• Journal of the Korea Academia-Industrial cooperation Society
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• v.16 no.11
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• pp.7575-7581
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• 2015

Corticotropin-releasing factor(CRF), one of the stress driven neuropeptides, was widely proposed to influence hair loss and re-growth. For the development of receptor antagonists, the screening system based on intracellular calcium signal process was developed and optimized. The aequorin parental cells were transfected with CRF1 receptor and alpha 16 promiscuous G protein cDNA to establish HEK293a16/hCRF1, a stable cell line for the human CRF1 receptor. In HEK293a16/hCRF1 cells, the range of sauvagine dose response was 12-fold higher($EC_{50}:15.21{\pm}1.83nM$) than in the transiently expressed cells, hence essential conditions for the antagonist screening experiments such as the robust signals and high solvent tolerance were secured. The standard antagonists for the CRF1 receptor, antalarmin and CP154526, resulted $IC_{50}$ values of $414.1{\pm}5.5$ and $290.7{\pm}1.9nM$, respectively. Similar results were presented with frozen HEK293a16/hCRF1 cells. Finally, our HEK293a16/hCRF1 cells with the aequorin based cellular functional assay can be a model system for the development of functional cosmetics and modulators that can have a clinical efficacy on hair re-growth.

• Journal of the Korea institute for structural maintenance and inspection
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• v.20 no.2
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• pp.122-130
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• 2016

The past few decades of industrialization enabled human-centered stream developments, which in turn resulted in constructing straight or covered streams, which are used only for sewage disposal purpose. However, these types of streams have become the cause of flood damages such as localized heavy rain. In response, various construction methods have been implemented to prevent stream and embankment damages. However, regulations regarding these measures only lay out minimum standards such as the height of slopes and the minimum angle of inclination. Moreover, examination of tractive force, the most crucial factor in preventing flood damage, is nonexistent. Therefore, this study evaluates various tractive forces by implementing a porous concrete tetrapod at a full scale artificial stream for experiment, controlling the rate of inflow, and measuring the velocity and depth of the stream under different experiment conditions. The test results of the compressive strength, and porosity and density of rock of the porous concrete tetrapod was between 16.6 and 23.2 MPa, and the actual measurement of air void was 10.1%, thus satisfying domestic standard. The result of tractive force experiment showed a limiting tractive force of $47.202N/m^2$, not satisfying the tractive force scope of $67N/m^2$ the stream design working expertise proposes. However, there was neither damage nor loss of blocks and hardpan. Based on previous researches, it can be expected that there will be resistance against a stronger tractive force. Therefore, it is necessary to conduct another experiment on practical limiting tractive force by adjusting some experimental conditions.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.200-207
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• 2018

At high salt concentration, glycine betaine is transported into Bacillus subtilis and growing rate of the cell is not suppressed. Also according to recent studies, cell growth is maintained normal growth rate at low temperature. Low temperature results in a stress response of Bacillus subtilis that is characterized by strong repression of major metabolic activities such as translation machinery and membrane transport. In this regards, genes showing cold sensitive phenotype are cold-induced DEAD box RNA helicases (ydbR, yqfR) and fatty acid desaturases (bkdR, des). Therefore to understand the effect of glycine betaine on cold growth of Bacillus subtilis, we investigated the effect of glycine betaine on growth rate of these deletion mutants showing cold sensitive phenotype. Glycine betaine strongly stimulated growth of wild type Bacillus subtilis JH642 and deletion mutants of ydbR and yqfR at $20^{\circ}C$ (190~686 min $T_d$ difference). On the other hands, glycine betaine does not show growth promoting effects on deletion mutants of bkdR, and des at cold conditions. Same cold protectant growth results were shown with the precursor choline instead of glycine betaine. We investigated the effects of detergents on the cell membrane in bkdR and des deficient strains associated with cell membrane. It was identified that bkdR deficient strain shows retarded growth with detergent such as Triton X-100 or N-lauryl sarcosine compared with wild type cell. Thus, it is possible that deletion mutation of bkdR modifies membrane structure and effects on transport of glycine betaine.

