• Asian Pacific Journal of Cancer Prevention
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• 제14권3호
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• pp.1995-1998
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• 2013

Objective: To study the expression of the mismatch repair proteins hMSH2 and nm23 in sporadic colorectal cancer, determine any inter-relationship, and further investigate any clinical significance. Methods: Expression of hMSH2 and nm23 proteins was assessed in 87 colorectal cancer tissues by SP immunohistochemistry, with analysis of survival using follow-up data. Results: In the sporadic colorectal cancer tissues, nm23 protein expression appeared independent of the histological type (P>0.05), but correlated with the invasion depth and lymphatic metastasis (P<0.05). In contrast, hMSH2 protein expression was not significantly correlated with these clinicopathologic features (P>0.05), although it positively correlated with that of nm23 protein in the sporadic colorectal cancers (rs=0.635, P<0.05). Combined expression of the two was found to be related with invasion depth, lymphatic metastasis and prognosis of sporadic colorectal cancer (P<0.05). Conclusion: nm23 protein level was related with the degree of malignancy, and could be used as an index to predict the invasion and metastasis potential. The expression of hMSH2 protein is positively correlated that of nm23 protein, and the combined expression of the two has certain guiding significance for the prognosis of sporadic colorectal cancer.

• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.507-513
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• 2017

Bacillus subtilis spores can be used for protein display to engineer protein properties. This method overcomes viability and protein-folding concerns associated with traditional protein display methods. Spores remain viable under extreme conditions and the genotype/phenotype connection remains intact. In addition, the natural sporulation process eliminates protein-folding concerns that are coupled to the target protein traveling through cell membranes. Furthermore, ATP-dependent chaperones are present to assist in protein folding. CotA was optimized as a whole-cell biocatalyst immobilized in an inert matrix of the spore. In general, proteins that are immobilized have advantages in biocatalysis. For example, the protein can be easily removed from the reaction and it is more stable. The aim is to improve the pH stability using spore display. The maximum activity of CotA is between pH 4 and 5 for the substrate ABTS (ABTS = diammonium 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate). However, the activity dramatically decreases at pH 4. The activity is not significantly altered at pH 5. A library of approximately 3,000 clones was screened. A E498G variant was identified to have a half-life of inactivation ($t_{1/2}$) at pH 4 that was 24.8 times greater compared with wt-CotA. In a previous investigation, a CotA library was screened for organic solvent resistance and a T480A mutant was found. Consequently, T480A/E498G-CotA was constructed and the $t_{1/2}$ was 62.1 times greater than wt-CotA. Finally, E498G-CotA and T480A/E498G-CotA yielded 3.7- and 5.3-fold more product than did wt-CotA after recycling the biocatalyst seven times over 42 h.

• Applied Biological Chemistry
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• 제36권2호
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• pp.121-126
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• 1993

참깨박에 함유되어 있는 불용성 형태의 단백질을 가용성 형태의 단백질로 용출시키기 위하여 Bacillus sp. CW-1121이 생성하는 효소를 작용시켰다. 이때 참깨박의 단백질 용출을 위한 최적 pH는 7.5였으며, 최적 온도는 $40^{\circ}C$였고 최적 조건하에서 2시간의 소화로 약 60%의 용해도를 보였다. 단백질의 용해도는 참깨 단백질이 pH4.5에서 가장 낮아 등전점을 보였고, pH3.0과 pH6.0 이상에서는 크게 증가하였다. 효소처리 참깨박 단백질을 전기영동한 결과 수용성 단백질은 4개, 염용성 단백질은 2개의 밴드가 관찰되었다. 분리 단백의 아미노산 조성은 수용성 단백질의 경우 serine이 17.24 mg/g, 염용성 단백질은 glutamic acid가 10.77 mg/g, 유리아미노산은 glutamic acid가 6.55 mg/g으로 가장 많이 함유되어 있었으며 특히 필수아미노산의 조성도 상당히 좋은 편이었다.

