• 제목/요약/키워드: laser confocal system

검색결과 86건 처리시간 0.026초

Evaluation of Volumetric Texture Features for Computerized Cell Nuclei Grading

  • Kim, Tae-Yun;Choi, Hyun-Ju;Choi, Heung-Kook
    • 한국멀티미디어학회논문지
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    • 제11권12호
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    • pp.1635-1648
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    • 2008
  • The extraction of important features in cancer cell image analysis is a key process in grading renal cell carcinoma. In this study, we applied three-dimensional (3D) texture feature extraction methods to cell nuclei images and evaluated the validity of them for computerized cell nuclei grading. Individual images of 2,423 cell nuclei were extracted from 80 renal cell carcinomas (RCCs) using confocal laser scanning microscopy (CLSM). First, we applied the 3D texture mapping method to render the volume of entire tissue sections. Then, we determined the chromatin texture quantitatively by calculating 3D gray-level co-occurrence matrices (3D GLCM) and 3D run length matrices (3D GLRLM). Finally, to demonstrate the suitability of 3D texture features for grading, we performed a discriminant analysis. In addition, we conducted a principal component analysis to obtain optimized texture features. Automatic grading of cell nuclei using 3D texture features had an accuracy of 78.30%. Combining 3D textural and 3D morphological features improved the accuracy to 82.19%. As a comparative study, we also performed a stepwise feature selection. Using the 4 optimized features, we could obtain more improved accuracy of 84.32%. Three dimensional texture features have potential for use as fundamental elements in developing a new nuclear grading system with accurate diagnosis and predicting prognosis.

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Development and evaluation of probiotic delivery systems using the rennet-induced gelation of milk proteins

  • Ha, Ho-Kyung;Hong, Ji-Young;Ayu, Istifiani Lola;Lee, Mee-Ryung;Lee, Won-Jae
    • Journal of Animal Science and Technology
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    • 제63권5호
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    • pp.1182-1193
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    • 2021
  • The aims of this study were to develop a milk protein-based probiotic delivery system using a modified rennet-induced gelation method and to determine how the skim milk powder concentration level and pH, which can affect the rennet-induced intra- and inter-molecular association of milk proteins, affect the physicochemical properties of the probiotic delivery systems, such as the particle size, size distribution, encapsulation efficiency, and viability of probiotics in simulated gastrointestinal tract. To prepare a milk protein-based delivery system, skim milk powder was used as a source of milk proteins with various concentration levels from 3 to 10% (w/w) and rennet was added to skim milk solutions followed by adjustment of pH from 5.4 or 6.2. Lactobacillus rhamnosus GG was used as a probiotic culture. In confocal laser scanning microscopic images, globular particles with a size ranging from 10 ㎛ to 20 ㎛ were observed, indicating that milk protein-based probiotic delivery systems were successfully created. When the skim milk powder concentration was increased from 3 to 10% (w/w), the size of the delivery system was significantly (p < 0.05) increased from 27.5 to 44.4 ㎛, while a significant (p < 0.05) increase in size from 26.3 to 34.5 ㎛ was observed as the pH was increased from 5.4 to 6.4. An increase in skim milk powder concentration level and a decrease in pH led to a significant (p < 0.05) increase in the encapsulation efficiency of probiotics. The viability of probiotics in a simulated stomach condition was increased when probiotics were encapsulated in milk protein-based delivery systems. An increase in the skim milk powder concentration and a decrease in pH resulted in an increase in the viability of probiotics in simulated stomach conditions. It was concluded that the protein content by modulating skim milk powder concentration level and pH were the key manufacturing variables affecting the physicochemical properties of milk protein-based probiotic delivery systems.

느타리버섯에서의 Pseudomonas tolaasii오염 연구 (Study on the Contamination of Pseudomonas tolaasii in Oyster Mushroom)

