• Journal of the Korean Wood Science and Technology
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• 제33권1호통권129호
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• pp.48-57
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• 2005

플라스틱 가소제로 사용되고 있는 프탈레이트류(phthalate esters)의 백색부후균에 의한 분해 특성을 확인하기 위해 본 연구에서는 프탈레이트 제조에 사용되고 있는 전구체로서 프탈산(phthalic acid)의 분해를 Polyporus brumalis를 이용하여 확인하였다. 프탈산(phthalic acid) 50 ppm을 액상 배지에 처리하고 균체에 의한 프탈산의 감소율을 확인한 결과, 배양 4일 후부터 감소하기 시작하여 배양 24일에는 배지 내에서 검출되지 않아 완전히 분해되었음을 확인하였고, 배양 기간 동안 리그닌 분해 효소 활성 변화는 배양 10일 후부터 대조구에 비해 프탈산 처리구의 효소 활성이 저조하게 나타남을 확인하였다. Esterase의 경우에는 대조구에서 보여지는 효소 활성 변화와는 달리 배양 10일 이후에 지속적으로 증가하는 것을 확인하였다. 영양원으로 사용된 glucose는 배양 6일 째까지는 대조구와 프탈산 처리구간에 감소율이 유사하였으나, 이후 배양 시간이 연장됨에 따라 배지 내에서 급격하게 감소하는 것을 확인하여 대조구에 비해 프탈산 처리구가 glucose 소모율이 낮았다. 이는 프탈산이 균체에 흡수된 후에 대사작용에 이용되기 때문이라 사료된다. 또한, 균체외 효소에 의한 프탈산의 화학적 구조변화를 GC/MS를 이용하여 분석한 결과 반응 12시간 이내에 무수프탈산으로 탈수 반응이 급격히 진행되는 것을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권9호
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• pp.1381-1388
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• 2019

Objective: The aims of this study were to reveal the magnitude of the differences in protein structures at a cellular level as well as protein utilization and availability among soybean meal (SBM), canola meal (CM), and rapeseed meal (RSM) as feedstocks in China. Methods: Experiments were designed to compare the three different types of feedstocks in terms of: i) protein chemical profiles; ii) protein fractions partitioned according to Cornell Net Carbohydrate and Protein System; iii) protein molecular structures and protein second structures; iv) special protein compounds-amino acid (AA); v) total digestible protein and energy values; vi) in situ rumen protein degradability and intestinal digestibility. The protein second structures were measured using FT/IR molecular spectroscopy technique. A summary chemical approach in National Research Council (NRC) model was applied to analyze truly digestible protein. Results: The results showed significant differences in both protein nutritional profiles and protein structure parameters in terms of ${\alpha}-helix$, ${\beta}-sheet$ spectral intensity and their ratio, and amide I, amide II spectral intensity and their ratio among SBM, CM, and RSM. SBM had higher crude protein (CP) and AA content than CM and RSM. For dry matter (DM), SBM, and CM had a higher DM content compared with RSM (p<0.05), whereas no statistical significance was found between SBM and CM (p = 0.28). Effective degradability of CP and DM did not demonstrate significant differences among the three groups (p>0.05). Intestinal digestibility of rumen undegradable protein measured by three-step in vitro method showed that there was significant difference (p = 0.05) among SBM, CM, and RSM, which SBM was the highest and RSM was the lowest with CM in between. NRC modeling results showed that digestible CP content in SBM was significantly higher than that of CM and RSM (p<0.05). Conclusion: This study suggested that SBM and CM contained similar protein value and availability for dairy cattle, while RSM had the lowest protein quality and utilization.

• Parasites, Hosts and Diseases
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• 제51권4호
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• pp.413-419
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• 2013

The mainstay therapy against leishmaniasis is still pentavalent antimonial drugs; however, the rate of antimony resistance is increasing in endemic regions such as Iran. Understanding the molecular basis of resistance to antimonials could be helpful to improve treatment strategies. This study aimed to recognize genes involved in antimony resistance of Leishmania tropica field isolates. Sensitive and resistant L. tropica parasites were isolated from anthroponotic cutaneous leishmaniasis patients and drug susceptibility of parasites to meglumine antimoniate (Glucantime$^{(R)}$) was confirmed using in vitro assay. Then, complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) and real-time reverse transcriptase-PCR (RT-PCR) approaches were utilized on mRNAs from resistant and sensitive L. tropica isolates. We identified 2 known genes, ubiquitin implicated in protein degradation and amino acid permease (AAP3) involved in arginine uptake. Also, we identified 1 gene encoding hypothetical protein. Real-time RT-PCR revealed a significant upregulation of ubiquitin (2.54-fold), and AAP3 (2.86-fold) (P<0.05) in resistant isolates compared to sensitive ones. Our results suggest that overexpression of ubiquitin and AAP3 could potentially implicated in natural antimony resistance.

