• Title/Summary/Keyword: Protein Aggregation

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EXPRESSION OF PROTEIN KINASE C ISOFORMS IN CHEMICAL CARCINOGEN-INDUCED NEOPLASTIC TRANSFORMATION OF HUMAN EPITHELIAL CELLS (화학적 발암화에 따른 Protein Kinase C의 발현 변화)

  • Byeon, Ki-Jeong;Hong, Lak-Won;Kim, Chin-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.23 no.4
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    • pp.295-305
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    • 2001
  • Protein kinase C (PKC) is known to play a pivotal role in neoplastic transformation cells and its high expression is often found in a variety of types of tumors including oral cancer. While PKC is associated with the altered signal transduction pathway of the tumor cells, it is still unclear which isoform is involved in the carcinogenesis process. Since the cellular distributions and the roles of PKC are isoform-specific, it is very important to identify the specific target molecules to improve our understanding of the carcinogenesis processes. Thus, the present study attempted to perform chemical carcinogen-induced neoplastic transformation of human epithelial cells and analyze the specific isoform of PKCs involved in the cellular transformation. The study analyzed overall PKC responses upon MNNG(N-Methyl-N'-nitro-N-nitroso guanidine) exposure with [$^3H$] PDBu binding assay. PKC translocation was observed at high doses of MNNG treatment in the presence of extracellular calcium. Such effects were not observed in the absence of extracellular calcium. Translocational effects with exposure of MNNG was further enhanced in the presence of hydrocortisone. The result suggests that the type of PKC involved may be $Ca^{2+}$-dependent classical isoform and steroid hormone enhances PKC activation. Among cPKC isoforms examined, only $PKC-{\alpha}$ and r showed significant translocation of protein levels from cytosolic fraction to membrane fraction, as analyzed by immunoblot. $PKC-{\varepsilon}$ in nPKC class showed an inch·eased translocation, but other forms in this class did not show the effect. None of isoforms in aPKC class was affected by MNNG treatment. The study demonstrated that there was a certain specificity in the patterns of isoform induction follwong chemical carcinogen exposure and helped identify all the types of PKC isoforms expressed in human epithelial cells. It was revealed that PKC isoforms were activated in an early resonse to chemical carcinogen, suggesting that PKC be associated with carcinogenesis process from an early stage in this particular cell system. The study will contribute to improving our understanding of chemical-induced carcinogenesis in human cells and may provide a scientific basis to introduce the specific PKC inhibitors as an anticancer drug of epithelial cell-origin cancers including oral cancer.

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Expression profile of defense-related genes in response to gamma radiation stress (방사선 스트레스 반응 방어 유전자의 탐색 및 발현 분석)

  • Park, Nuri;Ha, Hye-Jeong;Subburaj, Saminathan;Choi, Seo-Hee;Jeon, Yongsam;Jin, Yong-Tae;Tu, Luhua;Kumari, Shipra;Lee, Geung-Joo
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.359-366
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    • 2016
  • Tradescantia is a perennial plant in the family of Commelinaceae. It is known to be sensitive to radiation. In this study, Tradescantia BNL 4430 was irradiated with gamma radiation at doses of 50 to 1,000 mGy in a phytotron equipped with a $^{60}Co$ radiation source at Korea Atomic Energy Research Institute, Korea. At 13 days after irradiation, we extracted RNA from irradiated floral tissues for RNA-seq. Transcriptome assembly produced a total of 77, 326 unique transcripts. In plantlets exposed to 50, 250, 500, and 1000 mGy, the numbers of up-regulated genes with more than 2-fold of expression compared that in the control were 116, 222, 246, and 308, respectively. Most of the up-regulated genes induced by 50 mGy were heat shock proteins (HSPs) such as HSP 70, indicating that protein misfolding, aggregation, and translocation might have occurred during radiation stress. Similarly, highly up-regulated transcripts of the IQ-domain 6 were induced by 250 mGy, KAR-UP oxidoreductase 1 was induced by 500 mGy, and zinc transporter 1 precursor was induced by 1000 mGy. Reverse transcriptase (RT) PCR and quantitative real time PCR (qRT-PCR) further validated the increased mRNA expression levels of selected genes, consistent with DEG analysis results. However, 2.3 to 97- fold higher expression activities were induced by different doses of radiation based on qRT-PCR results. Results on the transcriptome of Tradescantia in response to radiation might provide unique identifiers to develop in situ monitoring kit for measuring radiation exposure around radiation facilities.

