• 제목/요약/키워드: Hybridization mechanisms

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Expression of the Multidrug Resistance Gene and its Product in Osteosarcomas of the Bone - Immunohistochemistry and In Situ Hybridization -

  • Park, Hye-Rim;Park, Yong-Koo
    • 대한골관절종양학회지
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    • 제3권1호
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    • pp.9-17
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    • 1997
  • Resistance to combination chemotherapy remains challenge in the treatment of osteosarcoma. One of the mechanisms of multiple drug resistance is an increased expression of the multidrug resistance gene(mdr1). Expression of the P-glycoprotein(mdr-1 gene product) was studied immunohistochemically and the mdr-1 gene by in situ hybridization in 33 osteosarcomas relating to various prognostic factors. Thirty cases out of 33 osteosarcomas(90.9%) showed positive cytoplasmic reactions with P-glycoprotein and nineteen instances(57.6%) were strong positive(2+). The older(>20 years) and female patients revealed more intense immunohistochemical reactions rather than those of the younger and male patients. Osteoblastic and chondroblastic osteosarcomas revealed more strong immunohistochemical reactions compared to fibroblastic types. There were no significant staining differences between the type of bony involvement, Broder's grade and the presence of necrosis. On follow-up, the mean survival rate was decreased in the strong positive group, however, this was not statistically significant. In situ hybridization for mdr-1 gene revealed positive signals in 22 cases out of 29 osteosarcomas(75.9%). Chemotherapy was done in 15 cases out of 28 patients(53.6%). The results of immunohistochemistry and in situ hybridization were not correlated with the protocols for chemotherapy. However, this result should be confirmed by a larger scale study about mdr1 mRNA expression.

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Single-molecule fluorescence in situ hybridization: Quantitative imaging of single RNA molecules

  • Kwon, Sunjong
    • BMB Reports
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    • 제46권2호
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    • pp.65-72
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    • 2013
  • In situ detection of RNAs is becoming increasingly important for analysis of gene expression within and between intact cells in tissues. International genomics efforts are now cataloging patterns of RNA transcription that play roles in cell function, differentiation, and disease formation, and they are demon-strating the importance of coding and noncoding RNA transcripts in these processes. However, these techniques typically provide ensemble averages of transcription across many cells. In situ hybridization-based analysis methods complement these studies by providing information about how expression levels change between cells within normal and diseased tissues, and they provide information about the localization of transcripts within cells, which is important in understanding mechanisms of gene regulation. Multi-color, single-molecule fluorescence in situ hybridization (smFISH) is particularly useful since it enables analysis of several different transcripts simultaneously. Combining smFISH with immunofluorescent protein detection provides additional information about the association between transcription level, cellular localization, and protein expression in individual cells.

Heterologous Microarray Hybridization Used for Differential Gene Expression Profiling in Benzo[a]pyrene-exposed Marine Medaka

  • Woo, Seon-Ock;Won, Hyo-Kyoung;Jeon, Hye-Young;Kim, Bo-Ra;Lee, Taek-Kyun;Park, Hong-Seog;Yum, Seung-Shic
    • Molecular & Cellular Toxicology
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    • 제5권4호
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    • pp.283-290
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    • 2009
  • Differential gene expression profiling was performed in the hepatic tissue of marine medaka fish (Oryzias javanicus) after exposure to benzo[a]pyrene (BaP), a polycyclic aromatic hydrocarbon (PAH), by heterologous hybridization using a medaka cDNA microarray. Thirty-eight differentially expressed candidate genes, of which 23 were induced and 15 repressed (P<0.01), were identified and found to be associated with cell cycle, development, endocrine/reproduction, immune, metabolism, nucleic acid/protein binding, signal transduction, or non-categorized. The presumptive physiological changes induced by BaP exposure were identified after considering the biological function of each gene candidate. The results obtained in this study will allow future studies to assess the molecular mechanisms of BaP toxicity and the development of a systems biology approach to the stress biology of organic chemicals.

Reproductive Isolation between Moroco oxycephalus and M. lagowskii (Pisces; Cyprinidae) in Korea

  • Kang, Young-Jin;Min, Mi-Sook;Yang, Suh-Yung
    • Animal cells and systems
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    • 제4권2호
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    • pp.109-115
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    • 2000
  • To clarify taxonomic status of the two sibling species, Moroco oxycephalus and M. lagowskii reproductive isolation mechanisms were investigated at sympatric area located in Kansung-up, Kosung-gun, Kangwon-do, Korea. Genetic analysis was performed to reveal mating system and intensity of Hybridization between the two species. The frequencies of hybrids were increased since 1989, and then the observed hybrid frequencies ($H_O$) did not significantly differ from the expected hybrid ($H_E$) in 1998 and 1999. However, based on histological analysis of two parents and their hybrid s gonads, the hybridizations between M. oxycephalus and M. lagowskii produced mostly fertile females but sterile males in accordance with Haldane s rule. Although it was suspected that pre- and postmating isolation mechanisms were affected between the two species, M. oxycephalus and M. lagowskii seemed to be strongly isolated with microhabitat at sympatry until 1997. Since 1998, hybrid frequencies were increased by habitat disturbance. However, their hybrid frequencies would be reduced by postmating isolation mechanisms. Therefore, the two species are considered to be distinct species recently diverged.

