• Korean Journal of Clinical Laboratory Science
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• v.52 no.1
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• pp.1-17
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• 2020

Three-dimensional microscopic approaches in histopathology display multiplex properties that present puzzling questions for specimens as related to their comprehensive volumetric information. This information includes spatial distribution of molecules, three-dimensional co-localization, structural formation and whole data set that cannot be determined by two-dimensional section slides due to the inevitable loss of spatial information. Advancement of optical instruments such as two-photon microscopy and high performance objectives with motorized correction collars have narrowed the gap between optical theories and the actual reality of deep tissue imaging. However, the benefits gained by a prolonged working distance, two-photon laser and optimized beam alignment are inevitably diminished because of the light scattering phenomenon that is deeply related to the refractive index mismatch between each cellular component and the surrounding medium. From the first approaches with simple crude refractive index matching techniques to the recent cutting-edge integrated tissue clearing methods, an achievement of transparency without morphological denaturation and eradication of natural and fixation-induced nonspecific autofluorescence out of real signal are key factors to determine the perfection of tissue clearing and the immunofluorescent staining for high contrast images. When performing integrated laboratory workflow of tissue for processing frozen and formalin-fixed tissues, clear lipid-exchanged acrylamide-hybridized rigid imaging/immunostaining/in situ hybridization-compatible tissue hydrogel (CLARITY), an equipment-based tissue clearing method, is compatible with routine procedures in a histopathology laboratory.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.7
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• pp.857-867
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• 2017

This study was performed to encapsulate low molecular weight marine collagen and ${\gamma}$-aminobutyric acid (GABA)-producing lactic acid bacteria to inhibit degradation and improve survival rate during exposure to adverse conditions of the gastro-intestinal tract. Calcium-alginate method was used for the manufacture of a double-layered coated capsule. The inner core material was composed of collagen and lactic acid bacteria, and the coating materials were alginate and chitosan. The sizes and shapes of the double-coated capsule were affected mainly by centrifuge speed and pH. Manufactured capsules were observed with a scanning electron microscope and by confocal laser scanning microscopy to confirm the micromorphological changes of capsules and bacterial cells. As a result, double-layered coated capsules were not degraded at pH 1.2, whereas degradation occurred at pH 7.4. In addition, GABA and collagen were maintained in stable state at pH 1.2. Therefore, double-layered coated capsules developed in this study would not be degraded in the stomach and could be stably delivered to the small intestine to benefit intestinal and dermatic health.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.33 no.2
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• pp.93-97
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• 2007

Even in the moderately concentrated anionic surfactant system, some special encapsulation method can shield the papain enzyme from proteolytic attacks. The stabilization of enzyme has been a major issue for successful therapies. In this study, we first stabilized an enzyme, papain in the microcapsules by using polyols, polyethyleneglycol (PEG), poly-propyleneglycol (PPG), and PEG-PPG-PEG block copolymer. In the analysis of EDS and CLSM, it was demonstrated that polyols are effectively located in the interface of papain and polymer. Polyols located in the interface had an ability to buffer the external triggers by hydrophobic partitioning, preventing consequently the catalytic activity of papain in the micro-capsules. Second. we introduced multi-layer capsulation methods containing ion complex. Such a moderately concentrated anionic surfactant system as wash-off cleansers, surfactants and waters can cause instability of entrapped enzymes. Surfactants and water in our final products swell the surface of enzyme capsules and penetrate into the core so easily that we can not achieve the effect of enzyme, papain. In this case, the ion complex multi-layer capsule composed of sodium lauroyl sarcosinate and polyquaternium-6 could effectively prevent water from penetration into the core enzyme, followed by in vivo test, and evaluate the stratum corneum (SC) turn-over speed.

• Archives of Pharmacal Research
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• v.29 no.10
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• pp.859-865
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• 2006

