• Title/Summary/Keyword: C-Jun expression

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Association of SNPs from iNOS and TLR-4 Genes with Economic Trait in Chicken (닭의 iNOS와 TLR-4 유전자 내 변이와 경제 형질 간의 연관성 분석)

  • Lim, Hee Kyong;Han, Jung-Min;Oh, Jae Don;Lee, Hak Kyo;Jeon, Gwang Joo;Lee, Jun Heon;Seo, Dong Won;Cahyadi, Muhammad;Song, Ki Duk;Choi, Kang Duk;Kong, Hong Sik
    • Korean Journal of Poultry Science
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    • v.40 no.2
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    • pp.83-89
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    • 2013
  • iNOS (Inducible nitric oxide synthase) and TLR-4 (Toll-like Receptor-4) play crucial roles in innate immunity of poultry. iNOS has been mapped to chicken chromosome 14 and implicated in a variety of chicken diseases. iNOS possesses potent antimicrobial activity, including the inhibition of microbes replication in vitro. TLR-4 is a pathogen associated molecular-pattern receptor for bacterial product, such as LPS (lipopolysaccharides) found in Gram negative bacteria, that triggers pro-inflammatory cytokine expression after engagement with ligands. In the previous studies, genetic analysis of iNOS and TLR-4 revealed the possible association of mutation in these genes with the intestinal microflora of cecum when infected with Salmonella spp. This study was aimed to augment previous findings, which show the association of iNOS (C14513T) and TLR-4 (G4409T) polymorphisms with economic traits in Korean Native Black (KNB), Rhode Island Red (RIR) and Cornish chickens. Investigation in the effect of SNPs on economic traits (layday, layw, layno, bw150, bw270, layw270) was conducted. iNOS (C14513T) had a significant effect on the average body weight at 270 days of age (p<0.05) in both KNB and RIR, whereas TLR-4 (G4409T) showed no significant correlation with all traits (p>0.05). The results obtained from using the candidate genes can be useful for the genetic improvement of body weight in both KNB and RIR breeds.

Leaf Shape Index, Growth, and Phytochemicals in Two Leaf Lettuce Cultivars Grown under Monochromatic Light-emitting Diodes (단색 발광다이오드에서 자란 축면상추 두 품종의 엽형, 생장 및 기능성 물질)

  • Son, Ki-Ho;Park, Jun-Hyung;Kim, Daeil;Oh, Myung-Min
    • Horticultural Science & Technology
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    • v.30 no.6
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    • pp.664-672
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    • 2012
  • As an artificial light source, light-emitting diode (LED) with a short wavelength range can be used in closed-type plant production systems. Among various wavelength ranges in visible light, individual light spectra induce distinguishing influences on plant growth and development. In this study, we determined the effects of monochromatic LEDs on leaf shape index, growth and the accumulation of phytochemicals in a red leaf lettuce (Lactuca sativa L. 'Sunmang') and a green leaf lettuce (Lactuca sativa L. 'Grand rapid TBR'). Lettuce seedlings grown under normal growing conditions ($20^{\circ}C$, fluorescent lamp + high pressure sodium lamp, $130{\pm}5{\mu}mmol{\cdot}m^{-2}{\cdot}s^{-1}$, 12 hours photoperiod) for 18 days were transferred into incubators at $20^{\circ}C$ equipped with various monochromatic LEDs (blue LED, 456 nm; green LED, 518 nm; red LED, 654 nm; white LED, 456 nm + 558 nm) under the same light intensity and photoperiod ($130{\pm}7{\mu}mmol{\cdot}m^{-2}{\cdot}s^{-1}$, 12 hours photoperiod). Leaf length, leaf width, leaf area, fresh and dry weights of shoots and roots, shoot/root ratio, SPAD value, total phenolic concentration, antioxidant capacity, and the expression of a key gene involved in the biosynthesis of phenolic compounds, phenylalanine ammonia-lyase (PAL), were measured at 9 and 23 days after transplanting. The leaf shape indexes of both lettuce cultivars subjected to blue or white LEDs were similar with those of control during whole growth stage. However, red and green LEDs induced significantly higher leaf shape index than the other treatments. The green LED had a negative impact on the lettuce growth. Most of growth characteristics such as fresh and dry weights of shoots and leaf area were the highest in both cultivars subjected to red LED treatment. In case of red leaf lettuce plants, shoot fresh weight under red LED was 3.8 times higher than that under green LED at 23 days after transplanting. In contrast, the accumulation of chlorophyll, phenolics including antioxidants in lettuce plants showed an opposite trend compared with growth. SPAD value, total phenolic concentration, and antioxidant capacity of lettuce grown under blue LED were significantly higher than those under other LED treatments. In addition, PAL gene was remarkably activated by blue LED at 9 days after transplanting. Thus, this study suggested that the light quality using LEDs is a crucial factor for morphology, growth, and phytochemicals of two lettuce cultivars.

