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Reduction of the Antigenicity of Whey Protein by Enzymatic Hydrolysis (효소가수분해에 의한 유청단백질의 항원성 저하)

  • Ha, Woel-Kyu;Juhn, Suk-Lak;Kim, Jung-Wan;Lee, Soo-Won;Lee, Jae-Young;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.26 no.1
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    • pp.74-80
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    • 1994
  • As a preliminary study about the reduction of the antigenicity of whey protein isolate(WPI) by treatment of chymotrypsin, trypsin, pancreatin, and protease from Aspergillus oryzae, the properties and antigenicities of whey protein hydrolysates(WPH) were investigated. When degrees of hydrolysis (DH) were measured by use of trinitrobenzensulfonic acid(TNBS), the DH of the WPH treated by pancreatin or protease from Aspergillus oryzae$(5.05{\sim}11.47)$ were much higher than those of the tryptic or chymotryptic WPH$(15.67{\sim}20.20)$. And the pretreatments of heat$(75^{\circ}C)$, 20 min and/or pepsin resulted in higher DH of WPH, generally. When the molecular distributions of the WPH were determined by high performance size exclusion chromatography(HPSEC), the ratios of polypeptides with molecular weight more than 10kDa ranged from 12% to 36%, and the average molecular weights and the average peptide lengths of the WPH were $4,252{\sim}9,132$ dalton and $38{\sim}83$ amino acids, respectively. And there was no bitter taste in all of the WPH. Results of SDS-PAGE showed that most of intact native proteins were eliminated by the enzymatic hydrolysis but there were a few bands of peptides larger than 14.2 kDa in some WPH. When antigenicity was assayed by competitive inhibition enzyme-linked immunosorbent assay(cELISA), monovalent antigenicity of WPH to rabbit anti-WPI antiserum were lowered to $10^{-1.7}-10^{-4.9}$ times and less by the enzymatic hydrolysis. And the pretreatments of heat and pepsin resulted in the lowest antigenicicy within a group of enzymatic hydrolysis, especially in case of the pancreatic hydrolysate(PDP) whose antigenicity was found almost to be removed.

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Effects of Seed Soaking of Kinetin on Alleviating Copper Toxicity during Germination in Rice (Kinetin 침종처리가 벼 발아중 구리 독성 경감에 미치는 영향)

  • 김상국;이상철;민기군;이승필;최부술
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.4
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    • pp.465-474
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    • 1996
  • The study was carried out to determine the proper concentration of plant growth regulator, kinetin on alleviating copper toxicity for two rice cultivars of seed germination. The results were as followings : Soaking treatment of kinetin 10$^{-3}$ M increased the germination rate of both cultivars, Ilpumbyeo and Hyangmibyeo 1 by 92% and 88% as compared with copper treatment (60ppm). But the soaking treatment effect of plant growth regulator, kinetin was not recognized at the kinetin 10$^{-4}$ M and 10$^{-5}$ M. Chlorophyll content of both rices was higher than that of Hyangmibyeo 1. Copper content of Ilpumbyeo was higher in leaf than in seed part. At the 3 days after treatment of copper 60ppm, both cultivars of treatment of kinetin 1O-3M showed the somewhat thin bands at the 35 and 40kDa compared with others. A new protein band pattern was only appeared to kinetin 1O-3M at approximately 54.4kDa(M. W) at the 7 days after treatment of copper 60ppm in llpumbyeo cultivar, SOD activity of copper 60ppm treatment increased in 3DAT, but there were no significant differences in 5 and 7DAT of two cultivars. Free proline contents of copper 60ppm treatment in llpumbyeo were remarkably increased about 4.996$\mu$M. In particular, free proline content of kinetin l0$^{-3}$ M in Ilpumbyeo was 5.008$\mu$M in 3DAT. In case of Hwangmibyeo 1, free proline content of copper 60ppm was 5.825$\mu$M compared with an untreated control showing 2.34l$\mu$M. The effects of kinetin treatment were recognized to promote the root growth and germination rate under copper toxicity(60ppm) condition in both cultivars.

