• Title/Summary/Keyword: intact cells

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Improvement of Peroxidase Productivity by Optimization of Medium Composition and Cell Inoculum Size in Suspension Cultures of Sweet Potato (Ipomoea batatas) (고구마(Ipomoea batatase)현탁배양에서 배지조성 및 세포접종량의 적정화에 의한 Pemxidase생산성 향상)

  • 곽상수;김수경;정경희;유순희;박일현;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.91-97
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    • 1994
  • To improve the productivity of peroxidase (POD) of cell line SP-47 derived from cell suspension cultures of sweet potato (Ipomoea batatas (L) Lam.cv White Star), we optimized culture conditions including the composition and concentration of plant growth regulators and carbon source, and the cell inoculum size. When one g (fr wt) of cells was inoculated into 50 mL TL medium supplemented with l mg/L 2,4-D and 30g/L sucrose in 300 mL Erlenmeyer flask at 25$^{\circ}C$ in the dark (100rpm), the POD activity per g cell dry wt was maximized to be about 6,800 units after 25 days of subculture, which was about 30 times higher than that of intact roots of horseradish plants grown in the greenhouse, but the cell growth was maximum after 15 days of subculture. The protein content per g cell dry wt maintained almost plateau and after 25 days of subculture decreased as culture Proceeded further whereas the POD specific activity (unit/mg protein) was about two times higher after subculture and continuously increased from 12 days to the end of cultures (40 days). The POD isozyme patterns showed almost the same regardless of cell growth stage, but some acidic isozymes were slightly increased after 25 days of subculture. These results indicate that POD activity in suspension cultures of sweet potato is closely associated with cell growth and stresses derived from cell culture renditions and medium depletion. Due to its high POD activity the SPL47cell line seems to be suitable for the mass production of POD.

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On Improvement of Garlic Productivity by Inactivation of Virus in Garlics (마늘 Virus 불활성화에 의한 생산성 향상에 관하여)

  • LEE Chang Un
    • Korean journal of applied entomology
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    • v.20 no.1 s.46
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    • pp.6-14
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    • 1981
  • The effect of heat or chemotherapeutant treatment on the mosaic virus infected garlic(Allium sativum L.) scales and that of chemotherapeutant added to the culture medium were summarized as following. The treatment of the virus infected garlic scales at $37\~57^{\circ}C$ for 35 days to one hour in water or in air shelved no effort of inactivating the virus. Although treatment of the garlic scales at $62\~72^{\circ}C$ for 90 to five minutes reduced the mosaic symptom on the leaves of the garlic plants grown after the heat treatment, it reduced the growth vigor of the plants so greatly that complete inactivation of the virus in garlics was not feasible. The mosaic symptom on the leaves of garlic plant was reduced when the infected garlic scales were grown after 24 hours soaking in $10\~50\;ppm$ Malachite Green, 2,4-Dichlorophnoxy Acetic Acid, or in $20\~100\;ppm$ Quinhydron. These chemotherapeutants, however, inhibited the growth of garlic plant at the high concentration. Garlic scales soaked in $10\~50\;ppm$ Naphthyl Acetic Acid showed the least mosaic symptom without its complete extiction on the garlic leaves. When incorporated into the modified Murashige-Skoog's medium, $0.5\~l.5\;ppm$ Naphthyl Acetic Acid could inactivate the mosaic virus in newy developed garlic plants showing no mosaic symptom on the leaves, no inclusion bodies and intact nuclei in the leaf tissue cells.

