• Title/Summary/Keyword: Generation of Mechanisms

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Effects of Noradrenaline on the Spontaneous Contraction and Ionic Current in the Antral Circular Muscle of Guinea-pig Stomach

  • Jun, Jae-Yeoul;Lee, Sang-Jin;Kim, Sung-Joon;Suh, Jae-Yul;So, In-Suk;Hwang, Sang-Ik;Kim, Ki-Whan
    • The Korean Journal of Physiology
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    • v.27 no.2
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    • pp.115-122
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    • 1993
  • There is evidence that noradrenaline enhances spontaneous contractions dose-dependently in guinea-pig antral circular muscle. To investigate the mechanism of this excitatory action, slow waves and membrane currents were recorded using conventional microelectrode techniques in muscle strips and the whole cell patch clamp technique in isolated gastric myocytes. On recording slow waves, noradrenaline $(10^{-5}\;M)$ induced the hyperpolarization of the membrane potential, although the shape of the slow waves became tall and steep. Also, spike potentiaIs occurred at the peaks of slow waves. These changes were completely reversed by administration of phentolamine $(10^{-5}\;M),\;an\;{\alpha}-adrenoceptor$ blocker. Noradrenaline-induced hyperpolarization was blocked by apamin $(10^{-7}\;M)$, a blocker of a class of $Ca^{2+}\;-dependent\;K^+$ channels. To investigate the mechanisms for these effects, we performed whole cell patch clamp experiments. Norndrenaline increased voltage-dependent $Ca^{2+}$ currents in the whole range of test potentials. Noradrenaline also increased $Ca^{2+}\;-dependent\;K^+$\;currents, and this effects was abolished by apamin. These results suggest that the increase in amplitude and the generation of spike potentials on slow waves was caused by the activation of voltage-dependent $Ca^{2+}$ channel via adrenoceptors, and hyperpolarization of the membrane potential was mediated by activation of apamin-sensitive $Ca^{2+}\;-dependent\;K^+\;channels$.

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Developing New Mammalian Gene Expression Systems Using the Infectious cDNA Molecular Clone of the Japanese Encephalitis Virus

  • Yun Sang-Im;Choi Yu-Jeong;Park Jun-Sun;Kim Seok-Yong;Lee Young-Min
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2003.05a
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    • pp.83-86
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    • 2003
  • Major advances in positive-sense RNA virus research have been facilitated by the development of reverse genetics systems. These systems consist of an infectious cDNA clone that encompasses the genome of the virus in question. This clone is then used as a template for the subsequent synthesis of infectious RNA for the generation of synthetic viruses. However, the construction of infectious cDNA for the Japanese encephalitis virus (JEV) has been repeatedly thwarted by the instability of its cDNA. As JEV is an important human pathogen that causes permanent neuropsychiatric sequelae and even fatal disease, a reliable reverse genetics system for this virus is highly desirable. The availability of this tool would greatly and the development of effective vaccines as well as facilitate studies into the basic biology of the virus, including the molecular mechanisms of viral replication, neurovirulence, and pathogenesis. We have successfully constructed a genetically stable infectious JEV cDNA containing full-length viral RNA genome. Synthetic RNA transcripts generated in vitro from the cDNA were highly infectious upon transfection into susceptible cells, and the cDNA remained stable after it had been propagated in E. coli for 180 generations. Using this infectious JEV cDNA, we have successfully expressed a variety of reporter genes from the full-length genomic and various subgenomic RNAs in vitro transcribed from functional JEV cDNAS. In summary, we have developed a reverse genetics system for JEV that will greatly facilitate the research on this virus in a variety of different fields. It will also be useful as a heterologous gene expression vector and aid the development of a vaccine against JEV.

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Proteomic Analysis of the Aging-related Proteins in Human Normal Colon Epithelial Tissue

