• Korean Journal of Ichthyology
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• v.3 no.1
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• pp.48-57
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• 1991

Changes of the activities of the hepatic cells of female mud skipper, Boleophthalmus pectinirostris were investigated under transmission electron microscopy. Monthly changes of gonadosomatic index(GSI) and hepatosomatic index(HSI), variations of protein and nucleic acid contents(total RNA and DNA) of the liver tissues with the gonadal development phase were also studied. GSI began to increase from May(the growing stage), reaching the maximum value in late June(the mature stage), and then it began to decrease from late July(the degenerative stage), reaching the lowest value in late September. Monthly variations of HSI were negatively related to GSI. HSI decreased in the summer season when the ovary was getting mature and reached the maximum in mid October when the ovary was degenerating. In June(the mature stage), the female hepatic cells of the liver tissues became large and nuclei were hypertrophic. The amounts of glycogen particles and lipid droplets in the cells gradually decreased, while a number of granular endoplasmic reticulum increased. It was assumed that well-developed granular endoplasmic reticulum binding ribosomes are supposed to play the leading role in protein synthesis and deposition for vitellogenin in the cytoplasm. In July(the spawning period), glycogen particles and lipid droplets gradually increased, and then these substances were still observed in large quantity in August(the degenerative stage). The protein contents of the liver tissues with the gonadal phases of the ovaries were shown the maximum value($4.720{\pm}0.103\;mg/g$) in June, and afterwards gradually decreased being the minimum($3.640{\pm}0.130\;mg/g$) in July, and then gradually increased in August. The mean total RNA contents per gram of the liver tissues appeared the maximum($0.523{\pm}0.040\;mg/g$) in June, and afterwards gradually decreased to the minimum($0.158{\pm}0.006\;mg/g$) in July and slightly increased in August again. From these results, it could be assumed that protein contents were closely related to RNA contents. The mean total DNA contents per weight (gr) of the liver tissues appeared to be similar although there were some monthly fluctuations. The ratio of the mean total RNA/DNA were 0.745 in June, 0.262 in July, 0.341 in August respectively.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.112-112
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• 2003

Telomere란 진핵세포에 존재하는 DNA-protein 복합체로서 염색체의 말단부에 tandem repeated DNA 서열 (TTAGGG)과 특정 단백질로 구성되어 있으며 세포 분열이 진행함에 따라 이의 길이가 짧아지게 되고 일정 길이 이하가 되면 세포의 사망이 유발된다. 반면 telomerase는 ribonucleoprotein으로서 telomeric DNA의 합성에 관여하는 것으로 염색체의 말단에 telomeric DNA의 소실을 보충하는 역할로 알려져 있다. 최근 암, 노화 등과 관련하여 telomere 및 telomerase의 연구들이 활발히 진행되고 있으며, 다양한 세포들에 있어 이들의 존재와 역할에 대해서도 많은 연구들이 수행되고 있다. 포유동물의 초기 배자에 있어 telomere의 분포 양상과 telomerase의 activity의 분석은 배 발생의 기작과 배자의 세포적 특성을 구명하는데 매우 중요한 과제라 사료된다. 따라서 본 연구에서는 마우스의 초기 배 발생 단계별 수정란의 telomeric DNA의 분포 양상과 각 단계별 배자들의 telomerase activity를 제시하고자 하였다. 시험에 공시된 마우스는 4-6주령된 ICR계통으로 이들을 과배란 처리 후 자연 교배시켜 얻은 2-, 4-, 8-세포기배, 상실배 및 배반포배를 대상으로 하였다. Telomeric DNA의 양적 분석은 각 발생단계별 수정란의 표본을 제작하고 human telomere repeat probe를 이용하여 FISH (fluorescence in situ hybridization)를 시행하였으며, 분리된 할구들을 형광현미경으로 관찰 후 상을 포착하고 image analyser program (MataMorph, UIC, USA)을 이용하여 한 개의 세포내 telomere의 상대적 함량을 분석하였다. 발생 단계별 배자의 telomerase activity의 분석은 TRAP (telomeric repeat amplofication protocol) assay로 분석한 바 각 발생 단계별 30개의 수정란으로부터 핵 단백질을 추출하여 telomerase를 신장시키고 PCR을 시행한 후 15% PAGE gel loading하여 이의 activity를 확인하였다. 분석 결과, telomeric DNA의 함유율은 발생단계별 다소의 차이를 나타내었으며 telomerase activity는 모든 발생단계의 수정란에서 확인할 수 있었고, 특히 상실배부터 높게 나타남을 확인하였다.

