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Evaluation of Antioxidant Activity of Sugar Alcohols Using TOSC (Total Oxy-radical Scavenging Capacity) Assay (TOSC 법을 이용한 당알코올의 항산화 활성 평가)

  • Kang, Keon-Wook;Kwak, Sang-Hoon;Yun, Sei-Young;Kim, Sang-Kyum
    • Toxicological Research
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    • v.23 no.2
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    • pp.143-150
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    • 2007
  • Although animal and epidemiological studies have suggested oxidative stress as an etiological factor in pathogenesis including cancer, inflammation, sepsis, fibrosis, cardiovascularlneurodegenerative diseases and aging-related disorders, conflicting results have been obtained in clinical trial with antioxidants. The reason for this discrepancy remains unknown but may be due, in part, to the lack of a validated assay system for evaluating antioxidant capacity. The antioxidant activity of a series of sugar alcohols against peroxyl radicals, hydroxyl radicals and peroxynitrites was determined by the total oxy-radical scavenging capacity (TOSC) assay and cell-based assay using H4IIE cells. Specific TOSC values calculated from the slope of the linear regression for erythritol, xylitol, sorbitol or mannitol against peroxyl radicals was $2.1{\pm}0.2,\;3.7{\pm}0.3,\;9.1{\pm}0.3$ or $8.7{\pm}1.1$ TOSC/mM, respectively. Specific TOSC values for erythritol, xylitol, sorbitol or mannitol against peroxynitrite was $1.9{\pm}0.3,\;3.9{\pm}0.4,\;7.8{\pm}0.7$ or $7.7{\pm}0.5$ TOSC/mM, respectively. These results suggest that oxy-radical scavenging capacity is dependent on the number of aliphatic hydroxyl group in sugar alcohols of monosaccharide. Tert-butylhydroperoxide (t-BHP)-induced cell toxicity determined by MTT assay was marginally attenuated by 10 mM erythritol, but completely inhibited by 10 mM xylitol, 2 mM sorbitol or 0.75 mM maltitol, a disaccharide alcohol. Oxidative stress markers, such as glutathione (GSH) and malondial-dehyde (MDA) levels, were measured in t-BHP-treated cells using HPLC equipped with a fluorescence detector and a reverse phase column. Erythritol did not change the levels of GSH and MDA in H411E cells treated with t-BHP. The t-BHP-induced changes in cellular GSH and MDA levels were ameliorated by 10 mM xylitol and completely blocked by 10 mM sorbitol and maltitol. These results indicate that sugar alcohols protect cells against oxidative stress via scavenging oxy-radical and suggest that TOSC assay in conjunction with cell-based assay is a valid method for evaluating antioxidant capacity of natural and synthetic chemicals.

Effect of Culture Media and Co-culture with Bovine and Rabbit Oviductal Epithelial Cells on In Vitro Development of Rabbit Embryos (토끼 수정란의 체외발달에 미치는 배양액 및 소와 토끼의 난관상피세포들과의 공배양 효과)

  • 노규진;이효종;송상현;윤희준;박충생
    • Korean Journal of Animal Reproduction
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    • v.18 no.1
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    • pp.39-46
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    • 1994
  • This experiment was carried out to develop an in vitro culture system for rabbit embryos. The zygotes or 2-cell embryos were collected from the oviducts of the superovulated and mated does with D-PBS/10% FCS at 24 hours after hCG injection. The in vitro developmental rate of blastocyst formation and the number of nuclei in the embryos were examined under the following treatments; 1) TCM-199 with 10% FCS, 2) EBSS with 10% FCS, 3) rabbit vitreous humor(VH), 4) TCM-199 with 10% FCS+BOEC, 5) TCM-199 with 10% FCS+ROEC, 6) EBSS with 10% FCS+BOEC and 7) EBSS with 10% FCS+ROEC. For a comparative study of in vivo and in vitro development, the fresh blastocysts, which were developed in vivo for 96 hours after hCG injection, were collected from the uterus and their numbers of nuclei were counted. 1. The zygotes or 2-cell embryos developed to the blastocyst stage in TCM-199, EBSS and VH at the rates of 93, 92 and 89%, respectively. 2. The higher developmental rates 95~98% of blastocyst formation was achieved when the embryos were co-cultured with a monolayer of bovine or rabbit oviductal epithelial cells in TCM-199 or EBSS. No significant difference in developmental rates was shown between bovine and rabbit oviductal epithelial cells. 3. In a comparative study of in vivo and in vitro development, the total numbers of nuclei were significantly less in the in vitro cultured embryos(104~224) than the in vivo developed embryos(1, 0090 at 96 hours after hCG injectin. 4. The mean cell cycle numbers in the embryos cultured for 72 hours in TCM-199 with 10% FCS, EBSS with 10% FCS, TCM-199 with 10% FCS+BOEC, TCM-199 with 10% FCS+ROEC, EBSS with 10% FCS+BOEC and in vivo was 7.38, 6.63, 7.76, 7.69, 7.01 and 9.92, respectively. From these results, it can be suggested the optimal culture system for in vitro culture of rabbit embryos is a co-culture system with bovine or rabbit oviductal epithelial cells in TCM-199 with 10% FCS. Considering the significant reduction in total numbers of nuclei in the in vitro cultured embryos, the advanced research on development of in vitro culture system for rabbit embryos is expected.