• Journal of Ginseng Research
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• 제48권1호
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• pp.31-39
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• 2024

Background: Ginsenoside Rg3, a primary bioactive component of red ginseng, has anti-cancer effects. However, the effects of Rg3-enriched ginseng extract (Rg3RGE) on apoptosis and autophagy in breast cancer have not yet been investigated. In the present study, we explored the anti-tumor effects of Rg3RGE on breast cancer cells stimulated CoCl₂, a mimetic of the chronic hypoxic response, and determined the operative mechanisms of action. Methods: The inhibitory mechanisms of Rg3RGE on breast cancer cells, such as apoptosis, autophagy and ROS levels, were detected both in vitro. To determine the anti-cancer effects of Rg3RGE in vivo, the cancer xenograft model was used. Results: Rg3RGE suppressed CoCl₂-induced spheroid formation and cell viability in 3D culture of breast cancer cells. Rg3RGE promoted apoptosis by increasing cleaved caspase 3 and cleaved PARP and decreasing Bcl2 under the hypoxia mimetic conditions. Further, we identified that Rg3RGE promoted apoptosis by inhibiting lysosomal degradation of autophagosome contents in CoCl₂-induced autophagy. We further identified that Rg3RGE-induced apoptotic cell death and autophagy inhibition was mediated by increased intracellular ROS levels. Similarly, in the in vivo xenograft model, Rg3RGE induced apoptosis and inhibited cell proliferation and autophagy. Conclusion: Rg3RGE-stimulated ROS production promotes apoptosis and inhibits protective autophagy under hypoxic conditions. Autophagosome accumulation is critical to the apoptotic effects of Rg3RGE. The in vivo findings also demonstrate that Rg3RGE inhibits breast cancer cell growth, suggesting that Rg3RGE has potential as potential as a putative breast cancer therapeutic.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3723-3727
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• 2015

The p53 tumor suppressor protein is a principal mediator of growth arrest, senescence, and apoptosis in response to a broad array of cellular damage. p53 is a substrate for the ubiquitin-proteasome system, however, the ubiquitin-conjugating enzymes (E2s) involved in p53 ubiquitination have not been well studied. UBE2Q1 is a novel E2 ubiquitin conjugating enzyme gene. Here, we investigated the effect of UBE2Q1 overexpression on the level of p53 in the MDA-MB-468 breast cancer cell line as well as the interaction between UBE2Q1 and p53. By using a lipofection method, the p53 mutated breast cancer cell line, MDA-MB-468, was transfected with the vector pCMV6-AN-GFP, containing UBE2Q1 ORF. Western blot analysis was employed to verify the overexpression of UBE2Q1 in MDA-MB-468 cells and to evaluate the expression level of p53 before and after cell transfection. Immunoprecipitation and GST pull-down protocols were used to investigate the binding of UBE2Q1 to p53. We established MDA-MB-468 cells that transiently expressed a GFP fusion proteins containing UBE2Q1 (GFP-UBE2Q1). Western blot analysis revealed that levels of p53 were markedly lower in UBE2Q1 transfected MDA-MB-468 cells as compared with control MDA-MB-468 cells. Both in vivo and in vitro data showed that UBE2Q1 co-precipitated with p53 protein. Our data for the first time showed that overexpression of UBE2Q1can lead to the repression of p53 in MDA-MB-468 cells. This repression of p53 may be due to its UBE2Q1 mediated ubiquitination and subsequent proteasome degradation, a process that may involve direct interaction of UBE2Q1with p53.

