• Title/Summary/Keyword: confocal microscopy

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Analysis of surface characteristics of (Y, Nb)-TZP after finishing and polishing

  • Seong-keun, Yoo;Ye-Hyeon, Jo;In-Sung Luke, Yeo;Hyung-In, Yoon;Jae-Hyun, Lee;Jin-Soo, Ahn;Jung-Suk, Han
    • The Journal of Advanced Prosthodontics
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    • v.14 no.6
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    • pp.335-345
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    • 2022
  • PURPOSE. This in vitro study aimed to evaluate the surface characteristics of a full veneer crown fabricated chairside (CS) from a (Y, Nb)-TZP zirconia block in response to conventional zirconia grinding and polishing. MATERIALS AND METHODS. Zirconia crowns (n = 40) were first prepared and divided into two groups of materials: Labside (LS) and CS, after which each specimen went through a five-step grinding and polishing procedure. Following each surface treatment, surface characteristics were analyzed using confocal laser microscopy (CLSM), average surface roughness (Ra) values were processed from the profile data through Gaussian filtering, and X-ray diffraction pattern analysis was performed to evaluate the monoclinic (M) phase content. Then, a representative specimen was selected for field-emission scanning electron microscopy (FE-SEM), followed by a final analysis of the roughness and X-ray diffraction of the specimens using the independent t-test and repeated measures analysis of variance (RM-ANOVA). RESULTS. In every group, polishing significantly reduced the Ra values (P < .001). There was no significant difference in Ra between the polished state CS and LS. Furthermore, CLSM and FE-SEM investigations revealed that even though grain exposure was visible in CS specimens throughout the as-delivered and ground states, the exposure was reduced after polishing. Moreover, while no phase transformation was visible in the LS, phase transformation was visible in CS after every surface treatment, with the M phase content of the CS group showing a significant reduction after polishing (P < .001). CONCLUSION. Within the limits of this study, clinically acceptable level of surface finishing of (Y, Nb)-TZP can be achieved after conventional zirconia polishing sequence.

Clinical comparison of marginal fit of ceramic inlays between digital and conventional impressions

  • Franklin Guillermo Vargas-Corral;Americo Ernesto Vargas-Corral;Miguel Angel Rodríguez Valverde;Manuel Bravo;Juan Ignacio Rosales Leal
    • The Journal of Advanced Prosthodontics
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    • v.16 no.1
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    • pp.57-65
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    • 2024
  • PURPOSE. The aim of this stuldy was to compare the clinical marginal fit of CAD-CAM inlays obtained from intraoral digital impression or addition silicone impression techniques. MATERIALS AND METHODS. The study included 31 inlays for prosthodontics purposes of 31 patients: 15 based on intraoral digital impressions (DI group); and 16 based on a conventional impression technique (CI group). Inlays included occlusal and a non-occlusal surface. Inlays were milled in ceramic. The inlay-teeth interface was replicated by placing each inlay in its corresponding uncemented clinical preparation and taking interface impressions with silicone material from occlusal and free surfaces. Interface analysis was made using white light confocal microscopy (WLCM) (scanning area: 694 × 510 ㎛2) from the impression samples. The gap size and the inlay overextension were measured from the microscopy topographies. For analytical purposes (i.e., 95-%-confidence intervals calculations and P-value calculations), the procedure REGRESS in SUDAAN was used to account for clustering (i.e., multiple measurements). For p-value calculation, the log transformation of the dependent variables was used to normalize the distributions. RESULTS. Marginal fit values for occlusal and free surfaces were affected by the type of impression. There were no differences between surfaces (occlusal vs. free). Gap obtained for DI group was 164 ± 84 ㎛ and that for CI group was 209 ± 104 ㎛, and there were statistical differences between them (p = .041). Mean overextension values were 60 ± 59 ㎛ for DI group and 67 ± 73 ㎛ for CI group, and there were no differences between then (p = .553). CONCLUSION. Digital impression achieved inlays with higher clinical marginal fit and performed better than the conventional silicone materials.

