• BMB Reports
• /
• v.31 no.4
• /
• pp.370-376
• /
• 1998

Src-Homology 2 (SH2) domains have a capacity to bind phosphotyrosine-containing sequence context and play essential roles in various cellular signaling pathways. Due to the specific nature of the binding between SH2 domains and their counterpart proteins, inhibitors of SID domain binding have drawn extensive attention as a potential candidate for therapeutic agents. Here, we describe the binding assay system to screen for the ligands or blockers of the SH2 domains with an emphasis on the $p56^{lck}$ SH2 domain. In our assay system, SID domains expressed and purified as fusion proteins to Glutathione-S-transferase (GST) were covalently attached to 96-well microtitre plates through amide bond formation, which were subsequently allowed to bind the biotinylated phosphotyrosine (pY)containing synthetic pep tides. The binding of biotinylated pY peptides was detected by the horseradish peroxidase (HRP)-conjugated streptavidin. Using the various combinations of SH2 domain-pY peptides, we observed that: (1) The binding of pY-peptides to its counterpart SH2 domain is concentration-dependent and saturable; (2) The binding is highly specific for a particular combination of SH2 domain-pY peptide pair; and (3) The binding of Lck SH2-cognate pY-peptides is specifically competed by the nonbiotinylated peptides with expected relative affinity. These results indicate that the established assay system detects the SH2-pY peptide interaction with reproducible sensitivity and specificity and is suitable for screening the specific inhibitors of $p56^{lck}$ SH2 function.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.6
• /
• pp.870-879
• /
• 2006

The present study describes the formation of succinic acid by a nonvirulent, highly osmotolerant Klebsiella pneumoniae strain SAP (succinic acid producer), its profile of metabolites, and enzymes of the succinate production pathway. The strain produced succinate along with other metabolites such as lactate, acetate, and ethanol under aerobic as well as anaerobic growth conditions. The yield of succinate was higher in the presence of $MgCO_3$ under $N_2$ atmosphere as compared with that under $CO_2$ atmosphere. Analysis of intracellular metabolites showed the presence of a smaller PEP pool than that of pyruvate. Oxaloacetate, citrate, and $\alpha$-ketoglutarate pools were considerably larger than those of isocitrate and fumarate. In order to understand the synthesis of succinate, the enzymes involved in end-product formation were studied. Levels of phosphoenolpyruvate carboxykinase, fumarate reductase, pyruvate kinase, and acetate kinase were higher under anaerobic growth conditions. Based on the profiles of the metabolites and enzymes, it was concluded that the synthesis of succinate took place via oxaloacetate, malate, and fumarate in the strain under anaerobic growth conditions. The strain SAP showed potential for the bioconversion of fumarate to succinate under $N_2$ atmosphere in the presence of $MgCO_3$. At an initial fumarate concentration of 10 g/l, 7.1 g/l fumarate was converted to 7 g/l succinate with a molar conversion efficiency of 97.3%. The conversion efficiency and succinate yield were increased in the presence of glucose. Cells grown on fumarate contained an 18-fold higher fumarate reductase activity as compared with the activity obtained when grown on glucose.

• KSBB Journal
• /
• v.5 no.3
• /
• pp.215-221
• /
• 1990

The solubilization of BSA, pepsin, trypsin and ribonuclease-a in cationic reverse micellar solutions has been investigated. For complete solubilization into reverse micellar solutions, pH values of several pH units above the isoelectric point of each protein were required. Solubilization was observed to decrease with increasing ionic strength of KCI. The size of empty micelles showed no significant change with increasing ionic strength. Trioctylmethyl ammonium chloride (TOMAC) in cyolohexane showed the lowest solubilization for the proteins. The system didodecyl dimethyl ammonium bromide (DDAB)-tetrachloroethylene was capable of solubilizing larger amounts of proteins than the system DDAB-benzene. Cetyl trimethyl ammonium bromide(CTAB)-hexanol-isooctane system showed lower solubilization than DDAB-tetrachloroethylene system, while it had higher Wo value.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.32 no.7
• /
• pp.971-975
• /
• 2003

This study was performed to find natural foods with high anti-dementia effects from natural plant resources which have been used for mental health. We screened inhibitors of prolyl endopeptidase which could be implicated in the processing of Alzheimer's disease, the principal part of dementia. One hundred natural foods were examined for prolyl endopeptidase inhibitors. Out of them, methanol extract of Acorns and Raspberry inhibited more than 90% of prolyl endopeptidase activity at 40 ppm. Also, methanol extract of Walnut inhibited 73% of prolyl endopeptidase activity at the same concentration. Therefore, the result suggests that these natural foods may have the potential to be used for preventing and treating dementia like Alzheimer disease.