Lactate Dehydrogenase and Monocarboxylate Transporters 1, 2, and 4 in Tissues of Micropterus salmoides (큰입우럭(Micropterus salmoides) 조직의 젖산탈수소효소 및 Monocarboxylate 수송체(MCT) 1, 2, 4)

  • Yum, Jung-Joo;Yeon, Jun-Hee
    • Journal of Life Science
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    • v.22 no.1
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    • pp.98-109
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    • 2012
  • The properties of lactate dehydrogenase (EC 1.1.1.27, LDH) and expression of monocarboxylate transporters (MCTs) 1, 2, and 4 were studied in tissues from Micropterus salmoides. Native-PAGE revealed that the LDH $A_4$ isozyme was predominantly located in skeletal muscle. The LDH $A_4$, $A_2B_2$, and $B_4$ isozymes were detected in heart, liver, eye, and brain tissues, while eye-specific $C_4$ isozyme was detected in eye tissue. In September, strong LDH $B_4$ isozyme activity was detected in heart tissue. High $A_4$ isozyme activity was noted in all other tissues except heart tissue. However, in November, strong $A_4$ isozyme activity was detected in heart tissue. The LDH/CS (Citrate synthase, EC 4.1.3.7) ratio in skeletal muscle and heart tissues indicated that anaerobic metabolism was high in those tissues. Native-PAGE after immunoprecipitation showed that eye-specific $C_4$ isozyme was more similar to the $A_4$ than the $B_4$ isozyme. The LDH $A_4$ isozyme was purified by affinity chromatography. The molecular weight of subunit A was 37,200. The LDH activity in tissues was consistently 11.05~28.32% due to inhibition by 10 mM pyruvate. The $K_m^{PYR}$ of LDH in eye tissue was very low. The optimum pH for LDH in tissues was pH 7.5~8.0. The LDH $A_4$ isozyme was detected in mitochondria of skeletal muscle, whereas the $B_4$ and $A_2B_2$ isozymes were detected in heart tissue mitochondria. Western blot analysis indicated that MCTs 1, 2, and 4 were located in the plasma membrane and mitochondria of skeletal muscle and heart tissues. The sizes of MCTs 1, 2, and 4 in skeletal muscle were 60, 54~38, and 63 kDa, while those in heart tissue were 57, 54~38, and 55.5 kDa, respectively. In conclusion, M. salmoides appears to use anaerobic metabolism predominantly when adapted to a hypoxic environment. In highly activated skeletal muscle and heart tissue, energy production is controlled by inward and outward flows of pyruvate and lactate through MCTs 1, 2, and 4 in the plasma membrane and mitochondria, with effective adjustment by LDH isozymes.

Effect of Multidrug Resistance Gene-1 (mdr1) Overexpression on In-Vitro Uptake of $^{99m}Tc$-sestaMIBl in Murine L1210 Leukemia Cells (백혈병 세포에서 Multidrug Resistance Gene-1 (mdr1)의 과발현이 $^{99m}Tc$-sestaMIBl 섭취에 미치는 영향)

