• Title/Summary/Keyword: CELL/NATURE/SCIENCE

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Expression Profiling of Genes involved in the Control of Pluripotency Using cDNA Microarray

  • Lee, Young-Jin;Hong, Seok-Ho;Nah, Hee-Young;Chae, Ji-Hyung;Jung, Ho-Sun;Kim, Beom-Sue;Kim, Chul-Geun
    • Proceedings of the KSAR Conference
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    • 2001.10a
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    • pp.18-24
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    • 2001
  • To identify genes implicated in the control of pluripotency as well as characteristics of stem cells, we analyzed expression profiles of genes derived from mouse morulas, blastocysts, embryonic stem cells, mesenchymal stem cells, and uterus tissue using cDNA microarray. Comparative analyses of their expression profiles identified putative clones that expressed specifically in specific samples or not in a specific sample. The expression pattern of these candidate clones was analyzed using RT-PCR and non-radioactive in situ hybridization. Functional annotation of these clones on pluripotency and stem cell plasticity is in ongoing. These studies may further our understanding on the nature of the stem cells and molecular mechanisms underlying many facets of mammalian development and differentiation.

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Development of Polymeric Nanopaclitaxel and Comparison with Free Paclitaxel for Effects on Cell Proliferation of MCF-7 and B16F0 Carcinoma Cells

  • Yadav, Deepak;Anwar, Mohammad Faiyaz;Garg, Veena;Kardam, Hemant;Beg, Mohd Nadeem;Suri, Suruchi;Gaur, Sikha;Asif, Mohd
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2335-2340
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    • 2014
  • Paclitaxel is hydrophobic in nature and is recognized as a highly toxic anticancer drug, showing adverse effects in normal body sites. In this study, we developed a polymeric nano drug carrier for safe delivery of the paclitaxel to the cancer that releases the drug in a sustained manner and reduces side effects. N-isopropylacrylamide/vinyl pyrrolidone (NIPAAm/VP) nanoparticles were synthesized by radical polymerization. Physicochemical characterization of the polymeric nanoparticles was conducted using dynamic light scattering, transmission electron microscopy, scanning electron microscopy and nuclear magnetic resonance, which confirmedpolymerization of formulated nanoparticles. Drug release was assessed using a spectrophotometer and cell viability assays were carried out on the MCF-7 breast cancer and B16F0 skin cancer cell lines. NIPAAm/VP nanoparticles demonstrated a size distribution in the 65-108 nm range and surface charge measured -15.4 mV. SEM showed the nanoparticles to be spherical in shape with a slow drug release of ~70% in PBS at $38^{\circ}C$ over 96 h. Drug loaded nanoparticles were associated with increased viability of MCF-7 and B16F0 cells in comparison to free paclitaxel. Nano loaded paclitaxel shows high therapeutic efficiency by sustained release action for the longer period of time, i increasing its efficacy and biocompatibility for human cancer therapy. Therefore, paclitaxel loaded (NIPAAm/VP) nanoparticles may provide opportunities to expand delivery of the drug for clinical selection.

Examination of Cytopathic Effect and Apoptosis in Listeria monocytogenes-Infected Hybridoma B-Lymphocyte (Ped-2E9) Line In Vitro

  • Bhunia, Arun-Kumar;Feng, Xiang
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.398-403
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    • 1999
  • In our previous studies, we reported that hybridoma B-lymphocytes can be used to determine the virulence of Listeria species in an in vitro cytotoxicity assay. Here, we examined the cytopathic effect, i.e., membrane damage and the nature of cell death induced by Listeria monocytogenes on murine hybridoma B-lymphocytes (Ped-2E9). Membrane damage was assessed by microscopic analyses and by measuring the release of intracellular alkaline phosphatase(AP) and lactate dehydrogenase (LDH). Cell death was determined by DNA fragmentation analyses using agarose gel electrophoresis. Infection by listeriolysin O (LLO)-producing L. monocytogenes strains induced substantial amounts of AP and LDH release from Ped-2E9 hybridoma B-cells, suggesting severe membrane damage in these cells, while an LLO-negative L. monocytogenes mutant strain had no effect. An LLO-producing recombinant L. innocua ($prifA^+hly^+$) strain also induced high AP and LDH release and cytopathic changes in Ped-2E9 cells. Light or scanning electron microscopic examination revealed L. monocytogenes mediated membrane destabilization, pore formation, intense cytoplasmic granulation, bleb formation, and lysis of Ped-2E9 cells. LLO-producing L. monocytogenes and L. innocua ($prifA^{+}hly{^}+$) also induced ladder-like DNA fragmentation in Ped-2E9 cells. Collectively, these results suggest that L. monocytogenes, specifically LLO-producing strains, can induce a severe cytopathic effect leading to apoptosis in hybridoma B-lymphocytes (Ped-2E9).

