• Title/Summary/Keyword: p21 단백

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Enzymatic Properties of Cytidine Deaminase Encoded by cdd Gene in Bacillus subtilis (Bacillus subtilis의 cdd 유전자에 의해 코드되는 Cytidine Deaminase의 효소학적 성질)

  • Song, Bang-Ho;Yoon, Mi-Sook;Kim, Kyung-Hwa;Yeo, Jeung-Sook;Jan Neuhard
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.468-475
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    • 1988
  • The cloned B. subtilis cdd gene encoding cytidine/2'deoxycytidine deaminase (EC 3.5.4.5) was expressed in the cdd deficient B. subtilis mutant ED40. The gene was isolted from the cdd complementing plasmid pSO21, and inserted into the EcoR1/Pvu1 sites of pGB215-110 ΔB, which is a temperature sensitivie E. coli-B. subtilis shuttle vector. In the transformed B. subtilis ED4O harboring the resulting plasmid pSO100, cdd was expressed at several hundred fold elevated levels, and the cytidine deaminase activity in E. coli containing pSO100 was twice the level in B. subtilis/pSO0100. The Km value for cytidine of the partially purified enzyme is 1.88$\times$10$^{-4}$M at pH 7.0 and the V$_{max}$ = 11.1 $\mu$mol/min/mg of protein. The enzyme was completely inhibited by 0.1M mercaptoethanol and HgCl$_2$. The inhibition by p-chrolomercurybenzoic acid showed a Ki = 5 uM. These results suggest that sulfhydryl reagents block an active site thiol group, and/or disturb the formation of the tetrameic holoenzyme.

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Production of Plant Protein Concentrate and Yeast Biomass from Radish Greens (무청즙액을 이용한 녹엽단백질과 효모균체의 생산)

  • Rhee, Yeong-Sang;Kyung, Kyu-Hang;Yoo, Yang-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.3
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    • pp.263-269
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    • 1992
  • Radish green juice was used as a dual source for the production of plant protein precipitate and Candida utilis biomass. Precipitates ranging from 10.0 to 16.5g were obtained from a liter of radish green juice by heating at 80-10$0^{\circ}C$C for 1 to 10 min or by modification of the pH of radish green juice. Crude protein content of the precipitate was between 25 and 38%. The residue remaining after protein precipitation was used in turn for the cultivation of the yeast, C. utilis, in order to produce yeast biomass. C. utilis grew well in radish green residual juice and completed growth within 24 hr at 3$0^{\circ}C$ and 200rpm in shake flask experiments. Maximum dry cell weight obtainable from a liter of radish green residual juice was 19.5g, when the yeast was grown on the juice residue diluted 3 times or more with water to make sugar content be equal to or less than about 1.0%. Supplementation of 3-fold diluted radish green residual juice with yeast extract and (NH$_4$)SO$_4$ enhanced yeast biomass production and cell protein content significantly. Total high protein material obtainable from a liter of radish green juice was 33.0g.

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Cloning and protein expression of Actinobacillus actinomycetemcomitans cytolethal distending toxin subunit CdtA (Actinobacillus actinomycetemcomitans의 cytolethal distending toxin subunit CdtA 유전자 클로닝과 단백질 발현)

  • Ko, Sun-Young;Jeong, Dong-Keun;Ryu, So-Hyun;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.37 no.sup2
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    • pp.339-351
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    • 2007
  • Cytolethal distending toxin(CDT)은 세포 주기 중 G2에서 M 기로의 전환을 막아 세포의 증식을 억제할 수 있는 세균 단백 독소의 일종이다. 구강 미생물 중 유일하게 Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans)만이 이 CDT를 생성 할 수 있는 것으로 알려져 있다. A. actinomycetemcomitans는 localized aggressive periodontitis (LAP)의 원인균으로 여겨지며 비 운동성의 그람 음성 구간균이고 $37^{\circ}C$, 5% $CO_2$ 하에 성장이 왕성하다. A. actinomycetemcomitans의 CDT는 3개의 인접한 유전자인 cdtA, cdtB, cdtC에 의해 형성 되며 각각의 유전자에 대한 단백질의 기능은 아직 완전히 밝혀지지 않았다. 현재까지 연구에 의하면 cdtA는 CDT의 세포부착과 관련이 있는 것으로 여겨지며 이 유전자의 기능 이상 시 CDT의 독성 효과가 현저히 감소한다고 알려져 있다. 따라서 본 연구는 A. actinomycetemcomitans의 cdtA 유전자를 클로닝, 단백질 발현하여 향후 치주질환의 발병 과정에서 CdtA의 역할을 규명하고 질환의 예방 및 치료법에 도움을 주고자 하였다. A. actinomycetemcomitans Y4균주를 cdtA 유전자 클로닝을 위해 사용하였다. A. actinomycetemcomitans의 genomic DNA는 genomic DNA 추출 kit를 사용하여 분리하고 cdtA에 특이적인 primer를 이용하여 PCR을 통해 cdtA 유전자를 증폭하였다. 증폭된 cdtA 유전자를 T-vector에 클로닝 하였으며, 클로닝 된 cdtA 유전자는 단백질 발현을 위해 pRSET Avector에 서브클로닝 한 후 발현 균주인 BL21(DE3)를 이용하여 발현시켰다. 발현 후 Ni-NTA AP conjugate를 이용한 Western blot을 통해 pRSET-CDTA를 확인하였다.

