.39 no.8
• /
• pp.773-785
• /
• 2006

Vitamin E in the body system plays an important role in preventing chronic diseases by decreasing the oxidative stress by free-radicals. However, there are not enough researches on analyzing the primary factors affecting vitamin E levels in the blood in Korean adults. Therefore, the purpose of this research was to examine blood tocopherol levels and the primary factors affecting the status. A complete lifestyle survey was performed on 314 Korean adult men and surveyed their smoking, drinking and exercising habits. The average plasma level of ${\alpha}-\;and\;{\gamma}-tocopherol$ showed similar mutual relations with plasma total cholesterol (TC), triglyceride (TG), or low density lipoprotein cholesterol (LDL-C) levels (p<0.001). Plasma ${\alpha}-tocopherol$ level of the subjects did not show any difference as smoking, drinking and exercising habits changed. However, ${\gamma}-tocopherol$ per TG showed much lower figure in smokers than non smokers (p < 0.05). Amongst diet factors, plasma ${\alpha}-tocopherol$ level showed negative correlations with Vitamin E intake, while ${\gamma}-tocopherol$ level showed positive correlations with Vitamin E intake. Erythrocyte superoxide dismutase (SOD) activity and plasma tocopherol showed negative correlations, and catalase activity and plasma ${\alpha}-tocopherol$ showed positive correlationship. The level of cell DNA damage of Iymphocyte and plasma ${\alpha}-\;or\;{\gamma}-tocopherol$ showed negative correlations. As a result of this research, the factors that affect Korean adult men's plasma ${\alpha}-tocopherol$ level are plasma TG, LDL-C and cell DNA damage in Iymphocyte, while the factors that affect ${\gamma}-tocopherol$ level are plasma TG, LDL-C and vitamin E intake based on multiple regression analysis. These findings implies that the level of different types of tocopherol depends on slightly different factors. A further research is needed on the factors involved in the differentiation of the types of tocopherol.

• Development and Reproduction
• /
• v.21 no.3
• /
• pp.237-247
• /
• 2017

Mammalian spermatogenesis occurs in a precise and coordinated manner in the seminiferous tubules. One of the attempts to understand the detailed biological process during mammalian spermatogenesis at the molecular level has been to identify the testis specific genes followed by study of the testicular expression pattern of the genes. From the subtracted cDNA library of rat testis prepared using representational difference analysis (RDA) method, a complimentary DNA clone encoding type III member of a DnaJ family protein, DnaJC18, was cloned (GenBank Accession No. DQ158861). The full-length DnaJC18 cDNA has the longest open reading frame of 357 amino acids. Tissue and developmental Northern blot analysis revealed that the DnaJC18 gene was expressed specifically in testis and began to express from postnatal week 4 testis, respectively. In situ hybridization studies showed that DnaJC18 mRNA was expressed only during the maturation stages of late pachytene, round and elongated spermatids of adult rat testis. Western blot analysis with DnaJC18 antibody revealed that 41.2 kDa DnaJC18 protein was detected only in adult testis. Immunohistochemistry study further confirmed that DnaJC18 protein, was expressed in developing germ cells and the result was in concert with the in situ hybridization result. Confocal microscopy with GFP tagged DnaJC18 protein revealed that it was localized in the cytoplasm of cells. Taken together, these results suggested that testis specific DnaJC18, a member of the type III DnaJ protein family, might play a role during germ cell maturation in adult rat testis.

• Journal of Nutrition and Health
• /
• v.32 no.6
• /
• pp.654-660
• /
• 1999

Folate, a precursor of the coenzyme tetrahydrofolate, plays an important role in DNA replication and cell proliferation, and thus could influence rapidly proliferating immune cells such as leukocytes and splenocytes. The effects of dietary folate on folate concentrations of plasma, thymus, spleen and leukocytes were investigated in rats. The animals were raised for 6 weeks on semipurifed experimental diets containing 0mg, 2mg, 4mg, 8mg folate/kg diet. Folate concentrations were determined microbiologically using Lactobacillus casei(ATCC 7469), and DNA strand breaks produced by alkaline treatment were analyzed fluorometrically. When compared to folate adequate diet, the folate deficient diet(0mg folate/kg diet) resulted in lowest folate levels in plasma, thymus, spleen and leukocytes, and the highest DNA strand breaks in spleen cells and leukocytes. Dietary folate levels significantly increased folate concentrations of immune tissues, leukocytes, and the plasma in a dose dependent manner, folate concentrations being highest with a diet providing 8mg folate/kg diet. The percentages of the double strand DNA remaining in the splenocytes and leukocytes after alkaline treatment were significantly increased with higher amounts of dietary folate in a dose dependent manner. Folate intakes of 8mg than 4mg/kg diet was found to be more effective in the prevention of DNA strand breaks. The results of this study suggest that increased folate above the requirement level could improve DNA stabilities in immune cells.