  • Chun, Kyung-Ah;Lee, Jae-Tae;Lee, Sang-Woo;Kang, Do-Young;Sohn, Sang-Kyun;Lee, Jong-Kee;Chung, June-Key;Jun, Soo-Han;Lee, Kyu-Bo
    • The Korean Journal of Nuclear Medicine
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    • v.33 no.2
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    • pp.152-162
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    • 1999
  • Purpose: To determine whether $^{99m}Tc$-MIBI is recognized by the multidrug resistant P-glycoprotein (Pgp), we have measured quantitatively $^{99m}Tc$-MIBI uptake in cancer cells. The effects of various Pgp reversing agents on cellular $^{99m}Tc$-MIBI uptake were also investigated in the presence of multidrug resistance gene-1 (mdr1 gene) overexpression. Materials and Methods: We measured percentage uptake of $^{99m}Tc$-MIBI at different incubation temperatures both in mdr1 positive and negative cells. The effects of verapamil, cyclosporin, and dipyridamole on cellular uptake of $^{99m}Tc$-MIBI were also evaluated with or without overex-pression of mdr1 gene in cultured murine leukemia Ll210 cells. Results: The mdr1 gene expressing cell lines were effectively induced in in vitro with continuous application of low-dose adriamycin or vincristine. Cellular uptake of $^{99m}Tc$-MIBI was higher in mdr1 negative Ll210 cells than those of mdr1 positive cells, and higher when incubated in $37^{\circ}C$ than $4^{\circ}C$. In the presence of verapamil, cyclosporin or dipyridamole, $^{99m}Tc$-MIBI uptake was increased upto 604% in mdr1 positive cells. Conclusion: Cellular uptake of $^{99m}Tc$-MIBI is lower in leukemia cells over-expressing mdr1 gene, and MBR-reversing agents increase cellular uptake. These results suggest that $^{99m}Tc$-MIBI can be used for characterizing Pgp expression and developing MDR-reversing agents in vitro.

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Inhibitory Effects of Filiform Acupuncture and Laser Acupuncture at the Points of Zhongzhu($TE_3$).Zulinqi($GB_{41}$) on Neuropathic Pain in the Tibial and Sural Nerve Injury-induced Rats (중저(中渚)($TE_3$).임입(臨泣)($GB_{41}$) 침자 및 레이저침이 백서의 Tibial Nerve와 Sural Nerve 유발 동통 억제에 미치는 영향)

  • Lee, Ju Hee;Lee, Dong Geun;Lee, Ook Jae;Lee, Sang Hyun;Lee, Jung Hun;Jeong, Joo Yong;Cheong, Min Seong;Yang, Tae Jun;Kim, Seon Wook;Cho, Myoung Rae
    • Journal of Acupuncture Research
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    • v.31 no.2
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    • pp.119-134
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    • 2014
  • Objectives : The purpose of this study was to investigate the analgesic effects of filiform acupuncture and laser acupuncture at the points of $TE_3{\cdot}GB_{41}$ in neuropathic pain induced rats. Methods : Neuropathic pain in rats was induced by partial resection of the tibial and sural nerves. Three weeks after the neuropathic surgery, each of the experimental groups(AT, LAT, AT + LAT and LAT + AT) was injected at the $TE_3{\cdot}GB_{41}$ twice per week for three weeks. Results : 1. All of the experimental groups(AT, LAT, AT + LAT and LAT + AT) showed a significant decrease in the plantar withdrawal response of allodynia and the thermal allodynia as compared with the control group. During the early phase, the AT and AT + LAT groups have been marked as more significant than the LAT + AT and LAT groups. 2. The expression of c-Fos significantly decreased in the LAT and LAT + AT groups as compared with the control group. 3. The LAT + AT group showed a significant decrease in Bax as compared with the control group. In each experimental groups(AT, LAT, AT + LAT and LAT + AT), Bcl-2 increased and Bax/Bcl-2 ratio decreased as compared with the control group. 4. The LAT, AT + LAT and LAT + AT groups showed a significant increase in mGluR5 as compared with the control group. Conclusions : These results represented that the filiform acupuncture and laser acupuncture at the $TE_3{\cdot}GB_{41}$ exerted anti-apoptotic and neuroprotective effects on the model of neuropathic pain, thereby suggesting that they should be available for decreasing mechanical allodynia.

Substrate roughness induces the development of defective E-cadherin junctions in human gingival keratinocytes