Functionality Changes of Rapeseed Protein upon Proteolysis (유채단백질의 Proteolysis에 의한 기능성 변화)

  • Kim, Chung-Hee;Kim, Hyo-Sun;Lee, Jang-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.5
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    • pp.519-524
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    • 1992
  • purified rapeseed(Brassica napus var. Youngsan) protein was hydrolyzed by pronase. The hyrolysate protein was investigated for the some physicochemical and functional properties. UV and intrinsic fluorescence spectra of the hydrolysate showed the maximum absorption at 274nm and 360nm respectively. Intensity of yellow color decreased in the process of hydrolysis and the surface hydrophobicity decreased up to fourfold. The main bands of hydrolysate by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) were observed at 14,000 to 12,000 dalton molecular weight. Solubilities of hydrolyzed protein increased by 10~15% compared to those of unhydrolyzed protein at acidic pH. In the hydrolysate, while absorption of both water and oil, foam expansion and emulsion stability were increased, absolute viscosity, heat coagulation, calcium coagulation, foam stability and emulsion activity were decreased.

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Acid Production by Lactic Acid Bacteria in Soy Milk Treated by Microbial Pretense or Papain and Preparation of Soy Yogurt (미생물 Protease 또는 Papain으로 처리된 두유에서 젖산균의 산생함과 대두요구르트의 제조)

  • Ko, Young-Tae
    • Korean Journal of Food Science and Technology
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    • v.21 no.3
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    • pp.379-386
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    • 1989
  • The soy milk prepared from soy protein concentrate was treated with microbial protease or papain. Growth and acid production by Lactobacillus acidophilus in soy milk containing partially hydrolyzed proteins were investigated. Sensory evaluation of yogurt beverage prepared from protease treated soy milk was also performed. Protease treatment of soy milk enhanced acid production by lactic acid bacteria, particularly in case of microbial pretense and simultaneous treatment by two types of protease showed synergistic effect. pH and number of viable cells were not affected markedly by pretense treatment. Microbial pretense treatment up to 15 minutes or papain treatment up to 45 minutes enhanced acid production, but further treatment up to three hours did not affect the acidity markedly. rho sensory evaluation showed that overall acceptability and taste of soy yogurt beverage were slightly improved when soy milk was treated with microbial pretense of 0.2% or papain of 0.2%. The amount of non-protein nitrogen considerably increased by pretense treatment of 15 minutes and it increased gradually by further treatment up to three hours.

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Effects of Pretense Treatment of Soy Milk on Acid Production by Lactic Acid Bacteria and Quality of Soy Yogurt (두유(豆乳)의 단백질분해효소 처리가 젖산균의 산생성(酸生成)과 대두요구르트의 품질에 미치는 영향)

  • Kim, Kyung-Hee;Bang, Il-Ryung;Ko, Young-Tae
    • Korean Journal of Food Science and Technology
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    • v.21 no.1
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    • pp.92-99
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    • 1989
  • The soy milk prepared from soy protein concentrate was treated with pretense of various concentrations. Growth and acid production by Lactobacillus acidophilus or L. bulgaricus in soy milk containing partially hydrolyzed protein were investigated. Sensory evaluation of soy yogurt beverages prepared from pretense treated soy milk was also performed. The treatment of soy milk with pretense markedly enhanced acid production by lactic acid bacteria, particularly by L. acidophilus, whereas pH and number of viable cells were not .affected by pretense treatment. Protease treatment of 15 minutes greatly enhanced acid production by lactic acid bacteria, but further treatment up to three hours did not affect the acidity markedly. The sensory evaluation showed that overall acceptability of soy yogurt beverages was slightly improved when soy milk was treated with pretense of 0.1%. The amount of non-protein nitrogen was considerably increased by pretense treatment of 15 minutes and it reached the maximum value by treatment of two hours.