• Biomolecules & Therapeutics
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• v.31 no.1
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• pp.73-81
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• 2023

Sirtuins (SIRTs) belong to the nicotinamide adenine dinucleotide (NAD+)-dependent class III histone deacetylase family. They are key regulators of cellular and physiological processes, such as cell survival, senescence, differentiation, DNA damage and stress response, cellular metabolism, and aging. SIRTs also influence carcinogenesis, making them potential targets for anticancer therapeutic strategies. In this study, we investigated the anticancer properties and underlying molecular mechanisms of a novel SIRT1 inhibitor, MHY2251, in human colorectal cancer (CRC) cells. MHY2251 reduced the viability of various human CRC cell lines, especially those with wild-type TP53. MHY2251 inhibited SIRT1 activity and SIRT1/2 protein expression, while promoting p53 acetylation, which is a target of SIRT1 in HCT116 cells. MHY2251 treatment triggered apoptosis in HCT116 cells. It increased the percentage of late apoptotic cells and the sub-G1 fraction (as detected by flow cytometric analysis) and induced DNA fragmentation. In addition, MHY2251 upregulated the expression of FasL and Fas, altered the ratio of Bax/Bcl-2, downregulated the levels of pro-caspase-8, -9, and -3 proteins, and induced subsequent poly(ADP-ribose) polymerase cleavage. The induction of apoptosis by MHY2251 was related to the activation of the caspase cascade, which was significantly attenuated by pre-treatment with Z-VAD-FMK, a pan-caspase inhibitor. Furthermore, MHY2251 stimulated the phosphorylation of c-Jun N-terminal kinase (JNK), and MHY2251-triggered apoptosis was blocked by pre-treatment with SP600125, a JNK inhibitor. This finding indicated the specific involvement of JNK in MHY2251-induced apoptosis. MHY2251 shows considerable potential as a therapeutic agent for targeting human CRC via the inhibition of SIRT1 and activation of JNK/p53 pathway.

• Journal of Korean Tunnelling and Underground Space Association
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• v.25 no.2
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• pp.175-186
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• 2023

The buffer materials in disposal hole are exposed to the decay heat from spent nuclear fuels and groundwater inflow through adjacent rockmass. Since understanding of thermal-hydro-mechanical-chemical (T-H-M-C) interaction in buffer material is crucial for predicting their long-term performance and safety of disposal repository, it is necessary to investigate the heating-hydration characteristics and consequent T-H-M-C behavior of the buffer materials under disposal conditions considering geochemical factors. In response, the Korea Atomic Energy Research Institute developed a laboratory-scale 'Lab.THMC' experiment system, which characterizes the T-H-M behavior of buffer materials under different geochemical conditions by analyzing heating-hydration process and stress changes. This technical report introduces the detail design of the Lab.THMC system, summarizes preliminary experimental results, and outlines future research plans.

• Korean Journal of Acupuncture
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• v.41 no.2
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• pp.33-42
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• 2024

Objectives : The sigma-1 receptor is implicated in stress, depression, psychostimulant sensitization, and addiction vulnerability. Prior studies have indicated that ethanol exposure modulates sigma-1 receptor activity within the Ventral Tegmental Area (VTA). Here, we explore the sub-mechanisms underlying sigma-1 receptor activity induced by HT7 (Shinmun) stimulation in behavioral alterations following acute ethanol (ETOH) administration. Methods : Male Wistar rats were investigated for pro- and anti-inflammatory markers after injection of ETOH (1 g/kg) using cytokine enzyme-linked immunosorbent assay (ELISA)s. After confirming that HT7 stimulation changed the total distance traveled in the open field test (OFT), protein changes in the Ventral tegmental area (VTA) were measured by Western blotting. The expression level of inducible nitric oxide synthase (iNOS) after administration of a sigma-1 receptor antagonist (dihydrobromide 1047; BD1047, 10 mg/kg i.p.) and Shenmen (HT7) stimulation was compared. Results : As a result, acute ETOH administration increased proinflammatory marker levels (TNF-𝛼 and IL-6). HT7 stimulation restored the total distance response after acute ethanol administration. In addition, in the VTA, the levels of a microglial marker (iNOS), sigma-1 receptor and protein kinase C, which are predicted to be involved in up- and downregulation, were restored by HT7 stimulation. In particular, HT7 stimulation modulates iNOS expression through effects similar to BD treatment. This study suggests that the stimulatory effect of HT7 may be driven by microglial activation. Conclusions : Microglial activity is regulated by sigma-1 receptor, and sigma-1 receptor activity is regulated by HT7 stimulation. Significantly, we demonstrate that HT7 stimulation ameliorates behavioral alterations induced by acute ETOH administration through microglial activation within the VTA.