Drying of Alaska Pollack in Controlled Conditions to Identify Major Factors for Textural Properties of Hwangtae (명태 건조과정에서 일어나는 수분과 수분활성도 변화가 명태건조품의 물성에 미치는 영향)

  • Ha, Ju-Yeoup;Lee, Sang-Hyun;Jung, Kyung-Jin;Jo, Young-Duk;Kim, Jae-Cherl
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1903-1907
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    • 2010
  • Drying curves for raw Alaska pollack seemed to follow typical food dehydration process with a very short initial settling down period. It was evident that there are some differences in drying rates between each part of fish body showing the highest drying rate for fish head followed by that for fish skin and that for flesh, presumably because of differences in water holding capacity of the components of each part. Specifically, the drying curve of fish flesh revealed that a boundary layer, thereby, a time period, existed which showed a big difference in moisture content and/or water activity as drying proceeds. The boundary layer in fish flesh with high moisture content between the layer contributes to reduce drying rate mainly as a consequence of protein aggregation resulting in hardening of fish flesh. The first boundary layer in this work appeared to show within several hours after initiation of drying. For Hwangtae, a naturally cyclic freeze-thaw dried and aged Alaska pollack which was popular in Korea, manufacturing process, it is clear that periodic moistening of boundary layer in fish flesh prohibits hardening fish flesh in boundary layer and enables steady moisture diffusion from inside of the fish flesh to surface of the fish body.

Valproic Acid Regulates α-Synuclein Expression through JNK Pathway in Rat Primary Astrocytes

  • Kim, Jung Nam;Kim, Min Kyeong;Cho, Kyu Suk;Choi, Chang Soon;Park, Seung Hwa;Yang, Sung-Il;Joo, So Hyun;Park, Jin Hee;Bahn, Geonho;Shin, Chan Young;Lee, He-Jin;Han, Seol-Heui;Kwon, Kyoung Ja
    • Biomolecules & Therapeutics
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    • v.21 no.3
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    • pp.222-228
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    • 2013
  • Although the role of ${\alpha}$-synuclein aggregation on Parkinson's disease is relatively well known, the physiological role and the regulatory mechanism governing the expression of ${\alpha}$-synuclein are unclear yet. We recently reported that ${\alpha}$-synuclein is expressed and secreted from cultured astrocytes. In this study, we investigated the effect of valproic acid (VPA), which has been suggested to provide neuroprotection by increasing ${\alpha}$-synuclein in neuron, on ${\alpha}$-synuclein expression in rat primary astrocytes. VPA concentration-dependently increased the protein expression level of ${\alpha}$-synuclein in cultured rat primary astrocytes with concomitant increase in mRNA expression level. Likewise, the level of secreted ${\alpha}$-synuclein was also increased by VPA. VPA increased the phosphorylation of Erk1/2 and JNK and pretreatment of a JNK inhibitor SP600125 prevented the VPA-induced increase in ${\alpha}$-synuclein. Whether the increased ${\alpha}$-synuclein in astrocytes is involved in the reported neuroprotective effects of VPA awaits further investigation.

Association between ITGB2 Genetic Polymorphisms and Tuberculosis (ITGB2 유전자 다형성과 결핵 사이의 연관성 연구)

  • Jin, Hyun-Seok;Lee, Sang-In;Park, Sangjung
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.2
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    • pp.118-125
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    • 2018
  • Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (MTB), but the genes associated with the host immune system can be attributed to the development of TB. The ITGB2 gene encodes the integrin beta 2 chain CD18 protein and is present on chromosome 21. The integrin beta 2 chain is an integrin expressed in leukocytes and plays a very important role in leukocyte maturation and attachment. ITGB2 plays an important role in the phagocytosis of MTB and the aggregation of leukocytes in MTB infections. This study examined the genetic polymorphisms of the ITGB2 gene between the TB case and normal control using Korean genomic and epidemiologic data. As a result, a statistically significant correlation was confirmed in 10 SNPs. The most significant SNP was rs113421921 (OR=0.69, CI: 0.53~0.90, $P=5.8{\times}10^{-3}$). In addition, rs173098, one of the significant 10 SNPs, is possibly located in a binding motif with the transcription factor cofactor p300, and can affect ITGB2 gene expression. These findings suggest that the pathogenesis of TB may be influenced by a range of genetic factors related to the immune function of the host, e.g., the reactions associated with the recruitment and attachment of leukocytes. The results of this study could be used to predict the infection control for tuberculosis in a patient-tailored manner.

