• Journal of Life Science
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• v.20 no.9
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• pp.1324-1331
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• 2010

Mifepristone (MIF) and Tamoxifen (TAM) have been used in the treatment of prostate cancer and breast cancer for more than a decade. MIF can induce apoptosis in both AR-positive and negative prostate cancer cells. Because of its pleiotropic ligand-receptor properties, TAM exerts cytotoxic activity in estrogen (ER)-positive and various ER.negative cancer cells. However, the molecular mechanisms of these two substances are not yet clear. In the present work, we report that the cytotoxic effects of MIF and TAM are due to the modulation of intracellular $Ca^{2+}$ level in DU-145, androgen-insensitive cells. When the cells were treated with micromolar concentrations of either MIF or TAM, the growth and viability were significantly decreased in a dose- and time-dependent manner. The apoptosis induced by MIF or TAM was further proved and analyzed by confocal laser scanning microscopy (CLSM) and fluorescence-activated cell sorting (FACS). In the cells cultivated in a normal 1.5 mM $Ca^{2+}$ medium, both MIF and TAM also induced an increase of the intracellular $Ca^{2+}$ level in a dose-dependent fashion. Since a change in calcium level could not be found in cells of the $Ca^{2+}$-free medium, the increase of intracellular $Ca^{2+}$ level might be due to an increase in extracellular calcium uptake. Our results show that the apoptotic effect was more prominent in TAM treatment compared to MIF treatment in DU-145 cells. The above findings might be due to the difference in the uppermost pathways of apoptosis induced by either MIF or TAM. When we checked the level of procaspase-8 activation, TAM showed minor level of activation, as opposed to MIF, which exerted strong activation. In both treatments, the levels of anti-apoptotic protein Bcl-2 decreased, and pro-apoptotic protein Bax level increased more than 2-fold. The activation of caspase-3, a key protease enzyme in the downstream pathway of apoptosis, was much higher in the cells treated with TAM, compared to the MIF treatment. The overall apoptotic activity shown in the present work was closely related to intracellular $Ca^{2+}$ concentration levels. Therefore, the cytotoxic activity induced by MIF and TAM might have been due to intracellular calcium modulation.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.9
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• pp.1071-1080
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• 2017

This study examined the adhesiveness of beneficial intestinal bacteria to whole-grains using confocal scanning laser microscopy (CLSM), to demonstrate the prebiotic effects of whole-grains, and to develop prebiotic puffed snacks with these whole-grains. CLSM has been used to observe the adhesiveness of Lactobacillus acidophilus, which belongs to beneficial intestinal bacteria, to whole-grain powders using optical sectioning techniques. The enhanced effects on the growth of beneficial intestinal bacteria with the hot water grain extract were verified using an indirect count method. Finally, a puffed snack was produced with the prebiotic effect and the quality was evaluated by checking the chromaticity and degree of hardness. As a result, L. acidophilus exhibited adhesive ability to whole-grain powders and growth of selected beneficial intestinal bacteria were improved significantly. The Hunter L value of the developed puffed snack increased when seasoning was added. The hardness of the puffed snack with seasoning was higher than that of the control. The results of a sensory evaluation showed that the puffed snack with seasoning was highly rated in the overall preference compared to the control.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.10
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• pp.6007-6012
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• 2013

ANXA2, a member of the annexin family, is overexpressed and plays important roles in tumor development. However, the significance of ANXA2 expression in gastric carcinoma has not been clarified.To elucidate its roles in growth of gastric cancer, ANXA2 expression in SGC-7901 cells was inhibited with a designated siRNA, then cell proliferation, cell cycling, apoptosis and motility were determined by MTT assay, flow cytometry, Hoechst 33342 staining and wound healing assay, respectively. To further assess the behavior of ANXA2 deleted SGC-7901 cells, changes of microstructures were observed under fluorescence microscopy, laser scanning confocal microscopy and electron microscopy. We found that inhibition of ANXA2 expression caused cell proliferation to decrease significantly with G1 arrest, motility to be reduced with changes in pseudopodia/filopodia structure and F-actin and ${\beta}$-tubulin expression, and apoptosis to be enhanced albeit without significance. At the same time, ANXA2 deletion resulted in fewer pseudopodia/filopodia, non-stained areas were increased, contact inhibition among cells reappeared, and expression of F-actin and ${\beta}$-tubulin was decreased, with induction of polymerized disassembled forms. Taken together, these data suggest that ANXA2 overexpression is important to maintain the malignancy of cancer cells, and this member of the annexin family has potential to be considered as a target for the gene therapy of gastric carcinoma.

