• Title/Summary/Keyword: Human DNA

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Growth inhibitory and antioxidative effects of crude methanolic extract from Euonymus alatus (Thunb.) Sieb on SKBR3 human breast cancer cell line (SKBR3 유방암세포주에 대한 귀전우 메탄올 추출물의 성장억제 및 항산화효과)

  • Park, Young-Soo;Han, Ji-Young;Lee, Tae-Kyun;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.18 no.1
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    • pp.45-54
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    • 2005
  • 이 연구는 SKBR3 인간 유방암세포주에 대한 귀전우 메탄올추출물(CME)의 증식억제 효과, 세포사 유발 효과 및 항산화 활성을 확인하기 위해 이루어졌다. SKBR3 유방암세포주는 48시간동안 다양한 농도($0{\sim}20{\mu}g/m{\ell}$)의 CME가 제공된 곳에서 배양되었고, MMT 측정법을 이용하여 세포생존율을 평가하였다. CME의 50%에서 효과를 나타내게 하는 약물농도인 $ED_{50}$ (effective dose 50%)은 $6.5{\pm}0.3{\mu}g/m{\ell}$) 이며, 투여량이 증가함에 따라 농도에 의존하여 세포증식이 억제되는 것으로 나타났다. 또한 CME의 증식억제 효과는 유방암세포주의 세포사와 관련됨의 세포의 형태학적 변화와 올리고뉴클레오솜 DNA 파편의 확인을 통해 알 수 있었다. 또한 다양한 농도와 배양시간에서 CME가 ROS의 생산을 억제한다는 것을 확인할 수 있었다. 이런 결과들은 귀전우의 메탄올추출물이 SKBR3 인간 유방암세포주에 대해 강력한 증식억제 효과와 강한 항산화 효과를 나타낼 뿐만 아니라 세포사를 유도하는 효능을 가지고 있음을 시사한다. 이러한 효능은 약물에 대한 노출시간과 투여량에 의존하였다. 따라서 귀전우는 다양한 기전에 의해 유방암 세포에 대한 억제효과를 가질 수 있을 것으로 인식할 수 있다.

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Antioxidant Activity and Protective Effects of 9-hydroxy-$\alpha$-tocopherone from Viola mandshurica Extracts (제비꽃(Viola mandshurica) 추출물로부터 분리된 9-hydroxy-$\alpha$-tocopherone의 항산화 활성 및 세포 보호효과)

  • Lee, Mi-Ra;Hwang, Ji-Hwan;Park, Jae-Hee;Kim, Hyun-Jung;Park, Eun-Ju;Park, Hae-Ryong
    • Korean Journal of Pharmacognosy
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    • v.41 no.3
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    • pp.166-173
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    • 2010
  • Oxidative stress to proteins, lipids, or DNA is higher in human autopsy tissue and in rodent models of a number of neurodegenerative conditions, including Alzheimer's and Parkinson's disease. On the basis of this information, we established a screening system using N18-RE-105 cells to identify therapeutic agents that can protect cells from glutamate toxicity. During the course of our screening program, we recently isolated the active compound 9-hydroxy-$\alpha$-tocopherone ($\alpha$-TP), which prevents glutamate-induced cell death, from Viola mandshurica. The chemical structure of $\alpha$-TP was identified using spectroscopic methods and by comparison with literature values. Antioxidant activity and protective effects of $\alpha$-TP were evaluated by DPPH radical-scavenging assay, morphological assay, MTT reduction assay, and lactate dehydrogenase (LDH) release assay. These results suggest that $\alpha$-TP could be a new potential chemotherapeutic agent against neuronal diseases.

Genetic Diversity among Local Populations of the Gold-spotted Pond Frog, Rana plancyi chosenica (Amphibia: Ranidae), Assessed by Mitochondrial Cytochrome b Gene and Control Region Sequences

