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Differential Effect of MyD88 Signal in Donor T Cells on Graft-versus-Leukemia Effect and Graft-versus-Host Disease after Experimental Allogeneic Stem Cell Transplantation

  • Lim, Ji-Young;Ryu, Da-Bin;Lee, Sung-Eun;Park, Gyeongsin;Choi, Eun Young;Min, Chang-Ki
    • Molecules and Cells
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    • v.38 no.11
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    • pp.966-974
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    • 2015
  • Despite the presence of toll like receptor (TLR) expression in conventional $TCR{\alpha}{\beta}$ T cells, the direct role of TLR signaling via myeloid differentiation factor 88 (MyD88) within T lymphocytes on graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect after allogeneic stem cell transplantation (allo-SCT) remains unknown. In the allo-SCT model of C57BL/6 ($H-2^b$) ${\rightarrow}$ B6D2F1 ($H-2^{b/d}$), recipients received transplants of wild type (WT) T-cell-depleted (TCD) bone marrow (BM) and splenic T cells from either WT or MyD88 deficient (MyD88KO) donors. Host-type ($H-2^d$) P815 mastocytoma or L1210 leukemia cells were injected either subcutaneously or intravenously to generate a GVHD/GVL model. Allogeneic recipients of MyD88KO T cells demonstrated a greater tumor growth without attenuation of GVHD severity. Moreover, GVHD-induced GVL effect, caused by increasing the conditioning intensity was also not observed in the recipients of MyD88KO T cells. In vitro, the absence of MyD88 in T cells resulted in defective cytolytic activity to tumor targets with reduced ability to produce IFN-${\gamma}$ or granzyme B, which are known to critical for the GVL effect. However, donor T cell expansion with effector and memory T-cell differentiation were more enhanced in GVHD hosts of MyD88KO T cells. Recipients of MyD88KO T cells experienced greater expansion of Foxp3- and IL4-expressing T cells with reduced INF-${\gamma}$ producing T cells in the spleen and tumor-draining lymph nodes early after transplantation. Taken together, these results highlight a differential role for MyD88 deficiency on donor T-cells, with decreased GVL effect without attenuation of the GVHD severity after experimental allo-SCT.

The Effects of Syzygium aromaticum extract Spread on the Allergic Contact Dermatitis induced by DNCB (정향 추출물 도포가 DNCB로 유발된 알레르기성 접촉 피부염에 미치는 영향)

  • Lee, Kyung-Yeob;Kang, Da-Hae;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.26 no.4
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    • pp.1-14
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    • 2013
  • Objectives : The purpose of this study is to investigate effects of Syzygium aromaticum (SAE) spread on the allergic contact dermatitis caused by 2,4-dinitro-chlorobezene (DNCB). Methods : Forty-two mice were divided into six groups ; normal, negative control (DNCB-treated), positive control (DNCB + 1% pimecrolimus), experimental group I, II and III. control and experimental groups were induced allergic contact dermatitis by DNCB. Experimental group I(DNCB + 0.2% SAE), II(DNCB + 1% SAE) and III(DNCB + 5% SAE) were spread SAE and positive control was spread the 1% pimecrolimus. In this study, effect of SAE on clinical aspects on the skin, histopathological change, the blood level of IgE, cytokines, histamine were investigated. In addition, effect of SAE on spleen $CD4^+/CD8^+$ T cell subset was investigated. Results : 1. In experimental group I, II and III, erythemas and edema were more reduced than negative control. 2. In experimental group I, II and III inflammatory edema and the numbers of infiltrated inflammatory cells were more reduced than negative control. 3. In experimental group I, II and III, clinical skin score was more reduced than negative control. 4. In experimental group II and III, the thickness of skin was statistically significant reduced than negative control. 5. In experimental group II and III, histamine release was statistically significant reduced than negative control in dose-dependantly. 6. In experimental group II and III, cytokines (IL-1${\beta}$, TNF-${\alpha}$, IL-4, IL-6, IL-10) were statistically significant reduced than negative control in dose-dependantly. 7. In experimental group I, II and III, the level of total IgE was statistically significant reduced than negative control in dose-dependantly. 8. In experimental group III, $CD4^+$ and $CD8^+$ T cells were statistically significant decreased similar to the positive control. Conclusions : According to above experiments, Syzygium aromaticum(SAE) was effective on allergic contact dermatitis.

