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Transformation of Adult Mesenchymal Stem Cells into Cardiomyocytes with 5-azacytidine: Isolated from the Adipose Tissues of Rat (성체 백서의 지방조직에서 추출한 중간엽 줄기세포의 5-azacytidine을 이용한 심근세포 분화 유도)

  • Choe Ju-Won;Kim Yong-In;Oh Tae-Yun;Cho Dai-Yoon;Sohn Dong-Suep;Lee Tae-Jin
    • Journal of Chest Surgery
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    • v.39 no.7 s.264
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    • pp.511-519
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    • 2006
  • Background: Loss of cardiomyocytes in the myocardial infarction leads to regional contractile dysfunction, and necrotized cardiomyocytes in infracted ventricular tissues are progressively replaced by fibroblasts forming scar tissue. Although cardiomyoplasty, or implantation of ventricular assist device or artificial heart was tried in refractory heart failure, the cardiac transplantation was the only therapeutic modality because these other therapeutic strategies were not permanent. Cell transplantation is tried instead of cardiac transplantation, especially bone marrow is the most popular donated organ. But because bone marrow aspiration procedure is invasive and painful, and it had the fewer amounts of cellular population, the adipose tissue is recommended for harvesting of mesenchymal stem cells. Material and Method: After adipose tissues were extracted from abdominal subcutaneous adipose tissue and intra-abdominal adipose tissue individually, the cellular components were obtained by same method. These cellular components were tried to transformation with the various titers of 5-azacytidine to descript the appropriate concentration of 5-azacytidine and possibility of transformation ability of adipose tissue. Group 1 is abdominal subcutaneous adipose tissue and Group 2 is intra-abdominal adipose tissue-retroperitoneal adipose tissue and omentum. Cellular components were extracted by collagenase and $NH_4Cl$ et al, and these components were cultured by non-induction media - DMEM media containing 10% FBS and inducted by none, $3{\mu}mol/L,\;6{\mu}mol/L,\;and\;9{\mu}mol/L$ 5-azacytidine after the 1st and 2nd subculture. After 4 weeks incubation, tile cell blocks were made, immunostaining was done with the antibodies of CD34, heavy myosin chain, troponin T, and SMA. Result: Immunostaining of the transformed cells for troponin T was positive in the $6{\mu}mol/L\;&\;9{\mu}mol/L$ 5-azacytidine of Group 1 & 2, but CD34 and heavy myosin chain antibodies were negative and SMA antibody was positive in the $3{\mu}mol/L\;&\;6{\mu}mol/L$ 5-azacytidne of Group 2. Conclusion: These observations confirm that adult mesenchymal stem cells isolated from the abdominal subcutaneous adipose tissues and intra-abdominal adipose tissues can be chemically transformed into cardiomyocytes. This can potentially be a source of autologous cells for myocardial repair.

Effects of Activation Treatments and Culture Condition on In Vitro Development of Caprine In Vivo and In Vitro Oocytes (재래산양의 체내 및 체외유래 난자의 활성화 처리방법 및 배양조건이 단위발생란의 체외발달에 미치는 영향)