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Screening of Indigenous Strains of Lactic Acid Bacteria for Development of a Probiotic for Poultry

  • Karimi Torshizi, M.A.;Rahimi, Sh.;Mojgani, N.;Esmaeilkhanian, S.;Grimes, J.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.10
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    • pp.1495-1500
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    • 2008
  • In an attempt to develop a probiotic formulation for poultry feed, a number of lactic acid bacteria (LAB) were isolated from chicken intestinal specimens and a series of in vitro experiments were performed to evaluate their efficacy as a potential probiotic candidate. A total of 650 LAB strains were isolated and screened for their antagonistic potential against each other. Among all the isolates only three isolates (TMU121, 094 and 457) demonstrated a wide spectrum of inhibition and were thus selected for detailed investigations. All three selected isolates were able to inhibit the growth of E. coli and Salmonella species, although to variable extent. The nature of the inhibitory substance produced by the isolates TMU121 and 094 appeared to be associated with bacteriocin, as their activity was completely lost after treatment with proteolytic enzymes, while pH neutralization and catalase enzyme had no effect on the residual activity. In contrast, isolate TMU457 was able to resist the effect of proteolytic enzymes while pH neutralization completely destroyed its activity. Attempts were made to study the acid, bile tolerance and cell surface hydrophobicity of these isolates. TMU121 showed high bile salt tolerance (0.3%) and high cell surface hydrophobicity compared to the other two strains studied, while TMU094 appeared the most pH resistant strain. Based on these results, the three selected LAB isolates were considered as potential ingredients for a chicken probiotic feed formulation and were identified to species level based on their carbohydrate fermentation pattern by using API 50CH test kits. The three strains were identified as Lactobacillus fermentum TMU121, Lactobacillus rhamnosus TMU094, and Pediococcus pentosaceous TMU457.

Evidence That Temporally Alternative Expression of the Vibrio vulnificus Elastase Prevents Proteolytic Inactivation of Hemolysin

  • Rhee, Jee-Eun;Lee, Jeong-Hyun;Jeong, Hye-Sook;Park, U-Ryung;Lee, Dong-Ha;Woo, Gun-Jo;Miyoshi, Shin-Ichi;Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.1021-1026
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    • 2003
  • Numerous secreted and cell-associated virulence factors have been proposed to account for the fulminating and destructive nature of Vibrio vulnificus infections. Among the putative virulence factors are an elastase, elastolytic protease, and a cytolytic hemolysin. Effects of the elastase on the hemolysin were assessed by evaluating changes of hemolytic activities either in the presence or absence of the protease. Although hemolytic activity in the culture supernatant was lowered by the purified elastase added in vitro, the cellular level of hemolytic activity was unaffected by the mutation of vvpE encoding the elastase. Growth kinetic studies revealed that hemolysin reached its maximum level in the exponential phase of growth, and the elastase appeared at the onset of the stationary phase. These results have provided insight into the regulation of virulence factors: temporally coordinate regulation of virulence factors is essential for the overall success of the pathogen during pathogenesis.

Cytochemical Localization of Nuclear Actin of Sperm and Spermatids in Urechis unicinctus

  • Shin, Kil-Sang;Kim, Ho-Jin;Kwon, Hyuk-Jae;Kim, Wan-Jong
    • Animal cells and systems
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    • v.9 no.2
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    • pp.65-73
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    • 2005
  • In this study, we found that sperm ball of Urechis unicinctus consisted of a somatic cell and spermatogenic cells. After separation from the sperm ball, individual spermatid floated freely in the coelomic fluid and differentiated into a mature sperm. Because of many nuclear vacuoles, spermatid nucleus was observed to be heterogeneous. Later, the spermatid nucleus condensed into the homogeneous round nucleus of the mature sperm. Perinuclear microtubules could be seen but did not seem to be organized into manchette microtubules. To understand the nature of nuclear condensation during spermiogenesis, the sperm and spermatids (spermiogenic cells) were treated with FITC-phalloidin, or anti-actin-FITC, or labeled with antiactin immunogold particles (AAIP; 10 nm) followed by transmission electron microscopy or confocal laser scanning microscopy. The anti-actin-FITC and FITC-phalloidin reactions occurred distinctly in the nuclei of both spermiogenic cells. FITC-phalloidin reacted more intensely with acrosomes. The AAIP were incorporated mainly into nuclei of both cells sometimes showing local distribution in the nucleus. Nuclear vacuoles of spermatids disappeared progressively with condensation of the nucleus, as the number of incorporated $AAIP/{\mu}m^2$ increased. These results suggest that nuclear actin microfilaments might be closely related to nuclear condensation.

