• Title/Summary/Keyword: Cell formation

Search Result 4,024, Processing Time 0.037 seconds

Isolation of Bacteria Associated with the King Oyster Mushroom, Pleurotus eryngii

  • Lim, Yun-Jung;Ryu, Jae-San;Shi, Shanliang;Noh, Won;Kim, Eon-Mi;Le, Quy Yang;Lee, Hyun-Sook;Ro, Hyeon-Su
    • Mycobiology
    • /
    • v.36 no.1
    • /
    • pp.13-18
    • /
    • 2008
  • Eight distinct bacteria were isolated form diseased mycelia of the edible mushroom, Pleurotus eryngii. 16S rDNA sequence analysis showed that the isolates belonged to a variety of bacterial genera including Bacillus (LBS5), Enterobacter (LBS1), Sphingomonas (LBS8 and LBS10), Staphylococcus (LBS3, LBS4 and LBS9) and Moraxella (LBS6). Among them, 4 bacterial isolates including LBS1, LBS4, LBS5, and LBS9 evidenced growth inhibitory activity on the mushroom mycelia. The inhibitory activity on the growth of the mushroom fruiting bodies was evaluated by the treatment of the bacterial culture broth or the heat-treated cell-free supernatant of the broth. The treatment of the culture broths or the cell-free supernatants of LBS4 or LBS9 completely inhibited the formation of the fruiting body, thereby suggesting that the inhibitory agent is a heat-stable compound. In the case of LBS5, only the bacterial cell-containing culture broth was capable of inhibiting the formation of the fruiting body, whereas the cell-free supernatant did not, which suggests that an inhibitory agent generated by LBS5 is a protein or a heat-labile chemical compound, potentially a fungal cell wall-degrading enzyme. The culture broth of LBS1 was not inhibitory. However, its cell-free supernatant was capable of inhibiting the formation of fruiting bodies. This indicates that LBS1 may produce an inhibitory heat-stable chemical compound which is readily degraded by its own secreted enzyme.

Effects of in vitro Culture Period of Reconstructed Embryos and Genetic Background of Feeder Cells on Establishment of Embryonic Stem Cells Derived from Somatic Cell Nuclear Transfer Blastocysts in Pigs

  • Han, Na Rae;Baek, Song;Lee, Yongjin;Lee, Joohyeong;Yun, Jung Im;Lee, Eunsong;Lee, Seung Tae
    • Journal of Animal Reproduction and Biotechnology
    • /
    • v.35 no.1
    • /
    • pp.86-93
    • /
    • 2020
  • The establishment of porcine embryonic stem cells (ESCs) from porcine somatic cell nuclear transfer (SCNT) blastocysts is influenced by in vitro culture day of porcine reconstructed embryo and feeder cell type. Therefore, the objective of the present study was to determine the optimal in vitro culture period for reconstructed porcine SCNT embryos and mouse embryonic fibroblast (MEF) feeder cell type for enhancing colony formation efficiency from the inner cell mass (ICM) of porcine SCNT blastocysts and their outgrowth. As the results, porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days showed significantly increased efficiency in the formation of colonies, compared to those for 7 days. Moreover, MEF feeder cells derived from outbred ICR mice showed numerically the highest efficiency of colony formation in blastocysts produced through in vitro culture of porcine SCNT embryos for 8 days and porcine ESCs with typical ESC morphology were maintained more successfully over Passage 2 on outbred ICR mice-derived MEF feeder cells than on MEF feeder cells derived from inbred C57BL/6 and hybrid B6CBAF1 mice. Overall, the harmonization of porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days and MEF feeder cells derived from outbred ICR mice will greatly contribute to the successful establishment of ESCs derived from porcine SCNT blastocysts.

Cloning of Superoxide Dismutase (SOD) Gene of Lily 'Marcopolo' and Expression in Transgenic Potatoes

  • Park, Ji-Young;Kim, Hyun-Soon;Youm, Jung-Won;Kim, Mi-Sun;Kim, Ki-Sun;Joung, Hyouk;Jeon, Jae-Heung
    • Journal of Applied Biological Chemistry
    • /
    • v.49 no.1
    • /
    • pp.1-7
    • /
    • 2006
  • Differential display reverse transcription PCR (DDRT-PCR) analysis was performed on lily 'Marcopolo' bulb scale for isolation of expressed genes during bulblet formation. Cu/Zn lily-superoxide dismutase (LSOD) of 872 bp gene, with ability to scavenge reactive oxygen in stress environment, was isolated. Northern blot analysis showed expression levels of LSOD maximized 12 days after bulblet formation. Ti plasmid vectors were constructed with sense and antisense expressions of LSOD gene and transformed into potato. Southern blot analysis of transgenic potatoes revealed different copies of T-DNA were incorporated into potato genome. In transgenic potatoes, lily SOD gene was overexpressed in sense lines and not in antisense lines. In native polyacrylamide gel electrophoresis analysis, additional engineered LSOD was detected in sense overexpressed transgenic line only. Transgenic potatoes were subjected to oxidative stress, such as herbicide methyl viologen (MV). Transgenic potato lines with sense orientation exhibited increased tolerance to MV, whereas in antisense lines exhibited decreased tolerance. In vitro tuberization of transgenic potato with sense orientation was promoted, but was inhibited in transgenic potato with antisense orientation.

