• Title/Summary/Keyword: bio assay

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Caspase-3 Activation is Associated with Granulosa Cell Apoptosis during Follicular Atresia in Porcine Ovary (돼지 폐쇄난포내 과립세포의 자연세포사 시 캐스파제-3의 활성화)

  • Kim, Jong-Min;Chung, Jin-Yong;Kim, Ji-Young;Oh, Seung-Hoon;Song, Kang-Won;Do, Byoung-Rok;Kim, Sang-Soo;Jung, Jin;Lee, Chang-Joo;Yoon, Yong-Dal
    • Development and Reproduction
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    • v.10 no.1
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    • pp.1-7
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    • 2006
  • Ovarian follicular atresia in mammals is finely regulated by gonadotropins and sex steroid hormones. It is well known that granulosa cell pyknosis is a common cytological feature of atretic follicles in the ovary. The present study hypothesized that granulosa cell pyknosis during follicular atresia might be related to apoptotic process and associated with caspase-3 activation. Healthy (normal) and atretic follicles were isolated from porcine ovaries based on macro-morphological criteria. Isolated follicles were either processed for histological observation or used for collection of granulosa cells by aspiration. Hoechst 33258 staining of the cells showed a significantly higher number of fragmented nuclei, a typical morphological feature of apoptotic cell, in granulosa cells from atretic follicles than those from healthy follicles. In addition, the rate of cell death was significantly higher in granulosa cells from atretic follicles than healthy follicles, as measured by flow-cytometric cell cycle analysis. In situ detection of apoptotic cells by TUNEL revealed that apoptosis was mostly restricted to granulosa cells in follicles. Theca cells were TUNEL-negative. Finally, it has been shown by caspase-3 activity assay that granulosa cells from atretic follicles retain a higher caspase-3 activity compared to healthy follicles. Taken together, it is suggested that granulosa cell degeneration during folliclar atresia occurs by caspase-3-dependent apoptotic fashion.

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Effect of Continuous Feeding of Probiotics on Growth Performance, Nutrient Digestibility, Blood Urea Nitrogen and Immune Responses in Pigs (지속적인 생균제의 첨가가 돼지의 성장, 영양소 이용율, 혈중 요소태 질소 및 면역능력에 미치는 영향)

  • Kil, D.Y.;Lim, J.S.;Tian, J.Z.;Kim, B.G.;Kim, K.S.;Kim, Y.Y.
    • Journal of Animal Science and Technology
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    • v.46 no.1
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    • pp.39-48
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    • 2004
  • This experiment was conducted to investigate the effect of continuous feeding of probiotics on growth performance, nutrient digestibility, blood urea nitrogen(BUN) and immune responses in pigs. Treatments were 1) Control(basal diet), 2) P-O.l(basal diet + 0.1% probiotics) and 3) P-0.2(basal diet + 0.2% probiotics). In growth trial, a total of sixty pigs(6.17 $\pm$ 0.45 kg average body weight) weaned at 21 days of age were used. All pigs were assigned according to sex and body weight, and each treatment had 5 replicates of 4 pigs per pen in a randomized complete block(RCB) design. During 0${\sim}$8 weeks, there was no significant difference in average daily gain(ADG), average daily feed intake(ADFI) and gain:feed ratio(GfF) among treatments. During 9 - 20 weeks, ADG was improved significantly in pigs fed P-O.I or P-0.2 diets when compared to the pig fed control diet(P <0.05), but there was no significant difference in ADFI and GfF ratio. During overall period, ADG, ADFI and GfF ratio were not significantly different among treatments. In the first metabolic trial(17.93 $\pm$1.45kg average body weight), apparent digestibility of OM, protein, fat in pigs fed P-O.l and P-0.2 diets were greater than in pigs fed control diet(P <0.05) and ash digestibility in pigs fed P-0.2 diet was significantly higher than in pigs fed control diet(P <0.05). Calcium digestibility in pigs fed P-0.2 diet was significantly higher than in pigs fed control and P-O.I diets(P <0.05). Fecal-N excretion was lower in pigs fed P-O.! and P-0.2 diets than in pigs fed control(P <0.05). In the second metabolic trial(41.80 $\pm$ 2.68kg average body weight), there was no significant difference among treatments in apparent digestibility of nutrients and N-retention. In blood assay for the BUN and immune responses investigations, there was no significant difference among treatments during overall period of experiment. Therefore, this experiment suggested that probiotics supplementation could improve growth performance and nutrient digestibility of pigs.

