• Title/Summary/Keyword: membrane processes

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Electrophysiological Characteristics of Six Mutations in hClC-1 of Korean Patients with Myotonia Congenita

  • Ha, Kotdaji;Kim, Sung-Young;Hong, Chansik;Myeong, Jongyun;Shin, Jin-Hong;Kim, Dae-Seong;Jeon, Ju-Hong;So, Insuk
    • Molecules and Cells
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    • v.37 no.3
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    • pp.202-212
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    • 2014
  • ClC-1 is a member of a large family of voltage-gated chloride channels, abundantly expressed in human skeletal muscle. Mutations in ClC-1 are associated with myotonia congenita (MC) and result in loss of regulation of membrane excitability in skeletal muscle. We studied the electrophysiological characteristics of six mutants found among Korean MC patients, using patch clamp methods in HEK293 cells. Here, we found that the autosomal dominant mutants S189C and P480S displayed reduced chloride conductances compared to WT. Autosomal recessive mutant M128I did not show a typical rapid deactivation of Cl- currents. While sporadic mutant G523D displayed sustained activation of $Cl^-$ currents in the whole cell traces, the other sporadic mutants, M373L and M609K, demonstrated rapid deactivations. $V_{1/2}$ of these mutants was shifted to more depolarizing potentials. In order to identify potential effects on gating processes, slow and fast gating was analyzed for each mutant. We show that slow gating of the mutants tends to be shifted toward more positive potentials in comparison to WT. Collectively, these six mutants found among Korean patients demonstrated modifications of channel gating behaviors and reduced chloride conductances that likely contribute to the physiologic changes of MC.

Radioactive cDNA microarray in Neurospsychiatry (신경정신 의학분야의 방사성동위원소 표지 cDNA 마이크로어레이)

  • Choe, Jae-Gol;Shin, Kyung-Ho;Lee, Min-Soo;Kim, Meyoung-Kon
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.1
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    • pp.43-52
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    • 2003
  • Microarray technology allows the simultaneous analysis of gene expression patterns of thousands of genes, in a systematic fashion, under a similar set of experimental conditions, thus making the data highly comparable. In some cases arrays are used simply as a primary screen loading to downstream molecular characterization of individual gene candidates. In other cases, the goal of expression profiling is to begin to identify complex regulatory networks underlying developmental processes and disease states. Microarrays were originally used with ceil lines or other simple model systems. More recently, microarrays have been used in the analysis of more complex biological tissues including neural systems and the brain. The application of cDNA arrays in neuropsychiatry has lagged behind other fields for a number of reasons. These include a requirement for a large amount of input probe RNA In fluorescent-glass based array systems and the cellular complexity introduced by multicellular brain and neural tissues. An additional factor that impacts the general use of microarrays in neuropsychiatry is the lack of availability of sequenced clone sets from model systems. While human cDNA clones have been widely available, high qualify rat, mouse, and drosophilae, among others are just becoming widely available. A final factor in the application of cDNA microarrays in neuropsychiatry is cost of commercial arrays. As academic microarray facilitates become more commonplace custom made arrays will become more widely available at a lower cost allowing more widespread applications. in summary, microarray technology is rapidly having an impact on many areas of biomedical research. Radioisotope-nylon based microarrays offer alternatives that may in some cases be more sensitive, flexible, inexpensive, and universal as compared to other array formats, such as fluorescent-glass arrays. In some situations of limited RNA or exotic species, radioactive membrane microarrays may be the most practical experimental approach in studying psychiatric and neurodegenerative disorders, and other complex questions in the brain.

A Study on Metal Concentrations in the Air of Metal Products Manufacturing Industry (금속제품 제조 산업장내 공기중 금속농도에 관한 연구)

