• 한국산학기술학회논문지
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• 제12권1호
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• pp.312-317
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• 2011

캡사이신 채널로 알려진 바닐로이드 수용체 TRPV1 (캡사이신채널, Transient Receptor Potential Vanilloid 1)은 통증발현에서 중요한 역할을 하는 것으로 알려져 있다. 하지만 TRPV1의 활성조절에 관여하는 단백질에 대하여는 알려진 바가 많지 않다. 최근 rat TRPV1과 직접적으로 결합하는 단백질을 탐색하여 mouse Rab11-FIP3 (rab11-family interaction protein 3)가 rat TRPV1과 직접적으로 결합한다는 것이 보고되었다. Rab11은 여러 가지의 세포내 이동에 관여하는 것으로 보고되었다. 그러므로 Rab11-FIP3과의 결합을 통해 TRPV1의 세포막으로의 이동에 관여할 것으로 추측할 수 있다. 본 연구에서는 전에 보고된 연구가 mouse와 rat 이라는 다른 종의 단백질끼리의 결합이기 때문에 같은 종에서의 상호작용을 확인하고 Rab11-FIP3의 TRPV1의 세포막으로의 이동에서의 역할을 알아보고자 현재까지 동정되지 않은 rat의 Rab11-FIP3의 유전자를 GenBank 서열을 바탕으로 rat 뇌의 RNA 로부터 cDNA 를 클로닝하여 유전자를 분리하고 TRPV1 과의 관계를 세포생물학적으로 알아보았다. 연구결과 rat의 Rab11-FIP3는 489개의 아미노산 서열을 가지고 있으며 human과는 80%, mouse와는 90% 이상 아미노산 서열의 상동성을 보였다. 조직별 분포는 심장, 뇌, 간, 콩팥, 정소에서 발현되고 있는 것을 northern blot assay와 western blot assay 로 확인하였다. rat 의 뇌조직에서 TRPV1 과 Rab11-FIP3 단백질이 결합하여 colocalize 하는 것을 면역화학방법으로 확인하였다. 이 결합은 같은 family 의 TRPV2 와는 결합하지 않는 특이적 결합이므로 Rab11-FIP3 가 TRPV1 과 상호작용하여 세포막으로의 이동에 관여할 것이라는 것을 시사한다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권6호
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• pp.880-886
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• 2007

Ensiled or oven-dried green tea by-products (GTB) were evaluated in goats for their nutritive potential as protein feedstuffs based on in vitro and in vivo digestibility. To elucidate the effects of tea tannin on in vitro digestibility, polyethylene glycol (PEG) was used as a tannin binding agent. Both ensiled and dried GTB contained 31.9 to 32.6% of crude protein (CP) on a dry matter (DM) basis. Phenolics and tannins in soybean meal and alfalfa hay were low or not detected, but they were high in both ensiled and dried GTB (7.3-10.1% DM as total extractable tannins). In vitro protein digestibility in the rumen ranked: soybean meal>alfalfa hay cube>ensiled GTB = dried GTB. The protein digestibility post-ruminally of these feedstuffs showed a similar trend to that in the rumen, but the digestibility of ensiled GTB was significantly higher than that of dried GTB. Addition of PEG improved the in vitro protein digestibility of both kinds of GTB in the rumen and post-ruminally, indicating that tannins suppressed the potential protein digestibility of GTB. The increased protein digestibility by PEG addition was not significantly different between ensiled and dried GTB in the rumen, but the percentage increment of ensiled GTB was higher than dried GTB post-ruminally. In the in vivo digestibility trial, ensiled and dried GTB were offered to goats as partial substitutes for soybean meal and alfalfa hay cubes. Offering both GTB to goats as 5-10% on a DM basis did not affect nutrient digestibility, ruminal pH, volatile fatty acids, and ammonia concentration. However, the eating time of the GTB-incorporated diet was longer than that of the basal diet. It took 1.4 and 1.6 times longer than the control diet, to eat the diet completely when GTB silage was offered at 5 and 10% levels, respectively, of the total diet. These results show that ensiled and dried GTB are useful as partial substitutes for soybean meal and alfalfa hay cubes for goats with respect to nutritive value. Because of lessened palatability, it is recommended that GTB be incorporated into the diet at 5% on a DM basis.