  • Park H. S.;Kim T. S.;Lee Y. H.;Jung S. Y.;Lee M. Y.;Jin J. I.;Park J. K.;Lee J. S.;Kim C. H.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.181-185
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    • 2004
  • This study was conducted to examine whether activation treatments, source of oocytes and culture conditions affect in vitro developmental ability of caprine oocytes. Mature Korean native goats were pretreated with intravaginal CIDR for 10 days. The goats were then treated with a single intramuscular injection of 1,000 IU PMSG on Day 8 or twice daily injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR insertion for superovulation. All the goats were injected with 10 mg PGF/sub 2a/ on Day 8 and 400 IU hCG on Day 10 of CIDR. Oocytes were surgically collected by oviduct flushing(in vivo maturation) or direct follicle aspiration(in vitro maturation) through mid-ventral incision at 35 h after hCG injection. Fifteen to twenty oocytes were placed in TCM-199 medium containing 25 mM Hepes and hormones under mineral oil at 39℃ in a humudified atmosphere of 5% CO₂ in air for 22 to 24 h. After maturation, the oocytes were activated by electric stimulation or ionomycin + 6-DMAP. The activated oocytes were then cultured in M16, TCM-199 and mSOF media supplemented with proteins at 39℃ for 6 to 7 days. Activation treatments did not affect cleavage of the oocytes. The cleavage rates were 64.1% (41/64) in oocytes activated by electric stimulation and 76.5% (218/285) in oocytes activated by ionomycin + 6-DMAP. The proportion of development to blastocyst was 15.6% (34/218) in oocytes activated by ionomycin + 6-DMAP, but activation by electric stimulation did not support embryos developed beyond morula stage. There were no differences in the cleavage rates of activated oocytes experiencing in vivo (86.8%, 66/76) and in vitro maturation (69.0%, 127/184). However, the development rate to blastocyst stage was significantly (P<0.05) higher for oocytes matured in vivo (50.0%, 33/66) compared to in vitro (0.8%, 1/127). Culture conditions did not affect the cleavage of -activated oocytes. The cleavage rates were 51.6% (49/95) in M16, 64.3% (18/28) in TCM-199 and 81.0% (145/179) in mSOF, respectively. By contrast, the development rate of activated oocytes to stage was greater (P<0.05) for oocytes cultured in mSOF medium (23.4%, 34/145) than in M16 or TCM-199 (0.0%). Our results suggest that source of oocytes and culture conditions are major factors affecting in vitro development of caprine parthenogenetic oocytes.

Effects of Dissolved Compounds in Groundwater on TCE Degradations Reaction by Nanoscale Zero-Valent Iron (나노영가철의 TCE 분해반응 시 지하수 용존물질의 영향)

  • Kim, Tae-Ho;Kim, Hong-Seok;Lee, Jin-Yong;Cheon, Jeong-Yong;Lee, Kang-Kun;Hwang, In-Seong
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.6
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    • pp.413-419
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    • 2011
  • Nanoscale zero-valent iron (NZVI) particles were tested as remediation media for groundwater contaminated by organic pollutants (e.g., TCE, trichloroethylene). The contaminated groundwater contained anions ($NO_3^-$, $Cl^-$, $SO_4^{2-}$, and $HCO_3^-$) and natural organic matter (NOM). Treatability of commercial NZVI particles (NANOFER 25, Nanoiron, Czech) was tested by using a synthetic groundwater and the field groundwater samples. More than 95% of 1.8 mM TCE was removed within 20 hours with a NZVI dosage of 25 g/L ($k=0.15hr^{-1}$). Repetitive degradation experiments revealed that the removal capacity of NANOFER 25 was 0.19 mmole TCE/g NZVI. TCE degradation reactions were not substantially affected by the presence of each anion with concentrations as high as 100 times the average field concentrations. However, when the four anions ($NO_3^-$, $Cl^-$, $SO_4^{2-}$, $HCO_3^-$) were present simultaneously. the degradation reactivity and removal capacity were decreased by 60% ($k=0.069hr^{-1}$) and 10%, respectively. The k value of TCE degradation in the presence of NZVI (25 g/L) with dissovled organic carbon of 2.5 mg/L was also decreased by 84% ($k=0.025hr^{-1}$). In the experiments with the field groundwater, more than 90% of $1.8{\mu}M$ TCE, which is the concentration of TCE at the source zone, was removed within 10 hours with a NANOFER 25 dosage of 25 g/L. The results imply that the contaminated groundwater can effectively be treated by NANOFER 25 with more information on the hydrogeology of the site.

Characteristics of Culture of Oyster Mushroom (Pleurotus ostreatus) on Addition Rate of Acer tegmentosum (산겨릅나무(Acer tegmentosum) 톱밥의 첨가량에 따른 느타리버섯의 재배적 특성)