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Greenhouse Environment and Growth of Green Pepper (Capsicum annuum L.) in Greenhouse Covered with CEM BIO Film (CEM BIO Film 피복시설의 환경특성과 풋고추 생육)

  • Chun, Hee;Kim, Kyung-Je;Kwon, Young-Sam;Kim, Hyun-Hwan;Lee, Si-Young
    • Journal of Bio-Environment Control
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    • v.9 no.3
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    • pp.161-165
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    • 2000
  • Spectroradiometric light transmittance from 300 to 1,100nm in the greenhouse covered with the CEM BIO polyethylene film was greater than that in the greenhouse covered with polyethylene film (control). As a whole, solar radiation transmittance into greenhouse was a half level, due to shades caused by double layer covering, frame and equipment. Net radiation energy emitted throughout surface of the greenhouse covered with CEM BIO polyethylene film was 5,424.5W.m$^{-2}$ , which was lower by 2.9% as compared to that of the greenhouse covered with polyethylene film. Photosynthetically active radiation from 400 to 700nm of the greenhouse covered with CEM BIO polyethylene film was 3,861.2W.m$^{-2}$ , which was higher by 3.8% as compared to hat of the greenhouse covered with polyethylene film. Accumulated minimum air temperature from Oct. 7, 1997 to Oct. 16, 1997 of the greenhouse covered with CEM BIO polyethylene film was 100.5$^{\circ}C$, which was higher by 2.5$^{\circ}C$ as compared to that of the greenhouse covered with polyethylene film. As results, height, stem diameter, leaf count, leaf area, fresh weight and dry weight of green pepper plants and canopy production structure measured at 30 days after transplanting were enhanced. Mean fruit weight n the greenhouse covered with CEM BIO polyethylene film was 11.28 g and 1.25 g greater as compared to that in the greenhouse covered with polyethylene film, due to increased fruit diameter and flesh thickness. Percent marketable fruits produced in the greenhouse covered with CEM BIO polyethylene film were 96.1%, and was greater by 2.7% thant that of the greenhouse covered with polyethylnee film due to decreased infection, sterility, severe curve and twisted fruits. The green pepper yield of the greenhouse covered with CEM BIO polyethylene film from Nov. 19, 1997 to Feb. 3, 1998 was greater by 974 kg per hectare than that of the greenhouse covered with polyethylene film, but the total fruit had no difference.

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THE EFFECTS OF DIETARY CONSISTENCY ON THE TRABECULAR BONE ARCHITECTURE IN GROWING MOUSE MANDIBULAR CONDYLE : A STUDY USING MICRO-CONFUTED TOMOGRAPHY (성장 중인 쥐에서 음식물의 경도가 하악 과두의 해면골에 미치는 영향 : 미세전산화 단층촬영을 이용한 연구)

  • Youn, Seok-Hee;Lee, Sang-Dae;Kim, Jung-Wook;Lee, Sang-Hoon;Hahn, Se-Hyun;Kim, Chong-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.2
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    • pp.228-235
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    • 2004
  • The development and proliferation of the mandibular condyle can be altered by changes in the biomechanical environment of the temporomandibular joint. The biomechanical loads were varied by feeding diets of different consistencies. The purpose of the present study was to determine whether changes of masticatory forces by feeding a soft diet can alter the trabecular bone morphology of the growing mouse mandibular condyle, by means of micro-computed tomography. Thirty-six female, 21 days old, C57BL/6 mice were randomly divided into two groups. Mice in the hard-diet control group were fed standard hard rodent pellets for 8 weeks. The soft-diet group mice were given soft ground diets for 8 weeks and their lower incisors were shortened by cutting with a wire cutter twice a week to reduce incision. After 8 weeks all animals were killed after they were weighed. Following sacrifice, the right mandibular condyle was removed. High spatial resolution tomography was done with a Skyscan Micro-CT 1072. Cross-sections were scanned and three-dimensional images were reconstructed from 2D sections. Morphometric and nonmetric parameters such as bone volume(BV), bone surface(BS), total volume(TV), bone volume fraction(BV/TV), surface to volume ratio(BS/BV), trabecular thickness(Tb. Th.), structure model index(SMI) and degree of anisotropy(DA) were directly determined by means of the software package at the micro-CT system. From directly determined indices the trabecular number(Tb. N.) and trabecular separation(Tb. Sp.) were calculated according to parallel plate model of Parfitt et al.. After micro-tomographic imaging, the samples were decalcified, dehydrated, embedded and sectioned for histological observation. The results were as follow: 1. The bone volume fraction, trabecular thickness(Tb. Th.) and trabecular number(Tb. N.) were significantly decreased in the soft-diet group compared with that of the control group (p<0.05). 2. The trabecular separation(Tb. Sp.) was significantly increased in the soft-diet group(p<0.05). 3. There was no significant differences in the surface to volume ratio(BS/BV), structure model index(SMI) and degree of anisotropy(DA) between the soft-diet group and hard-diet control group (p>0.05). 4. Histological sections showed that the thickness of the proliferative layer and total cartilage thickness were significantly reduced in the soft-diet group.

