• Title/Summary/Keyword: In vivo와 In vitro

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Selection and Characterization of Antagonistic Microorganisms for Biological Control of Acidovorax citrulli Causing Fruit Rot in Watermelon (수박에 과실썩음병을 유발하는 Acidovorax citrulli의 생물학적 방제를 위한 길항 미생물 선발과 특성 검정)

  • Kim, Ki Young;Park, Hyo Bin;Adhikari, Mahesh;Kim, Hyun Seung;Byeon, Eun Jeong;Lee, In Kyu;Lee, Youn Su
    • Research in Plant Disease
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    • v.28 no.2
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    • pp.69-81
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    • 2022
  • This study was performed to screen the efficacy of antagonistic bacterial isolates from various sources against the bacterial fruit blotch (BFB) causing pathogen (Acidovorax citrulli) in cucurbit crops. In addition, plant growth promoting traits of these antagonistic bacterial isolates were characterized. Two thousand seven hundred ninety-four microorganisms were isolated from the collected samples. Molecular identification revealed two A. citrulli out of 2,794 isolates. In vitro antagonistic results showed that, among the 28 antagonistic bacterial isolates, 24 and 14 bacterial isolates exhibited antagonism against HPP-3-3B and HPP-9-4B, respectively. Antagonistic and growth promotion characterization of the antagonistic bacterial isolates were further studied. Results suggested that, 4 antagonistic bacteria commonly showed both antagonism and growth promotion phenotypes. Moreover, 3 isolates possessed growth promoting activities. Overall results from this study suggests that BFB causing bacterial pathogen (A. citrulli) was suppressed in in vitro antagonism assay by antagonistic bacterial isolates. Furthermore, these antagonistic bacterial isolates possessed growth promotion and antagonistic enzyme production ability. Therefore, data from this study can provide useful basic data for the in vivo experiments which ultimately helps to develop the eco-friendly agricultural materials to control fruit rot disease in cucurbit crops in near future.

Study of Rat Mammary Epithelial Stem Cells In Vivo and In Vitro (생체 및 시험관에서 유선 상피 모세포의 분리와 동정)

  • Nam Deuk Kim;Kee-Joo Paik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.470-486
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    • 1995
  • Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

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Effects of Ethanol Extracts from Red Pepper (Capsicum annuum L.) Seeds on Cholesterol Adsorption Capacity and UDP-Glucuronyl Transferase Activity (고추씨 에탄올 추출물이 콜레스테롤 흡착능 및 UDP-glucuronyl transferase 활성에 미치는 영향)

  • Song, Won-Young;Kim, Yu-Na;Chun, Sung-Sik;Ku, Kyung-Hyung;Choi, Jeong-Hwa
    • Journal of Life Science
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    • v.21 no.6
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    • pp.829-837
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    • 2011
  • The purpose of this study was to investigate the effects of ethanol extracts from red pepper seeds on cholesterol adsorption capacity and UDP-glucuronyl transferase activity. In vitro cholesterol adsorption capacity of 2%, 5% and 10% ethanol extracts from red pepper seed groups were significantly higher than that of the control group. Sprague-Dawley strain male rats weighing $100{\pm}10$ g were randomly assigned to one normal diet N group and experimental groups fed high fat and high cholesterol diet, which were divided into HF (0.0%), HEA (0.1%), HEB (0.2%), and HEC (0.5%) groups according to the amount of ethanol extracts from red pepper seeds added to their basal diet. The body weight gain in the HF group was higher than that in the N group, and those in the HEA, HEB and HEC groups were lower than that in the HF group However, there were no statistically significant differences among the all the groups. The hepatic triglyceride and total cholesterol contents in the N group was significantly lower than that in the HF group, and those in the HEA, HEB and HEC groups were lower than that in the HF group. The hepatic UDP-glucuronyl transferase activity in the N group was lower than that of the HF group and those in the HEA, HEB and HEC groups were lower than that of the HF group. The serum total cholesterol and triglyceride contents of the HF group were significantly higher than that of the N group, and those of the HEA, HEB and HEC groups were lower than that of the HF group. The serum HDL-cholesterol contents in all groups supplemented with the ethanol extracts from red pepper seeds were significantly higher than that of the HF group. The serum LDL-cholesterol contents of the HF group were significantly higher than that of the N group, and those of the HEA, HEB and HEC groups were lower than that of the HF group. The fecal total cholesterol contents were significantly higher in the HF group compared to the N group, and those of the HEB and HEC groups were lower than that of the HF group. The fecal triglyceride contents in the N group was higher than that of the HF group, and those of the HEA, HEB and HEC groups were lower than that of the HF group. This study suggested that the ethanol extracts from red pepper seeds have powerful health benefits by the UDP-glucuronyl transferase activity and lipid metabolism.

