• 제목/요약/키워드: tissue integration

검색결과 101건 처리시간 0.029초

Gene Editing for Major Allergy Genes using Multiplex CRISPR-Cas9 System & Prime Editing in Peanuts (Arachis hypogaea L.)

  • Min-cheol Kim;Tae-Hwan Jun
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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    • pp.200-200
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    • 2022
  • Recently, food-induced allergies have emerged as major global concerns. In the past ten years, it has doubled in western nations, and it has also increased in Asia and Africa. In many cases of food allergy, peanut allergy is prevalent, typically permanent, and frequently life-threatening. Therefore, we utilized gene editing techniques on the three major allergen genes in peanuts, Ara h 1, Ara h 2, and Ara h 3. Using gibson assembly and golden gate assembly, we created two vectors, the gRNA-tRNA array CRISPR-Cas9 system and Prime-editing. Using LBA4404 strain and agrobacterium-mediated transformation, the vectors were transferred to two elite Korean peanut lines. After co-cultivation and tissue culture, we extracted the tissue cultured peanut DNA amplified the hygromycin resistance gene and Cas9 gene in the T-DNA region. The integration of the T-DNA region into the host genome was demonstrated by the presence of a specific band in some samples. There have only been a few reported peanut gene editing studies. So, this study will contribute to peanut allergy and gene editing research.

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Fabrication and Characterization of Novel Electrospun PVPA/PVA Nanofiber Matrix for Bone Tissue Engineering

  • Franco, Rose-Ann;Nguyen, Thi Hiep;Lee, Byong-Taek
    • 한국재료학회:학술대회논문집
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    • 한국재료학회 2011년도 춘계학술발표대회
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    • pp.51.2-51.2
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    • 2011
  • A novel electrospun nanofiber membrane was fabricated using combined poly (vinylphosphonic acid) (PVPA) and polyvinyl alcohol (PVA) intended for bone tissue engineering applications. PVPA is a proton-conducting polymer used as primer for bone implants and dental cements to prevent corrosion and brush abrasion. The phosphonate groups of PVPA have the ability to crosslink and attach itself to the hydroxyapatite surface facilitating faster integration of the biomaterial to the bone matrix. PVA was combined with PVPA to provide hydrophilicity, biocompatibility and improve its spinnability. To improve its mechanical strength, PVPA/PVA and neat PVA mixtures were combined to produce a multilayer scaffold. The physical and chemical properties of the of the fabricated matrix was investigated by SEM and TEM morphological analyses, tensile strength test, XRD, FT-IR spectra, swelling behavior and biodegradation rates, porosity and contact angle measurements. Biocompatibility was also examined in vitro by cytotoxicity and cell proliferation studies with MTT assay and cell adhesion behavior by SEM and confocal microscopy.

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의료용 소형 세라믹스 히터 소자의 개발 (Development of Micro-Ceramic Heater for Medical Application)

  • 이승민;이광호
    • 대한의용생체공학회:의공학회지
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    • 제43권4호
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    • pp.219-229
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    • 2022
  • In this study, we propose a miniaturized micro-ceramic heater device. After screen-printing a silver paste between pre-sintered two aluminum oxide plates to integrate a heating circuit, the device was fabricated through a low-temperature sintering process. In order to configure the optimal heating circuit integration condition, the output current evaluation and heating test were performed according to the number of screen prints of the silver paste at various voltages. A silver paste-based heating circuit printed with a line width of 200 ㎛ and a thickness of 60 ㎛ was successfully integrated on a pre-sintered alumina substrate through a low-temperature sintering process. In the case of the 5 times printed device, the thermal response showed a response rate of 18.19 ℃/sec. To demonstrate feasibility of the proposed device in the medical field, such as bio-tissue suturing and hemostasis, a voltage was applied to pig tissue in the device to test tissue change due to heat generated from the device. These results show the possibility that the proposed small ceramic heater could be used in the medical field based on its excellent temperature response.