  • Jin, Chengbiao;Lee, Gayoung;Oh, Changseok;Kim, Hyun Jung;Kim, Hyun-Man
    • Journal of Periodontal and Implant Science
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    • v.47 no.2
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    • pp.116-131
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    • 2017
  • Purpose: The entry of bacteria or harmful substances through the epithelial seal of human gingival keratinocytes (HGKs) in the junctional epithelium (JE) is blocked by specialized intercellular junctions such as E-cadherin junctions (ECJs). However, the influence of roughened substrates, which may occur due to apical migration of the JE, root planing, or peri-implantitis, on the development of the ECJs of HGKs remains largely unknown. Methods: HGKs were cultured on substrates with varying levels of roughness, which were prepared by rubbing hydrophobic polystyrene dishes with silicon carbide papers. The activity of c-Jun N-terminal kinase (JNK) was inhibited with SP600125 or by transfection with JNK short hairpin RNA. The development of intercellular junctions was analyzed using scanning electron microscopy or confocal laser scanning microscopy after immunohistochemical staining of the cells for E-cadherin. The expression level of phospho-JNK was assessed by immunoblotting. Results: HGKs developed tight intercellular junctions devoid of wide intercellular gaps on smooth substrates and on rough substrates with low-nanometer dimensions (average roughness $[Ra]=121.3{\pm}13.4nm$), although the ECJs of HGKs on rough substrates with low-nanometer dimensions developed later than those of HGKs on smooth substrates. In contrast, HGKs developed short intercellular junctions with wide intercellular gaps on rough substrates with mid- or high-nanometer dimensions ($Ra=505.3{\pm}115.3nm$, $867.0{\pm}168.6nm$). Notably, the stability of the ECJs was low on the rough substrates, as demonstrated by the rapid destruction of the cell junction following calcium depletion. Inhibition of JNK activity promoted ECJ development in HGKs. JNK was closely associated with cortical actin in the regulation of ECJs in HGKs. Conclusions: These results indicate that on rough substrates with nanometer dimensions, the ECJs of HGKs develop slowly or defectively, and that this effect can be reversed by inhibiting JNK.

Diallyl Disulfide Prevents Cyclophosphamide-Induced Hemorrhagic Cystitis in Rats through the Inhibition of Oxidative Damage, MAPKs, and NF-κB Pathways

  • Kim, Sung Hwan;Lee, In Chul;Ko, Je Won;Moon, Changjong;Kim, Sung Ho;Shin, In Sik;Seo, Young Won;Kim, Hyoung Chin;Kim, Jong Choon
    • Biomolecules & Therapeutics
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    • v.23 no.2
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    • pp.180-188
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    • 2015
  • This study investigated the possible effects and molecular mechanisms of diallyl disulfide (DADS) against cyclophosphamide (CP)-induced hemorrhagic cystitis (HC) in rats. Inflammation response was assessed by histopathology and serum cytokines levels. We determined the protein expressions of nuclear transcription factor kappa-B (NF-${\kappa}B$), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), oxidative stress, urinary nitrite-nitrate, malondialdehyde (MDA), and 8-hydroxy-2'-deoxyguanosine (8-OHdG). Finally, we studied the involvement of mitogen-activated protein kinases (MAPKs) signaling in the protective effects of DADS against CP-induced HC. CP treatment caused a HC which was evidenced by an increase in histopathological changes, proinflammatory cytokines levels, urinary nitrite-nitrate level, and the protein expression of NF-${\kappa}B$, COX-2, iNOS, TNF-${\alpha}$, p-c-Jun N-terminal kinase (JNK), and p-extracellular signal regulated kinase (ERK). The significant decreases in glutathione content and glutathione-S-transferase and glutathione reductase activities, and the significant increase in MDA content and urinary MDA and 8-OHdG levels indicated that CP-induced bladder injury was mediated through oxidative DNA damage. In contrast, DADS pretreatment attenuated CP-induced HC, including histopathological lesion, serum cytokines levels, oxidative damage, and urinary oxidative DNA damage. DADS also caused significantly decreased the protein expressions of NF-${\kappa}B$, COX-2, iNOS, TNF-${\alpha}$, p-JNK, and p-ERK. These results indicate that DADS prevents CP-induced HC and that the protective effects of DADS may be due to its ability to regulate proinflammatory cytokines production by inhibition of NF-${\kappa}B$ and MAPKs expressions, and its potent anti-oxidative capability through reduction of oxidative DNA damage in the bladder.

Association Study between 2 Single Nucleotide Polymorphisms in Estrogen Receptor 1 Gene and Korean Female Schizophrenic Patients (Estrogen Receptor 1 유전자 내 2개의 단일염기 다형성과 한국인 여성 정신분열병 환자의 연합에 관한 연구)

