• 제목/요약/키워드: laser scanning confocal microscopy

검색결과 194건 처리시간 0.027초

Defense-Related Responses in Fruit of the Nonhost Chili Pepper against Xanthomonas axonopodis pv. glycines Infection

  • Chang, Sung Pae;Jeon, Yong Ho;Kim, Young Ho
    • The Plant Pathology Journal
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    • 제32권4호
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    • pp.311-320
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    • 2016
  • Xanthomonas axonopodis pv. glycines (Xag) is a necrotrophic bacterial pathogen of the soybean that causes bacterial pustules and is a nonhost pathogen of the chili pepper. In the current study, chili pepper fruit wound inoculated in planta with Xag 8ra formed necrotic lesions on the fruit surface and induced several structural and chemical barriers systemically in the fruit tissue. The initial defense response included programmed cell death of necrotizing and necrotized cells, which was characterized by nuclear DNA cleavage, as detected by TUNEL-confocal laser scanning microscopy (CLSM), and phosphatidylserine exposure on cell walls distal to the infection site, as detected by Annexin V FLUOS-CLSM. These two responses may facilitate cell killing and enhance transportation of cell wall materials used for cell wall thickening, respectively. The cells beneath the necrotic tissue were enlarged and divided to form periclinal cell walls, resulting in extensive formation of several parallel boundary layers at the later stages of infection, accompanying the deposition of wall fortification materials for strengthening structural defenses. These results suggest that nonhost resistance of chili pepper fruit against the nonhost necrotrophic pathogen Xag 8ra is activated systematically from the initial infection until termination of the infection cycle, resulting in complete inhibition of bacterial pathogenesis by utilizing organspecific in situ physiological events governed by the expression of genes in the plant fruit organ.

초지법에 따른 한지의 물성비교 (Comparison of Physical Properties of Hanjis Made by Different Sheet Forming Processes)

  • 최태호;조남석;최인호;정택상
    • 펄프종이기술
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    • 제33권4호
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    • pp.21-27
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    • 2001
  • Korean traditional paper (Hanji) making technology has adopted two kinds of sheet forming processes, which called "Oebal-choji": and "Ssangbal-choji". The sheet forming process of Oebal-choji is an original method developed in Korea. At first, paper stock is dipped onto the mold and flow away in the forward direction. Then, paper stock is scooped again and rhythmically rocked from side to side, this work is repeated several times. Through this operation the fibers intertwine and paper layers are formed. Ssangbal-choji is almost same as the Nagashizuki, which used in Japan. In this method, paper stock is scooped onto the mold and rhythmically rocked backwards and forwards several times, the water drains slowly through the bamboo screen and then sheet is formed. Tamezuki method is used in Japan and China. This is a method in which the mold is dipped into the paper stock once and left to drain. In the Ssangbal-choji and Nagashizuki methods, the most of excess solution is cast out while in the Tamezuki all of it is allowed to drain through the mold. This study was carried out to investigate the physical properties of the Hanjis that were made by Oebal-choji, Ssangbal-choji, Nagashizuki, and Tamezuki sheet forming processes. The results were follows; Physical properties of the Oebal-choji Hanji were better than those of Ssangbal-choji, Nagashizuki, and Tamezuki. Oebal-choji Hanji made little difference of paper strength between MD and CD, but Ssangbal-chjo and Nagashizuki Hanjis made wide difference. And there are no difference of paper strength between MD and CD on the Tamezuki Hanji. On the confocal laser scanning microscopy (CLSM) observation of the Hanjis, Oebal-choji made well crossed fiber orientation than those of other forming processes.r forming processes.

