• Title/Summary/Keyword: in vitro

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Hypoglycemic Effects of Fruits and Vegetables in Hyperglycemic Rats for Prevention of Type-2 Diabetes (고혈압쥐의 과일과 야채의 섭취에 따른 저혈당 효과)

  • Survay, Nazneen Shaik;Ko, Eun-Young;Upadhyay, Chandrama Prakash;Jang, Mi;Park, Se-Won;Lee, Dong-Ha;Jung, Yi-Sook;Yoon, Do-Young;Hong, Sae-Jin
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.850-856
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    • 2010
  • An in vivo oral glucose tolerance test (OGTT) was performed on hyperglycemic male Sprague-Dawley rats to assess the effect of fruits and vegetables ($1g{\cdot}kg^{-1}$ body weight) on blood glucose levels (${\Delta}BGLs$) at different time intervals of 0, 5, 15, 30, 60, 90 and 120 min. The areas under glucose curve (${\Delta}AUCs$) were calculated at 120 min of OGTT by trapezoid method. Total phenolic content (TPC) and anti-oxidant activity (AOA) of fruits and vegetables were assayed in vitro by Folin Ciocalteu and DPPH (2, 2-diphenyl-1-picrylhydrazyl) methods, respectively. At the end of the experiment the correlations among the parameters TPC, AOA and ${\Delta}AUC$ was estimated by Pearson's correlations. Among fruit crops, tangerine, plum, grape and pear and among vegetables, blue leaf mustard, cabbage, chicory, broccoli and others exhibited significant hypoglycemic effects by reducing ${\Delta}BGLs$ with significant ${\Delta}AUC$. The effective ${\Delta}AUC$ ranged from $5548.2{\pm}462.1$ to $3823.3{\pm}282.0mg-min{\cdot}dL^{-1}$. The TPC and AOA ranged from $0.063{\pm}0.00$ to $0.913{\pm}0.14mg{\cdot}g^{-1}$ GAE and $01.05{\pm}0.08$ to $75.46{\pm}0.06%$, respectively. Overall, six fruits and fifteen vegetables exhibited higher TPC and one fruit and four vegetables exhibited higher AOA. There was a better correlation among TPC, AOA and ${\Delta}AUC$ of fruits and TPC & AOA of vegetables. We report that hypoglycemically significant fruits and vegetables investigated in this study have pharmacological importance which reduced ${\Delta}BGLs$ through insulin like activity and AOA in prevention of type-2 diabetes.

Enhancement of the Thermostability of a Fibrinolytic Enzyme from Bacillus amyloliquefaciens CH51 (Bacillus amyloliquefaciens CH51이 생산하는 혈전용해효소의 열안정성 개선)

  • Kim, Jieun;Choi, Kyoung-Hwa;Kim, Jeong Hwan;Song, Young-Sun;Cha, Jaeho
    • Journal of Life Science
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    • v.23 no.1
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    • pp.15-23
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    • 2013
  • AprE51 from Bacillus amyloliquefaciens CH51 is a 27 kDa subtilisin-like protease with fibrinolytic activity. AprE51-6 showing increased catalytic activity was produced previously. To enhance the thermostability of AprE51-6, 2 residues, Gly-166 and Asn-218 based on B. subtilis subtilisin E were mutated by site-directed mutagenesis. The results of the mutational analysis showed that substitution of arginine for Gly-166 (AprE51-7) increased the fibrinolytic activity 1.8-fold. An N218S mutant (AprE51-8) also increased the fibrinolytic activity up to 4.5-fold in a fibrin plate assay. Purified AprE51-7 and AprE51-8 mutants had a 1.9- and a 2.5-fold higher $k_{cat}$, respectively, and a 2.1-1.9-fold lower $K_m$, respectively. This resulted in a 3.8- and a 4.7-fold increase in catalytic efficiency ($k_{cat}/K_m$), respectively, relative to that of wild-type AprE51. AprE51-8 had a broader pH range than AprE51-6 and nattokinase, especially at an alkaline pH value. In addition, AprE51-8 showed higher thermostability than AprE51-6 at $60^{\circ}C$. The half-lives of AprE51-7 and AprE51-8 at $50^{\circ}C$ were 21.5 and 27.3 min, respectively, which are 2.0 and 2.6 times longer, respectively, than that of the wild-type AprE51.

