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Regulatory Mechanism in Tissue-specific Expression of Insulin-like Growth Factor-I Gene (Insulin-like growth factor-I 유전자의 조직 특이적 발현에 대한 조절기전)

  • 안미라
    • KSBB Journal
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    • v.18 no.4
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    • pp.329-334
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    • 2003
  • The present study was aimed at investigating the regulatory mechanism in tissue-specific expression of insulin-like growth factor-I (IGF-I) gene. The expression of IGF-I gene was determined by a solution hybridization/RNase protection assay using total RNA prepared from rat liver or brain of various ages. The levels of IGF-I transcripts were increased in liver gradually after birth, but decreased in brain. By using an oligonucleotide (FRE) corresponding to the C/EBP binding site of the rat IGF-I exon 1, multiple forms of C/EBP${\alpha}$ and C/EBP${\beta}$ proteins, which have DNA-binding activity, were detected in the rat liver or brain. Western immunoblot and southwestern analyses show that p42$\^$C/EBP${\alpha}$/, p38$\^$C/EBP${\alpha}$/, p35$\^$C/EBP${\alpha}$/, p38$\^$C/EBP${\beta}$/, and p35$\^$C/EBP${\beta}$ form specific complexes with the IGF-I exon 1 oligonucleotide in liver nuclear extract and that p42$\^$C/EBP${\alpha}$/ and p38$\^$C/EBP${\beta}$/ form complexes in brain. These data suggest that the formation of FRE-C/EBP isoform complexes may play important roles in the tissue-specific regulation of IGF-I gene expression.

An efficient protocol for the production of transgenic Alstroemeria plants via particle bombardment

  • Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • v.47 no.1
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    • pp.66-72
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    • 2020
  • Alstroemeria plants were transformed by using an improved particle-gun-mediated transformation system. Friable embryogenic callus (FEC) induced from the leaves with axil tissues of Alstroemeria plant was used as the target tissue. Also, FEC was transformed with the bar gene was used as a selectable marker. In the case of plasmid pAHC25, 7.5% of the twice-bombarded FEC clumps showed blue foci, whereas the clumps with single bombardment showed only 2.3%. Additionally, a 90° rotation with double bombardment led to a higher frequency (6 times) of luciferase gene expression in PBL9780 than the control treatment. After 8 weeks of bombardment, more than 60 independent transgenic lines were obtained for pAHC25 and nearly 150 independent transgenic lines were obtained for PBL9780, all of which were resistant to PPT and demonstrated either GUS or luciferase activity. Regarding effect of osmotic treatment (0.2 M mannitol) with 7 different periods, the highest transient gene expression was obtained in 8 h before and 16 h after transformation in both pAHC25 and PBL9780. Compared with the control, at least three times more GUS foci and photons were observed in this treatment. With respect to different combinations of mannitol and sorbitol with 8 h before and 16 h after transformation, high numbers of transient and stable transgene expressions were observed in both 0.2 M mannitol and 0.2 M sorbitol used in the osmotic pre-culture. This combination showed the highest transformation efficiency in both pAHC25 (8.5%) and PBL9780 (14.5%). In the control treatment, only 10% of the FEC clumps produced somatic embryos. However, by using 0.2 M mannitol and 0.2 M sorbitol, the frequency of somatic embryos increased to 36.5% (pAHC25) and 22.9% (PBL9780). Of the somatic embryos produced, at least 60% germinated. Approximately 100 somatic embryos from these 210 independent transgenic lines from 2 plasmids developed into shoots, which were then transferred to the greenhouse. PCR analysis confirmed the presence of the bar gene. This is the report on the production of transgenic Alstroemeria plants by using particle bombardment with a high efficiency, thereby providing a new alternative for the transferring of gene of interests in Alstroemeria in the breeding program in the future.

