• 제목/요약/키워드: cellular metals

검색결과 68건 처리시간 0.026초

Identification of Differentially Expressed Genes by Exposure of Methylmercury in Neuroblastoma Cell Line Using Suppression Subtractive Hybridization (SSH)

  • Kim, Youn-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • 제2권1호
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    • pp.60-66
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    • 2006
  • Methylmercury (MeHg), one of the heavy metal compounds, can cause severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. This study, using of suppression subtractive hybridization (SSH) method, was peformed to identify differentially expressed genes by MeHg in SH-SY5Y human neuroblastoma cell line. We prepared to total RNA from SH-SY5Y cells treated with solvent (DMSO) and $6.25\;{\mu}M\;(IC_{50})$ MeHg and performed forward and reverse SSH. Differentially expressed cDNA clones were screened by dot blot, sequenced and confirmed that individual clones indeed represent differentially expressed genes with real time RT-PCR. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences may provide an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as ubiquitous environmental pollutants.

멕시코 B 광산 슬러지에 존재하는 생물학적 광물화 미생물의 특성에 관한 연구 (Isolation and Identification of Bacteria Involved with Biomineralization at B Mine Sludge in Mexico)

  • 김준하;윤성열;박윤수;이재영
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제22권2호
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    • pp.41-51
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    • 2017
  • Microbial processes that bind heavy metals and form minerals are widespread, and they represent a basic aspect of biogeochemistry. Some microorganisms can crystallize minerals by secreting a specific enzyme. In particular, calcite ($CaCO_3$) precipitation is an important part of biomineralization, and has been studied extensively because of its wide application in civil engineering technology. This process provides an effective way to stabilize heavy metals within a relatively stable crystal phase. In this study, biomineralization of calcite by three urea-hydrolyzing indigenous bacterial strains was investigated by microbiological analyses. Three bacterial strains were isolated from the sludge of B mine in Mexico and each bacterial strain was identified by the cellular fatty acid composition and 16S rRNA partial sequencing analysis. The results of the identification analysis showed that these strains were closest to Sporosarcina pasteurii, Kurthia gibsonii, and Paenibacillus polymyxa. We found that the optimum conditions for growth of these indigenous bacteria were $30-40^{\circ}C$ and pH range of 7-8. Microbiological analyses showed the possibility that the bioaccumulated heavy metals ions were deposited around the cell as crystalline carbonate minerals under the optimum conditions. The findings of our study suggest that the indigenous bacterial strains play an important role in heavy metal immobilization.

Biochemical and Biodiversity Insights into Heavy Metal Ion-Responsive Transcription Regulators for Synthetic Biological Heavy Metal Sensors

  • Jung, Jaejoon;Lee, Sang Jun
    • Journal of Microbiology and Biotechnology
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    • 제29권10호
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    • pp.1522-1542
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    • 2019
  • To adapt to environmental changes and to maintain cellular homeostasis, microorganisms adjust the intracellular concentrations of biochemical compounds, including metal ions; these are essential for the catalytic function of many enzymes in cells, but excessive amounts of essential metals and heavy metals cause cellular damage. Metal-responsive transcriptional regulators play pivotal roles in metal uptake, pumping out, sequestration, and oxidation or reduction to a less toxic status via regulating the expression of the detoxification-related genes. The sensory and regulatory functions of the metalloregulators have made them as attractive biological parts for synthetic biology, and the exceptional sensitivity and selectivity of metalloregulators toward metal ions have been used in heavy metal biosensors to cope with prevalent heavy metal contamination. Due to their importance, substantial efforts have been made to characterize heavy metal-responsive transcriptional regulators and to develop heavy metal-sensing biosensors. In this review, we summarize the biochemical data for the two major metalloregulator families, SmtB/ArsR and MerR, to describe their metal-binding sites, specific chelating chemistry, and conformational changes. Based on our understanding of the regulatory mechanisms, previously developed metal biosensors are examined to point out their limitations, such as high background noise and a lack of well-characterized biological parts. We discuss several strategies to improve the functionality of the metal biosensors, such as reducing the background noise and amplifying the output signal. From the perspective of making heavy metal biosensors, we suggest that the characterization of novel metalloregulators and the fabrication of exquisitely designed genetic circuits will be required.