Inhibitors of AKT Signaling Pathway and their Application

  • WONG, Chin Piow
    • Proceedings of the Plant Resources Society of Korea Conference
    • /
    • 2019.04a
    • /
    • pp.33-33
    • /
    • 2019
  • The AKT signaling pathway is a highly regulated cell signaling system that forms a network with other cell signaling pathways. Hence, the AKT signaling pathway mediates several important cellular functions that include cell survival, proliferation, cell migration, and et cetera. Irregularities that led overactive AKT signaling have been linked to many diseases such as cancer and metabolic-associated diseases. Hence, modulating the overactive AKT signaling pathway via inhibitor is a tantalizing prospect for treatment of cancer and metabolic-associated diseases. Two inhibitors of the AKT signaling pathway will be presented in this symposium: 1) Bisleuconothine A (BisA), a bisindole alkaloid that inhibit autophagy and 2) Ceramicine B (CerB), a limonoid that inhibit adipogenesis. The first topic is on a bisindole alkaloid, BisA and its mechanism in inducing autophagosome formation in lung cancer cell line, A549.(1) Since most autophagy inducing agents generally induce apoptosis, we found that BisA does not induce apoptosis even in high dose. BisA up-regulation of LC3 lipidation is achieved through mTOR inactivation. The phosphorylation of PRAS40, a mTOR repressor was suppressed by BisA. This observation suggested that BisA inactivates mTOR via suppression of PRAS40 phosphorylation. Interestingly, the phosphorylation of AKT, an upstream regulator of PRAS40 phosphorylation was also down-regulated by BisA. These findings suggested that Bis-A induces autophagosomes formation by interfering with the AKT-mTOR signaling pathway. The second topic is on CerB and its mechanism in inhibiting adipogenesis in preadipocytes cell line, MC3T3-G2/PA6.(2,3) CerB inhibits the phosphorylation of protein kinase B (AKT) at the Thr308 position but not the Ser473. Consequently, the phosphorylation of FOXO3 which is located downstream of AKT is also inhibited. Considering that FOXO3 is an important regulator of PPARγ which is a key factor in adipogenesis, CerB may inhibit adipogenesis via the AKT-FOXO3 signaling pathway. Taken together, both BisA and CerB highlighted the potential of AKT signaling pathway modulation as an approach to induce autophagy and inhibit the formation of fat cells, respectively.

  • PDF

Teratoma Formation in Immunocompetent Mice After Syngeneic and Allogeneic Implantation of Germline Capable Mouse Embryonic Stem Cells

  • Aldahmash, Abdullah;Atteya, Muhammad;Elsafadi, Mona;Al-Nbaheen, May;Al-Mubarak, Husain Adel;Vishnubalaji, Radhakrishnan;Al-Roalle, Ali;Al-Harbi, Suzan;Manikandan, Muthurangan;Matthaei, Klaus Ingo;Mahmood, Amer
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.10
    • /
    • pp.5705-5711
    • /
    • 2013
  • Background: Embryonic stem cells (ESCs) have the potential to form teratomas when implanted into immunodeficient mice, but data in immunocompetent mice are limited. We therefore investigated teratoma formation after implantation of three different mouse ESC (mESC) lines into immunocompetent mice. Materials and Methods: BALB/c mice were injected with three highly germline competent mESCs (129Sv, BALB/c and C57BL/6) subcutaneously or under the kidney capsule. After 4 weeks, mice were euthanized and examined histologically for teratoma development. The incidence, size and composition of teratomas were compared using Pearson Chi-square, t-test for dependent variables, one-way analysis of variance and the nonparametric Kruskal-Wallis analysis of variance and median test. Results: Teratomas developed from all three cell lines. The incidence of formation was significantly higher under the kidney capsule compared to subcutaneous site and occurred in both allogeneic and syngeneic mice. Overall, the size of teratoma was largest with the 129Sv cell line and under the kidney capsule. Diverse embryonic stem cell-derived tissues, belonging to the three embryonic germ layers, were encountered, reflecting the pluripotency of embryonic stem cells. Most commonly represented tissues were nervous tissue, keratinizing stratified squamous epithelium (ectoderm), smooth muscle, striated muscle, cartilage, bone (mesoderm), and glandular tissue in the form of gut- and respiratory-like epithelia (endoderm). Conclusions: ESCs can form teratomas in immunocompetent mice and, therefore, removal of undifferentiated ESC is a pre-requisite for a safe use of ESC in cell-based therapies. In addition the genetic relationship of the origin of the cell lines to the ability to transplant plays a major role.