  • Lee, Chan-Jung;Jhune, Chang-Sung;Cheong, Jong-Chun;Moon, Ji-Won;Kong, Won-Sik;Suh, Jang-Sun;Park, Gi-Chun;Shin, Yu-Su
    • The Korean Journal of Mycology
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    • v.41 no.1
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    • pp.21-27
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    • 2013
  • This study was carried out to investigated optimum mixing ratio of Korean natural Acer tegmentosum for production of functional oyster mushroom. Total nitrogen and carbon sources of Acer tegmentosum were 0.2% and 44.4%, respectively and C/N ratio was 234. Total nitrogen source and pH of substrate mixed with Acer tegmentosum were 2.7~2.9 and 4.8~5.0, respectively. The contents of $P_2O_5$, $K_2O$ and MgO at Acer tegmentosum media were higher than that of the control. Mycerial growth was the fastest at Acer tegmentosum 10%, and slower by increase of Acer tegmentosum substrate. Yields of fruiting body was the highest to 159 g/850 mL at 10% of Acer tegmentosum and dimeter and thick of pileus were the highest, too. The L value of pileus and stipes were decreased by increase of Acer tegmentosum substrate, but there was no significant difference in the a-value and the b-value. The contents of $P_2O_5$ and $K_2O$ of fruiting body were increased at Acer tegmentosum substrate, but there was no significant difference in contents of CaO, MgO and $Na_2O$.

Dose Verification Study of Brachytherapy Plans Using Monte Carlo Methods and CT Images (CT 영상 및 몬테칼로 계산에 기반한 근접 방사선치료계획의 선량분포 평가 방법 연구)

  • Cheong, Kwang-Ho;Lee, Me-Yeon;Kang, Sei-Kwon;Bae, Hoon-Sik;Park, So-Ah;Kim, Kyoung-Joo;Hwang, Tae-Jin;Oh, Do-Hoon
    • Progress in Medical Physics
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    • v.21 no.3
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    • pp.253-260
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    • 2010
  • Most brachytherapy treatment planning systems employ a dosimetry formalism based on the AAPM TG-43 report which does not appropriately consider tissue heterogeneity. In this study we aimed to set up a simple Monte Carlo-based intracavitary high-dose-rate brachytherapy (IC-HDRB) plan verification platform, focusing particularly on the robustness of the direct Monte Carlo dose calculation using material and density information derived from CT images. CT images of slab phantoms and a uterine cervical cancer patient were used for brachytherapy plans based on the Plato (Nucletron, Netherlands) brachytherapy planning system. Monte Carlo simulations were implemented using the parameters from the Plato system and compared with the EBT film dosimetry and conventional dose computations. EGSnrc based DOSXYZnrc code was used for Monte Carlo simulations. Each $^{192}Ir$ source of the afterloader was approximately modeled as a parallel-piped shape inside the converted CT data set whose voxel size was $2{\times}2{\times}2\;mm^3$. Bracytherapy dose calculations based on the TG-43 showed good agreement with the Monte Carlo results in a homogeneous media whose density was close to water, but there were significant errors in high-density materials. For a patient case, A and B point dose differences were less than 3%, while the mean dose discrepancy was as much as 5%. Conventional dose computation methods might underdose the targets by not accounting for the effects of high-density materials. The proposed platform was shown to be feasible and to have good dose calculation accuracy. One should be careful when confirming the plan using a conventional brachytherapy dose computation method, and moreover, an independent dose verification system as developed in this study might be helpful.

Effect of Dietary Supplementation of Fresh Water Algae Euglena on the Performance and Egg Quality and Fatty Acid Composition of Egg Yolk in Laying Hens (산란계에서 담수녹조류 Euglena의 첨가사료가 생산성 및 계란의 품질과 지방산 조성에 미치는 영향)

  • Choi S. W.;Park I. K.;Park B. S.
    • Korean Journal of Poultry Science
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    • v.31 no.4
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    • pp.283-291
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    • 2004
  • Feeding trials were conducted with Euglena strains grown under different media. The effect of supplementation of Euglena on the laying performance, egg quality and fatty acid composition of egg yolk was studied. In experiment I, two hundred eighty 32-wk-old ISA Brown layers were randomly assigned to seven dietary treatments for 4 wks. Each treatment consisted of 4 replications with 10 birds each housed in two birds cages. Control diet was formulated to have $17\%$ CP and 2,750 kcal ME/kg. Euglena gracilis Z. (EG) was added to control diet at the level of 0.25, 0.5, $1.0\%$ and Euglena gracilis Z. bleached and DHA enriched (EGBD; a strain mutated by streptomycin and cultivated in DHA enriched medium) at the level of 0.5, 1.0, $2.0\%$ in the diet. In experiment 2, three hundred 84-wk-old ISA brown layers were randomly assigned to five dietary treatments: T1; Control, T2; T1 + EGBD $0.5\%$, T3; T1 + Euglena gracilis Z. DHA enriched (EGD; cultivated in DHA enriched medium) $0.5\%$, T4; T1 + EGD $1.0\%$, T5; T1 + EGD $2.0\%$. Each treatment had 5 replication of 12 birds each housed in two birds cages. In experiments 1 and 2, Euglena suppplementation did not significantly affect egg production but increased egg weight and feed intake. In experiment 1, EG was more effective in increasing egg yolk color score than EGBD. Egg yolk color of EG $1\%$ treatment showed the highest score. EGBD supplementation increased DHA concentration of egg yolk. EGBD $2\%$ treatment showed the highest DHA and the lowest palmitic and stearic acids concentration in the egg yolk. In experiment 2, EGBD $0.5\%$ treatment showed highest DHA level in egg yolk (P<0.05). It was conducted that EGBD is a single cell protein source rich in DHA, that can be used to produce DHA enriched eggs.