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Effects of Red-Koji Fermented Scutellariae Radix Extracts on Lipopolysaccharide-induced Rat Acute Lung Injury (홍국발효 황금이 Lipopolysaccharide 유발 급성 폐손상에 미치는 영향)

  • Kim, Koang Lok;Kwon, Kyoung Man;Yun, Yong Jae;Lee, Young Jun;Park, Dong Il;Kim, Jong Dae;Jung, Tae Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.6
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    • pp.874-885
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    • 2012
  • In the present study, the possibility of whether the pharmacological effects of Scutellariae Radix Aqueous Extracts(SR) were favorably changed by report that lipopolysaccharide(LPS)-induced rat acute lung injury was treated with Red-Koji(Monascus purpureus 12002) fermentation. Three different dosages of Red-Koji fermented SR extract(fSR), 125, 250 and 500 mg/kg were orally administered once a day for 28 days before LPS(Escherichia coli 0111:B4) treatments, and then 5 hours after LPS treatment(500 ${\mu}g$/head, intra trachea instillation), all rats were sacrificed. Changes in the body weights, lung weights, pulmonary transcapillary albumin transit, arterial gas parameters(pH, $PaO_2$ and $PaCO_2$) bronchoalveolar lavage fluid(BALF) protein, lactate dehydrogenase(LDH) and proinflammatory cytokine tumor necrosis factor-${\alpha}$(TNF-${\alpha}$), interleukin-$1{\beta}$(IL-$1{\beta}$) contents, total cell numbers, neutrophil and alveolar macrophage ratios, lung malondialdehyde(MDA), myeloperoxidase(MPO), proinflammatory cytokine TNF-${\alpha}$ and IL-$1{\beta}$ contents were observed with histopathology of the lung, changes on luminal surface of alveolus(LSA), thickness of alveolar septum, number of polymorphonuclear neutrophils(PMNs). As results of LPS-injection, dramatical increases in lung weights, pulmonary transcapillary albumin transit increases in $PaCO_2$, decreases in pH of arterial blood and $PaO_2$, increases of BALF protein, LDH, TNF-${\alpha}$ and IL-$1{\beta}$ contents, total cells, neutrophil and alveolar macrophage ratios, lung MDA, MPO, TNF-${\alpha}$ and IL-$1{\beta}$ contents increases were detected with decreases in LSA and increases of alveolar septum and PMNs numbers, respectively as compared with intact control. Especially fSR 125 mg/kg showed quite similar favorable effects on the LPS-induced acute lung injuries as compared with 60 mg/kg of ${\alpha}$-lipoic acid and 250 mg/kg of SR. The results suggest that over 125 mg/kg of fSR extracts showed favorable effects on the LPS-induced acute lung injury mediated by their antioxidant and anti-inflammatory effects. Moreover, increases of the pharmacological effects of SR on LPS-induced acute lung injury were observed by Red-Koji fermentation in this study, at least 2-fold higher.

Resistance to Reactive Oxygen Species and Antioxidant Activities of Some Strains of Lactic Acid Bacteria from the Mustard Leaf Kimchi (갓김치에서 분리된 유산균의 활성산소종에 대한 저항성과 항산화 활성)

  • Lim, Sung-Mee
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.375-382
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    • 2010
  • In present study, five strains of Lactobacillus acidophilus GK20, Lactobacillus brevis GK55, Lactobacillus paracasei GK74, Lactobacillus plantarum GK81, and Leuconostoc mesenteroides GK104 isolated from the mustard leaf kimchi were investigated for resistance to reactive oxygen species (ROS) and antioxidant activity. L. acidophilus GK20, L. brevis GK55, L. paracasei GK74, and L. plantarum GK81 were resistant to hydrogen peroxide (0.5 mM), showing a survival rate of 50% or more. In particular, L. acidophilus GK20 and L. paracasei GK74 were the most superoxide anions-resistant and L. paracasei GK74 and L. plantarum GK81 were most likely survive hydroxyl radicals. Meanwhile, the intracellular cell-free extract (ICFE) from L. plantarum GK81 exhibited significantly higher DPPH radical scavenging values ($96.4{\pm}2.8%$) than the intact cells (IC). The ICFE of L. plantarum GK81 showed the highest superoxide radical scavenging ability and chelating activity for $Fe^{2+}$ ions among the 5 lactic acid bacteria (LAB) tested, and IC and ICFE from L. plantarum GK81 demonstrated excellent reducing activity, which was higher than those of BHA and vitamin C as a positive control.