Establishment of a Simple and Effective Method for Isolating Male Germline Stem Cells (GSCs) from Testicular Cells of Neonatal and Adult Mice

  • Kim Kye-Seong;Lim Jung-Jin;Yang Yun-Hee;Kim Soo-Kyoung;Yoon Tae-Ki;Cha Kwang-Yul;Lee Dong-Ryul
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1347-1354
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    • 2006
  • The aims of this study were to establish a simple and effective method for isolating male germline stem cells (GSCs), and to test the possibility of using these cells as a new approach for male infertility treatment. Testes obtained from neonatal and adult mice were manually decapsulated. GSCs were collected from seminiferous tubules by a two-step enzyme digestion method and plated on gelatin-coated dishes. Over 5-7 days of culture, GSCs obtained from neonates and adults gave rise to large multicellular colonies that were subsequently grown for 10 passages. During in vitro proliferation, oct-4 and two immunological markers (Integrin ${\beta}1,\;{\alpha}6$) for GSCs were highly expressed in the cell colonies. During another culture period of 6 weeks to differentiate to later stage germ cells, the expression of oct-4 mRNA decreased in GSCs and Sertoli cells encapsulated with calcium alginate, but the expression of c-kit and testis-specific histone protein 2B(TH2B) mRNA as well as the localization of c-kit protein was increased. Expression of transition protein (TP-l) and localization of peanut agglutinin were not seen until 3 weeks after culturing, and appeared by 6 weeks of culture. The putative spermatids derived from GSCs supported embryonic development up to the blastocyst stage with normal chromosomal ploidy after chemical activation. Thus, GSCs isolated from neonatal and adult mouse testes were able to be maintained and proliferated in our simple culture conditions. These GSCs have the potential to differentiate into haploid germ cells during another long-term culture.

Bioequivalence of Cadilan Tablet 12.5 mg to Dilatrend® Tablet 12.5 mg (Carvedilol 12.5 mg) (딜라트렌 정 12.5밀리그람(카르베딜롤 12.5밀리그람)에 대한 카딜란 정 12.5밀리그람의 생물학적동등성)

  • Kim, Se-Mi;Shin, Sae-Byeok;Kim, Ju-Hwan;Kwon, In-Ho;Kim, Yong-Hee;Lee, Sang-No;Cho, Hea-Young;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.38 no.6
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    • pp.413-419
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    • 2008
  • Carvedilol, is a nonselective $\beta$-blocking agent and it also has vasodilating properties that are attributed mainly to its blocking activity at ${\alpha}_1$-receptors. The purpose of the present study was to evaluate the bioequivalence of two carvedilol tablets, $Dilatrend^{(R)}$ tablet 12.5 mg (Chong Kun Dang Pharmaceutical Co., Ltd.) and Cadilan tablet 12.5 mg (KyungDong Pharmaceutical. Co., Ltd.), according to the guidelines of the Korea Food and Drug Administration (KFDA). The release of carvedilol from the two carvedilol formulations in vitro was tested using KP VIII Apparatus II method with pH 4.5 dissolution medium. Thirty two healthy male subjects, $25.00{\pm}3.09$ years in age and $70.71{\pm}11.35\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After a single tablet containing 12.5 mg as carvedilol was orally administered, blood samples were taken at predetermined time intervals and the concentrations of carvedilol in serum were determined using HPLC with fluorescence detector. The dissolution profiles of two formulations were similar in the tested dissolution medium. The pharmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$, $C_{max}$ and untransformed $T_{max}$. The results showed that the differences between two formulations based on the reference drug, $Dilatrend^{(R)}$ tablet 12.5 mg, were 4.66%, 8.33% and -7.45% for $AUC_t$, $C_{max}$ and $T_{max}$, respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log 0.8 to log 1.25 (e.g., $\log\;0.9823{\sim}\log\;1.1042$ and $\log\;1.0132{\sim}\log\;1.1875$ for $AUC_t$ and $C_{max}$, respectively). Thus, the criteria of the KFDA bioequivalence guideline were satisfied, indicating Cadilan tablet 12.5 mg was bioequivalent to $Dilatrend^{(R)}$ tablet 12.5 mg.