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압전-마찰전기 복합 소재 기반의 고출력 에너지 하베스팅 기술 개발 리뷰 (Review on the Recent Advances in Composite Based Highoutput Piezo-Triboelectric Energy Harvesters)

  • ;박현제;손민균;이태형;강대준
    • 세라미스트
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    • 제23권1호
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    • pp.54-88
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    • 2020
  • Global effort has resulted in tremendous progress with energy harvesters that extract mechanical energy from ambient sources, convert it to electrical energy, and use it for systems such as wrist watches, mobile electronic devices, wireless sensor nodes, health monitoring, and biosensors. However, harvesting a single energy source only still pauses a great challenge in driving sustainable and maintenance-free monitoring and sensing devices. Over the last few years, research on high-performance mechanical energy harvesters at the micro and nanoscale has been directed toward the development of hybrid devices that either aim to harvest mechanical energy in addition to other types of energies simultaneously or to exploit multiple mechanisms to more effectively harvest mechanical energy. Herein, we appraise the rational designs for multiple energy harvesting, specifically state-of-the-art hybrid mechanical energy harvesters that employ multiple piezoelectric and triboelectric mechanisms to efficiently harvest mechanical energy. We identify the critical material parameters and device design criteria that lead to high-performance hybrid mechanical energy harvesters. Finally, we address the future perspectives and remaining challenges in the field.

Screening of Differentially Expressed Genes in Heterosigma akashiwo, a Red-Tide Causing Organism, Induced by Exposure to High Light

  • Ko, Young-Seok;Cho, Kyung-Je;Moon, Byoung-Yong
    • Journal of Photoscience
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    • 제8권3_4호
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    • pp.93-97
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    • 2001
  • Heterosigma akashiwo has been reported as red-tide causing phytoplankton in the Korean coastal area during summer when they are exposed to high light. It also shows photosynthetic adaptability to strong light during culture in the laboratory. On the basis of these observations, we tried to find out some genes specifically expressed in Heterosimga akashiwo during exposure to high light, assuming that they might have some resistant mechanisms associated with light adaptation. For this purpose, we carried out DD-PCR to detect differentially expressed mRNAs from cells that had been illuminated under high light for 3 days. We found eight cDNA clones that had been expressed specificically for high light. When they were further screened by reverse Northern hybridization, three of them were identified to be positive cDNA clones. When these cDNA fragments were subjected to DNA sequencing and then their base sequences were compared to GenBank database, one of them showed sequence homology 86% identical to the partial sequence of 16S rRNA gene of eubacterium CRO-18.

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Effect of Phytohormones and Chemical Inhibitors on Pathogenesis-related Genes Identified by Differential Hybridization in Rice Suspension Culture Cells

  • Kim, Sang-Gon;Wu, Jing-Ni;Wang, Yiming;White, Ethan E.;Choi, Young-Whan;Kim, Keun-Ki;Choi, In-Soo;Kim, Yong-Cheol;Kim, Sun-Hyung;Kang, Kyu-Young;Kim, Sun-Tae
    • The Plant Pathology Journal
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    • 제26권4호
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    • pp.386-393
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    • 2010
  • In order to study disease resistance mechanisms in rice against the rice blast fungus Magnaporthe grisea, we screened fungal elicitor-responsive genes from rice suspension-cultured cells treated with fungal elicitors employing differential hybridization (DH). By DH screening, 31 distinct rice clones were isolated and a majority of them were full-length cDNAs encoding pathogenesisrelated (PR) genes. Sixteen of the 31 genes were upregulated at 4, 8, and 12 h following fungal elicitor treatment. To elucidate the effect of signal molecules and biotic elicitors on the regulation of rice defense genes, we further characterized the transcriptional expression patterns of representative isolated PR genes; OsGlu1, OsGlu2, OsTLP, OsRLK, and OsPR-10, following treatment with fungal elicitor, phytohormones, cycloheximide, and inhibitors of protein phosphorylation. Jasmonic acid (JA) induced transcriptional expression of OsGlu1, OsTLP, and OsRLK, but not of OsGlu2 and OsPR-10 at any of the tested time points. Salicylic acid (SA) and abscisic acid weakly induced the expression of OsTLP and OsRLK. SA showed an antagonistic effect with fungal elicitor and JA. Cycloheximide suppressed all these genes upon elicitor treatment, except for OsGlu2. Staurosporine only induced the expression of OsRLK. Application of calyculin A strongly induced OsRLK expression, but suppressed the expression of OsGlu2. Our study yielded a number of PR genes that play a role in defense mechanisms against the rice blast fungus, as well as contribute towards the elucidation of crosstalk between phytohormones and other modifications during defense signaling.

