• 제목/요약/키워드: Confocal laser scanning microscope

검색결과 165건 처리시간 0.021초

Characterization of Ha29, a Specific Gene for Helicoverpa armigera Single-nucleocapsid Nucleopolyhedrovirus

  • Guo, Zhong-Jian;An, Shi-Heng;Wang, Dun;Liu, Yan-He;Kumar, V. Shyam;Zhang, Chuan-Xi
    • BMB Reports
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    • 제38권3호
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    • pp.354-359
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    • 2005
  • Open reading frame 29 (ha29) is a gene specific for Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearSNPV). Sequence analyses showed that the transcription factor Tfb2 motif, bromodomain and Half-A-TPR (HAT) repeat were present at aa 66-82, 4-76, 55-90 of the Ha29 protein respectively. The product of Ha29 was detected in HearSNPV-infected HzAM1 cells at 3 h post-infection. Western blot analysis using a polyclonal antibody produced by immunizing a rabbit with purified GST-Ha29 fusion protein indicates that Ha29 is an early gene. The size of Ha29 product in infected HzAM1 cells was about 25 kDa, which was larger than the presumed size of 20.4 kDa. Tunicamycin treatment of HearSNPV-infected HzAM1 cells suggested that the Ha29 protein is N-glycosylated. Fluorescent confocal laser scanning microscope examination, and Western blot analysis of purified budded virus (BVs), occlusion-derived virus (ODVs), cell nuclear and cytoplasmic fraction, showed that the Ha29 protein was localized in the nucleus. Our results suggested that ha29 of HearSNPV encodes a non-structurally functional protein that may be associated with virus gene transcription in Helicoverpa hosts.

생물막 모델 시스템 (Biofilm modeling systems)

  • 김수경;이준희
    • 미생물학회지
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    • 제52권2호
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    • pp.125-139
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    • 2016
  • 생물막은 미생물들의 표면 부착 성장으로부터 유래된 복잡하게 구조화된 미생물들의 군집이다. 인간의 삶속에서 생물막은 다양한 감염을 매개하고 여러 사회, 문화, 산업 시설물들에서 많은 문제를 일으킨다. 생물막에 대한 이해가 과학자들에게 큰 관심을 끌고 있기는 하지만, 자연계에 존재하는 다양한 생물막을 직접 연구하는 것은 매우 어렵다. 따라서 다양한 연구실에서 생물막을 형성하기 위한 다양한 모델 시스템들이 제안되어 왔으며, 이들 모델에 기반한 많은 생물막 연구 방법들이 실제 생물막 연구에 쓰이고 있다. 이러한 생물막 모델들은 실제 환경속의 생물막들의 특징들을 모사하고 있지만 각기 나름대로의 장단점들을 가지고 있다. 본 리뷰에서는 현재 사용되고 있는 생물막 모델 시스템들을 소개하고, 그들의 장단점들을 설명하고자 한다.

Influence of size and insertion depth of irrigation needle on debris extrusion and sealer penetration

  • Uzunoglu-Ozyurek, Emel;Karaaslan, Hakan;Turker, Sevinc Aktemur;Ozcelik, Bahar
    • Restorative Dentistry and Endodontics
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    • 제43권1호
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    • pp.2.1-2.10
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    • 2018
  • Objectives: To determine the effect of size and insertion depth of irrigation needle on the amount of apical extruded debris and the amount of penetration depth of sealer using a confocal laser scanning microscope (CLSM). Materials and Methods: Twenty maxillary premolars were assigned to 2 groups (n = 10), according to the size of needle tip, 28 G or 30 G. Buccal roots of samples were irrigated with respective needle type inserted 1 mm short of the working length (WL), while palatal roots were irrigated with respective needle type inserted 3 mm short of the WL. Prepared teeth were removed from the pre-weighed Eppendorf tubes. Canals were filled with F3 gutta-percha cone and rhodamine B dye-labeled AH 26 sealer. Teeth were transversally sectioned at 1 and 3 mm levels from the apex and observed under a CLSM. Eppendorf tubes were incubated to evaporate the irrigant and were weighed again. The difference between pre- and post-weights was calculated, and statistical evaluation was performed. Results: Inserting needles closer to the apex and using needles with wider diameters were associated with significantly more debris extrusion (p < 0.05). The position of needles and level of sections had statistically significant effects on sealer penetration depth (p < 0.05 for both). Conclusions: Following preparation, inserting narrower needles compatible with the final apical diameter of the prepared root canal at 3 mm short of WL during final irrigation might prevent debris extrusion and improve sealer penetration in the apical third.