• Archives of Pharmacal Research
• /
• v.18 no.1
• /
• pp.12-17
• /
• 1995

It is essential to clone the preptide transporter in order to obtain better understanding of its molecular structure, regulation, and substrate specificity. Characteristics of an endogenous peptide transporter in oocytes were studied along with expression of an exogenous protor/peptide cotransporter from rabbit intestine. And further efforts toward cloning the transporter were performed. The presence of an endogenous peptide transporter was detected in Xenopus laevis oocytes by measuring the uptake of $0.25/{mu}M(10{\;}{\mu}Ci/ml)[^3H]$-glycylsarcosine (Gly-Sar) at pH 5.5 with or without inhibitors. Yptake of Gly-Sae in oocytes was significantly inhibited by $25{\mu}M$ glycine nd sarcosine. This result suggests that a selective transporter is involved in the endogneous uptake of dipeptides. Collagenase treatment of oocytes used to strip oocytes from ovarian follicles did not affect the Gly-Sar uptake. Changing pH from 5.5 to 7.5 did not affect the Gly-Sae uptake significantly, suggesting no depedence of the endogenous transporter on a transmembrane proton gradient. An exogenous $H^+/pep-tide$ contransported was expressed after microinjection of polyadenylated messenger ribonucleic acid $[poly(A)^+ -mRNA]$ obtained from rabbit small intestine. The Gly-Sar uptake in mRNA-injected oocytes was 9 times thigher than that in water-injected oocyltes. Thus, frog occytes can be utilized fro expression cloning of the genes encoding intestinal $H^+$peptide contransporters. Size fractionation of mRNA was successfully obtained using this technique.

• Research in Plant Disease
• /
• v.25 no.1
• /
• pp.22-28
• /
• 2019

This study researched the current occurrence status of viral diseases of solanaceous crops in the main production areas of Gangwon region, for four years from 2014 to 2018. From 2014 to 2018, in eight cities and counties including Chuncheon, CMV, BBWV2, PMMoV, TSWV, and PVY occurred in chilli peppers. In case of tomato, the research was performed in 2017 and 2018 while TSWV and TYLCV were verified in Chuncheon, Hoengseong, and Cheorwon. Especially, TYLCV occurred in Chuncheon region for the first time in 2017, which was verified in 2018. Lastly, the current occurrence status of viral diseases of bell peppers was researched in seven cities and counties like Gangneung, Hoengseong, and Cheorwon from 2015 to 2018. In the results, CMV, BBWV2, PMMoV, PepMoV, and TSWV were verified in bell peppers of Gangwon region. During the research period, there were no crops suspected or infected by BWYV, so that it is judged that there is no villiferous or infected aphid by this virus in Gangwon-do. Such results of this study could be used as the basic data for the stable production of crops through the early prevention of plant diseases occurring in the solanaceous crops of Gangwon region.

• Journal of the Korean Academy of Child and Adolescent Psychiatry
• /
• v.3 no.1
• /
• pp.3-13
• /
• 1992

For the purpose of better understanding of RAD children, comparisons were performed about the developmental and psychopathological characteristics between RAD and PDD children. Study subjects were the children between the ages 2 and 6. who visitied SNUCH Child and Adolescent Psychiatry OPD or treated in Day Treatment Center during May, 1989 and Sep., 1991, diagnosed by the child psychiatrist as RAD or PDD according to DSM-III-R diagnostic criteria. A total of 40 children were included in the study. 20 were RAD(18 boys, 2 girls). 20 were PDD(20 boys). PEP was performed to all study subjects, by the raters blind to psychiatric diagnosis of each. And analysis was done about the level of developmental functioning score and psychopathological scores between RAD and PDD children. The results are as follows 'There were retardation on the general developmental functioning in RAD children, and the severity of retardation were lined up as perceptual ability>imitation>gross motor>cogitive language>eye-hand coordination>fine motor. Characteristics of psychopathology in RAD children were lined up by severity as language>play>relationship>affection >sensory. Group differences of the two shows on PDD group rejection score was significantly higher on the perception item. while RAD was superior on the perception emerging response. Differences of the psychopathology between two groups were on such areas as ; Affection, which was significantly higher on 'Absent' in RAD, while significantly higher on 'severe' in PDD children. On the interest for the play and materials. sensory and language areas, RAD children were significantly higher ratings on 'Absent' than PDD children.