Antimutagenic and Antitumor Effects of Adenophora triphylla Extracts (잔대 추출물들의 항돌연변이 및 항종양 효과)

  • Ham, Young-An;Choi, Hyun-Jin;Kim, Soo-Hyun;Chung, Mi-Ja;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.1
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    • pp.25-31
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    • 2009
  • This study was carried out to investigate the mutagenic, antimutagenic, cytotoxicity and antitumor effects of Adenophora triphylla (AT). AT was extracted with 70% ethanol and then further fractionated to hexane, chloroform, ethyl acetate, butanol and water. Antimutagenic, cytotoxicity and antitumor effects of AT extracts were measured by using Ames test, SRB method, and the tumor growth inhibition test. AT extracts did not show any mutagenicity in the Ames test; however, 70% ethanol extracts and its fractions had strong antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The ethyl acetate fraction of AT (200 ${\mu}g$/plate) showed approximately 66.5% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain, whereas 83.3% and 75.1% inhibitions were observed on the mutagenesis induced by MNNG and 4NQO against TA100 strain. In anticancer effects, the cytotoxicity of AT extract and its fractions against cancer cell lines including human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (Hep3B), human breast adenocarcinoma (MCF-7), human gastric carcinoma (AGS), human lung carcinoma (A549) and transformed primary human embryo kidney (293) were investigated. The treatment of 1 mg/mL AT ethyl acetate faction had the highest cytotoxicity of 79.9%, 74.9%, 66.0%, 71.0% and 74.3% against HeLa, Hep3B, MCF-7, AGS and A549 cells, respectively. In contrast, the extract and its fractions showed only $3{\sim}36%$ cytotoxicity for a normal human kidney cell line (293). In vivo anti-cancer effect of Adenophora triphylla extract was tested using Balb/c mice transplanted sarcoma-180 cells. Adenophora triphylla ethyl acetate fraction showed the highest inhibition rate of 37.2% at the 50 mg/kg concentration.

Understanding of Phytoplankton Community Dynamics Through Algae Bioassay Experiment During Winter Season of Jinhae bay, Korea (생물검정실험을 통한 동계 진해만 식물플랑크톤의 군집 변동 특성 파악)

  • Hyun, Bong-Gil;Shin, Kyoung-Soon;Kim, Dong-Sun;Kim, Young-Ok;Joo, Hae-Mi;Baek, Seung-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.16 no.1
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    • pp.27-38
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    • 2011
  • The distributions of phytoplankton assemblages and environmental factors in Jinhae Bay and their relationships were investigated to estimate the potential limiting nutrient for phytoplankton growth and community structure. In situ algal bioassay experiments were also conducted to assess the species-specific characteristics in phytoplankton responses under different nutrient conditions (control, N(+) and P(+) treatment). During the study periods, bacillariophyceae and cryptophyceae occupied more than 90% of total phytoplankton assemblages. Phytoplankton standing crops in the inner part of Masan Bay were higher than that of Jinhae Bay. The DIN:DIP ratio, pH and transparency showed the significant positive correlation with phytoplankton biomass. According to cluster and multidimensiolnal scaling (MDS) analysis based on phytoplankton community data from each station, the bay was divided into three groups. The first group included stations from the south-western part of Jinhae bay where cryptophyta species were dominated. The second group was distinguished from inner stations in Masan Bay. These stations showed low transpancy and high DIN:DIP ratio. The other cluster included the stations from the eastern part and central part of Jinhae Bay, which was characterized by the high DSi:DIP ratio and dominant of diatom species. Phosphorous (P) was limited in Masan Bay due to significantly increases in the phytoplankton abundances. Based on stoichiometric limitation and algal bio-assay in Jinhae Bay, nitrogen (N) was a major limiting factor for phytoplankton production. However, silicate (Si) was not considered as limiting factor, since Si/DIN and Si/P ratio and absolute concentration of nutrient did not create any potential stoichiometric limitation in the bay. This implies that high Si availability in winter season contributes favorably to the maintenances of diatom species.