• Journal of Dental Rehabilitation and Applied Science
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• v.28 no.4
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• pp.339-347
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• 2012

In previous studies, methods for enhancing cellular response on the Hydroxyapatite coated implant surface were described. In this study, the changes of surface characteristics such as surface roughness, contact angle, surface energy and surface morphology were observed when Hydroxyapatite coated Ti discs were immersed in NaCl solution for various time. Hydroxyapatite coated Ti discs were immersed in 0.9% NaCl solution for 7, 14 and 21 days at $37^{\circ}C$. The control group comprises dry identical discs not immersed in a solution. (n=3) All discs were dried in air completely and the surface roughness was measured using confocal laser scanning microscopy(CLSM). Static contact angle was recorded by video contact angle analyzer after dropping distilled water on the surface. The surface energy was calculated from contact angles of the three liquids. Surface was observed using a field emission-scanning electron microscope(FE-SEM). As a result, the surface roughness of immersed Hydroxyapatite coated Ti discs increased significantly and the contact angle decreased comparing with control group discs. The surface energy of immersed discs increased except for discs immersed for 14 days.

• Journal of Periodontal and Implant Science
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• v.29 no.2
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• pp.415-433
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• 1999

Implant stability is the key to long-term successful outcome for osseointegrated implants. To evaluate the initial healing response of bone around HA-coated implants without primary bone contact. 21 HA-coated thread type implants(STERI-OSS?) were placed in the femurs of 5 mongrel dogs, about 1-year old. Implants, 8 mm in length and 3.8mm(experimental 1group), 5.0mm(experimental 2group) and 6.0mm(control group) in diameter, were inserted after 3 holes of 6.0mm in diameter and 10mm in depth were prepared in the surgical sites each dog. Implants were supported by only nonresorbable membrane($Teflon^{(R)}$), in order to prevent the ingrowth of upper soft tissue into the gap between bone and implant, and to maintain each implant to be positioned in the center of the drilled hole. 9 implants with different diameters were inserted in 3 dogs for histologic observation, and 12 implants were inserted in 2 dogs for mobility test and removal torque test. Fluorescent dyes were injected for the observation of new bone formation in order of $Terramycin^{(R)}$, Arizarin $Red^{(R)}$, and $Calcein^{(R)}$ at an interval of 2 weeks. 3 dogs were sacrificed for histologic observation at 4, 8, and 12-week after placement. Light microscopy and confocal laser scanning microscopy were used to qualitatively characterize the bone around HA-coated implant. 2 dogs were sacrificed for mobility test($Periotest^{(R)}$, Simens AG, Bensheim, Germany) and removal torque test($Autograph^{(R)}$ AGS-1000D series, Japan) at 8 and 12-week after placement The results were as follows: 1. Histologic observation showed that osseointegration occurred to both control and experimental groups as time lapse, but delayed bone healing was revealed in 3.8mm group (experimental 1group), compared to contrtol group and 5.0mm group (experimental 2group). 2. The mobility test showed that the experimental groups had no distinguishable movement during experimental periods of 8 and 12-week, and there was no difference in mobility depending on the gap between bone and implant, and time lapse. 3. The removal torque forces were increased depended on the gaps decreasing between bone and implant, and time lapse. The results suggest that HA-coated implant without primary bone contact, based on guided bone regeneration could obtain its stability in all experimental groups as time lapse, but bone healing was delayed in experimental group of 3.8mm. And the results suggested that studies on correlationship between mobility test and removal torque test for implant stability would be necessary.

• Journal of Periodontal and Implant Science
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• v.44 no.2
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• pp.79-84
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• 2014

Purpose: While single-species biofilms have been studied extensively, we know notably little regarding multispecies biofilms and their interactions. The purpose of this study was to develop and evaluate an in vitro multispecies dental biofilm model that aimed to mimic the environment of chronic periodontitis. Methods: Streptococcus gordonii KN1, Fusobacterium nucleatum ATCC23726, Aggregatibacter actinomycetemcomitans ATCC33384, and Porphyromonas gingivalis ATCC33277 were used for this experiment. The biofilms were grown on 12-well plates with a round glass slip (12 mm in diameter) with a supply of fresh medium. Four different single-species biofilms and multispecies biofilms with the four bacterial strains listed above were prepared. The biofilms were examined with a confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM). The minimum inhibitory concentrations (MIC) for four different planktonic single-species and multispecies bacteria were determined. The MICs of doxycycline and chlorhexidine for four different single-species biofilms and a multispecies biofilm were also determined. Results: The CLSM and SEM examination revealed that the growth pattern of the multispecies biofilm was similar to those of single-species biofilms. However, the multispecies biofilm became thicker than the single-species biofilms, and networks between bacteria were formed. The MICs of doxycycline and chlorhexidine were higher in the biofilm state than in the planktonic bacteria. The MIC of doxycycline for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis, F. nucleatum, or A. actinomycetemcomitans. The MIC of chlorhexidine for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis or F. nucleatum. Conclusions: To mimic the natural dental biofilm, a multispecies biofilm composed of four bacterial species was grown. The 24-hour multispecies biofilm may be useful as a laboratory dental biofilm model system.

