• Title/Summary/Keyword: DNA screening

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Attenuation of Brain Injury by Water Extract of Goat's-beard (Aruncus dioicus) and Its Ethyl Acetate Fraction in a Rat Model of Ischemia-Reperfusion

  • Han, Hyung-Soo;Lee, Jong-Won
    • Preventive Nutrition and Food Science
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    • v.16 no.3
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    • pp.217-223
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    • 2011
  • Ischemic stroke constitutes about 80% of all stroke incidences. It is characterized by brain cell death in a region where cerebral arteries supplying blood are occluded. Under these ischemic conditions, apoptosis is responsible for the cell death, at least in part. Goat's-beard (Aruncus dioicus var. kamtschaticus) is a perennial plant that grows naturally in the alpine regions of Korea. In the present study, we first determined whether water extract of goat's-beard (HY1646) and some of its fractions prepared by partitioning with organic solvents could improve the viability of human hepatocellular carcinoma cells (HepG2) cultured under hypoxic condition by blocking apoptotic pathways. Based on the in vitro findings, we subsequently investigated whether HY1646 and the ethyl acetate fraction (EA) selected from cell culture-based screening could attenuate brain injury in a rat middle cerebral artery occlusion (MCAO) model of ischemia (2 hr), followed by 22 hours of reperfusion. The cell number was sustained close to that initially plated in the presence of HY1646 even after 24 hr of cell culture under hypoxic condition (3% $O_2$), at which time the cell number reached almost zero in the absence of HY1646. This improvement in cell viability was attributed to the delay in apoptosis, identified by the formation of DNA ladder in gel electrophoresis. Of fractions soluble in hexane, ethyl acetate (EA) and butanol, EA was chosen for the animal experiments because EA demonstrated the best cell viability at the lowest concentration (10 ${\mu}g$/mL). HY1646 (200 mg/kg) and EA (10 and 20 mg/kg) significantly reduced infarct size, an index of brain injury, by 16.6, 40.0 and 61.0%, respectively, as assessed by 2,3,5-triphenyl tetrazolium chloride staining. The findings suggest that prophylactic intake of goat's beard might be beneficial for preventing ischemic stroke.

Spontaneous canine T-cell lymphosarcoma in a Shih Tzu dog (시추 개에서 발생한 T-cell 유래 림프육종 증례)

  • Oh, Hong-Guen;Chung, Yung-Ho;Lee, Hyun-A;Hong, Sun-Hwa;Kim, Ok-Jin
    • Korean Journal of Veterinary Service
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    • v.34 no.4
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    • pp.403-407
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    • 2011
  • This study describes a canine lymphosarcoma with a rapid proliferation and recurrence. A 4-year-old, male, Shih Tzu dog was examined for acute swelling mass. The mass had been identified since 3 months ago and enlarged $10{\times}7$ cm and located in the right axillary region. The surgical removal was recommended when patient visited veterinarian and the operation was conducted. The removed tumor was $11{\times}8{\times}7$ cm and firm, lobulated and white cut surface. Routine screening laboratory test was assessed with blood and radiological analysis. The metastasis sign was not detected on thoracic and abdominal radiography. Blood test revealed decreased lymphocytes. After surgical removal of the mass, microscopic histopathological examination was performed to determine the final diagnosis. Histopathologically, the tumors are characterized by the same histological features, including the presence of neoplastic cellular populations, and lymphocytes infiltration in varying proportions. Also, DNA was extracted and PCR analysis was employed to analyze the origin of tumor cells. T-cell specific nucleic acid fragments were specifically amplified by PCR. On the basis of the laboratory results, the tumor was diagnosed with canine T-cell lymphosarcoma. On the basis of our knowledge, this is the first report of canine T-cell lymphosarcoma in a Shih Tzu dog.

Useful Biomarkers for the Diagnosis of Endometriosis (자궁내막증 진단을 위한 생표지자의 유용성)

  • Kim, Sun-Young;Lee, Byung-Seok
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.1-12
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    • 2010
  • 진단적 복강경을 하지 않으면 자궁내막증의 진단이 불가능하다는 점은 의사들이 해결해야 할 과제 중 하나이다. 아직까지는 자궁내막증을 진단할 수 있는 획기적인 표지자가 없기 때문에 CA-125 같은 종양 표지자의 혈중 농도를 측정하였으나 진단 도구로 이용하기에는 한계가 있다. 이러한 이유로 초기 자궁내막증을 진단할 수 있는 방법을 연구하기 위한 여러 시도들이 있었는데 특히 자궁내막증 1, 2기 환자에서 병의 초기 상태에 복강경적 치료를 하였을 경우 자연 임신 성공률이 2배 가까이 높은 것으로 보고되었기 때문에 불임 여성에 있어 자궁내막증의 진단 시기는 임상적으로도 그 중요성이 매우 크다고 할 수 있겠다. CA-125는 자궁내막증 환자의 추적관찰에 있어 특이도가 높은 편이며 효용성이 있는데 특히 수술적 치료 후 장기적으로 병의 활성 혹은 재발을 평가하는데 있어 유용하다. 무작위적인 임상 연구 결과 자궁내막증과 관련된 불임이나 통증은 수술적 치료시 분명한 이득이 있는 것으로 보고된 바84 자궁내막증은 적절한 진단과 치료가 중요한 질환이라는 점을 다시 한번 상기해야 한다. 또한 병의 진행에 따른 여러 면역학적인 변화들이 확인되면서 자궁내막증의 진단에 있어 면역학적 표지자의 중요성이 부각되고 있다. 그 중에서도 복막액이나 혈청 내 사이토카인은 진단 도구로서 그 가능성에 주목을 받고 있으며 이에 대한 대규모 연구가 추후 필요할 것으로 사료된다. 최근의 면역학적 발견과 DNA 기술 발전은 자궁내막증의 진단에 있어 핵심적인 screening 도구의 발견에 일조할 것이며 이러한 기술적 발전을 근간으로 하여 머지 않아 획기적인 표지자가 개발될 것으로 기대한다.

