• Title/Summary/Keyword: tissue integration

Search Result 101, Processing Time 0.033 seconds

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

  • Min-cheol Kim;Tae-Hwan Jun
    • Proceedings of the Korean Society of Crop Science Conference
    • /
    • 2022.10a
    • /
    • pp.200-200
    • /
    • 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.

  • PDF

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

  • Franco, Rose-Ann;Nguyen, Thi Hiep;Lee, Byong-Taek
    • Proceedings of the Materials Research Society of Korea Conference
    • /
    • 2011.05a
    • /
    • pp.51.2-51.2
    • /
    • 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.

  • PDF

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

  • Lee, Seung-Min;Lee, Kwang-Ho
    • Journal of Biomedical Engineering Research
    • /
    • v.43 no.4
    • /
    • pp.219-229
    • /
    • 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 : Image Analysis and Visual Retrieval System for Biology Cell Image Bigdata (Visual Cell : 바이오세포 이미지 빅데이터를 위한 이미지 분석 및 시각적 검색 시스템)

  • Park, Beomjun;Jo, Sunhwa;Lee, Suan;Shin, Jiwoon;Yoo, Hyuk Sang;Kim, Jinho
    • The Journal of Bigdata
    • /
    • v.4 no.1
    • /
    • pp.53-61
    • /
    • 2019
  • The extracellular matrix, which provides the structural and biochemical support of surrounding cells, is a cell physiological modulator that controls cell division and differentiation. In the bio sector, the company produces Scapold, a three-dimensional support for tissue engineering, and cultivates stem cells in the produced Scapold to be transplanted into animals to assess tissue regeneration. This depends on components such as collagen in the tissue. Therefore, it is very important to identify the inclusion rate and distribution of components in the tissue, and the data are obtained by analyzing the color of the dyed tissue image. The process from image collection to analysis is costly, and the data collected and analyzed are managed in different formats by different research institutions. Therefore, data integration management and analysis results search are not being performed. In this paper, we establish a database that can manage relevant bigdata in an integrated manner, and propose a bio-image integrated management and retrieval system that can be searched based on color, an important analytical measure in this field of study.

  • PDF

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
    • /
    • v.5 no.2
    • /
    • pp.84-91
    • /
    • 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.

  • PDF

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
    • /
    • v.46 no.3
    • /
    • pp.204-213
    • /
    • 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
    • /
    • v.4 no.5
    • /
    • pp.297-304
    • /
    • 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.

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

  • Lee, Yun-Ho;Park, Joon-Bong;Kwon, Young-Hyuk;Herr, Yeek;Kim, Chong-Kwan
    • Journal of Periodontal and Implant Science
    • /
    • v.34 no.1
    • /
    • pp.163-175
    • /
    • 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.

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

  • 이정윤;이신우;박권우
    • Korean Journal of Plant Tissue Culture
    • /
    • v.25 no.2
    • /
    • pp.109-113
    • /
    • 1998
  • In this study we tried to transform an antimicrobial peptide gene (Shiva) under the promoter of tomato phenylalanine ammonia-lyase (tPAL5) into tobacco and potato plants. Antimicrobial peptide gene was isolated originally from giant silk moth (Hyalophora cecropia) and modified ie nucleotide sequence to increase antimicrobial activity. Transgenic tobacco plants were regenerated and their seeds were tested on the media containing kanamycin (500 mg/L). The results of PCR amplification and genomic Southern blot hybridization confirmed the integration of construct (tPAL5 promoter-Shiva-NOS-GUS-NOS) into chromosome. We observed that one of the transgenic tobacco plants showed chromosome rearrangement when integrated. In case of potato transformation, the efficiency of regeneration was maximized at the medium containing Zeatin 2mg/L, NAA 0.01mg/L, GA$_3$ 0.1mg/L. We also observed the high expression of GUS (${\beta}$-glucuronidase) enzyme which was located next to the terminator sequence of nopaline synthase gene (NOS) in the vascular tissue of stem, leaves of transgenic potatoes. This result suggested that a short sequence of Shiva gene (120 bp) and NOS terminator sequence might be served as a leader sequence of transcript when translated.

  • PDF

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

  • Shin, Dong-Hyun;Lee, Sei-Young;Koo, Bon-Seok;Kim, Se-Heon
    • Korean Journal of Head & Neck Oncology
    • /
    • v.25 no.1
    • /
    • pp.28-32
    • /
    • 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.