  • Min, Mi-Sook;Park, Sun-Kyung;Che, Jing;Park, Dae-Sik;Lee, Hang
    • Animal Systematics, Evolution and Diversity
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    • v.24 no.1
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    • pp.25-32
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    • 2008
  • The Gold-spotted pond frog, Rana plancyi chosenica, designated as a vulnerable species by IUCN Red list. This species is a typical example facing local population threats and extinction due to human activities in South Korea. A strategic conservation plan for this endangered species is urgently needed. In order to provide information for future conservation planning, accurate information on the genetic diversity and taxonomic status is needed for the establishment of conservation units for this species. In this study, we used a molecular genetic approach using the mitochondrial cytochrome b gene and control region sequences to find the genetic diversity of gold-spotted pond frogs within South Korea. We sequenced the mitochondrial DNA cytochrome b gene and control region of 77 individuals from 11 populations in South Korea, and one from Chongqing, China. A total of 15 cytochrome b gene haplotypes and 34 control region haplotypes were identified from Korean gold-spotted pond frogs. Mean sequence diversity among Korean gold-spotted pond frogs was 0.31% (0.0-0.8%) and 0.51% (0.0-1.0%), respectively. Most Korean populations had at least one unique haplotype for each locus. The Taean, Ansan and Cheongwon populations had no haplotypes shared with other populations. There was a sequence divergence between Korean and Chinese gold-spotted pond frogs (1.3% for cyt b; 2.9% for control region). Analysis of genetic distances and phylogenetic trees based on both cytochrome b and control region sequences indicate that the Korean gold-spotted pond frog are genetically differentiated from those in China.

A Simple, Rapid, and Automatic Centrifugal Microfluidic System for Influenza A H1N1 Viral RNA Purification

  • Park, Byung Hyun;Jung, Jae Hwan;Oh, Seung Jun;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.277.1-277.1
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    • 2013
  • Molecular diagnostics consists of three processes, which are a sample pretreatment, a nucleic acid amplification, and an amplicon detection. Among three components, sample pretreatment is an important process in that it can increase the limit of detection by purifying nucleic acid in biological sample from contaminants that may interfere with the downstream genetic analysis such as nucleic acid amplification and detection. To achieve point-of-care virus detection system, the sample pretreatment process needs to be simple, rapid, and automatic. However, the commercial RNA extraction kits such as Rneasy (Qiagen) or MagnaPure (Roche) kit are highly labor-intensive and time-consuming due to numerous manual steps, and so it is not adequate for the on-site sample preparation. Herein, we have developed a rotary microfluidic system to extract and purify the RNA without necessity of external mechanical syringe pumps to allow flow control using microfluidic technology. We designed three reservoirs for sample, washing buffer, and elution buffer which were connected with different dimensional microfluidic channels. By controlling RPM, we could dispense a RNA sample solution, a washing buffer, and an elution buffer successively, so that the RNA was captured in the sol-gel solid phase, purified, and eluted in the downstream. Such a novel rotary sample preparation system eliminates some complicated hardwares and human intervention providing the opportunity to construct a fully integrated genetic analysis microsystem.

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Photoluminescent Graphene Oxide Microarray for Multiplex Heavy Metal Ion Analysis

  • Liu, Fei;Ha, Hyun Dong;Han, Dong Ju;Park, Min Su;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.281.2-281.2
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    • 2013
  • Since heavy metal ions included in water or food resources have critical effects on human health, highly sensitive, rapid and selective analysis for heavy metal detection has been extensively explored by means of electrochemical, optical and colorimetric methods. For example, quantum dots (QDs), such as semiconductor QDs, have received enormous attention due to extraordinary optical properties including high fluorescence intensity and its narrow emission peaks, and have been utilized for heavy metal ion detection. However, the semiconductor QDs have a drawback of serious toxicity derived from cadmium, lead and other lethal elements, thereby limiting its application in the environmental screening system. On the other hand, Graphene oxide (GO) has proven its superlative properties of biocompatibility, unique photoluminescence (PL), good quenching efficiency and facile surface modification. Recently, the size of GO was controlled to a few nanometers, enhancing its optical properties to be applied for biological or chemical sensors. Interestingly, the presence of various oxygenous functional groups of GO contributes to opening the band gap of graphene, resulting in a unique PL emission pattern, and the control of the sp2 domain in the sp3 matrix of GO can tune the PL intensity as well as the PL emission wavelength. Herein, we reported a photoluminescent GO array on which heavy metal ion-specific DNA aptamers were immobilized, and sensitive and multiplex heavy metal ion detection was performed utilizing fluorescence resonance energy transfer (FRET) between the photoluminescent monolayered GO and the captured metal ion.

