• The Korean Journal of Physiology and Pharmacology
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• v.1 no.1
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• pp.97-105
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• 1997

The regulatory role of cyclic nucleotides in the expression of neutrophil responses has been examined. fMLP-stimulated superoxide production in neutrophils was inhibited by dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP), histamine, adenosine + theophylline, cAMP elevating agents, and 8-bromoguanosine 3' ,5' -cyclic monophosphate (8-BrcGMP) and sodium nitroprusside, cGMP elevating agents. Staurosporine, a protein kinase C inhibitor, genistein, a protein tyrosine kinase inhibitor and chlorpromazine, a calmodulin inhibitor, inhibited superoxide production by fMLP, but they did not further affect the action of DBcAMP on the stimulatory action of fMLP. DBcAMP, histamine, adenosine+theophylline and genistein inhibited myeloperoxidease release evoked by fMLP, whereas BrcGMP, sodium nitroprusside and staurosporine did not affect it. The elevation of $[Ca^{2+}]_i$ evoked by fMLP was inhibited by genistein and chlorpromazine but was not affected by staurosporine. DBcAMP exerted little effect on the initial peak in $[Ca^{2+}]_i$ response to fMLP but effectively inhibited the sustained rise. On the other hand, BrcGMP significantly inhibited both phases. fMLP-induced $Mn^{2+}$ influx was inhibited by either DBcAMP or BrcGMP. These results suggest that fMLP-stimulated neutrophil responses may be regulated by cAMP more than cGMP. cAMP and cGMP appear not affect stimulated responses by direct protein kinase C activation. Their regulatory action on the stimulated neutrophil responses may be not influenced by other activation processes.

• Tuberculosis and Respiratory Diseases
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• v.43 no.2
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• pp.210-220
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• 1996

Background : Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. In the process of surface adherence, adhesion molecules have a clear role in intracellular signal pathway of cellular activation. Human alveolar macrophages(HAM) are frequently purified by the adherence procedure after bronchoalveolar lavage. But the experimental data of many reports about alveolar macrophages have ignored the possibility of adhesion-induced cellular activation. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be normal by chest CT. With the measurement of hydrogen peroxide release from adherent HAM to plastic surface and non-adherent HAM with or without additional stimulation of phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP), we observed the effect of the adherence to plastic surface. We also evaluated the effect of various biological surfaces on adhesion-induced activation of HAM. Then, to define the intracellular pathway of signal transduction, pretreatment with cycloheximide, pertussis toxin and anti-CD11/CD18 monoclonal antibody was done and we measured hydrogen peroxide in the culture supernatant of HAM. Results : 1) The adherence itself to plastic surface directly stimulated hydrogen peroxide release from human alveolar macrophages and chemical stimuli such as phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine(fMLP) colud not increase hydrogen peroxide release in these adherent macrophages which is already activated. 2) PMA activated human alveolar macrophages irrespective of the state of adhesion. However, fMLP stimulated the release of hydrogen peroxide from the adherent macrophages, but not from the non-adherent macrophages. 3) HAM adherent to A549 cell(type II alveolar epithelium-like human cell line) monolayer released more hydrogen peroxide in response to both PMA and fMLP. This adherence-dependent effect of fMLP was blocked by pretreatment of macrophages with cycloheximide, pertussis toxin and anti-CD18 monoclonal antibody, Conclusion : These results suggest that the stimulatory effect of PMA and fMLP can not be found in adherent macrophage because of the activation of human alveolar macrophage by the adherence to plastic surface and the cells adhered to biologic surface such as alveolar epithelial cells are appropriately responsive to these stimuli. It is also likely that the effect of fMLP on the adherent macrophage requires new protein synthesis via G protein pathway and is dependent on the adhesion between alveolar macrophages and alveolar epithelial cells by virtue of CD11/CD18 adhesion molecules.