Red ginseng extract inhibits lipopolysaccharide-induced platelet-leukocyte aggregates in mice

  • Yuan Yee Lee;Sung Dae Kim;Jin-Kyu Park;Won-Jae Lee;Jee Eun Han;Min-Soo Seo;Min-Goo Seo;Seulgi Bae;Dongmi Kwak;Evelyn Saba;Man Hee Rhee
    • Journal of Ginseng Research
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    • v.48 no.4
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    • pp.428-434
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    • 2024
  • Background: Platelet-leukocyte aggregates (PLAs) play important roles in cardiovascular disease and sepsis. Red ginseng extract (RGE) has been well-studied for its antiplatelet and anti-inflammatory activities. However, the potential inhibitory effects of RGE on PLA have not been investigated. Methods: Six-week-old ICR mice were given oral gavage of RGE for 7 days, followed by an intraperitoneal injection of 15 mg/kg of lipopolysaccharide. Mice were euthanized 24 h later, and blood samples were collected for further analysis. Flow cytometry was utilized to sort populations of PLAs and platelet-neutrophil aggregates (PNAs). By using confocal microscopy, PNAs were validated. Morphological changes in platelets and leukocytes were visualized with scanning electron microscopy. Expressions of tissue factor (TF) and platelet factor 4 (PF4) were investigated using enzyme-linked immunosorbent assay. Results: Populations of activated platelets, PLAs and PNAs, were significantly increased with LPS-induction. Treatment with 200 and 400 mg/kg of RGE decreased platelet activation. Moreover, the populations of PLAs and PNAs were reduced. PNAs were visible in the blood of septic mice, and this was attenuated by treatment with 400 mg/kg of RGE. Morphologically, sepsisinduced platelet activation and fibrin formation in the blood. This was reduced with RGE treatment. Sepsis-induced increase in the plasma levels of TF and PF4 was also reduced with RGE treatment. Conclusion: This study shows that RGE is a potential therapeutic that reduces the activation of platelets and targets PLA and PNA formation. Detailed inhibitory mechanisms of RGE should be studied.

The Changes of Occludin in Tight Junction of Blood-Brain Barrier by ROS (치밀이음부 구조단백질인 Occludin에 대한 활성산소종의 영향)

  • Lee, Hee-Sang;Kim, Dae-Jin;Sohn, Dong-Suep;Jeong, Bong-Su;Choi, Hyung-Taek;Sim, Kyu-Min;Lee, Keum-Jeong;Cho, Hye-Jin;Kim, Suk-Joong;Lee, Jong-Chan;Jeong, Yoon-Hee;Kim, Sung-Su;Lee, Won-Bok
    • Applied Microscopy
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    • v.34 no.4
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    • pp.231-239
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    • 2004
  • Cerebral microvessel endothelial cells that form blood-brain barrier (BBB) have tight junction for maintaining brain homeostasis. Occludin, one of tight junction protein, is crucial for BBB function. $H_2O_2$ induced occludin changes and effects in bovine brain BBB endothelial cells were examined in this study. The decrease of transendothelial electrical resistance (TEER) by $H_2O_2$ was due to disruption of occludin localization. Cytotoxicity test revealed that $H_2O_2$ did not cause cell death below 1 mM $H_2O_2$ within 4 hr. $H_2O_2$ caused intermittent disruption and loss of occludin at tight junctions and occludin disappeared with dose dependent manner from tight junction in confocal laser microscopy. But Western blot revealed that the total amounts of occludin increased by $H_2O_2$ administration. Transmission electron microscopy revealed that the ultrastructure of tight junction was not changed by $H_2O_2$. These data suggest that functional disruption of BBB by $H_2O_2$ was due to the localized loss of occludin in tight junction, but the expression of occludin increased in order to compensate the disrupted function in BBB.

