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Echinacoside Induces UCP1- and ATP-Dependent Thermogenesis in Beige Adipocytes via the Activation of Dopaminergic Receptors

  • Kiros Haddish;Jong Won Yun
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1268-1280
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    • 2023
  • Echinacoside (ECH) is a naturally occurring phenylethanoid glycoside, isolated from Echinacea angustifolia, and this study aimed to analyze its effect on thermogenesis and its interaction with dopaminergic receptors 1 and 5 (DRD1 and DRD5) in 3T3-L1 white adipocytes and mice models. We employed RT-PCR, immunoblot, immunofluorescence, a staining method, and an assay kit to determine its impact. ECH showed a substantial increase in browning signals in vitro and a decrease in adipogenic signals in vivo. Additionally, analysis of the iWAT showed that the key genes involved in beiging, mitochondrial biogenesis, and ATP-dependent thermogenesis were upregulated while adipogenesis and lipogenesis genes were downregulated. OXPHOS complexes, Ca2+ signaling proteins as well as intracellular Ca2+ levelswere also upregulated in 3T3-L1 adipocytes following ECH treatment. This was collectively explained by mechanistic studies which showed that ECH mediated the beiging process via the DRD1/5-cAMP-PKA and subsequent downstream molecules, whereas it co-mediated the α1-AR-signaling thermogenesis via the DRD1/5/SERCA2b/RyR2/CKmt pathway in 3T3-L1 adipocytes. Animal experiments revealed that there was a 12.28% reduction in body weight gain after the ECH treatment for six weeks. The effects of ECH treatment on adipose tissue can offer more insights into the treatment of obesity and metabolic syndrome.

Survey for Approximate Composition and Mineral Content of Medicinal Herbs (약초중의 일반성분 및 무기질 함량조사)

  • Hwang, Jin-Bong;Yang, Mi-Ok;Shin, Hyung-Kyung
    • Korean Journal of Food Science and Technology
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    • v.29 no.4
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    • pp.671-679
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    • 1997
  • Approximate composition and mineral content of eighty seven medicinal herbs marketed in Korea was analyzed on the basis of their dried weight. From the results anlyzed, it was known that Acanthopanax sessiliflorum Seem. (22.4%), Portulaca oleracea L. (20.6%), Torilis japonica DC. (15.0%) in crude ash, Ziziyphus spinosa Hu. (39.9%), Alisma orientale Juzep. (30.6%), Lepidium latifolium L. (28.8%), Cassia tora L. (27.5%) in crude protein, Ziziyphus spinosa Hu. (27.9%), Xantnium strumarium L. (24.0%), Perilla frutescens var. acuta Kude (22.8%), Benincasa hispida Cong. (22.4%) in crude lipid, Vitex rotundifolia L. (59.5%), Ganoderma lucidum Karst (53.9%), Pinus densifiora Sieb. et Zucc. (46.4%), Sambucus williamsii Hanse (41.2%) in crude fiber, and Poria cocos Wolf (91.2%), Gastrodiae elata Blume (87.9%), Gentiana scabra Bunge (86.3%), Rehnannia glutinosa Libosch (86.0%) in nitrogen free extract exhibited higher content than other kinds. Phosphrous contents were 1.5, 1.1 and 1.0% in order of Taxillus chinensis Danser, Malva verticillata L., Nelumbo nucifera Gaeten, and magnesium contents were 0.7, 0.5 and 0.5% in the order of Saururus chinesis Baill., Salivia milltriorrhiza Bunge and Pueraria thunberglans Benth. Calcium contents of Cimicifuge heradeifolia Komarov, Acanthopanax sessiliflorum Seem. and Rhus vericiflura Stokes were 8.5, 7.1 and 3.0%, and potassium contents of Tricholoma matsutake Sing., Houttuynia cordata Thunb., Achyranthes japonica Nakai were 5.6, 5,4 and 4.7%, respectively. Sodium contents were 2.1, 0.6 and 0.3% in the order of Boshinakia rossica Fedtsch., Eucommia ulmoides Oliv. and Prunus mume Seib., and iron contents were 0.3, 0.2 and 0.2% in the order of Cirsium japonicum var, ussuriense Kitamuraa, Gentiana scabra Bunge and Phlomis umbrosa Turcz.