Exploration of Submarine Spring Along the Coastal Areas of Busan Metropolitan City (부산 인근 연안해역에서 해저 용천수 유출 탐사)

  • Lee, Yong-Woo;Khim, Boo-Keun;Kim, Sunghan
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.18 no.4
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    • pp.178-185
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    • 2013
  • We measured salinity and $^{222}Rn$ concentration to explore submarine spring along the coastal areas (Mundongri, Icheon-ri, Jukseong-ri, Daebyeon-ri, Yeonhwa-ri, and Dadae-po) including Ilkwang Bay of Busan Metropolitan City in 2009 and 2010. Before field observation, we selected the potential and possible locations of submarine spring based on the lineament distribution and rose diagram analysis. Salinity and radon concentration were measured within the 1~2 km from the coastal lines. Radon activity decreased gradually from onshore to offshore. Vertical profiles of salinity at some stations showed lateral transport of water mass characterized by low salinity. Vertical profiles of salinity in the Ilkwang Bay, which is a unique bay in the south-eastern coastal area of Busan Metropolitan City, also showed the occurrence of low salinity in the bottom seawater. Our results suggest the possible occurrence of submarine discharge of fresh groundwater in the coastal areas around Busan Metropolitan City. In the future, intensive research should be conducted for the exploration methods of submarine spring as well for the possible utility of submarine groundwater as alternative water resources.

Phenylalanine Ammonia Lyase and Cinnamic Acid 4-Hydroxylase Activities of Rice and Pepper in response to UV and Wounding (벼와 고추에서 UV와 상처가 PAL 및 C4H 효소 활성에 미치는 영향)

  • Kim, Mi-Young;Yoon, Yong-Hwi;Lee, Jung-Hoon;Kim, Hak-Yoon;Shin, Dong-Hyun;Lee, In-Jung;Kim, Dal-Ung;Kim, Kil-Ung
    • Journal of Life Science
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    • v.12 no.3
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    • pp.274-280
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    • 2002
  • The metabolites related to phenylpropanoid pathway play an important role in the self-defense of plants and induced by environmental stress like wounding, pathogen attack, UV-irradiation and so on. The mRNA level of rite phenylalanine ammonia lyase (PAL) was increased at 12 h to 48 h, however it was gradually decreased 48 h to 60 h after UV irradiation. The PAL enzyme activities in rice were peaked at the time of 24 h after UV irradiation, on the other hand, it was not affected by wounding. The PAL enzyme activities in pepper were raised high at 24 h and 10 h by UV irradiation and wounding respectively. The cinnamic acid 4-hydroxylase (C4H) activities were increased by wounding treatment and were detected from 12 h to end time point of experiment, while UV-irradiation didn't affect the C4H activity in rice and pepper. These results were assumed that the action of isoflavonid has an alternative effect on the defenses which include wounding and UV irradiation and on the diverse roles in rice and hot pepper.

Stem cell-secreted 14,15-epoxyeicosatrienoic acid rescues cholesterol homeostasis and autophagic flux in Niemann-Pick-type C disease

  • Kang, Insung;Lee, Byung-Chul;Lee, Jin Young;Kim, Jae-Jun;Sung, Eun-Ah;Lee, Seung Eun;Shin, Nari;Choi, Soon Won;Seo, Yoojin;Kim, Hyung-Sik;Kang, Kyung-Sun
    • Experimental and Molecular Medicine
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    • v.50 no.11
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    • pp.8.1-8.14
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    • 2018
  • We previously demonstrated that the direct transplantation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) into the dentate gyrus ameliorated the neurological symptoms of Niemann-Pick type C1 (NPC1)-mutant mice. However, the clinical presentation of NPC1-mutant mice was not fully understood with a molecular mechanism. Here, we found 14,15-epoxyeicosatrienoic acid (14,15-EET), a cytochrome P450 (CYP) metabolite, from hUCB-MSCs and the cerebella of NPC1-mutant mice and investigated the functional consequence of this metabolite. Our screening of the CYP2J family indicated a dysregulation in the CYP system in a cerebellar-specific manner. Moreover, in Purkinje cells, CYP2J6 showed an elevated expression level compared to that of astrocytes, granule cells, and microglia. In this regard, we found that one CYP metabolite, 14,15-EET, acts as a key mediator in ameliorating cholesterol accumulation. In confirming this hypothesis, 14,15-EET treatment reduced the accumulation of cholesterol in human NPC1 patient-derived fibroblasts in vitro by suppressing cholesterol synthesis and ameliorating the impaired autophagic flux. We show that the reduced activity within the CYP system in the cerebellum could cause the neurological symptoms of NPC1 patients, as 14,15-EET treatment significantly rescued cholesterol accumulation and impaired autophagy. We also provide evidence that the intranasal administration of hUCB-MSCs is a highly promising alternative to traumatic surgical transplantation for NPC1 patients.

