• Title/Summary/Keyword: Culture Conditions

Search Result 3,834, Processing Time 0.042 seconds

The Effect of Epigallocatechin-3-gallate on HIF-1 α and VEGF in Human Lung Cancer Cell Line (비소세포폐암주에서 저산소상태에 의해 유발된 HIFa-1 α와 VEGF의 발현증가에 미치는 Epigallocatechin-3-gallate의 억제 효과)

  • Song, Joo Han;Jeon, Eun Joo;Kwak, Hee Won;Lee, Hye Min;Cho, Sung Gun;Kang, Hyung Koo;Park, Sung Woon;Lee, Jae Hee;Lee, Byung Ook;Jung, Jae Woo;Choi, Jae Cheol;Shin, Jong Wook;Kim, Ki Jeong;Kim, Jae-Yeol;Park, In Won;Choi, Byoung Whui
    • Tuberculosis and Respiratory Diseases
    • /
    • v.66 no.3
    • /
    • pp.178-185
    • /
    • 2009
  • Background: Epigallocatechin-3-gallate (EGCG) is the major catechin in green tea, and has shown antiproliferative, antiangiogenic, antimetastatic and cell cycle pertubation activity in various tumor models. Hypoxia can be induced because angiogenesis is insufficient for highly proliferating cancer. Hypoxia-inducible factor-1$\alpha$ (HIF-1$\alpha$) and its downstream target, vascular endothelial growth factor (VEGF), are important for angiogenesis, tumor growth and metastasis. The aim of this study was to determine how hypoxia could cause changes in the cellular phenomena and microenvironment in a non-small cell culture system and to examine the effects of EGCG on a HIF-1$\alpha$ and VEGF in A549 cell line. Methods: A549 cells, a non-small cell lung cancer cell line, were cultured with DMEM and 10% fetal bovine serum. A decrease in oxygen tension was induced using a hypoxia microchamber and a $CO_2-N_2$ gas mixture. Gas analysis and a MTT assay were performed. The A549 cells were treated with EGCG (0, 12.5, 25, 50 ${\mu}mol/L$), and then examined by real-time-PCR analysis of HIF-1$\alpha$, VEGF, and $\beta$-actin mRNA. Results: Hypoxia reduced the proliferation of A549 cells from normoxic conditions. EGCG inhibited HIF-1$\alpha$ transcription in A549 cells in a dose-dependent manner. Compared to HIF-1$\alpha$, VEGF was not inhibited by EGCG. Conclusion: HIF-1$\alpha$ can be inhibited by EGCG. This suggests that targeting HIF-1$\alpha$ with a EGCG treatment may have therapeutic potential in non-small cell lung cancers.

Effects Of Cultured Bone Cell On The Regeneration Of Alveolar Bone (배양골세포 이식이 치조골재생에 미치는 영향)

