• 한국식품영양학회지
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• 제25권4호
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• pp.800-806
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• 2012

체내에서 산화스트레스에 의해 생성되는 활성산소종(reacitve oxygen species, ROS)은 당뇨병, 비만 등과 같은 만성질환을 야기시키는 것으로 알려져 있다. 고려인삼(Panax ginseng)은 수천 년간 피로 회복 및 면역증강용 기능성식품으로 이용되어 왔고, 사포닌, 산성다당체, 페놀성 화합물과 같은 다양한 생리활성 물질을 함유하고 있다. 따라서 본 연구에는 고온, 고압처리하여 제조한 신규 인삼에 대한 산화적 스트레스 저감 효능을 평가하고자 하였다. C2C12 근육세포에 산화적 스트레스를 유도하기 위해 $H_2O_2$ 1 mM 처리하고, 전처리 조건을 달리한 인삼 시료를 처리하여 cell morphology 및 항산화 관련 유전자인 SOD, CAT 및 GPx를 살펴보았고, 3T3-L1 지방세포는 분화과정 중 ROS 생성 억제효과 및 CAT, GPx 및 Cu/Zn-SOD의 항산화효소 관련 유전자의 발현 정도를 조사하였다. 고온, 고압처리한 인삼은 산화적 스트레스가 유도된 C2C12 근육세포 및 3T3-L1 지방세포에서 유의적으로 산화적 스트레스를 저감하는 것으로 나타났다. 이상의 결과로 보아, 본 연구진에 의해 개발된 고온 및 고압 처리된 인삼은 항산화 및 항피로 효능이 기대되는 바이며, 본 연구는 동물세포 수준에서의 비교이며, 보다 정확한 작용기전의 구명을 위해 향후 추가적인 연구를 통한 비교 실험이 수행되어야 할 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5421-5425
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• 2013

Background: To assess the antioxidant effects of gamma-oryzanol on human prostate cancer cells. Materials and Methods: Cytotoxic activity of gamma-oryzanol on human DU145 and PC3 prostate cancer cells was determined by proliferation assay using 3-(4, 5-dimethylthiazol, 2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) reagent. mRNA levels of genes involved in the intracellular antioxidant system, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GSR) were determined by reverse transcription-polymerase chain reaction (RT-PCR). Cancer cell lysates were used to measure lipid peroxidation using thiobarbituric acid reactive substance (TBARS). Glutathione contents of the cell lysates were estimated by the reaction between sulfhydryl group of 5, 5'-dithio (bis) nitrobenzoic acid (DTNB) to produce a yellow-color of 5-thio-2-nitrobenzoic acid using colorimetric assay. Catalase activity was also analysed by examining peroxidative function. Protein concentration was estimated by Bradford's assay. Results: All concentrations of gamma-oryzanol, 0.1-2.0mg/ml, significantly inhibited cell growth in a dose- and time-dependent fashion in both prostate cancer cell lines, DU145 and PC3. Gene expression of catalase in DU145 and PC3 exposed to gamma-orizanol at 0.5mg/ml for 14 days was down regulated, while mRNA of GPX was also down regulated in PC3. The MDA and glutathione levels including catalase activity in the cell lysates of DU145 and PC3 treated with gamma-oryzanol 0.1 and 0.5mg/ml were generally decreased. Conclusions: This study highlighted effects of gamma-oryzanol via the down-regulation of antioxidant genes, catalase and GPX, not cytotoxic roles. This might be interesting for adjuvant chemotherapy to make prostate cancer cells more sensitive to free radicals. It might be useful for the reduction of cytotoxic agents and cancer chemoprevention.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.347-355
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• 2016

An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 was heterologously expressed and purified. The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. AorCsxA exhibited optimal activity at pH 5.5 and 50℃; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50℃ than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. AorCsxA activity was inhibited by EDTA and most of the tested metal ions. A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of V_max and k_cat values with those of the wild-type enzyme. The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA.