Removal of Dissolved Organic Matter by Ozone-biological Activated Carbon process (오존처리와 생물활성탄 공정에 의한 상수원수 중의 용존유기물 제거)

  • 이상훈;문순식;신종철;최광근;심상준;박대원;이진원
    • KSBB Journal
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    • v.18 no.3
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    • pp.211-216
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    • 2003
  • The removal yield of dissolved organic matter in drinking water by biological activated carbon (BAC) process was investigated. The tested processes wer raw water-AC process (BAC1), raw water-ozonation-BAC process (BAC2), and raw water-ozonation-coagulation/sedimentation-BAC process (BAC3). The amounts of organic matter was measured as dissolved organic carbon (DOC), ulta-violet radiation at 254 nm wavelength ($UV_{254}$), total nitrogen (T-N), ammonia nitrogen (NH_3$-N), and total phosphate (T-P). As a results, 30.7% DOC was removed by BAC2 process, which showed higher removal efficiency than BAC1 or BAC3 processes. The removal yield of $UV_{254}$ in BAC1, BAC2, and BAC3 processes were observed as 45.3%, 44.6%, 58.4%, respectively. And the removal yield of ammonia nitrogen were 66%, 81%, 29% in each BAC processes. The optimal empty bed contact time (EBCT) of BAC processes was estimated as 10 minute. This study has shown that BAC process combined with ozone treatment was efficient for removing dissolved organic matter in water.

EFFECT OF HYDROGEN PEROXIDE CONCENTRATION ON THE WHITENING AND PHYSICAL PROPERTIES OF HYDROXYAPATITE DISCS (Hydrogen Peroxide 농도와 적용시간이 Hydroxyapatite Discs의 미백과 물리적 성질에 미치는 영향)

  • Yang, Yeon-Mi;Lee, Doo-Cheol;Baik, Byeong-Ju;Kim, Jae-Gon;Shin, Jeong-Geun
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.1
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    • pp.1-12
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    • 2007
  • The purpose of this study was to evaluate the effect that various concentration and application time of hydrogen peroxide had on tooth whitening and physical properties. The hydroxyapatite (HA) discs of $12mm({\Phi}){\times}1.2mm(t)$ in dimensions were made by compression $(100kg/cm^2)$ and sintering (at $1350^{\circ}C$ for 2 hours) All specimens were polished sequentially with '240 through '2000 emery paper and one side of each specimen was polished finally with $0.3{\mu}m$ alumina paste. The discs were placed in sterile whole stimulated saliva overnight at $37^{\circ}C$ in order to form an in vitro pellicle layer. Then the discs were rinsed with distilled water and soaked into staining broth at $37^{\circ}C$ for 7 days. These stained specimens were bleached with hydrogen peroxide according to the change of concentration $(3{\sim}30%)$ and application time ($3{\sim}10$ days). The specimens were analyzed with a spectrophotometer, X-ray diffractometer (XRD), scanning electron microscope (SEM), surface roughness tester, microhardness tester and biaxial flexural strength. The results of present study can be summarized as follows : 1. The bleaching effect was increased with the increased concentration and the extended application time of hydrogen peroxide. 2. The surface roughness was significantly increased from the specimen bleached with 15% hydrogen peroxide for 10 days and with 30% for 7 and 10 days respectively (p<0.05). 3. The changes of crystal phase observed by XRD between before and after bleaching weren't shown of any difference, but microporous structure of surface observed by SEM was shown of increase with the increased concentration and the extended application. 4. The biaxial flexural strength was significantly decreased from bleaching of specimen with 30% hydrogen peroxide for 7 and 10 days respectively (p<0.05) 5. Microhardness was significantly decreased from bleaching with 15% hydrogen peroxide for 10 days and with 30% for 3, 7 and 10 days respectively (p<0.05). Although the tooth bleaching effect was greater when the high concentration was applied, further in vivo experiment will be needed to prove it's safety.

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Effect of Dexamethasone on Gene Expression of Surfactant Protein B and Surfactant Protein C (스테로이드제가 백서 폐의 Surfactant B와 C 유전자 발현에 미치는 영향)

  • Park, Ik Soo;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.4
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    • pp.439-448
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    • 2003
  • Background : Surfactant protein B(SP-B) and surfactant protein C(SP-C) are important in accelerating surface spreading of surfactant phospholipid. The glucocorticoids accelerate the morphologic differentiation of epithelial cells into type II cells and increase the rate of phosphatidylcholine synthesis. The hydrophobic surfactant protein has been shown to be upregulated by glucocorticoids in vitro, however, its regulation in vivo is not well established. Methods : The authors investigated the effects of glucocorticoid on the accumulation of mRNA encoding SP-B and SP-C protein content of the lung. Adult rats were given different doses of subcutaneous dexamethasone and sacrificed at 24 hours and 1 week. SP-B and SP-C mRNA were measured by a filter hybridization method. Results : 1) The accumulation of SP-B mRNA at 24 hours after 0.2 mg/kg dexamethasone treatment was increased by 23.7%. 2) The accumulation of SP-B mRNA at 1 week after 2 mg/kg dexamethasone treatment was significantly increased by 96.6%(P<0.001). 3) The accumulation of SP-C mRNA at 24 hours after 0.2 mg/kg dexamethasone treatment was significantly increased by 42.7%(P<0.01). 4) The accumulation of SP-C mRNA at 1 week after 2 mg/kg dexamethasone treatment was significantly increased by 60.0% (P<0.01). Conclusion : The authors concluded that dexamethasone treatment in vivo resulted in increased levels of SP-B mRNA and SP-C mRNA. These results suggested that dexamethasone stimulates the synthesis of hydrophobic proteins associated with surfactant.