  • Li, Ming;Xiao, Zhi-Qiang;Chen, Zhu-Chu;Li, Jian-Ling;Li, Cui;Zhang, Peng-Fei;Li, Mao-Yu
    • BMB Reports
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    • v.40 no.1
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    • pp.72-81
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    • 2007
  • In order to screen the aging related proteins in human normal colon epithelia, the comparative proteomics analysis was applied to get the two-dimensional electrophoresis (2-DE) profiles with high resolution and reproducibility from normal colon epithelial tissues of young and aged people. Differential proteins between the colon epithelia of two age groups were found with PDQuest software. The thirty five differential protein-spots were identified by peptide mass fingerprint (PMF) based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and database searching. Among them there are sixteen proteins which are significantly up-regulated in the colonic mucosal epithelia of young people group, which include ATP synthase beta chain, electron transfer flavoprotein alpha-subunit, catalase, glutathione peroxidase 1, annexin A2 and heat shock cognate 71 kDa protein, etc.; There are nineteen proteins which are significantly up-regulated in the colonic mucosal epithelia of aged people group, which include far upstream element-binding protein 1, nucleoside diphosphate kinase B, protein disulfide-isomerase precursor and VDAC-2, etc.. The identified differential proteins appear to be involved in metabolism, energy generation, chaperone, antioxidation, signal transduction, protein folding and apoptosis. The data will help to understand the molecular mechanisms of human colon epithelial aging.

Chongmyung-tang Inhibits the Cytotoxicity of Beta-amyloid in Neuro 2A Neuroblastoma Cells (베타 아밀로이드 유도성 Neuro 2A 세포독성에 대한 총명탕의 효과)

  • Gug Yun Jai;Choi Hyuk;Kim Tae Heon;Kang Hyung Won;Lyu Young Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1418-1425
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    • 2004
  • The water extract of Chongmyung-tang has been traditionally used for treatment of memory-disorder in oriental medicine. This study was designed to investigate the protective mechanisms of Chongmyung-tang on β-amyloid or H₂O₂-induced cytotoxicity in Neuro 2A cells. The water extract of Chongmyung-tang significantly reduced both β-amyloid or H₂O₂-induced cell death and apoptotic characteristics through reduction of intracellular peroxide generation. Also, it inhibited the mitochondrial dysfunction including the disruption of mitochondria membrane permeability transition(MPT) and the modulation in expression of Bcl-2 family proteins in H₂O₂-treated H9c2 cells. Furthermore, pretreatment of quercetin inhibited the activation of caspase-3, in turn, degradation of ICAD/DFF45 were completely abolished in H₂O₂-treated cells. Taken together, that data suggest that the protective effects of the water extract of Chongmyung-tang against β-amyloid induced oxidative injuries may be achieved through modulation of mitochondrial dysfunction.

Reduction of TNE ${\alpha}-induced$ Oxidative DNA Damage Product, 8-Hydroxy-2'-Deoxyguanosine, in L929 Cells Stably Transfected with Small Heat Shock Protein

  • Park, Young-Mee;Choi, Eun-Mi
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.2
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    • pp.209-219
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    • 1997
  • Previous studies have demonstrated that oxidative stress involving generation of reactive oxygen species (ROS) is responsible for the cytotoxic action of $TNF{\alpha}$. Protective effect of small heat shock proteins (small HSP) against diverse oxidative stress conditions has been suggeted. Although overexpression of small hsp was shown to provide an enhanced survival of $TNF{\alpha}$-sensitive cells when challenged with $TNF{\alpha}$, neither the nature of $TNF{\alpha}$-induced cytotoxicity nor the protective mechanism of small HSP has not been completely understood. In this study, we have attempted to determine whether $TNF{\alpha}$ induces oxidative DNA damage in $TNF{\alpha}$-sensitive L929 cells. We chose to measure the level of 8-hydroxy-2'-deoxyguanosine (8 ohdG), which has been increasingly recognized as one of the most sensitive markers of oxidative DNA damage. Our results clearly demonstrated that the level of 8 ohdG increased in L929 cells in a $TNF{\alpha}$ dose-dependent manner. Subsequently, we asked whether small HSP has a protective effect on $TNF{\alpha}$-induced oxidative DNA damage. To accomplish this goal, we have stably transfected L929 cells with mouse small hsp cDNA (hsp25) since these cells are devoid of endogenous small hsps. We found that $TNF{\alpha}$-induced 8 ohdG was decreased in cells overexpressing exogenous small hsp. We also found that the cell killing activity of $TNF{\alpha}$ was decreased in these cells as measured by clonogenic survival. Taken together, results from the current study show that cytotoxic mechanism of $TNF{\alpha}$ involves oxidative damage of DNA and that overexpression of the small hsp reduces this oxidative damage. We suggest that the reduction of oxidative DNA damage is one of the most important protective mechanisms of small HSP against $TNF{\alpha}$.

