• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.22 no.6
• /
• pp.542-549
• /
• 2021

The purpose of this study was to provide an economical and easy preparation method for recombinant human epidermal growth factor (rhEGF) without the need for an expensive enzyme to cleave the fusion part. However, the N-terminal fusion part is still useful for affinity chromatography. The hEGF is an important hormone in cell growth and proliferation in humans, and many studies on the expression and purification of this protein have been reported. In the present study, the hEGF gene was designed to be optimized with the E. coli codon usage preference and to contain Asn-Gly at the N-terminus of the protein. The gene was inserted into pRSET_A, an E. coli expression vector, and transformed into E. coli BL21 (DE3). The recombinant fusion protein was successfully co-expressed with pG-Tf2, a chaperone vector, in E. coli and purified by Ni-NTA column chromatography. The rhEGF was then released by hydroxylamine treatment and confirmed by SDS-PAGE. ELISA analysis showed that the activity of the free rhEGF was more than 92% similar to that of commercial EGF. The biological activity of the rhEGF was confirmed by a cell proliferation test with human skin fibroblasts.

• Microbiology and Biotechnology Letters
• /
• v.42 no.1
• /
• pp.73-81
• /
• 2014

Salsola collina, also known as Russian thistle, is widely distributed in and around waste facilities, roadsides, and drought and semi-drought areas, and is used as a traditional folk remedy in Chinese medicine for the treatment of hypertension. In this study, we have evaluated the anti-oxidative and anti-cancer activities of the ethanol extract of S. collina Pall. (EESC), and the molecular mechanisms of its anti-cancer effects on human colon carcinoma HT29 cells. EESC exhibited anti-oxidative activity through DPPH radical scavenging capacity and showed cytotoxic activity in a dose-dependent manner in HT29 cells. After EESC treatment, HT29 cells altered their morphology, becoming smaller and irregular in shape. EESC also induced cell accumulation in the G2/M phase in a dose-dependent manner, accompanied by a decrease of cell population in the G1 phase. The G2/M arrest by EESC was associated with the increased expression of cyclin-dependent kinase (CDK) inhibitor p21 and Wee1 kinase, which phosphorylates, or inactivates, Cdc2. EESC treatment induced the phosphorylation of Cdc2 and Cdc25C, and inhibited cyclin A and Cdc25C protein expression. In addition, S arrest was induced by the highest concentration of EESC treatment, associated with a decrease of cyclin A and Cdk2 expression. These findings suggest that EESC may possess remarkable anti-oxidative activity and exert an anti-cancer effect in HT29 cells by cell cycle regulation.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.31 no.5
• /
• pp.924-930
• /
• 2002

Ixeris dentata extracts exllibited antimicrobial activity against some bacteria and fungi. Also EtOH extracts showed strong antioxidant activity and RC$_{50}$ value was 28 $\mu\textrm{g}$/mL. The inhibitory effect of Ixeris dentata on the mutagenicity in Salmonella and cytotoxicity on cancer cell were studied. Ixeris dentata extracts showed anti-mutagenic effects of 78.83 and 75.96% on B(a)P in S. typhimurium TA98 and Th100, respectively. These extracts showed 78.72% antimutagenicity on TA100 against MNNG. The Ixeris dentata extract with strong antimutagenic activities was further fractionated by hexane, ethyl acetate, butanol and water. Butanol fraction was found to be highest in antimutagenic activity against MNNG than the other fractions. Butanol fraction of Ixreis dentate revealed the highest cytotoxicity against AS49 human lung carcinoma cells in which cell growth was inhibited by 93.75% at 375 $\mu\textrm{g}$/mL. Hexane fraction of ixeris dentate exhibited 68.56% inhibition against MCF-7 human breast adenocarcinoma cells at 500 $\mu\textrm{g}$/mL. Hexane fraction of Ixeris dentata exhibited 84.91% inhibition against Hep 3B human hepatocellular carcinoma cells at 500 $\mu\textrm{g}$/mL. From these results, it is considered that Ixeris dentata has strong antimutagenic and anticancer effects in vitro. However, these extracts and fractions did not show any cytotoxic effect against 293 cells.

