• Title/Summary/Keyword: seeding density

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A Novel Bioassay System for Screening of Compounds Affecting Anthocyanin Biosynthesis Pathway in White Corn Leaf Segment (백화옥수수 잎절편을 이용한 안토시아닌 생합성 조절제 탐색용 신규 검정법 확립)

  • Kim, Jin-Seog;Lee, Byung-Hoi;Lee, Jung-Ae;Oh, Kwang-Hoon;Cho, Kwang-Yun
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.207-214
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    • 2003
  • This study was carried out to establish a novel bioassay system for screening of compounds affecting aromatic amino acid or anthocyanin biosynthesis through investigating a degree of sucrose-induced anthocyanin formation such as size of plant material, buffer conditions, light intensity and irradiated duration, incubation temp., etc were determined and standard procedure (suitable experimental condition) was set up as follows. The second leaf blade of white corn seeding induced by fluridone treatment were segmented into a size of 5${\times}$5 min. The segments were floated on the solution of 1% sucrose in 1.0mM MES buffer (pH6.0∼6.5) and incubated at 26$^{\circ}C$ for 2days under the continuous light condition(70∼100$\mu$mol m$^{-2}$ s$^{-1}$ ). Anthocyanin in the purpled tissues was extracted with methanol containing 1% HCl and the optical density of the clear supematants was determined at 528mm. Influences of some chemicals were tested using this system. Glyphosate, 5-enolpyruvylshikimate 3-phosphate synthase inhibitor, showed most sensitive response with I$_{50}$ value at 3.3$\mu$M. Dicyclohexylcarbodiimide(DCCD) and parachloromercuribenzenesulfonic acid(PCMBS) had a relatively strong ingibition with I50 value at 7.1$\mu$M and 10.2$\mu$M, respectively. These results show that sucrose-induced anthocyanin formation in white com leaf segment provide a very simple and rapid system for searching new compounds affecting aromatic amino acid or anteocyanin biosynthesis by screening at less than 10$\mu$M.

Effect of Crop Establishment and Weed Control Method on Productivity of Transplanted aman Rice

  • Ali, Mohammad;Haque Bir, Md. Shahidul;Rahman, Md. Habibur;Ayesha, Sultana Kaniz;Hoque, Aminul;Harun-Ar-Rashid, Md.;Islam, Md. Rashidul;Park, Kee Woong
    • Weed & Turfgrass Science
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    • v.7 no.2
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    • pp.120-129
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    • 2018
  • This experiment was conducted to find the most suitable crop establishment method and weed management practices for transplanted aman rice in Bangladesh. Rice variety Bangladesh Rice Research Institute (BRRI) dhan44 was used as planting materials where three crop establishment methods ($T_1$: direct wet seeding by drum seeder; $T_2$: hand broadcasting; $T_3$: transplanting) and four weeding options ($W_1$: Hand weeding (HW); $W_2$: BRRI weeder+HW; $W_3$: Herbicide+HW; and $W_4$: no weeding) were tested. Among the crop establishment methods, the highest grain yield ($5.12t\;ha^{-1}$) was obtained with the $T_3$, while the highest benefit cost ratio (BCR) of 2.08 was found in $T_2$. In case of the weed management method, $W_1$ showed superior results on the plant $density/m^2$ (139.66) at 60 days after transplanting (DAT), grain yield ($4.97t\;ha^{-1}$), and BCR (2.03). On the other hand, the highest plant dry matter (36.20 g) at 60 DAT and the highest yield ($6.10t\;ha^{-1}$) were obtained in a $T_3W_1$ combination. The results of this study show that the productivity of rice during aman season could be most significantly increased with the use of transplanting ($T_3$) alone, hand weeding ($W_1$) alone, or a combination of the two methods ($T_3W_1$).

Artificial seed production and cultivation of Sargassum macrocarpum (Fucales, Phaeophyta)