Bioprospecting Endophytic Fungi and Their Metabolites from Medicinal Tree Aegle marmelos in Western Ghats, India

  • Mani, Vellingiri Manon;Soundari, Arockiamjeyasundar Parimala Gnana;Karthiyaini, Damodharan;Preethi, Kathirvel
    • Mycobiology
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    • v.43 no.3
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    • pp.303-310
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    • 2015
  • The increasing emergence of lead drugs for the resistance produced by the pathogenic strains and arrival of new diseases have initiated the need for searching novel metabolites with best anticancer and antimicrobial properties than the existing one. With this view, the investigation was conducted for the isolation, identification, and biological evaluation of potential endophytic fungi of Aegle marmelos, a medicinal tree used for more than three decades, for curing various disorders. A total of 169 endophytic fungal strains obtained from sampling and among those 67 were pigmented strains. Upon antagonistic screening, five endophytic fungal strains exhibited antagonistic potentiality by inhibiting the pathogens. These five potent strains were characterized at molecular level by sequencing the amplified internal transcribed spacer (ITS) 1 and ITS 4 regions of rDNA and they were grouped under order Pleosporales, Eurotiales, and Capnodiales. The metabolites from the respective strains were produced in fungal culturing media and extracted using polar solvents. Further, the extracts of five endophytes manifested antimicrobial activity against tested clinical pathogens and Alternaria alternata (FC39BY), Al. citrimacularis (FC8ABr), and Curvularia australiensis (FC2AP) exhibited significant antimicrobial profile against 9 of 12 tested pathogens, showing broad spectrum activity. The antioxidant levels of all the five endophytes revealed the highest activity at least concentrations, and major activity was unveiled by the members of order Pleosporales FC2AP and FC8ABr. This research explains the value of endophytic fungal extracts and its significance of antimicrobial and antioxidant properties.

The Detection of Genetically Modified Organisms in Soybean by DHPLC and Polymerase Chain Reaction (DHPLC와 중합효소연쇄반응에 의한 유전자재조합 콩의 검출)

  • Lee, Kyoung-Hae;Park, Su-Min
    • Food Science and Preservation
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    • v.15 no.1
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    • pp.88-93
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    • 2008
  • This paper focused on the detection of the genetically modified soybean (Glycine max L. MERRILL) samples to search for the speedy analysis methods. We have identified the PCR (polymerase chain reaction) assay with a newly developed technique called DHPLC (denaturing high performance liquid chromatography) to screen the GMO in soybean. The DHPLC is i1s ability to directly detection specific sequences of DNA by using column. With these characteristics. the DHPLC assay had the advantage of simplicity, rapidty could obtain result within 20 minutes. Whereas $15{\times}10^{-4}ng/{\mu}L$ concentration could be detected with the PCR analysis, $15{\times}10^{-5}ng/{\mu}L$ concentration could be detected with the DHPLC method. Therefore, DHPLC method was considered to be a simple, fast and sensitivity screening method rather than PCR analysis for GMO detection in soybean.

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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Development of a Single-nucleotide Polymorphism Marker for the Sw-5b Gene Conferring Disease Resistance to Tomato spotted wilt virus in Tomato

  • Lee, Hyung Jin;Kim, Boyoung;Bae, Chungyun;Kang, Won-Hee;Kang, Byoung-Cheorl;Yeam, Inhwa;Oh, Chang-Sik
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.730-736
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    • 2015
  • Tomato spotted wilt virus (TSWV) causes one of the most destructive viral diseases that threatens global tomato production. Sw-5b was reported as the resistance gene effective against TSWV. The objective of this research was to develop a single-nucleotide polymorphism (SNP) marker to distinguish tomato cultivars resistant to TSWV from susceptible cultivars for marker-assisted breeding. First, we determined genotypes for TSWV resistance in 32 commercial tomato cultivars using the previously reported Sw-5b gene-based marker. Then, DNA sequences of Sw-5b alleles in tomato cultivars showing resistant or susceptible genotypes were analyzed; a single SNP was found to distinguish tomato cultivars resistant to TSWV from susceptible cultivars. Based on the confirmed SNP, a SNP primer pair was designed. Using this new SNP sequence and high-resolution melting analysis, the same 32 tomato cultivars were screened. The results were perfectly correlated with those from screening with the Sw-5b gene-based marker. These results indicate that the SNP maker developed in this study will be useful for better tracking of resistance to TSWV in tomato breeding.

