• 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.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 국제심포지움
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• pp.59-72
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• 2002

The anticarcinogenic effects of epicatechin(EC) and ginsenoside Rb2(Rb2), which are major components of green tea and Korea ginsen, respectively, were investigated using a model system of gap junctional intercellular communication (GJIC) in WB-F344 rat liver epithelial cells. 12-O-Tetradecanoylphorbol-13-accetate (TPA) and hydrogen preoxide, known as cancer promoters, inhibited GJIC in the epithelial cells as determined by the scrape loading/dye transfer assay, fluorescence redistribution assay after photobleaching, and immunofluorescent staining of connexin 43 using a laser confocal microscope. The inhibition of GJIC by TPA and H2O2 was prevented with treatment of Rb2 or Ec. The effect of EC on GJIC was stronger in TPA-treated cells than in H2O2-treated cells, while the effect of Rb2 was opoosite to that of EC. EC, at the concentration of 27.8$\mu$g/ml, prevented the TPA-induced GJIC inhibition by about 60%. Rb2, at the concentration of 277$\mu$g/ml, recovered the H2O2-induced GJIC inhibition by about 60%. These results suggest that Rb2 and EC may prevent human cancers by preventing the down-regulation of GJIC during the cancer promotion phase and that the anticancer effect of green tea and Korea ginseng may come from the major respective conponents, EC and Rb2.

• Bulletin of the Korean Chemical Society
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• 제35권8호
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• pp.2290-2294
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• 2014

For the present study, rhEGF was encapsulated into solid lipid nanoparticles (SLNs). The SLNs were prepared by the $W_1/O/W_2$ double emulsification method combined with the high pressure homogenization method and the physical properties such as particle size, zeta-potential and encapsulation efficiency were measured. The overall particle morphology of SLNs was investigated using a transmission electron microscopy (TEM). The percutaneous skin permeation and accumulation property of rhEGF was evaluated using Franz diffusion cell system along with confocal laser scanning microscopy (CLSM). The mean particle size of rhEGF-loaded SLNs was $104.00{\pm}3.99nm$ and the zeta-potential value was in the range of -$36.99{\pm}0.54mV$, providing a good colloidal stability. The TEM image revealed a spherical shape of SLNs about 100 nm and the encapsulation efficiency was $18.47{\pm}0.22%$. The skin accumulation of rhEGF was enhanced by SLNs. CLSM image analysis provided that the rhEGF rat skin accumulation is facilitated by an entry of SLNs through the pores of skin.

• Toxicological Research
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• 제31권1호
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• pp.11-16
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• 2015

Our body's immune system has defense mechanisms against pathogens such as viruses and bacteria. Immune responses are primarily initiated by the activation of toll-like receptors (TLRs). In particular, TLR4 is well-characterized and is known to be activated by gram-negative bacteria and tissue damage signals. TLR4 requires myeloid differentiation factor 2 (MD2) as a co-receptor to recognize its ligand, lipopolysaccharides (LPS), which is an extracellular membrane component of gram-negative bacteria. Gambogic acid is a xanthonoid isolated from brownish or orange resin extracted from Garcinia hanburyi. Its primary effect is tumor suppression. Since inflammatory responses are related to the development of cancer, we hypothesized that gambogic acid may regulate TLR4 activation. Our results demonstrated that gambogic acid decreased the expression of pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6, IL-12, and $IL-1{\beta}$) in both mRNA and protein levels in bone marrow-derived primary macrophages after stimulation with LPS. Gambogic acid did not inhibit the activation of Interferon regulatory factor 3 (IRF3) induced by TBK1 overexpression in a luciferase reporter gene assay using IFN-${\beta}$-PRD III-I-luc. An in vitro kinase assay using recombinant TBK1 revealed that gambogic acid did not directly inhibit TBK1 kinase activity, and instead suppressed the binding of LPS to MD2, as determined by an in vitro binding assay and confocal microscopy analysis. Together, our results demonstrate that gambogic acid disrupts LPS interaction with the TLR4/MD2 complex, the novel mechanism by which it suppresses TLR4 activation.

