• Animal Bioscience
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• 제36권7호
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• pp.1120-1129
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• 2023

Objective: This study aimed to determine the protective efficacy of Buddha's Temple (BT) extract against tert-butyl hydroperoxide (t-BHP)-induced oxidative stress in Gallus gallus chicken embryo fibroblast cell line (DF-1) and its effects on the cell lipid metabolism. Methods: In this experimental study, Gallus gallus DF-1 fibroblast cells were pretreated with BT 10^-7 for 24 hours, followed by their six-hour exposure to t-BHP (100 μM). Water-soluble tetrazolium salt-8 (WST-8) assays were performed, and the growth curve was computed. The intracellular gene expression changes caused by BT extract were confirmed through quantitative polymerase chain reaction (qPCR). Flow cytometry, oil red O staining experiment, and thin-layer chromatography were performed for the detection of intracellular metabolic mechanism changes. Results: The WST-8 assay results showed that the BT pretreatment of Gallus gallus DF-1 fibroblast cell increased their cell survival rate by 1.08%±0.04%, decreased the reactive oxygen species (ROS) level by 0.93%±0.12% even after exposure to oxidants, and stabilized mitochondrial activity by 1.37%±0.36%. In addition, qPCR results confirmed that the gene expression levels of tumor necrosis factor α (TNFα), TIR domain-containing adapter inducing IFN-beta (TICAM1), and glucose-regulated protein 78 (GRP78) were regulated, which contributed to cell stabilization. Thin-layer chromatography and oil red O analyses showed a clear decrease in the contents of lipid metabolites such as triacylglycerol and free fatty acids. Conclusion: In this study, we confirmed that the examined BT extract exerted selective protective effects on Gallus gallus DF-1 fibroblast cells against cell damage caused by t-BHP, which is a strong oxidative inducer. Furthermore, we established that this extract significantly reduced the intracellular ROS accumulation due to oxidative stress, which contributes to an increase in poultry production and higher incomes.

• 한국가축번식학회지
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• 제27권1호
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• pp.9-14
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• 2003

형질전환 동물의 유선에서 특이적으로 인간 TPO가 발현되도록 고안된 pBT-L 외래유전자가 삽입되어 한 계통으로 확립된 형질전환 생쥐에서 이 유전자가 장기 세대까지 안정적으로 전이되고, 또한 발현 수준도 지속적으로 유지되는지를 조사 하였다 이를 위해 제 5세대의 수컷 pBT-L 형질전환 생쥐를 실험에 공시하였고, 제 10세대까지의 전이율과 인간 TPO의 발현수준을 분석하였다. 제 8세대 생쥐의 계대 번식에 의한 자손 중 51.3$\pm$18.98%가 형질전환 생쥐로 판명되었다. 또한 제9세대에서 외래 유전자의 전이율은 43.8$\pm$18.98%이었고, 제 10세대에서는 71.4$\pm$26.98%의 전이율을 나타내었다. 이러한 결과로 pBT-L 형질전환 생쥐는 그 외래유전자의 유전적 손상이 없이 장기 세대까지 안정적으로 전이되는 것으로 판단된다. 제 9세대외 10세대의 3마리의 암컷 형질전환 생쥐로 부터 유즙내 인간 TPO의 발현 수준을 분석하였을때, 그 농도는 모두 평균 1.1 mg/ml의 수준에서 측정되었다. 이러한 수준은 제 2세대의 것과 유사한 결과로 pBT-L 형질전환 생쥐에서 인간 TPO 유전자의 발현은 최소한 10세대까지 지속적으로 유지된다는 것을 알 수 있었고, 더 긴 장기 세대까지도 발현 수준이 유지될 것으로 추정된다. 이러한 연구결과는 계통으로 확립된 형질전환동물에 부여된 새로운 유전형질은 장기 세대까지 유전될 수 있음을 보여주는 것이다.