• 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.

• Restorative Dentistry and Endodontics
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• v.36 no.6
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• pp.490-497
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• 2011

Objectives: This study examined the effect of the uncured dentin adhesives on the bond interface between the resin inlay and dentin. Materials and Methods: Dentin surface was exposed in 24 extracted human molars and the teeth were assigned to indirect and direct resin restoration group. For indirect resin groups, exposed dentin surfaces were temporized with provisional resin. The provisional restoration was removed after 1 wk and the teeth were divided further into 4 groups which used dentin adhesives (OptiBond FL, Kerr; One-Step, Bisco) with or without light-curing, respectively (Group OB-C, OB-NC, OS-C and OS-NC). Pre-fabricated resin blocks were cemented on the entire surfaces with resin cement. For the direct resin restoration groups, the dentin surfaces were treated with dentin adhesives (Group OB-D and OS-D), followed by restoring composite resin. After 24 hr, the teeth were assigned to microtensile bond strength (${\mu}TBS$) and confocal laser scanning microscopy (CLSM), respectively. Results: The indirect resin restoration groups showed a lower ${\mu}TBS$ than the direct resin restoration groups. The ${\mu}TBS$ values of the light cured dentin adhesive groups were higher than those of the uncured dentin adhesive groups (p < 0.05). CLSM analysis of the light cured dentin adhesive groups revealed definite and homogenous hybrid layers. However, the uncured dentin adhesive groups showed uncertain or even no hybrid layer. Conclusions: Light-curing of the dentin adhesive prior to the application of the cementing material in luting a resin inlay to dentin resulted in definite, homogenous hybrid layer formation, which may improve the bond strength.

• Journal of the korean academy of Pediatric Dentistry
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• v.47 no.4
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• pp.446-453
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• 2020

The purpose of this study was to evaluate the effect of Indocyanine Green (ICG) and near-infrared (NIR) diode laser on Streptococcus mutans biofilms depending on ICG concentrations. S. mutans biofilms were formed on a Hydroxyapatite disk, and 0.5, 1.0, 2.0, 3.0, 4.0, 5.0 mg/mL ICG solutions dissolved in sterile distilled water and a NIR diode laser having a power of 300 mW and a wavelength of 808 nm were applied to the biofilms. The temperature changes of the biofilm surface according to the concentrations of the ICG solution were measured using a 1-channel thermocouple thermometer. Compared to the control group, in the groups with only the 3.0, 4.0, 5.0 mg/mL ICG solution application, and in the groups with the 1.0, 2.0, 3.0, 4.0, 5.0 mg/mL ICG solution application and light irradiation, a statistically significant decrease in the bacterial counts were observed. The temperature increase according to the concentration of the ICG solutions was 9.53℃, 10.43℃, 11.40℃, 12.10℃, 12.67℃, and 13.63℃ in ICG solutions of 0.5, 1.0, 2.0, 3.0, 4.0, and 5.0 mg/mL respectively. This study presents the potential for clinical application of ICG and NIR diode lasers as a new method for preventing dental caries.

• Journal of the korean academy of Pediatric Dentistry
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• v.44 no.2
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• pp.170-179
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• 2017

The purpose of this study was to compare the cariogenicity of commercially available bovine milk and low-fat milk in a biofilm model using the CDC Biofilm Reactor. Streptococcus mutans ATCC 25175 biofilms were formed on saliva coated bovine enamel slabs in a CDC Biofilm Reactor. Biofilms were exposed three times per day to one of the following materials: commercial whole milk (fat content: 3.4%), low-fat milk (fat content: 1%), or 0.9% NaCl. Medium pH was measured at different time points. After 5 days, biofilms were separated from slabs to evaluate the CFUs. The biofilm thickness was observed by confocal laser-scanning microscopy (CLSM). Enamel slab's demineralization was assessed by measuring surface microhardness before and after the experiment. For microhardness and CFUs assessment, no significant difference was found among the three groups. All groups showed similar pattern of medium pH change and biofilm thickness. Our results showed that there was no difference in the cariogenicity between whole milk and low-fat milk. Both milks were relatively non-cariogenic compared to the control group.