• Korean Journal of Plant Resources
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• v.14 no.3
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• pp.235-240
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• 2001

Polymerase chain reaction (PCR) is used in a wide array of researches in plant molecular genetics and breeding. However, considerable time and cost are still required for the preparation of DNA suitable for reliable PCR results, especially in plant species containing high amounts of polyphenols. To reduce time and effort for PCR-based analysis, a simplified but reliable method was developed by a combinational employment of a simple and fast DNA extraction procedure and BLOTTO (Bovine Lacto Transfer Technique Optimizer) in reaction mixture. Genomic DNAs prepared by one-step extraction method from recalcitrant plant species such as Rubus coreanus, apple, grape and lettuce were successfully amplified by random primers in the reaction mixture containing 2 to 4% BLOTTO. Successful amplification of ${\gamma}$-TMT transgene in lettuce transformants by the specific primers was also achieved in the same condition, making rapid screening of positive transformants possible. Our results suggest that use of a simple DNA extraction procedure and incorporation of BLOTTO in reaction mixture in combination can reduce time and effort required for the analyses of a large number of germplasms and transformants by PCR-based techniques.

• Journal of Applied Biological Chemistry
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• v.59 no.3
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• pp.255-260
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• 2016

Reactive oxygen species (ROS) has been played a critical role in damage of DNA. Recently, many effort is focusing to develop the natural antioxidants for controlling ROS. Zelkova serrata, Ulmaceae, is close as plants which are planted in front of Korea villages. Although Zelkova serrata is familiar with Koreans, those of antioxidant activities and protective effects on oxidative DNA damage haven't studied. We demonstrated antioxidant activities and inhibitory effects on oxidative DNA damage of Leaf from Zelkova serrata with ethyl acetate fractions (EA) and hot water extracts (HW). Between the extracts, EA showed higher activities in 1,1-diphenyl-2-picryl-hydrazyl, 2,2'-azino-bis[3-ethylbenzthiazoline-6-sulphonic acid radical scavenging, $Fe^{2+}$ chelating and reducing power than HW. Also, those of total phenolic content are 56.63 and 51.61 mg/g respectively. In addition, ${\phi}X$-174 RF I plasmid DNA cleavage assay for inhibitory effect by oxidative DNA damage was both EA and HW has significant protective effect on oxidative DNA damage. The results suggested that leaf from Zelkova serrata with ethyl acetate fractions and hot water extracts have surpassing potential as natural resources with antioxidant and inhibitory effect on oxidative DNA damage.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.2
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• pp.149-155
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• 2011

The objective of this study was to evaluate the antioxidant and antigenotoxic activities of sansuyu fruit (Corni fructus, CF) at temperatures of $25^{\circ}C$, $50^{\circ}C$, and $90^{\circ}C$ using a water extraction method. Total phenolic content (TPC), DPPH radical-scavenging activity (RSA), and superoxide dismutase (SOD)-like activity, and ORAC (Oxygen radical absorbance capacity) values were determined. Also the antigenotoxicity of CF was determined by measuring inhibitory effects of $H_2O_2$ induced DNA damage in human leukocytes using the comet assay. The TPC in the CF extracts was 4.2, 4.6, and 5.5 g/100 g GAE in $25^{\circ}C$, $50^{\circ}C$, and $90^{\circ}C$, respectively. The DPPH RSA of the CF extracts increased in a dose-dependent manner over the range of $50\sim1000\;{\mu}g$/mL in all temperatures and the $SC_{50}$ of DPPH RSA of the CF extracts were not significantly different at different extraction temperatures. The $SC_{50}$ of SOD-like was the highest in CF extracted at $25^{\circ}C$ (1.1 mg/mL) followed by $90^{\circ}C$ (1.2 mg/mL) and $50^{\circ}C$ (1.3 mg/mL). The ORAC values of the CF extracts were not significantly different in low concentration ($10\;{\mu}M$/mL) and was in order of $25^{\circ}C$ ($5.7\;{\mu}M$ TE)< $90^{\circ}C$ ($6.2\;{\mu}M$ TE)< $50^{\circ}C$ ($8.5\;{\mu}M$ TE) in high concentration ($50\;{\mu}M$/mL). $200\;{\mu}M$ $H_2O_2$ induced DNA damages in human leukocytes were significantly reduced by the pretreatment with the CF extracts. These results suggest that sansuyu fruit (Corni fructus) can be used as a natural source for antioxidant activities and as antigenotoxic agents regardless of the water extraction temperature.

• The Korea Journal of Herbology
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• v.25 no.4
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• pp.55-59
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• 2010

Objective : This study was conducted to investigate the antioxidant activity and inhibitory effect on oxidative DNA damage of Nelumbinis Semen Extracts Methods : Nelumbins semen were extracted with hot-water and ethylacetate (EtOAC). The 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radical scavenging assay and $Fe^{2+}$ chelating assay were performed for antioxidative effect and ${\phi}X$-174 RF I DNA cleavage assay and intracellular DNA damage assay were used for inhibitory effect on intracellular DNA damage. Results : In DPPH, Hydroxyl radical scavenging activity and $Fe^{2+}$ chelating activity of EtOAC extracts were 96.22%, 53.53%, 64.72%, while those of hot-water extracts were 20.86%, 10.72%, 29.74% at $200{\mu}g/m{\ell}$, respectively. In ${\phi}X$-174 RF I plasmid DNA cleavage assay, the protective effects of EtOAC and hot-water extracts against oxidative DNA damage were 76% and 6% at $200{\mu}g/m{\ell}$, respectively. Conclusion : These results indicated that the seed extracts of Nelumbo nucifera can be used as a natural antioxidants, which effectively inhibits the oxidative DNA damage.