With an approach to study the anti-tumor effects and mechanism of selenium compound, we investigated the anti-tumor activity and mechanism of $Na_5SeV_5O_{18}{\cdot}3H_2O$ (NaSeVO) in K562 cells. The results showed that $0.625{\sim}20\;mg/L$ NaSeVO could significantly inhibit the proliferation of K562 cells in vitro in a time- and concentration-dependent manner as determined by microculture tetrazolium (MTT) assay, the IC50 values were 14.41 (4.45-46.60) and 3.45 (2.29-5.22) mg/L after 48 hand 72 h treatment with NaSeVO respectively. In vivo experiments demonstrated that i.p. administration of 5, 10 mg/kg NaSeVO exhibited an significant inhibitory effect on the growth of transplantation tumor sarcoma 180 (S180) and hepatoma 22 (H22) in mice, with inhibition rate 26.8% and 58.4% on S180 and 31.3% and 47.4% on H22, respectively. Cell cycle studies indicated that the proportion of G0/G1 phase was increased at 2.5 mg/L while decreased at 10 mg/L after treatment for 24, 48 h. Whereas S phase was decreased at 2.5-5 mg/L and markedly increased at 10 mg/L after treatment for 48 h. After treatment for 24 h, 10 mg/L NaSeVO also markedly increased S and G2/M phases. Take together, the result clearly showed that NaSeVO markedly increased S and G2/M phases at 10 mg/L. The study of immunocytochemistry showed that the expression bcl-2 is significantly inhibited by 10 mg/L NaSeVO, and bax increased. Morphology observation also revealed typical apoptotic features. NaSeVO also significantly caused the accumulation of $Ca^{2+}$ and $Mg^{2+}$, reactive oxygen species (ROS) and the reduction of pH value and mitochondrial membrane potential in K562 cells as compared with control by confocal laser scanning microscope. These results suggest that NaSeVO has anti-tumor effects and its mechanism is attributed partially to apoptosis induced by the elevation of intracellular $Ca^{2+}$, $Mg^{2+}$ and ROS concentration, and a reduction of pH value and mitochondria membrane potential (MMP).

• The Journal of Korean Academy of Prosthodontics
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• v.39 no.1
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• pp.71-83
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• 2001

The purpose of this study was to compare the effects of various surface treatments by measuring removal torque on bone healing around titanium implants. 40 Screw-shaped cp titanium implants with length of 4mm, outer diameter of 3.75mm, and pitch-height of 0.5mm were used Group 1 was left as machined(control), Group 2 was blasted with $50{\mu}m\;Al_2O_3$, group 3 was blasted and etched in etching solution($NH_4OH : H_2O_2:H_2O= 1 : 1 : 5$) at $90^{\circ}C$ for 1 minute group 4 was blasted and oxidated under pure oxygen at $800^{\circ}C$. The implant surface roughness was analyzed with SEM and CLSM(Confocal Laser Scanning Microscope) and implants were placed in proximal tibial metaphysis of 10 New Zealand White rabbits. After 3 months of healing period, removal torque of each implant was measured to compare bone healing around implant. The results obtained were as follows 1. In SEM view, blasting increased the roughness of the surface, but etching of that rough surface decreased the roughness due to the removal of the tip of the peak. Oxidation also decreased the roughness due to formation of needle-like oxide grains on the implant surface. 2. The Sa value from CLSM was least in the machined group($0.47{\mu}m$), greatest in blasted group($1.25{\mu}m$), and the value decreased after etching($0.91{\mu}m$) and oxidation($0.94{\mu}m$). 3. The removal torque of etched group(24.5Ncm) was greater than that of machined group(16.7Ncm) (P<0.05), and was greatest in the oxidated group(40.3Ncm) and the blasted group(34.7Ncm).

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.301-309
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• 1997

The objective of this study was to determine the microtubule assembly and chromatin configuration during the first cell cycle in bovine oocytes following injection of spermatozoon, sperm head and tail. The microtubule and chromatin configuration was imaged with fluorescent labeled monoclonal ${\alpha}$-tubulin antibody and propidium iodide under laser scanning confocal microscope. Microtubule and chromatin dynamics in bovine oocytes following intracytoplasmic sperm injection (ICSI) were not different from those observed during in vitro fertilization (IVF). Following ICSI, the microtubular aster was observed around sperm midpiece. During pronuclear formation, the sperm aster was enlarged and seen around male and female pronuclei. At mitotic metaphase, the microtubular spindle assemble astral poles and chromosomes were aligned on the spindle equator. At mitosis, asters were concentrated to each spindle pole and they filled the cytoplasm. After injection of the isolated sperm head, the microtubular aster was not seen around sperm head in any cases (0/18). Instead, microtubules were organized from the cytoplasm, which filled the whole cytoplasm during pronuclear apposition. These microtubules seem to move male and female pronuclei. These results suggest that isolated sperm head can develop into normal pronucleus in mature bovine oocytes, and competent to participate syngamy with the ootid chromatin. The functional microtubules following isolated sperm head injection in bovine oocytes appeared to be organized solely from maternal stores.