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Effects of dietary glutamine and arginine supplementation on performance, intestinal morphology and ascites mortality in broiler chickens reared under cold environment

  • Abdulkarimi, Rahim;Shahir, Mohammad Hossein;Daneshyar, Mohsen
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.1
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    • pp.110-117
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    • 2019
  • Objective: An experiment was conducted to evaluate the effects of dietary glutamine (Gln) and arginine (Arg) supplementation on performance, intestinal morphology and ascites mortality in broilers. Methods: A total of 675 day old chicks were randomly allocated to 9 experimental groups in a $3{\times}3$ factorial arrangement based on a completely randomized design with 5 replicates of 15 chicks. Three levels of dietary Gln (0%, 0.5%, and 1%) and Arg (100%, 130%, and 160% of Ross recommendation) supplementation were used in ascites inducing condition ($15^{\circ}C{\pm}1^{\circ}C$) from 7 to 42 days of age. Results: Dietary supplementation of Gln increased body weight gain during grower, finisher and total periods (p<0.05) and increased feed intake during total period. Ascites mortality was decreased by Gln supplementation (p<0.05). Gln supplementation increased the villus height (VH) and crypt depth (CD) in duodenum and jejunum (p<0.05). Arg supplementation decreased CD in duodenum and jejunum, and increased ileum villus width (VW) and also VH/CD ratio in duodenum and jejunum (p<0.05). Both Gln and Arg increased the goblet cell number (GCN) in duodenum whereas Gln supplementation decreased GCN in jejunum and ileum (p<0.05). The $Gln{\times}Arg$ interaction were observed for right ventricle (RV)/total ventricular (TV) ratio, VH, VW, CD, VH/CD. Conclusion: It was concluded that dietary 0.5% Gln alone or along with 130% Arg of Ross requirement, improve the intestinal morphology and performance and hence decrease the ascites mortality in broiler chickens with cold induced ascites.

Research on Immune Enhancing Effect and Safety of Wasong (Orostachys japonicus) Extract: Study Protocol for a Single Center, Randomized, Double-blind, Placebo-controlled, Clinical Trial (와송 추출물의 면역기능 개선 효과 및 안전성 연구: 단일기관, 무작위배정, 이중눈가림, 위약대조 비교, 임상연구 프로토콜)

  • Choi, Jin Yong;Choi, Jun Yong;Lim, Hyun Woo;Kim, Jeong;Kim, So Yeon;Han, Chang Woo
    • Herbal Formula Science
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    • v.25 no.2
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    • pp.135-143
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    • 2017
  • Objectives : This trial aimed to determine if Wasong (Orostachys japonicus) extract can enhance immune system and is safe enough to be approved as a health functional food. Methods : Total 62 people, aged 45 and older, will be recruited to participate in a randomized, double-blind, placebo-controlled clinical trial. This study will compare Wasong extract and placebo. Wasong group will take 1g of Wasong extract, once a day, for 8 weeks. Placebo group will take 1g of crystalline cellulose as placebo, once a day, for 8 weeks. Outcomes will be measured at the baseline, the end of 4th week, and 8th week. Primary outcomes are the ratio of NK cells/total lymphocytes and the ratio of T-helper cells/T-suppressor cells. Secondary outcomes are total white blood cell count, the ratio of neutrophils, lymphocytes, and monocytes in total leukocytes, the ratio of total T cells, T-helper cells, T-suppressor cells, and B cells to lymphocytes, the amount of blood IgM, IgG, IgA, and cytomegalovirus (CMV) IgG, and blood metabolite target &global analysis. Results : This trial was approved by institutional review board of Pusan National University Korean Medicine Hospital (registry number: 2016006), and registered in Clinical Research information Service, one of WHO International Clinical Trials Registry Platform (registry number: PRE20161006-002). Recruitment opened in February 2017 and is supposed to be completed by August 2017. The result is expected to be published by June 2018. Conclusion : This trial will provide clinical information to determine the efficacy and safety of Wasong in enhancing immune system of middle-aged and older people.