정상 위 조직에서 thymosin β4와 CD133의 발현 양상 분석 (Analysis of Expression Patterns of Thymosin β4 and CD133 in Normal Stomach)

  • 옥미선;차희재
    • 생명과학회지
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    • 제22권10호
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    • pp.1415-1419
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    • 2012
  • Thymosin ${\beta}4$ 는 대장암에서 암 줄기세포 마커인 CD133을 지닌 세포에서 지니지 않은 세포에 비해 강하게 발현된다고 보고되어 있다. 본 연구에서는 thymosin ${\beta}4$와 줄기세포 마커인 CD133의 상관관계를 정상 위 조직에서 관찰하였다. Thymosin ${\beta}4$와 CD133의 발현 양상은 tissue microarray 조직상에서 면역화학적 방법으로 관찰하였으며 thymosin ${\beta}4$와 CD133의 존재 위치는 면역형광 염색법 및 confocal microscope를 사용하여 조사하였다. Thymosin ${\beta}4$와 CD133은 동일한 양상으로 발현되었으며 모두 위의 선조직에서 강하게 발현되었다. 면역 형광 염색법으로 두가지 단백질을 동시에 염색한 결과 두 단백질이 동일한 위치에서 함께 존재하는 것으로 규명되었다. 이러한 결과는 thymosin ${\beta}4$와 CD133은 정상위의 선조직에서 발현되며 두 단백질의 발현 양상 및 위치가 동일하여 서로 긴밀한 상호작용을 할 가능성을 제시하고 있다.

공초점 레이저 주사 현미경을 이용한 우치 인공우식법랑질에서 불소도포 전·후의 재광화 효과에 대한 융합적 관찰 (The remineralization effect of topical fluoride agents using confocal laser scanning microscope on artificial enamel caries aspects of convergence observation)

  • 이민선;김효진
    • 한국융합학회논문지
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    • 제8권6호
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    • pp.139-145
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    • 2017
  • 본 연구는 우치의 인공우식법랑질에 대한 불소도포 후 재광화 효과를 공초점레이저주사현미경을 통해 비교하고자 진행되었다. 시편에 인공우식을 형성한 후, 대조군, 1.23% APF 겔 도포군, 그리고 5% NaF 바니쉬 도포군으로 구성하여 실험을 진행하였으며, 불소도포 전후의 양상은 CLSM을 이용하여 평균형광강도로 확인하였다. 불소도포 전 후 평균형광강도는 모든 군에서 유의하게 감소하는 양상을 보였으나, 평균형광강도변화량은 실험군간의 통계적으로 유의한 차이는 발견되지 않았다(p=0.222). 탈회된 표면의 표면미세경도와 평균형광강도 간에는 음의 상관관계를 보였고(p=0.941), 탈회표면의 평균형광강도가 클수록 불소도포 1일 경과 후 평균형광강도 증가하는 양의 상관관계를 보였다(r=0.811, p<0.001). 차 후 구강 내 환경을 반영한 모델이나 실제 in-situ 또는 임상실험을 통해 불소도포제제의 재광화 양상에 대한 연구가 필요할 것이다.