• 생명과학회지
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• 제17권4호
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• pp.552-560
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• 2007

본 연구에서는 인체백혈병세포 U937에서 HDAC 저해제에 의한 증식억제, 세포주기 교란 및 apoptosis 유도에 미치는 Bcl-2 유전자의 영향에 관하여 조사하였다. 이를 위하여 U937/vector 및 U937/Bcl-2 세포주를 대상으로 대표적인 HDAC 저해제인 TSA 및 Na-B 처리에 의한 세포 증식 및 생존율에 미치는 영향을 조사한 결과, TSA에 의한 U937 세포의 증식억제 및 생존율의 감소는 Bcl-2의 과발현에 의하여 차단되는 효과를 보였으나, Na-B는 U937/vector 및 U937/Bcl-2세포사이에 큰 변화를 보이지는 않았다. 세포주기 교란효과에서 Na-B는 TSA에 비하여 유의적인 차이를 보이지 못하였으며, 이는 TSA에 의한 apoptosis가 U937/Bcl-2 세포에서는 억제되었으나, Na-B에 의한 apoptosis는 Bcl-2의 과발현에 의하여 차단되지 못한 것과 연관성이 있는 결과였다. 또한 TSA에 의한 apoptosis 유발의 Bcl-2에 의한 차단 효과는 TSA에 의하여 활성화된 caspase의 활성 억제, Bcl-2 발현 자체의 완화 등 apoptosis 조절 인자들의 발현 및 활성 변화에 기인 된 것임을 알 수 있었다.

• 대한치과보철학회지
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• 제45권5호
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• pp.611-620
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• 2007

Statement of problem: Second-generation indirect composite resins have been improved flexural strength, compressive strength, hydrolytic degradation resistance, wear resistance compared to first-generation indirect composite resins, but there are still some problems as hydrolysis and low wear resistance. Some manufacturers claim that wear resistance of their materials has been improved, but little independent study has been published on wear properties of these materials and the properties specified in the advertising materials are largely derived from in-house or contracted testing. Purpose: This study was to evaluate the wear of indirect composite resins (SR Adore, Sinfony, Tescera ATL) and gold alloy against the human enamel. Material and method: Extracted human incisors and premolars were sectioned to $2{\times}2{\times}2mm$ cube and embedded in the clear resin and formed conical shaped antagonist to fit the jig of pin-on-disk tribometer. Total 20 antagonists were stored in distilled water. Five disk samples, 24mm in diameter and 1.5mm thick, were made for each of three groups of indirect composite resins and gold alloy group, and polished to #2,000 SiC paper on auto-polishing machine. Disk specimens were tested for wear against enamel antagonists. Wear test were conducted in distilled water using a pin-on-disk tribometer under condition (sliding speed 200rpm contact load 24N, sliding distance 160m). The wear of the enamel was determined by weighing the enamel antagonist before and after test, and the weight was converted to volumes by average density. The wear tracks were analyzed by scanning electron microscopy and surface profilometer to elucidate the wear mechanisms. Statistical analysis of the enamel wear volume, wear track depth and wear tract width of disk specimens were accomplished with one-way ANOVA and the means were compared for significant differences with Scheffe's test. Results: 1. The enamel wear was most in gold alloy, but there were no statistically significant differences among all the groups (P>.05). 2. In indirect composite resin groups, the group to make the most shallow depth of wear tract was Sinfony, followed by Tescera ATL, SR Adoro (P<.05). Gold alloy was shallower than Sinfony, but there was no statistically significant difference between Sinfony and gold alloy (P>.05). 3. The width of wear tract of SR Adore was larger than the other groups (P<.05), and there were no statistically significant differences among the other groups (P>.05). 4. SEM analysis revealed that Sinfony and gold alloy showed less wear scars after test, Tescera ATL showed more wear scars and SR Adore showed the most. Conclusion: Within the limits of this study, Sinfony and gold alloy showed the least wear rates and showed similar wear patterns.

• Toxicological Research
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• 제21권1호
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• pp.31-37
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• 2005

The inhibitory activity against bisphenol-A (BPA), one of well-known endocrine disrupters was examined with the water extracts prepared from the Stem Bark of Syringa velutina (SBS). In this study, we have investigated the effect of SBS on the toxicity caused by BPA in human breast cancer cell line, MCF-7 cells and immature Sprague-Dawley rats. In the estrogen receptor-mediated proliferation assay using MCF-7 cells, BPA (16 ng/ml) induced the cell proliferation, but the water extract of SBS inhibited BPA-induced cell proliferation in a dose-dependent manner. These results are associated with PARP degradation and specific cleavage of anti-apoptotic protein Bcl-2 of apoptotic regulatory factors. Additionally, the BPA (400 mg/100 g) significantly induced the increase of the uterine and virginal weights, while SBS (50 mg/100 g) showed the inhibitory action against BPA, i.e. caused the increase of estrogen-related organ weights in immature rat uterotrophic assay. Taken together, the present data suggest that SBS may have anti-toxicity activities against BPA in vitro and in vivo systems. SBS may be capable of inhibiting adverse effects of BPA such as reproductive disorder.