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Investigation of radical scavenging and anti-inflammatory activity of the extract and fractions of Termitomyces albuminosus (계종버섯 추출물과 분획물의 라디칼 소거 활성과 항염증 활성 분석)

  • Kang, Min-Jae;Kim, Young-Jae;Hwang, Hyeon-Jeong;Kim, Moon-Ho;Sin, Jong-Moo;Shin, Pyung-Gyun;Kim, Gun-Do;Kim, Keun Ki
    • Journal of Mushroom
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    • v.18 no.1
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    • pp.45-52
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    • 2020
  • Termitomyces albuminosus has been recognized to have the best mushrooms in China, in terms of taste and aroma. The efficacy of these mushrooms has been recorded in the botanical list. However, research on the development of their artificial culture methods is necessary. In this study, we prepared an organic solvent extract and a hot water extract to understand the development of compounds and functional foods with antioxidant and anti-inflammatory activities. The IC50 value of DPPH radical scavenging activity of the hot water extract (TA4) was 1.5 mg/mL and the IC50 value of the MeOH fraction (TA2) was 1.93 mg/mL. The anti-inflammatory activity was investigated by the inhibition of NO production. EtOAc fraction (TA1) is a crude extract, but 79% of NO production was inhibited at 100 ㎍/mL. NO was not produced at 200 ㎍/mL. TA1-5-6, from TA1 inhibited NO production by 15% as compared to the positive control at 15 ㎍/mL, and completely inhibited NO production at 30 ㎍/mL. No cytotoxicity was observed at 50 ㎍/mL. TA2-1-5 from the MeOH fraction (TA2) inhibited more than 75% of NO production at 30 ㎍/mL; cytotoxicity was very low even at 50 ㎍/mL. In conclusion, by selective solvent selection, it was possible to manufacture an extract with no cytotoxicity and excellent biological activities. Furthermore, the extracts showed potential for developing various functional foods and drugs.

Batch and Fed-batch Fermentation for the Lovastatin Production by Cerulenin-resistant Aspergillus terreus Mutant (Cerulenin 저항성 Aspergillus terreus 변이주로부터 lovastatin 생산을 위한 회분식과 유가식 배양)

  • 문미경;전계택;정용섭
    • KSBB Journal
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    • v.16 no.1
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    • pp.87-94
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    • 2001
  • The biosynthesis of Lovastatin, a cholesterol lowering agent formed by the filamentous fungus, cerulenin-resistant Aspergillus terreus mutant was studied in shake flasks and bioreactors. The lovastatin production could be improved by fed-batch under the limited condition of carbon source. The relationship between the fungal morphology and the lovastatin production was also examined during the fed-batch cultures. The fed-batch studies in shake flasks were carried out to find the optimum glucose feeding method, and the pulsed feeding of glucose from 3 days onward at 24 hours intervals was found to be optimal to increase the lovastatin production and reduce the average pellet size. When the pH was controlled at around 5.8 during the whole fermentation period, the lovastatin concentration reached 384 mg/L, which is much higher than the values obtained pH-uncontrolled and pH 7.4. The optimal glucose feeding strategies was found that 30 g/L of glucose was added initially in batch mode, and then fed-batch was conducted by continuous addition of glucose solution(180 g/L) from 72 to 240 hr at a rate of 1.2 mL/hr at $28^{\circ}C$, pH 5.8, 400 rpm, and 1.0 vvm. The lovastatin concentration of 547 mg/L was obtained in 168 hr. It was about 1.5 times higher than the value of the batch fermentation.

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Micropropagation through Stem, Node-bud Shoot Tip and Bulblet Scale Culture in Fritillaria thunbergii Miq. (패모의 줄기, 마디, 정단 및 자구인편 배양에 의한 기내 증식)

  • Peak, Kee-Yoeup;Yu, Kwang-Jin;Seong, Nak-Sul;Choi, In-Sick;Cho, Jin-Tae
    • Korean Journal of Medicinal Crop Science
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    • v.2 no.2
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    • pp.154-161
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    • 1994
  • This experiment was carried out to establish micropropagation system in Fritillaria thunbergii Miq. Through the culture of bulblet scales, stems, node-buds and shoot tips with special reference to the effect of physiological age of explant and plant growth regulators on bulblet formation. Number of formed bulblets was significantly increased in node-bud or stem tissue compared to scals segments and on the medium supplemented with kinetin than BA containing medium. Optimum levels of kinetin for bulblet formation from node-bud taken from above 3 cm shoot length and stem segments excised from below 3 cm shoot length were 5.0 mg /L and $1.0{\sim}3.0\;mg$ /L kinetin, respectively. Interesting phenomenon was observed, the direct formation of bulblets from the axilliary bud of cultured explants. Bulblet forming capacity in stem tissue was depended on stem age, young stem had high regeneration ability compared to old stem taken from above 10 cm shoot length. 1.0 mg /L kinetin was optimum concentration for the formation of bulblets from old stem segments. Stem tissue taken from underground growing plant was promoted coampare to shoot tips or bulb scale segments. Optimum concentration of sucrose was $5{\sim}7%$. Summariged above results revealed that effective explant for micropropagation was stem and /or node-bud tissue excised from less than 3 cm plant height compared to those of bulb scale segments which showed high contamination after culture. Maximum multiplication rate of young stem and /or node-bud segment was about 20 times. Kinetin requirement for stimulation of bulblet formation from cultured explant depended on source of explants but favorable levels of kinetin for organogenesis ranged from 1.0 mg /L to 5.0 mg /L.