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A Study on Qian Yi(錢乙)'s Medical Though (전을(錢乙)의 의학사상(醫學思想)에 관(關)한 연구(硏究))

  • Oh, Jun Hwan;Kim, Ki Wook;Park, Hyun Kook
    • The Journal of Korean Medical History
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    • v.14 no.2
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    • pp.109-152
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    • 2001
  • Throughout this paper, I adjusted the study of 'Qian Yi'(錢乙)'s Medical Thought, and the following is the summary. 1. 'Qian Yi' wrote 'Xiao Er Yao Zheng Zhi Jue'("小兒藥證直訣", edited by 誾季忠), and there were 'Shang Han Lun Zhi Wei'("傷寒論指微"), 'Ying Ru Lun', however those are loss of the record. 2. Qian Yi's 'Zhi Jue'("直訣") was influenced by 'Lu Xin Jing', yet if we compare the quality of 'Sheng Li, Byeng Li, Bang Jae'(生理, 病理, 方劑), 'Lu Xin Jing' cannot be the foundation of 'Zhi Jue'. He took over 'Nei Jing, Shang Han Lun, Jin Gui Yao Lue, Shen Long Ben Cao Jing, Tai Ping Sheng Hui Fang'("內經", "傷寒論", "金?要略", "神膿本草經", "太平聖惠方") and put them together to the direct clinical experiences of pediatrics. 3. There is no reference regarding the difficulties of pediatric diagnosis and diseases in 'Huang Di Nei Jing'("黃帝內經") Before 'Bei Song'(北宋), regardless of the lack of data related to pediatric diseases, 'Qian Yi' established the pediatric system in 'Xiao Er Yao Zheng Zhi Jue' for the first time. 4. In his diagnosis of the pediatric diseases, he 'Si Zhen He Can'(四診合參), also considered in the eye exam seriously. In addition, he closely combined 'Wu Zang Bian Zheng'(五臟辨證), and diagnosis the pediatric diseases. 5. 'Wu Zang Bian Zheng', what Qian established method was based on 'Zheng Ti Guan'(整體觀) in 'Huang Di Nei Jing'. It was based on clinical experiences and established the perspectives of 'Tian Ren Xiang Ying'(天人相應). First of all, he pinpointed 'Zhu Zheng'(主證) clearly. Secondly, he pinpointed the relationships to symptoms and then, he distinguished a generic character of 'Xu, Shi, Han, Re'(虛, 實, 寒, 熱). Finally, he made an induction from genealogical pediatric physiology. 6. 'Qian Yi' took a serious view of 'Ban Zhen'(斑疹), the inadequate field in those days. At that time, he criticized on the habituation of the misuse of medication. He treated separately which 'Ji Jing'(急驚) as 'Liang Xie'(凉瀉) and 'Man Jing'(慢驚) as 'Wen Bu'(溫補). He proposed 'Cong Gan Zhu Feng, Xin Zhu Jing'(從肝主風, 心主驚) theory and formulated 'Jing Feng'(驚風) theory as well. 7. As an opponent of a tendency to misusage of medicine, 'Qian Yi' made out a prescription with pliant medicine. He emphasized on the treatment to 'Gong Bu Shang Zheng, Bu Bu Zhi Xie, Xiao Bu Jian Shi'(攻不傷正, 補不滯邪, 消補兼施) because he had so lucid demonstration to 'Xu Shi Han Re'(虛實寒熱) of the five viscera in the field of 'Bang Yak'(方藥). 