The Modulation of Radiosensitivity by Combined Treatment of Selective COX-2 Inhibitor, NS 398 and EGF Receptor Blocker AG 1478 in HeLa Cell Line (선택적 COX-2 억제제 NS 398과 EGF 수용체 차단제 AG 1478의 복합투여가 HeLa 세포주의 방사선 감수성에 미치는 영향)

  • Youn Seon Min;Oh Young Kee;Kim Joo Heon;Park Mi Ja;Seong In Ock;Kang Kimun;Chai Gyuyong
    • Radiation Oncology Journal
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    • v.23 no.1
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    • pp.51-60
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    • 2005
  • Purpose : Selective inhibition of multiple molecular targets may improve the antitumor activity of radiation. Two specific inhibitors of selective cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) were combined with radiation on the HeLa cell line. To investigate cooperative mechanism with selective COX-2 inhibitor and EGFR blocker, in vitro experiments were done. Materials and Methods : Antitumor effect was obtained by growth inhibition and apoptosis analysis by annexin V-Flous method. Radiation modulation effects were determined by the clonogenic cell survival assay. Surviving fractions at 2 Gy ($SF_2$) and dose enhancement ratio at a surviving fraction of 0.25 were evaluated. To investigate the mechanism of the modulation of radiosensitivity, the cell cycle analyses were done by flow cytometry. The bcl-2 and bax expressions were analyzed by western blot. Results : A cooperative effect were observed on the apoptosis of the HeLa ceil line when combination of the two drugs, AG 1478 and NS 398 with radiation at the lowest doses, apoptosis of $22.70\%$ compare with combination of the one drug with radiation, apoptosis of $8.49\%$. In cell cycle analysis, accumulation of cell on $G_0/G_l$ phase and decrement of S phase fraction was observed from 24 hours to 72 hours after treatment with radiation, AG 1478 and NS 398. The combination of NS 398 and AG 1478 enhanced radiosensitivity on a concentration-dependent manner in HeLa cells with dose enhancement ratios of 3.00 and $SF_2$ of 0.12 but the combination of one drug with radiation was not enhanced radlosensitivity with dose enhancement ratios of 1.12 and SF2 of 0.68 (p=0.005). The expression levels of bcl-2 and bax were reduced when combined with AG 1478 and NS 398. Conclusion : Our results indicate that the selective COX-2 inhibitor and EGFR blocker combined with radiation have potential additive or cooperative effects on radiation treatment and may act through various mechanisms including direct inhibition of tumor cell proliferation, suppression of tumor cell cycle progression and inhibition of anti-apoptotic proteins.

Clinical Outcome of Elective Single Embryo Transfer Compared to Elective Double Embryo Transfer Performed at the Cleavage Stage (2개 대비 1개의 난할기 선발배아 이식에 따른 임상 결과)

  • Kang, Sang-Min;Lee, Sang-Won;Jeong, Hak-Jun;Chae, Soo-Jin;Yoon, San-Hyun;Lim, Jin-Ho;Lee, Seong-Goo
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.4
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    • pp.349-359
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    • 2010
  • Objective: This study was performed to compare the clinical outcome of elective single embryo transfer (eSET) performed at the cleavage stage to that of elective double embryo transfer (eDET). Methods: Of the women less than 36 years old who visited Daegu Maria from January 2008 to April 2009, the only women (n=330) with more than 8 mm of endometrial thickness and at least one good quality embryo, who were treated with GnRH agonist long protocol, were included in this study. After information about complications that can arise by multiple embryo transfer, either eSET or eDET was conducted by their request (167 and 163, respectively).Results: The implantation rate of eSET group was significantly higher than that of eDET group (53.9% vs. 40.2%, p<0.01). The twin pregnancy rate of eSET group was significantly lower than that of eDET group (1.1% vs. 32.3%, p<0.001). However, there were no significant differences between two groups in the clinical pregnancy (53.3% vs. 60.7%, p=0.172), ongoing pregnancy (47.3% vs. 54.6%, p=0.185) and live birth rates (44.9% vs. 50.9%, p=0.275). The number of the surplus embryos which developed to the blastocyst stage and cryopreserved at that stage was significantly higher in eSET group than that of eDET group ($3.2{\pm}2.6$ vs. $2.1{\pm}2.4$, p<0.001). Conclusion: These results suggest that eSET should reduce significantly the multiple baby pregnancy without decreasing the whole pregnancy rate in women with less than 36 years old.