  • Ko, Shin Ja;Kim, Yoo Kyung;Hong, Seong Wan;Kang, Min Su;Park, Chan Sun;Hwang, Eun Kyoung;Lee, Young Don
    • ALGAE
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    • v.35 no.2
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    • pp.123-131
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    • 2020
  • Sargassum macrocarpum is a rich source of anti-inflammatory compounds. Recently, one of the compounds, tuberatolide B, has been reported as a functional anti-inflammatory additive for foods and nutraceuticals. The artificial seeding, growth and maturation of S. macrocarpum were investigated from May 2018 to September 2019. Indoor culture experiments for induction of egg release were conducted at temperatures of 17, 20, 23, and 26℃ and irradiances of 0, 10, 20, 40, and 80 μmol photons m-2 s-1 under 14 : 10 h (L : D) photoperiod. Within a given treatment combination, higher temperatures and irradiance levels favoured the maturation of receptacles in S. macrocarpum. Using artificial temperature and irradiance control, thalli matured one month earlier than thalli in nature. Under natural condition, receptacle formation began in April, and the eggs were released in June and July. The release of eggs from the receptacles was promoted at 17-20℃ and 40-80 μmol photons m-2 s-1, and the fastest growth of germlings occuring at 15-17℃ and 40 μmol photons m-2 s-1. For mature thalli, 300 g wet-weight was sufficient to seed 100 m of seed string. Thalli grew to 10.5 ± 2.6 cm in length at a density of 6.7 ± 3.3 individuals m-1 after 1 year of cultivation, from germination. This study demonstrates that it is possible to cultivate S. macrocarpum for the production of anti-inflammatory products.

Enhanced Chondrogenic Differentiation of Human Adipose-derived Stem Cells with Inverse Opal Scaffolds (역오팔 구조 지지체를 이용한 인간 지방 유래 줄기 세포의 연골 분화 촉진)

  • Bhang, Suk Ho;Yu, Taekyung
    • Korean Chemical Engineering Research
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    • v.51 no.6
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    • pp.727-732
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    • 2013
  • In this report, we present an inverse opal scaffold that can enhance the chondrogenic differentiation of human adipose-derived stem cells (hADSCs) without drug, gene, or cytokine supplement. Inverse opal scaffolds based on poly(D,L-lactide-co-glycolide) were formed with uniform $200{\mu}m$ pores. Due to uniform pore sizes and well-controlled interconnectivity of inverse opal scaffold, hADSCs were allowed to distribute homogeneously throughout the scaffolds. As a result, high cell density culture with scaffold was possible. Since the hADSCs cultured in inverse opal scaffolds were subjected to limited supplies of oxygen and nutrients, these cells were naturally preconditioned to a hypoxic environment that stimulated the up-regulation of hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$). As a result, apoptotic activity of hADSCs until 3 weeks after initial cell seeding was significantly reduced and chondrogenic differentiation related molecular signal cascades were up regulated (transforming growth factor-beta, phosphorylated AKT, and phosphorylated p38 expression). In contrast, hADSCs cultured with small and non-uniform porous scaffolds showed significantly increased apoptotic activity with decreased chondrogenic differentiation. Taken together, inverse opal scaffold could potentially be used as an effective tool for improving chondrogenesis using stem cells.

Characteristics of Power Generation and Organic Matter Removal in Air-Cathode MFC with respect to Microbial Concentration (미생물 농도에 따르는 Air-Cathode MFC의 전력발생과 유기물질제거 특성)

  • Kim, Doyoung;Lim, Bongsu;Choi, Chansoo;Kim, Daehyun
    • Journal of Korean Society on Water Environment
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    • v.28 no.6
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    • pp.917-922
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    • 2012
  • In order to improve applicability of a microbial fuel cell the laboratory-scaled study has been performed by adopting an air-cathode MFC system with high concentrated anaerobic slugies in this study. The concentrations of microbes are grouped into three types, Type A (TS 1.7%), Type B (TS 1.1%) and Type C (TS 0.51%). The open circuit voltage $(V_{oc})$ characteristics showed that the medium microbes concentration of 1.10% (Type B) kept a constant voltage of 1.0 V for 150 hours, which showed the longest time among three types (Type A and Type C). The discharge charge curves for a closed circuit with $500 \Omega$ also showed that Type B generated a stable discharge voltage of 0.8 V for a longer time as in the open circuit voltage case. This could be explained by the relatively large amount of the attached microbes. Under the $V_{oc}$condition the COD removal efficiency of Type B was found to be low for a long time, but those of Type A and C were found to be high for a short period of time. Therefore, the suspended microbes could decrease the coulombic efficiency. It was concluded that the high $V_{oc}$ was caused by low COD and the $V_{oc}$ became low after the COD removal. The COD reduction resulted in an unstable and low working voltage. From the polarization characteristics Type A was found to show the highest power density of $193\;mW/m^2$ with a fill factor of 0.127 due to the relatively high remaining COD even after the MFC reaction.