8. There were no pediatrics schools at that time, however, the pediatrics was being made up gradually by 'Jin Yuan Si Da Jia'(金元四大家) who was influenced by 'Qian Yi'. He raised an objection to medical treatment using pliant medicine. 'Qian Yi' applied 'Qu Xia'(驅下) treatment using 'Han Liang'(寒凉) medicine. 'Han Liang Pai'(寒凉派) is greatly influenced by Qian. 'Chen Wen Zhong'(陳文中) had a great impact on 'Han Liang Pai' who used a 'Zao Shu Wen Bu'(燥熟溫補) medicine for treatment. Since 'Song Jin'(宋金), he had a tremendous influence on pediatrics treating patients in both 'Han Wen'(寒溫) ways. 9. 'Qian Yi' had an influence on his medical thoughts on future generations, especially to 'Wan Quan'(萬全) of 'Ming Dai', 'Wu Tang'(吳塘) of 'Qing Dai'(淸代) and 'Yun Shu Jie'(?樹珏) of 'Min Guo'(民國). 'Wan Quan' is an advocate of 'You Yu, Bu Zu Zhi Shuo'(有餘, 不足之說)of 'Xiao Er Wu Zang'(小兒五臟) that he revealed Qian's 'Wu Zang Bian Zheng'(五臟辨證). 'Wu Tang' disclosed Qian's 'Xiao Er Ti Zhi Shuo'(小兒體質說) and 'Xiao Er Ke'(小兒科)'s 'Yong Yao Lun'(用藥論), therefore, he uncovered pediatric physiological characteristics through the advocate of Qian's 'Zang Fu Rou Ruo, Ji Gu Nen Qie, Yi Xu Yi Shi, Yi Han Yi Re' (臟腑柔弱, 肌骨嫩怯, 易虛易實, 易寒易熱). 'Yun Shu Jie' developed intrinsic relationships among time, symptom and 'Tian Ren Xiang Ying Guan'(天人相應觀), What 'Qian Yi' stated about them. And also, he developed Qian's 'Di Huang Wan'(地黃丸), 'Xie Qing Wan'(瀉靑丸), 'Yi Huang San'(益黃散) clinical usages as well. 10. Regarding Qian's 'Wu Zang Xu Shi'(五臟虛實), it has an influence on 'Zhang Yuan Su'(張元素)'s 'Zang Fu Bing Ji Bian Zheng'(臟腑病機辨證). 'Di Huang Wan', 'Xie Qing Wan', 'Xie Xin Tang'(瀉心湯), 'Yi Huang San', 'Xie Huang San'(瀉黃散) are the standard prescription of 'Wu Zang Bu Xie'(五臟補瀉). It is under the influence of Qian's treatment. Besides, 'Qian Yi' took a serious view of 'Xiao Er'(小兒)'s 'Pi Wei'(脾胃). 'Qian Yi' had an impact on 'Li Dong Yuan'(李東垣) one of the member of 'Bu Tu Pai'(補土派). 'Di Huang Wan', which placed great importance on 'Bu Yi Shen Yin'(補益腎陰), had a great impact on 'Da Bu Yin Wan'(大補陰丸) and 'Jin Yuan Si Da Jia' as well. 11. In a theory of Qian's 'Wu Zang Bian Zheng', though it had been stated clearly in 'Wu Zang Bian Zheng', but he neglected in 'Liu Fu Bian Zheng'(六腑辨證). In prescription field, The problem with the medicine is that it is either toxic or mineral, therefore, we are not able to use those medicine in a clinical testing at the present time.