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Factors Influencing Protoplast Isolation and Culture in Three Capsicum Species (세 가지 고추속 식물의 원형질체 분리 및 배양에 미치는 요인)

  • 임학태;염옥희;전익조;조미애;양승균
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.141-146
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    • 2001
  • Protoplasts were isolated from cotyledons, hypocotyls, and mesophyll tissues of three species of Capsicum species (C. anuumm, C. bacatuum, and C. chacoense). Combination of Cellulysin (1%) and Macero-zyme (0.25%) in 0.65 M sorbitol was found to be the most effective for the digestion of cell wall, regardless of the Capsicum species. Antioxidant MES (2-[N-Morpholino]ethanesulfonic acid) in the enzyme solution helped protoplasts overcome browning. After 5 days of initial culture, Cell division occurred in modified K8p medium containing 1~5 mg/L zeatin, 0.5 mg/L IAA, 0.1~0.5 mg/L TDZ, and 1 mg/L 2,4-D under continuous dark condition at $25^{\circ}C$. Semi-solid agarose culture method was more effective than liquid culture, and it also protected the cells from browning caused by polyphenolic compound released during protoplast culture. A total of 4000 calli were obtained from protoplast culture of different capsicum species. All of these calli were transferred to the 100 combinations of regeneration media using various plant growth regulators; TDZ, IAA, 2ip, BAP, NAA, and zeatin. These calli derived from protoplast of three species of capsicum were, however, not differentiated into shoots.

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Taxoids from the Heartwood of Taxus baccata L. Growing in Turkey

  • Erdemoglu, Nurgun;Sener, Bilge
    • Natural Product Sciences
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    • v.6 no.2
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    • pp.96-101
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    • 2000
  • The ethanolic extract of the heartwood of Taxus baccata L. afforded six taxoids, belonging to the skeletally three different groups, $2{\alpha},\;5{\alpha},\;10{\beta}-triacetoxy-14{\beta}-(2-methyl)-butryloxy-4(20)$,11-taxadiene (1), taxusin (2), baccatin VI (3), baccatin III (4), $1{\beta}-hydroxybaccatin$ I (5) and taxol (6). The isolation and structure elucidation of three taxoids (3-5) have been presented in this study. This is the first report for the presence of these taxoids in Taxus baccata L. growing in Turkey.

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The proper concentrations of dextrose and lidocaine in regenerative injection therapy: in vitro study

  • Woo, Min Seok;Park, Jiyoung;Ok, Seong-Ho;Park, Miyeong;Sohn, Ju-Tae;Cho, Man Seok;Shin, Il-Woo;Kim, Yeon A
    • The Korean Journal of Pain
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    • v.34 no.1
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    • pp.19-26
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    • 2021
  • Background: Prolotherapy is a proliferation therapy as an alternative medicine. A combination of dextrose solution and lidocaine is usually used in prolotherapy. The concentrations of dextrose and lidocaine used in the clinical field are very high (dextrose 10%-25%, lidocaine 0.075%-1%). Several studies show about 1% dextrose and more than 0.2% lidocaine induced cell death in various cell types. We investigated the effects of low concentrations of dextrose and lidocaine in fibroblasts and suggest the optimal range of concentrations of dextrose and lidocaine in prolotherapy. Methods: Various concentrations of dextrose and lidocaine were treated in NIH-3T3. Viability was examined with trypan blue exclusion assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Migration assay was performed for measuring the motile activity. Extracellular signal-regulated kinase (Erk) activation and protein expression of collagen I and α-smooth muscle actin (α-SMA) were determined with western blot analysis. Results: The cell viability was decreased in concentrations of more than 5% dextrose and 0.1% lidocaine. However, in the concentrations 1% dextrose (D1) and 0.01% lidocaine (L0.01), fibroblasts proliferated mildly. The ability of migration in fibroblast was increased in the D1, L0.01, and D1 + L0.01 groups sequentially. D1 and L0.01 increased Erk activation and the expression of collagen I and α-SMA and D1 + L0.01 further increased. The inhibition of Erk activation suppressed fibroblast proliferation and the synthesis of collagen I. Conclusions: D1, L0.01, and the combination of D1 and L0.01 induced fibroblast proliferation and increased collagen I synthesis via Erk activation.