• Asian-Australasian Journal of Animal Sciences
• /
• v.33 no.10
• /
• pp.1624-1632
• /
• 2020

Objective: A total of three hundred unsexed ducks were utilized to estimate net energy requirements of maintenance (NE_m) and weight gain (NE_g) for 2 to 3-week-old Cherry Valley ducks and to establish a model equation to predict NE requirements using the factorial method. Methods: To determine the apparent metabolizable energy (AME) of the diet, fifty 7-day-old ducks at approximately equal body weights (BWs) were randomly assigned into five groups that were fed at different levels (ad libitum, 85%, 75%, 65%, and 55% of ad libitum intake), and the endogenous acid-insoluble ash as indigestible marker. The two hundred and fifty 7-day-old ducks were used for a comparative slaughter experiment. At the beginning of the experiment, ten ducks were sacrificed to determine the initial body composition and energy content. The remaining ducks were randomly assigned into five groups (same as metabolic experiment). Ducks of the ad libitum group were slaughtered at 14 and 21-day-old. At the end of the experiment, two ducks were selected from each replicate and slaughtered to determine the body composition and energy content. Results: The results of the metabolizable experiment showed AME values of 13.43 to 13.77 MJ/kg for ducks at different feed intakes. The results of the comparative slaughter experiment showed the NE_m value for 2 to 3-week-old Cherry Valley ducks was 549.54 kJ/kg of BW^0.75/d, and the NE_g value was 10.41 kJ/g. The deposition efficiency values of fat (K_f) and crude protein (K_p) were 0.96 and 0.60, respectively, and the values of efficiency of energy utilization (K_g) and maintenance efficiency (K_m) were 0.75 and 0.88, respectively. Conclusion: The equation for the prediction of NE requirements for 2 to 3-week-old Cherry Valley ducks was the following: NE = 549.54 BW^0.75+10.41 ΔW, where ΔW is the weight gain (g).

• Korean journal of food and cookery science
• /
• v.28 no.6
• /
• pp.711-719
• /
• 2012

The collagens from squid (Todarodes pacificus) skin and Alaska pollack (Theragra chalcogramma) skin were extracted and their physicochemical properties were investigated. The yield was improved with the increase of NaOH concentration and was increased Neutrase than Alcalase in enzymatic hydrolysates. Protein and collagen contents from Alaska pollack skin were 38.3~62.7% and 13.1~28.9%, respectively. All enzymatic hydrolysates also showed high antioxidant activities as NaOH concentration decrease. Composition of their amino acids was mainly glycine and proline. The spectrum of FT-IR of the collagen showed wavenumber at $1,631cm^{-1}$, $1,549cm^{-1}$, $1,234cm^{-1}$ and $3,322cm^{-1}$ representing the regions of amide I, amide II, amide III and amide A, respectively. The decomposition temperature for the collagen was in the range of $300^{\circ}C$ and showed relatively good enough for their thermal stabilities.

• Analytical Science and Technology
• /
• v.15 no.6
• /
• pp.540-549
• /
• 2002

The arrangement of encapsulated Ar atoms in the molecular-dimensioned cavities of fully dehydrated zeolite A of unit-cell composition $Cs_3Na_8HSi_{12}Al_{12}O_{48}$ ($Cs_3$-A) has been studied crystallographically to probe the confinement effect of guest species in microporous environment. Atoms of Ar were encapsulated in the cavities of $Cs_3$-A by treatment with 410 atm of Ar at $400^{\circ}C$ for two days, followed by cooling at room temperature. The crystal structure of $Cs_3Na_8H$-A(4Ar) ($P_e$ = 410 atm, $a=12.245(2){\AA}$, $R_1=0.0543$, and $R_2=0.0552$) has been determined by single crystal X-ray diffraction technique in the cubic space group $Pm\bar{3}m$ at 21 (1) $^{\circ}C$ and 1 atm. Encapsulated Ar atoms are distributed in three crystallographic distinct positions: 1.5 Ar atoms per unit cell opposite 6-rings, 1.5 opposite four-rings in the large cavity, and finally 1.0 in the sodalite-unit. The possible structures of argon clusters, such as $Ar_2$, $Ar_3$, and $Ar_4$, are proposed.