• 생명과학회지
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• 제23권8호
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• pp.1057-1063
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• 2013

진피(Citri Pericarpium)의 항암작용 기전 해석을 위하여 U937 백혈병 세포의 apoptosis 유발에 미치는 메탄올 추출물(EMCP)의 영향을 조사하였으며, apoptosis 조절에 중요한 몇 가지 유전자들의 발현 및 활성 변화, ROS의 생성 변화를 조사하였다. EMCP 처리에 의한 U937 세포의 증식 억제는 apoptosis 유도와 연관성이 있음을 DAPI 염색을 통한 apoptotic body 출현의 증가 및 Flow cytometry 분석에 의한 Sub-G1기 세포 빈도의 증가로 확인하였다. EMCP 처리에 의한 apoptosis 유도에서 Bax 발현 증가, caspases의 활성 및 PARP의 단편화 등이 동반되었으며, ROS 생성의 증가와 연관성이 있었다. 산화적 손상에 대해 세포나 조직을 보호하는 역할을 하는 것으로 알려진 세포 내 항산화 효소인 HO-1의 발현이 EMCP의 처리에 의해 증가되었으나 ROS 생성 억제제인 NAC의 전처리에 의해 감소된 HO-1의 발현은 전사인자인 Nrf2의 핵으로의 이동 억제와 관련되어 있었다. 이상의 결과에서 EMCP 처리에 의한 U937 세포의 apoptosis 유발에는 ROS 생성의 증가와 pNrf2에 의해 조절되는 HO-1의 발현 증가가 중요한 기전으로 작용한다는 것을 알 수 있었다. 이러한 자료는 진피의 항암기전 해석을 이해하는데 중요한 기초자료로서 활용될 수 있을 것으로 생각된다.

• 생명과학회지
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• 제25권1호
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• pp.1-7
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• 2015

칼슘 의존적으로 활성화되는 neutral protease calpain에 의한 단백질 분해는 세포의 성장을 조절하는데 중요한 단백질들의 역할에 매우 중요한 역할을 한다. Cyclin의 분해는 세포주기의 진행을 위한 필연적인 과정이다. D-type cyclins는 외부자극이나 신호에 의하여 세포주기의 G1 초기에 합성이 된 후 cyclin-dependent kinases (cdk4 및 cdk6)와의 결합하여 세포주기 S기 진입을 촉진하는 역할을 한다. 본 연구에서는 cyclin D3 단백질이 calpain protease에 의하여 번역 후 수준에서 조절 받고 있음을 제시하였다. 본 실험의 조건에서 lovastatin과 actinomycin D가 처리된 PC-3-M 전립선 암세포에서 cyclin D3 단백질의 발현이 완전히 사라졌지만, calpain inhibitor인 LLnL의 처리에 의하여 정상 수준으로 회복되었음을 알 수 있었다. 그러나 26S proteasome의 선택적 억제제인 lactacystin, lysosome 억제제인 ammonium chloride 및 chloroquine, serine protease 억제제인 PMSF는 동일 조건에서 lovastatin과 actinomycin D 처리에 의한 cyclin D3 단백질의 발현저하를 억제하지는 못하였다. In vitro 조건에서 순수 분리된 calpain은 cyclin D3 단백질을 칼슘 농도 의존적으로 분해하였으며, cyclin D3 단백질의 반감기는 LLnL 처리에 의하여 매우 유의적으로 증가되었다. 또한 calpain 저해인자인 calpastatin의 과발현은 PC-3-M 세포에서 뿐만 아니라 NIH 3T3 섬유아세포에서도 cyclin D3 단백질의 반감기 및 안전성을 증대시켰다. 이러한 결과는 cyclin D3 단백질이 칼슘에 의해 활성화 되는 protease calpain에 의해 조절됨을 보여주는 것이다.