Studies on leptospiral antibody in canine and murine species (Leptospira 속균(屬菌)에 대한 개와 쥐의 혈중항체조사(血中抗體調査))

  • Ryu, E.P.;Suh, I.S.
    • Korean Journal of Veterinary Research
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    • v.11 no.1
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    • pp.41-47
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    • 1971
  • Investigation of Leptospira icterohaemorrhagiae and Leptospira canicola in dogs was carried out from May, 1970 to March, 1971 in Korea. A total of 943 random samples from 18 animal hospitals in 9 cities were collected and sent to Taipei and tested by the rapid microscopic agglutination test giving the following results. 1. The 943 samples gave 141 positives (14.95%). 2. Among the 18 animal hospitals a total 14 positives were found, with 30.0% being the highest and 9.1% the lowest reaction, comparing each hospital. 3. Monthly positive comparison gave the highest reaction for December (31.3%) the lowest August (5.8%). Seasonal comparison June, July and August was the lowest of all four seasons. 4. Of all the 913 cases examined, males were 505 (55.3%) and females 408 (44.7%), which gives only a difference of about 10% on the basis of sex. Comparing positive reaction cases on the basis of sex gave a large difference, of almost twice. Male${\cdots}$92: Female${\cdots}$47 5. Comparing 436 (46.9%) sick dogs against 492 (53.1%) healthy dogs gave 13.3 and 16.1% positive respectively. Thus indicating that some of the healthy dogs have inapparent infection. 6. Comparing on the basis of positive serum type L. icterohaemorrhagiae gave 133 (54.7%) and L. canicola 110 (45.3%) Generally both L. icterohaemorrhagiae and L. canicola produce common reaction with positive serum. 7. Comparing by age a total of 89 positive cases gave the following results. less than 1 year${\cdots}$49 (55.5%) from 1 year to 2 years${\cdots}$26 (29.2%) from 2 year to 3 years${\cdots}$12 (13.5%) over 3 years${\cdots}$5 8. Among 80 field rats collected at Suwon and tested by rapid microscopic agglutination test for L. icterohaemorrhagiae and L. canicola gave all negative reactions.

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Fabrication of a influenza A H1N1 sensor using ZnO nanostructure (산화아연 나노구조를 이용한 H1N1 인플루엔자 A 바이러스 센서 제작)

  • Jang, Yun-Seok;Park, Jung-Il;Nam, Yoon-Kyung;Pak, Jung-Ho
    • Proceedings of the KIEE Conference
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    • 2011.07a
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    • pp.1690-1691
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    • 2011
  • 본 논문에서는 H1N1 인플루엔자 A 바이러스(influenza A H1N1 virus) 검출을 위한 산화아연 나노구조(zinc oxide nano structure) 기반의 전기화학적 면역센서를 제작하고 그 특성을 분석하였다. H1N1 인플루엔자 A 바이러스는 빠른 전파 속도 때문에 정확하고 빠른 검출이 필요하다. 먼저, 2 $mm^2$의 표면적을 갖는 패턴된 금 전극 위에 열수방식(hydrothermal method)으로 성장시킨 산화아연 나노구조가 선택적으로 형성되도록 리프트-오프(lift-off) 방법을 사용하였다. 0.01 M phosphate buffered saline(pH 7.4)에서 2 ${\mu}g$/mL 농도의 1차 항체를 정전기력에 의해 산화아연 나노구조에 고정화한 후, 10 pg/mL ~ 5ng/mL 농도의 H1N1 항원을 적용하여 포획 항체에 결합시키고 HRP(horseradish peroxidase) 효소가 결합된 검출 항체를 항원에 결합시키는 샌드위치 ELISA법을 이용하였다. HRP와 반응하는 TMB(3,3', 5,5'-tetramethylbenzidine)와 과산화수소가 포함된 acetate buffered 용액(pH 5)을 전해질로 사용하고 순환전압전류 측정법(cyclic voltammetry)으로 센서의 특성을 분석하였다. 측정된 순환전압전류그래프(cyclic voltammogram)에서 H1N1 항원 농도 10 pg/mL ~ 5 ng/mL의 응답 전류는 276.47 ${\pm}$ 21.72 nA (평균 ${\pm}$ 표준편차, n=4) ~ 478.89 ${\pm}$ 6.21 nA로 측정되었고, logarithmic하게 증가하는 응답 전류 특성을 보였다.

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