• Toxicological Research
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• 제4권1호
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• pp.37-45
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• 1988

Primary cultures of adult rat hepatocytes were used to study in vitro cytotoxic effects of T-2 toxin on liver cells. When T-2 toxin was added to the culture, a significant depression of the hormonal induction of ${\alpha}$-aminoisobutyric acid (AIB) uptake and tyrosine aminotransferase (TAT) activity was observed. However, T-2 toxin did not affect the uptake of ouabain into hepatocytes. Protein synthesis was inhibited by T-2 toxin, but RNA synthesis was not severely affected. The inhibitory effects of T-2 toxin on protein synthesis was diminished rapidly with culture time and the hepatocytes culture maintained control level of protein synthesis within 24 hrs.

• Bulletin of the Korean Chemical Society
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• 제31권9호
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• pp.2607-2612
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• 2010

Human tissue-type plasminogen activator (tPA) is a valuable thrombolytic agent used to successfully treat acute myocardial infarction, thromboembolic stroke, peripheral arterial occlusion, and venous thromboembolism. Recombinant tPA is accumulated as an inactive form in inclusion bodies of E. coli and is refolded in vitro, which is accompanied by extensive aggregation. In the present study, a tPA protease domain was expressed in an active soluble form in the cytosol of E. coli Rosetta-gami cells, which allowed disulfide bond formation and supplied the tRNA molecules required for six rarely used codons in E. coli. This strategy increased the amount of soluble protease domain protein and avoided the cumbersome refolding process. The purified protease domain not only degraded tPA substrate peptides but also formed a covalently bound complex with plasminogen activator inhibitor-1, as does full-length tPA. Soluble expression and purification of tPA domains may aid in functional analyses of this multi-domain protein, which has been implicated in many physiological and pathological processes.

• 한국동물학회지
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• 제26권3호
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• pp.181-192
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• 1983

이차원 전긱영동법에 의하여 A. proteus의 tD strain과 tD strain이 박테리아와 공생에 의해 유도된 xD strain의 단백질 양성을 비교하였다. Silver stain에 의해 비교 가능한 200여개의 주요 단백질 가운데 tD strain에서 분자량 45,000, 동전점 5.9의 특이성 단백질이, xD strain의 세포액과 공생낭에서는 분자량 29,000, 동전점 5.5의 공생 특이성 단백질이 탐지되었다. 공생 특이성 단백질은 아메바 고은 배양 및 실험 공생 아메바를 이용한 실험 결과 박테리아와 직접 연관 되어 있었다. 탐지된 두 특이성 단백질에 대하여 종전의 세포 핵 이식 및 배양 실험을 통해서 얻어진 결과에 비추어 이들 단백질 상호 연관 및 세포내의 기능에 관하여 논의하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제17권8호
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• pp.1062-1068
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• 2004

Leptin has a major role in the regulation of food intake and energy homeostasis. In addition, leptin participates in many physiological functions including regulation of lipid metabolism. Bovine recombinant leptin protein was produced in E. coli cells in order to understand function of leptin in the regulation of lipid metabolism. The leptin expression vector was constructed in pGEX-4T-3 vector and transformed into E. coli BL21 cells. Expression of the GST-leptin fusion protein was induced with IPTG. The fusion protein was purified using glutathione sepharose 4B batch method, and the recombinant leptin was eluted after thrombin protease digestion. The effect of leptin on glucose transport was examined in the differentiated adipocytes of 3T3-L1 cells. Leptin had no effect on basal and insulin-stimulated glucose transport in 3T3-L1 cells (p>0.05). Effect of recombinant leptin on glucose and acetate transport was examined in adipose tissues of Korean cattle (Hanwoo). Insulin stimulated glucose transport in both intramuscular and subcutaneous adipose tissues (p<0.05), but leptin did not affect glucose transport in both adipose tissues (p>0.05). Insulin stimulated acetate transport in bovine adipose tissues (p<0.05), but leptin did not affect acetate transport (p>0.05). Northern and RT-PCR analyses showed that mRNA levels of uncoupling protein-2 were increased by leptin treatment in 3T3-L1 cells without statistical difference (p>0.05). In conclusion, bovine recombinant leptin did not affect glucose and acetate transport in both 3T3-L1 adipocytes and bovine adipose tissues, while it stimulates UCP-2 mRNA expression in 3T3-L1 cells.