• Journal of Life Science
• /
• v.21 no.4
• /
• pp.549-553
• /
• 2011

This study was performed to determine whether or not urinary 1-hydroxypyrene (1-OHP) levels can be accurately detected by our 1-OHP-detecting $TiO_2$-Bead-HPLC assay that we developed based on the molecular imprinting method. Our method showed a variation coefficient of 4.97% and a between-day variation coefficient of 4.43%, suggesting that this may be a very stable method. In addition, the recovery rate of 1-OHP from a mixture of 1-OHP and similar substances using our $TiO_2$-Bead-HPLC method was estimated to be 105.6%. The correlation coefficient between the conventional enzyme-HPLC method and this new method was 0.74 (p<0.01) when the urine samples were tested. Based on this result, it is conceivable that our method could be a useful technique for measuring urinary 1-OHP levels. Moreover, our method has some advantages of being easier and less expensive than the conventional method. The results of this study suggest that our method can facilitate the development of a urine 1-OHP sensor using $TiO_2$-coating beads and that development of beads by molecular imprinting can be applied to analysis of chemicals other than 1-OHP.

• Tuberculosis and Respiratory Diseases
• /
• v.57 no.5
• /
• pp.449-460
• /
• 2004

Background : PS-341 is a novel, highly selective and potent proteasome inhibitor, which showed cytotoxicity against some tumor cells. Its anti-tumor activity has been suggested to be associated with modulation of the expression of apoptosis-associated proteins, such as p53, $p21^{WAF/CIP1}$, $p27^{KIP1}$, NF-${\kappa}B$, Bax and Bcl-2. c-Jun N-terminal kinase (JNK) and glycogen synthase kinase-$3{\beta}$ (GSK-$3{\beta}$) are important modulators of apoptosis. However, their role in PS-341-induced apoptosis is unclear. This study was undertaken to elucidate the role of JNK and GSK-$3{\beta}$ in the PS-341-induced apoptosis in lung cancer cells. Method : NCI-H157 and A549 cells were used in the experiments. The cell viability was assayed using the MTT assay and apoptosis was evaluated by proteolysis of PARP. The JNK activity was measured by an in vitro immuno complex kinase assay and by phosphorylation of endogenous c-Jun. The protein expression was evaluated by Western blot analysis. Dominant negative JNK1 (DN-JNK1) and GSK-$3{\beta}$ were overexpressed using plasmid and adenovirus vectors, respectively. Result : PS-341 reduced the cell viability via apoptosis, activated JNK and increased the c-Jun expression. Blocking of the JNK activation by overexpression of DN-JNK1, or pretreatment with SP600125, suppressed the apoptosis induced by PS-341. The activation of caspase 3 was mediated by JNK activation. Blocking of the caspase 3 activation suppressed PS-341-induced apoptosis. PS-341 activated the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, but its blockade enhanced the PS-341-induced cell death via apoptosis. GSK-$3{\beta}$ was inactivated by PS-341 via the PI3K/Akt pathway. Overexpression of constitutively active GSK-$3{\beta}$ enhanced PS-341-induced apoptosis; in contrast, this was suppressed by dominant negative GSK-$3{\beta}$ (DN-GSK-$3{\beta}$). Inactivation of GSK-$3{\beta}$ by pretreatment with lithium chloride or the overexpression of DN-GSK-$3{\beta}$ suppressed both the JNK activation and c-Jun up-regulation induced by PS-341. Conclusion : The JNK/caspase pathway is involved in PS-341-induced apoptosis, which is negatively regulated by the PI3K/Akt-mediated inactivation of GSK-$3{\beta}$ in lung cancer cells.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.10
• /
• pp.4331-4338
• /
• 2014