  • Kang, Yong Seon;Kim, Se Dong;Ku, Tae Hyeong;Yoon, Hyeong Ryeol;Moon, Deog Hwan;Han, Yong Soo
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.6 no.2
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    • pp.249-264
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    • 1996
  • This study was conducted for the purpose of obtaining the fundamental data on improvement of working environment and contributing to health improvement of workers who dealed with metal by assessing the metal concentration in air of industries located in Chang-Won Industrial Complex. Authors measured the concentration of metals(Al, Cd, Cr, Cu, Mn, Ni, Pb, Sn and Zn) is the air to 25 working processes of 73 industries by flame atomic absorption spectrometry from February to December 1994. Personal air sampler was used for air sampling with mixed cellulose-ester membrane filter. The results were as follows : 1. The geometric means(range) of metal concentration; 1) Al: $0.1505mg/m^3$ ($0.0147-18.6100mg/m^3$) 2) Cd: $0.0077mg/m^3$ ($0.0003-7.0710mg/m^3$) 3) Cr: $0.0163mg/m^3$ ($0.0013-1.1510mg/m^3$) 4) Cu: $0.0097mg/m^3$ ($0.0009-0.4950mg/m^3$) 5) Mn: $0.0412mg/m^3$ ($0.0006-4.7877mg/m^3$) 6) Ni: $0.0088mg/m^3$ ($0.0001-1.0170mg/m^3$) 7) Pb: $0.0152mg/m^3$ ($0.0015-0.4499mg/m^3$) 8) Sn: $0.0486mg/m^3$ ($0.0037-0.1500mg/m^3$) 9) Zn: $0.1911mg/m^3$ ($0.0122-8.2920mg/m^3$) 2. The geometric mean of lead exceeded TWA in assembling process of other general purpose machinery not elsewhere classified products manufacturing industries.

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Improved Purification of Thermophilic FoF1-ATP Synthase c-Subunit Rings and Solid-State NMR Characterization of Them in Different Lipid Membranes

  • Bak, Suyeon;Kang, Su-Jin;Suzuki, Toshiharu;Yoshida, Masasuke;Fujiwara, Toshimichi;Akutsu, Hideo
    • Journal of the Korean Magnetic Resonance Society
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    • v.17 no.2
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    • pp.67-75
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    • 2013
  • ATP synthase produces ATP, a major energy source for metabolic processes in organisms, from ADP and inorganic phosphate in cellular membranes. ATP synthase is known as a rotary motor, in which the c-subunit ring functions as a rotor. In this work, we have tried to develop a more general preparation procedure of thermophilic $F_oc$-ring ($TF_oc$-ring) for NMR measurements. The expression of $TF_oF_1$ is easily affected by various experimental conditions such as temperature, shape and size of a flask, a volume of medium, and shaking rate of an incubator. Accordingly, we have tried to optimize the expression conditions of $TF_oF_1$. $TF_oc$-rings were purified from $TF_oF_1$ according to a reported method. We modified purification procedures to improve purity and yield of $TF_oc$. On top of them, we found a new combination of detergents for the purification at anion-exchange column chromatography. To examine the effect of lipid environments on the structure, the $TF_oc$-rings were reconstituted into two kinds of lipid bilayers, namely, saturated and unsaturated lipid ones. Then, we have compared characteristics of the $TF_oc$-ring structures in these membranes with solid-state NMR.

The application of chitosan to dental medicine

  • Hayashi, Y.;Yamada, S.;Ohara, N.;Kim, S-K.;Ikeda, T.;Yanagiguchi, K.;Matsunaga, T.
    • Proceedings of the KACD Conference
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    • 2003.11a
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    • pp.545-545
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    • 2003
  • Chitosan is applied as a dressing for oral mucous wound and a tampon following radical treatment of maxillary sinus. Furthermore, it is being investigated as an absorbing membrane for endodontic and periodontic surgeries. A few studies have reported osteoconduction and osteogenesia at the site of chitosan implant in vivo. However, compared with soft tissue healing processes, the mechanisms concerning effects of chitosan for biological mineralization have not yet been resoil In the present study, we studied the gene expression pattern using cDNA microarray and RT-PCR analyses in hard tissue forming osteoblasts cultured with water-soluble and low molecular weight chitooligosaccharide. cDNA microarray analysis revealed that 16 genes were expressed at 〉1.5-fold higher signal ratio levels in the experimental group compared with the control group after 3 days. RT-PCR analysis showed that chitosan oligomer induced an increase in the expression of two genes, CD56 antigen and tissue-type plasminogen activator. Furthermore, the expression of mRNAs for BMP-2 was almost identical in the experimental and control groups after 3 days of culture, but slightly increased after 7 days of culture with chitosan oligomer. These results suggest that a super-low concentration of chitooligosaccharide could modulate the activity of osteoblastic cells through mRNA levels and that the genes concerning cell proliferation and differentiation can be controlled by water-soluble chitosan.