Visual Cell : 바이오세포 이미지 빅데이터를 위한 이미지 분석 및 시각적 검색 시스템 (Visual Cell : Image Analysis and Visual Retrieval System for Biology Cell Image Bigdata)

  • 박범준;조선화;이수안;신지운;유혁상;김진호
    • 한국빅데이터학회지
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    • 제4권1호
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    • pp.53-61
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    • 2019
  • 주변 세포의 구조적, 생화학적 지지체를 제공하는 세포 외 기질은 세포의 분열과 분화 등을 좌우하는 세포생리 조절인자이다. 바이오 분야에서는 3차원 조직공학 지지체인 스캐폴드를 제작하고, 제작한 스캐폴드에 줄기세포를 배양해 동물에 이식해 조직 재생력을 평가한다. 이는 조직 내 콜라겐과 같은 구성성분에 좌우된다. 따라서 조직 내 구성성분의 포함율 및 분포를 파악하는 것이 매우 중요한데, 이에 관한 데이터를 염색된 조직 이미지의 색상을 분석함으로써 얻어낸다. 이때 이미지 수집부터 분석까지의 과정이 적지 않은 비용이 소모되고 있고, 수집되고 분석된 데이터를 연구 기관마다 상이한 포맷으로 관리하고 있다. 따라서 데이터 통합관리 및 분석결과 검색 등이 이루어지지 않고 있다. 본 논문에서는 관련 빅데이터를 통합적으로 관리할 수 있는 데이터베이스를 구축하고, 이 연구 분야에서 중요한 분석 척도인 색상을 기준으로 검색할 수 있는 바이오 이미지 통합 관리 및 검색 시스템을 제안한다.

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Permanent Mycoplasma Removal Removel from Tissue Culture Cells: A Genetic Approach

  • Motr, Gabriele;Preininger, Alexandra;Himmelspach, Michele;Plaimauer, Barbara;Arbesser, Christine;York, Heinz;Dorner, Friedrich;Schlokat, Use
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제5권2호
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    • pp.84-91
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    • 2000
  • Mycopasma contamination of tissue culture cells easily evades detection and, thus, represents a continous therat to cell biologists. In case where infected cell can not simply be replaced, attempts have to be made to eradicate mycoplacma from the tissue culture cells. A variety of anti-microbial agents have been shown to be toxic to mycoplasma strains ; however, cell associated mycoplasma are often protected from antibiotics at concentrations shown to be effective in vitro. Antibiotic concentrations high enough to be lethal to cell as sociated mycoplasmas frequently are also detrimentrations to the host cells, while moderately increased antibiotic levels tolerated by the host cells often lead to only temporary growth suppression and/or to the emergence of mycoplasma strains resistanct even to high concentrations of the antibiotis applied. Hare, a genetic approach for the elimination of mycoplasma from tissue culture cells that overcomes thens limitations is described. By expression of a selection marker conferring resistance to an otherwise toxic agent, Acholeplasma laidlawii infected BHK-21 cells used as the model system were enabled to temporarily tolerate antibiotic concentrations high enough to be lethal to cell associated mycopalsma while leaving the host cells unharmed. Upon successful mycoplasma eradicated, cultvation of the cured host cells in the absence of the selective agent yielded revertant cell clones that had regained susceptibillity to the toxic agent. Cressation of the selection marker expression was shown to result from the loss of the selection marker DNA, which is a consequence of the fact that the stable and permanent integration of foreign DNA in eucaryotic cell chrosomes is highly inefficient. Thus, the cells were cured from mycoplasma yet remained biochemically unaltered.

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A comparative study between sterile freeze-dried and sterile pre-hydrated acellular dermal matrix in tissue expander/implant breast reconstruction