• Journal of Microbiology and Biotechnology
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• 제27권6호
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• pp.1138-1149
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• 2017

The use of microalgal biomass is an interesting technology for the removal of heavy metals from aqueous solutions owing to its high metal-binding capacity, but the interactions with bacteria as a strategy for the removal of toxic metals have been poorly studied. The goal of the current research was to investigate the potential of Burkholderia tropica co-immobilized with Chlorella sp. in polyurethane discs for the biosorption of Hg(II) from aqueous solutions and to evaluate the influence of different Hg(II) concentrations (0.041, 1.0, and 10 mg/l) and their exposure to different contact times corresponding to intervals of 1, 2, 4, 8, 16, and 32 h. As expected, microalgal bacterial biomass adhered and grew to form a biofilm on the support. The biosorption data followed pseudo-second-order kinetics, and the adsorption equilibrium was well described by either Langmuir or Freundlich adsorption isotherm, reaching equilibrium from 1 h. In both bacterial and microalgal immobilization systems in the co-immobilization of Chlorella sp. and B. tropica to different concentrations of Hg(II), the kinetics of biosorption of Hg(II) was significantly higher before 60 min of contact time. The highest percentage of biosorption of Hg(II) achieved in the co-immobilization system was 95% at pH 6.4, at 3.6 g of biosorbent, $30{\pm}1^{\circ}C$, and a mercury concentration of 1 mg/l before 60 min of contact time. This study showed that co-immobilization with B. tropica has synergistic effects on biosorption of Hg(II) ions and merits consideration in the design of future strategies for the removal of toxic metals.

• 대한약침학회지
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• 제19권1호
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• pp.37-44
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• 2016

Objectives: This study examined the relative efficacies of a derivative of betulinic acid (dBA) and its poly (lactide-co-glycolide) (PLGA) nano-encapsulated form in A549 lung cancer cells in vivo and in co-mutagen [sodium arsenite (SA) + benzo[a]pyrene (BaP)]-induced lung cancer in mice in vivo. Methods: dBA was loaded with PLGA nanoparticles by using the standard solvent displacement method. The sizes and morphologies of nano-dBA (NdBA) were determined by using transmission electron microscopy (TEM), and their intracellular localization was verified by using confocal microscopy. The binding and interaction of NdBA with calf thymus deoxyribonucleic acid (CT-DNA) as a target were analyzed by using conventional circular dichroism (CD) and melting temperature (Tm) profile data. Apoptotic signalling cascades in vitro and in vivo were studied by using an enzyme-linked immunosorbent assay (ELISA); the ability of NdBA to cross the blood-brain barrier (BBB) was also examined. The stage of cell cycle arrest was confirmed by using a fluorescence-activated cell-sorting (FACS) data analysis. Results: The average size of the nanoparticles was ~ 110 nm. Confocal microscopy images confirmed the presence of NdBA in the cellular cytoplasm. The bio-physical properties of dBA and NdBA ascertained from the CD and the Tm profiles revealed that NdBA had greater interaction with the target DNA than dBA did. Both dBA and NdBA arrested cell proliferation at G0/G1, NdBA showing the greater effect. NdBA also induced a greater degree of cytotoxicity in A549 cells, but it had an insignificant cytotoxic effect in normal L6 cells. The results of flow cytometric, cytogenetial and histopathological studies in mice revealed that NdBA caused less nuclear condensation and DNA damage than dBA did. TEM images showed the presence of NdBA in brain samples of NdBA fed mice, indicating its ability to cross the BBB. Conclusion: Thus, compared to dBA, NdBA appears to have greater chemoprotective potential against lung cancer.

• BMB Reports
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• 제48권11호
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• pp.618-623
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• 2015

FK506 binding protein 12 (FK506BP) is a small peptide with a single FK506BP domain that is involved in suppression of immune response and reactive oxygen species. FK506BP has emerged as a potential drug target for several inflammatory diseases. Here, we examined the protective effects of directly applied cell permeable FK506BP (PEP-1-FK506BP) on corneal alkali burn injury (CAI). In the cornea, there was a significant decrease in the number of cells expressing pro-inflammation, apoptotic, and angiogenic factors such as TNF-α, COX-2, and VEGF. Both corneal opacity and corneal neovascularization (CNV) were significantly decreased in the PEP-1-FK506BP treated group. Our results showed that PEP-1-FK506BP can significantly inhibit alkali burn-induced corneal inflammation in rats, possibly by accelerating corneal wound healing and by reducing the production of angiogenic factors and inflammatory cytokines. These results suggest that PEP-1-FK506BP may be a potential therapeutic agent for CAI.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3667-3671
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• 2015