• 한국수산과학회지
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• 제43권5호
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• pp.445-450
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• 2010

This study investigated the effects of dietary lipid sources on growth performance and body composition of juvenile far eastern catfish, Silurus asotus. Three replicate groups of fish (average weight 3.6 g) were fed with one of the following experimental diets containing 10% beef tallow (BT), 5% BT plus 5% corn oil (CO), 5% BT plus 5% linseed oil (LO), or 5% BT plus 5% squid liver oil (SO) as the lipid source for 5 weeks. No significant difference was observed in the survival among groups. The weight gain of fish fed the LO (high in 18:3n-3) and SO (high in n-3 highly unsaturated fatty acid) diets was significantly higher than that of the fish fed the CO (high in 18:2n-6) and BT diets (P<0.05). The feed efficiency of fish fed LO and SO diets was significantly higher than that of the fish fed the BT diet (P<0.05), but not significantly different from that of the fish fed the CO diet. The protein efficiency ratio of fish fed the SO diet was significantly higher than that of fish fed the CO and BT diets (P<0.05), but not significantly different from that of fish fed the LO diet. The 18:1n-9 of whole-body polar lipid fraction in fish fed the BT diet increased compared to that of fish fed the other diets. Fish fed the CO and LO diets had significantly higher contents of 18:2n-6 and 20:4n-6, and 18:3n-3, than the fish fed the other diets in polar and non-polar lipid fractions, respectively (P<0.05). Significantly higher contents of 20:5n-3 and 22:6n-3 were observed in the whole-body polar lipid fraction of fish fed the SO diet compared with fish fed the other diets (P<0.05). The study results indicate that linseed oil and squid liver oil containing n-3 fatty acids are good dietary lipid sources for the growth of far eastern catfish.

• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.1093-1098
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• 2001

The gene coding for a Lepidoptera-specific insecticidal crystalline (or control) protein (ICP), recognized as cryIAc, from Bacillus thuringiensis subsp. kurstaki HD-73, was cloned into the vector pBluscript ll SK-, and then transformed in Escherichia coli $DH5{\alpha}$. The clone was named EBtIAc and the chimeric phagemid, as pEBtIAc. Hyperexpression of CryIAc protoxin was observed in the extract of the culture of E. coli harboring pEBtIAc. Crystalline protoxin was purified by differential solubility. It was dissolved in alkaline pH, and exposed to trypsin to be activated. The molecular weights of the pro- and activated toxins on SDS-PAGE were estimated to be ca. 130 kDa and 60 kDa, respectively. The toxicity was tested by force-feeding larvae of gypsi moth (Lymantria diapar) with trypsinized protoxin. Using the batch of biologically active form of the toxin as an immunogen, anti-CryIAc antiserum was raised in a New Zealand white rabbit. Immunoglobulin G was fractionated from the seam by Protein-A sepharose affinity chromatography. Immunoreactivity of the antibody was examined by dot and Westerns blottings. It has been found that the anti- CryIAc antibody recognized the purified toxin at a level below a nanogram in terms of quantity. Using the antibody some of Bt-corns were able to be differentiated from tons of corn kernels which were imported from America as forage crops.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3265-3270
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• 2012

Clinically, elevated cancer antigen 125 (CA-125) in blood predicts tumor burden in a woman's body, especially in the ovary, but cannot differentiate between malignant or benign. We here used intensive modern proteomic approaches to identify predictive proteins in the serum of women with elevated CA-125 to differentiate malignant from benign ovarian tumors. We identified differentially expressed proteins in serum samples of ovarian cancer (OC) patients, benign ovarian tumor (BT) patients, and healthy control women using mass spectrometry-based quantitative proteomics. Both the OC and BT patients had elevated CA-125. Quantitation was achieved using isobaric tags for relative and absolute quantitation. We obtained 124 quantified differential serum proteins in OC compared with BT. Two proteins, apolipoprotein A-4 (APOA4) and natural resistance-associated macrophage 1, were verified using Western blotting. Proteome profiling applied to OC cases identified several differential serum proteins in the serum of women with elevated CA-125. A novel protein, APOA4, has the potential to be a marker for malignant tumor differentiation in the serum of women with elevated CA-125.

• 한국잠사곤충학회지
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• 제36권2호
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• pp.162-167
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• 1994

Bacillus thuringiensis var. kurstaki의 내독소단백질을 섭취한 Helocoverpa assulta의 유충에서 각 조직의 표면구조변화를 주사전자현미경으로 조사하였다. Bt 내독소단백질은 조사된 모든 조직 표면에서 부식과 파괴현상을 유발시켰다. 이러한 결과들로 볼 때 독소분자들은 세포막에 대한 뚜렷한 특이성이 없이 독소에 노출된 모둔 종류의 세포막에 결합하는 것으로 보인다.