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Identification of two cytopathogenic agents, Mycoplasma hyorhinis and mammalian orthoreovirus 3 based on modified particle associated nucleic acids PCR

  • Kim, Hye Kwon;Moon, Hyoung Joon;Park, Seong Jun;Rho, Se Mi;Han, Jae Yeon;Nguyen, Van Giap;Park, Bong Kyun
    • Korean Journal of Veterinary Research
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    • v.51 no.2
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    • pp.129-137
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    • 2011
  • Swine diseases could be caused by unrecognized or minor pathogens. In this study, two unknown cytopathogenic agents were isolated from swine, through cell culture. In order to identify these two cytopathogenic agent (designated CP129 and #2045-7), a particle associated nucleic acids PCR (PANPCR) from previous paper was used with simple modification. The cloning procedure was more specified in this study by adding cell control system. According to the modified PAN-PCR, two and four agentsspecific DNA sequences were obtained from CP129 and #2045-7, respectively, and they were identified as Mycoplasma (M.) hyorhinis and Mammalian orthoreovirus by nucleotide BLAST. Since M. hyorhinis (CP129) was filterable and non-visible by microscope, this unusual virus-like nature of M. hyorhinis (CP129) was discussed. Especially, the reovirus (#2045-7) was a serotype 3 and a triple reassortant among three serotypes of reoviruses. It was grouped with recently reported reoviruses from disease cases (swine, human and feline), based on the genetic analysis of L1 and S1 partial sequences. In conclusion, two unknown cytopathogenic agents were successfully identified using modified PAN-PCR with cell control system and they were characterized in this study.

Expression of Escherichia coli Heat-labile Enterotoxin B Subunit (LTB) in Saccharomyces cerevisiae

  • Rezaee Mohammad Ahangarzadeh;Rezaee Abbas;Moazzeni Seyed Mohammad;Salmanian Ali Hatef;Yasuda Yoko;Tochikubo Kunio;Pirayeh Shahin Najar;Arzanlou Mohsen
    • Journal of Microbiology
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    • v.43 no.4
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    • pp.354-360
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    • 2005
  • Heat-labile enterotoxin B subunit (LTB) of enterotoxigenic Escherichia coli (ETEC) is both a strong mucosal adjuvant and immunogen. It is a subunit vaccine candidate to be used against ETEC-induced diarrhea. It has already been expressed in several bacterial and plant systems. In order to construct yeast expressing vector for the LTB protein, the eltB gene encoding LTB was amplified from a human origin enterotoxigenic E. coli DNA by PCR. The expression plasmid pLTB83 was constructed by inserting the eltB gene into the pYES2 shuttle vector immediately downstream of the GAL1 promoter. The recombinant vector was transformed into S. cerevisiae and was then induced by galactose. The LTB protein was detected in the total soluble protein of the yeast by SDS-PAGE analysis. Quantitative ELISA showed that the maximum amount of LTB protein expressed in the yeast was approximately $1.9\%$ of the total soluble protein. Immunoblotting analysis showed the yeast-derived LTB protein was antigenically indistinguishable from bacterial LTB protein. Since the whole-recombinant yeast has been introduced as a new vaccine formulation the expression of LTB in S. cerevisiae can offer an inexpensive yet effective strategy to protect against ETEC, especially in developing countries where it is needed most.

Effect of Several Species of the Family Rubiacea on Cytotoxicity and Apoptosis in HL-60 cells

  • Ju Sung-Min;Lee Jun;Choi Ho-Seung;Kim Sung-Hoon;Jeon Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.1
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    • pp.187-192
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    • 2006
  • Herbal medicines have been utilized to treat a variety of diseases, including cancer. Several species of the family rubiaceae have been reported to have antitumor activity. In this study, we report the cytotoxicity and antitumor activity exhibited dy the methanol extracts prepared from Rubia radix (RRME), Uncaria gambir (UGME) and Oldenlandia diffusa (ODME) (family: Rubiaceae) against human promyleloid leukemia cell line, HL-60. The cytotoxicity of RRME (2~20 ${\mu}g/ml$), UGME (20~200 ${\mu}g/ml$) and ODME (20~200 ${\mu}g/ml$) were assessed dy the MTT reduction assay. IC50 values for RRME, UGME and ODME were 11.0, 99.5 and 106.1 ${\mu}g/ml$, respectively. When the HL-60 cells were treated with RRME (10 ${\mu}g/ml$), UGME (120 ${\mu}g/ml$) and ODME (140 ${\mu}g/ml$) for 24 h, several apoptotic characteristics such as DNA fragmentation and morphologic changes were observed. Furthermore, flow cytometric analysis was peformed to determine the percent of apoptotic cells. The poupulation of sub-G1 hypodiploid cells was increased 37.49% in RRME treatment, 12.49% in UGME treatment and 7.21% in ODME treatment compared with untreated control cells (2.64%). To further confirm apoptotic cell death, we assayed caspase-3, -8 and -9 activities in RRME, UGME and ODME-treated cells. After treatment of RRME, UGME and ODME for 12 h, caspase-3, -8 and -9 activities significantly increased.compared to untreated control cells. These results show that RRME, UGME and ODME induced apoptotic cell death in HL-60 cells and may have a possibility of potential antitumor activities.