The Effects of Cholates on Smooth Muscle Strips and Cardiac Muscle (담즙산(膽汁酸)이 수종평골근(數種平滑筋) 및 심근표본(心筋標本)에 미치는 영향(影響))

  • Ro, J.Y.;Cho, T.S.;Hong, S.S.
    • The Korean Journal of Pharmacology
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    • v.16 no.1 s.26
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    • pp.41-50
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    • 1980
  • Chenodeoxycholic acid(CDCA) has been used as a gallstone dissolving agent since 1972. Recently, ursodeoxycholic acid(UDCA) has been reported to be effective in dissolving gallstones. Both bile acids increased bile flow. The increase in bile flow associated with an increase in cholesterol level in bile after CDCA or UDCA infusion was reported. In this study, using the smooth muscle strips of guinea pig and fowl, responses of the cholates were observed. In addition, the influence of adrenergic blocking agents on the response of the strips to cholates was investigated. Also the effects of cholates on cardiac function were examined by using isolated atria of rabbit and heart of anesthetized frog. The results are as follows: 1) All cholates, such as UDCA, CDCA, and CA produced a marked inhibitory effect on the motility in isolated duodenal strip of guinea pig and fowl, however, only UDCA showed the contraction in the isolated esophagus of fowl. These effects of cholates were blocked by propranolol. 2) In isolated guinea pig stomach strip and gall bladder, cholates exhibited a marked inhibitory effect on the motility and the effects due to UDCA and CA were blocked by phenoxybenzamine while CDCA was not affected. 3) The spontaneous and ouabain induced arrhythmia was partially abolished by cholates. However, concomitant administration of cholates with ouabain or epinephrine caused a marked prolongation in occurrence of atrial arrhythmia in comparison with ouabain or epinephrine alone in isolated rabbit atria. 4) In the heart of anesthetized frog, the epinephrine-induced arrhythmia was partially abolished by cholates. The combined treatment with cholates and ouabain or epinephrine produced a marked prolongation in occurrence of the arrhythmia in comparison with, ouabain or epinephrine alone. From the above results, it can be suggested that the effects of cholates on the smooth muscle of duodenum and esophagus are produced in response to adrenergic ${\beta}$-receptor and the effect or gall bladder and stomach is more likely due to the direct effect on the muscle. In addition, cholates exhibit a slight antiarrhythmic effect on heart, therefore, cholates can be classified as a nonselective antiarrhythmic drug, such as propranolol.

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Purification and Characterization of a Thermostable Xylanase from Paenibacillus sp. NF1 and its Application in Xylooligosaccharides Production

  • Zheng, Hong-Chen;Sun, Ming-Zhe;Meng, Ling-Cai;Pei, Hai-Sheng;Zhang, Xiu-Qing;Yan, Zheng;Zeng, Wen-Hui;Zhang, Jing-Sheng;Hu, Jin-Rong;Lu, Fu-Ping;Sun, Jun-She
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.489-496
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    • 2014
  • High levels of extracellular xylanase activity (211.79 IU/mg) produced by Paenibacillus sp. NF1 were detected when it was submerged-cultured. After three consecutive purification steps using Octyl-Sepharose, Sephadex G75, and Q-Sepharose columns, a thermostable xylanase (XynNF) was purified to homogeneity and showed a molecular mass of 37 kDa according to SDS-PAGE. The specific activity of the purified XynNF was up to 3,081.05 IU/mg with a 14.55-fold purification. The activity of XynNF was stimulated by $Ca^{2+}$, $Ba^{2+}$, DTT, and ${\beta}$-mercaptoethanol, but was inhibited by $Fe^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $Cu^{2+}$, SDS, and EDTA. The purified XynNF displayed a greater affinity for oat spelt xylan with the maximal enzymatic activity at $60^{\circ}C$ and pH 6.0. XynNF, which was shown to be cellulose-free, with high stability at high temperature ($70^{\circ}C-80^{\circ}C$) and low pH range (pH 4.0-7.0), is potentially valuable for various industrial applications. The enzyme hydrolyzed oat spelt xylan to yield mainly xylooligosaccharides (95.8%) of 2-4 degree of polymerization (DP2-4). Moreover, the majority of the xylooligosacharides (DP2-4) products was xylobiose (61.5%). The thermostable xylanase (XynNF) thus seems potentially usefull in the production of xylooligosaccharides.