Effects of a Blend of Prunus Mume Extract as an Alternative to Antibiotics on Growth Performance, Activity of Digestive Enzymes and Microflora Population in Broiler Chickens (항생제 대체 사료첨가제로서 매실추출 혼합물이 육계의 성장, 소화 효소 활성도 및 장내 미생물 균총에 미치는 영향)

  • Ko, Y.H.;Yang, H.Y.;Kang, S.Y.;Kim, E.S.;Jang, I.S.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.611-620
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    • 2007
  • The current study was designed to define whether a blend of prunus mume extract(25%) containing lactic acid(75%) and grape seed extract(10ppm) could affect in vitro antimicrobial activity and growth performance, intestinal microflora, plasma biochemical profiles and digestive enzymes activities in broiler chickens. In paper disc agar diffusion test, we clearly observed antimicrobial activity against E. coli in response to prunus mume extract or a blend of prunus mume extract. For in vivo test, a total of ninety six 3-d-old male broiler chicks were assigned to basal diet(CON), basal diet supplemented with antibiotics (ANTI) and 0.5% a blend of prunus mume extract(PRNUS) until 35 days of age. Throughout the entire experimental period(3-35 days), there were no differences in BW and FCR between the birds fed the basal diet with antibiotics and the diet supplemented with a blend of prunus mume. However, ANTI group showed a significant increase in BW and total gain compared to CON group. The weights of digestive organs such as the pancreas and mucosal tissues were not affected by dietary treatments. There was no difference in plasma levels of glucose, cholesterol, AST and ALT activity. However, triglyceride in plasma increased(P<0.05) in the birds fed the diet supplemented with 0.5% a blend of prunus mume extract compared to those fed antibiotics supplemented diet. The activities of pancreatic trypsin and amylase, and intestinal hydrolase including disaccharidase were not affected by dietary treatment. The colony forming units(CFU) of lactobacillus in the lower ileal-cecum of the birds fed the diet supplemented with a blend of prunus mume extract was significantly(P<0.05) higher than that of birds fed antibiotic supplemented diet without affecting the CFU of E. coli. In conclusion, the birds fed the diet supplemented a blend of prunus mume as an alternative to antibiotics showed a similar growth performance and an significant increase in lactobacillus population compared with the birds fed basal and antibiotics supplemented diets.

Effect of N-Acetylcysteine on the Supetoxide Release, Chemotaxis from the Neutrophils and Glutathione Level of Plasma and Neutrophils (N-Acetylcysteine이 호중구의 Superoxide, Chemotaxis 및 혈장과 호중구의 Glutathione에 미치는 영향)

  • Song, Jeong-Sup;Lee, Sook-Young;Moon, Hwa-Sik;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.475-483
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    • 1994
  • Background: N-acetylcysteine(ACE) is used both orally and intravenously in a variety of experimental pathologies resembling human disease states which exhibit endothelial toxicity as a result of oxidative stress, including acute pulmonary oxygen toxicity, septicemia and endotoxin shock. Despite these observations in vivo, it is not certain how this thiol drug produces its protective effects. ACE is a cysteine derivative which is able to direct1y react with oxygen radicals and may also act as a cysteine and glutathione(GSH) precursor following deacetylation. In this paper, we tried to know whether the therapeutic doses of ACE can modify the inflammatory function of the neutrophils and can increase the glutathione level of plasma in chronic obstructive pulmonary disease(COPD) patients. In addition, the effect of ACE to the purified neutrophil in terms of superoxide release and glutathione synthesis were observed. Method: Firstly, we gave 600mg of ACE for seven days and compare the release of superoxide, luminol-enhanced chemiluminescence from the neutrophils, neutrophil chemotaxis, and plasma GSH levels before and after ACE treatment in COPD patients. Secondly, we observed the dose dependent effect of ACE to the purified neutrophil's superoxide release and GSH levels in vitro. Results: 1) Usual oral therapeutic doses(600mg per day) of ACE for seven days did affect neither on the neutrophil's superoxide release, chemiluminescence, chemotaxis, nor on the plasma GSH concentration in the COPD patients. 2) ACE decreases the purified neutrophil's superoxide release and increase the GSH production in dose dependent fashion in vitro. Conclusion: Despite the fact that oral ACE treatment did not affect on the neutrophil's inflammatory function and plasma GSH concentration in COPD patients in usual therapeutic doses, it decreases the superoxide release and increases the GSH production from the isolated neutrophils in high molar concentrations. These findings suggest that to obtain an antioxidative effects of ACE, it might be needed to increase the daily dosage of ACE or therapeutic duration or change the route of adminisration in COPD patients.

