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생물반응기를 이용한 황근 부정근의 대량증식과 추출물의 항산화 및 미백 활성 평가
이종두,현호봉,현혜진,장은비,고민희,윤원종,함영민,정용환,최훤,오유진,오대주 한국자원식물학회 2022 한국자원식물학회지 Vol.35 No.4
Hibiscus hamabo Sieb. et Zucc. (yellow hibiscus) is a deciduous semi-shrub plant and mainly growing in Jeju Island. This is known the unique wild hibiscus genus and classified as an 2nd grade of endangered plant for Korean Red List. In previous studies, properties of germination, ecological, genetical and salt resistance have been reported. In this study, we investigated mass-proliferated adventitious root using bioreactor, antioxidant and whitening effects to conduct functional ingredients. Yellow hibiscus were collected from Gujwa, Jeju by prior permission and they were introduced by explant type and various medium composition after surface sterilization. As a result, seed response rates were evaluated at range of 51.17 ~51.83%, in terms of comprehensive efficiency of shoot and root formation. In the case of adventitious root propagation condition was confirmed in half strength Murashige and Skoog medium salts, 30 ㎎/L sucrose, and 2 ㎎/L indole-3-butyric acid for 8 weeks in 5,000 mL bioreactor. We also compared between relationship with biomass and secondary metabolites accumulation by total phenolics content, the flavonoid content, DPPH free radical scavenging activity and melanin content. The results indicated that adventitious root mass proliferation, antioxidant and whitening effect could develop value of the high-quality cosmeceutical ingredient and further metabolite studies 황근(Hibiscus hamabo Sieb. et Zucc., 노랑무궁화)은 주로제주도 전역과 전남 남부지역에 자생하는 우리나라 유일의 야생 무궁화속의 낙엽성 반관목식물이다. 최근 무분별한 자생지파괴에 따른 개체수 감소로 환경부 지정 한국적색목록 내 멸종위기야생생물 2급으로 지정되어 있다. 선행연구에서는 황근의종자발아, 생태 및 유전적 특성, 내염성 등이 보고되었다. 따라서 본 연구에서는 생물반응기를 이용하여 황근 부정근의 대량증식, 항산화 및 미백효과를 조사하여 기능성 원료 활용 평가를 실시하였다. 황근 종자 및 식물체는 사전 환경부 채취 허가를 받은 제주시 구좌읍 관내지역에서 채집하였으며, 표면살균 후 MS 등 배지 조성을 조절하여 기내 세포주로 도입하였다. 그 결과, 종자의 전반적인 반응율이 지상부 신초 발생 및 지하부 뿌리 발근 측면에서 51.17∼51.83%로 가장 효율적인 것으로 나타났다. 5,000 mL 생물반응기에서 황근 부정근의 증식에 유리한 배지조건의 경우, 1/2 MS (Murashige and Skoog, 1962), 30 g/L 자당(sucrose), 2 ㎎/L IBA (indole-3-butyric acid)에서 8주 동안 배양한 것임을 확인하였다. 또한 총페놀 화합물 함량, 총플라보노이드 함량, DPPH 자유 라디칼 소거능 및 멜라닌 함량 분석을 통해 바이오매스 및 2차대사산물 축적의 연관관계를 비교하였다. 본 연구를 통해 황근 부정근의 대량증식, 항산화 및 미백 효능은 고부가가치 향장품 산업의 고품질 소재개발과 연계하여 추가적인 생리활성물질 연구의 가능성을 시사한다.