SURFACE CHARACTERISTICS OF ANODIC OXIDIZED TITANIUM ACCORDING TO THE PORE SIZE

  • Ha Heon-Seok;Kim Chang-Whe;Lim Young-Jun;Kim Myung-Joo
    • 대한치과보철학회지
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    • 제44권3호
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    • pp.343-355
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    • 2006
  • Statement of problem. The success of osseointegration can be enhanced with an implant that has improved surface characteristics. Anodic oxidation is one of the surface modifying method to achieve osseointegration. Voltage of anodic oxidation can change surface characteristics and cell activity Purpose. This study was performed to evaluate MG63 cell responses such as affinity, proliferation and to compare surface characteristics of anodic oxidized titanium in various voltage. Material and method. The disks for cell culture were fabricated from grade 3 commercially pure titanium,1 m in thickness and 12 mm in diameter. Surfaces of 4 different roughness were prepared. Group 1 had a machined surface, used as control. Group 2 was anodized under 220 V, group 3 was anodized under 300 V and group 4 was anodized under 320 V. The microtopography of specimens was observed by scanning electron microscope (JSM-840A, JEOL, Japan) and atomic force microscope(Autoprobe CP, Park Scientific Instrument, USA). The surface roughness was measured by confocal laser scanning microscope(Pascal, LSM5, Zeiss, Germany). The crystal structure of the titanium surface was analyzed with x-ray diffractometer(D8 advanced, Broker, Germany). MG63 osteoblast-like cells were cultured on these specimens. The cell morpholgy was observed by field emission electron microscope(Hitachi S-4700, Japan). The cell metabolic and proliferative activity was evaluated by MTT assay Results and conclusion. With in limitations of this in vitro study, the following conclusions were drawn. 1. In anodizing titanium surface, we could see pores which did not show in control group. In higher anodizing voltage, pore size was increased. 2. In anodizing titanium surface, we could see anatase. In higher anodizing voltage, thicker oxide layer increased crystallinity(anatase, anatase and rutile mixed). 3. MG63 cells showed more irregular, polarized and polygonal shape and developed more lamellipodi in anodizing group as voltage increased. 4. The activity of cells in MTT assay increased significantly in group 3 and 4 in comparison with group 1 and 2. However, there was no difference between group 3 and 4 at P<0.05. Proliferation of MG63 cells increased significantly in pore size($3-5.5{\mu}m$) of group 3 and 4 in comparison with in pore size($0.2-1{\mu}m$ ) of group 2.

경락추적을 위한 피내 알시안 블루 주입방법 (Intradermal Alcian-Blue Injection Method to Trace Acupuncture Meridians)

  • 성백경;김민수;오가이;강대인;소광섭
    • 대한약침학회지
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    • 제11권2호
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    • pp.5-12
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    • 2008
  • 목적 : 본 논문에서는 경락을 추적하기 위한 피내(皮內) 알시안 블루(Alcian blue) 염색 방법을 기술한다. 방법 : 1% 알시안 블루 용액을 31게이지 바늘이 달린 0.5mL 인슐린 주사기를 사용하여 경혈 지점에 피내 주사한 후 실체 현미경하에서 수술, 관찰하였다. 면역조직화학적 방법을 사용하여 해당 조직을 레이저 공초점주사현미경으로 관찰하였다. 결과 : 알시안 블루로 염색되고 피하로 이어지는 실 모양 구조물을 관찰하였다. 이 조직 내에서 특징적인 막대모양 핵 배열 및 $1-2{\mu}m$크기의 데옥시리보핵산 입자가 관찰되었다. 또한 경혈 조직 내에서 풍부한 모세혈관총 및 말초신경 말단, 그리고 약 $300{\mu}m$크기의 소체형 구조물을 확인하였다. 결론 : 경혈 지점의 피내에서 발견된 특징적 실 모양 구조물 및 소체형 구조물은 각각 표층 봉한관 및 소체로 판단된다.

Analysis of osteogenic potential on 3mol% yttria-stabilized tetragonal zirconia polycrystals and two different niobium oxide containing zirconia ceramics

  • Hein, Aung Thu;Cho, Young-Dan;Jo, Ye-Hyeon;Kim, Dae-Joon;Han, Jung-Suk
    • The Journal of Advanced Prosthodontics
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    • 제10권2호
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    • pp.147-154
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    • 2018
  • PURPOSE. This study was performed to evaluate the osteogenic potential of 3mol% yttria-stabilized tetragonal zirconia polycrystals (3Y-TZP) and niobium oxide containing Y-TZPs with specific ratios, new (Y,Nb)-TZPs, namely YN4533 and YN4533/Al20 discs. MATERIALS AND METHODS. 3Y-TZP, YN4533 and YN4533/Al20 discs (15 mm diameter and 1 mm thickness) were prepared and their average surface roughness ($R_a$) and surface topography were analyzed using 3-D confocal laser microscope (CLSM) and scanning electron microscope (SEM). Mouse pre-osteoblast MC3T3-E1 cells were seeded onto all zirconia discs and evaluated with regard to cell attachment and morphology by (CLSM), cell proliferation by PicoGreen assay, and cell differentiation by Reverse-Transcription PCR and Quantitative Real-Time PCR, and alkaline phosphatase (Alp) staining. RESULTS. The cellular morphology of MC3T3-E1 pre-osteoblasts was more stretched on a smooth surface than on a rough surface, regardless of the material. Cellular proliferation was higher on smooth surfaces, but there were no significant differences between 3Y-TZP, YN4533, and YN4533/Al20. Osteoblast differentiation patterns on YN4533 and YN4533/Al20 were similar to or slightly higher than seen in 3Y-TZP. Although there were no significant differences in bone marker gene expression (alkaline phosphatase and osteocalcin), Alp staining indicated better osteoblast differentiation on YN4533 and YN4533/Al20 compared to 3Y-TZP. CONCLUSION. Based on these results, niobium oxide containing Y-TZPs have comparable osteogenic potential to 3Y-TZP and are expected to be suitable alternative ceramics dental implant materials to titanium for aesthetically important areas.