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Risk Factors for the First-Year Relapse in Children with Nephrotic Syndrome (신증후군 환아에서 1년내 재발과 관련된 위험 요인)

  • Shin, Hye Kyoung;Kim, Ji Hee;Yoo, Kee Hwan;Hong, Young Sook;Lee, Joo Won;Kim, Soon Kyum
    • Clinical and Experimental Pediatrics
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    • v.46 no.9
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    • pp.889-892
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    • 2003
  • Purpose : This study aimed to evaluate risk factors of the first year relapse in children with nephrotic syndrome(NS) without the need for biopsy. Methods : We reviewed, retrospectively, 78 children diagnosed with steroid responsive nephrotic syndrome between July 1997 and June 2002. Median years to follow up were 4.4 years(range : 1-5 years). We divided the patients into two groups(group I : primary responders with no relapse or with only two relapses in the first year after initial response; group II : initial steroid responders with three or more relapses within the first year). We retrospectively reviewed and compared variables - sex, onset age, serum albumin, serum cholesterol, 24 hours urinary protein, creatinine clearance(Ccr), presence of hematuria and hypertension(HTN), and days from initial attack to remission. Results : Of 78 patients(male : 61(78.2%), female : 17(21.8%), age range 1.1 years to 14 years, median $5.1{\pm}3.0years$), 47(60.3%) were in group I and 31(39.7%) were in group II. There were no statistically significant differences in sex, serum albumin, serum cholesterol, 24 hours urinary protein, Ccr, presence of hematuria or HTN. The median age of onset showed no statistical difference between the two groups. However, if the patients are dividing into two groups according to the age of onset of three-years, patients theree yrs old or less fit into group II, as opposed to patients older than three yrs in age(63.2% vs. 32.2%, P<0.05). The days from initial attack to remission was longer in group II($12.9{\pm}0.5$ vs. $16.2{\pm}1.1$, P<0.05). Conclusion : We may conclude that the age of onset of three yrs old or less, and the longer time remission to initial steroid therapy, are risk factors of the first year relapse.

The effect of silk fibroin and rhBMP-2 on bone regeneration in rat calvarial defect model (백서 두개골결손모델에서 실크단백과 골형성단백 이식체가 골재생에 미치는 영향)

  • Nam, Jeong-Hun;Noh, Kyung-Lok;Pang, Eun-O;Yu, Woo-Geun;Kang, Eung-Sun;Kweon, Hae-Yong;Kim, Seong-Gon;Park, Young-Ju
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.366-374
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    • 2010
  • Introduction: This study evaluated the capability of silk fibroin (SF) and recombinant human bone morphogenetic protein-2 loaded SF (SF-BMP) as a bone defect replacement matrix when grafted in a calvarial bone defect of rats in vivo. Materials and Methods: A total 70 calvarial critical size defects (5.0 mm in diameter) made on 35 adult female Sprague-Dawley rats were used in this study. The defects were transplanted with (1) rhBMP-2 loaded silk fibroin graft (SF-BMP: 0.8+$10\;{\mu}g$), (2) Silk fibroin (SF: $10\;{\mu}g$), and (3) no graft material (Raw). The samples were evaluated with soft x-rays, alkaline phosphatase activity, calcium/phosphate quantification, histological and histomorphometric analysis at postoperative 4 and 8 weeks. Results: The SF-BMP group ($48.86{\pm}14.92%$) had a significantly higher mean percentage bone area than the SF group ($24.96{\pm}11.01%$) at postoperative 4 weeks.(P<0.05) In addition, the SF-BMP group ($40.01{\pm}12.43%$) had a higher % bone area at postoperative 8 weeks than the SF group ($33.26{\pm}5.15%$). The mean ratio of gray scale levels to the host bone showed that the SF-BMP group ($0.67{\pm}0.08$) had a higher mean ratio level than the SF group ($0.61{\pm}0.09$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.168 and P=0.243, respectively) The ratio of the calcium and phosphate contents of the SF-BMP ($0.93{\pm}0.22$) group was lower than that of the SF ($1.90{\pm}1.42$) group at postoperative 4 weeks. However, the SF-BMP group ($0.75{\pm}0.31$) had a higher Ca/$PO_4$ ratio than the SF ($0.68{\pm}0.04$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.126 and P=0.627, respectively) For the bone-specific alkaline phosphatase (ALP) activity, which is recognized as a reliable indicator of the osteoblast function, the SF-BMP ($23.71{\pm}8.60\;U/L$) groups had a significantly higher value than the SF group ($12.65{\pm}6.47\;U/L$) at postoperative 4 weeks.(P<0.05) At postoperative 8 weeks, the SF-BMP ($21.65{\pm}10.02\;U/L$) group had a lower bone-specific ALP activity than the SF group ($16.72{\pm}7.35\;U/L$). This difference was not statistically significant.(P=0.263) For the histological evaluation, the SF-BMP group revealed less inflammation, lower foreign body reactions and higher bone healing than the SF group at postoperative 4 and 8 weeks. The SF group revealed more foreign body reactions at postoperative 4 weeks. However, this immunogenic reaction decreased and the remnant of grafted material was observed at postoperative 8 weeks. For histomorphometric analysis, the SF-BMP group had a significantly longer bone length to total length ratio than those of the SF group at postoperative 4 and 8 weeks.(P<0.05) Conclusion: The rhBMP-2 loaded silk fibroin graft revealed fewer immunoreactions and inflammation as well as more new bone formation than the pure silk fibroin graft. Therefore, silk fibroin may be a candidate scaffold for tissue engineered bone regeneration.