• Journal of Genetic Medicine
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• 제12권1호
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• pp.44-48
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• 2015

Townes-Brocks syndrome (TBS) is a rare genetic disorder characterized by the classic triad of congenital anomalies of the anus, thumbs, and ears, with variable expressivity. Additionally, renal malformations, cardiac anomalies, and endocrine and eye abnormalities can accompany TBS, although less frequently. TBS is inherited in an autosomal dominant fashion; however, about 50% of patients have a family history of TBS and the remaining 50% have de novo mutations. SALL1, located on chromosome 16q12.1, is the only causative gene of TBS. SALL1 acts as a transcription factor and may play an important role in inducing the anomalies during embryogenesis. Clinical features of TBS overlap with those of other multiple anomaly syndromes, such as VACTERL syndrome, Baller-Gerold syndrome, Goldenhar syndrome, cat eye syndrome, and Holt-Oram syndrome. Consequently, there are some difficulties in differential diagnosis based on clinical manifestations. Herein, we report a Korean family with two generations of TBS that was diagnosed based on physical examination findings and medical history. Although the same mutation in SALL1 was identified in both the mother and the son, they displayed different clinical manifestations, suggesting a phenotypic diversity of TBS.

• 한국식품영양과학회지
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• 제41권4호
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• pp.437-442
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• 2012

홍국쌀 추출물의 항산화 작용을 DPPH radical 및 hydroxyl radical 소거능, 간암 세포에서의 산화적 DNA 손상과 항산화 효소의 유전자 발현에 미치는 영향으로 분석하였다. 홍국쌀 추출물의 DPPH radical 소거능은 ethyl acetate 추출물, methanol 추출물, butanol 추출물 순이었으며, ethyl acetate 추출물의 DPPH radical 소거능은 0.2 mg/mL에서 85%, $IC_{50}$는 0.13 mg/mL로 나타났다. Ethyl acetate 추출물의 hydroxyl radical 소거능을 DCF fluorescence로 측정한 결과, cuvette에서는 2.5 ${\mu}g$/mL에서 44.2%, 5.0 ${\mu}g$/mL 74.1%, 10.0 ${\mu}g$/mL >100%로 나타났고, $HepG_2$ cell에서는 ethyl acetate 추출물로 전처리한 세포의 radical이 $H_2O_2$로만 처리한 세포에 비해 유의적으로 감소하였다. 홍국쌀 ethyl acetate로 전처리한 세포의 DNA 손상이 $H_2O_2$로만 처리한 세포에 비해 유의적으로 낮았으며, 추출물 대신 lovastatin을 처리한 세포는 DNA 손상이 증가하는 것으로 나타났다. 항산화 효소 유전자의 상대적 발현 정도를 측정한 결과, 산화 스트레스 없이 ethyl acetate 추출물로 전처리한 세포에서는 SOD와 GPx가 control에 비해 각각 3.25배, 2.67배 유의적으로 증가하였으며, 추출물로 전처리한 후 산화 스트레스에 노출시킨 세포에서는 CAT가 control에 비해 5 ${\mu}g$/mL에서 4.64배, 10 ${\mu}g$/mL에서 7.0배 유의적으로 증가하였다.

• 한방재활의학과학회지
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• 제21권4호
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• pp.23-40
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• 2011

Objectives: This study was designed to examine the effects of extracts of Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) on the lipid lowering, anti-oxidation and concentration of proinflammatory cytokines and was investigated on hyperlipidemic rats. Methods: Male rats weighing $182.39{\pm}4.71g$ were fed high fat diet for 8 weeks and 36 rats(above 400 g) were divided into 4 groups. Each of 9 rats was divided a control group and experimental groups. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And we fed each experimental group of rats basal diet and administered an extract of Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) extracts(100 mg/kg, 200mg/kg, 300 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid of plasma and liver, concentration of proinflmmatory cytokines, anti-oxidative activity and $TNF-{\alpha}$, Apo-B, Apo-E and leptin gene expression. Results: 1. Concentration of plasma free fatty(FFA) showed no significant difference in all the treatment groups. Concentration of plasma triglyceride(TG) showed a significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. 2. Concentration of plasma total cholesterol showed a significant decrement in the 200 and 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. Concentration of plasma low density lipoprotein(LDL)-cholesterol showed a Significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. Concentration of plasma high density lipoprotein(HDL)-cholesterol showed a significant increment in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group. 3. Concentration of liver total cholesterol showed a tendence to decrease in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups. Concentration of liver TG showed a significant decrement in all Ojeoksangamibang groups than that of control group. 4. Concentration of plasma and liver thiobarbituric acid reactive substance(TBARS) showed a tendence to decrease in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups. 5. The values of glutathione peroxidase(GSH-Px), superoxide dismutase(SOD) and catalase(CAT) activity showed a significant increment in all Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. 6. The values of plasma aspartate aminotransferase(AST) and alanine aminotransferase(ALT) activity showed no significant different in all treatment group. 7. Concentration of plasma $interleukin(IL)-1{beta}$ showed no significant difference in all the treatment groups. Concentration of plasma IL-6 showed a significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group than that of control group. Concentration of plasma tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ a siginifant decrement in the 200 and 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group than that of control group. However the concentration of plasma IL-10 in the 300 mg/kg Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a significant increment than that of control group. 9. In the analysis of reverse transcription-polymerase chain reaction(RT-PCR), gene expression of $TNF-{\alpha}$, Apo-B and Apo-E in the Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a lower expression than that of control group. However the gene expression of leptin showed no difference in the treatment groups. 10. The ratio of $TNF-{\alpha}$, Apo-B, and Apo-E per ${\beta}-actin$ expression in the Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a significant decrement than that of control group. However The ratio of leptin expression per ${\beta}-actin$ expression showed no significant difference among all the treatment groups. Conclusions: According to above results, in lowering lipid effect, anti-oxidation and control of pro-inflammatory cytokines production, Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) gives effect.