• Korean Journal of Microbiology
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• v.50 no.4
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• pp.360-367
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• 2014

Candida albicans is an opportunistic and the most prevalent fungal pathogen that can cause superficial and systemic infections in immunocompromised patients. C. albicans can promote the transition from budding yeast to filamentous form, generating biofilms. Infections associated with C. albicans biofilms are frequently resistant to conventional antifungal therapy. Therefore, the development of more effective antifungal drugs related with biofilm formation is required urgently. The roots of Rheum undulatum have been used for medicinal purposes in Korea and China traditionally. The aim of present study was to evaluate the effect of R. undulatum extract upon preformed biofilms of 12 clinical C. albicans isolates and the antifungal activities. Its effect on preformed biofilms was evaluated using XTT reduction assay, and metabolic activity of all tested strains was reduced significantly ($49.4{\pm}6.0%$) at 0.098 mg/ml R. undulatum. The R. undulatum extract blocked the adhesion of C. albicans biofilms to polystyrene surfaces, and damaged the cell membrane integrity of C. albicans which was analyzed by CFDA, AM, and propidium iodide double staining. It caused cell lysis which was observed by Confocal laser scanning and phase contrast microscope after propidium iodide and neutral red staining, respectively. Membrane permeability was changed as evidenced by crystal violet uptake. The data suggest that R. undulatum inhibits biofilm formation by C. albicans, which can be associated with the damage of the cell membrane integrity, the changes in the membrane permeability and the cell lysis of C. albicans.

• Restorative Dentistry and Endodontics
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• v.25 no.2
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• pp.243-253
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• 2000

The purpose of this in vitro study was to evaluate dentin bonding by two different dentin bonding systems(DBS) using acetone based primer or adhesive [All Bond 2(AB2), One Step(OS)] when they were applied by wet or dry bonding technique. Morphology of resin-dentin interface and hybrid layer thickness(HLT) were investigated using Confocal Laser Scanning Microscope(CLSM) and compared to shear bond strength(SBS). 72 extracted sound human molars were randomly divided into 4 groups of 18 teeth each - Group 1.(AW); AB2 by wet bonding. Group 2(AD); AB2 by dry bonding. Group 3.(OW); OS by wet bonding, Group 4.(OD); OS by dry bonding. In 6 teeth of each group, notch-shaped class V cavities(depth 2mm) were prepared on buccal and lingual surface at the cementoenamel juction(12 cavities per group). To obtain color contrast in CLSM observation, bonding resins of each DBS were mixed with rhodamine B and primer of AB2 was mixed with sodium fluorescein. Prepared teeth of each group were treated with AB2, OS, respectively according to the manufacturer's instructions except for dentin surface moisture treatment after acid etching. In group 1 and 3, after acid etching, excess water was removed with wet tissue(Kimwipes), leaving consistently shiny, visibly hydrated dentin surface. In group 2 and 4, dentin surface was dried for 10 seconds at 1 inch distance. The treated teeth were then packed with composite resin(${\AE}$litefil) and light-cured. 12 microscopic samples($60{\sim}80{\mu}m$ thickness) of each group were obtained after longitudinal section and grinding(Exakt cutting and grinding system). Morphological investigation of resin-dentin interface and HLT measurement using CLSM were done. For measurement of SBS, remaining 12 teeth of each group were flattened occlusally to remove all enamel and grinded to 500 grit SiC(Pedemet Specimen Preparation Equipment). After applying DBS on the exposed dentin surface, composite resin was applied in the shape of cylinder, which has 5mm diameter, 1.5mm thickness, and light cured. SBS was measured using Instron with a crosshead speed of 0.5mm/min. It was concluded as follows, 1. HLT of AW(mean: $2.59{\mu}m$) was thicker than any other group, and followed by AD, OW, OD in descending order(mean; 2.37, 2.28, $1.92{\mu}m$). Only OD had statistically significant differences(p<0.05) to AW and AD. 2. There were intimate contact of resin and dentin at the interface in wet bonding groups, but gaps or irregular interfaces were observed in dry bonding groups. 3. The length, diameter, density of resin tags were various even in the same group without significant differences between groups and lots of adhesive lateral branches were observed. 4. There were no statistically significant difference of SBS between AB2 and OS, but SBS of wet bonding groups were significantly higher(p<0.05) than dry bonding groups. 5. There were no consistent relationships between HLT and SBS.