  • Jeong, Soon-Joon;Herr, Yeek;Park, Joon-Bong;Lee, Man-Sup;Kwon, Young-Hyuk
    • Journal of Periodontal and Implant Science
    • /
    • v.26 no.1
    • /
    • pp.1-26
    • /
    • 1996
  • This study was performed to estimate the effects of cultured bone cell inoculated on porous type hydroxyaptite for the regeneration of the artificial alveolar bone defect. In this experiment 3 beagle dogs were used, and each of them were divided into right and left mandible. Every surgical intervention were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). To reduce the gingival bleeding during surgery, operative site was injected with Lidocaine hydrochloride(l:80,000 Epinephrine) as local anesthesia. After surgery experimental animal were feeded with soft dietl Mighty dog, Frisies Co., U.S.A.) for 1 weeks to avoid irritaion to soft tissue by food. 2 months before surgery both side of mandibular 1st premolar were extracted and bone chips from mandibular body were obtained from all animals. Bone cells were cultured from bone chips obtained from mandible with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Porous type hydroxyapatite were immerse into the high concentrated cell suspension solution, and put 4 hours for attachin the cells on the surface of hydroxyapatite. Graft material were inserted on the artificial bone defect after 3 days of culture. Before insertion of cellinoculated graft material, scanning electronic microscopic observation were performed to confirm the attachment and spreading of cell on the hydroxyapatite surface. 3 artificial bone defects were made with bone trephine drill on the both side of mandible of the experimental animal. First defect was designed without insertion of graft material as negative control, second was filled with porous replamineform hydroxyapatite inoculated with cultured bone marrow cells as expermiental site, and third was filled with graft materials only as positive control. The size of every artificial bone defect was 3mm in diameter and 3mm in depth. After the every surgical intervention of animals, oral hygiene program were performed with 1.0% chlorhexidine digluconate. All of the animals were sacrificed at 2, 4, 6 weeks after surgery. For obtaining histological section, tissus were fixed in 10% Buffered formalin and decalcified with Planko - Rycho Solution for 72hr. Tissue embeding was performed in paraffin and cut parallel to the surface of mandibular body. Section in 8um thickness of tissue was done and stained with Hematoxylin - Eosin. All the specimens were observed under the light microscopy. The following results were obtained : 1. In the case of control site which has no graft material, less inflammatory cell infiltration and rapid new bone forming tendency were revealed compared with experimental groups. But bone surface were observed depression pattern on defect area because of soft tissue invasion into the artificial bone defect during the experimental period. 2. In the porous hydroxyapatite only group, inflammatory cell infiltration was prominet and dense connective tissue were encapsulated around grafted materials. osteoblastic activity in the early stage after surgery was low to compared with grafted with bone cells. 3. In the case of porous hydroxyapatite inoculated with bone cell, less inflammatory cell infiltration and rapid new bone formation activity was revealed than hydroxyapatite only group. Active new bone formation were observed in the early stage of control group. 4. The origin of new bone forming was revealed not from the center of defected area but from the surface of preexisting bony wall on every specimen. 5. In this experiment, osteoclastic cell was not found around grafted materials, and fibrovascular invasion into regions with no noticeable foreign body reaction. Conclusively, the cultured bone cell inoculated onto the porous hydroxyapatite may have an important role of regeneration of artificial bone defects of alveolar bone.

  • PDF

ANTIMICROBIAL ACTIVITY OF ARTEMISIA SPECIES AGAINST CLINICALLY ISOLATED STREPTOCOCCUS MUTANS (임상 분리된 Streptococcus mutans에 대한 Artemisia species의 항균 활성도)

  • Seo, Jung-Ah;Kim, Jae-Gon;Kim, Mi-Ah;Baik, Byeong-Ju;Yang, Yeon-Mi;Jeong, Jin-Woo
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.36 no.4
    • /
    • pp.505-513
    • /
    • 2009
  • Streptococcus mutans plays a major role in the formation of dental plaque and it is considered one of the important pathogens in the development of dental caries. Established dental plaque can be more resistant to antimicrobial agents and offers nutrient rich and relatively stable cariogenic conditions for bacteria. Thus bacteria growing in dental plaque have strong resistance to antimicrobial agents and they are not removed easily by the flow of saliva. Many researchers have been performed using natural materials, especially herbal extracts to prevent dental plaque. However, the strains of mutans streptococci used in the researches were not from Koreans. Therefore, it would be necessary to evaluate the antimicrobial activity of herbal extracts against clinical isolates of Streptococcus mutans isolated from saliva of Koreans living in Jeollabuk-do. For this study four clinical isolates were isolated from saliva samples of seventeen Korean people to investigate whether essential oils of Artemisia spp. have antimicrobial activity against them including four reference strains of Streptococcus mutans. Minimum inhibitory concentrations, Minimum bactericidal concentrations and time kill studies were performed and the results showed that Artemisia lavandulaefolia DC, Artemisia scoparia, and Artemisia capillaries have antimicrobial activity against the Streptococcus mutans clinical isolates and reference strains.