An Experimental Study on the Effectiveness of Microwave Hyperthermia Combined with Radiation on the Small and Large Intestine in rats (흰쥐의 장조직에 X-선 조사와 마이크로파 온열요법의 효과에 관한 실험적 연구)

  • Ahn, Kyung-Sook;Lee, Kyung-Ja;Rhee, Chung-Sik
    • Radiation Oncology Journal
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    • v.5 no.2
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    • pp.83-95
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    • 1987
  • The synergistic effect of combining radiation therapy and hyperthermia kills significantly more cells than using either modality alone. The reason for enhanced cell killing from the combined treatment is that the two modalities are complementary. For histopathological exmination, 102 rats were divided into 4 groups as hyperthermia, radiation, hyperthermia combined with radiation and normal control groups. The effect of prior irradiation (6-15 Gy of X-ray) on the response of small and large bowel of rats to $40^{\circ}C-44^{\circ}C$ (for 30 minutes) microwave (2450 MHz) hyperthermia was investigated. The musculature of the small and large intestine remained intact and the circumference of the histological sections were not significantly altered by the heated at $43^{\circ}C$ for 30 minutes. Thermal enhancement ratios of normal tissue is 1.0 Thermal enhancement ratio was not increased in combination therapy by evaluation of histopathologic changes in small and large intestine.

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Detection Characteristics of TL, ESR and DNA Comet for Irradiated Peanuts by Origins (TL, ESR및 DNA Comet분석에 의한 원산지별 땅콩의 방사선 조사 검지 특성)

  • 이은영;정재영;조덕조;권중호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.6
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    • pp.1076-1081
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    • 2001
  • Gamma-irradiated peanuts, Korean and Chinese origins, were investigated on detection properties by thermoluminescence (TL), electron spin resonance (ESR), and DNA comet assay (single cell gel electrophoresis). TL measurement showed that the non-irradiated sample revealed a glow curve with low intensity at about 25$0^{\circ}C$, while the irradiated samples showed higher intensity around at 18$0^{\circ}C$. TL ratio (TL$_1$/TL$_2$) of area for TL$_1$ glow curve to TL$_2$ was below 0.05 for the non-irradiated sample and 0.2 or more for the irradiated ones, thus identifying each other. ESR spectroscopy for the irradiated peanuts using outer skin showed negligible signals induced by irradiation, indicating ESR is little applicable to the detection of irradiated peanuts. In DNA comet assay, the non-sample had no or very short tails, whereas the irradiated samples revealed the cells with long tails. Significance in the increase of their lengths depending on irradiation dose (r=0.761/Korean, r=0.768/Chinese) was also found. There was no remarkable difference in detection properties by origins of samples in all determinations, It is concluded that TL analysis or DNA comet assay is suitable for detection of irradiated peanuts and a combined method is recommendable for enhancing the reliability of detection results.

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Cloning and Functional Analysis of Gene Coding for S-Adenosyl-L-Methionine Synthetase from Streptomyces natalensis (Streptomyces natalensis로부터 S-adenosyl-L-methionine synthetase 유전자의 클로닝 및 기능분석)

  • Yoo, Dong-Min;Hwang, Yong-Il;Choi, Sun-Uk
    • Journal of Life Science
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    • v.21 no.1
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    • pp.96-101
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    • 2011
  • S-Adenosyl-L-methionine synthtase (SAM-s) catalyzes the biosynthesis of SAM from ATP and L-methionine. SAM plays important roles in the primary and secondary metabolism of cells. A metK encoding a SAM-s was searched from Streptomyces natalensis producing natamycin, a predominantly a strong antifungal agent, inhibiting the growth of both yeasts and molds and preventing the formation of aflatoxin in filamentous fungi. To obtain the metK of S. natalensis, PCR using primers designed from the two highly conserved regions for metK genes of Streptomyces strains was carried out, and an intact 1.2-kb metK gene of S. natalensis was cloned by genomic Southern hybridization with PCR product as a probe. To identify the function of the cloned metK gene, it was inserted into pSET152ET for its high expression in the Streptomyces strain, and then introduced into S. lividans TK24 as a host by transconjugation using E. coli ET12567(pUZ8002). The high expression of metK in S. lividans TK24 induced actinorhodin production on R5 solid medium, and its amount in R4 liquid medium was 10-fold higher than that by exconjugant including only pSET152ET.