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Protective Effect of a Herbal Preparation (HemoHIM) on the Self-Renewal Tissues and Immune System against γ-Irradiation (방사선에 대한 생약복합조성물(HemoHIM)의 재생조직 및 면역계 방호 · 회복촉진 효과)

  • Jo, Sung-Kee;Park, Hae-Ran;Jung, Uhee;Oh, Heon;Kim, Sung-Ho;Yee, Sung-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.805-813
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    • 2005
  • In our previous study, a novel herb mixture (HIM-I) of Angelim gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix was developed to protect the intestinal and immune systems and promote its recovery against radiation damage. In this study, a new herbal preparation (HemoHIM) with the high immune modulating activity was developed from HIM-I. HIM-I was fractionated into ethanol fraction (HIM-I-E) and polysaccharide fraction (HIM-I-P). And HemoHIM was prepared by adding HIM-I-P to HIM-I. The protective activities against $\gamma$ -irradiation were compared among HemoHIM, HIM-I and the fractions. HemoHIM and HIM-I significantly decreased the radiation-induced DNA damage in vitro, and scavenged hydroxyl radicals in a dose-dependent manner. HemoHIM showed similar activity to HIM-I. In vitro proliferation assay with mouse lymphocytes and bone marrow cells showed that HIM-I-P was remarkably higher than HIM-I and HIM-I-E in cell proliferating activity. HemoHIM showed higher activity than HIM-I and this might be associated with the higher polysaccharide content. The in vivo protective effects of HemoHIM and HIM-I were investigated in $\gamma$-irradiated mice. HemoHIM increased the surviving intestinal crypts to a similar extent compared with HIM-I. In contrast, HemoHIM appeared to be more effective than HIM-I in endogenous spleen colony formation assay. The recovery of white blood cells and lymphocytes in irradiated mice were significantly enhanced by the administration of HemoHIM. Also HemoHIM administration prolonged the survival of irradiated mice. These results showed that the novel herbal preparation, HemoHIM, effectively protected the self-renewal tissues and immune system, and promoted the survival of irradiated mice. Moreover, in comparison with HIM-I, HemoHIM maintained similar activity in the reduction of oxidative damage of self-renewal tissue but exhibited the higher activity in protection and proliferation of immune and hematopoietic cells. These results suggested that HemoHIM might be more effective than HIM-I in immune modulation as well as radioprotection.

Antioxidation Effect of Leg Bone Extracts and Enzyme Hydrolysates from Jeju Crossbred Horses (Jeju Native Horse×Thoroughbred) (제주 재래마와 더러브렛종과의 교잡종인 한라마 사골추출물과 효소분해물의 항산화 효과)

  • Kim, Dongwook;Pak, Jae-In;Chae, Hyun-Seok;Kim, Young-Boong;Jang, Aera
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1147-1154
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    • 2013
  • This study was conducted to evaluate the antioxidation activity of Jeju crossbred horse (Jeju native horse ${\times}$ thoroughbred) leg bone extracts (HLBE) and its enzyme hydrolysates by determination of 1,1-diphenyl-2picryl-hydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzo thiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity, ferric reducing/antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC). HLBE was extracted with hot water for 24 hr and lyophilized. The lyophilized HLBE was hydrolyzed using multifect PR6L (MP), pepsin (PS), and a pepsin and pancreatin mixture (PSPC) for 4 hr at 60, 50, and $50^{\circ}C$, respectively. The hydrolysates were separated by a molecular weight of 3 kDa more or less. When the yield of HLBE was 100%, the yield of hydrolysates less than 3 kDa of MP, PS, and PSPC was 10.86, 3.26, and 8.00%, respectively. Enzyme hydrolysates with low molecular weight less than 3 kDa in MP and PSPC showed significantly higher DPPH, ABTS radical scavenging activity, and ORAC compared to HLBE and its hydrolysates with more than 3 kDa. However, the FRAP of the hydrolysates less than 3 kDa in PS was significantly higher than in MP. These results suggest that low molecular weight enzyme hydrolysates less than 3 kDa have more powerful antioxidation activity, especially when they are hydrolyzed by MP and PSPC rather than PS. Therefore, low molecular weight enzyme hydrolysates of HLBE hydrolyzed with MP and PSPC have significant potential as antioxidants in the food industry. Further in vivo studies are required to support the antioxidation activities of the hydrolysates in vitro.

