• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.219-227
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• 2008

The free-living photoheterotrophic Gram-negative bacterium Rhodobacter sphaeroides possesses a quorum-sensing (QS) regulatory system mediated by CerR-CerI, a member of the LuxR-LuxI family. To identify the genes affected by the regulatory system, random lacZ fusions were generated in the genome of R. sphaeroides strain 2.4.1 using a promoter-trapping vector, pSG2. About 20,000 clones were screened and 23 showed a significantly different level of ${\beta}$-gal activities upon the addition of synthetic 7,8-cis-N-tetradecenoyl-homoserine lactone (RAI). Among these 23 clones, the clone showing the highest level of induction was selected for further study, where about a ten-fold increase of ${\beta}$-gal activity was exhibited in the presence of RAI and induction was shown to be required for cerR. In this clone, the lacZ reporter was inserted in a putative gene that exhibited a low homology with catD. A genetic analysis showed that the expression of the catD homolog was initiated from a promoter of another gene present upstream of the catD. This upstream gene showed a strong homology with luxR and hence was named qsrR (quorum-sensing regulation regulator). A comparison of the total protein expression profiles for the wild-type cells and qsrR-null mutant cells using two-dimensional gel electrophoresis and a MALDI-TOF analysis allowed the identification of sets of genes modulated by the luxR homolog.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.3
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• pp.275-282
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• 1999

Muscle strips and muscle cells from cat stomach were used to investigate whether spontaneously formed cyclic nucleotides were involved in the inhibition of gastric smooth muscle contraction. A phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IBMX), increased the levels of both cyclic GMP (cGMP) and cyclic AMP (cAMP) in resting state cells, while decreasing acetylcholine-induced muscle contraction. Under the influence of IBMX, SQ22536, an adenylyl cyclase inhibitor and methylene blue, a guanylyl cyclase inhibitor completely blocked increases in cAMP and cGMP respectively, without any effect on contraction. However, the combination of SQ22536 and methylene blue completely blocked increases in both cAMP and cGMP levels and stimulated contractions markedly even in the presence of IBMX. Muscle contraction inhibitors such as isoprenaline, vasoactive intestinal polypeptide and sodium nitroprusside also appeared to increase cyclic nucleotide levels which decreased contraction. Which nucleotide increased the most was dependent on the agonist used. Therefore, irrespective of the cyclic nucleotide class, the spontaneous formation of cyclic nucleotides should be considered in evaluating the mechanism of gastric smooth muscle relaxation.

• Archives of Pharmacal Research
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• v.29 no.5
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• pp.400-404
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• 2006

This study examined the effect of Gamma-Amino butyric acid (GABA) and selective GABA receptor related drugs on the electrically stimulated relaxation in the lower esophageal sphincter muscle (LES) of a cat. Tetrodotoxin $(10^{-6}\;M)$ suppressed the electrically stimulated (0.5-5 Hz) relaxation of the LES. However, guanethidine $(10^{-6}\;M)$ and atropine $(10^{-6}\;M)$ had no effect indicating that the relaxations were neurally mediated via the nonadrenergic and noncholinergic (NANC) pathways. NG-nitro-L-arginine methyl ester ($10^{-4}M$, L-NAME) also inhibited the relaxant response but did not completely abolish the electrically stimulated relaxation with 60% inhibition, which suggests the involvement of nitric oxide as an inhibitory transmitter. This study examined the role of GABA, an inhibitory neurotransmitter, on neurally mediated LES relaxation. GABA ($10^{-3}-10^{-5}M$, non selective receptor agonist), muscimol ($10^{-3}-10^{-5}M$, GABA-A agonist), and baclofen ($10^{-3}-10^{-5}M$, GABA-B agonist) had no significant effect on the electrically stimulated relaxation. Moreover, bicuculline ($10^{-5}M$, GABA-A antagonist) and phaclofen ($10^{-5}M$, GABA-B antagonist) had no inhibitory effect on the electrically stimulated relaxation. This suggests that GABA and the GABA receptor are not involved in the electrically stimulated NANC relaxation in the cat LES.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.417-425
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• 1995

The regional distribution and relative frequencies of gastrointestinal endocrine cells were studied immunohistochemically in the GIT of the cat-shark. Five kinds of endocrine cells were identified in this study. Mostly these cells were of open types, except for glucagon-, BPP- and somatostatin-immunoreactive cells in the stomach which seemed to be of closed type. 5-HT-immunoreactive cells were detected throughout the GIT, and were more freguent than of the other regions. Glucagon-and BPP-immunoreactive cells were demonstrated in the entire GIT except for the duodenum an the rectum, and occurred in various frequencies. A few numbers of Gas/CCK-immunoreactive cells were restricted to the duodenum. Somatostatin-immunoreactive cells were distributed in the whole GIT, and were a few in numbers in the stomach and rare in the intestine, respectly. These results suggest that the pattern of the regional distribution is rather similar to that reported fur previously, but relative number was less frequent than that of other species.

• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.577-586
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• 2019

The engineered Aspergillus oryzae has a high NADPH demand for xylose utilization and overproduction of target metabolites. Glucose-6-phosphate dehydrogenase (G6PDH, E.C. 1.1.1.49) is one of two key enzymes in the oxidative part of the pentose phosphate pathway, and is also the main enzyme involved in NADPH regeneration. The open reading frame and cDNA of the putative A. oryzae G6PDH (AoG6PDH) were obtained, followed by heterogeneous expression in Escherichia coli and purification as a his6-tagged protein. The purified protein was characterized to be in possession of G6PDH activity with a molecular mass of 118.0 kDa. The enzyme displayed maximal activity at pH 7.5 and the optimal temperature was $50^{\circ}C$. This enzyme also had a half-life of 33.3 min at $40^{\circ}C$. Kinetics assay showed that AoG6PDH was strictly dependent on $NADP^+$ ($K_m=6.3{\mu}M$, $k_{cat}=1000.0s^{-1}$, $k_{cat}/K_m=158.7s^{-1}{\cdot}{\mu}M^{-1}$) as cofactor. The $K_m$ and $k_{cat}/K_m$ values of glucose-6-phosphate were $109.7s^{-1}{\cdot}{\mu}M^{-1}$ and $9.1s^{-1}{\cdot}{\mu}M^{-1}$ respectively. Initial velocity and product inhibition analyses indicated the catalytic reaction followed a two-substrate, steady-state, ordered BiBi mechanism, where $NADP^+$ was the first substrate bound to the enzyme and NADPH was the second product released from the catalytic complex. The established kinetic model could be applied in further regulation of the pentose phosphate pathway and NADPH regeneration of A. oryzae to improve its xylose utilization and yields of valued metabolites.

• Steel and Composite Structures
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• v.51 no.6
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• pp.679-695
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• 2024

This paper presents a novel technique that combines machine learning (ML) with moth-flame optimization (MFO) methods to predict the axial compressive strength (ACS) of concrete filled double skin steel tubes (CFDST) columns. The proposed model is trained and tested with a dataset containing 125 tests of the CFDST column subjected to compressive loading. Five ML models, including extreme gradient boosting (XGBoost), gradient tree boosting (GBT), categorical gradient boosting (CAT), support vector machines (SVM), and decision tree (DT) algorithms, are utilized in this work. The MFO algorithm is applied to find optimal hyperparameters of these ML models and to determine the most effective model in predicting the ACS of CFDST columns. Predictive results given by some performance metrics reveal that the MFO-CAT model provides superior accuracy compared to other considered models. The accuracy of the MFO-CAT model is validated by comparing its predictive results with existing design codes and formulae. Moreover, the significance and contribution of each feature in the dataset are examined by employing the SHapley Additive exPlanations (SHAP) method. A comprehensive uncertainty quantification on probabilistic characteristics of the ACS of CFDST columns is conducted for the first time to examine the models' responses to variations of input variables in the stochastic environments. Finally, a web-based application is developed to predict ACS of the CFDST column, enabling rapid practical utilization without requesting any programing or machine learning expertise.

• Bulletin of the Korean Chemical Society
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• v.34 no.9
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• pp.2671-2674
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• 2013

To elucidate the role of the evolutionarily conserved Tyr8 residue in rice Phi-class GSTF3, this amino acid was replaced with alanine and phenylalanine by site-directed mutagenesis, respectively. The replacement of Tyr8 with Ala significantly affected the catalytic activity and the kinetic parameters, whereas the substitutions of Tyr8 with Phe had almost no effect. The Y8A mutant resulted in approximately 90-100% decrease of the specific activity. Moreover, the Y8A mutant resulted approximately in 2-fold increase of $K_m$, approximately 60-80% decrease of $k_{cat}$, and approximately 6.5-fold decrease in $k_{cat}/K_m$. From the pH/log $k_{cat}/K_m$ plot, $pK_a$ values of the GSH in the wild-type enzyme-GSH complex, Y8A-GSH complex and Y8F-GSH complex were estimated to be approximately 6.8, 8.5 and 6.9, respectively. From these results, we suggest that the evolutionarily conserved Tyr8 residue in OsGSTF3 seems to influence the structural stability of the active site of OsGSTF3 rather than directly its catalytic activity.