• 농업과학연구
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• 제43권3호
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• pp.379-386
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• 2016

Four ruminally cannulated Holstein steers (BW $482.9{\pm}8.10kg$), fed low protein TMR (CP 11.7%) as a basal diet, were used to investigate changes in rumen fermentation and blood metabolism according to protein fraction, cornell net carbohydrates and protein system (CNCPS), and enriched feeds. The steers, arranged in a $4{\times}4$ Latin square design, consumed TMR only (control), TMR supplemented with rapeseed meal (AB1), soybean meal (B2), and perilla meal (B3C), respectively. The protein feeds were substituted for 23.0% of CP in TMR. Ruminal pH, ammonia-N, and volatile fatty acids (VFA) in rumen digesta, sampled through ruminal cannula at 1 h-interval after the morning feeding, were analyzed. For plasma metabolites analysis, blood was sampled via the jugular vein after the rumen digesta sampling. Different N fraction-enriched protein feeds did not affect (p > 0.05) mean ruminal pH except AB1 being numerically lower 1 - 3 h post-feeding than the other groups. Mean ammonia-N was statistically (p < 0.05) higher for AB1 than for the other groups, but VFA did not differ among the groups. Blood urea nitrogen was statistically (p < 0.05) higher for B2 than for the other groups, which was rather unclear due to relatively low ruminal ammonia-N. This indicates that additional studies on relationships between dietary N fractions and ruminant metabolism according to different levels of CP in a basal diet should be required.

• 한국식품영양과학회지
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• 제35권10호
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• pp.1394-1398
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• 2006

pH 전이공정에 따라 어육 단백질을 회수하고 드럼건조하여 분말 단백질을 제조한 후, 이들 단백질의 몇 가지 식품학적 기능성을 측정하였다. 건조 어육 단백질 분말의 용해도는 pH 7.0으로 조절한 단백질이 PH 5.5에 비하여 높았다. 유화활성지표는 $0.035\sim0.043m^2/g-protein$(pH 5.5 단백질 분말)로 비교적 낮았다. 그리고 유화안정성, 거품능 및 거품안정성은 관측되지 않았다. 점도 값은 수리미 단백질의 1/10에 불과한 $50,200\sim39,000\;cP$의 범위였다. 수분과 지방흡착능력은 각각 $2.63\sim2.89g-water/g$과 $2.13\sim2.17g-oil/g$이었고, 입자의 크기와 pH에 따른 영향은 없었다. 이 같은 실험 결과는 지방흡착을 위한 육의 대체제로 patty 제품 등에 건조 어육 단백질을 적용할 수 있을 것으로 보인다.

• 대한의생명과학회지
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• 제2권2호
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• pp.175-185
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• 1996

모든 진핵세포에 존재하며 세포의 성장 및 분화에 주로 관계되는 신호전달물질의 하나인 Mitogen-activated protein(MAP)kinase의 mitogen에 의한 핵내 활성화와 기질 인산화에 대해 알아보기 위해 본 실험을 수행하였다. P388세포를 10% fetal bovine serum이 첨가된 DMEM배지에 배양한 후, 혈청이 들어있지 않은 배지에서 24시간 더 배양하고 serum 및 PMA를 농도별로 처리하여 세포성 장을 위한 최적 농도를 확인한 결과 serum은 5-20% 농도에서 세포성장을 촉진시켰고 PMA는 실험한 모든 농도에서 세포성장을 거의 촉진시키지 못하는 경향을 확인하였다. 이어 P388 세포를 serum 및 PMA로 10 분간 활성화하여 파쇄한후 세포질분획과 핵분획으로 분리하여 각 분획을 10% gel 상에서 전기영동 하여 nitrocellulose paper에 옳긴 후 anti-ERKI antibody를 이용해 확인해본 결과 serum, PMA로 처리된 세포 모두에서 MAP kinase의 핵내 이동이 관찰되었으며 특히 세포질 내에 주로 존재하는 42, 44 Kd의 MAP kinase isoform중 42 Kd의 isoform이 주로 핵내로 이동되는 것이 관찰되었다. MAP kinase의 기질인산화 실험을 위해 serum으로 활성화시킨 세포를 파쇄하여 SP-sephadex C-50, Phenyl superose, Mono Q column의 순서로 chromatography를 시 행하여 MAP kinase를 부분분리 하였다. 이와 같이 얻은 MAP kinase를 가지고 면역 T세포에 존재하는 tyrosine kinase인 $p56^{lck}$ 의 N-terminal peptide로 구성된 GST-fusion protein에 대한 인산화를 확인하였다. 또한 세포에서 분리한 MAP kinase를 가지고 transcription factor의 하나인 c-Jun protein에 대한 인산화실험을 실시한 결과 MAP kinase에 의해 인산화 됨이 확인되었다. 이상의 결과를 통해 P388세포는 (1)세포 성장시 외부 신호를 G-protein-coupled receptor/protein kinase C/MAP kinase의 경로보다는 주로 tyrosine kinase receptor protein/Ras/MAP kinase의 경로를 이용하여 핵으로 전달하는 것으로 추측되 며 (2) mitogen의 처리로 활성화된 MAP kinase중 주로 42 Kd isoform이 핵내로 이동하고, 분리한 MAP kinase가 GST-fusion protein과 transcription factor인 c-Jun을 모두 인산화 시키는 결과로 보아 MAP kinase의 isoform에 따라 표적 compartment가 다르고 결과적으로 표적 기질에 차이가 있을지 모른다고 간접적으로 추론할 수 있다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2022년도 추계학술대회
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• pp.104-104
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• 2022