• The Plant Pathology Journal
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• 제20권2호
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• pp.97-102
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• 2004

Paenibacillus polymyxa E681, with its plant growth promotion and root colonization ability, has been proven to be a promising biocontrol agent of cucumber and barley. This study investigated the attributes related to the movement of bacteria from the seed to the radicle and to the whole root system. It also illustrated the existing form and population changes of the bacteria on seed and root using the scanning electron microscope and confocal laser scanning microscopy. The bacteria invaded and colonized the inside of the seed coat while the seeds were soaked in bacterial suspension. Almost the same number of bacteria on seed surface invaded the inside of the seed coat right after seed soaking. The population densities of E681 increased greatly inside as well as on the surface of the seed before the radicle emerged. The bacteria attached on the emerging radicle directly affected the initial population of newly emerg-ing root. The colonized cells on the root were arranged linearly toward the elongation of the root axis. In addition to colonizing the root surface, strain E681 was found inside the roots, where cells colonized the inter-cellular space between certain epidermal and cortical cells. When the cucumber seeds were soaked in bacterial suspension and sown in pot, the bacterial populations attached on both the surface and inside of the root were sustained up to harvesting time. This means that E681 successfully colonized the root of cucumber and sustained its population density up to harvesting time through seed treatment.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1369-1376
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• 2006

Self-organized nanogels were prepared from pullulan/biotin conjugates (PU/Bio) for the development of an effective anticancer drug delivery system. The degree of biotin substitution was 11, 19, and 24 biotin groups per 100 anhydroglucose units of pullulan. The physicochemical properties of the nanogels (PU/Bio1, 2 and 3) in aqueous media were characterized by dynamic light scattering, transmission electron microscopy, and fluorescence spectroscopy. The mean diameter of all the samples was less than 300 nm with a unimodal size distribution. The critical aggregation concentrations (CACs) of the nanoparticles in distilled water were $2.8{\times}10^{-2},\;1.6{\times}10^{-2}$, and $0.7{\times}10^{-2}mg/ml$ for the PU/Bio1, 2, and 3, respectively. The aggregation behavior of the nanogels indicated that biotin can perform as a hydrophobic moiety. To observe the specific interaction with a hepatic carcinoma cell line (HepG2), the conjugates were labeled with rhodamine B isothiocyanate (RITC) and their intensities measured using a fluorescence microplate reader. The HepG2 cells treated with the fluorescence-labeled PU/Bio nanoparticles were strongly luminated compared with the control (pullulan). Confocal laser microscopy also confirmed internalization of the PU/Bio nanogels into the cancer cells. Such results demonstrated that the biotin in the conjugate acted as both a hydrophobic moiety for self-assembly and a tumor-targeting moiety for specific interaction with tumor cells. Consequently, PU/Bio nanogels would appear to be a useful drug carrier for the treatment of liver cancer.

• Restorative Dentistry and Endodontics
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• 제41권2호
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• pp.114-120
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• 2016

Objectives: The aim of this study was to compare the push-out bond strength and dentinal tubule penetration of root canal sealers used with coated core materials and conventional gutta-percha. Materials and Methods: A total of 72 single-rooted human mandibular incisors were instrumented with NiTi rotary files with irrigation of 2.5% NaOCl. The smear layer was removed with 17% ethylenediaminetetraacetic acid (EDTA). Specimens were assigned into four groups according to the obturation system: Group 1, EndoRez (Ultradent Product Inc.); Group 2, Activ GP (Brasseler); Group 3, SmartSeal (DFRP Ltd. Villa Farm); Group 4, AH 26 (Dentsply de Trey)/gutta-percha (GP). For push-out bond strength measurement, two horizontal slices were obtained from each specimen (n = 20). To compare dentinal tubule penetration, remaining 32 roots assigned to 4 groups as above were obturated with 0.1% Rhodamine B labeled sealers. One horizontal slice was obtained from the middle third of each specimen (n = 8) and scanned under confocal laser scanning electron microscope. Tubule penetration area, depth, and percentage were measured. Kruskall-Wallis test was used for statistical analysis. Results: EndoRez showed significantly lower push-out bond strength than the others (p < 0.05). No significant difference was found amongst the groups in terms of percentage of sealer penetration. SmartSeal showed the least penetration than the others (p < 0.05). Conclusions: The bond strength and sealer penetration of resin-and glass ionomer-based sealers used with coated core was not superior to resin-based sealer used with conventional GP. Dentinal tubule penetration has limited effect on bond strength. The use of conventional GP with sealer seems to be sufficient in terms of push-out bond strength.