  • 이혜영;장금일;김광엽
    • 한국식품위생안전성학회지
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    • 제16권3호
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    • pp.232-240
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    • 2001
  • 시장에서 수집한 갈변병에 감염된 느타리버섯으로부터 125 균주를 분리하였고 그 중 45 균주는 병원성 균주로 조사되었다. WLFO 6 strains, WLFO 6 strains으로 나타났고, WLRO의 몇 균주는 병원성 검정 실험에서 약한 병원성을 나타내기도 했다. WLFO 6균주는 모두 느타리버섯의 갈변병 주원인균으로 알려진 P. tolaasii로 미생물 신속 동성을 (MIDI, gas chromatograph-microbial identification system)에 의해 동정되었고 WLRO는 P. gingeri, P. fluorescens biotype A and type C. 로 동정되었다. 그 외의 선발된 Pseudomonas spp.는 P. gingeri, P. agarici, P. fluorescens biotype B, P. chloroaphis, non-pathogenic P. tolaasii, P. putida biotype A, B 등으로 동정이 되었다. 분리된 병원성 세균의 세포배양 여과액으로 버섯에서의 갈변과 조직 함몰을 실험하였다. 병원성 검정에서 약한 병원성을 보인 분리균들은 갈변 또는 조직함몰이 나타나지 않았으나 강한 병원성을 나타냈던 균들은 두 현상이 모두 나타났다. P. tolaasii에 의해서 생성되는 세포외독소인 tolaasin의 활성을 조사하기 위하여 600 nm에서 용혈 활성을 측정하였다. P. tolaasii로 동정된 6 균주는 0.8∼0.9, 약한 병원성의 WLROs는 0.9∼1.0 그리고 Pseudomonas spp.는 10∼1.2로 나타났다. 공초점 현미경 기술을 이용하여 신선한 버섯에서의 조직을 optical sectioning image와 vertical sectioning image로 관찰하였고 또한 P. toiaasii에 의하여 오염된 조직부위의 영상을 회득하였다.

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Autophagy-related protein LC3 and Beclin-1 in the first trimester of pregnancy

  • Chifenti, Barbara;Locci, Maria Teresa;Lazzeri, Gloria;Guagnozzi, Mariangela;Dinucci, Dino;Chiellini, Federica;Filice, Maria Elena;Salerno, Maria Giovanna;Battini, Lorella
    • Clinical and Experimental Reproductive Medicine
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    • 제40권1호
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    • pp.33-37
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    • 2013
  • Autophagy is a degradation process that acts in response to environmental stressors. Recently, autophagy has been detected in normal term, preeclamptic and intrauterine growth-restricted placentas. The object of this work was to investigate the presence of autophagy in first trimester voluntary interruption of pregnancy placental villi by the expression of autophagy-related proteins, light chain 3 (LC3), and Beclin-1. In first trimester placental villi laser scanning confocal microscopy (LSCM) analysis revealed LC3 and Beclin-1 immunoreactivity prevalently located in villous cytotrophoblasts. Using LSCM, LC3, and Beclin-1 were localized to the cytoplasm of the trophoblast layer in human full-term placentas. Beclin-1 expression and LC3 activation were confirmed by western blotting. These data emphasize that autophagy activation is different among cytotrophoblasts and syncytiotrophoblasts depending on the gestational age and thus we speculate that autophagy might play a prosurvival role throughout human pregnancy.

Encapsulation of Lactobacillus rhamnosus GG Using Milk Protein-Based Delivery Systems: Effects of Reaction Temperature and Holding Time on Their Physicochemical and Functional Properties

  • Ayu, Istifiani Lola;Ha, Ho-Kyung;Yang, Dong-Hun;Lee, Won-Jae;Lee, Mee-Ryung
    • 한국축산식품학회지
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    • 제41권5호
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    • pp.894-904
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    • 2021
  • Microencapsulation is a protective process for materials that are sensitive to harsh conditions encounted during food manufacture and storage. The objectives of this research were to manufacture a milk protein-based delivery system (MPDS) containing Lactobacillus rhamnosus GG (LGG) using skim milk powder and to investigate the effects of manufacturing variables, such as reaction temerpature and holding time, on the physiccohemical properties of MPDS and viability of LGG under dairy food processing and storage conditions. MPDS was prepared using chymosin at varing reaction temperatures from 25℃ to 40℃ for 10 min and holding times from 5 to 30 min at 25℃. The morphological and physicochemical properties of MPDS were evaluated using a confocal laser scanning microscope and a particle size analyzer, respectively. The number of viable cells were determined using the standard plate method. Spherical-shaped MPDS particles were successfully manufactured. The particle size of MPDS was increased with a decrease in reaction temperature and an increase in holding time. As reaction temperature and holding time were increased, the encapsulation efficiency of LGG in MPDS was increased. During pasteurization, the use of MPDS resulted in an increase in the LGG viability. The encapsulation of LGG in MPDS led to an increase in the viability of LGG in simulated gastric fluid. In addition, the LGG viability was enhanced with an increase in reaction temperature and holding time. In conclusions, the encapsulation of LGG in MPDS could be an effective way of improving the viability of LGG during pasturization process in various foods.