Dendropanax morbiferus H. Lev has been reported to have some pharmacologic activities and also interested in functional cosmetics. We found that the water extract of D. morbiferus leaves significantly inhibited tyrosinase activity and melanin formation in α-melanocyte stimulating hormone (MSH)-induced B16-F10 cells. D. morbiferus reduced melanogenesis-related protein levels, such as microphthalmia? associated transcription factor (MITF), TRP-1, and TRP-2, without any cytotoxicity. Two active ingredients of D. morbiferus, (10E)-9,16-dihydroxyoctadeca-10,17-dien-12,14-diynoate (DMW-1) and (10E)-(?)-10,17-octadecadiene-12,14-diyne-1,9,16-triol (DMW-2) were identified by testing the anti-melanogenic effects and then by liquid chromatography-tandem mass spectrometry (LC/MS/MS) analysis. DMW-1 and DMW-2 significantly inhibited melanogenesis by the suppression of protein kinase A (PKA)/cyclic AMP (cAMP)-responsive binding protein (CREB) and p38 MAPK phosphorylation. DMW-1 showed a better inhibitory effect than DMW-2 in α-MSH-induced B16-F10 cells. D. morbiferus and its active component DMW-1 inhibited melanogenesis through the downregulation of cAMP, p-PKA/CREB, p-p38, MITF, TRP-1, TRP-2, and tyrosinase. These results indicate that D. morbiferus and DMW-1 may be useful ingredients for cosmetics and therapeutic agents for skin hyperpigmentation disorders.

• Asian-Australasian Journal of Animal Sciences
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• 제23권3호
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• pp.333-339
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• 2010

The objectives of this study were to use the NRC-2001 model and DVE/OEB system to model potential nutrient supply to ruminants and to compare frost damaged (also called "frozen" wheat with normal wheat. Quantitative predictions were made in terms of: i) Truly absorbed rumen synthesized microbial protein in the small intestine; ii) Truly absorbed rumen undegraded feed protein in the small intestine; iii) Endogenous protein in the digestive tract; iv). Total truly absorbed protein in the small intestine; and v). Protein degraded balance. The overall yield losses of the frozen wheat were 24%. Results showed that using the DVE/OEB system to predict the potential nutrient supply, the frozen wheat had similar truly absorbed rumen synthesized microbial protein (65 vs. 66 g/kg DM; p>0.05), tended to have lower truly absorbed rumen undegraded feed protein (39 vs. 53 g/kg DM; p<0.10) and had higher endogenous protein (14 vs. 9 g/kg DM; p<0.05). Total truly absorbed protein in the small intestine was significantly lower (89 vs. 110 g/kg DM, p<0.05) in the frozen wheat. The protein degraded balance was similar and both were negative (-2 vs. -1 g/kg DM). Using the NRC-2001 model to predict the potential nutrient supply, the frozen wheat also had similar truly absorbed rumen synthesized microbial protein (average 56 g/kg DM; p>0.05), tended to have lower truly absorbed rumen undegraded feed protein (35 vs. 48, g/kg DM; p<0.10) and had similar endogenous protein (average 4 g/kg DM; p>0.05). Total truly absorbed protein in the small intestine was significantly lower (95 vs. 108 g/kg DM, p<0.05) in the frozen wheat. The protein degraded balance was not significantly different and both were negative (-16 vs. -19 g/kg DM). In conclusion, both models predict lower protein value and negative protein degraded balance in the frozen wheat. The frost damage to the wheat reduced nutrient content and availability and thus reduced nutrient supply to ruminants by around 12 to 19%.