• Korean Chemical Engineering Research
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• 제56권6호
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• pp.826-831
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• 2018

본 연구는 다중 형광이 표지된 이중 분획 입자의 제조에 관한 것이다. 입자 내에서 형광 발현을 분획화하기 위하여, 형광의 여기 및 방출 스펙트럼의 중첩이 적은 두가지의 형광 염료를 선정한다. 또한, 형광 안정성을 확보하기 위하여 선정된 형광 염료는 입자를 구성하는 소재와 함께 가교될 수 있도록 분자 내에 아크릴레이트(acrylate) 작용기를 포함한다. 공초점 현미경 촬영을 통하여 선정된 형광 물질을 이용하여 제조된 입자에서 강한 형광 발현 및 형광의 분획화를 확인하였다. 더 나아가 4주 동안 형광 발현 및 세기를 측정하여 장기간의 형광 안정성을 검증하였다. 본 연구에서 제조된 다중 형광 표지된 이중 분획 입자는 다중 표적형 약물 전달 체계, 3차원 브라운 운동의 해석 연구, 3차원의 복잡한 자기 조립체 형상의 규명 연구 등에 널리 활용될 수 있으리라 기대한다.

• The Plant Pathology Journal
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• 제37권2호
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• pp.182-193
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• 2021

The transcription factor SHE1 was identified as an interacting partner with the cucumber mosaic virus (CMV) 1a protein in the yeast two-hybrid system, by a pull-down assay, and via bimolecular fluorescent complementation. Using fluorescent-tagged proteins and confocal microscopy, the CMV 1a protein itself was found distributed predominantly between the nucleus and the tonoplast membrane, although it was also found in speckles in the cytoplasm. The SHE1 protein was localized in the nucleus, but in the presence of the CMV 1a protein was partitioned between the nucleus and the tonoplast membrane. SHE1 expression was induced by infection of tobacco with four tested viruses: CMV, tobacco mosaic virus, potato virus X and potato virus Y. Transgenic tobacco expressing the CMV 1a protein showed constitutive expression of SHE1, indicating that the CMV 1a protein may be responsible for its induction. However, previously, such plants also were shown to have less resistance to local and systemic movement of tobacco mosaic virus (TMV) expressing the green fluorescent protein, suggesting that the CMV 1a protein may act to prevent the function of the SHE1 protein. SHE1 is a member of the AP2/ERF class of transcription factors and is conserved in sequence in several Nicotiana species, although two clades of SHE1 could be discerned, including both different Nicotiana species and cultivars of tobacco, varying by the presence of particular insertions or deletions.

• 공업화학
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• 제34권2호
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• pp.121-125
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• 2023

카세인(casein)은 포유류의 우유에서 발견되는 단백질로 우유에서는 80% 이상 함유되어 있다. 사람의 모유에는 약 20~45%가 포함되어 있으며 생체 적합성이 높아 의료 및 산업 소재로 사용되고 있다. 카세인은 양친매성 구조로 내부는 소수성이기 때문에 수용액에서 마이셀로 자가 조립이 가능하여 난용성 약물을 봉입할 수 있다. 또한, 단백질 고분자 소재로 생분해성을 갖고 있어 약물의 전달체로서 적합한 특징을 가진다. 본 연구에서는 칼슘 이온 외에 마그네슘, 아연, 철 등 생체 내 존재하는 다양한 금속 이온들을 사용하여 각각 효과적인 카세인 나노입자 형성 조건을 규명하였다. 동적 광산란 측정기와 제타 전위 측정을 통해 150 nm 이하의 균일한 사이즈를 유지하고 음전하를 띠는 나노입자가 형성됨을 확인하였다. 또한, 각각의 카세인 나노입자가 HeLa 세포주에서 80% 이상의 생존율을 나타내 낮은 세포 독성을 확인하였고, 카세인 나노입자 내부에 시험 약물로서 나일 레드를 봉입하여 세포 내부로 효과적으로 유입됨을 공초점 현미경으로 입증하였다. 본 실험들을 통해 제조된 카세인 나노입자의 약물 전달체로서의 가능성을 확인하였다.