• Nutrition Research and Practice
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• 제9권4호
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• pp.343-349
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• 2015

BACKGROUND/OBJECTIVES: Fermentation of dietary fiber results in production of various short chain fatty acids in the colon. In particular, butyrate is reported to regulate the physical and functional integrity of the normal colonic mucosa by altering mucin gene expression or the number of goblet cells. The objective of this study was to investigate whether butyrate modulates mucin secretion in LS174T human colorectal cells, thereby influencing the adhesion of probiotics such as Lactobacillus and Bifidobacterium strains and subsequently inhibiting pathogenic bacteria such as E. coli. In addition, possible signaling pathways involved in mucin gene regulation induced by butyrate treatment were also investigated. MATERIALS/METHODS: Mucin protein content assay and periodic acid-Schiff (PAS) staining were performed in LS174T cells treated with butyrate at various concentrations. Effects of butyrate on the ability of probiotics to adhere to LS174T cells and their competition with E. coli strains were examined. Real time polymerase chain reaction for mucin gene expression and Taqman array 96-well fast plate-based pathway analysis were performed on butyrate-treated LS174T cells. RESULTS: Treatment with butyrate resulted in a dose-dependent increase in mucin protein contents in LS174T cells with peak effects at 6 or 9 mM, which was further confirmed by PAS staining. Increase in mucin protein contents resulted in elevated adherence of probiotics, which subsequently reduced the adherent ability of E. coli. Treatment with butyrate also increased transcriptional levels of MUC3, MUC4, and MUC12, which was accompanied by higher gene expressions of signaling kinases and transcription factors involved in mitogen-activated protein kinase (MAPK) signaling pathways. CONCLUSIONS: Based on our results, butyrate is an effective regulator of modulation of mucin protein production at the transcriptional and translational levels, resulting in changes in the adherence of gut microflora. Butyrate potentially stimulates the MAPK signaling pathway in intestinal cells, which is positively correlated with gut defense.

• 한국축산식품학회지
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• 제34권2호
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• pp.192-199
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• 2014

The effects of adding mechanically deboned chicken (MDC) hydrolysates on the quality characteristics of imitation crab stick (ICS) during storage were investigated. ICS was prepared from Alaska Pollack, chicken breast surimi, and protein hydrolysates enzymatically extracted from MDC. ICS samples were divided into 4 groups: without protein hydrolysate (control), added with 0.5% protein hydrolysate (T1), added with 1.0% protein hydrolysate (T2), and added with 1.5% protein hydrolysate (T3). Results showed that crude protein content did not differ significantly among the ICS samples (p>0.05). ICS sample added with MDC hydrolysates had higher crude fat and ash content but lower moisture content than the control (p<0.05). Lightness was significantly lower in T2 and T3 than in the other groups at 0 and 4 wk of storage. Also, whiteness decreased in the groups contained MDC hydrolysates. Breaking force and jelly strength were higher in samples containing MDC hydrolysates compared to control samples (p<0.05). Additionally, saturated fatty acid contents were lower in the groups containing MDC hydrolysates than in control sample groups (p<0.05). Polyunsaturated fatty acid (PUFA) and essential fatty acids (EFA) were significantly higher in T2 and T3 than the control samples. In particular, all samples containing MDC hydrolysates had reduced thiobarbituric acid-reactive substances (TBARS) values at 4 wk. Free radical scavenging activity also was increased with addition of MDC hydrolysates.

• Molecules and Cells
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• 제24권3호
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• pp.437-440
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• 2007

$OGL-20P^T$-358 is a novel 66 amino acid residue protein from the hyperthermophile Thermococcus thioreducens sp. nov., strain $OGL-20P^T$, which was collected from the wall of the hydrothermal black smoker in the Rainbow Vent along the mid-Atlantic ridge. This protein, which has no detectable sequence homology with proteins or domains of known function, has a calculated pI of 4.76 and a molecular mass of 8.2 kDa. We report here the backbone $^1H$, $^{15}N$, and $^{13}C$ resonance assignments of $OGL-20P^T$-358. Assignments are 97.5% (316/324) complete. Chemical shift index was used to determine the secondary structure of the protein, which appears to consist of primarily ${\alpha}$-helical regions. This work is the foundation for future studies to determine the three-dimensional solution structure of the protein.

• 정보저장시스템학회논문집
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• 제9권1호
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• pp.28-31
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• 2013

We proposed a method to fabricate label-free protein sensor with sub-wavelength nanograting structures to be used for diagnosing acute myocardial infarction. A nickel stamp for the injection molding of nanograting integrated protein sensor was fabricated by electroforming process with high fidelity. By using metallic stamp, we replicated label-free protein sensor via injection molding, which is an outstanding method for low-cost and mass production of polymer products. Finally, we performed a feasibility test, examining cardiac troponin T (cTnT) and anti-cTnT interactions. From the results, we demonstrated that the fabricated protein sensor can provide information for the early and accurate detection of cardiac diseases such as acute myocardial infarction.