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A Correlative Study on Aβ and CD95 Pathway Independent to Ca2+ Dependent Protease and Activation of Caspase Activation

  • Tuyet, Pham Thi Dieu
    • Journal of Integrative Natural Science
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    • v.7 no.1
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    • pp.25-38
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    • 2014
  • Amyloid-${\beta}$-peptide ($A{\beta}$) is important in the pathogenesis of Alzheimer's disease (AD). Calpain ($Ca^{2+}$-dependent protease) and caspase-8 (the initiating caspase for the extrinsic, receptor-mediated apoptosis pathway) have been implicated in $AD/A{\beta}$ toxicity. We found that $A{\beta}$ promoted degradation of calpastatin (the specific endogenous calpain inhibitor); calpastatin degradation was prevented by inhibitors of either calpain or caspase-8. The results implied a cross-talk between the two proteases and suggested that one protease was responsible for the activity of the other one. In neuron-like differentiated PC12 cells, calpain promotes active caspase-8 formation from procaspase-8 via the $A{\beta}$ and CD95 pathways, along with degradation of the procaspase-8 processing inhibitor caspase-8 (FLICE)-like inhibitory protein, short isoform (FLIPS). Inhibition of calpain (by pharmacological inhibitors and by overexpression of calpastatin) prevents the cleavage of procaspase-8 to mature, active caspase-8, and inhibits FLIPS degradation in the $A{\beta}$-treated and CD95-triggered cells. Increased cellular Ca2+ per se results in calpain activation but does not lead to caspase-8 activation or FLIPS degradation. The results suggest that procaspase-8 and FLIPS association with cell membrane receptor complexes is required for calpain-induced caspase-8 activation. The results presented here add to the understanding of the roles of calpain, caspase- 8, and CD95 pathway in $AD/A{\beta}$ toxicity. Calpain-promoted activation of caspase-8 may have implications for other types of CD95-induced cell damage, and for nonapoptotic functions of caspase-8. Inhibition of calpain may be useful for modulating certain caspase-8-dependent processes.

Consensus channelome of dinoflagellates revealed by transcriptomic analysis sheds light on their physiology

  • Pozdnyakov, Ilya;Matantseva, Olga;Skarlato, Sergei
    • ALGAE
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    • v.36 no.4
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    • pp.315-326
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    • 2021
  • Ion channels are membrane protein complexes mediating passive ion flux across the cell membranes. Every organism has a certain set of ion channels that define its physiology. Dinoflagellates are ecologically important microorganisms characterized by effective physiological adaptability, which backs up their massive proliferations that often result in harmful blooms (red tides). In this study, we used a bioinformatics approach to identify homologs of known ion channels that belong to 36 ion channel families. We demonstrated that the versatility of the dinoflagellate physiology is underpinned by a high diversity of ion channels including homologs of animal and plant proteins, as well as channels unique to protists. The analysis of 27 transcriptomes allowed reconstructing a consensus ion channel repertoire (channelome) of dinoflagellates including the members of 31 ion channel families: inwardly-rectifying potassium channels, two-pore domain potassium channels, voltage-gated potassium channels (Kv), tandem Kv, cyclic nucleotide-binding domain-containing channels (CNBD), tandem CNBD, eukaryotic ionotropic glutamate receptors, large-conductance calcium-activated potassium channels, intermediate/small-conductance calcium-activated potassium channels, eukaryotic single-domain voltage-gated cation channels, transient receptor potential channels, two-pore domain calcium channels, four-domain voltage-gated cation channels, cation and anion Cys-loop receptors, small-conductivity mechanosensitive channels, large-conductivity mechanosensitive channels, voltage-gated proton channels, inositole-1,4,5-trisphosphate receptors, slow anion channels, aluminum-activated malate transporters and quick anion channels, mitochondrial calcium uniporters, voltage-dependent anion channels, vesicular chloride channels, ionotropic purinergic receptors, animal volage-insensitive cation channels, channelrhodopsins, bestrophins, voltage-gated chloride channels H+/Cl- exchangers, plant calcium-permeable mechanosensitive channels, and trimeric intracellular cation channels. Overall, dinoflagellates represent cells able to respond to physical and chemical stimuli utilizing a wide range of G-protein coupled receptors- and Ca2+-dependent signaling pathways. The applied approach not only shed light on the ion channel set in dinoflagellates, but also provided the information on possible molecular mechanisms underlying vital cellular processes dependent on the ion transport.