지대치삭제의정확도가레이져신터링기술로제작된Co-Cr 코핑의변연적합도에미치는영향 (Influence of the accuracy of abutment tooth preparation on the marginal adaptation of Co-Cr alloy copings fabricated with a selective laser sintering technology)

  • 김서랑;곽재영;허성주;김성균;김명주
    • 대한치과보철학회지
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    • 제53권4호
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    • pp.337-344
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    • 2015
  • 목적: 본 연구의 목적은 지대치 삭제의 정확도와 metal coping의 변연적합도와의 상관관계를 알아보는 데 있다. 이를 통해, 세 가지 다른 제작 방식 (주조, milling, 기법)이 이 상관관계에 미치는 영향을 비교해 보고자 한다. 재료 및 방법: 두 개의 master medel을 서로 다른 방식으로 제작하였다; 첫 번째 모델은 치과의사에 의해 deep chamfer margin을 가지도록 지대치 삭제된 것이고, 두 번째 모델은 3-D designing software program을 이용하여 동일한 삭제원칙에 따라 제작되었다. 각각의 모델에 대하여 세 가지 제작 방식으로 코발트-크롬 코핑을 12개씩 제작하여, 총 72개의 시편을 얻었다. 각 시편을 master model상에 적합시키고 공초점 레이저 주사 현미경으로 변연적합도를 측정하였다. 결과: SLS system (P=.0231)과 주조방식(P<.0001)에서는 computer designed model이 hand prepared model에 비하여 유의하게 우수한 변연적합도를 보였다. 그러나 milling group에서는 두 모델 간에 유의한 차이를 나타내지 않았다(P=.9962). 결론: 본 연구에 한하여, 지대치 삭제의 정확도가 금속 코핑의 변연적합도에 미치는 영향은 그 제작 방식에 따라 달랐다. 제작 방식에 따른 변연적 합도는 SLS system이 가장 우수하였고 지대치 삭제의 정확도에 의해 영향을 받았다. 한편, milling 방식은 세 가지 방식 중 가장 큰 변연 격차를 나타내었으며 지대치 삭제의 정확도에 영향을 받지 않았다.

Cellular Uptake and Cytotoxicity of β-Lactoglobulin Nanoparticles: The Effects of Particle Size and Surface Charge

  • Ha, Ho-Kyung;Kim, Jin Wook;Lee, Mee-Ryung;Jun, Woojin;Lee, Won-Jae
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권3호
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    • pp.420-427
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    • 2015
  • It is necessary to understand the cellular uptake and cytotoxicity of food-grade delivery systems, such as ${\beta}$-lactoglobulin (${\beta}$-lg) nanoparticles, for the application of bioactive compounds to functional foods. The objectives of this study were to investigate the relationships between the physicochemical properties of ${\beta}$-lg nanoparticles, such as particle size and zeta-potential value, and their cellular uptakes and cytotoxicity in Caco-2 cells. Physicochemical properties of ${\beta}$-lg nanoparticles were evaluated using particle size analyzer. Flow cytometry and confocal laser scanning microscopy were used to investigate cellular uptake and cytotoxicity of ${\beta}$-lg nanoparticles. The ${\beta}$-lg nanoparticles with various particle sizes (98 to 192 nm) and zeta-potential values (-14.8 to -17.6 mV) were successfully formed. A decrease in heating temperature from $70^{\circ}C$ to $60^{\circ}C$ resulted in a decrease in the particle size and an increase in the zeta-potential value of ${\beta}$-lg nanoparticles. Non-cytotoxicity was observed in Caco-2 cells treated with ${\beta}$-lg nanoparticles. There was an increase in cellular uptake of ${\beta}$-lg nanoparticles with a decrease in particle size and an increase in zeta-potential value. Cellular uptake ${\beta}$-lg nanoparticles was negatively correlated with particle size and positively correlated with zeta-potential value. Therefore, these results suggest that the particle size and zeta-potential value of ${\beta}$-lg nanoparticles play an important role in the cellular uptake. The ${\beta}$-lg nanoparticles can be used as a delivery system in foods due to its high cellular uptake and non-cytotoxicity.