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세륨옥사이드나노입자(Cerium oxide nano particles: CNPs)를 함유한 치면열구전색재의 Streptococcus mutans 부착량 변화 (Changes in the amount of adhesion of Streptococcus mutans to pit and fissure sealant incorporating cerium oxide nano particles(CNPs))

  • 이성숙;박영민;김동애
    • 한국치위생학회지
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    • 제20권4호
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    • pp.535-543
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    • 2020
  • Objectives: The aim of this study was to investigated the surface roughness and change in the amount of adhesion of Streptococcus mutans to the commercial pit and fissure sealant containing cerium oxide nano particles(CNPs). Methods: The CNPs was incorporated into a commercial pit and fissure sealant at 0-4.0 wt%. Disk Specimens (ϕ 10 mm × 2 mm) were prepared by light polymerization the front and back for 40s. Average surface roughness was measured and Streptococcus mutans adhesion was observed under confocal laser scanning microscopy (CLSM) after 24 hour. Data were statistically analyzed by one-way ANOVA and Tukey HSDa post-hoc test. Results: Difference of the surface roughness(Ra) between groups was not statistically significant in both non CNPs group and CNPs group(p>0.05). In CNPs group, the amount of S. mutans adhesion was significantly different between control group and decreased in order of CNPs 4.0, CNPs 0.5, CNPs 1.0 and CNPs 2.0(p<0.05). Conclusions: Within the limitation of this study, these aspects of oral bacteria performances suggest potential usefulness of the CNPs incorporation, especially CNPs 1% and 2%, in pit and fissure sealant for inducing antibacterial effect.

Effects of Reactive Oxygen Species and Nitrogen Species on the Excitability of Spinal Substantia Gelatinosa Neurons

  • Park, Joo Young;Park, Areum;Chun, Sang Woo
    • International Journal of Oral Biology
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    • 제41권3호
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    • pp.141-147
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    • 2016
  • Reactive oxygen species (ROS) and nitrogen species (RNS) are both important signaling molecules involved in pain transmission in the dorsal horn of the spinal cord. Xanthine oxidase (XO) is a well-known enzyme for the generation of superoxide anions ($O_2^{\bullet-}$), while S-nitroso-N-acetyl-DL-penicillamine (SNAP) is a representative nitric oxide (NO) donor. In this study, we used patch clamp recording in spinal slices of rats to investigate the effects of $O_2^{\bullet-}$ and NO on the excitability of substantia gelatinosa (SG) neurons. We also used confocal scanning laser microscopy to measure XO- and SNAP-induced ROS and RNS production in live slices. We observed that the ROS level increased during the perfusion of xanthine and xanthine oxidase (X/XO) compound and SNAP after the loading of 2',7'-dichlorofluorescin diacetate ($H_2DCF-DA$), which is an indicator of intracellular ROS and RNS. Application of ROS donors such as X/XO, ${\beta}-nicotinamide$ adenine dinucleotide phosphate (NADPH), and 3-morpholinosydnomimine (SIN-1) induced a membrane depolarization and inward currents. SNAP, an RNS donor, also induced membrane depolarization and inward currents. X/XO-induced inward currents were significantly decreased by pretreatment with phenyl N-tert-butylnitrone (PBN; nonspecific ROS and RNS scavenger) and manganese(III) tetrakis(4-benzoic acid) porphyrin (MnTBAP; superoxide dismutase mimetics). Nitro-L-arginine methyl ester (NAME; NO scavenger) also slightly decreased X/XO-induced inward currents, suggesting that X/XO-induced responses can be involved in the generation of peroxynitrite ($ONOO^-$). Our data suggest that elevated ROS, especially $O_2^{\bullet-}$, NO and $ONOO^-$, in the spinal cord can increase the excitability of the SG neurons related to pain transmission.

Investigating the Morphology and Kinetics of Three-Dimensional Neuronal Networks on Electro-Spun Microstructured Scaffolds

  • Kim, Dongyoon;Kim, Seong-Min;Kang, Donghee;Baek, Goeun;Yoon, Myung-Han
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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    • pp.277.2-277.2
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    • 2013
  • Petri dishes and glass slides have been widely used as general substrates for in vitro mammalian cell cultures due to their culture viability, optical transparency, experimental convenience, and relatively low cost. Despite the aforementioned benefit, however, the flat two-dimensional substrates exhibit limited capability in terms of realistically mimicking cellular polarization, intercellular interaction, and differentiation in the non-physiological culture environment. Here, we report a protocol of culturing embryonic rat hippocampal neurons on the electro-spun polymeric network and the results from examination of neuronal cell behavior and network formation on this culture platform. A combinatorial method of laser-scanning confocal fluorescence microscopy and live-cell imaging technique was employed to track axonal outgrowth and synaptic connectivity of the neuronal cells deposited on this model culture environment. The present microfiber-based scaffold supports the prolonged viability of three-dimensionally-formed neuronal networks and their microscopic geometric parameters (i.e., microfiber diameter) strongly influence the axonal outgrowth and synaptic connection pattern. These results implies that electro-spun fiber scaffolds with fine control over surface chemistry and nano/microscopic geometry may be used as an economic and general platform for three-dimensional mammalian culture systems, particularly, neuronal lineage and other network forming cell lines.