Effects of Gamma Irradiation on Color Changes and Antioxidative Activities of Caesalpinia sappan L. (감마선 조사에 의한 소목추출물의 색상변화 및 항산화 활성 평가)

  • Kwon, Hyung-Jun;Jung, U-Hee;Park, Hae-Ran;Shin, Dong-Hwa;Jo, Sung-Kee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.1055-1061
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    • 2007
  • Caesalpinia sappan L. is an indeciduous tree distributed in China, India, Miyanmar and Vietnam. Its heartwood has long been used in oriental folk medicines to treat diseases. In this study, antioxidative activities of Caesalpinia sappan L. and the effect of gamma irradiation on its chemical and biological properties were investigated. The ethyl acetate fraction (EtOAc fr.) of Caesalpinia sappan L. was irradiated with 100 kGy of gamma ray. The dark red color of EtOAc fr. was significantly (p<0.05) removed by irradiation (Hunter L and b values increased and a value decreased). The total phenolic content of EtOAc fr. was 865 mg/g and it was increased to 1195 mg/g by gamma irradiation. DPPH radical and superoxide anion radical scavenging activities and lipid peroxidation inhibitory activity of EtOAc fr. were very high and its activities were also increased by gamma irradiation. EtOAc fr. also inhibited the irradiation-induced DNA damage of lymphocyte as determined by comet assay. In conclusion, EtOAc fr. of Caesalpinia sappan L. extract showed high antioxidative activities in vitro. Furthermore, gamma irradiation on EtOAc fr. ameliorated the color and antioxidative properties. Therefore, it can be suggested that Caesalpinia sappan L. may be a good material for antioxidant function and gamma irradiation may be applied for the improvement of chemical and biological properties of Caesalpinia sappan L.

Effects of Zizyphus jujuba var. boeunesis Extracts on the Growth of Intestinal Microflora and Its Antioxidant Activities (대추 추출물이 장내 미생물의 생육에 미치는 영향 및 항산화 활성)

  • Jeong, Hye-Mi;Kim, Yi-Seul;Ahn, Seung-Joon;Auh, Mi-Sun;Ahn, Jun-Bae;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.4
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    • pp.500-508
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    • 2011
  • This study was carried out to investigate the effects of jujube extracts on intestinal microflora, along with their antioxidant activities, according to extraction method. The antimicrobial activities of the extracts were measured using the agar diffusion method with a jujube extract concentration of 50 mg/mL. Neither the first nor second jujube extracts were inhibitory against the tested intestinal bacteria. However, water extracts of jujube significantly enhanced the growth of lactic acid bacteria, especially Bifidobacterium bifidum and Bifidobacterium adolescentis. Total phenol compounds and flavonoid compounds were higher in the 1st than in the 2nd water extracts. The EDA values of both water and ethanol extracts increased in proportion to the extract concentration. The 1st water extract showed the highest value among all the others, which was 85.60% at the concentration of 0.05 mg/mL. Furthermore, the 1st water extract showed stronger antioxidant activity than the other samples with an activity of 679.91 mg AA eq/g. These results support the potential use of jujube water extracts as a functional food component and a valuable resource for the development of nutraceutical foods, to increase the growth of Bifidobacterium spp. in the human intestine.

The Mechanisms of Resistance to TNF in TNF-Sensitive Cancer Cells Transfected with TNF-$\alpha$ Gene Using Retroviral Vector (암세포에서 Retroviral Vector를 이용한 종양괴사인자 유전자 이입후 획득된 종양괴사인자 내성의 기전)

  • Lee, Hyuk-Pyo;Oh, Yeon-Mok;Yoo, Chul-Gyu;Kim, Young-Whan;Shim, Young-Soo;Han, Sung-Koo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.547-558
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    • 1997
  • Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to hoot. In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-$\alpha$ gene transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164, a murine fibrosarcoma cell line, using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide ( translation inhibitor). Results : WEHI164 which was sensitive to TNF became resistant to TNF after being transfected with TNF-$\alpha$ gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ gene transfection may be associated with synthesis of some protective proteins.

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