Electrical Switching Characteristics of Ge-Se Thin Films for ReRAM Cell Applications

  • Kim, Jang-Han;Nam, Ki-Hyun;Chung, Hong-Bay
    • Proceedings of the Korean Vacuum Society Conference
    • /
    • 2012.08a
    • /
    • pp.343-344
    • /
    • 2012
  • It has been known since the mid 1960s that Ag can be photodissolved in chalcogenide glasses to form materials with interesting technological properties. In the 40 years since, this effect has been used in diverse applications such as the fabrication of relief images in optical elements, micro photolithographic schemes, and for direct imaging by photoinduced Ag surface deposition. ReRAM, also known as conductive bridging RAM (CBRAM), is a resistive switching memory based on non-volatile formation and dissolution of a conductive filament in a solid electrolyte. Especially, Ag-doped chalcogenide glasses and thin films have become attractive materials for fundamental research of their structure, properties, and preparation. Ag-doped chalcogenide glasses have been used in the formation of solid electrolyte which is the active medium in ReRAM devices. In this paper, we investigated the nature of thin films formed by the photo-dissolution of Ag into Ge-Se glasses for use in ReRAM devices. These devices rely on ion transport in the film so produced to create electrically programmable resistance states. [1-3] We have demonstrated functionalities of Ag doped chalcogenide glasses based on their capabilities as solid electrolytes. Formation of such amorphous systems by the introduction of Ag+ ions photo-induced diffusion in thin chalcogenide films is considered. The influence of Ag+ ions is regarded in terms of diffusion kinetics and Ag saturation is related to the composition of the hosting material. Saturated Ag+ ions have been used in the formation of conductive filaments at the solid electrolyte which is the active medium in ReRAM devices. Following fabrication, the cell displays a metal-insulator-metal structure. We measured the I-V characteristics of a cell, similar results were obtained with different via sizes, due to the filamentary nature of resistance switching in ReRAM cell. As the voltage is swept from 0 V to a positive top electrode voltage, the device switches from a high resistive to a low resistive, or set. The low conducting, or reset, state can be restored by means of a negative voltage sweep where the switch-off of the device usually occurs.

  • PDF

Scale Formation by Electrode Reactions in Capacitive Deionization and its Effects on Desalination Performance (축전식 탈염에서 전극반응에 의한 스케일 생성과 탈염성능에 미치는 영향)

  • Choi, Jae-Hwan;Kang, Hyun-Soo
    • Applied Chemistry for Engineering
    • /
    • v.27 no.1
    • /
    • pp.74-79
    • /
    • 2016
  • The effects of scale formation of hardness material caused by electrode reactions on the desalination performance of the membrane capacitive deionization (MCDI) were investigated. During the repeated adsorption and desorption process for the influent containing $Ca^{2+}$ ion, changes in effluent concentration and cell potential with respect to the number of adsorption were analyzed. It was found that $OH^-$ generation at the cathode was initiated at about 0.8 V or more of cell potential. In addition, the scale of $Ca(OH)_2$ was formed on the surface of cathode carbon electrode by combining adsorbed $Ca^{2+}$ ions and $OH^-$ ions generated from electrode reaction. As the scale was forming, the electrical resistance of carbon electrode was increasing, which resulted in the decrease of the adsorption amount. In the case of the operation at 1.5 V cell potential, the adsorption was reduced to 58% of the initial adsorption amount due to the scale formation.