• Tribology and Lubricants
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• v.39 no.2
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• pp.49-55
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• 2023

Currently, the demand for green technologies toward a sustainable future is rapidly increasing due to growing concern over environmental issues. Methanol is biodegradable and can provide clean combustion to reduce sulfur oxide and nitrogen oxide emissions, and therefore it is a candidate fuel for marine engines. However, the effect of methanol on tribological characteristic degradation should be addressed for methanol-fueled engines. In this study, the methanol addition effects on tribological characteristic degradation is experimentally assessed using a pin-on-disk tribo-tester. Ni-based alloy is used as a target material due to its broad applicability as an engine component material. For a lubricant, engine oil with and without methanol are used. The tests are conducted for up to 10,000 cycles under boundary lubrication while the change in friction force is monitored. Additionally, the wear rate is determined based on laser scanning confocal microscope data. An additional test in which methanol is added at regular intervals is performed with an aim to directly observe its effect on friction. Overall, the friction coefficient increases slightly with increasing methanol concentration. Furthermore, the wear rate of the pin and disk increase significantly with methanol addition. The results also indicate that the friction increases instantaneously with methanol addition at the contacting interface. These findings may be useful for better understanding the methanol effect on the tribological characteristics of Ni-based alloys for methanol-fueled engines with improved performance.

• Journal of Acupuncture Research
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• v.22 no.3
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• pp.155-167
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• 2005

Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Cobrotoxin on binding affinity of cobrotoxin with P50, $IKK{\alpa}$ and $IKK{\beta}$, activities of NF-${\kappa}B$, Cell viability of astrocyte, expressions of protein molecules of NF-${\kappa}B$ such as P50, P-$1{kappa}B$, $1{\kappa}B$ and iflammation related genes such as Cox-2, iNOS, cPLA2 in the SNP or LPS induced Inflammatory pathway of Rats' astrocytes. Methods : In this study, The expression of cytosolic phospholipase A2, Nitric oxcide, Cyclooxygenase-2 and inducible nitrogen oxide synthase was determined by western blotting with corresponding antibodies, and the generation of NF-${\kappa}B$ was assayed by EMSA method in astrocytes of rats. The Cell viability of astrocytes was determined by MTT assay, and Binding affinity of Cobrotoxin with P50, $IKK{\alpha}$ and $IKK{\beta}$ was assayed by Surface plasmon resonance analysis, and NF-${\kappa}B$ dependent luciferase activity was determined by luciferase analysis, and Uptake of cobrotoxin in astrocytes was identified by Confocal laser scanning microscope Results : 1. Compared with control, LPS-induced NF-${\kappa}B$ DNA binding activity was decreased significantly by 0.1, $0.5{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 2. Compared with control, LPS-induced NF-kB dependent luciferase expression was decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 3. Compared with control, SNP induced P50, $I{\kappa}B$ expressions in astrocyte were decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin and P-$1{\kappa}B$ expression was decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 4. Compared with control, LPS induced P50, $1{\kappa}B$ expressions in astrocyte were decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 5. Compared with control, SNP induced Cox-2, iNOS, CPLA2 expressions in astrocyte were decreased significantly by $1{\mu}g/m{\ell}$ of Cobrotoxin. 6. Compared with control, LPS induced Cox-2, cPLA2 expressions in astrocyte were decreased significantly by 0.1, 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin and iNOS expression was decreased significantly by 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin. 7. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, SNP-induced NF-${\kappa}B$ DNA bindins activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM. 8. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, LPS-induced NF-${\kappa}B$ DNA binding activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM, Cobrotoxin $0.5{\mu}g/m{\ell}$with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM 9. Compared with $0.1{\mu}g/m{\ell}$ of cobrotoxin, SNP induced P50 expressions in astrocyte were increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM. 10. The uptake of the labeled cobrotoxin into the cells was shown under a confocal laser scanning microscope. cobrotoxin was uptaken into the membrane and nucleus of astrocytes. Conclusions : In summary, the present results demonstrate that cobrotoxin directly binds to sulfhydryl group of p50 and IKKS resulting In the reduction of translocation of p50 and IkB release, thereby inhibits activation of NF-${\kappa}B$, and suggest that pico to nanomolar range of cobrotoxin could inhibit the expression of genes in the NF-${\kappa}B$ signal pathway.