• The Korean Journal of Pharmacology
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• v.31 no.3
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• pp.333-344
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• 1995

The effects of adenosine and $N^6-cyclopentyladenosine$ (CPA) on superoxide production, myeloperoxidase release and $Ca^{2+}$ mobilization stimulated by fMLP in neutrophils were investigated. The effects were also observed on the stimulatory actions of C5a and PMA and the responses in lipopolysaccharide-primed neutrophils. In addition, the involvement of cAMP in the inhibitory action of adenosine was examined. The fMLP-stimulated neutrophil respiratory burst, degranulation and intracellular $Ca^{2+}$ mobilization may be regulated by activation of adenosine receptors. Adenosine may not affect the stimulated neutrophil responses due to activation of protein kinase C. fMLP-stimulated respiratory burst in lipopolysaccharide-primed neutrophils may be less sensitive to adenosine, compared with nonprimed cells. The inhibitory effect of theophylline in the presence of adenosine on neutrophil responses appears to be ascribed to accumulation of intracellular cAMP.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.6
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• pp.497-502
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• 2014

Extracellular nicotinamide adenine dinucleotide (NAD) cleaving activity of a particular cell type determines the rate of the degradation of extracellular NAD with formation of metabolites in the vicinity of the plasma membrane, which has important physiological consequences. It is yet to be elucidated whether intact human neutrophils have any extracellular NAD cleaving activity. In this study, with a simple fluorometric assay utilizing $1,N^6$-ethenoadenine dinucleotide (etheno-NAD) as the substrate, we have shown that intact peripheral human neutrophils have scant extracellular etheno-NAD cleaving activity, which is much less than that of mouse bone marrow neutrophils, mouse peripheral neutrophils, human monocytes and lymphocytes. With high performance liquid chromatography (HPLC), we have identified that ADP-ribose (ADPR) is the major extracellular metabolite of NAD degradation by intact human neutrophils. The scant extracellular etheno-NAD cleaving activity is decreased further by N-formyl-methionine-leucine-phenylalanine (fMLP), a chemoattractant for neutrophils. The fMLP-mediated decrease in the extracellular etheno-NAD cleaving activity is reversed by WRW4, a potent FPRL1 antagonist. These findings show that a much less extracellular etheno-NAD cleaving activity of intact human neutrophils compared to other immune cell types is down-regulated by fMLP via a low affinity fMLP receptor FPRL1.

• Journal of Life Science
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• v.13 no.6
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• pp.903-910
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• 2003

Phospholipase D (PLD) plays an important role as a signaling molecule in the activation of neutrophils. In this study, effect of nitric oxide (NO) and cGMP on the activation of PLD in human neutrophils was investigated. Sodium nitroprusside (SNP), an agent to produce NO spontaneously in cells, alone increased PLD activity and the maximal activation was obtained with 0.5 mM SNP. Dibutyryl-cAMP, an agent to increase an intracellular cAMP concentration inhibited formyl-Met-Leu-Phe (fMLP)-stimulated PLD activity but 8-bromo-cGMP (300 $\mu$M), an agent to increase an intracellular cGMP concentration did not affect basal and fMLP-stimulated PLD activity. NO-induced activation of PLD was not blocked by KT 5823, an inhibitor of cGMP-dependent protein kinase (PKG), suggesting that NO-induced PLD activation is not mediated by cGMP. NO also stimulated p38 mitogen activated protein kinase (MAPK) in human neutrophils, indicated by increased phosphorylation of p38 MAPK in Western blotting. NO-induced phosphorylation of p38 MAPK was not inhibited by KT 5823 or n-butanol. RhoA, an regulatory factor of PLD activation was trans-located from cytosolic fraction to plasma membranes by fMLP or phorbol ester, and fMLP-stimulated but not phorbol ester-stimulated translocation of RhoA was inhibited by cGMP. These results suggest that NO stimulates PLD activity through other unidentified facto.(s) than cGMP even though cGMP inhibits the artivation of RhoA.

• Journal of Korea Water Resources Association
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• v.49 no.10
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• pp.877-885
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• 2016

In this study, the Standard Precipitation Index(SPI), meteorological drought index, was used to evaluate the temporal and spatial assessment of drought forecasting results for all cross Korea. For the drought forecasting, the Multi Layer Perceptron-Artificial Neural Network (MLP-ANN) was selected and the drought forecasting was performed according to different forecasting lead time for SPI (3) and SPI (6). The precipitation data observed in 59 gaging stations of Korea Meteorological Adminstration (KMA) from 1976~2015. For the performance evaluation of the drought forecasting, the binary classification confusion matrix, such as evaluating the status of drought occurrence based on threshold, was constituted. Then Receiver Operating Characteristics (ROC) score and F score according to conditional probability are computed. As a result of ROC analysis on forecasting performance, drought forecasting performance, of applying the MLP-ANN model, shows satisfactory forecasting results. Consequently, two-month and five-month leading forecasts were possible for SPI (3) and SPI (6), respectively.