이종두,김희중,봉정균,권수일 대한핵의학회 1999 핵의학 분자영상 Vol.33 No.1
Purpose: Preamplifier and amplifier are very important parts for developing a portable counting or imaging gamma probe. They can be used for analyzing pulses containing energy and position information for the emitted radiations. The commercial Nuclear Instrument Modules (NIMs) can be used for processing these pulses. However, it may be improper to use NIMs in developing a portable gamma probe, because of its size and high price. The purpose of this study was to develop both preamplifier and amplifier and measure their performance characteristics. Materials and Methodes: The preamplifier and amplifier were designed as a charge sensitive device and a capacitor resistor-rsistor capacitor (CR-RC) electronic circuit, respectively, and they were mounted on a print circuit board (PCB). We acquired and analyzed energy spectra for Tc-99m and Cs-137 using both PCB and NIMs. Multichannel analyzer (Accuspec/A, Caberra Industries Inc., Meriden Connecticut, U.S.A) and scintillation detectors (EP-047 (Bicron Saint-Gobain/Norton Industrial EP-047 (Ceramics Co., Ohio, U.S.A) with 2″×2″ NaI (T1) crystal and R1535 (Hamamatsu Photonics K.K., Electron Tube Center, Shizuoka-ken, Japan) with 1″×1″NaI (T1) crystal) were used for acquiring the energy spectra. Results: Using PCB, energy resolutions or EP-047 detectors for Tc-99m and Cs-137 were 12.92% and 5.01%, respectively, whereas R1535 showed 13.75% and 5.19% of energy resolution. Using the NIM devices, energy resolutions of EP-047 detector for Tc-99m and Cs-137 were measured as 14.6% and 7.58%, respectively. However, reliable energy spectrum of R1535 detector could not be acquired, since its photomultiplier tube (PMT) requires a specific type of preamplifier. Conclusion: We developed a special preamplifier and amplifier suitable for a small sized gamma probe that showed good energy resolutions independent of PMT types. The results indicate that the PCB can be used in developing both counting and imaging gamma probe.
HeLa세포배양을 이용한 콜레라균독소 및 항독소에 관한 연구
李鍾斗 대한감염학회 1975 감염 Vol.7 No.1
The present report concerns the toxin yield in the filtrates observed on the basis of cytopathic effect on HeLa cells in cultures, and immunogenic activities of cholera experimental vaccines consisting of both the toxic filtrates combined with the dead bacteria by mouse protection test. Cell free culture filtrates of Vibrio cholerae serotype Ogawa 41 were prepared by filtering through a 0.22U membrane filter peptone water which had 0.5∼5% peptone, pH6.5 or 7.8, and was cultivated at 29℃ for 17∼72 hours. Then, 17∼72 hour filtrates were inoculated on HeLa cells in monolayer cultures respectively and cytopathic effect was checked in 4 hours. Heated filtrates (56℃, 45 minutes) were also tested in this same way. In this way two kinds of toxins in the filtrates were found: heat labile toxin and heat stable toxin. These toxic filtrates were tested for mouse lethal activity, agglutinating, vibriocidal, and neutralizing antibody. Finally, experimental vaccines were prepared with the filtrates only and the filtrates plus dead cells in accordance with the conventional procedure for the preparation of cholera vaccine, and were tested for the protective activity against vibrio challenge by mouse protection test, compared with conventional vaccine. The results obtained are summarized as follows: 1) Heat labile toxin and heat stable toxin were found easily in cell-free culture filtrates of Vibrio cholerae on the basis of cytopathic effect on HeLa cells in culture. These toxins, which were also different in antigenicity, were neutralized specifically with their respective antiserum. Heat labile toxin was produced on culture at 29℃ in peptone water containing 0.5, or 1.0% peptone with nearly absence of heat stable toxin in 17 hour culture and the toxin appeared in the early stage of culture, the logarithmic phase. In contrast, heat stable toxin yield was greater on culture at 37℃ in peptone water containing above 3% peptone and the toxin appeared in the later stage of culture, the stationary phase. Activity of heat labile toxin was suppressed temporarily by the rabbit normal serum but heat stable toxin was not suppressed. 2) Toxicity of heat labile and stable toxic filtrates to mice increased in direct proportion to the peptone concentration in peptone water, incubation time and temperature, and heat stable toxic filtrate had a tendency to be more toxic than heat labile toxin. This tendency was similar to toxicity in vitro, based on cytopathic effects. 3) Agglutinins and vibriocidins were induced in rabbits at low and moderate level by both heat labile toxic filtrates and heat labile plus stable toxic filtrates respectively. The titers of agglutinins and vibriocidins were higher in heat labile toxic filtrates than heat stable toxic filtrates. 4) Experimental cholera vaccines consisting of dead cholera vibrio-containing toxic filtrates had more mouse protective activity than the conventional cholera vaccines but the cholera culture filtrates only, had similar protective activity to the cholera conventional vaccines diluted 100 times.