• Journal of Veterinary Clinics
• /
• v.36 no.1
• /
• pp.46-52
• /
• 2019

Pimobendan has inotropic and vasodilating effects on cardiovascular system, and pentoxifylline is known to decrease blood viscosity and improve blood flow to the heart. This study investigated the pharmacokinetics and pharmacodynamics following oral administration of pimobendan-pentoxifylline powder mixture in dogs. Eight healthy dogs were included and were divided into control (n = 4) and experimental (n = 4) groups. Vehicle powder and pimobendan-pentoxifylline powder mixture (pimobendane 0.25 mg/kg, pentoxifylline 15 mg/kg) were administrated orally to control and experimental groups, respectively. Plasma samples and measurement of echocardiographic indices were obtained for 24 hours following administration. Pimobendan and pentoxifylline concentrations were investigated using liquid chromatography-mass spectrometer (LC-MS) assay. The elimination half-life ($T_{1/2}$) were $2.65{\pm}1.42hours$ for pimobendan and $0.29{\pm}0.23hours$ for pentoxifylline. The time to reach maximum concentration ($T_{max}$) were $1.08{\pm}0.72hours$ for pimobendan and $0.29{\pm}0.14hours$ for pentoxifylline. The maximum blood concentration ($C_{max}$) were $2.83{\pm}1.50ng/mL$ for pimobendan and $1184.33{\pm}932.37ng/mL$ for pentoxifylline. Among echocardiographic indices, fractional shortening (FS), left ventricular internal diameter at end systole (LVIDs), and pre-ejection period (PEP) showed significant changes at 1-4 hours after the administration of pimobendan-pentoxifylline powder mixture. No adverse effects were observed during the investigation. This study demonstrates that pimobendan-pentoxifylline powder mixture can be used to control cardiovascular diseases in dogs.

• Animal Bioscience
• /
• v.35 no.12
• /
• pp.1892-1903
• /
• 2022

Objective: A series of experiment were conducted to evaluate the effects of replacing a part of soybean meal (SBM) at 6% of broiler diets with fermented soybean meal (FSBM) obtained by single or two-stage fermentation by measuring growth performance, antioxidant activity in the jejunum and distal intestinal microflora. Methods: Soybean meal samples were prepared by single-stage fermentation using Bacillus velezensis (Bv) (FSBM_B), or Lactobacillus spp. (as commercial control) (FSBM_L). Additional SBM sample was prepared by two-stage fermentation using Bv and subsequently using Lactobacillus brevis ATCC 367 (Lb) (FSBM_B+L). Enzyme activity, chemical composition, trichloroethanoic acid-nitrogen solubility index (TCA-NSI) and antioxidant activity were measured. Then, in an in vivo study, 320 Ross308 broilers were divided into four groups with ad libitum supply of feed and water. Four groups were fed either a corn-soybean meal diet (SBM), or one of fermented SBM diets (FSBM_B+L, FSBM_B, and FSBM_L). Growth, serum characteristics, microflora, and the mRNA expression of selected genes were measured. Results: Compared to SBM, FSBM_B+L contained lower galacto-oligosaccharide, allergic protein, and trypsin inhibitor, and higher TCA-NSI by about three times (p<0.05). Reducing power and 1,1-diphenyl-2-picrylhydrazyl free radical scavenging ability correlated positively with the TCA-NSI content in FSBM. Growth performances were not significantly different among four groups. In jejunum of 35-day-old broilers, partial replacement of SBM by FSBM_B+L increased the activity of superoxide dismutase and catalase (CAT), and the FSBM_B group had the highest catalase activity (p<0.05). Partial replacement of SBM by FSBM increased relative mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and peptide transporter 1 (PepT1) (p<0.05); however, FSBM_B+L increased CAT mRNA level to 5 times of the control (p<0.05). Conclusion: Using Bv- and Lb-processed SBM through two-stage fermentation to partially replace 6% of diets will improve the gut's antioxidant activity under commercial breeding in broilers.

• Research in Plant Disease
• /
• v.23 no.3
• /
• pp.234-240
• /
• 2017

Incidence of virus diseases in red pepper of open field in Yeongyang-Gun, Gyeongbuk Province was investigated by reverse transcription-polymerase chain reaction in 2012-2016. The infection rate of Cucumber mosaic virus (CMV), Broad bean wilt virus 2 (BBWV2), Pepper mottle virus, Potato virus Y, Pepper mild mottle virus and Tomato spotted wilt virus was 46.1%, 41.5%, 2.0%, 2.0%, 4.4% and 0.1% respectively. Incidence rate of single and mixed infection was 31.2% and 62.6%. Most of single infections were CMV and BBWV2. Among mixed infections, the incidence rate of CMV+BBWV2 mixed infection was the highest as 49.3% and most of mixed infections of triplex and tetraplex included CMV+BBWV2 mixed infection. CMV single infection caused mosaic, chlorosis, yellowing and vein necrosis and BBWV2 single infection induced cholosis and mosaic. CMV+BBWV2 mixed infection caused severe mosaic with chlorosis or malformation.