Effects of Lonicerae Flos Extracts on LPS-induced Acute Lung Injury (금은화가 LPS로 유발된 급성 폐 손상에 미치는 영향)

  • Yi, Chang-Geon;Choi, Hae-Yun;Park, Mee-Yeon;Kim, Jong-Dae
    • Journal of Society of Preventive Korean Medicine
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    • v.15 no.1
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    • pp.49-69
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    • 2011
  • Objective : The object of this study was to observe the effects of Lonicerae Flos (LF) aqueous extracts on lipopolysaccharide (LPS)-induced rat acute lung injury. Method : Five different dosages of LF extracts were orally administered once a day for 28 days before LPS treatments, and then all rats were sacrificed after 5 hour-treatment of LPS. Eight groups of 16 rats each were used in the present study. The following parameters caused by LPS treatment were observed ; body weights, lung weights, pulmonary transcapillary albumin transit, arterial gas parameters (pH, $PaO_2$ and $PaCO_2$) bronchoalveolar lavage fluid (BALF) protein lactate dehydrogenase (LDH), and proinflammatory cytokines tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$) contents, total cell numbers, neutrophil and alveolar macrophage ratios, lung malondialdehyde (MDA), myeloperoxidase (MPO), proinflammatory cytokines TNF-${\alpha}$ and IL-$1{\beta}$ contents. In addition, the histopathologic changes were observed in the lung in terms of luminal surface of alveolus, thickness of alveolar septum, number of polymorphonuclear neutrophils. Result : As results of LPS-injection, dramatical increases in lung weights, pulmonary transcapillary albumin transit increases, increases in $PaCO_2$, decreases in pH of arterial blood and $PaO_2$, increases of BALF protein, LDH, TNF-${\alpha}$ and IL-$1{\beta}$ contents, total cells, neutrophil and alveolar macrophage ratios, TNF-${\alpha}$ and IL-$1{\beta}$ contents increases were detected with decreases in LSA and increases of alveolar septum and PMNs numbers, respectively as compared with intact control. These are means that acute lung injuries (resembling acute respiratory distress syndrome) are induced by treatment of LPS mediated by inflammatory responses, oxidative stress and related lipid peroxidation in the present study. However, these LPS-induced acute lung injuries were inhibited by 28 days continuous pretreatment of 250 and 500mg/kg of LF extracts. Because of lower three dosages of LF treated groups, 31.25 and 62.5 and 125mg/kg did not showed any favorable effects as compared with LPS control, the effective dosages of LF in LPS-induced acute lung injuries in the present study, is considered as about 125mg/kg. The effects of 250mg/kg of LF extracts showed almost similar effects with ${\alpha}$-lipoic acid 60mg/kg in preventing LPS-induced acute lung injuries. Conclusion : It seems that LF play a role in protecting the acute respiratory distress syndrome caused by LPS.