Microfluidic Assisted Synthesis of Ag-ZnO Nanocomposites for Enhanced Photocatalytic Activity (광촉매 성능 강화를 위한 미세유체공정 기반 Ag-ZnO 나노복합체 합성)

  • Ko, Jae-Rak;Jun, Ho Young;Choi, Chang-Ho
    • Clean Technology
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    • v.27 no.4
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    • pp.291-296
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    • 2021
  • Recently, there has been increasing demand for advancing photocatalytic techniques that are capable of the efficient removal of organic pollutants in water. TiO2, a representative photocatalytic material, has been commonly used as an effective photocatalyst, but it is rather expensive and an alternative is required that will fulfill the requirements of both high performing photocatalytic activities and cost-effectiveness. In this work, ZnO, which is more cost effective than TiO2, was synthesized by using a microreactor-assisted nanomaterials (MAN) process. The process enabled a continuous production of ZnO nanoparticles (NPs) with a flower-like structure with high uniformity. In order to resolve the limited light absorption of ZnO arising from its large band gap, Ag NPs were uniformly decorated on the flower-like ZnO surface by using the MAN process. The plasmonic effect of Ag NPs led to a broadening of the absorption range toward visible wavelengths. Ag NPs also helped inhibit the electron-hole recombination by drawing electrons generated from the light absorption of the flower-like ZnO NPs. As a result, the Ag-ZnO nanocomposites showed improved photocatalytic activities compared with the flower-like ZnO NPs. The photocatalytic activities were evaluated through the degradation of methylene blue (MB) solution. Scanning electron microscopy (SEM), x-ray diffraction (XRD), and energy-dispersive x-ray spectroscopy (EDS) confirmed the successful synthesis of Ag-ZnO nanocomposites with high uniformity. Ag-ZnO nanocomposites synthesized via the MAN process offer the potential for cost-effective and scalable production of next-generation photocatalytic materials.

Comparison of Yield and Workload depending on Stem Training Methods in Oriental Melon Hydroponics (참외 수경재배에서 줄기 유인 방법에 따른 수확량 및 작업 강도 비교)

  • Lee, Dong Soo;Kwon, Jin Kyung;Yun, Sung Wook;Lee, Si Young;Seo, Min Tae;Lee, Hee Ju;Lee, Sang Gyu;Kang, Tae Gyoung
    • Journal of Bio-Environment Control
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    • v.30 no.4
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    • pp.377-382
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    • 2021
  • Oriental melon (Cucumis melo L.) is generally cultivated on the ground by creeping culture. A farmer has a higher workload for training stems. This study was conducted to find out a new cultivation of oriental melon to reduce a workload and improve the quality of fruit. There were three treatments for training stem of oriental melon; upward stem growing, downward stem growing, control (creeping stem growing). The results of the plant growth and the net photosynthesis showed higher in upward stem growing. The root activity was higher in downward stem attract. The yield was not significant as 4,055kg/10a in upward stem attract and 3,983kg/10a in downward stem attract. According to the results of the ergonomic agricultural workload evaluation, in the case of the working posture, the working posture of creeping cultivation methods (squatting, bending) showed a higher risk level than the upward and downward cultivation methods. Therefore, it is recommended the upward stem attract of oriental melon is a new cultivation as well as an alternative method for creeping stem attract in terms of improving the plant growth and yield, and reducing the workload.

Expression Analysis of Chicken Interleukin-34(IL-34) for Various Pathogenic Stimulations (주요 병원균 자극에 의한 닭의 Interleukin-34 발현 분석 비교)

  • Hong, Yeong Ho
    • Korean Journal of Poultry Science
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    • v.48 no.3
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    • pp.111-122
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    • 2021
  • Recently, interleukin 34 (IL-34) was identified as the second functional ligand for macrophage colony-stimulating factor receptor (M-CSFR). IL-34 functions similarly to M-CSF through its binding to the M-CSFR. There is still insufficient information on IL-34 in chickens, which has until now been reported only through predicted sequences and not through experimental research. Thus, to confirm its expression and to determine its potent biological activity, several chicken lines and cell lines were used. Cloning of recombinant chicken IL-34 and M-CSF genes was performed to investigate their modulatory effects on proinflammatory cytokine expression in vitro. The expression levels of IL-34, M-CSF, and M-CSFR genes were upregulated in broiler chickens with leg dysfunction (cause unknown). However, IL-34 was downregulated in most pathogen-stimulated tissues. M-CSFR expression was enhanced by recombinant IL-34 and M-CSF proteins in vitro. IFN-γ expression was enhanced by recombinant IL-34, but not by M-CSF. However, IL-12 expression was not regulated in any of the treated cells, and IL-1β was decreased in all tissues. These results indicate that IL-34 and M-CSF have roles in both the classical and alternative macrophage activation pathways. Collectively, our findings demonstrate the expression of IL-34 in chickens for pathogenic trials, both in vitro and in vivo. Our results suggest that the IL-34 protein plays a role in both pro- and anti-inflammatory functions in macrophages. Therefore, further research is needed to determine the cytokines or chemokines that can be induced by IL-34 and to further elucidate the functions of IL-34 in the inflammatory pathway.

