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Effects of Culture Media and Oxygen Concentration on In Vitro Development of Porcine IVM/IVE Embryos (배양액 및 산소농도가 돼지 체외수정란의 발달에 미치는 영향)

  • Choe, C.Y.;Choe, S.R.;Choi, S.H.;Kim, H.J.;Han, M.H.;Kang, D.W.;Shin, Y.W.;Han, J.H.;Son, D.S.
    • Journal of Embryo Transfer
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    • v.22 no.3
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    • pp.155-160
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    • 2007
  • During in vitro culture of mammalian oocytes and embryos, the cells are exposed to the risks that cause cell injury or death. Numerous studies have been reported that the cell injury may be induced by the action of free radicals generated by auto-oxidation. This study was undertaken to investigate the optimal culture condition system for in vitro culture of porcine embryos. We first evaluated the effect of culture media on the porcine embryo development. NCSU-23 and PZM-5, culture medium tested, were failed to produce significant difference on the rate of blastocyst formation. In NCSU-23, the developmental rate was slightly higher than that in PZM-5. During in vitro maturation (IVM), fertilizaton (IVF), and culture (IVC) under 5 or 20% oxygen ($O_2$), the rates of cleavage and development were insignificantly different from each other under our culture condition (20% $O_2$, in NCSU-23), the mean cell number per blastocyst was $40{\pm}10$. These results showed that medium and $O_2$ concentration had no significant effect on the development of porcine embryos.

Antioxidant, antibacterial, antifungal, and anti-inflammatory effects of 15 tree essential oils (수목 방향유 15종의 산화방지, 항세균, 항진균 및 항염증 효과)

  • Jo, Se Jin;Park, Mi-Jin;Guo, Rui Hong;Park, Jung Up;Yang, Ji Yoon;Kim, Jae-Woo;Lee, Sung-Suk;Kim, Young Ran
    • Korean Journal of Food Science and Technology
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    • v.50 no.5
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    • pp.535-542
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    • 2018
  • The current study investigated the beneficial effects of 15 essential oils isolated from tree branches, leaves, and flowers. Among these oils, clove bud and Illicium anisatum oils showed the most potent anti-oxidant effects on 1,1-diphenyl-2-picrylhydrazyl and 2,2'azinbis-(3-ethyl-benzothiazoline-6-sulfonic acid) radical scavenging activities. Next, we evaluated the antibacterial effects of 15 essential oils on Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7, Salmonella typhimurium, and Streptococcus mutans. Clove bud significantly decreased growth of 5 bacterial strains. In addition, clove bud, Magnolia kobus, Picea abies and Chamaecyparis obtuse significantly reduced growth of the fungi, Aspergillus fumigatus, Aspergillus ochraceus, Candida albicans and Trichophyton rubrum. Additionally, clove bud also remarkably reduced the expression of cyclooxygenase-2 and inducible NO synthase in lipopolysaccharide-activated RAW264.7 cells. These results indicate that essential oils isolated from trees, which exhibit antioxidant, antibacterial, antifungal and anti-inflammatory properties, may be potentially useful in the development of cosmetic ingredients.

Physico-chemical characteristics and ${\beta}-galactosidase$ activity of Lactobacillus plantarum from kimchi (김치에서 분리한 유산균 Lactobacillus plantarum의 이화학적 특성 및 ${\beta}-galactosidase$ 활성)