Identification of Niche Conditions Supporting Short-term Culture of Spermatogonial Stem Cells Derived from Porcine Neonatal Testis

  • Park, Min Hee;Park, Ji Eun;Kim, Min Seong;Lee, Kwon Young;Yun, Jung Im;Choi, Jung Hoon;Lee, Eunsong;Lee, Seung Tae
    • Journal of Embryo Transfer
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    • v.29 no.3
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    • pp.221-228
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    • 2014
  • Despite that porcine spermatogonial stem cells (pSSCs) have been regarded as a practical tool for preserving eternally genetic backgrounds derived from pigs with high performance in the economic traits or phenotypes of specific human diseases, there were no reports about precise definition of niche conditions promoting proliferation and maintenance of pSSCs. Accordingly, we tried to determine niche conditions supporting proliferation and maintenance of undifferentiated pSSCs for short-term. For these, undifferentiated pSSCs were progressively cultured in different composition of culture medium, seeding density of pSSCs, type of feeder cells and concentration of growth factors, and then total number of and alkaline phosphatase (AP) activity of pSSCs were investigated at post-6 day culture. As the results, the culture of $4{\times}10^5$ pSSCs on mitotically in activated $2{\times}10^5$ STO cells in the mouse embryonic stem cell culture medium (mESCCM) supplemented with 30 ng/ml glial cell line-derived neurotrophic factor (GDNF) was identified as the best niche condition supporting effectively the short-term maintenance of undifferentiated pSSCs. Moreover, the optimized short-term culture system will be a basis for developing long-term culture system of pSSCs in the following researches.

Tumorsphere formation and cancer stem cell characterization of REM134 canine mammary carcinoma cells (개 REM134 유선종양세포의 sphere 형성을 통한 암 줄기세포 특성 분석)

  • Byeon, Jeong Su;Lee, Jienny;Jeong, Da-Un;Gu, Na-Yeon;Cho, In-Soo;Cha, Sang-Ho
    • Korean Journal of Veterinary Research
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    • v.58 no.4
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    • pp.201-209
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    • 2018
  • Canine mammary tumors are among the most frequently observed cutaneous tumors in female dogs. Cancer stem cells (CSCs), referred to as tumor-initiating cells, are thought to have properties similar to normal stem cells such as the ability to self-renewal and to differentiate into various cell types. Biological understanding of CSCs and the critical pathways involved in their maintenance are important in research and therapy for mammary tumors. We conducted the present study on sphere formation from REM134 cells by using methylcellulose to produce tumorspheres on a large scale and compared the specific markers of the spheres-formed and plating-cultured REM134 cells. The results revealed that the tumorspheres cultured in methylcellulose had higher seeding density and improved morphology compared to those produced in normal sphere formation medium. Expression levels of stemness markers and CSC-related markers were higher in tumorsphere-forming cells than in plating-cultured cells. Subsequently, we transplanted the tumorsphere-forming and plating-cultured cells into female nude mice to examine their tumorigenic potential. Tumor volume increased rapidly in mice transplanted with tumorsphere-derived cells compared to plating-cultured cells. We observed a novel sphere-forming condition for REM134 cells and showed that REM134 cell tumorspheres can exhibit improved CSC properties.

Tight junctional inhibition of entry of Toxoplasma gondii into MDCK cells (MDCK세포의 tight junction 형성이 Toxoplusmu gondii의 숙주세포 침투에 미치는 효과)

  • 남호우;윤지혜
    • Parasites, Hosts and Diseases
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    • v.28 no.4
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    • pp.197-206
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    • 1990
  • Various conditions of cultures were performed to investigate the role of tight junctions formed between adjacent MDCK cells on the entry of Toxoplasma. When MDCK cells were cocultured with excess number of Toxoplasma at the seeding density of 1×105, 3×105, and 5×105 cells/ml for 4 days, the number of intracellular parasites decreased rapidly as the host cells reached saturation density, i.e., the formation of tight junctions. When the concentration of calcium in the media (1.8 mM in general) was shifted to $5{\mu}M$ that resulted in the elimination of tight junction, the penetration of Toxoplasma increased about 2-fold(p<0.05) in the saturated culture, while that of non-saturated culture decreased by half. Trypsin-EDTA which was treated to conquer the tight junctions of saturated culture favored the entry of Toxoplasma about 2.5-fold(P<0.05) compared to the non-treated, while that of non- saturated culture decreased to about one fifth. It was suggested that the tight junctions of epithelial cells play a role as a barrier for the entry of Toxoplasma and Toxoplasma penetrate into hoot cells through membrane structure-specific, i.e., certain kind of receptors present on the basolateral rather than apical surface of MDCK cells.