독창적 아이디어에서 창조적 혁신까지 : 인공씨감자 기술혁신 성공사례 분석

  • 현재호
    • Proceedings of the Technology Innovation Conference
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    • 1997.07a
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    • pp.222-223
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    • 1997
  • By analyzing the successful innovation case of potato microtuber mass production technology, a representative case of technology-push type creative innovation in an imitation oriented research culture, this paper attempts to figure out conceptual model of creative innovation that is initiated by the public laboratories in catching-up country, Stages of creative innovation can be divided into the internal R&D stage and the external commercialization stage. Success of the internal R&D stage depended on autonomy to secure creative research idea and commitment of individual researchers. Psychological pressure evoked from sportlights of mass media and commitment of sponsor increased the intensity of research efforts of the researcher Recognition of research problem and its significance was intensified by site visits of agricultural fields, and the recognized higher impacts of expected research results and knowledge creation achieved were a fundamental source of self-motivation. In the stage of commercialization stage, various legal, socio-economic, and psychological barriers were confronted. In a catching-up country lacking of experiences of creative innovation, creative innovation process can be regarded as a barrier elimination and cultural revolution process. Among the barriers, psychological refusal of farmers to corn-sized potato seeds was critical, which finally enforced to further researches to enlarge the size of potato seeds. In addition, the researcher has concentrated his research efforts in one specialized research area by getting a series of similar research project funds rather than diversification. It was lucky for him to have a chance to carry out a series of similar researches in one research area during the last 10 years. In getting research funds from government and private companies continuously in one research area, both internal and external promoters played significant roles.

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Carbohydrate Metabolism in Preimplantation Stage Embryos and the Role of Metabolites (착상전 초기 배아에서 탄수화물 대사와 그 대사물의 역할)

  • Cheon, Yong-Pil
    • Development and Reproduction
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    • v.12 no.1
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    • pp.19-30
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    • 2008
  • Proper development of fertilized oocyte to blastocyst is a key step in mammalian development to implantation. During development of preimplantation embryos, the mammalian embryo needs supply the energy substrate for keep viability. Usually mammalian oocyte get substrate especially energy substrate from oviduct and uterus, because it does not store much substrate into cytoplasm during oogenesis. Carbohydrates are known as a main energy substrate for preimplantation stage embryos. Glucose, lactate and pyruvate are essential component in preimplantation embryo culture media and there are stage specific preferences to them. Glucose transporter and $H^+$-monocarboxylate cotransporter are a main mediator for carbohydrate transport and those expression levels are primarily under the control of intrinsic or extrinsic factors like insulin and glucose. Other organic substances, amino acids, lipids and nucleotides are used as energy substance and cellular regulation factor. Though since 1960s, successful development of fertilized embryo to blastocyst has been accomplished with chemically defined medium for example BWW and give rise to normal offspring in mammals, the role of metabolites and the regulation of intermediary metabolism are still poorly understood. Glucose may permit expression of metabolic enzymes and transporters in compacting morula, capable of generating the energy required for blastocyst formation. In addition, it has been suggested that the cytokines can modulate the metabolic rate of carbohydrate in embryos and regulate the preimplantation embryonic development through control the metabolic rate. Recently we showed that lactate can be used as a mediator for preimplantation embryonic development. Those observations indicate that metabolites of carbohydrate are required by the early embryo, not only as an energy source, but also as a key substrate for other regulatory and biosynthetic pathways. In addition metabolites of carbohydrate may involve in cellular activity during development of preimplantation embryos. It is suggested that through these regulation and with other regulation mechanisms, embryo and uterus can prepare the embryo implantation and further development, properly.