  • Kang, Mi-Seon;Rhee, Young-Hwan
    • Applied Biological Chemistry
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    • v.39 no.1
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    • pp.54-59
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    • 1996
  • Three strains of inhibitory lactic acid bacteria (No. 49, No. 61, No. 75) against Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escheirchia coli(ATCC33694) and Bacillus subtilis(ATCC6633) were isolated from kimchi, and then, identified to be Lactobacillus plantarum after examinations of their biological and physiological characteristics. To investigate a possible application of these three lactobacilli in milk fermentation industry, we made yogurts and then evaluated their ${\beta}-galactosidase$ activities at various; incubation time(from 24 hrs to 72 hrs). The result of experiment was that ${\beta}-galactosidase$ activities were reached maximum at 48 hrs and that reduced gradually with the lapse of time. And the ${\beta}-galactosidase$ activity of lactobacilli, and their viable cell counts at $37^{\circ}C$ for 2 hrs under various pH conditions were investigated. ${\beta}-galactosidase$ activities of 3 strains were reduced 50% at pH 3.5, but there were no remaining activities at pH 2.5, and pH 1.5, respectively. The frequency of the survival cell of lactobacilli in yogurt were $0.12{\sim}0.75%$ at pH 2.5, $$6.3{\times}10^{-5}{\sim}2.7{\times}10^{-3}% at pH 1.5, respectively, but there was no significant difference at pH 3.5. The values of original pH, titratable acidity as lactic acid, viscosity, and viable cells of yogurts were $4.08{\sim}4.30,\;1.05{\sim}1.25%,\;1,818{\sim}2,124\;cps\;and\;7.3{\times}10^8{\sim}3.0{\times}10^9\;cfu/m{\ell}$, respectively. To estimate buffer capacity of yogurt, the volume of 1.0 N HCl to 2 unit below original pH of yogurt($100\;m{\ell}$) was $11.98{\sim}13.02\;m{\ell}$ and the volume of 1.0N NaOH to 4 unit above original pH of yogurt($100\;m{\ell}$) was $10.82{\sim}12.86\;m{\ell}.

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Development of Yoghurt with Sanmeoru (Vitis amurensis Ruprecht) Wine as an Additive (산머루 와인을 첨가한 요구르트의 품질 특성)

  • Kim, Jae-Kyeoung;Lee, Jai-Sung;Jeong, Yu-Tae;Bae, In-Hyu
    • Journal of Dairy Science and Biotechnology
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    • v.30 no.1
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    • pp.23-30
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    • 2012
  • This study was carried out to investigate the quality characteristics of yoghurt with varying contents of Sanmeoru (Vitis amurensis Ruprecht) wine (SW). Yoghurts were fermented with 2 kinds of lactic acid bacteria (Streptococcus thermophilus and Lactobacillus bulgaricus; Yo-$MIX^{TM5}505$, Danisco, Denmark). The changes in quality characteristics were investigated during fermentation and storage, and anthocyanin composition was analyzed after fermentation. The pH decreased in all treatments, while titratable acidity and viscosity gradually increased during fermentation. The viscosity of the yoghurt with 5.0% cultured SW added was higher than in other groups. The viable counts of lactic acid bacteria gradually increased during fermentation. The sample of the 5.0% cultured SW-yoghurt was higher than that of the other groups. The pH slowly decreased during the storage period; this was lower in the control yoghurt than in the yoghurts containing SW, and the changes of titratable acidity in the samples containing 5.0% and 30.0% cultured SW were lower than that in the yoghurt containing 1.0% cultured SW and in the control. The change in viscosity and number of viable cells during storage was higher in the yoghurts containing SW than in the control group. The levels of anthocyanin increased with the SW content in the yoghurt. Sensory scores for taste of the yoghurt with 1.0% added SW was significantly higher than for the other groups. Yoghurt with added SW kept at $4^{\circ}C$ for 12 days retained its quality characteristics fairly well.

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Anti-inflammatory effects of Ishige sinicola ethanol extract in LPS-induced RAW 264.7 cell and mouse model (LPS로 유도된 RAW 264.7 Cell과 마우스 모델에 대한 넓패(Ishige sinicola) 에탄올 추출물의 항염증 효과)

  • Kim, Ji-Hye;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Cho, Kwang-Su;Kim, Go-Eun;XU, Xiaotong;Lee, Da-Hye;Park, Ga-Ryeong;Ahn, Dong-Hyun
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1149-1157
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    • 2017
  • Inflammation is the first response of the immune system to infection or irritation in our body. The use of medicinal plants has been widely applied as an alternative source for drug development. One of marine natural resources, the anti-inflammatory effect of Ishige sinicola ethanol extract (ISEE), was evaluated by using LPS-induced RAW 264.7 cell and mice model. As a result, the production of nitric oxide (NO) and pro-inflammatory cytokines (IL-6, IL-$1{\beta}$, TNF-${\alpha}$) were inhibited with increasing concentration of ISEE without any cytotoxicity. Furthermore, ISEE suppressed the expression of not only inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-${\kappa}B$) p65, and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK) in a dose-dependent manner. In mice ear edema test, the formation of edema was reduced at the highest dosage of ISEE and the reduction of the number of infiltrated mast cells was observed in histological analysis. These results indicate that ISEE has a potent anti-inflammatory activity and can be used as a pharmaceutical material for many kinds of inflammatory disease.