Mitochondrial oxidative phosphorylation complexes exist in the sarcolemma of skeletal muscle

  • Lee, Hyun;Kim, Seung-Hyeob;Lee, Jae-Seon;Yang, Yun-Hee;Nam, Jwa-Min;Kim, Bong-Woo;Ko, Young-Gyu
    • BMB Reports
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    • v.49 no.2
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    • pp.116-121
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    • 2016
  • Although proteomic analyses have revealed the presence of mitochondrial oxidative phosphorylation (OXPHOS) proteins in the plasma membrane, there have been no in-depth evaluations of the presence or function of OXPHOS I-V in the plasma membrane. Here, we demonstrate the in situ localization of OXPHOS I-V complexes to the sarcolemma of skeletal muscle by immunofluorescence and immunohistochemistry. A portion of the OXPHOS I-V complex proteins was not co-stained with MitoTracker but co-localized with caveolin-3 in the sarcolemma of mouse gastrocnemius. Mitochondrial matrix-facing OXPHOS complex subunits were ectopically expressed in the sarcolemma of the non-permeabilized muscle fibers and C2C12 myotubes. The sarcolemmal localization of cytochrome c was also observed from mouse gastrocnemius muscles and C2C12 myotubes, as determined by confocal and total internal resonance fluorescence (TIRF) microscopy. Based on these data, we conclude that a portion of OXPHOS complexes is localized in the sarcolemma of skeletal muscle and may have non-canonical functions.

Fast temporal detection of intracellular hydrogen peroxide by HyPer

  • Yang, Yu-Mi;Lee, Sung Jun;Shin, Dong Min
    • International Journal of Oral Biology
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    • v.38 no.4
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    • pp.169-173
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    • 2013
  • HyPer is the genetically encoded biosensor of intracellular hydrogen peroxide ($H_2O_2$), the most stable of the reactive oxygen species (ROS) generated by living cells. HyPer has a high sensitivity and specificity for detecting intracellular $H_2O_2$ by confocal laser microscopy. However, it was not known whether high speed ratiometric imaging of $H_2O_2$ by HyPer is possible. We thus investigated the sensitivity of HyPer in detecting changes to the intracellular $H_2O_2$ levels in HEK293 and PC12 cells using a microfluorometer imaging system. Increase in the HyPer ratio were clearly evident on stimulations of more than $100{\mu}M$ $H_2O_2$ and fast changes in the HyPer ratio were observed on ratiometric fluorescent images after $H_2O_2$ treatment. These results suggest that HyPer is a potent biosensor of the fast temporal production of intracellular $H_2O_2$.

Swiprosin-1 Regulates Cytokine Expression of Human Mast Cell Line HMC-1 through Actin Remodeling

  • Ramesh, T.P.;Kim, Young-Dae;Kwon, Min-Sung;Jun, Chang-Duk;Kim, Sang-Wook
    • IMMUNE NETWORK
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    • v.9 no.6
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    • pp.274-284
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    • 2009
  • Background: Swiprosin-1 was identified in human CD8+ lymphocytes, mature B cells and non-lymphonoid tissue. We have recently reported that swiprosin-1 is expressed in mast cells and up-regulated in both in vitro and in vivo. Methods: The expression of cytokines and swiprosin-1 were determined by by real time PCR and conventional PCR. Pharmacological inhibitors were treated to investigate potential mechanism of swiprosin-1 in mast cell activation. Actin content was evaluated by confocal microscopy and flow cytometry. Results: The swiprosin-1 augmented PMA/A23187-induced expression of cytokines and release of histamine. However, knock-down of swiprosin-1 showed only a modest effect on PMA/A23187-induced cytokine expression, suggesting that swiprosin-1 has gain-of-function characteristics. Swiprosin-1 was found in microvilli-like membrane protrusions and highly co-localized with F-actin. Importantly, either disruption of actin by cytochalasin B or inhibition of PI3 kinase, an enzyme involved in actin remodeling, by wortmannin blocked cytokine expression only in swiprosin-1-overexpressing cells. Conclusion: These results suggest that swiprosin-1 modulates mast cell activation potentially through actin regulation.