Fed-batch Fermentations of Recombinant Escherichia coli to Produce Bacillus macerans CGTase

  • Park, Yong-Cheol;Kim, Chang-Sup;Kim, Chung-Im;Choi, Kyu-Hwan;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.323-328
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    • 1997
  • The recombinant Escherichia coli BL21(DE3)pLysE : pTCGT1 was grown to overproduce Bacillus macerans cyclodextrin glucanotransferase (CGTase) able to synthesize ${\alpha}$-cyclodextrin (CD) with a selectivity of 67%. A number of batch fermentations were performed to test the possibility of using lactose as an inducer of the E. coli T7 promoter system. A mixture of isopropyl ${\beta}$-D-thiogalactoside (IPTG) and lactose (1 : 1) gave a maximum CGTase activity of 2.4 U/ml, which was higher than the value obtained with induction by IPTG alone. Fed-batch fermentations involving a glucose-controlled growth period followed by a gene-expression phase with mixtures of IPTG and lactose were employed to achieve high cell density and thereby increase total CGTase activity. Optimized fed-batch fermentation using the modified inducer (IPTG : lactose=1 : 3) and 100 g/l yeast extract solution in the gene-expression phase resulted in a maximum CGTase activity of 62.9 U/ml and a final cell mass of 53.5 g/l, corresponding to a 31-fold increase in CGTase activity and a 29-fold increase in cell mass compared with the control batch fermentation.

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Design Optimization of a 500W Fuel Cell Stack Weight for Small Robot Applications (소형로봇용 500W급 연료전지 스택무게 최적화 설계)

  • Hwang, S.W.;Choi, G.H.;Park, Sam.;Ench, R. Michael;Bates, Alex M.;Lee, S.C.;Kwon, O.S.;Lee, D.H.
    • Journal of the Korean Solar Energy Society
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    • v.32 no.spc3
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    • pp.275-281
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    • 2012
  • Proton Exchange Membrane Fuel Cells (PEMFC) are the most appropriate for energy source of small robot applications. PEMFC has superior in power density and thermodynamic efficiency as compared with the Direct Methaol Fuel Cell (DMFC). Furthermore, PEMFC has lighter weight and smaller size than DMFC which are very important factors as small robot power system. The most significant factor of mobile robots is weight which relates closely with energy consumption and robot operation. This research tried to find optimum specifications in terms of type, number of cell, active area, cooling method, weight, and size. In order to find optimum 500W PEMFC, six options are designed in this paper and studied to reduce total stack weight by applying new materials and design innovations. However, still remaining problems are thermal management, robot space for energy sources, and soon. For a thermal management, design options need to analysis of Computational Fluid Dynamics (CFD) for determining which option has the improved performance and durability.

Immunohistochemical evaluation of p63 and cyclin D1 in oral squamous cell carcinoma and leukoplakia

  • Patel, Sunit B.;Manjunatha, Bhari S.;Shah, Vandana;Soni, Nishit;Sutariya, Rakesh
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.43 no.5
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    • pp.324-330
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    • 2017
  • Objectives: There are only a limited number of studies on cyclin D1 and p63 expression in oral squamous cell carcinoma (OSCC) and leukoplakia. This study compared cyclin D1 and p63 expression in leukoplakia and OSCC to investigate the possible correlation of both markers with grade of dysplasia and histological grade of OSCC. Materials and Methods: The study included a total of 60 cases, of which 30 were diagnosed with OSCC and 30 with leukoplakia, that were evaluated immunohistochemically for p63 and cyclin D1 expression. Protein expression was correlated based on grades of dysplasia and OSCC. Results: Out of 30 cases of OSCC, 23 cases (76.7%) were cyclin D1 positive and 30 cases (100%) were p63 positive. Out of 30 cases of leukoplakia, 21 cases (70.0%) were cyclin D1 positive and 30 (100%) were p63 positive (P<0.05). Conclusion: The overall expression of cyclin D1 and p63 correlated with tumor differentiation, and increases were correlated with poor histological grades, from well-differentiated to poorly-differentiated SCC. Increased cyclin D1 and p63 expression was associated with the severity of leukoplakia. Based on these results cyclin D1 and p63 products can be a useful tool for improved leukoplakia prognosis.