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2003년도 International Symposium of Silkworm/Insect Biotechnology and Annual Meeting of Korea Society of Sericultural Science
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• pp.118-119
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• 2003

Baculovirus occlusion bodies have been recently engineered to incorporate foreign protein such as the Bacillus thuringiensis (Bt) CrylAc protein for improvement of insecticidal activity. In this study, polyhedrin, cylAc, egfp and crylCa genes were fused to produce occlusion bodies that contain novel recombinant crystal protein by homologous recombination between cylAc and crylCa genes in insect cells. (omitted)

• 셀메드
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• 제3권2호
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• pp.18.1-18.7
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• 2013

Inflammation in rheumatoid arthritis is characterized by immune cell infiltration and cytokine secretion. In particular, mast cells and their cytokines play an important role in the pathogenesis of rheumatoid arthritis. Korean medicine, BaekJeol-Tang (BT) was designed by traditional Korean medicine theory. We already reported therapeutic effect of BT in rheumatoid arthritis. Here, we report the specific underlying mechanism of BT in activated human mast cells, HMC-1 cells. In addition, we report for the first time that BT significantly inhibited the production and mRNA expression of proinflammatory cytokines including thymic stromal lymphopoietin, interleukin (IL)-$1{\beta}$, IL-6, IL-8, and tumor necrosis factor-${\alpha}$ in activated HMC-1 cells. BT also decreased the activation of mitogen-activated protein kinases, nuclear factor-${\kappa}B$, and caspapase-1. Taken together, these results indicate that BT has potential as a regulator of inflammatory reactions for the treatment of arthritis such as osteoarthritis and rheumatoid arthritis.

• 한국가금학회지
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• 제49권4호
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• pp.273-279
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• 2022

The objective of this study was to evaluate an in vitro procedure to estimate the crude protein (CP) digestibility of feed ingredients and mixed diets in broiler chickens. The apparent ileal digestibility (AID) of the CP was measured using 23-d-old male broilers. Three experimental diets, containing three feed ingredients, namely soybean meal (SBM), canola meal (CM), and corn distiller's dried grains with solubles (DDGS), were used as the sole source of CP. A 2-step in vitro procedure was used to estimate in vivo CP digestibility; all the experiments were performed in triplicate. In step 1, the feed ingredient and mixed diet samples were incubated for 4 h at 40 °C with a pH 2.0 pepsin solution, and in step 2, the flasks were incubated for 12 h at 40 °C with a pH 6.8 pancreatin solution. Following incubation, all the samples were filtered; the undigested residues were collected and pooled together to analyze the undigested CP concentration. The in vitro CP digestibility of mixed diets and feed ingredients were 93.2% and 93.0% for SBM, 86.8% and 86.7% for CM, and 83.8% and 79.1% for DDGS, respectively. The coefficients of determination (R2) between the CP digestibility values for the feed ingredients and the in vitro CP digestibility values for the feed ingredients or respective mixed diets were 0.87 or 0.67. The results of the study demonstrated that the in vitro CP digestibility values obtained from the respective mixed diets were better estimates than the values obtained from the individual feed ingredients to predict the AID of CP in feed ingredients fed to broiler chickens.

• 한국미생물·생명공학회지
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• 제17권4호
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• pp.295-300
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• 1989

B. thuringiensis가 생산하는 살충성 독소 단백질의 생태학적 응용방법을 개발하기 위한 목적으로 우선 독소 단백질 유전자를 옮겨 발현시키기에 적합한 숙주 미생물의 분리작업을 수행하였다. 국내 주요농산물인 고추, 감자, 무우 등 7가지 농작물의 뿌리부근에 군락을 형성하는 35종의 형광성Pseudomonas들을 분리하였고 독소 단백질 유전자를 함유하는 재조합 plasmid에 대한 숙주로서의 이응가능성을 검토해 보기 위하여 분리균주 35주에 대한 형질전환을 실시한 결과 4주에 독소 단백질 유전자의 도입이 가능하였고 생물검정과 면역학적인 방법 등에 의한 결과 BT 독소 유전자의 발현을 확인하였다.