  • Park, Jin Kyung;Lee, Seo Kyong;Lee, Sang Min;Park, Jun Heon;Kim, Ji Yeong;Cho, Ah Rang
    • Korean Journal of Biological Psychiatry
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    • v.14 no.4
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    • pp.241-248
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    • 2007
  • Objectives : Schizophrenia is equally distributed in both sexes. However, later-onset, milder psychopathology, and better outcome are associated with the females. This reason is thought to be partly due to the estrogen system. Recently, it was suggested that estrogen receptor 1(ESR1) gene polymorphisms might affect the expression of ESR1 and were associated with several psychiatric disorders. Thus, we investigated the association between two single nucleotide polymorphisms(SNPs) in the ESR1 gene and Korean schizophrenic patients in this study. Methods : Genotype, allele, and haplotype frequencies of the two SNPs(rs 2234693 and rs 2228480) were analyzed between 218 Korean controls and 158 Korean schizophrenic patients. Also, age of onset and negative symptom scale scores according to genotypes were analyzed in the patients with schizophrenia. Results : There was a significant difference in allele frequencies of rs 2234693 between the schizophrenic patients and the controls(p=0.03). Genotype distributions(p=0.03) and allele frequencies(p=0.01) of rs 2234693 were significantly different between the female schizophrenic patients and the female controls. The frequency of TC-CC genotypes compared with TT genotype in the female schizophrenic patients was significantly higher than that in the female controls(OR=2.36). The mean age of onset in the schizophrenic patients with TC-CC genotypes was significantly lower than that in the patients with TT genotype. The frequency of rs 2234693C- rs 2228480G haplotype in the female schizophrenic patients was relatively higher than that in the female controls. Conclusions : These results of our study support the possibility that the ESR1 gene polymorphisms might be involved in the susceptibility of females to schizophrenia and play a role in sex difference of schizophrenia.

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Molecular Cloning and Functional Expression of Extracellular Exo-β-(1,3)-Glucanase from Candida fermentati SI (Candida fermentati SI의 exo-β-(1,3)-glucanase유전자의 클로닝 및 그 특성)

  • Lim, Yu-Mi;Kim, Bong-Ki;Kim, Sang-Jun;So, Jai-Hyun;Kim, Won-Chan
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.317-323
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    • 2016
  • An isoflavone glucosidase that catalyzes the hydrolysis of isoflavone glucosides into glucose and corresponding aglycones was purified from Candida fermentati SI. The N-terminal sequence was determined to be GLNCDYCN. We designed degenerate primers on the basis of these amino acid sequences and successfully cloned the full structural gene sequence of the isoflavone glucosidase using inverse PCR. The exo-β-(1,3)-glucanase gene consists of 1227 base-pair nucleotides, encoding a 408-amino-acid sequence that shares 41–96% amino acid homology with other yeast exo-β-(1,3)-glucanases belonging to glycoside hydrolase family 5. The recombinant exo-β-(1,3)-glucanase was expressed in Pichia pastoris X-33, using a pPICZA vector system, and further characterized. The molecular mass of the purified exo-β-(1,3)-glucanase was estimated by SDS-PAGE to be 47 kDa. The optimal pH and temperature were pH 4.5 and 40℃, respectively. The Km values of the purified exo-β-(1,3)-glucanase for daidzin and genistin were 0.12 mM and 0.14 mM, respectively. The Vmax values of the purified isoflavone glucosidase were 945.03 U/mg for daidzin and 835.92 U/mg and for genistin.

Polysaccharide isolated from fermented barley extract activates macrophages via the MAPK and NF-κB pathways (보리발효추출물로부터 분리한 다당의 대식세포 활성화 및 신호 전달)

  • Kim, Han Wool;Jee, Hee Sook;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.50 no.5
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    • pp.555-563
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    • 2018
  • Barley has nutritional benefits due to its high dietary fiber content; therefore, the intake of whole barley grains is recommended. However, barley is often consumed in the fermented form because of the improved texture and digestibility. The present study was designed to elucidate the intracellular signaling pathway for macrophage activation by the polysaccharide BF-CP from fermented barley. BF-CP is a neutral polysaccharide, composed of neutral sugars, including glucose (70.7%), xylose (11.4%), and arabinose (9.0%). BF-CP exhibited macrophage-stimulatory activity by inducing the production of interleukin (IL)-6, tumor necrosis factor $(TNF)-{\alpha}$, and nitric oxide in RAW 264.7 macrophages. Further, BF-CP treatment strongly increased the IL-6 and $TNF-{\alpha}$ gene expression in a concentration-dependent manner. Signal transduction experiments using immunoblotting showed that BF-CP phosphorylated mitogen-activated protein kinases (MAPKs), such as c-Jun N-terminal kinase, extracellular signal-regulated kinase, and p38, and nuclear factor $(NF)-{\kappa}B$, in RAW 264.7 cells in a concentration-dependent manner. These results suggest that BF-CP activates the macrophages via MAPK and $NF-{\kappa}B$ pathways, and also induces an increase in the production of cytokines.