• Korean Journal of Poultry Science
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• v.37 no.1
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• pp.51-62
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• 2010

This study was conducted to investigate the effects of dietary supplementation of multiple probiotics on the performance, small intestinal microflora and immune response in laying hens and broilers. In Exp.1, a total of 800, 82 wk old Hy-line Brown$^{(R)}$ laying hens were assigned to one of the following five dietary treatment; Control, Antibiotics (avilamycin 6 ppm), Probiotics; PB-M (Micro-ferm$^{(R)}$ 0.2%), PB-L (Lacto-sacc$^{(R)}$ 0.1%), PB-Y (Y University probiotics 0.2%). Each treatment was replicated eight times with 20 birds in each replicate and two birds were housed in each cage. Twenty birds units were arranged according to completely randomized block design. Feeding trial lasted 6 wk under 16 h lighting regimen. The Exp. 2, was conducted with a total of 1,000 broilers chicks (Ross$^{(R)}$). They were divided into five treatments, same as those of Exp. 1. Birds were fed starter (0~3 wk) and grower (4~5 wk) diets. Each treatment was replicated four times with 50 birds per pen comprising of deep litter. In Exp. 1, egg production parameters, such as hen-day and hen-house egg production, egg weight, broken and soft shell egg production, feed intake and feed conversion were not significantly different among treatments. However, strength and thickness of eggshell were significantly (P<0.05) different. Among the probiotics, PB-Y showed the highest strength and thickness of eggshell. Eggshell color, egg yolk color and Haugh unit were not significantly influenced. In Exp. 2, overall weight gain (0~5 wk) and mortality were not significantly different among treatments. However, weight gain of birds from PB-Y treatment during starter (0~3 wk) was significantly lower than the birds from Control and Antibiotic treatment. During the whole period (0~5 wk), birds from Antibiotics treatment had higher feed intake and Production Index (PI) and lower feed conversion than birds from Control treatment. Probiotics treatments were not significantly different from the Control on feed intake and feed conversion. In Exp.1, there were significant (P<0.05) differences in leukocytes parameters, such as white blood cell (WBC), hetrophil (HE), lymphocytes (LY), monocyte (MO), eosinophil (EO) and stress index (SI; HE/LY) in the blood of layers. Birds from Antibiotics and probiotics treatments tended to increase these parameters. In Exp. 2, however, only SI was significantly (P<0.05) decreased in Antibiotics treatments. Concentration of serum immunoglobulin (IgG) were higher (P<0.05) in PB-M and PB-Y treatments when compared with Control treatment in Exp. 1. The population of E. coli significantly (P<0.05) decreased in birds from Antibiotics, PB-L and PB-Y treatments when compared with birds from Control treatment in Exp. 1. Metalbolizability of crude fat decreased significantly (P<0.05) in birds from probiotic treatments in Exp. 2. It was concluded that the response of probiotics on the productivity of layers and broilers were different. Probiotics increased strength and thickness of eggshell in layers, and decreased feed conversion and increased PI in broilers. Leukocytes and IgG tended to increase by supplementation of antibiotics and probiotics in layers. Intestinal E. coli tended to decrease in layers. Digestibility of crude fat of diet decreased in probiotics treatments broilers. Parameters of blood and microbial were more sensitive in layers than broilers.

• Journal of Life Science
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• v.30 no.2
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• pp.137-146
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• 2020

Calcium is one of the important secondary signaling molecules in plant cells. Calcium-dependent protein kinases (CDPK)-the sensor proteins of Ca²⁺ and phosphorylating enzymes-are the most abundant serine/threonine kinases in plant cells. They convert and transmit signals in response to various stimuli, resulting in specific responses in plants. In rice, 31 CDPK gene families have been identified, which are mainly involved in plant growth and development and are known to play roles in response to various stress conditions. However, little is known about the biochemical characteristics of CDPK proteins. In this study, OsCPK11-a CDPK in rice-was partially purified, and its biochemical characteristics were found. Partially purified OsCPK11 from rice seedlings was obtained by three-step column chromatography that involved anion exchange chromatography consisting of DEAE, hydrophobic interaction chromatography consisting of phenyl-Sepharose, and gel filtration chromatography consisting of Sephacryl-200HR. An in vitro kinase assay using partially purified OsCPK11 was also performed. This partially purified OsCPK11 had a molecular weight of 54 kDa and showed a strong hydrophobic interaction with the hydrophobic resin. In vitro kinase assay showed that the OsCPK11 also had Ca²⁺-dependent autophosphorylation activity. The OsCPK11 phosphorylated histone III-S, and the optimum pH for its kinase activity was found to be 7.5~8.0. The native OsCPK11 shared several biochemical characteristics with recombinant OsCPK11 studied previously, and both had Ca²⁺-dependent autophosphorylation activity and favored histone III-S as a substrate for kinase activity, which also had a Ca²⁺-dependence.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.169-176
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• 2008