• Clinical and Experimental Pediatrics
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• 제45권11호
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• pp.1368-1372
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• 2002

목 적: A군 연쇄구균의 세포벽은 M단백과 T단백으로 구성되어 있고, M단백의 항원성은 현재 90여 가지로 분류되고 있으며 균의 독성과 관련이 있고 질병의 병인론을 연구하는데 도움이 된다. 반면에 T단백은 항원성 분류는 연쇄구균의 역학적 상황을 이해하는데 유효하다고 알려져 있다. 이에 저자들은 지난 3년 동안 서울시내 5개 지역 병의원의 외래 및 입원환자를 대상으로 분리된 A군 연구균의 T단백 분류를 통해 역학적 동태를 알고자 하였다. 방 법: 1998년부터 2000년까지 서울 시내(남부, 북동부, 북부, 북서부, 북부)를 5개 지역으로 분류하여 외래 및 입원환자를 대상으로 분리된 A군 연구균의 T단백의 혈청형 분석을 시행하여 연도별 A군 연구균의 혈청학적 형 분류와 침투성질환에서 분류된 연구균의 혈청학적 형 분류의 특성을 알아보았다. 결 과 :1998년에 5개 지역에서 시행한 92례의 아동에서 A군 연구균을 분리하여 T12, T4, Non-typed(NT)가 72.2%를 차지하였고, 이중 성홍열은 7례가 있었으며 T12와 T4가 각각 3, 4례를 차지하였으며 경부임파선염 1례(T12), 편도농양 2례(T8, NT)이었다. 1999년에는 41례의 아동에서 시행하였으며 T12, T4, T1가 68%의 분포를 보였으며 성홍열은 5례 중 T12, T4가 3례를 차지하였고 폐렴 2례(T4, T1)와 경부임파선염 1례(T8/25)를 나타내었다. 2000년도는 서울 중부를 제외한 지역에서 시행한 83례의 아동에서 T12, T4, T1이 전체의 63.9%를 보였고 성홍열은 3례(T12, T4, T5 각각 1례)와 편도농양 T12 1례와 폐렴 NT 1례, 패혈증 T1 1례를 나타내었다. 결 론: A군 연구균의 T단백의 혈청학적 분석은 서울 내에서의 지역적 특정 분포는 없으나 T12가 감소하는 연도별 차이를 보였다. 성홍열이나 침투성 질환에서 특정 T단백의 분포도를 보이지는 않았다. 이에 T단백의 혈청학적 분석이 질병의 진단과 치료에 도움이 될 것으로 사료되며 앞으로 주기적으로 더 많은 아동을 대상으로 하는 역학적 검사가 이루어 져야할 것이다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권8호
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• pp.1186-1194
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• 2019

Objective: The influence of broiler carcass scalding conditions on chicken breast meat quality parameters was investigated. Methods: Two hundred and seventy Cobb broiler chickens from 42 to 48 days old were slaughtered according to the standard industry practice and scalded in five temperature/time combinations-$T_1$, $54^{\circ}C/210s$; $T_2$, $55^{\circ}C/180s$; $T_3$, $56^{\circ}C/150s$; $T_4$, $57^{\circ}C/120s$; $T_5$, $58^{\circ}C/90s$. Results: Scalding temperature increase resulted in higher values of external and ventral lightness and in protein functionality reduction-determined by emulsification capacity and protein denaturation-in chicken breast fillets 24 h post-mortem. Protein secondary structures had conformational changes, with a decrease of the ${\alpha}$-helix and an increase of the ${\beta}$-sheet and ${\beta}$-turn proportions, mainly in $T_1$ and $T_5$ samples, determined by Fourier-transform infrared spectroscopy in an attenuated reflectance mode analysis. The chemical composition, pH, water holding capacity and Warner-Bratzler shear force did not differ among the treatments. In the fatty acid profile, the 18:1n-9 was lower in $T_5$, which suggested that the high scalding-temperature could have caused the lipid oxidation. The values of the polyunsaturated fatty acids (PUFA), such as 22:2, 20:4n-6, and 22:6n-3, were highest in the $T_5$, thus being related to the phospholipid cellular membrane collapse in this experimental condition and subsequent release of these PUFA. Conclusion: Intermediate scalding-parameters avoided the negative changes in the chicken meat quality.