Evaluation on the Usefulness of Filter in Sentinel Lymphoscintigraphy Using $^{99m}Tc$-Phytate (Phytate를 이용한 감시림프절 검사 시 Filter의 유용성 평가)

  • Jeong, Yeong-Hwan;Seo, Han-Kyung;Shim, Cheol-Min;Lim, Seong-Dong;Han, Dong-Hyeon;Park, Yung-Sun;Kim, Dong-Yun
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.1
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    • pp.35-39
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    • 2010
  • Purpose: The aim of this study was to investigate distribution of particle size in phytate kit and compare filtered method with non-filtered method using 200 nm filter for sentinel lymphoscintigraphy (SLS). Materials and Methods: Five phytate kit of having the same available period was measured by particle size analyzer. For in-vivo experiment, $^{99m}Tc$-phytate was injected intradermally at both foot to perform lymphoscintigraphy. Imaging was acquired at 1hour after injection. Region of interest (ROI) was drawn in inguinal and background area for analysis. RAW 264.7 cells (Murine macrophage cell) were prepared for measurement of celluar uptake as a representative of macrophages. Paired t-test was performed using SPSS (SPSS Inc, USA) for statistical analysis. Results: The size of most particle in Techne phytate kit was distributed in 130~650 nm(90.5 %). In-vivo study, the ROI analysis showed similar result between filtered and non-filtered sample, and the numerical value of count/pixel were $58.3{\pm}5.97$ and $60.2{\pm}4.88$. In-vitro study, cellular uptake study also showed no difference between filtered and non-filtered sample by gamma counting. Conclusion: The present study demonstrates that there was no meaning of 200 nm filtered method for SLS using $^{99m}Tc$-phytate.

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Quantitative Analysis of Amino Acids of Paragonimus Westermani Treated with Praziquantel in Vivo and in Vitro (간흡충(肝吸蟲) 성충(成蟲)에 대한 Praziquantel의 시험관내 살충작용(殺蟲作用)에 의한 유리(遊離) 아미노산(酸)의 정량분석(定量分析))

  • Tak, Byung-Yun;Chung, Myung-Sook;Joo, Kyoung-Hwan;Rim, Han-Jong
    • Journal of agricultural medicine and community health
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    • v.16 no.2
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    • pp.134-140
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    • 1991
  • The amino acid constituents of Paragonimus westermani were very imperfectively known. Lee(1964) detected 9 amino acids in the tissue hydrolysates of P. westermani, and 13 amino acids were detected from the cyst content and body fluid constituents of P. ohirai, But, the quantity of amino acids in P. westermani is still unknown. In the present investigation 18 amino acids, the fundamental constituents of proteins, were quantitatively studied by high performance liquid chromatography. The results obtained were as follows : A total of 18 amino acids were recovered in protein hydrolysates of P. westermani obtained from cat. ; They were cystein, aspartic acid, glutamic acid, serine. glycine, histidine. arginine, threonine, alanine, proline, tyrosine, valine, methionine, isoleucine, leucine, phenylalanine, tryptophan and lysin. Among them, glutamic acid was the most abundant form and tryptophan, cystein, methionine, and histidine constitute minor portion of hydrolysates. Compared to the normal P. westermani, the volume of hydrolysates obtained from the praziquantel(PZQ) treated worm-0.lug PZQ/ml saline for 6 hours incubation, and $3{\times}25$mg/kg bwt${\times}$2days in vivo treatment was generally increased except tryptophan. A total of 17 free amino acids were identified and the volume was 174.18 umol/1 gm wet weight P. westermaini. Among them, glycine and alanine constitute 28% of total volume. No significant differences were observed in the material obtained from worm treated with PZQ. However, slight increase of serine, arginine and the slight diminution of glutamic acid and proline was observed.

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Estrogeicity of Genistein and Bisphenol A (콩류식품의 주성분인 Genistein과 식품포장재 및 용기에 사용되는 Bisphenol A의 에스트로젠 효과에 관한 연구)

  • 강경선;이영순;신광순
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.106-111
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    • 1998
  • This study has been focused on both estrogenic and proliferating activity of genistein (GEN) and bisphenol A (BPA). GEN and BPA enhance the proliferation of estrogen-dependent MCF-7 human breast cancer cells at concentrations as low as 100 nM of GEN and 8 ng/ml of BP A achieving similar effect to that of estradiol at 1 nM. Expression of the estrogen responsive gene, pS2 was also induced in MCF-7 cells by treatment with genistein at dose as low as 1 nM and BPA at dose as low as 4 ng/ml. Using 21 day-old ovariectomized nude mice, we examined end-bud formation and mammary gland development after treatment with bisphenol A or genistein. Compared with untreated control, mammary gland development and end-bud formation were significantly increased in mice fed genistein or bisphenol A (p<0.05). Taken together, it is concluded that GEN and BP A can act as an estrogen agonist resulting in cell proliferation and induction of the estrogen responsive pS2 gene in MCF-7 cells in vitro and in athymic mice in vivo, respectively. Therefore, it is suggested that GEN and BP A might modulate human endocrine system and these compounds might be considered as a endocrine modulator at the low levels of doses.

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