Significance of Serum Ferritin in Multiple Trauma Patients with Acute Respiratory Distress Syndrome (다발성 외상 환자에서 발생되는 급성 호흡 곤란 증후군의 예측 인자로서 혈청 페리틴의 의의)

  • Ji, Yae-Sub;Kim, Nak-Hee;Jung, Ho-Geun;Ha, Dong-Yeup;Jung, Ki-Hoon
    • Journal of Trauma and Injury
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    • v.20 no.2
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    • pp.57-64
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    • 2007
  • Purpose: Clinically, acute respiratory distress syndrome (ARDS) occurs within 72 hours after acute exposure of risk factors. Because of its high fatality rate once ARDS progresses, early detection and management are essential to reduce the mortality rate. Accordingly, studies on early changes of ARDS were started, and serum ferritin, as well the as injury severity score (ISS), which has been addressed in previous studies, thought to be an early predictive indicator for ARDSMethods: From March 2003 to March 2005, we investigated 50 trauma patients who were admitted to the intensive care unit in Dongguk University Medical Center, Gyeongju. The patients were characterized according to age, sex, ISS, onset of ARDS, time onset of ARDS, serum ferritin level (posttraumatic $1^{st}\;&\;2^{nd}$ day), amount of transfused blood, and death. Abdominal computed topography was performed as an early diagnostic tool to evaluate the onset of ARDS according to its diagnostic criteria. The serum ferritin was measured by using a $VIDAS^{(R)}$ Ferritin (bioMeriux, Marcy-1' Etoile, France) kit with an enzyme-linked fluorescent assay method. For statistical analysis, Windows SPSS 13.0 and MedCalc were used to confirm the probability of obtaining a predictive measure from the receiver operating characteristics (ROC) curve. Results: The ISS varied from 14 to 66 (mean: 33.8) whereas the onset of ARDS could be predicted with the score above 30 (sensitivity: 90.0%, specificity: 60.0%, p<0.05). On the posttraumatic $1^{st}$ day, the serum ferritin levels were measured to be from 31 mg/dL to 1,200 mg/dL (mean: 456 mg/dL), and the onset of ARDS could be predicted when the value was over 340 mg/dL (sensitivity: 80.0%, specificity: 65.0%, p<0.05). On the posttraumatic $2^{nd}$ day, the serum ferritin levels were measured to be from 73 mg/dL to 1,200 mg/dL (mean: 404 mg/dL), and the onset of ARDS could be predicted when the value was over 627 mg/dL (sensitivity: 60.0%, specificity: 92.5%, p<0.05). The serum ferritin levels and the ISS were significantly higher on the posttraumatic $1^{st}$ and $2^{nd}$ day in the ARDS group, suggesting that they are suitable indices predicting the onset of ARDS, however relationship between the serum ferritin levels and the ISS was not statistically significant. Conclusion: In this study, we discovered increasing serum ferritin levels in multiple- trauma patients on the posttraumatic $1^{st}$ & $2^{nd}$ day and concluded that both the serum ferritin level and the ISS were good predictors of ARDS. Although they do not show statistically significant relationship to each other, they can be used as independent predictive measures for ARDS. Since ARDS causes high mortality, further studies, including the types of surgery and the methods of anesthesia on a large number of patients are essential to predict the chance of ARDS earlier and to reduce the incidence of death.

Fatty acid analysis and regulatory effects of citron (Citrus junos Sieb. ex TANAKA) seed oil on nitric oxide production, lipid accumulation, and leptin secretion (유자씨유의 지방산분석 및 Nitric Oxide 생성, 지방축적능, 렙틴분비 조절효과)

  • Kim, Tae Woo;Kim, Kyoung Kon;Kang, Yun Hwan;Kim, Dae Jung;Choe, Myeon
    • Journal of Nutrition and Health
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    • v.47 no.4
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    • pp.221-228
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    • 2014
  • Purpose: Citron seed oil (CSO) has been reported to have high antioxidant activity. However, the composition and other biologically activities of CSO have not been reported. In this study, we confirmed the fatty acid composition of CSO, which may be beneficial to vascular disease and obesity. Methods: We investigated the oil composition of CSO using gas chromatography coupled with mass spectrometry (GC-MS) analysis, and cytotoxicity was confirmed by Cell Counting Kit-8 (CCK-8) assay. Nitric oxide (NO) production in human umbilical vein endothelial cells (HUVECs) was measured using Griess reagent, and lipid accumulation and leptin secretion in 3T3-L1 cells were measured by Oil-Red O staining and commercial ELISA kit, respectively. Results: GC-MS analysis indicated that CSO contains several components, including linoleic acid, oleic acid, palmitic acid, stearic acid, linolenic acid, palmitoleic acid, and arachidic acid. In physiological activity analysis, CSO did not induce cytotoxic effects in HUVECs and 3T3-L1 cells. Further, CSO significantly induced nitric oxide and leptin secretion as well as inhibited lipid accumulation. Conclusion: CSO increased NO release, inhibited lipid accumulation, and induced leptin secretion, suggesting it may be useful for the management of vessels and weight gain. Although further studies are required to investigate the safety and mechanism of action of CSO, our results show that the composition and physiological activity of CSO are sufficient for its use as functional edible oil.