  • Cheon, Jeong Hyun;Yoon, Eul Sik;Kim, Jin Woo;Park, Seung Ha;Lee, Byung Il
    • Archives of Plastic Surgery
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    • 제46권3호
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    • pp.204-213
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    • 2019
  • Background In implant-based breast reconstruction, acellular dermal matrix (ADM) is essential for supporting the inferolateral pole. Recent studies have compared non-sterilized freeze-dried ADM and sterilized pre-hydrated ADM, but have not assessed whether differences were attributable to factors related to sterile processing or packaging. This study was conducted to compare the clinical outcomes of breast reconstruction using two types of sterile-processed ADMs. Methods Through a retrospective chart review, we analyzed 77 consecutive patients (85 breasts) who underwent tissue expander/implant breast reconstruction with either freeze-dried ADM (35 breasts) or pre-hydrated ADM (50 breasts) from March 2016 to February 2018. Demographic variables, postoperative outcomes, and operative parameters were compared between freeze-dried and pre-hydrated ADM. Biopsy specimens were obtained for histologic analysis. Results We obtained results after adjusting for variables found to be significant in univariate analyses. The total complication rate for freeze-dried and pre-hydrated ADMs was 25.7% and 22.0%, respectively. Skin necrosis was significantly more frequent in the freeze-dried group than in the pre-hydrated group (8.6% vs. 4.0%, P=0.038). All other complications and operative parameters showed no significant differences. In the histologic analysis, collagen density, inflammation, and vascularity were higher in the pre-hydrated ADM group (P=0.042, P=0.006, P=0.005, respectively). Conclusions There are limited data comparing the outcomes of tissue expander/implant breast reconstruction using two types of sterile-processed ADMs. In this study, we found that using pre-hydrated ADM resulted in less skin necrosis and better integration into host tissue. Pre-hydrated ADM may therefore be preferable to freeze-dried ADM in terms of convenience and safety.

Test of a Multilayer Dose-Verification Gaseous Detector with Raster-Scan-Mode Proton Beams

  • Lee, Kyong Sei;Ahn, Sung Hwan;Han, Youngyih;Hong, Byungsik;Kim, Sang Yeol;Park, Sung Keun
    • IEIE Transactions on Smart Processing and Computing
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    • 제4권5호
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    • pp.297-304
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    • 2015
  • A multilayer gaseous detector has been developed for fast dose-verification measurements of raster-scan-mode therapeutic beams in particle therapy. The detector, which was constructed with eight thin parallel-plate ionization chambers (PPICs) and polymethyl methacrylate (PMMA) absorber plates, is closely tissue-equivalent in a beam's eye view. The gas-electron signals, collected on the strips and pad arrays of each PPIC, were amplified and processed with a continuous charge.integration mode. The detector was tested with 190-MeV raster-scan-mode beams that were provided by the Proton Therapy Facility at Samsung Medical Center, Seoul, South Korea. The detector responses of the PPICs for a 190-MeV raster-scan-mode proton beam agreed well with the dose data, measured using a 2D ionization chamber array (Octavius model, PTW). Furthermore, in this study it was confirmed that the detector simultaneously tracked the doses induced at the PPICs by the fast-oscillating beam, with a scanning speed of 2 m s-1. Thus, it is anticipated that the present detector, composed of thin PPICs and operating in charge.integration mode, will allow medical scientists to perform reliable fast dose-verification measurements for typical dynamic mode therapeutic beams.

천공형 티타늄막과 탈회동결건조골의 신생골 형성에 대한 영향 (The effect of new bone formation of titanium mesh and demineralized freeze-dried bone)

  • 이윤호;박준봉;권영혁;허익;김종관
    • Journal of Periodontal and Implant Science
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    • 제34권1호
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    • pp.163-175
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    • 2004
  • This study was performed to evaluate bone formation in the calvaria of rabbit by the concept of guided bone regeneration with titanium mesh membrane and demineralized freeze-dried bone. The animal was sacrificed at 2 weeks, 4 weeks, 8 weeks, and 12 weeks after the surgery. Non-decalcified specimens were processed for histologic analysis. 1. The titanium mesh but the biocompatibility was excellent the cell-occlusiveness was feeble. 2. The cell-occlusiveness was feeble and also the soft tissue growth of the upper part of the newly-formed bone after operating was excellent in early stage. 3. The maintenance ability of the space for the GBR very was excellent. 4. The titanium mesh the tissue-integration was superior the wound fixation ability excellent. 5. The demineralized freeze-dried bone did not promote the bone regeneration. 6. With the lapse of time, formation quantity of the bone some it increased, it increased quantity very it was feeble. Within the above results, the titanium mesh for the guided bone regeneration was excellent, the dεmineralized freeze-dried bone confirmed does not promote bone regeneration.