Invasion and metastasis is the major cause of tumor recurrence, difficulty for cure and low survival rate. Excavating key transcription factors, which can regulate tumor invasion and metastasis, are crucial to the development of therapeutic strategies for cancers. PU.1 is a master hematopoietic transcription factor and a vital regulator in life. Here, we report that, compared to adjacent non-cancerous tissues, expression of PU.1 mRNA in metastatic hepatocellular carcinoma (HCC), but not primary HCC, was significantly down-regulated. In addition, levels of PU.1 mRNA in metastatic hepatoma cell lines MHCC97L and MHCC97H were much lower than in non-metastatic Hep3B cells. Transwell invasion assays after PU.1 siRNA transfection showed that the invasion of hepatoma cell lines was increased markedly by PU.1 knockdown. Oppositely, overexpression of PU.1 suppressed the invasion of these cells. However, knockdown and overexpression of PU.1 did not influence proliferation. Finally, we tried to explore the potential mechanism of PU.1 suppressing hepatoma cell invasion. ChIP-qPCR analysis showed that PU.1 exhibited a high binding capacity with miR-615-5p promoter sequence. Overexpression of PU.1 caused a dramatic increase of pri-, pre- and mature miR-615-5p, as well as a marked decrease of miR-615-5p target gene IGF2. These data indicate that PU.1 inhibits invasion of human HCC through promoting miR-615-5p and suppressing IGF2. These findings improve our understanding of PU.1 regulatory roles and provided a potential target for metastatic HCC diagnosis and therapy.

• Biomolecules & Therapeutics
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• 제18권1호
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• pp.39-47
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• 2010

Brain-derived neurotrophic factor (BDNF) is a neurotrophic factor involved in neuronal differentiation, plasticity, survival and regeneration. BDNF draws massive attention mainly due to the potential as a therapeutic target in neurological diseases such as depression and Alzheimer's disease. In a primary screening for the natural compounds enhancing BDNF release from cultured rat primary cortical neuron, we found that compounds such as baicalein, tanshinone IIa, cinnamic acid, epiberberine, genistein and wogonin among many others increased BDNF release. All the compounds at $0.1{\mu}M$ of concentration barely showed stimulatory effect on BDNF induction, however, their combination (mixture 1; baicalein, tanshinone IIa and cinnamic acid, mixture 2; epiberberine, genistein and wogonin) showed synergistic increase in BDNF release as well as mRNA and protein expression. The level of BDNF expression was comparable to the maximum BDNF stimulation attainable by a positive control oroxylin A ($20{\mu}M$) without cell toxicity as determined by MTT analysis. Both mixtures synergistically increased the phosphorylation of extracellular signal-regulated kinase (ERK) as well as cAMP response element binding protein (CREB), an immediate and essential regulator of BDNF expression. Similar to these results, mixture of these compounds synergistically inhibited the up-regulation of inducible nitric oxide synthase (iNOS) induced by lipopolysaccharide treatments in rat primary astrocytes. These results suggest that the combinatorial treatment of natural compounds in lower concentration might be a useful strategy to obtain sufficient BDNF stimulation in neurological disease condition such as depression, while minimizing potential side effects and toxicity of higher concentration of a single compound.

• 대한본초학회지
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• 제30권1호
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• pp.67-75
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• 2015

Objectives : The aim of this study is to examine the neuroprotective effect of wheat bran extract (WBE) against ${\beta}$-amyloid ($A{\beta}$)-induced apoptotic cell death in SH-SY5Y human neuroblastoma cells and memory impairment in triple transgenic animal model's of Alzheimer's disease (3xTg AD mice). Methods : In SH-SY5Y cells, MTT assay and TUNEL staining were conducted to evaluate the protective effect of WBE against $A{\beta}_{25-35}$-induced neurotoxicity and apoptosis. Alterations in mitochondrial transmembrane potential (MMP), expression of proapoptotic Bax and antiapoptotic Bcl-2 proteins, cleavage of PARP, and brain-derived neurotrophic factor (BDNF) levels were analyzed to elucidate the neuroprotective mechanism of WBE. To further investigate the memory enhancing effect of WBE, Morris water maze test was performed in 3xTg AD mice. Results : In SH-SY5Y cells, WBE protected against $A{\beta}_{25-35}$-caused cytotoxicity and apoptosis as shown by the restoration of cell viability in MTT assay and inhibition of DNA fragmentation in TUNEL staining. $A{\beta}_{25-35}$-induced apoptotic signals such as dissipation of MMP, decreased Bcl-2/Bax ratio, and cleavage of PARP were suppressed by WBE. Moreover, WBE up-regulated the protein levels of BDNF, which seemed to be mediated by activation of cAMP response element-binding protein (CREB). In 3xTg AD mice, oral administration of WBE attenuated learning and memory deficit as verified by reduced mean escape latency in water maze test. Conclusions : WBE protects neuronal cells from $A{\beta}_{25-35}$-induced apoptotic cell death and restores learning and memory impairments in 3xTg AD mice. These findings suggest that WBE exhibit neuroprotective potential for the management of AD.