MS-5, a Naphthalene Derivative, Induces the Apoptosis of an Ovarian Cancer Cell CAOV-3 by Interfering with the Reactive Oxygen Species Generation

  • Ma, Eunsook;Jeong, Seon-Ju;Choi, Joon-Seok;Nguyen, Thi Ha;Jeong, Chul-Ho;Joo, Sang Hoon
    • Biomolecules & Therapeutics
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    • v.27 no.1
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    • pp.48-53
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    • 2019
  • Reactive oxygen species (ROS) are widely generated in biological processes such as normal metabolism and response to xenobiotic exposure. While ROS can be beneficial or harmful to cells and tissues, generation of ROS by diverse anti-cancer drugs or phytochemicals plays an important role in the induction of apoptosis. We recently identified a derivative of naphthalene, MS-5, that induces apoptosis of an ovarian cell, CAOV-3. Interestingly, MS-5 induced apoptosis by down-regulating the ROS. Cell viability was evaluated by water-soluble tetrazolium salt (WST-1) assay. Apoptosis was evaluated by flow cytometry analysis. Intracellular ROS ($H_2O_2$), mitochondrial superoxide, mitochondrial membrane potential (MMP) and effect on cycle were determined by flow cytometry. Protein expression was assessed by western blotting. The level of ATP was measured using ATP Colorimetric/Fluorometric Assay kit. MS-5 inhibited growth of ovarian cancer cell lines, CAOV-3, in a concentration- and time-dependent manner. MS-5 also induced G1 cell cycle arrest in CAOV-3 cells, while MS-5 decreased intracellular ROS generation. In addition, cells treated with MS-5 showed the decrease in MMP and ATP production. In this study, we found that treatment with MS-5 in CAOV-3 cells induced apoptosis but decreased ROS level. We suspect that MS-5 might interfere with the minimum requirements of ROS for survival. These perturbations appear to be concentration-dependent, suggesting that MS-5 may induce apoptosis by interfering with ROS generation. We propose that MS-5 may be a potent therapeutic agent for inducing apoptosis in ovarian cancer cell through regulation of ROS.

Occurrence and control of N-nitrosodimethylamine in water engineering systems

  • Bian, Yongning;Wang, Chuang;Zhu, Guocheng;Ren, Bozhi;Zhang, Peng;Hursthouse, Andrew S.
    • Environmental Engineering Research
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    • v.24 no.1
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    • pp.1-16
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    • 2019
  • N-nitrosodimethylamine (NDMA) is a typical nitrogen disinfection by-product, which has posed a potential threat to human health during drinking water disinfection. Because of the well-known effects of mutagenesis, carcinogenesis and teratogenesis, the high detection rate in water engineering systems (such as coagulation, membrane filtration and biological systems), and difficulty to remove, it has received wide concern in the field of water engineering systems. The NDMA is a low molecular weight hydrophilic organic substance, which is difficult to remove. Also, the mechanism for NDMA formation is also recognized to be complex, and many steps still needed to be further evaluated. Therefore, the mechanistic knowledge on NDMA formation potential and their removal processes is of particularly interest. Few papers summarize the occurrence and control of NDMA in water engineering systems. It is for this reason that the content of this paper is particularly important for us to understand and control the amount of NDMA thus reducing the threat of disinfection by-products to drinking water. Four parts including the mechanisms for the NDMA formation potential, the factors affecting the NDMA formation potential, the technologies for removal of NDMA are summarized. Finally, some definite suggestions are given.

Evaluation of long-term water quality management policy effect using nonparametric statistical methods

  • Jung, Kang Young;Ahn, Jung Min;Cho, Sohyun;Lee, Yeong Jae;Han, Kun Yeun;Shin, Dongseok;Kim, Kyunghyun
    • Membrane and Water Treatment
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    • v.10 no.5
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    • pp.339-352
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    • 2019
  • Long term water quality change was analyzed to evaluate the effect of the Total Maximum Daily Load (TMDL) policy. A trend analysis was performed for biochemical oxygen demand (BOD) and total phosphorus (TP) concentrations data monitored at the outlets of the total 41 TMDL unit watersheds of the Nakdong River in the Republic of Korea. Because water quality data do not usually follow a normal distribution, a nonparametric statistical trend analysis method was used. The monthly mean values of BOD and TP for the period between 2004 and 2015 were analyzed by the seasonal Mann-Kendall test and the locally weighted scatterplot smoother (LOWESS). The TMDL policy effect on the water quality change of each unit watershed was analyzed together with the results of the trend analysis. From the seasonal Mann-Kendall test results, it was found that for BOD, 7.8 % of the 41 points showed downward trends, 26.8 % and the rest 65.9% showed upward and no trends. For TP, 51.2% showed no trends and the rest 48.8% showed downward trends. From the LOWESS analysis results, TP began to decrease in most of the unit watersheds from mid-2010s when intensive chemical treatment processes were introduced to existing wastewater treatment plants. Overall, for BOD, relatively more points were improved in the main stream compared to the points of the tributaries although overall trends were mostly no trend or upward. For TP, about half of the points were improved and the rest showed no trends.