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The Study on A Peculiarity of Mise-en-scene Found in Animation :Focused on Russian Animation (애니메이션 미장센 특성 연구 - 러시아 애니메이션을 중심으로)

  • Kim, MiRNaRae;Min, JunIl
    • Cartoon and Animation Studies
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    • s.44
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    • pp.1-31
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    • 2016
  • In this thesis, the movie with mise-en-scene established was compared with the peculiarity of the play that is the etymological source of the term to identify the peculiarity of mise-en-scene which was substituted into animation to find the peculiarity of mise-en-scene in animation. To emphasize the direct connection between the frame's visual peculiarity and the director's opinions, the mise-en-scene of director centered animation created under a restricted environment was reviewed. Mise-en-scene which started from movie critics theory does not simply mean the arrangement of images in a frame. Mise-en-scene emphasizes the exposure of the work's motive by the visual components. The animation's assuming the middle point of environmental share possessed by play and movie when schematizing the genre peculiarity of animation, play and movie was a noteworthy result. It can be said that the cause is that the animation's peculiarity yield different results depending on the making methods; we verified that this is a key factor in the analysis of animation's mise-en-scene. I emphasized that the peculiarity of animation mise-en-scene is in its making method and material and suggested identifying the work's making methods and analyzing the work's aesthetic results derived in this way. The russian animation which was perceived as peripheral arts was relatively free from the burden of censorship while receiving support from the Soviet as a media for propaganda. The russian animation's mise-en-scene which found the material for its works in the country's folklore was metaphorical, focused on new expression forms and achieved experimental elements. Russian animation pursues a unique aesthetic world through space expression based on the forms of opera or ballet and heavy motions formed static inbetweens.

Biodegradation of Aromatic Compounds by Strains of Pseudomonas (Pseudomonas속 세균에 의한 방향족화합물 생분해)

  • 정윤창;김경남;최용진;양한철;송준상;서윤수
    • Microbiology and Biotechnology Letters
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    • v.17 no.2
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    • pp.100-108
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    • 1989
  • Thirty-six aromatic compound biodegraders; 10 strains for benzoate, 10 for salicylate, 6 for m-toluate, and 10 for DL-camphor were isolated and taxonomically characterized. A mutant Pseudomonas strain, Ben 6-2, derived from Ben 6 revealed remarkably improved ability to metabolize benzoate. Thus enhancement of the average substrate removal rate from 5.2 to 11.0mg/$\ell$/ hr was attained by the mutant. Both of strains Sal 7 and Tol 2, degraders of salicylate and m-toluate respectively, were classified as Pseudomonas sup. Both strains were found to be extremely effective in metabolizing each aromatic substrates. The average substrate degradation rates in minimal salt media containing 2,200mg/$\ell$ of the substrate were calculated to be 40.1 mg/$\ell$/ hr for strain Sal 7 and 33.0mg/$\ell$/ hr for Tol 2. Cam 10, a camphor degrading strain was demonstrated to be capable of mineralizing benzoate, phenol, toluene, octane, cyclohexane and xylene as well as camphor. Strain 1040 isolated from Cam 10 after repented adaptation to 1,000 mg/$\ell$ m-toluate gained the ability to utilize toluate as a sole carbon source. The mutant Brew actively at the expense of a mixture of car-bon sources; camphor, m-toluate, benzoate and phenol (each: 200 mg/$\ell$) and utilized the substances in the preferential order of camphor, phenol, benzoate, and m-toluate. Among the biodegraders examined Cam 1040 and Tol 2 were detected to harbor plasmid. The plasmid from Cam 1001 was determined to be about 98kb, and evidenced to encode the enzyme(s) for the degradation of camphor. For the further diversification of the metabolic potentials of Cam 1040, the NAH 2 plasmid of Pseudomonas putida NCIB 9816 was transferred to Cam 1040 by conjugation. The exconjugant obtained, Cam 1043, proved to gain an additional ability to metabolize salicylate and naphthalene.

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