• 한국치위생학회지
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• 제9권2호
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• pp.73-88
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• 2009

The purpose of the study was to provide the basic for the prevention of infection control by analyzing the level of recognition and practice of dental hygiene students. It is important for dental clinic staffs to know and practice infection control measures in favor of effective infection control. Thus, this study conducted an one-month questionnaire survey (during October 2008) for dental hygiene students who ever experienced in 16-week clinical practice, and analyzed total 206 questionnaire forms as collected from them, so that it could determine potential associations between their recognition and practice of infection control and thereby suggest an efficient solution for infection control in dental hygiene. 1. It was found that our students' recognition about infection control averaged $4.49{\pm}.58$ points on the whole, and they were best aware of instrumental disinfection and sterilization among other measures, which was followed by hand washing, extirpation control, post-sterilization management, personal protective outfit and instrumental surface maintenance respectively. 2. It was found that our students' practice of infection control averaged $3.85{\pm}.57$ points on the whole, and they practiced hand washing most actively among other measures, which was followed by extirpation control, post-sterilization management, instrumental disinfection and sterilization, personal protective outfit and instrumental surface maintenance respectively. 3. It was found that our students scored higher mean points in recognition about every measure of infection control than those in practice (t=15.676, p=0.000). Particularly, it was notable that there were significant differences between their mean points in recognition and practice of some infection control measures, such as instrumental surface maintenance (t=15.361, p=0.000), personal protective outfit (t=15.245, p=0.000) and instrumental disinfection and sterilization (t=11.169, p=0.000). But there was least significant difference between mean points in recognition and practice of hand washing (t=5.460, p=0.000). 4. For potential associations between recognition and practice of infection control, it was found that higher recognition was in significantly positive associations with higher practice in every measure of infection control (r=.478, p=.000), such as extirpation control (r=.630, p=.000), instrumental disinfection and sterilization (r=.477, p=.000) and post-sterilization management (r=.433, p=.000). 5. It was found that there were differences in our students' recognition depending upon availability of infection control guideline (t=4.587, p=.011), and there were significant differences in practice depending on necessity of infection control education on a statistical basis (t=2.229, p=.027). Overall, it is found that our dental hygiene students are very likely to practice hand washing and extirpation control, because both of these measures are relatively easy to practice or are considered legally binding.

• 한국해양바이오학회지
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• 제2권2호
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• pp.102-107
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• 2007

산업의 발달과 함께 환경오염에 대한 국민적인 관심도는 날로 증가하고 있다. PCB는 우리 주변에 널리 퍼져 있고 먹이사슬을 통해 체내에도 축적되어 인체의 위해성이 우려되는 대표적인 환경오염물질이다. PCB의 노출은 성장기의 두뇌에서 가장 큰 신경독성을 나타내며 영아 및 유아는 상대적으로 높게 노출되어 위험집단으로 분류된다. 본 연구는 PCB의 신경독성에 구조-활성관계가 미치는 영향을 분석하고 PCB에 의한 독성을 최소화 할 수 있는 방안으로서 해양활성물질의 사용가능성을 이해하고자 하였다. PCB노출에 따른 신경세포의 신호전달 체계변화를 분석하기 위하여 Protein Kinase C (PKC)의 변화를 측정하였다. PKC의 전체적인 활성을 [$^3H$]PDBu로 분석한 결과 ortho-position(PCB-105, -123)을 가지고 있는 PCB가 non-ortho (pCB-77, -81) 구조보다 신경에 미치는 영향은 더 높았다. Westem blot 결과 PKC isofonn 중에는 PKC-beta II 및 epsilon의 경우 ortho-position PCB에서 더 높은 활성을 보였다. 이러한 PKC의 변화는 성장기 신경세포에서 신호전달기작의 변화에 많은 영향을 미치므로 이를 예방하거나 차단 할 수 있는 물질을 발견하고자 다양한 키토산을 처리하였다. 그 결과 1백만 달톤 이상의 고분자 키토산의 경우 PCB에 의한 신호전달 기작 변화를 억제할 수 있음을 보였다. 본 연구는 환경오염 등에 의한 독성예방에 키토산의 활용가능성을 제시하였다.