The Effects of Mycobacterium Tuberculosis on Alveolar Macrophages -The Alterations of Superoxide Production in both Human and Rat Alveolar Macrophages Exposed to Mycobacterium Tuberculosis H37Ra Strain- (결핵균이 폐포대식세포의 기능에 미치는 영향에 관한 연구 -H37Ra 결핵균종에 의한 사람 몇 백서 폐포대식세포의 Superoxide 생성의 변화-)

  • Kim, Keon-Youl;Lee, Kye-Young;Hyun, In-Kyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.6
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    • pp.526-535
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    • 1992
  • Background: The oxygen radicals released by alveolar macrophages contribute to killing of microorganisms including M. tuberculosis. Macrophages are "primrd" for enhanced oxygen radical release by macrophage activator like IFN-$\gamma$ and LPS, which do not themselves cause release of oxygen radicals. Actural production of oxygen radicals is "triggered" by phagocytosis or by exposure to chemical stimuli like PMA or FMLP. There has been debates about the priming effect of alveolar macro phages because they are exposed to usual environmental particles unlike blood monocytes. Therefore we examined priming effect of IFN-$\gamma$ in human alveolar macrophages comparing with that in blood monocytes and rat alveolar macrophages. And we observed the alterations of superoxide production in both human and rat alveolar macrophages after exposure to M. tuberculosis H37Ra bacilli itself and its lysate. Methods: Bronchoalveolar lavage fluid was processed to isolate alveolar macrophages by adherence and the adherent cells were removed by cold shock method. After exposure to M. tuberculosis H37Ra strain, alveolar macrophages were incubated for 24 hours with IFN-$\gamma$. The amount of superoxide production stimulated with PMA was measured by ferricytochrome C reduction method. Results: 1) The priming effect in human alveolar macrophages was not observed even with high concentration of IFN-$\gamma$ while it was observed in blood monocytes and rat alveolar macrophages. 2) Both human and rat alveolar macrophages exposed to avirulent H37Ra strain showed triggering of superoxide release and similar results were shown with the exposure to H37Ra lysate. Conclusion: The priming effect in human alveolar macrophages is not observed because of its usual exposure to environmental particles and avirulent H37Ra strain does not inhibit the activation of alveolar macrophages.

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Soil Moisture Influence on Growth of Cover Vegetations and Water Economy (토양수분(土壤水分)이 피복용식물(被覆用植物)의 생장(生長) 및 수분경제(水分經濟)에 미치는 영향(影響))

  • Lee, Soo Wook
    • Journal of Korean Society of Forest Science
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    • v.33 no.1
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    • pp.1-32
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    • 1977
  • This study has been made to find out more effective way of vegetation establishment on severely denuded forest land from the viewpoint of soil moisture regimes. Various environmental factors were measured to estimate soil moisture conditions of different sites. Soil moisture influence on growth of over vegetations, water requirement and drought resistance were analyzed. The efficiency of water use was also reckoned at different fertility levels and different soil moisture conditions. This research is composed of field experiment and green house experiment. Field experiment includes height growth, survival and coverage analysis of cover vegetations (Robinia pseudoacacia L., Lespedeza bicolor Turcz, Arundinella hirta Tanaka var. ciliare Koidzumi.) with 4 fertility level treatments on 3 slopes (Steep: $37^{\circ}$, Moderate: $25^{\circ}$, Gentle; $17^{\circ}$) during dry season (1 April-30 June) and wet season (1 July-10 September). At the same time temperature, relative humidity and precipitation were measured to understand the environmental changes. Soil moisture conditions were measured with soil moisture meter with 24 soil cells. Green house experiment comprised height, fresh weight and dry weight measurements of cover vegetations with 4 fertility levels and 3 moisture conditions for 70 days. The results extracted from experiments are as follews. 1. Cover vegtations have 3 patterns of water requirement at the early stage of growth. a) Robinia type has high water requirement and weaker drought resistance. b) Lespedeza type has low water requirement and stronger drought resistance. c) Arundinella type has moderate water requirement and weaker drought resistance. 2. The vegetations have different optimum fertility levels in different soil moisture supply condition. a) Robinia needs a low fertility level in dry condition and a high level in wet condition. b) Lespedeza needs only low fertility level in all conditions. c) Arundinella needs a low fertility level in dry condition and a high level in wet condition. 3. The efficiency of water use (Water/1g dry weight) by fertility levels is different from one another. Robinia and Arundinella have a good efficiency at low fertility level in dry condition and at high fertility level in wet condition. Lespedeza has a good efficiency at low fertility level in all conditions. 4. $P_2O_5$ requirement of Robinia and Lespedeza is high, but that of Arundinella is low. Soil moisture condition has a great influence on $P_2O_5$ absorption in dendued forest land. Once Vegetations are established on suitable sites with optimum fertitity level according to different moisture condition, even the small amount of soil water in denuded land can he used with high efficiency and the effect of fertility treatment can be maximized.