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Fabrication of Gd1.5Ba2Cu3O7-y Bulk Superconductors from the Powder Synthesized by a Solid-State Reaction Method (고상반응법으로 합성한 분말로부터 Gd1.5Ba2Cu3O7-y 벌크 초전도체의 제조)

  • Kim, Yong Ju;Park, Seung Yeon;You, Byung Youn;Park, Soon-Dong;Kim, Chan-Joong
    • Korean Journal of Materials Research
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    • v.23 no.6
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    • pp.309-315
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    • 2013
  • $GdBa_2Cu_3O_{7-y}$(Gd123) powders were synthesized by the solid-state reaction method using $Gd_2O_3$ (99.9% purity), $BaCO_3$ (99.75%) and CuO (99.9%) powders. The synthesized Gd123 powder and the Gd123 powder with $Gd_2O_3$ addition ($Gd_{1.5}Ba_2Cu_3O_{7-y}$(Gd1.5)) were used as raw powders for the fabrication of Gd123 bulk superconductors. The Gd123 and Gd1.5 bulk superconductors were fabricated by sintering or a top-seeded melt growth (TSMG) process. The superconducting transition temperature ($T_{c,onset}$) of the sintered Gd123 was 93 K and the transition width was as large as 20 K. The $T_{c,onset}$ of the TSMG processed Gd123 was 82 K and the transition width was also as large as 12 K. The critical current density ($J_c$) at 77 K and 0 T of the sintered Gd123 and TSMG processed Gd123 were as low as a few hundreds A/$cm^2$. The addition of 0.25 mole $Gd_2O_3$ and 1 wt.% $CeO_2$ to Gd123 enhanced the $T_c$, $J_c$ and magnetic flux density (H) of the TSMG processed Gd123 sample owing to the formation of the superconducting phase with high flux pinning capability. The $T_c$ of the TSMG processed Gd1.5 was 92 K and the transition width was 1 K. The $J_cs$ at 77 K (0 T and 2 T) were $3.2{\times}10^4\;A/cm^2$ and $2.5{\times}10^4\;A/cm^2$, respectively. The H at 77 K of the TSMG-processed Gd1.5 was 1.96 kG, which is 54% of the applied magnetic field (3.45 kG).

The Effect of the Plasma Treatment on PLGA Scaffold for Adhesion and Chondrogenic Differentiation of Human Adipose-derived Stromal Cells (인체지방유래 간질세포의 부착 및 연골분화유도를 위한 PLGA 지지체의 플라즈마 처리 효과)

  • Dong, Chun Ji;Jun, Young Joon;Cho, Hyun Mi;Oh, Deuk Young;Han, Dong Keun;Rhie, Jong Won;Ahn, Sang Tae
    • Archives of Plastic Surgery
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    • v.33 no.1
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    • pp.46-52
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    • 2006
  • High-density micromass culture was needed to take three dimensions culture with ASCs(adipose derived stromal cells) and chondrogenesis. However, the synthetic polymer has hydrophobic character and low affinity to cells and other biomolecules. Therefore, the surface modification without changes of physical and chemical properties is necessary for more suitable condition to cells and biomolecules. This study was performed to investigate the effect of surface modification of poly (lactic-co-glycolic acid)(PLGA) scaffold by plasma treatment (P(+)) on the adhesion, proliferation and chondrogenesis of ASCs, and not plasma treatment (P(-)). ASCs were isolated from human subcutaneous adipose tissue obtained by lipectomy and liposuction. At 1 hour 30 minutes and 3days after cell seeding onto the P(-) group and the P(+) group, total DNA amount of attached and proliferated ASCs markedly increased in the P(+) group (p < 0.05). The changes of the actin under confocal microscope were done for evaluation of cellular affinity, at 1 hour 30 minutes, the shape of the cells was spherical form in all group. At 3rd day, the shape of the cells was fiber network form and finely arranged in P(+) group rather than in P(-) group. RT-PCR analysis of cartilage-specific type II collagen and link protein were expressed in 1, 2 weeks of induction. Amount of Glycoaminoglycan (GAG) markedly increased in P(+) group(p < 0.05). In a week, extracellular matrix was not observed in the Alcian blue and Safranin O staining. However in 2 weeks, it was observed that sulfated proteoglycan increased in P(+) group rather than in P(-) group. In conclusion, we recognized that plasma treatment of PLGA scaffold could increase the hydrophilic property of cells, and provide suitable environment for high-density micromass culture to chondrogenesis