Development of Hair Keratin Protein to Accelerate Oral Mucosal Regeneration

  • So-Yeon Kim
    • 치위생과학회지
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    • 제23권4호
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    • pp.369-377
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    • 2023
  • Background: In this study, we investigated the potential use of keratin for oral tissue regeneration. Keratin is well-known for its effectiveness in skin regeneration by promoting keratinization and enhancing the elasticity and activity of fibroblasts. Because of its structural stability, high storability, biocompatibility, and safety in humans, existing research has predominantly focused on its role in skin wound healing. Herein, we propose using keratin proteins as biocompatible materials for dental applications. Methods: To assess the suitability of alpha-keratin protein as a substrate for cell culture, keratin was extracted from human hair via PEGylation. Viabilities of primary human gingival fibroblasts (HGFs) and human oral keratinocytes (HOKs) were assessed. Fluorescence immunostaining and migration assays were conducted using a fluorescence microscope and confocal laser scanning microscope. Wound healing and migration assays were performed using automated software to analyze the experimental readout and gap closure rate. Results: We confirmed the extraction of alpha-keratin and formation of the PEG-g-keratin complex. Treatment of HGFs with keratin protein at a concentration of 5 mg/ml promoted proliferation and maintained cell viability in the test group compared to the control group. HOKs treated with 5 mg/ml keratin exhibited a slight decrease in cell proliferation and activity after 48 hours compared to the untreated group, followed by an increase after 72 hours. Wound healing and migration assays revealed rapid closure of the area covered by HOKs over time following keratin treatment. Additionally, HOKs exhibited changes in cell morphology and increased the expression of the mesenchymal marker vimentin. Conclusion: Our study demonstrated the potential of hair keratin for soft tissue regeneration, with potential future applications in clinical settings for wound healing.

균질화 해석법을 이용한 단열 간극변화에 따른 투수계수 해석 (The homogenization analysis for permeability coefficients by fracture aperture variations)

  • 채병곤
    • 지질공학
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    • 제14권1호
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    • pp.47-60
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    • 2004
  • 이 연구에서는 간극변화에 따른 단열 기하양상을 충분히 고려한 투수계수를 구하고자 균질화 해석법을 이용하여 투수계수를 산정 하였다. 공초점 레이저 스캔 현미경을 이용하여 압력 단계별 간극을 측정한 결과 한개 시료 내에서 각 측정지점별 간극 크기는 모두 다르게 나타나며 단열 면 양쪽이 서로 평행하지 않고 불 평탄한 양상임을 잘 나타낸다. 각 시료별로 압력 단계별 간극양상을 이용하여 단열모델을 각각 작성하고 균질화 해석법을 통해 간극 양상에 따른 투수성 변화를 살펴본 결과, 각 시료별로 산정한 투수계수는 $10^{-1}~10^{-3}cm/sec$의 범위에 분포한다. 시료들은 대체로 압력이 증가함에 따라 일반적으로 투수계수가 감소하는 양상을 나타낸다. 그러나, 시료별로 압력에 따른 투수계수 감소비율이 일정하지 않고 다양한 변화양상을 보인다. 이와 같은 양상은 Chae et al.(2003)의 관찰결과와 잘 일치하며, 이는 간극의 변화가 투수성에 미치는 영향이 크다는 것을 입증한다. 동일 시료 내에서도 3등분한 지점별로 각기 다른 투수계수 값이 계산되었다. 이는 투수계수가 간극의 크기와 분포형태에 따라 민감하게 변화함을 지시하는 것이다. 따라서, 간극 분포가 일정하지 않은 암석 내 단열에서의 투수특성 해석 시에는 정확한 단열 기하양상을 반영하는 것이 매우 중요하다. 따라서, 단열 내 투수계수 산정을 위해서는 단열 기하양상을 충분히 고려할 필요가 있다.