• Journal of Food Hygiene and Safety
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• v.26 no.4
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• pp.370-376
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• 2011

This study was performed to investigate the antioxidative effect of Ligusticum chuanxiong Hort extracts (LCE) against the hyperlipidemia of high-fat diet-fed obese rats. The rats were divided into the three groups (normal group, control group and sample group) to perform the experimental research. 1.5 ml/kg of LCE was intraperitoneally administered into the sample group for 21 days. The equal dose of 0.9% saline was intraperitoneally administered into the normal group and the control group. On day 22, they were anesthetized with ether and dissected. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) were examined in serum of rats. Superoxide dismutase (SOD) was measured in mitochondrial fraction. Malondialdehyde (MDA), catalase (CAT), and glutamate peroxidase (GPx) were determined in liver homogenate. High-fat diet markedly increased the levels of AST, ALT and MDA, significantly decreasing those of SOD, CAT and GPx. But Ligusticum chuanxiong Hort-pretreatment decreased the levels of AST, ALT, and MDA. increasing those of SOD, CAT and GPx. These results demonstrated the antioxidative effects, suggesting that LCE could be the candidate for the functional material.

• Korean Journal of Veterinary Research
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• v.39 no.6
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• pp.1151-1160
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• 1999

This study was conducted to detect the toxoplasma-specific DNA in peripheral blood collected from cats experimentally infected with Toxoplasma gondii (RH strain) and from domiciled cats by B1 gene-base polymerise chain reaction(PCR). The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with parasite detection by mouse inoculation(MI). And also, latex agglutination test(LAT) and indirect fluorescent antibody test(IFAT) were conducted to detect the fluctuation of serum antibodies compared with the detection of toxoplasma by PCR and MI. Toxoplasma B1 gene PCR was shown consistently high sensitivity and the results obtained by PCR agreed completely with those from MI. All blood samples collected before infection with T gondii gave negative results by PCR and MI. Also, toxoplasma Bl gene PCR was not cross reaction with Neospora caninum DNA and normal cat leucocyte as controls. The toxoplasma-specific DNA was detected by PCR in blood of 5 cats experimentally infected with T gondii 6 days after infection and the detection of this specific-DNA was long lasted in blood for 64 days after infection. The detection of toxoplasma-specific DNA by PCR could be identified as few as 10 tachyzoites and the isolation of T gondii by MI could be isolated as few as 1 tachyzoite from tenfold serial dilution of T gondii with normal cat blood, respectively. In healthy domiciled cats, the toxoplasma-specific DNA and the parasite were detected and isolated in blood from 3 of 56(5.3%) cats by both PCR and MI, respectively. In the results of antibody test from the total 56 heads of healthy domiciled cats, the positive rates are 15(26.7%) by LAT and 19(33.9%) by IFAT. These results suggest that PCR detection of toxoplasma can be applied as a sensitive and specific diagnostic and research tool.

• Horticultural Science & Technology
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• v.33 no.3
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• pp.309-316
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• 2015

In this experiment, five commercial cultivars and one wild species of loquat were used to investigate frost tolerance and enzymatic activities in leaves and young fruits under cold stress at $-3^{\circ}C$. The frost injury, malondialdehyde (MDA) content, and oxygen-scavenging enzyme activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were studied. This results showed that the wild species 'Wild Oak-leaf' loquat was the most frost tolerant among accessions tested, followed by the cultivar 'Golden Block'. Other cultivars, 'Wu Gong Bai', 'Taicheng 4', 'Xiangzhong 11' and 'Zaozhong 6', were relatively weak in frost tolerance. The enzymatic activities of SOD, POD and CAT increased initially and then decreased as the exposure time increased. However, the enzymatic peak occurred later in the frost-tolerant accession than in the frost-sensitive accession. The correlation coefficients of MDA contents between leaves and immature fruits were from 0.93 to 0.99 in the five commercial loquat cultivars. For the 'Wild Oak-leaf' loquat, the correlation coefficients of MDA and POD were 0.98 and 0.95, respectively, but the coefficients for SOD, CAT and APX were relatively low. In general, there were good correlations between loquat leaves and immature fruits in MDA content and enzyme activities. These results indicate that analysis of these physiological and biochemical activities in loquat leaves could potentially be used to predict the cold tolerance in loquat at immature fruit stage and to accelerate breeding programs for cold tolerance in loquat.