Antibacterial effect of bee venom against Gram-positive and negative bacteria isolated from mastitis in dairy cattle (봉독의 젖소 유방염 유래 그람 양성 및 음성 세균별 항균효과 분석)

  • Jung, Sukhan;Oh, Sang-Ik;Lee, Han-Gyu;Jung, Young-Hun;Hur, Tai-Young;Han, Sangmi;Baek, Kui-Jeong;Cho, Ara
    • Korean Journal of Veterinary Service
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    • v.44 no.3
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    • pp.169-174
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    • 2021
  • Mastitis is an inflammatory condition of the mammary gland, most often caused by bacterial infections, resulting in significant economic losses to the dairy industry. Antimicrobial resistance has been of great concern because of the extensive clinical use of antibiotics. For this reason, the development of new compounds as an alternative treatment to bovine mastitis is needed. Bee venom has been widely used as an oriental treatment for several inflammatory diseases and bacterial infections. The aim of the present study was to evaluate the antimicrobial activity of bee venom on bacteria isolated from bovine mastitis. A total of 107 isolates from bovine mastitic milk samples collected in 2019 and 2020 in Jeonbuk province. All bacterial isolates were tested for susceptibility to bee venom of the honey bee (Apis mellifera). In order to obtain comprehensive antibacterial activities of the bee venom, we measured the minimal inhibitory concentration (MIC) of the bee venom against bacterial strains. Bee venom showed significant inhibition of bacterial growth of Gram-negative bacteria Citrobacter spp., Escherchia coli, Klebsiella spp., Pseudomonas spp., Serratia spp. and Raoultella with MIC values of 96, 81, 72, 230, and 85 ㎍/mL, respectively, and Gram-positive bacterial Enterococcus spp., Staphylococcus spp. and Streptococcus spp. with MIC values of 29, 21 and 16 ㎍/mL, respectively. The results indicated that the MIC values were different depending on the bacterial strains, and those of Gram-positive bacteria were lower than those of Gram-negative bacteria for bee venom. These findings suggested that bee venom could be an effective antimicrobial treatment for bovine mastitis; however, further research is necessary to evaluate the mechanism underlying the antimicrobial action, its effectiveness/safety in vivo and effective application for therapeutic use.

Selection of Entomopathogenic Fungus Isaria javanica FT333 for Dual Control of Thrips and Anthracnose (총채벌레 및 고추탄저병의 동시 방제를 위한 곤충병원성 곰팡이 Isaria javanica FT333 선발)

  • Lee, Moran;Jeong, Hyeju;Kim, Jaeyoon;Kim, Dayeon;Ahn, Seung Ho;Lee, SangYeob;Han, Ji Hee
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.479-490
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    • 2018
  • Red pepper is seriously damaged by thrips (Thrips palmi) and anthracnose caused by Colletotrichum acutatum throughout its development. Because of biotic constraints, producers often depend on chemicals that are expensive and have adverse effects on the environment, operator, and beneficial insects. In addition, resistance is developed because of the repeated use of chemicals. In recent decades, the use of microorganisms in crop protection has become a credible alternative because it is eco-friendly. In this study, we aimed to select isolates with insecticidal and fungicidal activities against the pathogens that cause anthracnose and thrips. We treated T. palmi adults and juveniles with 13 strains of entomopathogenic fungi (isolated from the soil by using the insect-bait method), and 6 strains showed excellent insecticidal activity (70-100%) 5 days after the treatment. The selected isolates were cultured with C. acutatum to screen for the strain with excellent anti-fungal activities, among which an isolate FT333 showed more than 95% control efficacy against C. acutatum in vitro. The isolate was identified as Isaria javanica through its morphological characteristics and phylogenetic analysis of the ITS and ${\beta}-tubulin$ nucleotide sequences. The Isaria javanica FT333 isolate could be used effectively for dual bio-control of thrips and anthracnose during red pepper cultivation.