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The Effect of Heat Shock Response on the Tumor Necrosis Factor-$\alpha$-induced Acute Lung Injury in Rats (Tumor Necrosis Factor-$\alpha$로 유도되는 백서의 급성 폐손상에 열충격반응이 미치는 효과)

  • Koh, Youn-Suck;Lim, Chae-Man;Kim, Mi-Jung;Cho, Won-Kyung;Jeoung, Byung-O;Song, Kyu-Young;Shim, Tae-Sun;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1343-1352
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    • 1997
  • Background : Heat-treated cells are known to be protected from lysis by TNF, which is considered to play a central role in the pathogenesis of sepsis-induced acute lung injury. The objective of the study was to investigate the effect of heat shock response by heat-pretreatment on the acute lung injury of the rats induced by intratracheally administered TNF-$\alpha$, Methods : We intratracheally instilled either saline or TNF (R&D, 500ng) with and without heat pretreatment in Sprague-Dawley rats weighing 250~350 g. The heated rats were raised their rectal temperature to $41^{\circ}C$ and was maintained thereafter for 13 minutes at 18 h before intratracheal administration of saline or TNF. After 5 h of intratracheal treatment, lung leak, lung myeloperoxidase activity (MPO) and heat shock proteins were measured in rats. Lung leak index was defined as counts per minute of $I^{25}$ in the right lung divided by counts per minutes of $I^{25}$ in 1.0 ml of blood. All data are expressed as means ${\pm}$SE. Results : There is no difference in acute lung leak index ($0.099{\pm}0.024$ vs $0.123{\pm}0.005$) among the rats given saline intratracheally with and without heat pretreatment, but MPO activity showed a decreased tendency in heat-pretreated rats ($4.58{\pm}0.79\;U/g$) compared with heat-unpretreated rats ($7.32{\pm}0.97\;U/g$) (P=0.064). Rats administered TNF intratracheally with heat-pretreatment had decreased lung leak index ($0.137{\pm}0.012$) and lung MPO activity ($5.51{\pm}1.04\;U/g$) compared with those of heat-unpretreated and TNF-administered rats ($0.186{\pm}0.016$, $14.34{\pm}1.22\;U/g$) (P<0.05 in each). There were no significant difference of lung leak index and MPO activity between TNF-treated rats with heat-pretreatment and saline-treated rats with and without heat-pretreatment. Conclusion : The heat shock response attenuated neutrophil recruitment and acute lung leak induced by intratracheal instillation of TNF-in rats.

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Evaluation of the Radioimmunotherapy Using I-131 labeled Vascular Endothelial Growth Factor Receptor2 Antibody in Melanoma Xenograft Murine Model (흑색종에서의 I-131표지 혈관내피세포성장인자 수용체2항체를 이용한 방사면역치료 평가)