PHOTOCHEMISTRY AND PHOTOBIOLOGY OF PSORALENS

  • Shim, Sang-Chul;Jeon, Young Hee;Kim, DongWon;Han, GyuSeok;Yoo, Dong Jin
    • Journal of Photoscience
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    • v.2 no.1
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    • pp.37-45
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    • 1995
  • INTRODUCTION : Psoralens are planar tricyclic furocoumarins present in numerous plants and fungi found throughout the world.' Naturally occurring and synthesized psoralen derivatives(see Figure 1) are photosensitizers of UVA especially from 320 nm to 400 nm, a range at which cellular nucleic acids and proteins are weakly absorbing if any at all. Because of their skinphotosensitizing properties, these compounds have been used in the photochemotherapy of psoriasis and vitiligo. However, undesirable side effects such as carcinoma development in hairless mice as well as possible liver damage from the use of 8-methoxypsoralen(8-MOP) have been reported. The other photobiological effects include inactivation of DNA viruses, killing and mutagenesis of bacteria, inhibition of tumor transmitting capacity of various cells, and hyperpigmentation on human and guinea pig skin. PUVA(psoralen+UVA) photochemotherapy is in fact thousands of years old, having been used in Egypt and India since B.C. 1200-2000. Photochemotherapy for a common disfiguring disease, vitiligo, was practiced in the ancient world by physicians and herbalists who used boiled extracts of the fruits of certain umbelliferous plants, e.g. Ammi majus Linnaeus in Egypt or the leguminous plants, Psoralea corylifolia L. in India. It was first described by Kuske in 19388 that photosensitization of skin by plants was related to the presence of psoralen. He identified natural psoralens in plants as photosensitizers and isolated bergapten(5methoxypsoralen) from the oil of bergamot. The scientific interest in photosensitizing psoralens, however, has grown considerably after the introduction into clinics of the psoralen photochemotherapy for the treatment of psoriasis and of other skin (abbreviation)

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Genetic counseling in Korean health care system (유전상담의 제도적인 고찰)

  • Kim, Hyon-J.
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.1-5
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    • 2007
  • Unprecedented amount of genetic information being generated from the result of Human Genome Project (HGP) and advances in genetic research is already forcing changes in the paradigm of health and disease. The ultimate goal of genetic medicine is to use genetic information and technology to develop new ways of treatment or even prevention of the disease on an individual level for 'personalized medicine'. Genetics is play ing an increasingly important role in the diagnosis, monitoring and management of common multifactorial diseases in addition to rare single-gene disorders. While wide range of genetic testing have provided benefits to patients and family, uncertainties surrounding test interpretation, the current lack of available medical options for the diseases, and risks for discrimination and social stigmatization may remain to be resolved. However an increasing number of genetic tests are becoming commercially available, including direct to consumer genetic testing, yet public is often unaw are of their clinical and social implications. The personal nature of information generated by a genetic test, its power to affect major life decisions and family members, and its potential misuse raise important ethical considerations. Therefore appropriate genetic counseling is needed for patient to be informed with the benefits, limitations and risks of genetic tests, prior to informed consent for the tests. Physician also should be familiar with the legal and ethical issues involved in genetic testing to tell patients how w ell a particular genetic risk factor relates with likelihood of disease, and be able to provide appropriate genetic counseling. Genetic counseling become a mandatory requirement as global standard for many genetic testing such as prenatal diagnosis, presymtomatic DNA diagnostic tests and cancer susceptibility gene test for familial cancer syndrome. In oder to meet the challenge of genetic medicine of 21 century in korean health care system, professional education program and certification board for medical genetics specialist including non-MD genetic counselors should be addressed by medical society and regulatory policy of national health insurance reimbursement for genetic counseling to be in place to promote the implementation of clinical genetic service including genetic counseling for proper genetic testing.

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