  • PDF

Expression of Sodium/iodide Symporter Transgene in Neural Stem Cells (신경줄기세포(HB1.F3)에서 나트륨옥소 공동수송체 도입유전자 발현)

  • Kim, Yun-Hui;Lee, Dong-Soo;Kang, Joo-Hyun;Lee, Yong-Jin;Chung, June-Key;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
    • /
    • v.38 no.1
    • /
    • pp.99-108
    • /
    • 2004
  • Purpose: The ability to noninvasively track the migration of neural progenitor cells would have significant clinical and research implications. We generated stably transfected F3 human neural progenitor cells with human sodium/iodide symporter (hNIS) for noninvasively tracking F3. In this study, the expression patterns of hNIS gene in F3-NIS were examined according to the cultured time and the epigenetic modulation. Materials and Methods: F3 human neural stem cells had been obtained from Dr. Seung U. Kim (Ajou University, Suwon, Korea). hNIS and hygromycin resistance gene were linked with IRES (Internal Ribosome Entry Site) under control of CMV promoter. This construct was transfected to F3 with Liposome. To investigate the restoration of hNIS gene expression in F3-NIS, cells were treated with demethylating agent (5-Azacytidine) and Histone deacetylase inhibitor (Trichostatin A: TSA). The expression of hNIS was measured by I-125 uptake assay and RT-PCR analysis. Results: The iodide uptake of the F3-NIS was higher 12.86 times than F3 cell line. According to the cell passage number, hNIS expression in F3-NIS gradually diminished. After treatment of 5-Azacytidine and TSA with serial doses (up to $20{\mu}M$, up to 62.5nM, respectively) for 24 hours, I-125 uptake and mRNA of hNIS in F3-NIS were increased. Conclusion: These results suggest that hNIS transfected F3 might undergo a change in its biological characters by cell passage. Therefore, the gene ex[ressopm of exogenous gene transferred human stem cell might be affected to the epigenetic modulation such as promoter methylation and Histone deacetylation and to the cell culture conditions.

Isolation and Characterization of Bacillus Strain as a Potential Biocontrol Agent (환경친화적 미생물농약으로서의 잠재성을 가진 세균의 분리 및 특성)

  • Lee, Ye-Ram;Lee, Sang-Mee;Jang, Eun-Young;Hong, Chang-Oh;Kim, Keun-Ki;Park, Hyean-Cheal;g Lee, Sang-Mon;Kim, Young-Gyun;Son, Hong-Joo
    • Journal of Life Science
    • /
    • v.25 no.12
    • /
    • pp.1408-1414
    • /
    • 2015
  • In this study, to retain a stable bacterial inoculant, Bacillus strains showing antifungal activity were screened. The improved production, antifungal mechanism, and stability of the antifungal metabolite by a selected strain, AF4, a potent antagonist against phytopathogenic Botrytis cinerea, were also investigated. The AF4 strain was isolated from rhizospheric soil of hot pepper and identified as Bacillus subtilis by phenotypic characters and 16S rRNA gene analysis. Strain AF4 did not produce antifungal activity in the absence of a nitrogen source and produced antifungal activity at a broad range of temperatures (25-40℃) and pH (7-10). Optimal carbon and nitrogen sources for the production of antifungal activity were glycerol and casein, respectively. Under improved conditions, the maximum antifungal activity was 140±3 AU/ml, which was higher than in the basal medium. Photomicrographs of strain AF4-treated B. cinerea showed morphological abnormalities of fungal mycelia, demonstrating the role of the antifungal metabolite. The B. subtilis AF4 culture exhibited broad antifungal activity against several phytopathogenic fungi. The antifungal activity was heat-, pH-, solvent-, and protease-stable, indicating its nonproteinous nature. These results suggest that B. subtilis AF4 is a potential candidate for the control of phytopathogenic fungi-derived plant diseases.