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γ-Aminobutyric acid (GABA) confers chromium stress tolerance in mustard (Brassica juncea L.) seedlings by modulating the antioxidant defense and glyoxalase systems

  • Al Mahmud, Jubayer;Hasanuzzaman, Mirza;Nahar, Kamrun;Rahman, Anisur;Hossain, Md. Shahadat;Fujita, Masayuki
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.235-235
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    • 2017
  • Chromium (Cr) toxicity is hazardous to the seed germination, growth, and development of plants. ${\gamma}$-Aminobutyric acid (GABA) is a non-protein amino acid and is involved in stress tolerance in plants. To investigate the effects of GABA in alleviating Cr toxicity, we treated eight-d-old mustard (Brassica juncea L.) seedlings with Cr (0.15 mM and 0.3 mM $K_2CrO_4$, 5 days) alone and in combination with GABA ($125{\mu}M$) in a semi-hydroponic medium. The roots and shoots of the seedlings accumulated Cr in a dose-dependent manner, which led to an increase in oxidative damage [lipid peroxidation; hydrogen peroxide ($H_2O_2$) content; superoxide ($O{_2}^{{\cdot}-}$) generation; lipoxygenase (LOX) activity], MG content, and disrupted antioxidant defense and glyoxalase systems. Chromium stress also reduced growth, leaf relative water content (RWC), and chlorophyll (chl) content but increased phytochelatin (PC) and proline (Pro) content. Furthermore, supplementing the Cr-treated seedlings with GABA reduced Cr uptake and upregulated the non-enzymatic antioxidants (ascorbate, AsA; glutathione, GSH) and the activities of the enzymatic antioxidants including ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX), superoxide dismutase (SOD), catalase (CAT), glyoxalase I (Gly I), and glyoxalase II (Gly II), and finally reduced oxidative damage. Adding GABA also increased leaf RWC and chl content, decreased Pro and PC content, and restored plant growth. These findings shed light on the effect of GABA in improving the physiological mechanisms of mustard seedlings in response to Cr stress.

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Anti-neuroinflammatory effects of ethanolic extract of black chokeberry (Aronia melanocapa L.) in lipopolysaccharide-stimulated BV2 cells and ICR mice

  • Lee, Kang Pa;Choi, Nan Hee;Kim, Hyun-Soo;Ahn, Sanghyun;Park, In-Sik;Lee, Dea Won
    • Nutrition Research and Practice
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    • v.12 no.1
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    • pp.13-19
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    • 2018
  • BACKGROUND/OBJECTIVES: One of the mechanisms considered to be prevalent in the development of Alzheimer's disease (AD) is hyper-stimulation of microglia. Black chokeberry (Aronia melanocapa L.) is widely used to treat diabetes and atherosclerosis, and is known to exert anti-oxidant and anti-inflammatory effects; however, its neuroprotective effects have not been elucidated thus far. MATERIALS/METHODS: We undertook to assess the anti-inflammatory effect of the ethanolic extract of black chokeberry friut (BCE) in BV2 cells, and evaluate its neuroprotective effect in the lipopolysaccharide (LPS)-induced mouse model of AD. RESULTS: Following stimulation of BV2 cells by LPS, exposure to BCE significantly reduced the generation of nitric oxide as well as mRNA levels of numerous inflammatory factors such as inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), interleukin 1 beta ($IL-1{\beta}$), and tumor necrosis factor alpha ($TNF-{\alpha}$). In addition, AD was induced in a mouse model by intraperitoneal injection of LPS ($250{\mu}g/kg$), subsequent to which we investigated the neuroprotective effects of BCE (50 mg/kg) on brain damage. We observed that BCE significantly reduced tissue damage in the hippocampus by downregulating iNOS, COX-2, and $TNF-{\alpha}$ levels. We further identified the quinic acids in BCE using liquid chromatography-mass spectrometry (LCMS). Furthermore, we confirmed the neuroprotective effect of BCE and quinic acid on amyloid beta-induced cell death in rat hippocampal primary neurons. CONCLUSIONS: Our findings suggest that black chokeberry has protective effects against the development of AD.