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Ginsenosides attenuate bioenergetics and morphology of mitochondria in cultured PC12 cells under the insult of amyloid beta-peptide

  • Kwan, Kenneth Kin Leung;Yun, Huang;Dong, Tina Ting Xia;Tsim, Karl Wah Keung
    • Journal of Ginseng Research
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    • 제45권4호
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    • pp.473-481
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    • 2021
  • Background: Mitochondrial dysfunction is one of the significant reasons for Alzheimer's disease (AD). Ginsenosides, natural molecules extracted from Panax ginseng, have been demonstrated to exert essential neuroprotective functions, which can ascribe to its anti-oxidative effect, enhancing central metabolism and improving mitochondrial function. However, a comprehensive analysis of cellular mitochondrial bioenergetics after ginsenoside treatment under Aβ-oxidative stress is missing. Methods: The antioxidant activities of ginsenoside Rb1, Rd, Re, Rg1 were compared by measuring the cell survival and reactive oxygen species (ROS) formation. Next, the protective effects of ginsenosides of mitochondrial bioenergetics were examined by measuring oxygen consumption rate (OCR) in PC12 cells under Aβ-oxidative stress with an extracellular flux analyzer. Meanwhile, mitochondrial membrane potential (MMP) and mitochondrial dynamics were evaluated by confocal laser scanning microscopy. Results: Ginsenoside Rg1 possessed the strongest anti-oxidative property, and which therefore provided the best protective function to PC12 cells under the Aβ oxidative stress by increasing ATP production to 3 folds, spare capacity to 2 folds, maximal respiration to 2 folds and non-mitochondrial respiration to 1.5 folds, as compared to Aβ cell model. Furthermore, ginsenoside Rg1 enhanced MMP and mitochondrial interconnectivity, and simultaneously reduced mitochondrial circularity. Conclusion: In the present study, these results demonstrated that ginsenoside Rg1 could be the best natural compound, as compared with other ginsenosides, by modulating the OCR of cultured PC12 cells during oxidative phosphorylation, in regulating MMP and in improving mitochondria dynamics under Aβ-induced oxidative stress.

Hepatitis B Virus DNA Polymerase Displays an Anti-Apoptotic Effect by Interacting with Elongation Factor-1 Alpha-2 in Hepatoma Cells

  • Niu, Xianli;Nong, Shirong;Gong, Junyuan;Zhang, Xin;Tang, Hui;Zhou, Tianhong;Li, Wei
    • Journal of Microbiology and Biotechnology
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    • 제31권1호
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    • pp.16-24
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    • 2021
  • Hepatitis B virus (HBV) genome P-encoded protein HBV DNA polymerase (Pol) has long been known as a reverse transcriptase during HBV replication. In this study, we investigated the impact of HBV Pol on host cellular processes, mainly apoptosis, and the underlying mechanisms. We showed a marked reduction in apoptotic rates in the HBV Pol-expressed HepG2 cells compared to controls. Moreover, a series of assays, i.e., yeast two-hybrid, GST pull-down, co-immunoprecipitation, and confocal laser scanning microscopy, identified the host factor eEF1A2 to be associated with HBV Pol. Furthermore, knockdown of eEF1A2 gene by siRNA abrogated the HBV Pol-mediated anti-apoptotic effect with apoptosis induced by endoplasmatic reticulum (ER) stress-inducer thapsigargin (TG), thus suggesting that the host factor eEF1A2 is essential for HBV Pol's anti-apoptosis properties. Our findings have revealed a novel role for HBV Pol in its modulation of apoptosis through integrating with eEF1A2.