Biodegradation of Phenol by Comamonas testosteroni DWB-1-8 Isolated from the Activated Sludge of Textile Wastewater (섬유 폐수 활성 슬러지에서 분리한 Comamonas testosteroni의 생물학적 페놀 분해)

  • Kwon, Hae Jun;Choi, Doo Ho;Kim, Mi Gyeong;Kim, Dong-Hyun;Kim, Young Guk;Yoon, Hyeokjun;Kim, Jong-Guk
    • Journal of Life Science
    • /
    • v.30 no.2
    • /
    • pp.156-161
    • /
    • 2020
  • Since industrialization, the production and utilization of various chemicals has contributed to improving the quality of our lives, but the subsequent discharge of massive waste is inevitable, and environmental pollution is becoming more serious every day. Exposure to chemicals as a result of environmental pollution is having a negative effect on human health and the ecosystem, and cleaning up the polluted environment that can affect our lives is a very important issue. Toxic aromatic compounds have been detected frequently in soil, groundwater, and wastewater because of the extensive use of oil products, and phenol, which is used to produce synthetic resins, textiles, and dyes, is one of the major pollutants, along with insecticides and preservatives. Phenol can cause dyspnea, headache, vomiting, mutation, and carcinogenesis. Phenol-degrading bacterium DWB-1-8 was isolated from the activated sludge of textile wastewater; this strain was identified as Comamonas testosteroni by 16S rRNA gene sequencing. The optimal culture conditions for the cell growth and degradation of phenol were 0.7% K2HPO4, 0.6% NaH2PO4, 0.1% NH4NO3, 0.015% MgSO4·7H2O, 0.001% FeSO4·7H2O, an initial pH of 7, and a temperature of 30℃. The strain was also able to grow by using other toxic compounds, such as benzene, toluene, or xylene (BTX), as the sole source of carbon.

Rice Blast Control and Race Diversity by Mixed-Planting of Two Cultivars ('Hopyeongbyeo'/'Nampyeongbyeo') with Different Susceptibility to Magnaporthe oryzae (호평벼와 남평벼의 혼합재배에 의한 도열병 방제와 레이스 다양성의 변화)

  • Oh, In-Seok;Min, Ji-Young;Cho, Myung-Gil;Roh, Jae-Hwan;Shin, Dong-Bum;Song, Jin;Kim, Myeong-Ki;Cho, Young-Chan;Kim, Byung-Ryun;Han, Seong-Sook
    • Research in Plant Disease
    • /
    • v.14 no.3
    • /
    • pp.143-152
    • /
    • 2008
  • Mixed-planting of two rice cultivars, HP ('Hopyeongbyeo') and NP ('Nampyeongbyeo'), having a dissimilar susceptibility to rice blast was practiced for chemical-free control of rice blast in the field. The HP/NP combination was selected for applying under mechanized agricultural conditions. Because they have similar genetic characteristics such as seed germination and heading time, culm length, rice quality and size of rice grains except susceptibility to blast. Incidence of panicle blast was reduced 50.4 % compare with supposed blast incidence by HP/NP mixed-planting when the seeds of two cultivars were combined 1 to 1 as weight. Supposed blast incidence was estimated from reduction of rice blast caused by addition of a resistant cultivar NP. Race diversity of Magnaporthe oryzae was examined for correlation with control effect of HP/NP mixed-planting on rice blast. The population of dominant race KJ-101 was diminished and replaced with various co-existing races and eleven new races were appeared in mixed-planting plot. Total number of race isolated from mixed-planting plot was not largely different from mono-culture. However, detection frequency of the new race was increased and variation of the population size of each race was decreased in mixed-planting plots. It was shown that a biased community with a dominant race (KJ-101 or KI-181) was altered to a balanced one of coexisting races. From these results, it was supposed that the balanced diversity among co-existing races within a community might be correlated to control effect by HP/NP mixed-planting on rice blast. Further more, it should be studied that genetic characteristics of the individual race including a virulence on cv. HP and NP was examined for verifying a correlation of mixed-planting effect and race diversity.

Estimation for Red Pepper(Capsicum annum L.) Biomass by Reflectance Indices with Ground-Based Remote Sensor (지상부 원격탐사 센서의 반사율지수에 의한 고추 생체량 추정)