Formation of DNA-protein Cross-links Mediated by C1'-oxidized Abasic Lesion in Mouse Embryonic Fibroblast Cell-free Extracts

  • Sung, Jung-Suk;Park, In-Kook
    • Animal cells and systems
    • /
    • v.9 no.2
    • /
    • pp.79-85
    • /
    • 2005
  • Oxidized abasic residues arise as a major class of DNA damage by a variety of agents involving free radical attack and oxidation of deoxyribose sugar components. 2-deoxyribonolactone (dL) is a C1'-oxidized abasic lesion implicated in DNA strand scission, mutagenesis, and covalent DNA-protein cross-link (DPC). We show here that mammalian cell-free extract give rise to stable DPC formation that is specifically mediated by dL residue. When a duplex DNA containing dL at the site-specific position was incubated with cell-free extracts of Po ${\beta}-proficient$ and -deficient mouse embryonic fibroblast cells, the formation of major dL-mediated DPC was dependent on the presence of DNA polymerase (Pol) ${\beta}$. Formation of dL-specific DPC was also observed with histones and FEN1 nuclease, although the reactivity in forming dL-mediated DPC was significantly higher with Pol ${\beta}$ than with histones or FEN1. DNA repair assay with a defined DPC revealed that the dL lesion once cross-linked with Pol ${\beta}$ was resistant to nucleotide excision repair activity of cell-free extract. Analysis of nucleotide excision repair utilizing a model DNA substrate containing a (6-4) photoproduct suggested that excision process for DPC was inhibited because of DNA single-strand incision at 5' of the lesion. Consequently DPC mediated by dL lesion may not be readily repaired by DNA excision repair pathway but instead function as unusual DNA damage causing a prolonged DNA strand break and trapping of the major base excision repair enzyme.

Promoting Effect of Hydrogen Peroxide on 1-Methyl-4-phenylpyridinium-induced Mitochondrial Dysfunction and Cell Death in PC12 Cells

  • Lee, Dong-Hee;Lee, Chung-Soo
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.10 no.1
    • /
    • pp.51-58
    • /
    • 2006
  • The promoting effect of hydrogen peroxide ($H_2O_2$) against the cytotoxicity of 1-methyl-4-phenylpyridinium ($MPP^+$) in differentiated PC12 cells was assessed by measuring the effect on the mitochondrial membrane permeability. Treatment of PC12 cells with $MPP^+$ resulted in the nuclear damage, decrease in the mitochondrial transmembrane potential, cytosolic accumulation of cytochrome c, activation of caspase-3, increase in the formation of reactive oxygen species (ROS) and depletion of GSH. Addition of $H_2O_2$ enhanced the $MPP^+-induced$ nuclear damage and cell death. Catalase, Carboxy-PTIO, Mn-TBAP, N-acetylcysteine, cyclosporin A and trifluoperazine inhibited the cytotoxic effect of $MPP^+$ in the presence of $H_2O_2$. Addition of $H_2O_2$ promoted the change in the mitochondrial membrane permeability, ROS formation and decrease in GSH contents due to $MPP^+$ in PC12 cells. The results show that the $H_2O_2$ treatment promotes the cytotoxicity of $MPP^+$ against PC12 cells. $H_2O_2$ may enhance the $MPP^+$-induced viability loss in PC12 cells by promoting the mitochondrial membrane permeability change, release of cytochrome c and subsequent activation of caspase-3, which is associated with the increased formation of ROS and depletion of GSH. The findings suggest that $H_2O_2$ as a promoting agent for the formation of mitochondrial permeability transition may enhance the neuronal cell injury caused by neurotoxins.

Improvement of osteogenic potential of biphasic calcium phosphate bone substitute coated with synthetic cell binding peptide sequences

  • Choi, Hyunmin;Park, Nho-Jae;Jamiyandorj, Otgonbold;Hong, Min-Ho;Oh, Seunghan;Park, Young-Bum;Kim, Sungtae
    • Journal of Periodontal and Implant Science
    • /
    • v.42 no.5
    • /
    • pp.166-172
    • /
    • 2012
  • Purpose: The aim of this study was to evaluate the improvement of osteogenic potential of biphasic calcium phosphate (BCP) bone substitute coated with synthetic cell-binding peptide sequences in a standardized rabbit sinus model. Methods: Standardized 6-mm diameter defects were created bilaterally on the maxillary sinus of ten male New Zealand white rabbits, receiving BCP bone substitute coated with synthetic cell binding peptide sequences on one side (experimental group) and BCP bone substitute without coating (control group) on the other side. Histologic and histomorphometric analysis of bone formation was carried out after a healing period of 4 or 8 weeks. Results: Histological analysis revealed signs of new bone formation in both experimental groups (4- and 8-week healing groups) with a statistically significant increase in bone formation in the 4-week healing group compared to the control group. However, no statistically significant difference in bone formation was found between the 8-week healing group and the control group. Conclusions: This study found that BCP bone substitute coated with synthetic cell-binding peptide sequences enhanced osteoinductive potential in a standardized rabbit sinus model and its effectiveness was greater in the 4-week healing group than in the 8-week healing group.