Identification of Genes Involved in Primordial-primary Follicle Transition by Suppression Subtractive Hybridization

  • Park, Chang-Eun;Yoon, Se-Jin;Jeon, Eun-Hyun;Kim, Young-Hoon;Lee, Sook-Hwan;Lee, Kyung-Ah
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2002년도 국제심포지엄
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    • pp.98-98
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    • 2002
  • Recruitment of primordial follicles(PMF) is crucial for female fertility. however, factors and mechanisms that regulate this process is poorly understood. The present study was conducted to obtain an inclusive view of the gene expression and to identify novel factors and their pathways of regulating PMF arrest and/or growth initiation. Ovaries from one-day neonatal(consists of oocyte and PMF) and five-day old(consists of PMF and primary follicles, PRIF) mice were collected, either total RNA or mRNA was isolated, and suppression subtractive hybridization(SSH) was used to isolate and clone genes that differentially expressed in day 1 and day 5 ovaries. Confirmation that some of these genes are differentially expressed in PMF and/or in PRIF was accomplished by using laser captured microdissection(LCM), RT-PCR. in situ hybridization(ISH) and/or immunohistochemistry(IHC). In toto, 357 clones were sequenced and analyzed by BLAST and RIKEN program. Sequences of 330 clones significantly matched database entries while 27 clones were novel. Forty-two and 47 different genes were identified as differentially expressed in day 1 and day 5 ovaries, respectively, while 7 genes were expressed in both stages of ovaries. Day 5-subtracted library included several genes known as markers far growing follicles, such as ZP2, MATER, and fetuin. Among the genes with assigned functions, 23.8% was associated with cell cycle/apoptosis regulation, 7.1% with cellular structure, 11.9% with metabolism, 26.2% with signal transduction, and 31.0% with gene/protein expression in day 1; while 10.6%, 17.0%, 23.5%, 25.5%, and 23.4% in day 5, respectively. Genes such as GDF-8, Lats2, Septin2, and Weel were the highly expressed genes in PMF, while HSP84, Laminin2, MATER, MTi7, PTP, and Wrn were highly expressed genes in PRIF. We have successfully discovered list of genes expressed in day 1 and day 5 ovaries and confirmed that some of them are differentially expressed in PMF and/or PRIF. Gene expression profile from the present study would provide insight for the future study on the mechanism(s) involved in primordial-primary follicular transition. This work was Supported by Korean Health 21 RND Project, Ministry of Health and Welfare, Korea (01-PJ10-PG6-01GN13-0002).

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Functional Genomics in the Context of Biocatalysis and Biodegradation

  • Koh Sung-Cheol;Kim Byung-Hyuk
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2002년도 추계학술대회
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    • pp.3-14
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    • 2002
  • Functional genomics aims at uncovering useful information carried on genome sequences and at using it to understand the mechanisms of biological function. Elucidating the unknown biological functions of new genes based upon the genomics rationales will greatly speed up the extensive understanding of biocatalysis and biodegradation in biological world including microorganisms. DNA microarrays generate a system for the simultaneous measurement of the expression level of thousands of genes in a single hybridization assay. Their data mining (transcriptome) strategy has two categories: differential gene expression and coordinated gene expression. Furthermore, measurement of proteins (proteome) generates information on how the transcribed sequences end up as functional characteristics within the cell, and quantitation of metabolites yields information on how the functional proteins act to produce energy and process substrates (metabolome). Various composite functional genomics databases containing genetic, enzymatic and metabolic information have been developed and will contribute to the understanding of the life blue print and the new discoveries and practices in biocatalysis and biodegradation that could enrich their industrial and environmental applications.

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Isolation of Human CYP4F2 genomic DNA and its $5^I$ End Regulatory Region Structure

  • Jin, Hyung-Jong
    • Archives of Pharmacal Research
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    • 제21권1호
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    • pp.35-40
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    • 1998
  • Human cytochrome P450 4F2 shows high regioselectivity in hydroxylation of stearic acid and leukotriene $ B_4.$ As a first step of its regulation study, human cytochrome P450 4F2 genomic DNA was isolated from liver of a person who was administered clofibrate for 10 years. From Southern hybridization, restriction enzyme digestion and sequencing experiments, isolated genomic DNA fragment was found to contain around 32 Kb DNA and more than 20 Kb of $5^I$ end regulatory region. Sequences of the structural gene region revealed exon 1 and exon 2. Further regulation studies would elucidate the feedback mechanisms of the oxidative degradation of fatty acids, inflammatory response and the clearance of leukotriene B4 in the liver. Furthermore, regulation study of this gene could explain the species difference in responses to peroxisome proliferator and help in the safety evaluation of peroxisome proliferating chemicals to human being.

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