Salicylic acid(SA) is a phytohormone that is related to plant defense mechanism. The SA accumulation is triggered by abiotic and biotic stresses. SA acts as a signal molecular compound mediating systemic acquired resistance and hypersensitive response in plant. Although the role of SA has been studied extensively, an understanding of the SA regulatory mechanism is still lacking in plants. In order to comprehend SA regulatory mechanism, we have been transformed with a SID2 promoter:GUS::LUC fusion construct into siz1-2 mutant and wild plant(Col-0). SIZ1 encodes SUMO E3 ligase and negatively regulates SA accumulation in plants. SID2(SALICYLIC ACID INDUCTION DEFICIENT2) is a crucial enzyme of SA biosynthesis. The Arabidopsis SID2 gene encodes isochorismate synthase(ICS) that controls SA level by conversion of chorismate to isochorismate. We compared the regulation of SID2 in wild-type and siz1-2 transgenic plants that express SID2 promoter:GUS::LUC constructs respectively. The expressions of $\beta$-GLUCURONIDASE and LUCIFERASE were higher in siz 1-2 transgenic plant without any stress treatment. SID2 promoter:GUS::LUC/siz1-2 transgenic plant will be used as a starting material for isolation of siz1-2 suppressor mutants and genes involved in SA-mediated stress signaling pathway.

• Korean Journal of Poultry Science
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• v.33 no.2
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• pp.141-149
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• 2006

An experiment was conducted to investigate the effects of dietary acidifier($Lactacid^(R)$) and essential oil($Immunocin^(R)$) on the performance, nutrient metabolizability, small intestinal microflora, IgG level and leukocytes and erythrocytes in broiler chickens. Five hundred males and 500 females broiler chickens($Ross^(R)$) were divided into 20 pens of 50 chickens(25 birds in each sex). Five pens were assigned to each of four dietary treatments: control, diets containing antibiotics(Bacitracin methylene disalicilate), acidifier($Lactacid^(R)$) and essential oil($Immunocin^(R)$) dietary treatments. Birds were fed experimental diets ad libitum 5 wks. Weight gain, feed intake, and feed conversion rate were significantly affected by dietary treatment(P<0.05). Overall weight gain($0{\sim}5$ wks) of $Lactacid^(R)$ treatment was significantly lower than the others. Feed intake was highest(P<0.05) in the control followed by antibiotics, $Lactacid^(R)\;and\;Immunocin^(R)$ treatment. Feed conversion rate of $Immunocin^(R)$ treatment was lowest(P<0.05) followed by antibiotics, $Lactacid^(R)$ treatment and the control. Production indices of $Immunocin^(R)$ and antibiotics treatments were significantly higher than those of the control and $Lactacid^(R)$ treatment(P<0.05). $Immunocin^(R)$ treatment was the highest and antibiotics was lowest in serum IgG level. The number of leukocytes and stress index(neutrophil/lymphocytes) tended to be lower in $Immunocin^(R)$ treatment than others. There were no significant differences in erythrocytes among the treatments. The cfu of E. coli was significantly lower in $Immunocin^(R)$ and antibiotics treatments than $Lactacid^(R)$ treatment and the control. Metabolizability of crude protein was significantly lower in the control than $Lactacid^(R)\;and\;Immunocin^(R)$ treatment while that of NFE was significantly lower in $Immunocin^(R)\;than\;Lactacid^(R)$ and antibiotics treatments. It was concluded that essential oil product $Immunocin^(R)$ is as effective as antibiotics in improving feed conversion efficiency and production index while $Lactacid^(R)$ is not.