• 대한본초학회지
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• 제33권4호
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• pp.9-18
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• 2018

Objectives : This study aimed to investigate the anti-ulcer effect of an standardized herbal extracts mixture of Inulae Flos and Paeoniae Radix (HT074) on acidified ethanol induced gastric injury and its potential mechanisms. Methods : Antioxidant activities of HT074 and its constituents were measured by DPPH (2,2-Diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging capacity. After the oral administration of HT074 at doses of 100, 300 mg/kg twice per day for 14 days, Gastric lesions were induced by oral administration of acidified ethanol in Sprague Dawley rats. Oxidative stress markers, such as super oxide dismutase (SOD) activity, concentrations of catalase (CAT) and glutathione (GSH) were measured in gastric mucosal tissues. Additionally, the expression of human mucin gene, Mucin 5AC (MUC5AC) mRNA in gastric mucosal tissues was measured. Results : HT074 showed dose dependent radical scavenging activities against DPPH and ABTS radicals. Oral administration of HT074 300 mg/kg for 14 consecutive days significantly decreased gastric lesions and histological damages induced by HCl/EtOH in rats. HT074 treatment significantly increased the activity of SOD (300 mg/kg) and concentration of GSH (100 and 300 mg/kg), however catalase concentration was not significantly increased. MUC5AC mRNA expression was significantly increased by HT074 100, 300 mg/kg treatment. Conclusions : HT074 protects the gastric mucosa from oxidative stress caused by acidified ethanol by increasing the activity of SOD, concentration of GSH and mucin biosynthesis. These findings suggest that HT074 could be an effective candidate for prevention and treatment of gastritis and gastric ulcer.

• Tuberculosis and Respiratory Diseases
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• 제49권4호
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• pp.453-465
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• 2000

연구배경 : 폐암의 발생과정은 다양한 유전자 이상과 여러 가지경로 이상을 포함한 다단계 과정이다. 암유전자의 활성화나 종양억제유전자의 불활성화, 그리고 결과적인 유전적 불안정성의 증가는 폐암의 발암과정에서 일어나는 주요한 사건이며 임상적으로 폐암이 진단되기까지 10내지 20여 가지의 유전적 변화가 축적되는 것으로 알려져 있다. 본 연구에서 저자들은 비소세포폐암에서 종양억제유전자인 p53과 FHIT의 돌연변이, FHIT 유전자의 전사체 이상 여부를 확인하고 종양억제유전자부근에 위치하는 극소위성의 유전적 변화를 관찰하였다. 대상 및 방법 : 비소세포폐암으로 진단된 후 외과적 적출술을 시행받은 환자 29명의 생검조직과 그에 대응하는 동일인의 정상조직을 대상으로 하였다. p53과 FHIT의 돌연변이 여부는 PCR-SSCP, DNA 염기분석으로 확인하였고 D3S1285, D9S171, TP53에서 극소위성 불안정성과 이형접합성 상실은 PCR로 확인하였다. FHIT 유전자의 전사체 이상 여부 확인을 위해서는 RT-PCR을 사용하였다. 결과 : 1) p53 유전자의 2예에서 관찰되었고 모두 exon 5에서 1개의 염기가 치환되는 점돌연변이였다. 2) 극소위성 불안정성은 D3S1285와 D9S171에서 각각 2예, 1예, 이형접합성 상실은 D3S1285, D9S171, TP53에서 각각 3예, 4예, 7예가 관찰되었다. 3) FHIT 유전자의 변이는 11예에서 관찰되었으며 이중 6예는 exon 8의 codon 98에서 염기서열이 CAT가 CAC로 바뀌는 잠재적 치환이었다. 4) FHIT 유전자의 전사체 이상은 $\beta$-actin이 제대로 발현되는 15예중 4예에서 관찰되었으며 exon 6-9의 결실로 확인되었다. 결론 : 이상으로 비소세포폐암 발생에 p53, FHIT 유전자의 변이, 극소위성 불안정성과 이형접합성 상실 등 다양한 분자유전학적 기전이 복합적으로 작용할 것으로 생각되며 이번 연구에서 조사된 유전적 이상의 빈도는 앞서 발표된 서양의 연구결과와 대체적으로 일치한다. 특히 극소위성의 분석은 편평세포암에서 종양표지자로서의 역할이 기대된다. 이런 발암과정에 대한 이해는 예방, 진단 및 치료적 접근을 발전시키는데 도움을 줄 수 있을 것이고 향후 이들에 관한 가능적 연구들이 수행되어야 할 것이다.