• BMB Reports
• /
• v.46 no.9
• /
• pp.448-453
• /
• 2013

CpG-DNA has various immunomodulatory effects in dendritic cells, B cells, and macrophages. While induction of cytokines by CpG-DNA has been well documented in macrophages, the expression of costimulatory molecules in CpG-DNA treated macrophages has not yet been defined. Therefore, we investigated the effects of CpG-DNA on the expression of costimulatory molecules in RAW 264.7 cells. The surface expression of CD80 was slightly increased and CD83 expression was significantly increased in response to CpG-DNA. However, the expression of CD86 and MHC class II was not changed. As expression of CD83 mRNA was also increased by CpG-DNA, CD83 expression is regulated at a transcriptional level. To understand the contribution of signaling pathways to CD83 induction, we used pathway specific inhibitors. The NF-${\kappa}B$ inhibitor significantly reduced surface expression of CD83 as well as phagocytic activity of RAW 264.7 cells. Therefore, CD83 expression may contribute to the immunostimulatory effects of CpG-DNA in macrophage cells.

• Animal cells and systems
• /
• v.7 no.2
• /
• pp.159-164
• /
• 2003

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. The RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA and DNA-RNA helicase activities. To examine the extent of conservation of structure and function of a S. pombe RAD3 during eukaryotic evolution, the RAD3 homolog gene was isolated by screening of genomic DNA library. The isolated gene was designated as HRD3 (homolog of RAD3 gene). Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as HRD3 gene and this gene exists as a single copy in S. pombe. The transcript of 2.8 kb was detected by Northern blot analysis, The level of transcripts increased by ultraviolet (UV) irradiation, indicating that HRD3 is one of the UV-inducible genes in S. pombe. Furthermore, the predicted partial sequence of HRD3 protein has 60％ identity to S. cerevisiae RAD3 gene. This homology was particularly striking in the regions identified as being conserved in a group of DNA helicases. Gene deletion experiments indicate that the HRD3 gene is essential for viability and DNA repair function. These observations suggest evolutionary conservation of other protein components with which HRD3 might interact in mediating its DNA repair and viability functions.

• Reproductive and Developmental Biology
• /
• v.33 no.2
• /
• pp.93-98
• /
• 2009

In this study, bovine Dnmt1 cDNA was sequenced and detected Dnmt1 mRNA level in bovine tissues by northern blot, methylation pattern of genome by southern blot, specific localization of Dnmt1 in mouse and bovine preimplantation embryos by immunocytostaining and Dnmt1 protein level in ovary and testis by western blot. Bovine Dnmt1 cDNA sequence showed more homology with that of human than mouse and rat. The RNA level of Dnmt1 was 10 times higher expression in placenta than other tissues. This indicates that placenta was hypermethylated compared to others organs. The genomic DNA could not be cut by a specific restriction enzyme (HpaII) in placenta, lung and liver of bovine. It suggests that Dnmt1 in some somatic cells was already methylated. Dnmt1, which has the antibody epitope 1316~1616, was distributed in nucleus and cytoplasm including the stage of pronuclear stage and maturation of oocyte and gradually weaken to blastocyst stage compare to negative. In addition, Dnmt1 was strongly expressed in tetraploid embryo and cloned 8-cell than IVF 8-cell. An aberrant pattern of DNA methylation in cloned embryo may be abnormal development of fetus, embryonic lethality and placenta dysfunction. The somatic specific band (190kDa) was appeared in ovary and testis, but oocyte specific band (175kDa) was not. Further investigations are necessary to understand the complex links between the methyltransferases and the transcriptional activity of genes in the cloned bovine tissues.

• Journal of Nutrition and Health
• /
• v.36 no.3
• /
• pp.255-261
• /
• 2003

Food irradiation has been steadily increasing in many countries in line with increasing international trade and concerns about naturally occurring harmful contaminants in food. Although irradiation provides an excellent safeguard for the consumer by destroying almost 100% of harmful bacteria, it is necessary to ensure the safety of irradiated foods. This study was performed to investigate the effect of an irradiated diet on lipid peroxidation in the plasma, liver, small intestinal mucosa, and lymphocyte DNA damage in mice. Eight-week old ICR mice were assigned to two groups to receive either non-irradiated or irradiated (10 kGy) diets containing 20.38% fish powder and 6.06% sesame seeds for 4 weeks. The resulting changes in the degrees of lipid peroxidation were evaluated based on the level of plasma and liver thiobarbituric acid reactive substance (TBARS), transmission electron micrograph of jejunal mucosa, and free radical-induced oxidative DNA damage in lymphocytes, as measured by alkaline comet assay (single cell gel electrophoresis). The peroxide values of the gamma irradiated diet were measured every week, and the sample for comet assay was taken at the end of the four week experimental period. There was no significant difference in food efficiency ratio between the two groups. The peroxide values of the diet were immediately increased to 35.5% after gamma irradiation and kept on increasing during storage. After 4 weeks, no differences in tissue or plasma TBARS value were observed between the two groups, but epithelial cells of jejumum showed osmiophillic laminated membranous structures, considered as myelin figures,. The oxidative DNA damage expressed as tail moment (TM) increased 30% in the blood lymphocytes of the mice fed the irradiated diet. In conclusion, the comet assay sensitively detected differences in lymphocyte DNA damage after feeding with the irradiated diet for 4 weeks. However, in order to ensure the safety of irradiated foods, it would be more useful to conduct a long-term feeding regimen using an irradiated diet and examine the level of lipid peroxidation and the state of oxidative stress in a greater range of organs.