Tuberostemonine에 의한 Staphylococcus aureus의 생물막 억제 효과 (Inhibitory effects of tuberostemonine on Staphylococcus aureus biofilm)

  • 염수진;김승민;권준혁;정희곤
    • 한국식품과학회지
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    • 제54권2호
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    • pp.241-246
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    • 2022
  • 본 연구는 S. aureus에 대한 tuberostemonine의 항균 및 항생물막 효과에 대하여 확인하였다. S. aureus에 대한 tuberostemonine의 생장 저해 효과가 없음을 확인하였으나, crystal violet 염색법과 CLSM 이미지 측정을 통해 tuberostemonine이 유의한 S. aureus 항생물막 효과 가지는 것을 알 수 있었다. S. aureus의 생물막 형성과 분해 관련 유전자인 icaA와 agrA의 발현은 tuberostemonine를 처리하였을 때 유의미하게 각각 감소 또는 증가하는 것으로 나타났다. 따라서 본 연구에서 생물막 형성 저해 및 분해 효과가 확인된 천연화합물인 tuberostemonine은 S. aureus의 내성 발생 위험이 적은 새로운 항생물막제제로서 사용가능 할 것으로 사료된다.

인공 우식 유발성 biofilm 구성성분의 시간 흐름에 따른 변화 (Changes in the composition of artificial cariogenic biofilms over time)

  • 오철;판딧 싼토스;전재규
    • Journal of Korean Academy of Oral Health
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    • 제43권3호
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    • pp.118-123
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    • 2019
  • Objectives: The purpose of this study was to investigate changes in the composition of artificial cariogenic biofilms using a Streptococcus mutans biofilm model over a period of time. Methods: We analyzed the dry weight, colony forming unit (CFU) number, extracellular polysaccharide (EPS) biovolume, and acid production rate of S. mutans biofilms formed on saliva-coated hydroxyapatite discs after 26 h, 50 h, 74 h, 98 h, 171 h, and 195 h. In addition, we performed a laser scanning confocal fluorescence microscopy to determine the bacterial volume, EPS biovolume, and biofilm thickness. We calculated the biofilm density using dry weight and EPS biovolume. Results: Over a period of time, there was no change in the CFU number and acid production rate of S. mutans biofilms, but there was an increase in the dry weight and EPS biovolume of S. mutans biofilms. The bacterial volume, EPS biovolume, and biofilm thickness only increased in the 50-h-old biofilm; however, no change was observed in 50-195-h-old biofilms. In addition, an increase in the biofilm density was observed over time. Conclusions: These results suggest that the acid production ability of cariogenic biofilms does not change, but the biofilm density increases over time. However, due to scientific information, further research needs to be conducted in the field of dentistry to get further insights on the progression of cariogenic biofilms over time.

수크로스 에스터와 세라마이드를 혼합 겔레이터로 사용한 올레오겔과 올레오겔 에멀젼의 특성 분석 (Characterization of Oleogels and Oleogel Emulsions Made with Sucrose Ester and Ceramide as Mixed Gelators)

  • 이다연;진병석
    • 공업화학
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    • 제34권5호
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    • pp.501-506
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    • 2023
  • 수크로스 에스터와 세라마이드를 혼합하여 겔레이터로 사용한 해바라기 오일의 올레오겔과 올레오겔 에멀전을 제조하였다. 올레오겔과 올레오겔 에멀젼 제형 내에서 겔레이터의 결정 구조는 편광현미경으로 관찰하고 물의 분산형태는 공초점 현미경으로 확인하였다. DSC 열차트 분석을 통해, 수크로스 에스터와 세라마이드를 혼합하면 세라마이드의 결정구조는 사라지고 제형에 물이 첨가되면 혼합된 겔레이터의 결정성이 더욱 상승함을 확인하였다. 제형에서의 수크로스 에스터, 세라마이드, 물의 비율에 따른 점도와 점탄성 등의 유변학적 특성의 변화를 살펴보았다. 세라마이드와 물의 함량이 증가할수록 점도 및 저장탄성률, 손실 탄성률 등이 모두 증가하고, 또한 제형의 안정성도 높아지는 경향을 보였다.