Artificial Reestablishment of the Kelp and Red Algal Symbiosis

  • Kwang Young Kim;David J. Garbary
    • Animal cells and systems
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    • v.3 no.3
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    • pp.243-246
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    • 1999
  • A type of symbiosis was previously described from nature in which the gametophytes of Laminariales were endophytic in filamentous red algae. Here we reconstruct this symbiosis for the first time in laboratory culture using zoospores of the kelp, Undaria pinnatifida, and the red alga, Aglaothamnion oosumiense. Zoospores of U. pinnatifida readily attached to A. oosumiense. In 48 h these spores germinated and the initial germ tube penetrated into the host cell wall leaving only an empty zoospore wall outside the host. Within ten days, four to five-celled endophytic gametophytes were present. Zoospores of Laminaria religiosa which were also inoculated into cultures of A. oosumiense rarely attached to the red alga and never became endophytic. Within ten days the free-living gametophytes of L. religiosa on cover slips became fertile and produced young sporophytes. These observations demonstrate the ability of U. pinnatifida to become endophytic, and show differences in host specificity among kelp species.

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In vitro Constructive Approaches to the Origin of Coding Sequences

  • Shiba, Kiyotaka
    • BMB Reports
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    • v.31 no.3
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    • pp.209-220
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    • 1998
  • How did nature create the first set of genes at the beginning of life on Earth? One of the goals of molecular biology is to elucidate the fundamental rules governing how genes and, therefore, proteins were created. Through experiments carried out in the emerging field of "in vitro" or "benchtop" evolution studies, we are gaining new insights into the origins of genes and proteins as well as the origins of their functions (e.g., catalysis). In this review, I present an overview of recent experimental approaches to the question of the origin and evolution of genes. In addition, I will introduce a novel in vitro protein emergence system that was recently developed in my laboratory.

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Analysis of the Singular Point of Cyclic Voltammograms Recorded with Various Scan Rates

  • Chang, Byoung-Yong
    • Journal of Electrochemical Science and Technology
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    • v.8 no.3
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    • pp.244-249
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    • 2017
  • This paper presents the results of an investigation into the isoamperic point of cyclic voltammograms, which is defined as the singular point where the voltammograms of various scan rates converge. The origin of the unique point is first considered from a theoretical perspective by formulating the voltammetric curves as a system of linear equations, the solution of which indicates that a trivial solution is only available at the potential at which the net current is zero during the reverse potential scan. In addition, by way of a mathematical formulation, it was also shown that the isoamperic point is dependent on the switching potential of the potential scanning. To validate these findings, theoretical and practical cyclic voltammmograms were studied using finite-element based digital simulations and 3-electrode cell experiments. The new understanding of the nature of the isoamperic point provides an opportunity to measure the charge transfer effects without the influence of the mass transfer effects when determining the thermodynamic and kinetic characteristics of a faradaic system.

Simulating astrophysical shocks with a combined PIC MHD code

  • van Marle, Allard Jan
    • The Bulletin of The Korean Astronomical Society
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    • v.43 no.1
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    • pp.56.2-56.2
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    • 2018
  • Astrophysical shocks accelerate particles to high velocities, which we observe as cosmic rays. The acceleration process changes the nature of the shock because the particles interact with the local magnetic field, removing energy and potentially triggering instabilities. In order to simulate this process, we need a computational method that can handle large scale structures while, at the same time, following the motion of individual particles. We achieve this by combining the grid magnetohydrodynamics (MHD) method with the particle-in-cell (PIC) approach. MHD can be used to simulate the thermal gas that forms the majority of the gas near the shock, while the PIC method allows us to model the interactions between the magnetic field and those particles that deviate from thermal equilibrium. Using this code, we simulate shocks at various sonic and Alfvenic Mach numbers in order to determine how the behaviour of the shock and the particles depends on local conditions.

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