• The Korean Journal of Mycology
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• v.15 no.1
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• pp.1-8
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• 1987

The purpose of this experiment is to examine if G. lucidum extracts could have an effect on leukocyte migration and to select the best chemotactic factor for leukocyte migration. When chemotactic factors such as TC-199 medium, filtrate of E. coli culture, N-fMLP, and G. lucidum extracts were used, N-fMLP, G. lucidum extracts and filtrate of E. coli were positive effect, but TC-199 medium was less than others. The various concentrations of the chemotactic factors such as G. lucidum extracts(1.00%, 0.10% and 0.01%) were stimulated for the leukocytes migration in the modified Boyden chambers, however, the lowest concentration such as 0.01% was more effective than others for the chemoattractant of the leukocyte migration When the leukocyte was treated with G. lucidum extracts(1.00%, 0.10%, and 0.01%) at room temperature for 120 minutes, the chemotactic activities had a good effect at 0.01%. Therefore, leukocyte was affected by G. lucidum extracts.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.1031-1037
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• 2004

The interaction of chemoattractant receptors and $G{\alpha}_{16}$ was studied to provide the molecular basis to elucidate the interaction of chemoattractant receptors with $G{\alpha}_{16}$ subunit, thereby possibly contributing to finding novel targets for designing new type of G protein antagonists with anti-inflammatory effects. Experiments were performed to characterize the $G{\alpha}_{16}$ subunit domains responsible for efficient coupling to chemoattractant receptors. Thus, a series of chimeric $G{\alpha}_{11}G{\alpha}_{16}$ and $G{\alpha}_{16}G{\alpha}_{11}$ cDNA constructs were expressed, and the ability of chimeric proteins to mediate C5a, IL-8, and fMLP-induced release of inositol phosphate in transfected Cos-7 cells was tested. The results showed that short stretches of residues 154 to residue 167 and from residue 174 to residue 195 of $G{\alpha}_{16}$ contribute to efficient coupling to the C5a receptor. On the other hand, a stretch of amino acid residues 220-240 of $G{\alpha}_{16}$ that is necessary for interacting with C5a receptor did not play any role in the interaction with IL-8 receptor. However, a stretch from residue 155 to residue 195 of $G{\alpha}_{16}$ was found to be crucial for efficient coupling to IL-8 receptor in concert with C-terminal 30 amino acid residues of this ${\alpha}$ subunit. Coupling profiles of a variety of chimeras, composed of $G{\alpha}_{11}G{\alpha}_{16}$ to fMLP receptor indicate that the C-terminal 30 amino acids are most critical for the coupling of $G{\alpha}_{16}$ to fMLP receptor. Taken together, $G{\alpha}_{16}$ subunit recruits multiple and distinctive coupling regions, depending on the type of receptors, to interact.

• Journal of Internet Computing and Services
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• v.12 no.6
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• pp.63-71
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• 2011

Cardiac disorder data are generally tested using the classifier and QRS-Complex and R-R interval which is used in this experiment are often extracted by ECG(Electrocardiogram) signals. The experimentation of ECG data with classifier is generally performed with SVM(Support Vector Machine) and MLP(Multilayer Perceptron) classifier, but this study experimented with Best-First Decision Tree(B-F Tree) derived from the Dicision Tree among Random Forest classifier algorithms to improve accuracy. To compare and analyze accuracy, experimentation of SVM, MLP, RBF(Radial Basic Function) Network and Decision Tree classifiers are performed and also compared the result of announced papers carried out under same interval and data. Comparing the accuracy of Random Forest classifier with above four ones, Random Forest is the best in accuracy. As though R-R interval was extracted using Band-pass filter in pre-processing of this experiment, in future, more filter study is needed to extract accurate interval.

• Journal of the Korea Convergence Society
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• v.10 no.10
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• pp.7-13
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• 2019

The natural gas (NG), mostly methane leaks into the air, it is a big problem for the climate. detected NG leaks under U.S. city streets and collected data. In this paper, we introduced a Deep Neural Network (DNN) classification of prediction for a level of NS leak. The proposed method is OrdinalEncoder(OE) based K-means clustering and Multilayer Perceptron(MLP) for predicting NG leak. The 15 features are the input neurons and the using backpropagation. In this paper, we propose the OE method for labeling target data using k-means clustering and compared normalization methods performance for NG leak prediction. There five normalization methods used. We have shown that our proposed OE based MLP method is accuracy 97.7%, F1-score 96.4%, which is relatively higher than the other methods. The system has implemented SPSS and Python, including its performance, is tested on real open data.