The Nitrite Scavenging and Electron Donating Ability of Phenolic Compounds (페놀성 화합물의 아질산염 소거 및 전자공여 작용)

  • Kang, Yoon-Han;Park, Yong-Kon;Lee, Gee-Dong
    • Korean Journal of Food Science and Technology
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    • v.28 no.2
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    • pp.232-239
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    • 1996
  • Phenolic compounds are known to inhibit the nitrosation or oxidation reaction. In the present work, the effects of phenolic compounds including phenolic acids and flavonoids on the nitrite-scavenging and electron donating ability were tested as scavenger of nitrite which is believed to participate in the formation of N-nitroso compounds and investigated as electron donator. The nitrite scavenging ability appeared in all the phenolic acids and showed the highest value at PH 1.2. Among the Phenolic compounds, phenolic acids showed higher nitrite-scavenging action than some flavonoids. Futhermore, the nitrite scavenging action of phenolic compounds was pH dependent highest at pH 1.2 and lowest at pH 6.0. The electron donating ability (EDA) by reduction of ${\alpha},{\alpha}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) among hydroxybenzoic acids was in the decreasing order of gallic acid, gentisic acid, syringic acid, protocatechuic acid, salicylic acid, vanillic acid, benzoic acid and p-hydroxybenzoic acid. EDA of hydroxycinnamic acids was in the decreasing order of hydrocaffeic acid, caffeic acid, ferulic acid, p-coumaric acid and trans-cinnamic acid. EDA of flavonoids was in the decreasing order of (+)catechin, rutin, quercetin, naringin and hesperidin. Other phenolic compounds were significantly high in electron donating abilities.

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한국산 장뇌산삼의 부위별 유용성분함량 및 추출용매조건의 영향

  • 김준한;문혜경;강우원;김종국
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.10a
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    • pp.150.2-151
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    • 2003
  • 한국산 장뇌산삼의 열매, 잎, 줄기 및 뿌리를 -7$0^{\circ}C$ 동결건조 분말화시킨 시료에 대한 추출용매와 농도에 따른 유용성분의 함량을 비교, 분석하였다. 추출액의 당도는 잎과 줄기의 80% 에탄올추출액이 각각 22.58%와 22.53%로 가장 높았고, pH는 4.43-7.41 수준이었고, 갈변도는 흡광도가 잎과 뿌리 80% 에탄올추출액이 1.803과 1.085로 가장 높았다. 색도의 경우 L값는 줄기 100% 증류수추출액이 24.56, 열매 80% 메탄올추출액이 24.35로 가장 높았고, 뿌리와 잎 80% 에탄올추출액이 각각 17.47과 19.59로 가장 낮은 값을 나타내었다. a값은 잎100% 증류수추출액이 0.41로 가장 높았고, 줄기 80%메탄올추출 액이 -0.49로 가장 낮은 값을 보였다. b값은 줄기 80% 메탄올추출액이 3.69로 가장 높았고, 열매 100% 증류수추출액 이 0.45로 가장 낮은 값이었다. 주요 유리당은 sucrose, glucose 및 fructose 이었고, 유리당 총함량은 열매와 줄기 100% 증류수추출액이 각각 6733 mg/100g과 6142 mg/100g으로 가장 많은 량을 함유하였고, sucrose는 뿌리 80% 메탄올추출액 이 3673 mg/100g으로, glucose 및 fructose는 줄기 80% 에탄올추출액과 잎 80% 메탄올추출액에 각각 4283 mg/100g과 1897 mg/100g으로 높은 함유량을 나타내었다. 또한 잎과 줄기에는 xylose가 304-524 mg/100g 수준으로 함유되어 있었고, 뿌리에는 maltose가 소량 함유하고 있었다. 주요 유기산으로는 citric acid, tartaric acid 및 malic acid가 확인되었고, citric acid는 뿌리 80% 메탄올추출액이 1849 mg/100g으로 가장 높은 함량이었고, tartaric acid는 잎 80% 메탄올추출액이 3263 mg/100g으로 가장 높은 함량이었고, malic acid는 뿌리 80% 메탄올추출액이 1856 mg/100g으로 가장 높은 함량이었으며, 뿌리 80% 메탄올추출액에는 succinic acid, malonic acid 및 oxalic acid 등이 확인되었다. 유리아미노산은 L-Arginine, ${\gamma}$-Amino-n-butyric acid, Ethanolamine, L-Proline, $\beta$-Alanine 및 L-sarcosine 등 총 35종이 확인되었고, L-Arginine은 251-7379 mg/100g 수준으로 특히, 뿌리 80% 메탄올추출액에는 1319 mg/100g으로 총함유량의 79.13%로 매우 높은 함유량을 나타내었다. P, K, Na, Ca 및 Mg 등이 주된 무기질로 확인되었고, 그 중 P는 줄기 100% 증류수추출액에 15563 mg/100g으로 가장 높았고, K은 잎 80% 메탄을 추출액에 4952 mg/100g, Ca과 Na은 잎과 열매 100% 증류수추출액에 각각 3052 mg/100g과 1798 mg/100g, Mg은 잎 100% 증류수추출액에 950 mg/100g으로 매우 높은 함유량을 보였고, 또한 미량원소로는 Zn, Cu, Cr, Mn, Co, Mo, Fe 등이 함유되어 있었다.