The Regulatory Effects of Low-Dose Ionizing Radiation on Ikaros-Autotaxin Interaction (저선량 방사선에 의한 Ikaros-Autotaxin 상호작용 조절 효과)

  • Kang, Hana;Cho, Seong-Jun;Kim, Sung Jin;Nam, Seon Young;Yang, Kwang Hee
    • Journal of Radiation Industry
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    • v.10 no.1
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    • pp.7-12
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    • 2016
  • Ikaros, a transcription factor containing zinc-finger motif, has known as a critical regulator of hematopoiesis in immune system. Ikaros protein modulates the transcription of target genes via binding to the regulatory elements of the genes promoters. However the regulatory function of Ikaros in other organelle except nuclear remains to be determined. This study explored radiation-induced modulatory function of Ikaros in cytoplasm. The results showed that Ikaros protein lost its DNA binding ability after LDIR (low-dose ionizing radiation) exposure. Cell fractionation and Western blot analysis showed that Ikaros protein was translocated into cytoplasm from nuclear by LDIR. This was confirmed by immunofluorescence assay. We identified Autotaxin as a novel protein which potentially interacts with Ikaros through in vitro protein-binding screening. Co-immunoprecipitation assay revealed that Ikaros and Autotaxin are able to bind each other. Autotaxin is a crucial enzyme generating lysophosphatidic acid (LPA), a phospholipid mediator, which has potential regulatory effects on immune cell growth and motility. Our results indicate that LDIR potentially regulates immune system via protein-protein interaction of Ikaros and Autotaxin.

Ginsenoside Rh2 upregulates long noncoding RNA STXBP5-AS1 to sponge microRNA-4425 in suppressing breast cancer cell proliferation

  • Park, Jae Eun;Kim, Hyeon Woo;Yun, Sung Hwan;Kim, Sun Jung
    • Journal of Ginseng Research
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    • v.45 no.6
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    • pp.754-762
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    • 2021
  • Background: Ginsenoside Rh2, a major saponin derivative in ginseng extract, is recognized for its anti-cancer activities. Compared to coding genes, studies on long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) that are regulated by Rh2 in cancer cells, especially on competitive endogenous RNA (ceRNA) are sparse. Methods: LncRNAs whose promoter DNA methylation level was significantly altered by Rh2 were screened from methylation array data. The effect of STXBP5-AS1, miR-4425, and RNF217 on the proliferation and apoptosis of MCF-7 breast cancer cells was monitored in the presence of Rh2 after deregulating the corresponding gene. The ceRNA relationship between STXBP5-AS1 and miR-4425 was examined by measuring the luciferase activity of a recombinant luciferase/STXBP5-AS1 plasmid construct in the presence of mimic miR-4425. Results: Inhibition of STXBP5-AS1 decreased apoptosis but stimulated growth of the MCF-7 cells, suggesting tumor-suppressive activity of the lncRNA. MiR-4425 was identified to have a binding site on STXBP5-AS1 and proven to be downregulated by STXBP5-AS1 as well as by Rh2. In contrast to STXBP5-AS1, miR-4425 showed pro-proliferation activity by inducing a decrease in apoptosis but increased growth of the MCF-7 cells. MiR-4425 decreased luciferase activity from the luciferase/STXBP5-AS1 construct by 26%. Screening the target genes of miR-4425 and Rh2 revealed that Rh2, STXBP5-AS1, and miR-4425 consistently regulated tumor suppressor RNF217 at both the RNA and protein level. Conclusion: LncRNA STXBP5-AS1 is upregulated by Rh2 via promoter hypomethylation and acts as a ceRNA, sponging the oncogenic miR-4425. Therefore, Rh2 controls the STXBP5-AS1/miR-4425/RNF217 axis to suppress breast cancer cell growth.

Comparison of Selection Efficiency between Marker-Assisted Selection and Phenotypic Selection for Development of Brown Planthopper Resistance Lines in Rice (벼멸구 저항성 계통선발을 위한 MAS의 선발효율 비교)

  • Kim, Suk-Man;Sohn, Jae-Keun
    • Korean Journal of Breeding Science
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    • v.40 no.1
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    • pp.48-53
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    • 2008
  • This study presents a case study designed to compare the selection efficiency between phenotypic selection (PS) and marker-assisted selection (MAS) in breeding of resistance lines to brown planthopper (BPH). The efficiency between PS and MAS were compared with four population such as the $F_2$, RILs ($F_6$), DH, and backcrosse ($BC_6F_5$) population, derived from a cross 'Samgang / Nagdong'. The resistance lines were selected using two markers, RM28493 and BpE18-3, related to BPH resistance were screened as resistance lines over 95% in PS. The costs required for BPH screening in the MAS system account for approximately 32% of the total costs of PS. The period needed to select the resistance plants was 30 days in PS and 7 days in MAS. Based on the results, we could establish the breeding system for selection of BPH resistance lines by MAS.