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Effects of Electrical Stimulation on the Biochemical Properties of Plaice, Paralichthys olivaceus, Sarcoplasmic Reticulum and Myofibrils (넙치 근소포체 및 근원섬유의 생화학적 특성에 미치는 전기자극의 영향)

  • KIM Tae-Jin;CHOI Young-Jun;KIM Dong-Su;CHO Young-Je
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.4
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    • pp.545-552
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    • 1998
  • This study was undertaken to clarify the effects of electrical stimulation on the biochemical properties of plaice sarcoplasmic reticulum and myofibrils at early period of death. The plaices were electrically stimulated (110V/60 Hz) In sea water bath for 15, 35, and 60 seconds, and killed instantly by spiking at the head. Killed samples were investigated for the changes in $Ca^{2+}$-ATPase activity of FSR (fragmented sarcoplasmic reticulum), LSR (light SR), HSR (heavy SR), and SDS-PAGE pattern of FSR. $Ca^{2+}$-ATPase activity of FSR increased until $45^{\circ}C$ and inactivated over $50^{\circ}C$. $Ca^{2+}$-ATPase activity of FSR remarkably decreased according to the duration of electrical stimulation. Myofibrillar $Mg^{2+}$-ATPase activity of electrically stimulated plaices in the presence of $Ca^{2+}$ was higher than that of sample instantly killed by spiking. $Mg^{2+}$-ATPase activity of myofibrils increased by electrical stimulation and the activity decreased during storage at $5^{\circ}C$. Myofibrillar $Mg^{2+}$-ATPase activity in sample killed by spiking was not affected by $Ca^{2+}$ ion. Myofibrillar $Mg^{2+}$-ATPase activity of electrically stimulated sample in the absen-re of $Ca^{2+}$ decreased during storage at $5^{\circ}C$, whit $Mg^{2+}$-ATPase activity in unstimulated sample did not show any change. $Ca^{2+}$-sensitivity of myofibrils showed no differences between electrically stimulated sample and sample killed by spiking, and the was no change during at $5^{\circ}C$.

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Comparative Study and Coloring Test for the Technique of Korean and Chinese Gold-painted porcelain (한(韓)·중(中) 화금자기(畵金磁器) 금채기법(金彩技法)에 대한 비교(比較) 조사(調査) 및 가채(加彩) 실험(實驗))

  • Hwang, Hyunsung
    • Conservation Science in Museum
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    • v.8
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    • pp.13-24
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    • 2007
  • The Department of Fine Art requested the conservation science team to examine the technique of painting with gold powder on the gold painted porcelains, which were made in Korea and China, among the items possessed by National Museum of Korea in order to publish it in the research paper collection of the Exchange Exhibition of Korea and China Porcelains. Among the items possessed by National Museum of Korea, such items were included as, an item of Koryo inlaid celadon (No. Gaesung 106), called the celadon with inlaid work (Korean name: Cheongjasanggam suhawonmun geumchaepyunho), which was decorated in the layer of glaze using gold power, an items of tea cup called "Temmku da wan" (No. Bongwan 10011), which was manufactured in the time of Song dynasty of Chinese history, an items of gold cup called "Siyuchohwamun hwageumwan" (No. Bongwan 2027), and an items of a porcelain called "Siyuwan" (No. Duksoo 3322). As the result of the examination, the stabilizing method of baking the decorated porcelains is similar, after they are painted with the pigment mixed with fine gold powder and the glaze solvent, but the treatment method of the colors is a little different among them. In other words, there is a big difference between them, for example, in the case of Koryo celadon, because the gold coloring was painted carefully one by one on the splendid decorations of inlaid works, while in the case of Chinese porcelains, butterflies or arabesque designs are decorated on the layer of the glaze in the porcelains of no decoration, using the stencil skill. According to the result of this examination, the part of the porcelain, whose layer of gold color was peeled off, could be restored, and as the result of the restoration, it was confirmed how beautiful and splendid the gold porcelain had been at the time of being manufactured.

Antioxidant and immunoregulatory effects of Korean Rhamnaceae (국내 자생 갈매나무과 식물의 항산화 및 면역반응조절 효과 탐색)

  • Eo, Hyun Ji;Kim, Da Som;Kang, Yeongyeong;Kim, Kiyoon;Park, Youngki;Park, Gwang Hun
    • Journal of Plant Biotechnology
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    • v.47 no.3
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    • pp.254-259
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    • 2020
  • We studied the antioxidative and immunoregulatory properties of 70% ethanol extracts from the leaves, branches, and fruit of 13 species of the Rhamnaceae collected during the bearing season (Berchemia berchemiifolia, Berchemia floribunda, Hovenia dulcis, Paliurus ramosissimus, Rhamnella franguloides, Rhamnus crenata, Rhamnus davurica, Rhamnus koraiensis, Rhamnus parvifolia, Rhamnus ussuriensis, Rhamnus yoshinoi, Sageretia thea, and Ziziphus ujube). Total phenolic and flavonoid content values were determined using calibration curves of gallic acid and rutin, respectively. ABTS and DPPH radical scavenging assays were performed to determine antioxidative potential. Anti-inflammatory effects were studied using the nitric oxide (NO) inhibition assay in LPS-induced RAW264.7 cells. Extracts of Berchemia berchemiifolia, Sageretia thea, and Ziziphus jujuba had the highest total phenol and flavonoid content values. Extracts of Berchemia berchemiifolia, Paliurus ramosissimus, Rhamnella franguloides, and Sageretia thea had the highest ABTS radical scavenging activities. Berchemia berchemiifolia and Sageretia thea had the highest DPPH radical scavenging activities. Furthermore, extracts of Berchemia berchemiifolia, Berchemia floribunda, Paliurus ramosissimus, Sageretia thea, and Ziziphus jujuba showed strong anti-inflammatory activity through inhibition of NO production. The results of this study suggest that Rhamnaceae extracts can be used as natural antioxidants and immunomodulators.