$N^1$-(2, 5-dimethoxyphenyl)-$N^8$-hydroxyoctanediamide (N25) is a novel SAHA cap derivative of HDACi, with a patent (No. CN 103159646). This invention is a hydroxamic acid compound with a structural formula of $RNHCO(CH_2)6CONHOH$ (wherein R=2, 5dimethoxyaniline), a pharmaceutically acceptable salt which is soluble. In the present study, we investigated the effects of N25 with regard to drug distribution and molecular docking, and anti-proliferation, apoptosis, cell cycling, and $LD_{50}$. First, we designed a molecular approach for modeling selected SAHA derivatives based on available structural information regarding human HDAC8 in complex with SAHA (PDB code 1T69). N25 was found to be stabilized by direct interaction with the HDAC8. Anti-proliferative activity was observed in human glioma U251, U87, T98G cells and human lung cancer H460, A549, H1299 cells at moderate concentrations ($0.5-30{\mu}M$). Compared with SAHA, N25 displayed an increased antitumor activity in U251 and H460 cells. We further analyzed cell death mechanisms activated by N25 in U251 and H460 cells. N25 significantly increased acetylation of Histone 3 and inhibited HDAC4. On RT-PCR analysis, N25 increased the mRNA levels of p21, however, decreased the levels of p53. These resulted in promotion of apoptosis, inducing G0/G1 arrest in U251 cells and G2/M arrest in H460 cells in a time-dependent and dose-dependent manner. In addition, N25 was able to distribute to brain tissue through the blood-brain barrier of mice ($LD_{50}$: 240.840mg/kg). In conclusion, our findings demonstrate that N25 will provide an invaluable tool to investigate the molecular mechanism with potential chemotherapeutic value in several malignancies, especially human glioma.

• Korean Journal of Head & Neck Oncology
• /
• v.16 no.2
• /
• pp.177-181
• /
• 2000

배경 및 목적: Theodor Kocker에 의해 일반화된 전통적인 갑상선 수술방법은 갑상선 질환의 종류 및 정도, 수술범위와 상관없이 광범위한 수술범위로 인한 조직 손상으로 인해 수술후 환자들의 여러가지 불편감은 물론 경부의 넓은 부위의 통증과 경부 피부부종, 장액종, 혈종 등과 같은 후유증을 동반할 수 있다. 최근 본 저자들은 이같은 전통적 갑상선 수술의 부작용을 최소화하기 위해 작은 피부절개($3{\sim}4.5cm$) 후 피하 피판(subplatysmal skin flap) 없이 직접 갑상선으로 접근하는 새로운 수술기법으로서 최소침습 갑상선 수술기법을 개발하였기에 그 술식을 소개하고 전통적인 갑상선 절제술에 대한 우월성을 확인하고자 본 연구를 시행하였다. 대상 및 방법: 1999년 1월 15일 부터 2000년 1월 14일까지 573예의 갑상선 수술 예 중 최소침습 갑상선절제술이 시행되었던 466예와 1998년 1월 15일부터 1999년 1월 14일까지 전통적 갑상선 수술을 시행한 549예 중 거대 종양(양성>6cm, 악성>5cm), 흉골하 선종, 국소진행암, 재발암, 측경부의 다발성 림프절 전이가 있었던 112예를 제외한 437예의 임상병리적 특성과 피부절개 길이, 수술 시간, 수술중 출혈양, 수술후 진통제 요구빈도 및 재윈기간, 수술 후 합병증 발생빈도를 비교 분석하였다. 결 과: 두 군간의 임상병리적 특성상의 유의한 차이는 없었다. 피부절개 길이($3.7{\pm}0.7cm,\;vs\;9.6{\pm}3.3cm$), 수술 시간($57.6{\pm}11.7$분 vs $85.2{\pm}32.3$분) 수술 중 출혈양($18.4{\pm}15.3ml\;vs\;43.1{\pm}21.8ml$), 수술후 재원기간($1.6{\pm}0.5$일 vs $4.3{\pm}1.6$일), 및 수술후 진통제 요구빈도가 전통적 수술군에 비해 최소침습 수술군에서 통계적으로 유의하게 감소되었으나(p<0.05), 수술후 장액종 및 혈종 형성, 일시적인 음색변화, 일시적인 저칼슘혈증과 같은 합병증의 발생빈도는 각각 4.3%(n=20)와 4.8%(n=21)로 두 군간에 유의한 차이가 없었다. 결 론: 최소침습 갑상선 수술법은 새로운 수술기구의 도입 없이도 갑상선 수술의 충분한 시야를 확보할 수 있고 안전하고 간단하게 시행할 수 있으며, 기존 수술법으로 인한 부작용을 최소화할 수 있어 전통적 인 수술법을 대치할 수 있는 새로운 방법으로 사료된다.