Development of salt-tolerant transgenic chrysanthemum (Dendranthema grandiflorum) lines and bio-assay with a change of cell specificity (내염성 국화 형질전환 계통 육성 및 저항성 검정과 세포특성 변화)

  • Kang, Chan-Ho;Yun, Seung-Jung;Han, Bum-So;Lee, Gong-Joon;Choi, Kyu-Hwan;Park, Jong-Suk;Shin, Yong-Kyu
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.1-8
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    • 2011
  • Recently the increasing of vinyl and green houses and development of reclaimed land including Saemangeum induced the need for breeding salt-tolerant crops which can survive and grow in high salinity soil. So we try to develop salt-tolerant transgenic chrysanthemum (Dendranthema grandiflorum.) lines by using anti-porter gene TANHX and HVNHX. Through marker selection and plant regeneration step, we could get 284 putative transgenic chrysanthemum lines. On selected putative transgenic plants, 40 candidates were used for genetic analysis and 30 lines could be made up of target size band on PCR, so about 75% of marker selected lines were decided as real transgenic lines. Selected 284 transgenic lines were also used for salt-tolerance test as a range of NaCl 0.2 ~ 1.2% (300 mM). As a result of salt-tolerance test, 15 selected transgenic lines could live and grow on the continuous supply of 0.8% (200 mM) NaCl solution and another 7 lines were could survive under 1.2% (300 mM) NaCl solution. This salt-tolerant transgenic lines under salt stress also lead a cell alternation especially a guard cell. A stressed guard cell be swelled and grow larger in proportion to NaCl concentration. TTC test for cell viability on transgenic chrysanthemum lines pointed out that more strong salt-tolerant lines can be live more than another under same salt stress. The numerical value of strong salt-tolerant 7 transgenic lines were 0.206 ~ 0.331 under 1.2% NaCl stress, and then it's value is more larger than middle salinity lines' 0.114 ~ 0.193 and non-transgenic's 0.046. And the proline contents as indicated stress compound also pointed out that HVNHX introduced salt-tolerant transgenic lines were less stressed than other under same salt stress. The contents of strong salt-tolerant transgenic lines were 2.255 ~ 2.638 mg/kg and it is much higher than that of middle salinity lines' 1.496 ~ 2.125.

The Influence of Nutrients Addition on Phytoplankton Communities Between Spring and Summer Season in Gwangyang Bay, Korea (광양만에서 춘계와 하계 영양염류 첨가가 식물플랑크톤군집의 성장에 미치는 영향)

  • Bae, Si Woo;Kim, Dongseon;choi, Hyun-Woo;Kim, Young Ok;Moon, Chang Ho;Baek, Seung Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.19 no.1
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    • pp.53-65
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    • 2014
  • In order to estimate the effect of nutrients addition for phytoplankton growth and community compositons in spring and summer season, we investigated the abiotic and biotic factors of surface and bottom waters at 20 stations of inner and offshore areas in Gwangyang Bay, Korea. Nutrient additional experiments were also conducted to identify any additional nutrient effects on phytoplankton assemblage using the surface water for the assay. Bacillariophyceae occupied more than 90% of total phytoplankton assembleges. Of these, diatom Eucampia zodiacus and Skeletonema costatum-like species was mainly dominated in spring and summer, respectively. Here, we can offer the season why the two diatom population densities were maintained at high levels in both seasons. First, light transparency of spring season in the euphotic zone was greatly improved in the bay. This improvement is one of important factor as tigger of increase in E. zodiacus population. Second, low salinity and high nutrient sources supplied by Seomjin River discharge are a main cue for strong bottom-up effects on S. costatum-like species during the summer rainy season. Based on the algal bio-assays, although maximum growth rate of phytoplankton communities at inner bay (St.8) were similar to those of outer bay (St.20), half-saturation constant ($K_s$) for phosphate at outer bay was slightly lower than those of inner bay. This implied that adapted cells in low nutrient condition of outer bay may have enough grown even the low phosphate and they also have a competitive advantage against other algal species under low nutrient condition. In particular, efficiency of N (+) addition in summer season was higher compared to control and P added experiments. In the bay, silicon was not a major limiting factor for phytoplankton growth, whereas nitrogen (N) was considered as a limiting factor during spring and summer. Therefore, a sufficient silicate supply form water mixing Si recycled from diatom decomposition and river water is favorable form maintaining diatom ecosystems in Gwangyang Bay.