The study on the influence of surface cleanness and water soluble salt on corrosion protection of epoxy resin coated carbon steel

  • Shon, MinYoung
    • Corrosion Science and Technology
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    • v.13 no.5
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    • pp.163-169
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    • 2014
  • The corrosion resistance of epoxy-coated carbon steel was evaluated. The carbon steel surface was subjected to different treatment methods such as steel grit blasting and power tool treatment as well as contamination of water soluble salt. To study the effect of the surface treatments and contamination, the topology of the treated surface was observed by confocal microscopy and a pull-off adhesion test was conducted. The corrosion resistance of the epoxy-coated carbon steel was further examined by electrochemical impedance spectroscopy (EIS) combined with immersion test of 3.5 wt% of NaCl solution. Consequently, the surface contamination by sodium chloride with $16mg/m^2$, $48mg/m^2$ and $96mg/m^2$ didn't affect the adhesion strength for current epoxy coated carbon steel and blister and rust were not observed on the surface of epoxy coating contaminated by various concentration of sodium chloride after 20 weeks of immersion in 3.5 wt% NaCl aqueous solutions. In addition, the results of EIS test showed that the epoxy-coated carbon steel treated with steel grit blasting and power tool showed similar corrosion protection performance and surface cleanness such as Sa 3 and Sa 2.5 didn't affect the corrosion protectiveness of epoxy coated carbon steel.

Effect of Probiotic Clostridium butyricum NCTC 7423 Supernatant on Biofilm Formation and Gene Expression of Bacteroides fragilis

  • Shi, Da-Seul;Rhee, Ki-Jong;Eom, Yong-Bin
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.368-377
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    • 2020
  • Enterotoxigenic Bacteroides fragilis (ETBF) is the main pathogen causing severe inflammatory diseases and colorectal cancer. Its biofilm plays a key role in the development of colorectal cancer. The objective of this study was to determine the antagonistic effects of cell-free supernatants (CFS) derived from Clostridium butyricum against the growth and biofilm of ETBF. Our data showed that C. butyricum CFS inhibited the growth of B. fragilis in planktonic culture. In addition, C. butyricum CFS exhibited an antibiofilm effect by inhibiting biofilm development, disassembling preformed biofilms and reducing the metabolic activity of cells in biofilms. Using confocal laser scanning microscopy, we found that C. butyricum CFS significantly suppressed the proteins and extracellular nucleic acids among the basic biofilm components. Furthermore, C. butyricum CFS significantly downregulated the expression of virulence- and efflux pump-related genes including ompA and bmeB3 in B. fragilis. Our findings suggest that C. butyricum can be used as biotherapeutic agent by inhibiting the growth and biofilm of ETBF.

SIMS Protein imaging with nanoparticle tagged antibody for simultaneous omic imaging

  • Lee, Seon-Yeong;Mun, Dae-Won
    • Proceedings of the Korean Vacuum Society Conference
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    • 2015.08a
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    • pp.230.1-230.1
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    • 2015
  • One of the major problems of biological ToF-SIMS imaging is the lack of protein and peptide imaging. Most of biological story telling is mianly based on proteins. The biological implication of lipid ToF-SIMS imaging would be much higher if protein imaging is provided together. Utilizing high secondary ion yields of metals, proteins can be ToF-SIMS imaged with nanoparticle tagged proteins. Nanoparticles such as Fe3O4, SiO2, PbS were used for imaing NeuN, MCH, Orexin A, ${\alpha}$ synucline, TH(Tryosine Hydroxylase) in mouse tissues with a spatial resolution of ${\sim}2{\mu}m$ using a TOF-SIMS. Lipids and neurotransmitters images obtained simultaneously with protein images were overlayed for more deeper understanding of neurobiology, which is not allowed by any other bioimaging technqiues. The protein images from TOF-SIMS were compared with confocal fluorescence microscopy and NanoSIMS images. A new sample preparation method for imaging single cell membranes in a tissue using the vibrotome technique to prepare a tissue slice without any fixation and freeze drying will be also presented briefly for Hippocampus and Hypothalamus tissues.

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