Late Stage and Grave Prognosis of Esophageal Cancer in Thailand

  • Nun-anan, Pongjarat;Vilaichone, Ratha-korn
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.1747-1749
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    • 2015
  • Background: Esophageal cancer is one of the major health concerns in Southeast Asian countries, including Thailand. However, only a limited number of studies have been reported from this region. This study was designed to evaluate the prevalence, clinical characteristics and survival rate of esophageal cancer in Thailand. Materials and Methods: Clinical information, histological features and endoscopic findings were collected from a tertiary care center in central region of Thailand between September 2011- November 2014 and reviewed. Results: A total of 64 esophageal cancer patients including 58 men and 6 women with mean age of 62.6 years were enrolled. Common presenting symptoms were dysphagia (74%), dyspepsia (10%) and hematemesis (8%). Mean duration of symptoms prior to diagnosis was 72 days. Esophageal stenosis with contact bleeding was the most common endoscopic finding (55.6%). The location of cancer was found in proximal (16%), middle (50%) and distal (34%) esophagus. Squamous cell carcinoma was far more common histology than adenocarcinoma (84.2% vs 10.5%). However, esophageal adenocarcinoma was significantly more common than squamous cell carcinoma in distal area of esophagus (100% vs 22.9%; p=0.0001, OR=1.6, 95%CI=1.1-2.2). Esophageal cancer stages 3 and 4 accounted for 35.2% and 59.3% respectively. Overall 2-year survival rate was 20% and only 16% in metastatic patients. Conclusions: Most esophageal cancer patients in Thailand have squamous cell carcinoma and nearly all present at advanced stage with a grave prognosis. Screening of high risk individuals and early detection might be important keys to improve the survival rate and treatment outcome in Thailand.

Long-Term Starvation Induces the Viable-but-Nonculturable Condition in Lactobacillus crispatus KLB46

  • Lee, Seok-Yong;Kim, Ju-Hyeon;Jang, Jeong-Eun;Kim, Seung-Cheol;Yun, Hyeon-Sik;So, Jae-Seong
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.918-922
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    • 2001
  • In a previous study, we have isolated a number of lactobacilli from Korean women, and one of them (KLB46) was identified as Lactobacillus crispatus by 16S rRNA gene sequencing. For the ecological treatment of bacterial vaginosis (BV) cell suspension of L. crispatus KLB46 was instillated into BV patients. L. crispatus KLB46 was found to persist for several days in cell suspension with no nutrients. In this study, in order to assess the influence of starvation on physiological activity, we compared the viability and culturability of KLB46 following suspension in various buffer solutions. A pair of in situ fluorescent dye was used to assess viability (i.e. membrane integrity) and the culturability was examined by plate count assay. A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit $(BacLight^{TM})$ was applied to estimate both viable and total counts of bacteria in cell suspension. $BacLight^{TM}$ is composed of two nucleic acid-binding stains ($SYTO\;9^{TM}$ and propidium iodide). $SYTO\;9^{TM}$ penetrates all bacterial membranes and stains the cells green while propidium iodide only penetrates cells with damaged membranes, therefore the combination of the two stains produces red fluorescing cells. Optimal staining conditions for $BacLight^{TM}$ were found to be with 0.0835M $SYTO\;9^{TM}$ and 0.05M propidium iodide for 15 min incubation at room temperature in dark. When cells were microscopically examined during 140 hours of starvation, the culturability decreased markedly while the viability remained relatively constant, which suggests that large fraction of KLB46 cells became viable but non-culturable (VBNC) upon starvation.

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Effects of Tubulyzines, Novel Microtubule-Binding Triazine Molecules, on Endothelial Progenitor Cell Differentiation

  • Park, Hyo-Eun;Lee, Soo-Young;Ahn, Hyun-Young;Shin, Jong-Cheol;Chang, Young-Tae;Joe, Young-Ae
    • Biomolecules & Therapeutics
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    • v.11 no.2
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    • pp.85-90
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    • 2003
  • Microtubule-binding molecules have been developed as anti-cancer agents to overcome the toxicities of current chemotherapeutics and also have potential for use as anti-angiogenic agents. In this work, we examined the effect of novel triazine compounds, Tubulyzines (microTUBUle LYsing triaZINE), derived from the orthogonal synthesis of a triazine library, on endothelial progenitor cell differentiation. When mononuclear cells isolated from human cord blood were cultured on fibronectin-coated plates for 7 days, all the Tubulyzine compounds A, B, and C (TA, TB, and TC) tested decreased the number of adherent cells in a dose-dependent manner in a coo. centration ranges of 2-5 to $80\mu\textrm{M}$. TA ($IC_{50}$=$20\mu\textrm{M}$) showed slightly more potent activity than TB and TC. Adherent cells treated with TA also exhibited a lower level of ability to ac-LDL uptake, with low ratios of positive cells out of total adherent cells, in a dose-dependent manner and weak expression of endothelial lineage markers, KDR, CD31, and vWF at $20\mu\textrm{M}$. Therefore, these results suggest that tubulyzine A (TA) can be effectively used for the inhibition of new vessel growth by inhibiting differentiation of endothelial progenitor cells.