Short-term changes of phytoplankton communities after nutrient addition and establishment of stable mass culture condition to prepare the type approval test of USCG Phase-II in mesocosm enclosure (메소코즘에서 USCG phase-II 형식승인 대비 영양염 첨가에 따른 식물플랑크톤 대량 배양조건 확립 및 군집구조의 단주기변화)

  • Baek, Seung Ho;Lee, Min Ji;Shin, Kyoungsoon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.4
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    • pp.34-42
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    • 2016
  • In order to prepare for the type approval test for the United States Coast Guard (USCG) Phase-II of Ballast Water Treatment System (BWTS), a phytoplankton mass culture was conducted in a mesocosm enclosure. We evaluated the response of the phytoplankton community after nutrient addition (+N, +P, and +NP) and investigated the development of the species with increasing culture time. After nutrient dosing, the phytoplankton population significantly (p < 0.05) increased from day 1 to day 3, depending on the nutrient treatments In particular, the specific growth rate of the phytoplankton community in the case of +NP treatment and + N treatment were estimated to be $2.47d^{-1}$ and $1.98d^{-1}$, respectively. The phytoplankton population density in the case of + NP treatment was approximately 50 times higher than that of the control group, suggesting that these treatments could be useful for mass culturing phytoplankton (> 75% of natural community) for the approval regulation of USCG Phase-II. In the phytoplankton community of the mesocosm, Pseudo-nitzchia spp. dominated in the logarithmic growth phase. The cell density decreased significantly (p < 0.05) with increasing time, coinciding with the nutrient limitation. At that time, the dominance of Pseudo-nitzchia spp. shifted to that of Cylindrotheca closterium. Therefore, the optimum nutrient concentration ($N:30{\mu}M$, $P:3{\mu}M$) and reasonable harvesting time (after 3 days in summer) found in this study for the mass culturing of phytoplankton may be helpful to meet the USCG Phase-II biological criteria to be used in BWTS.

Antioxidant capacity and Raw 264.7 macrophage anti-inflammatory effect of the Tenebrio Molitor (갈색거저리(Tenebrio Molitor)의 항산화능과 Raw 264.7 대식세포의 항염증 효과)

  • Yu, Jae-Myo;Jang, Jae-Yoon;Kim, Hyeon-Jeong;Cho, Yong-Hun;Kim, Dong-in;Kwon, O-jun;Cho, Yeong-Je;An, Bong-Jeun
    • Food Science and Preservation
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    • v.23 no.6
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    • pp.890-898
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    • 2016
  • The purpose of this paper is to investigate potential anti-inflammatory and anti-oxidant effects of Tenebrio molitor. Macrophage cell response by outside stimulation leads expression of pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin-6 (IL-6), $interleukin-1{\beta}$ ($IL-1{\beta}$), and trigger expression of genes which are affected by inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), resulting in formation of inflammatory factors like nitric oxide (NO) and Prostaglandin $E_2$ (PGE2). Cell viability was determined by MTT assay. In order to investigate anti-inflammatory agents, the inhibitory effects on the production of lipopolysaccharide (LPS)-induced NO in RAW 264.7 cells were examined. T. Molitor significantly decreased the production of NO in a dose-dependent manner, and also reduced the expression of iNOS, a COX-2 protein. As a result, the levels of protein such as $PGE_2$, iNOS, COX-2 and MARKs were significantly reduced compared to non-treated group in T. Molitor water extract (TDW) treated group. Also, antioxidant effect of T. Molitor were investigated using DPPH, ABTS+ and superoxide anion radical scavenging activity tests in cell-free system. Antioxidant activity of T. molitor was found low in the DPPH radical scavenging test while high in the ABTS+ and superoxide anion radical scavenging activity tests. These results show that TDW could be an effective anti-pro-inflammatory and anti-oxidant agent.