Repeated Intramuscular-dose Toxicity Test of Water-soluble Carthami Flos (WCF) Pharmacopuncture in Sprague-Dawley Rats

  • Choi, Yoo-Min;Jung, Da-Jung;Kim, Seok-Hee;Kim, Jong-Uk;Yook, Tae-Han
    • Journal of Pharmacopuncture
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    • v.18 no.1
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    • pp.36-43
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    • 2015
  • Objectives: Water-soluble carthami flos (WCF) is a new mixture of Carthami flos (CF) pharmacopuncture. We conducted a 4-week toxicity test of repeated intramuscular injections of WCF in Sprague-Dawley rats. Methods: Forty male and 40 female rats were divided into 4 groups of 10 male and 10 female SD rats: The control group received 0.5 mL/animal/day of normal saline whereas the three experimental groups received WCF at doses of 0.125, 0.25, and 0.5 mL/animal/day, respectively. For 4 weeks, the solutions were injected into the femoral muscle of the rats alternating from side to side. Clinical signs, body weights, and food consumption were observed; opthalmological examinations and urinalyses were performed. On day 29, blood samples were taken for hematological and clinical chemistry analyses. Then, necropsy was conducted in all animals to observe weights and external and histopathological changes in the bodily organs. All data were tested using a statistical analysis system (SAS). Results: No deaths were observed. Temporary irregular respiration was observed in male rats of the experimental group for the first 10 days. Body weights, food consumptions, opthalmological examinations, urinalyses, clinical chemistry analyses, organ weights and necropsy produced no findings with toxicological meaning. In the hematological analysis, delay of prothrombin time (PT) was observed in male rats of the 0.25- and the 0.5-mL/animal/day groups. In the histopathological test, a dose-dependent inflammatory cell infiltration into the fascia and panniculitis in perimuscular tissues was observed in all animals of the experimental groups. However, those symptoms were limited to local injection points. No toxicological meanings, except localized changes, were noted. Conclusion: WCF solution has no significant toxicological meaning, but does produce localized symptoms. No observed adverse effect level (NOAEL) of WCF in male and female rats is expected for doses over 0.5 mL/animal/day.

Effect of Aminoglycoside Antibiotics on in-Vitro Morphogenesis from Cultured Cells of Chrysanthemum and Tobacco

  • Teixeira da Silva, Jaime A.;Fukai, Seiichi
    • Journal of Plant Biotechnology
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    • v.6 no.1
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    • pp.25-37
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    • 2004
  • Successful genetic transformation of plants requires non-chimeric selection of transformed tissues and their subsequent regeneration. With rare exceptions, most transformation protocols still rely heavily on antibiotics for selecting transgenic cells that contain an antibiotic-degrading selectable marker gene. Here, the morphogenic capacity of in-vitro explants of chrysanthemnum and tobacco stems and leaves (control and transgenic) changed with the addition of aminoglycoside antibiotics (AAs), In a test of 6 AAs, phytotoxicity occurred at concentrations of 10 to 25 and 50 to 100$\mu\textrm{g}$ $mL^{-1}$ in chrysanthemum and tobacco explants, respectively. Light conditions as well as explant source and size also had significant effects. The use of transverse thin cell layers (tTCLs), in conjunction with high initial AA selection levels, supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow-cytometric analyses revealed no endodu-plication in chrysanthemum, even at high AA levels. However, this phenomenon was observed in tobacco calli(8C or more), even at low AA concentrations (i.e., 5 to 10 $\mu\textrm{g}$ mL$^{-1}$ ).