• 한국식품저장유통학회지
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• 제21권2호
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• pp.264-275
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• 2014

본 연구에서는 화장품 조성물 내에 함유된 야관문 추출물의 자외선 조사에 의한 피부 광노화를 억제 및 개선할 수 있는 효능을 조사하였다. 야관문 추출물 내 총 폴리페놀 함량은 $134.98{\pm}1.70$ mg/g, 총 플라보노이드 함량은 $16.20{\pm}0.05$ mg/g 으로 정량되었다. 야관문 추출물은 농도 의존적으로 전자공여능 및 자유라디칼 소거능에 의한 항산화 활성을 나타내는 것을 확인하였으며 동일 농도조건에서 전자공여능 보다 자유라디칼 소거능에 의한 항산화력이 더욱 높았다. 피부 홍반도는 대조군(CO)에 비하여 야관문 추출물이 함유된 시료를 도포한 시험군(AS)에서 28%의 홍반도 감소 효과를 보였으며, 피부 수분함량은 AS군에서 CO군과 비교하여 유의하게 증가하였다. 피부주형의 형태학적 관찰을 통해 AS군에서 피부의 주름 면적과 깊이가 현저하게 감소하였음을 확인하였으며, 조직염색을 통해 AS군에서 자외선 조사에 의해 손상된 피부조직이 정상적인 피부구성 조직과 유사하게 재생되었음을 확인하였다. 피부조직 내 유해산소 해독계 효소들인 SOD, GST와 CAT 활성은 CO군과 비교해 AS군에서 유의적인 증가를 보였으며, 유해산소 생성계 효소 XO와 지질과산화물 함량은 유의하게 감소하였다. 자외선 노출에 의해 발현이 유도되는 PAK, p38, c-Fos, c-Jun, TNF-${\alpha}$, MMP-3 유전자들은 CO군 대비 AS군에서 그 발현이 유의하게 감소함을 확인하였다. 이와 같은 결과는 야관문 추출물은 항산화능을 나타내는 생리활성물질의 작용을 통해 피부 광노화에 의한 피부손상과 주름형성을 개선하는데 분자적 수준에서 형태학적 수준까지 효능을 나타내어 피부 광노화억제 및 개선 제품으로 활용이 가능 할 것으로 기대된다.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1085-1091
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• 2009

A phytase with high activity at low temperatures has great potential for feed applications, especially in aquaculture. Therefore, this study used a degenerate PCR and TAIL PCR to clone a phytase gene from Erwinia carotovora var. carotovota, the cause of soft rot of vegetables in the ground or during cold storage. The full-length 2.5-kb fragment included an open reading frame of 1,302 bp and encoded a putative phytase of 45.3 kDa with a 50% amino acid identity to the Klebsiella pneumoniae phytase. The phytase contained the active site RHGXRXP and HD sequence motifs that are typical of histidine acid phosphatases. The enzyme was expressed in Escherichia coli, purified, and displayed the following characteristics: a high catalytic activity at low temperatures (retaining over 24% activity at $5^{\circ}C$) and remarkably thermal lability (losing >96% activity after incubation at $60^{\circ}C$ for 2 min). The optimal phytase activity occurred at pH 5.5 and ${\sim}49^{\circ}C$, and the enzyme activity rapidly decreased above $40^{\circ}C$. When compared with mesophilic counterparts, the phytase not only exhibited a high activity at a low temperature, but also had a low $K_m$ and high $k_{cat}$. These temperature characteristics and kinetic parameters are consistent with low-temperature-active enzymes. To our knowledge, this would appear to be the first report of a low-temperature-active phytase and its heterogeneous expression.