Modified Suppression Subtractive Hybridization Identifies an AP2-containing Protein Involved in Metal Responses in Physcomitrella patens

  • Cho, Sung Hyun;Hoang, Quoc Truong;Phee, Jeong Won;Kim, Yun Young;Shin, Hyun Young;Shin, Jeong Sheop
    • Molecules and Cells
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    • 제23권1호
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    • pp.100-107
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    • 2007
  • The moss Physcomitrella patens has two life cycles, filamentous protonema and leafy gametophore. A modified from of suppression subtractive hybridization (SSH), mirror orientation selection (MOS), was applied to screen genes differentially expressed in the P. patens protonema. Using reverse Northern blot analysis, differentially expressed clones were identified. The identified genes were involved mainly in metal binding and detoxification. One of these genes was an AP2 (APETALA2) domain-containing protein (PpACP1), which was highly up-regulated in the protonema. Alignment with other AP2/EREBPs (Ethylene Responsive Element Binding Proteins) revealed significant sequence homology of the deduced amino acid sequence in the AP2/EREBP DNA binding domain. Northern analysis under various stress conditions showed that PpACP1 was induced by ethephon, cadmium, copper, ABA, IAA, and cold. In addition, it was highly expressed in suspension-cultured protonema. We suggest that PpACP1 is involved in responses to metals, and that suspension culture enhance the expression of genes responding to metals.

Subcellular Distribution of Heavy Metals in Organs of Bivalve Modiolus Modiolus Living Along a Metal Contamination Gradient

  • Podgurskaya, Olga V.;Kavun, Victor Ya.
    • Ocean Science Journal
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    • 제41권1호
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    • pp.43-51
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    • 2006
  • Concentration and distribution of Fe, Zn, Cu, Cd, Mn, Pb, Ni among subcellular fractions (cellular membrane structures and cytosol) and Zn, Cu, Cd among cytoplasmic proteins in the kidney and digestive gland of mussel Modiolus modiolus living along a polymetallic concentration gradient were studied. It was found in the kidney of M. modiolus from contaminated sites that the Fe percent increased in the "membrane" fraction, whereas Zn, Pb, Ni and Mn percent increased in the cytosol compared to the kidney of the control mussel. Note kidney cytosol of M. modiolus from clean and contaminated sites sequestered major parts of Cu and Cd. In the digestive gland of M. modiolus from contaminated sites Fe, Zn, Cd, Mn, Ni percent increased in the "membrane" fraction, whereas Cu, Pb percent increased in the cytosol compared to digestive gland of control mussel. Gel-filtration chromatography shows kidney of M. modiolus contains increased metallothionein-like protein levels irrespective of ambient dissolved metal concentrations. It was shown that the metal detoxification system in the kidney and digestive gland of M. modiolus was efficient under extremely high ambient metal levels. However, under complex environmental contamination in the kidney of M. modiolus, the metal detoxification capacity of metallothionein-like proteins was damaged.

다이아몬드 트러스 벽면으로 구성된 P-TDC 모델의 강성 및 강도 연구 (Study of Effective Stiffness and Effective Strength for a Pinwheel Model combined with Diamond Truss-Wall Corrugation (P-TDC))

  • 최정호
    • 한국산업융합학회 논문집
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    • 제19권3호
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    • pp.109-124
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    • 2016
  • The objective of this paper is to find the density, stiffness, and strength of truss-wall diamond corrugation model combined with pinwheel truss inside space. The truss-wall diamond corrugation (TDC) model is defined as a unit cell coming from solid-wall diamond corrugation (SDC) model. Pinwheel truss-wall diamond corrugation (P-TDC) model is made by TDC connected with pinwheel structure inside of the space. Derived ideal solutions of P-TDC is based on truss-wall and pinwheel truss model at first. And then it is compared with Gibson-Ashby's ideal solution. To validate the ideal solutions of the P-TDC, ABAQUS software is used to predict the density, strength, and stiffness, and then each of them are compared to the ideal solution of Gibson-Ashby with a log-log scale. Applied material property is stainless steel 304 because of having cost effectiveness. Applied parameters for P-TDC are 1 thru 5 mm diameter within fixed opening width as 4mm. In conclusion, the relative Young's modulus and relative yield strength of the P-TDC unit model is reasonable matched to the ideal expectations of the Gibson-Ashby's theory. In nearby future, P-TDC model is hoped to be applied to make sandwich core structure by advanced technologies such as 3D printing skills.

Wire-woven Bulk Kagome의 압축 특성 분석 (Analysis of Compressive Characteristics of Wire-woven Bulk Kagome)

  • 이병곤;최지은;강기주;전인수
    • 대한기계학회논문집A
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    • 제32권1호
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    • pp.70-76
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    • 2008
  • Periodic cellular metals (PCMs) are actively being investigated because of their excellent specific strength and stiffness, and multi-functionality such as a heat disperse structure bearing external loading. The Kagome truss PCM has been proved that it has higher resistance to plastic buckling and lower anisotropy than other truss PCMs. In this paper, the out-of-plane compressive responses of the WBK specimens have been measured, theoretically predicted and numerically analyzed. Three specimens of two-layered WBK are fabricated and tested for measuring the responses. The peak stress of compressive behavior and effective elastic modulus are predicted based on the equilibrium equation and elastic energy conservation. Moreover, the structure of the specimen is modeled using the commercial mesh generation code, PATRAN and the finite element analysis for the model under the compression is carried out using the commercial FE code, ABAQUS. Finally, the obtained results are compared with each other to analyze the compressive characteristics of Wire-woven Bulk Kagome (WBK).