The Expression of MUC1 and CD44s in Non-small Cell Lung Cancer (비소세포폐암에서 MUC1과 CD44s의 발현)

  • Park, Hye-Kyung;Lee, Ji-Seok;Lee, Jun-Hee;Lee, Jung-Wook;Kim, Yun-Seong;Lee, Min-Ki;Kim, Young-Dae;Lee, Hyung-Ryu;Kim, Kun-Il;Lee, Chang-Hun;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.2
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    • pp.117-127
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    • 2002
  • Backgroud : MUC1 mucin is a heavily glycosylated large glycoprotein and is expressed aberrantly in carcinoma. CD44 is polymorphic family of cell surface glycoproteins participating in cell-cell adhesion and modulation of the cell-matrix interaction. MUC1 mucin and CD44 expression have been implicated in a tumor invasion and metastasis in certain malignancies. In this study, the expression of MUC1 and the standard form of CD44 (CD44s) was examined in non-small cell lung cancer (NSCLC). Methods : Immunohistochemical staining using monoclonal antibodies including MUC1 glycoprotein and CD44s was performed on 80 NSCLC surgical specimens. The association between MUC1 and CD44s expression and the histological type and tumor stage was investigated. Results : Depolarized MUC1 expression in more than 10% of cancer cells was found in 12 (27.9%) out of 43 squamous cell carcinomas (SCCs) and 12 (32.4%) out of 37 adenocarcinomas (ACs). It was not associated with the tumor histological type and the TNM-stage in SCCs. Depolarized MUC1 expression correlated with the N-stage in ACs (p=0.036). CD44s was expressed in 36 (83.7%) out of 43 SCCs and 14 (37.8%) out of 37 ACs. Reduced CD44s expression correlated with the N-stage (p=0.031) and the TNM-stage (p=0.006) in SCCs. Conclusions : Depolarized MUC1 expression was related to the nodal stage in NSCLC adenocarcinoma. Reduced CD44s expression was related to nodal involvement and the TNM-stage in squamous cell carcinoma. This suggests that MUC1 and CD44s expression in NSCLC might play important roles in tumor progression and cap be used as prognostic variables.

Enzymatically Modified Isoquercitrin Attenuates High-Fat Diet-Induced Obesity (효소 처리된 Isoquercitrin이 고지방식이에 의해 비만이 유도된 마우스의 체중감소에 미치는 영향)

  • Min, Yeojin;Park, Taesun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.474-483
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    • 2016
  • Enzymatically modified isoquercitrin (EMIQ) is a mixture of quercetin glycodsides consisting of isoquercitrin and its ${\alpha}-glucosylated$ derivatives containing one to seven additional linear glucose moieties. The aim of this study was to assess whether or not EMIQ attenuates high-fat diet (HFD)-induced body weight gain and changes in plasma indices of obesity in mice. Male C57BL/6N mice were fed chow diet, HFD, and HFD containing 1.2% EMIQ for 10 weeks. EMIQ significantly (P<0.05) reduced body weight gain (-21%), total visceral fat-pad weights (-31%), and plasma levels of triglycerides (-17%), total cholesterol (-19%), and free fatty acids (-26%) in HFD-fed mice. EMIQ significantly increased protein kinase A (PKA) expression in the epididymal adipose tissue of HFD-fed mice. Expression of adipogenesis-related genes significantly decreased, whereas expression of fatty acid oxidation-related and thermogenesis-related genes increased in epididymal adipose tissue of EMIQ-fed mice compared with HFD-fed mice. These results suggest that the protective effects of EMIQ against HFD-induced adiposity in mice appear to be associated with PKA-mediated signaling cascades involved in adipogenesis, fatty acid oxidation, and thermogenesis in adipose tissue.