  • Kim, Hyun-Gu;Kang, Seong-Soo;Hong, Soon-Dal
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.42 no.2
    • /
    • pp.79-87
    • /
    • 2009
  • Pot experiments using sand culture were conducted in 2004 under greenhouse conditions to evaluate the effect of nitrogen deficiency on red pepper biomass. Nitrogen stress was imposed by implementing 6 levels (40% to 140%) of N in Hoagland's nutrient solution for red pepper. Canopy reflectance measurements were made with hand held spectral sensors including $GreenSeeker^{TM}$, $Crop\;Circle^{TM}$, and $Field\;Scout^{TM}$ Chlorophyll meter, and a spectroradiometer as well as Minolta SPAD-502 chlorophyll meter. Canopy reflectance and dry weight of red pepper were measured at five growth stages, the 30th, 40th, 50th, 80th and 120th day after planting(DAT). Dry weight of red pepper affected by nitrogen stress showed large differences between maximum and minimum values at the 120th DAT ranged from 48.2 to $196.6g\;plant^{-1}$, respectively. Several reflectance indices obtained from $GreenSeeker^{TM}$, $Crop\;Circle^{TM}$ and Spectroradiometer including chlorophyll readings were compared for evaluation of red pepper biomass. The reflectance indices such as rNDVI, aNDVI and gNDVI by the $Crop\;Circle^{TM}$ sensor showed the highest correlation coefficient with dry weight of red pepper at the 40th, 50th, and 80th DAT, respectively. Also these reflectance indices at the same growth station was closely correlated with dry weight, yield, and nitrogen uptake of red pepper at the 120th DAT, especially showing the best correlation coefficient at the 80th DAT. From these result, the aNDVI at the 80th DAT can significantly explain for dry weight of red pepper at the 120th DAT as well as for application level of nitrogen fertilizer. Consequently ground remote sensing as a non-destructive real-time assessment of plant nitrogen status was thought to be a useful tool for in season nitrogen management for red pepper providing both spatial and temporal information.

Effects of Manipulation Conditions on Development of Nuclear Transplant Bovine Embryos Derived from In Vitro Matured Oocytes (미세조작조건이 소 핵이식배의 발달에 미치는 영향)

  • 최상용;노규진;공일근;송상현;조성근;박준규;이효종;박충생
    • Korean Journal of Animal Reproduction
    • /
    • v.21 no.3
    • /
    • pp.293-302
    • /
    • 1997
  • Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

  • PDF

Evaluation of Pea (Pisum sativum L.) Genetic Resources for Sprout and Analysis of Optimum Culture Condition for Pea Sprout (완두나물 재배에 적합한 유용자원 선발과 완두나물 적정 재배조건 설정)

  • Koo, Sung-Cheol;Kang, Beom-Kyu;Kim, Hyun-Tae;Seo, Jeong-Hyun;Jeong, Kwang-Ho;Yun, Hong-Tae;Oh, In-Seok;Choi, Man-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.60 no.4
    • /
    • pp.484-490
    • /
    • 2015
  • Thirty-eight Pea (Pisum sativum L.) genotypes were screened to identify varieties to be suitable for sprout. Based on seed yield and sprout qualities such as whole length and sprout yield, five genotypes (PI269803, PI343278, PI343283, PI343300 and PI 343307) were primarily selected as candidates for pea sprouts. In order to determine optimal cultivation condition for pea sprouting, growth characteristics were investigated according to the change of germination temperature and days for sprouting. Whole length and hypocotyl length were observed to increase as a time dependent manner at each tested temperature (20, 23, and $25^{\circ}C$). However, whole length, hypocotyl length, and sprout yield were highly increased at $23^{\circ}C$ compared to 20 and $25^{\circ}C$. Especially, PI269803 and PI343300 showed higher sprout yield than the others. In addition, the effect of the change of germination temperature on antioxidant properties was estimated by measuring total phenolic content (TPC) and free radical scavenging activity (DPPH and ABST activity). TPC and DPPH/ABST activities of PI269803 and PI343300 were higher at $23^{\circ}C$ than at 20 and $25^{\circ}C$, while antioxidant properties of PI343278 and PI343283 were decreased in a temperature-dependent manner. The results show a high degree of correlation between TPC and antioxidant activities and suggest that the temperature change for pea sprouting could be responsible for antioxidant properties. Taken together, these results provide optimal cultivation conditions for pea sprouting and suggest that PI269803 and PI343300 with high sprout yield and antioxidant properties could be used for pea sprouts.