Protective Effect of Radix Clematidis Extract on Streptozotocin-induced Diabetes (Streptozotocin 유도 당뇨병에 대한 위령선(威靈仙) 추출물의 방어 효과)

  • Ham, Kyung-Wan;Kim, Eun-Kyung;Song, Mi-Young;Kwon, Kang-Beom;Song, Je-Ho;Seo, Eun-A;Ryu, Do-Gon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.3
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    • pp.580-584
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    • 2008
  • In the present study, Radix clematidis extract (RCE) was evaluated to determine if it could protect pancreatic ${\beta}$ cells against multiple low dose streptozotocin (MLDS)-induced diabetes. Injection of mice with MLDS resulted in hyperglycemia and hypoinsulinemia, which was confirmed by immunohistochemical staining. However, the induction of diabetes by MLDS was completely prevented when mice were pre-administrated with RCE. Generation of oxidative stress is implicated in MLDS, a ${\beta}$ cell specific toxin-induced islet cell death. In this context, to elucidate the mechanisms of protective effects in RCE pre-administrated diabetic mice, we investigated the expression of heme oxygenase-1 (HO-1), which is one of the anti-oxidant enzymes. MLDS-induced HO-1 expressions were significantly reduced in MLDS-treated mice. However, the decrease of HO-1 by MLDS were protected by pretreatment of RCE. The molecular mechanism by which RCE inhibits diabetic conditions by MLDS appears to involve inhibition of HO-1 expression. Taken together, these results reveal the possible therapeutic value of RCE for the prevention of type 1 diabetes progression.

Danchunhwan Protects the Cytotoxicity of Beta-amyloid in SH-SY5Y Neuroblastoma Cells (베타아밀로이드 유도성 SH-SY5Y 세포독성에서 단천환(丹川丸)의 보호효과)

  • Yu, Bong-Sun;Kim, Jin-Kyung;;Park, Chan-Ny;So, Hong-Seob
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.6
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    • pp.1516-1523
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    • 2006
  • The water extract of Danchunhwan(DCH) has been traditionally used for treatment of dementia damage in oriental medicine. However, little is known about the mechanism by which the water extract of DCH rescues cells from neurodegenerative disease such as Alzheimer's disease. This study was designed to investigate the protective mechanisms of DCH on ${\beta}$-amyloid or $H_2O_2$-induced cytotoxicity in SH-SY5Y neuronblastoma cells. ${\beta}$-amyloid and $H_2O_2$ markedly decreased the viability of SH-SY5Y cells, which was characterized with apparent apoptotic features such as membrane blebbing as well as fragmentation of genomic DNA and nuclei. However, the water extract of DCH significantly reduced both ${\beta}$-amyloid or $H_2O_2$-induced cell death and apoptotic characteristics through reduction of intracellular peroxide generation. Also, the water extract of DCH prevented prevented the mitochondrial dysfunction including the disruption of mitochondria membrane permeability transition (MPT) and the perturbation in Bcl-2 family protein expressions in $H_2O_2$-treated SH-SY5Y cells.

Characteristics of EMR emitted by coal and rock with prefabricated cracks under uniaxial compression

  • Song, Dazhao;You, Qiuju;Wang, Enyuan;Song, Xiaoyan;Li, Zhonghui;Qiu, Liming;Wang, Sida
    • Geomechanics and Engineering
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    • v.19 no.1
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    • pp.49-60
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    • 2019
  • Crack instability propagation during coal and rock mass failure is the main reason for electromagnetic radiation (EMR) generation. However, original cracks on coal and rock mass are hard to study, making it complex to reveal EMR laws and mechanisms. In this paper, we prefabricated cracks of different inclinations in coal and rock samples as the analogues of the native cracks, carried out uniaxial compression experiments using these coal and rock samples, explored, the effects of the prefabricated cracks on EMR laws, and verified these laws by measuring the surface potential signals. The results show that prefabricated cracks are the main factor leading to the failure of coal and rock samples. When the inclination between the prefabricated crack and axial stress is smaller, the wing cracks occur first from the two tips of the prefabricated crack and expand to shear cracks or coplanar secondary cracks whose advance directions are coplanar or nearly coplanar with the prefabricated crack's direction. The sample failure is mainly due to the composited tensile and shear destructions of the wing cracks. When the inclination becomes bigger, the wing cracks appear at the early stage, extend to the direction of the maximum principal stress, and eventually run through both ends of the sample, resulting in the sample's tensile failure. The effect of prefabricated cracks of different inclinations on electromagnetic (EM) signals is different. For samples with prefabricated cracks of smaller inclination, EMR is mainly generated due to the variable motion of free charges generated due to crushing, friction, and slippage between the crack walls. For samples with larger inclination, EMR is generated due to friction and slippage in between the crack walls as well as the charge separation caused by tensile extension at the cracks' tips before sample failure. These conclusions are further verified by the surface potential distribution during the loading process.