Effects of Jaeumkanghwa-tang on the Rat Hypothyroidism Induced by Propylthiouracil (PTU) (자음강화탕(滋陰降火湯)이 Propylthiouracil(PTU)로 유발된 Rat의 갑상선기능저하증에 미치는 영향)

  • Kim, Seok-Jung;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.27 no.1
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    • pp.41-64
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    • 2014
  • Objectives: This study was to evaluate the effect of Jaeumkanghwa-tang (JEKHT) on the propylthiouracil (PTU)-induced rat hypothyroidism. Methods: Six groups, each of 8 rats per group were used in the present study - intact vehicle control, PTU control, Levothyroxine ($LT_4$), JEKHT 500, 250 and 125 mg/kg treated groups. JEKHT were administered once a day for 42 days as an oral dose of 500, 250 and 125 mg/kg, and hypothyroidism was induced by daily subcutaneous treatment of PTU 10 mg/kg for 28 days. The changes on the body and organ weight, serum hormone and lipid profiles, liver and testis antioxidant defense factors were observed with histopathology of organs. Results were compared with $LT_4$ 0.5 mg/kg intraperitoneally treated rats in this experiment. Results: PTU treatment, marked decrease of body weight, increases of thyroid weight, decreases of liver, testis, epididymis and prostate weights, decreases of serum Tri-iodothyronine ($T_3$), and Thyroxine ($T_4$) level with increase of serum Thyroid-stimulating hormone (TSH) level, decreases of serum testosterone and dihydrotestosterone (DHT) level with increases of serum Follicular stimulating hormone (FSH) level, increases of serum High density lipoprotein (HDL), decrease of triglyceride content, increase of serum Aspartate aminotransferase (AST) level, decreases of liver and testis antioxidant defense factors were observed. In addition, marked hyperplasia of follicular cells with decreases of follicular colloid contents and diameters was additionally demonstrated with the decrease of hepatocyte numbers per unit area due to hypertrophy of hepatocytes related to lipid droplet depositions, increase of a/oligospermatic epididymal tubules with epididymal atrophic changes, seminiferous tubular atrophy with decrease of stage I~II seminiferous tubules in testis, prostate tubular atrophic changes at histopathological inspections. However, these PTU induced hypothyroidism and related hepatic and male reproductive organ damages were favorably and dose-dependently inhibited by treatment of JEKHT 500, 250 and 125 mg/kg, and JEKHT also effectively regulated the PTU-induced abnormal antioxidant defense factor changes in the both liver and testis. Conclusions: JEKHT 500, 250 and 125 mg/kg dose-dependently inhibited PTU-induced hypothyroidism and related liver and male reproductive organ damages in rats.

Ameliorating Effects of Nokyongdaebo-tang on Experimental Subacute Hemorrhagic Anemia in Rats (녹용대보탕 열수 추출물의 실험적으로 유발된 랫트 아급성 출혈성 빈혈에 대한 효과)

  • Kim, Jung-Ah;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.3
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    • pp.1-19
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    • 2017
  • Objectives: The object of this study is to observe the possible ameliorating effects of Nokyongdaebo-tang (NYDBT) on the experimental subacute hemorrhagic anemia (SHA) in rats. Methods: In the present study, SHA in rats was induced by exsanguinations from orbital plexus, and ameliorating effects of NYDBT was observed based on the changes of body and hematopoietic organ (spleen, liver and femur) weights, red blood cell (RBC) related hematological values, smear cytology, histopathological changes and immunohistochemistrical analysis of hematopoietic stem cells in the femur bone marrow, liver and spleen. In addition, the gastrointestinal motility and the surface mucosa thicknesses of remnant fecal pellets in the colon lumen, mucosa thicknesses and the mucous producing cell numbers in the colonic mucosa were analyzed to observe the digestive disorders, especially on the constipation, the major discomfort problems in iron supplement. Results: SHA related abnormal anemic signs were markedly and dose-dependently inhibited by oral administration of NYDBT 500, 250 and 125 mg/kg in a condition of this experiment. In addition, no meaningful changes on the gastrointestinal motilities and mucous component on the colon and remnant feces were noticed in all three different dosages of NYDBT treated rats as compared with intact vehicle and SHA control rats in this study. Conclusions: It, therefore, is expected that NYDBT will be promising as a novel alternative hematopoietic and therapeutic agent for anemia.