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Effects of Lactose and Yeast on the Changes of Oligosaccharides during the Fermentation of Soy Yogurts (Lactose와 효모의 첨가가 대두요구르트 발효 중 올리고당의 변화에 미치는 영향)

  • Park, Mi-Jung;Lee, Sook-Young
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.539-545
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    • 1997
  • Lactobacillus bulgaricus and Kluyveromyces lactis were inoculated to Jangyeob and Jinpum soy milks together after the addition of different amounts of lactose to increase the contents of oligosaccharides, which were compared with single cultured samples. The contents of stachyose, raffinose, sucrose, and glucose of samples without lactose decreased by single culture method, but the oligosaccharides decreased less than in single cultured samples containing of lactose. The oligosaccharides of single cultured samples were equal or decreased compared with soy milks. While those of mixed cultured Jangyeob and Jinpum samples containing 2% lactose for 24 hr incubation increased 125.0% and 118.1%, respectively and those of samples for 36 hr incubation increased 127.0% and 141.0%, respectively, those of mixed cultured samples containing 4% lactose for 24 hr incubation increased 112.5% and 123.0%, respectively and those of samples for 36 hr incubation increased 120% and 135.9%, respectively. Therefore, the oligosaccharides in samples containing 2% lactose were slightly more than in samples containing 4% lactose. Among the cultured methods, oligosaccharides were produced in the largest amounts by the mixed culture for 36 hr. The addition of lactose in soy milks for soy yogurts was effective in the formation of oligosaccharides since the galactose, produced by the hydrolysis of lactose, was thought to be combined with sucrose by the action of ${\beta}-galactosidase$ in yeast.

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Reduction of Mouse Body Fats by Water Extract of Pleurous Ostreatus

  • Kim, Seck -Jong;Park, Cherl -Woo;Kim, Jeong-Ok;Kim, Jong -Man;Ha, Yeong-Lae
    • Preventive Nutrition and Food Science
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    • v.4 no.2
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    • pp.130-133
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    • 1999
  • Body fat-reducing ability of oyster mushroom (Pleurotus ostreatus) water extract (OMWE) was investigated of mice by supplying it drinking water. OMWE(2.95% solid content ) was prepared by extracting a low grade of the mushroom at 12$0^{\circ}C$ for 10min. The solid material of OMWE was composed of 65.2% reducing sugar, 0.23% crude fat, 0.5%total protein, 1.2% ash and 32.9% others. OMWE was appropriately diluted with drinking water. Seventy two male ICR mice(25$\pm$1 g, 7~8 weeks of age, 6 mice/cage, 18 mice/treatment) housed in polycarbonate cages containing $\beta$-chips were adopted in a temperature-and humidity-controlled facility with free access to water and diet. One week later, the mice were subjected to one of the treatments for 36days : 0 (control), 10, 50 and 100% OMWE. Drinking wter with or without OMWE was supplied twice (40ml each, 80ml in total ) daily per cage. Body weight and fed intake were recorded every three days. At the end of the experiment, mice were sacrificed to determine the chemical composition (fat, protein, ash and water). Body weight of mice treated with OMWE (10, 50 and 100%) at day 36 was 35.9, 35.9and 35.5g per mouse , respectively, and not significantly reduced as compared to that (36.5g/mouse) of control mice. Average body fat of 0,10,50 and 10% OMWE -treated mice was 14.3, 13.1, 10.7 and 12.0% , respectively. Body fat reduction by 50% OMWE treatment was 25.2% (p<0.05) relative to control. OMWE did not affect feed intake. The contents of body protein and ash were increased with respect to body fat decrease, while water content was not changed much. These results suggest that OMWE could reduce body fat of the mice without body weight change, giving the best effect by 50% OMWE.

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