  • Kim, Eun-Mi;Jeong, Hwan-Jeong;Park, Eun-Hye;Cheong, Su-Jin;Lee, Chang-Moon;Jang, Kyu-Yun;Kim, Dong-Wook;Lim, Seok-Tae;Sohn, Myung-Hee
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.4
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    • pp.307-313
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    • 2008
  • Purpose: Vascular endothelial growth factor (VEGF) and its receptor, fetal liver kinase 1 (Flk-1), play an important role in vascular permeability and tumor angiogenesis. The aim of this study is to evaluate the therapeutic efficacy of $^{131}I$ labeled anti-Flk-1 monoclonal antibody (DC101) on the growth of melanoma tumor, which is known to be very aggressive in vivo. Materials and Methods: Balb/c nude mice were injected subcutaneously with melanoma cells in the right flank. Tumors were allowed to grow up to $200-250\;mm^3$ in volume. Gamma camera imaging and biodistribution studies were performed to identify an uptake of $^{131}I$-DC101 in various organs. Mice with tumor were randomly divided into five groups (10 mice per group) and injected intravenously; control PBS (group 1), $^{131}I$-DC101 $50\;{\mu}g/mouse$ (group 2), non-labeled DC101 $50\;{\mu}g/mouse$ (group 3), $^{131}I$-DC101 $30\;{\mu}g/mouse$ (group 4) and $15\;{\mu}g/mouse$ (group 5) every 3 or 4 days for 20 days. Tumor volume was measured with caliper twice a week. Results: In gamma camera images, the uptake of $^{131}I$-DC101 into tumor and thyroid was increased with time. Biodistribution results showed that the radioactivity of blood and other major organ was gradually decreased with time whereas tumor uptake was increased up to 48 hr and then decreased. After 4th injection of $^{131}I$-DC101, tumor volume of group 2 and 4 was significantly smaller than that group 1. After 5th injection, the tumor volume of group 5 also significantly reduced. Conclusion: These results indicated that delivery of $^{131}I$ to tumor using FlK-1 antibody, DC101, effectively blocks tumor growth in aggressive melanoma xenograft model.

Antimicrobial, Antioxidant and Cellular Protective Effects against Oxidative Stress of Anemarrhena asphodeloides Bunge Extract and Fraction (지모 뿌리 추출물과 분획물의 항균활성과 항산화 활성 및 세포보호 연구)

  • Lee, Yun Ju;Song, Ba Reum;Lee, Sang Lae;Shin, Hyuk Soo;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.360-371
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    • 2018
  • Extracts and fractions of Anemarrhena asphodeloides Bunge were prepared and their physiological activities and components were analyzed. Antimicrobial activities of the ethyl acetate and aglycone fractions were $78{\mu}g/ml$ and $31{\mu}g/ml$, respectively, for Staphylococcus aureus and $156{\mu}g/ml$ and $125{\mu}g/ml$, respectively, for Pseudomonas aeruginosa. 1,1-Diphenyl-2-picrylhydrazyl free radical scavenging activities ($FSC_{50}$) of 50% ethanol extract, ethyl acetate fraction, and aglycone fraction of A. asphodeloides extracts were $146.2{\mu}g/ml$, $23.19{\mu}g/ml$, and $71.06{\mu}g/ml$, respectively. The total antioxidant capacity ($OSC_{50}$) in an $Fe^{3+}$-EDTA/hydrogen peroxide ($H_2O_2$) system were $17.5{\mu}g/ml$, $1.5{\mu}g/ml$, and $1.4{\mu}g/ml$, respectively. The cytoprotective effect (${\tau}_{50}$) in $^1O_2$-induced erythrocyte hemolysis was 181 min with $4{\mu}g/ml$ of the aglycone fraction. The ${\tau}_{50}$ of the aglycone fraction was approximately 4-times higher than that of (+)-${\alpha}$-tocopherol (${\tau}_{50}$, 41 min). Analysis of $H_2O_2$-induced damage of HaCaT cells revealed that the maximum cell viabilities for the 50% ethanol extract, ethyl acetate fraction, and aglycone fraction were 86.23%, 86.59%, and 89.70%, respectively. The aglycone fraction increased cell viability up to 11.53% at $1{\mu}g/ml$ compared to the positive control treated with $H_2O_2$. Analysis of ultraviolet B radiation-induced HaCaT cell damage revealed up to 41.77% decreased intracellular reactive oxygen species in the $2{\mu}g/ml$ aglycone fraction compared with the positive control treated with ultraviolet B radiation. The findings suggest that the extracts and fractions of A. asphodeloides Bunge have potential applications in the field of cosmetics as natural preservatives and antioxidants.