• Applied Biological Chemistry
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• 제13권1호
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• pp.81-85
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• 1970

한국답토양십종(韓國畓土壤十種)을 시료(試料)로하여 유효인산정량(有動燐酸定量)에 적합한 화학적(化學的) 추출방법(油出方法)을 찾고저 A-value인(燐)-32 추적자법(追跡子淪)에 의한 유효인산정량치(有效燐酸定量値)를 표준으로 하여 시험한 결과 다음과 같은 결과(讀果)를 얻었다. 1 A·value는 Fe-P(인산철(燐酸鐵))와만 1%수준(水準)의 유의상관(有意相鬪)이 있으므로 답토양(畓土壤)의 유효인산(有刻燐酸)은 Fe·p계(系)에 의하여 지배(支配)되고 있다고 볼 수 있다. 2. 본(本) 실험(實驗)에 사용(使用)한 6개침출법(個浸出法)은 Fe-P와 상안(相關)이 없으므로 답토양유효인산정량법(容土壤有效燐酸定量法)으로 적합하지 않다. 3. 답토양유효인산(容土壤有效燐酸)에 적합한 침출법(浸出法)은 Fe-P계(系)의 환원강도(還元强度)를 측정(測定)할 수 있는 것이어야 하며 침출강도(浸出强度)는 평균(平均) 270 ppm 으로 추정(推定)된다. 4. Bray No. 2-p는 Lancaster-p, Spurway-p, Truog-p 및 (Ca+Al)-p와 5% 수준(水準) 이상(以上)에서 유의상관(有意相關)이 있고 Al-P는 Olsen-P 및 Lancaster-p와, Ca-p는 Spurway-p와 1% 수준(水準)에서 유의상관(有意相關)이 있다.

• 대한영양사협회학술지
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• 제13권4호
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• pp.301-310
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• 2007

The purpose of this study was to analyze the relationships among zinc status, protein and phytate intake, and diabetic control indices of type 2 diabetic women. The mean age and the duration of diabetes were respectively 57.9±6.9 years old and 8.0±6.5 years. The mean daily energy intake of diabetic subjects was 1562 kcal. Both the zinc intake (6.2mg/day) and the zinc %RI (% of The Recommended Intake for zinc: 79.5%) of the diabetic participants were significantly lower than those of the control group (respectively p<0.01). As for the diabetic group, the higher the energy intake (kcal/day), the higher were the zinc intake (p<0.001) and %RI for zinc (p<0.001). Zinc intake was positively correlated with the protein (p<0.001), animal protein (p<0.001), and fat intake (p<0.001), but negatively correlated with the carbohydrate intake (p<0.001). Foods with high amount of phytate were the major source of zinc (p<0.01), but did not contribute to high zinc densities. The urinary zinc excretion was twice as high as in the diabetic group compared to the control group (p<0.001). In addition, the urinary zinc loss was positively correlated with the duration of diabetes (p<0.05), hyperglycemia (p<0.001) and insulin resistance (p<0.05). %RI for zinc was negatively correlated with the HbA1C (p<0.05). These results lead us to conclude that the appropriate intake of energy controlled by diet therapy could improve the total zinc intake and %RI for zinc in diabetic women. Also, normal blood glucose level controlled by diet therapy could improve the hyperzincuria. Dietetic practitioners should encourage consumption patterns that provide zinc-rich foods in the form of animal protein to improve the bioavailability as well as the total daily intake of zinc.