• Environmental Mutagens and Carcinogens
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• v.9 no.1
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• pp.1-12
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• 1989

To study the selective organospecific carcinogenesis by the specific chemical carcinogens, the breast cancer induction model by oral administration of 7, 12-dimethylbenzanthracene (DMBA) or by intravenous injection of N-methylni-trosourea (NMU) on female rats was analyzed. In the present experiment, we compared the effexts of ages on the chemical mammary carcinogenesis by studying the metabolic system of the carcinogenic activation, detoxification or DNA damage and repair. The breast tumor incidence was significantly higher in the young rats of 50 days old than in those of one year old rats. As an index of organospecific DNA damage or repair, the in vivo covalent binding index(CBI) of the specific organs by the specific chemical carcinogens was monitored. And for the analysis of carcinogenic activation, the quantity of cytochrome P450`s was determined with the respective type-specific monoclonal antibody, while the detoxication capacity was deduced by the activity monitoring of glutathione S-transferase (GST) and peroxidase. The skin tissues of the mammary region had the highest CBI with both of DMBA and NMU at 50 days of age. And there were contrasting differences in the contents of carcinogenic activation and detoxication system: that is, the content of T.C.D.D.-inducible cytochrome P450 was high, while the activities of GST and peroxidase was low in the mammary skin tissues at tumor prevalent age. These results led us to conclude that the molecular organospecific carcinogenesis, as illustrated with mammary carcinoge-nesis by DMBA and NMU, is operated probably through the differential capacity of the target tissues in the high carcinogenic activation, low detoxication and the low DNA repair function.

• Korean Journal of Plant Tissue Culture
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• v.25 no.5
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• pp.341-346
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• 1998

In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme dihydroflavonol 4-reductase (DFR) in Matthiola incana R. Br. A heterologous cDNA probe from Zea mays was used to isolate full-size DFR cDNA clone from a corolla-specific cDNA library. Comparison of the coding region of this DFR cDNA sequence including the sequences of Zea mays, Anthirrinum majus, Petunia hybrida, Callistephus chinensis, Dianthus caryophyllus and Rosa hybrida reveals a identity higher than 61% at the nucleotide level. The DFR transcript is G/C rich in monocotyledonous plants show a strong codon bias preferring codons with a G or C in the third position. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRNA from wild type and mutant plants and by in vitro expression yielding an enzymatically active reductase. Genomic southern blot analysis showed the presence of one gene for DFR in Matthiola incana. Northern blot analysis of the DFR wild type and mutant lines showed that the lack of DFR activity in the stable acyanic mutant k17b is clearly by a transcriptional block of the DFR gene.

• Korean Journal of Plant Resources
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• v.19 no.1
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• pp.161-168
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• 2006

To investigate the origin of backyeoncho (Opuntia ficus-indica var. saboten), we isolated 685 bp clone using ITS primer pairs. The rDNA consists of the genes coding for the partial 54 bp 185, 162 bp 5.8S, and partial 56 bp 26S. The coding regions are interrupted by two internal transcribed spacers, 193 bp ITS1 and 220 bp ITS2. The ITS2 of backnyeoncho in length was shorter than that previously registered in Cucurbitoideae plants. The GC contents was 66.8% in ITS1, and 67.7% in ITS2. The rDNA of backnyeoncho matched to the previously reported genes and showed a high similarity with the 95% identity with Pereskiopsis porteri (L708037). In the phylogenetic analysis, the backnyeoncho rDNA was clustered with Pereskiopsis porteri (L708037).

• Korean Journal of Plant Resources
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• v.37 no.2
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• pp.101-109
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• 2024

In this study, we investigated the antioxidant effects and active ingredients of Gynura Procumbens extracts obtained through various extraction methods for the development of natural-based cosmetics and pharmaceutical materials. The contents of compounds, total flavonoids, chlorogenic acid, and caffeic acid were compared at different concentrations, revealing the highest content of active ingredients in the 100% ethanol extract. Antioxidant activity assessed by DPPH and ABTS radical scavenging assays showed a concentration-dependent increase in antioxidant activity with the ethanol concentration. Additionally, we validated the DNA damage inhibition and anti-inflammatory effects of Gynura Procumbens extracts through UVB irradiation on Hs68 cell models. The 100% ethanol extract demonstrated significant inhibition of the expression of p-p53, γ-H2AX, iNOS, and COX-2 induced by UVB, indicating its potential in alleviating photodamage effects. Consequently, the efficient extraction of Gynura Procumbens for skin functional material development was confirmed, suggesting the suitability of ethanol or alcohol-based solvents.