Effect of Copper on Marine Microalga Tetraselmis suecica and its Influence on Intra- and Extracellular Iron and Zinc Content

  • Kumar, K. Suresh;Shin, Kyung-Hoon
    • 생태와환경
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    • 제50권1호
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    • pp.16-28
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    • 2017
  • In an aquatic environment, toxicity of metals to organisms depends on external factors (type of metal, exposure concentration and duration, environmental parameters, and water quality) and intracellular processes(metal-binding sites and detoxification). Toxicity of copper(Cu) on the marine microalga Tetraselmis suecica was investigated in this study. Dose-dependent (Cu concentration dependent) inhibition of growth and cell division, as well as, variation of intra- and extra-cellular Cu, Fe and Zn content was observed. T. suecica was sensitive to Cu; the 96 h $EC_{50}$ (concentration to inhibit growth-rate by 50%) of growth rate (${\mu}$) ($21.73{\mu}M\;L^{-1}$), cell division $day^{-1}$ ($18.39{\mu}M\;L^{-1}$), and cells $mL^{-1}$ ($13.25{\mu}M\;L^{-1}$) demonstrate the toxicity of Cu on this microalga. High intra-($19.86Pg\;cell^{-1}$) and extra-cellular($54.73Pg\;cell^{-1}$) Cu concentrations were recorded, on exposure to 24.3 and $72.9{\mu}M\;L^{-1}$ of Cu.

TATA-Binding Protein-Related Factor 2 Is Localized in the Cytoplasm of Mammalian Cells and Much of It Migrates to the Nucleus in Response to Genotoxic Agents

  • Park, Kyoung-ae;Tanaka, Yuji;Suenaga, Yusuke;Tamura, Taka-aki
    • Molecules and Cells
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    • 제22권2호
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    • pp.203-209
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    • 2006
  • TBP (TATA-binding protein)-related factor 2 (TRF2) regulates transcription during a nuber of cellular processes. We previously demonstrated that it is localized in the cytoplasm and is translocated to the nucleus by DNA-damaging agents. However, the cytoplasmic localization of TRF2 is controversial. In this study, we reconfirmed its cytoplasmic localization in various ways and examined its nuclear migration. Stresses such as heat shock, redox agents, heavy metals, and osmotic shock did not affect localization whereas genotoxins such as methyl methanesulfonate (MMS), cisplatin, etoposide, and hydroxyurea caused it to migrate to the nucleus. Adriamycin, mitomycin C and ${\gamma}$-rays had no obvious effect. We determined optimal conditions for the nuclear migration. The proportions of cells with nuclei enriched for TRF2 were 25-60% and 5-10% for stressed cells and control cells, respectively. Nuclear translocation was observed after 1 h, 4 h and 12 h for cisplatin, etoposide and MMS and hydroxyurea, respectively. The association of TRF2 with the chromatin and promoter region of the proliferating cell nuclear antigen (PCNA) gene, a putative target of TRF2, was increased by MMS treatment. Thus TRF2 may be involved in genotoxin-induced transcriptional regulation.

연속된 와이어를 이용한 다층 PCM의 제조방법 및 특성 평가 (A New Method to Fabricate Bulk PCMs from Continuous Wires and the Mechanical Behaviors)

  • 이용현;최지은;전인수;강기주
    • 대한기계학회논문집A
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    • 제31권2호
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    • pp.245-252
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    • 2007
  • Since the new millennium, truss PCMs(Periodic Cellular Metals) have drawn attention because of their superior specific stiffness, strength and multi-functionality. Prior studies have focused on the structural design and optimization. Kagome truss PCM has been proved to have the higher resistance to plastic buckling, more plastic deformation energy and lower anisotropy than other truss PCMs. In this study, we introduce a new idea to fabricate multi-layered Kagome truss PCM from continuous wires which can gain high strength as in piano wires and can be controlled to be defect free owing to drawing process. The relative density, the stiffness and the strength under bending and compressive load are estimated through elementary mechanics and compared with the results from experiments and FEA. The failure mechanisms are analyzed, and also mechanical performance and production are discussed.