Rapid and Accurate Detection of Bacillus anthracis Spores Using Peptide-Quantum Dot Conjugates

  • Park, Tae-Jung;Park, Jong-Pil;Seo, Gwi-Moon;Chai, Young-Gyu;Lee, Sang-Yup
    • Journal of Microbiology and Biotechnology
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    • 제16권11호
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    • pp.1713-1719
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    • 2006
  • A method for the simple, rapid, specific, and accurate detection of Bacillus anthracis spores was developed by employing specific capture peptides conjugated with fluorescent quantum dots (QDs). It was possible to distinguish B. anthracis spores from the spores of B. thuringiensis and B. cereus using these peptide-QD conjugates by flow cytometric and confocal laser scanning microscopic analyses. For more convenient high-throughput detection of B. anthracis spores, spectrofluorometric analysis of spore-peptide-QD conjugates was performed. B. anthracis spores could be detected in less than 1 h using this method. In order to avoid any minor yet false-positive signal caused by the presence of B. thuringiensis spores, the B-Negative peptide, which can only bind to B. thuringiensis, conjugated with another type of QD that fluoresces at different wavelength was also developed. In the presence of mixed B. anthracis and B. thuringiensis spores, the BABA peptide conjugated with QD525 and the B-Negative peptide conjugated with QD585 were able to bind to the former and the latter, specifically and respectively, thus allowing the clear detection of B. anthracis spores against B. thuringiensis spores by using two QD-labeling systems. This capture peptide-conjugated QD system should be useful for the detection of B. anthracis spores.

Monitoring microRNAs Using a Molecular Beacon in CD133+/CD338+ Human Lung Adenocarcinoma-initiating A549 Cells

  • Yao, Quan;Sun, Jian-Guo;Ma, Hu;Zhang, An-Mei;Lin, Sheng;Zhu, Cong-Hui;Zhang, Tao;Chen, Zheng-Tang
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권1호
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    • pp.161-166
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    • 2014
  • Lung cancer is the most common causes of cancer-related deaths worldwide, and a lack of effective methods for early diagnosis has greatly impacted the prognosis and survival rates of the affected patients. Tumor-initiating cells (TICs) are considered to be largely responsible for tumor genesis, resistance to tumor therapy, metastasis, and recurrence. In addition to representing a good potential treatment target, TICs can provide clues for the early diagnosis of cancer. MicroRNA (miRNA) alterations are known to be involved in the initiation and progression of human cancer, and the detection of related miRNAs in TICs is an important strategy for lung cancer early diagnosis. As Hsa-miR-155 (miR-155) can be used as a diagnostic marker for non-small cell lung cancer (NSCLC), a smart molecular beacon of miR-155 was designed to image the expression of miR-155 in NSCLC cases. TICs expressing CD133 and CD338 were obtained from A549 cells by applying an immune magnetic bead isolation system, and miR-155 was detected using laser-scanning confocal microscopy. We found that intracellular miR-155 could be successfully detected using smart miR-155 molecular beacons. Expression was higher in TICs than in A549 cells, indicating that miR-155 may play an important role in regulating bio-behavior of TICs. As a non-invasive approach, molecular beacons could be implemented with molecular imaging to diagnose lung cancer at early stages.

Interaction between the p75 neurotrophin receptor and a novel adaptor protein

  • Lee, Yun-Hee;Yu, Ji-Hee;Cho, Jung-Sun;Park, Han-Jeong;Lee, Seung-Pyo;Paik, Ki-Suk;Chang, Mi-Sook
    • International Journal of Oral Biology
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    • 제33권2호
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    • pp.71-76
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    • 2008
  • The neurotrophin plays an important role in the development, differentiation and survival of the nervous system in vertebrates. It exerts its cellular effects through two different receptors, the Trk receptor tyrosine kinase neurotrophin receptor and the p75 neurotrophin receptor, a member of the tumor necrosis factor receptor superfamily. Trk and p75 neurotrophin receptors utilize specific target proteins to transmit signals into the cell. An ankyrin-rich membrane spanning protein (ARMS) was identified as a new p75 interacting protein and serves as a novel downstream target of p75 neurotrophin receptor. We sought to delineate the interaction between p75 and ARMS by deletion constructs of p75 and green fluorescent protein (GFP)-tagged ARMS. We examined the interaction between these two proteins after overexpressing them in HEK-293 cells. Using both Western blot analysis and immunocytochemistry followed by confocal laser scanning microscopy, we found out that the intracellular domain of the p75 neurotrophin receptor was important for the interaction with ARMS. The results from this study suggest that ARMS may play an important role for mediating the signals from p75 neurotrophin receptor into the cell.