조직특이성 promoter를 이용한 Shiva 유전자의 식물체내 도입 (Introduction of Shiva Gene into tobacco and Potato Using Tissue-Specific Tomato PAL Promoter)

  • 이정윤;이신우;박권우
    • 식물조직배양학회지
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    • 제25권2호
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    • pp.109-113
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    • 1998
  • 본 연구에서는 상처, 병원균의 침입 등에 의하여 발현양이 크게 증폭되는 토마토 PAL유전자의 promoter에 giant silk moth(Hyalophora cecropia)로부터 분리한 lytic gene의 genetic code를 일부 변경한 Shiva 유전자를 부착하여 담배, 감자 등의 작물에 형질전환을 시도하였다. 형질전환된 담배로부터 얻은 종자에 대한 kanamycin저항성 유전자의 유전분석, PCR 증폭 혹은 genomic Southern blot hybridization에 의하여 tPAL5 promoter-Shiva fusion gene의 염색체내로의 integration을 확인하였다. Kanamycin 저항성 유전자의 유전분석에서 선발된 7개체를 PCR 분석 실시한 결과 모든 개체가 positive임이 확인되었으나, genomic Southern blot Hybridization으로는 4개체가 negative로 나타났다. 특히 한 개체의 경우는 chromosome rearrangement 현상이 일어난 것으로 추정되었다. 감자의 경우는 남작(Irish Cobbler) 품종이 Zeatin 2.0 mg/L NAA 0.01 mg/L, GA$_3$ 0.1mg/L을 포함한 배지에서 callus형성율 및 shooting율이 가장 높아서 재분화된 형질전환체를 얻을 수 있었다. 한편 GUS 유전자는 Shiva 유전자의 3' 말단에 존재하는 NOS terminator 때문에 translation까지의 발현이 어려울 것으로 예상되었으나 형질 전환하지 않은 담배에서보다 10배 이상의 GUS활성을 나타내었다. 또한 감자 조직에 X-gluc을 사용하여 GUS($\beta$-glucuronidase)의 기질로 작용하게 하여 효소활성 자리를 염색한 결과 줄기, 잎, 뿌리 등의 도관 조직에 다량 발혈됨을 확인할 수 있었다.

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두경부 편평세포암종의 발암 원인으로 인간유두종 바이러스(Human Papilloma Virus)의 역할 및 이와 관련된 발암 기전에 관한 연구 (The Etiologic Roles and Carcinogenic Mechanisms of Human Papilloma Virus in Head and Neck Squamous Cell Carcinoma)

  • 신동현;이세영;구본석;김세헌
    • 대한두경부종양학회지
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    • 제25권1호
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    • pp.28-32
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    • 2009
  • Background : The most frequently reported risk factors for head and neck suamous cell carcinoma are smoking and alcohol. But in a recent overview, human papilloma virus(HPV) infection was revealed the important carcinogenic factor in oropharyngeal cancer. We aimed to clarify whether HPV directly effects on the oncogenesis and biologic behavior of hean and neck squamous cell carcinoma by comparison with infection prevalence, and physical status of virus. Material and Method : We used HPV genotyping DNA chip(Biocore, Korea, Seoul) arrayed by multiple oligonucleotide probes of L1 sequence of 26 types of HPV and HPV genotypes are identified by fluorescence scanner. The copy numbers of HPV E2 and E6 open reading frames(ORF) were assessed using a TaqMan-based 5'-exonuclease quantitative real-time PCR assay. The ratio of E2 to E6 copy numbers was calculated to determine the physical status of HPV-16 viral gene. Results : We observed a significant difference in HPV prevalence between tonsillar cancer group and control group(73.1% vs. 11.6%), and most of the HPVs were type 16(87.2%) and integrated(94.1%) state. In terms of oral tongue cancer, we demonstrate that 30.5% has integrated HPV-16 in cancer tissue. But Glottic cancer only 1% is related to HPV-16 integration. Conclusion : This study revealed significant relationship of HPV prevalence with oropharyngeal and oral tongue squamous cell carcinoma. Most of HPV were 16 type and integrated or mixed, HPV-16 integration could be directly related to the carcinogenesis.