Clinical Use of Thromboelastography as Monitor of Coagulopathy at the Pre and Post-Cardiopulmonary Bypass (개심술 환자의 체외순환 전후 혈전 탄성 묘사도의 임상적 이용)

  • 강경훈;김경훈
    • Journal of Chest Surgery
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    • v.30 no.11
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    • pp.1092-1096
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    • 1997
  • Thromboelastography(TEG) enables a global assessment of hemostatic function to be made from a single blood sample, documenting the interaction of platelets with protein coagulation cascade from the time of the initial platelet-fibrin interaction, through platelet aggregation, clot strengthening and fibrin cross linking to eventual clot Iysis. Thirty-five patients(mean age 34$\pm$ 12) undergoing open heart surgery from April 1st, 1996 to August 31th, 1996 were investigated at preoperatively and immediate, one hour, and 24 hours after cessation of cardiopulmonary bypass using TEG. Comparisons were made between classic hematological indices and TEG data. There were statistically significant correlation between maximal amplitude(MA) and platelet count before CPB, activating clotting time(ACT) and TEG date(R time, K time and a angle) at 24-hour after CPB. The data on the predictive accuracy for postoperative bleeding at 24-hour after CPB, the TEG was significantly better than ACT(57%) or the coagulation profiles(43%) as a predictor of postoperative bleeding, with an accuracy rate of 100% (P=0.0043). In conclusion, TEG seems to be easy to use, clinically accurate, cost effective and provides data which can effectively manage a patient's hemostasis.

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Improvement on Textural Properties of Soybean Curd by Freeze Denaturation of Soybeans (대두의 동결처리에 의한 두부의 텍스쳐 특성의 증진)

  • Baik, Sang-Ho;Kim, Myung-Kon;Yun, Sei-Eok;Joo, Hyun-Kyu
    • Korean Journal of Food Science and Technology
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    • v.28 no.2
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    • pp.267-272
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    • 1996
  • Effect of freezing of soybeans on instrumental and sensory textures of soybean curd was investigated. The hardness, gumminess and chewiness of soybean curd prepared with frozen soybeans were about three times as high as those prepared with unfrozen soybeans, while cohesiveness and elasticity were affected little by freezing. Sensory evaluation showed that freezing improved the quality of soybean curd. Instrumental and sensory textures of soybean curd prepared with frozen soybeans were excellent and almost same regardless of the boiling time when the soy slurry was boiled for 2.5 min or 5 min. However, the textures of soybean curd prepared with unfrozen soybeans were deteriorated by reducing the boiling time to 2.5 min. It was postulated that freezing facilitate the heat-denaturation of soyprotein to enhance aggregation of soy proteins and formation of cross-linkage between aggregate and $Ca^{++}$. Frozen soybeans resulted in soybean curd which lower fat content, while protein content of soybean curd was almost he same. Frozen soybeans gave a lower yield of soybean curd, which is supposed to be caused by the more fat loss during whey-off.

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Myxococcus xanthus socD500 mutation causes Sporulation and Induction of two C-signal Specific Genes (Myxococcus xanthus socD500에 의한 포자 형성 및 CsgA신호에 특이적 유전자의 발현에 관한 연구)

  • 이병욱
    • Journal of Life Science
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    • v.13 no.2
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    • pp.184-190
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    • 2003
  • Myxococcus xanthus is a Gram negative, rod-shaped, soil bacterium that displays a social behaviors, and multicellular development upon nutrient deprivation. The csgA gene encoding a cell surface protein is essential for developmental behaviors including rippling, aggregation, fruiting body formation and sporulation. csgA mutants show normal vegetative growth, but lack all these developmental phenotypes. Expression of the CsgA (C-signal) specific genes are eliminated or dramatically reduced in csgA mutants. In order to identify components of C-signal transduction pathway, second site mutations were introduced into csgA mutants and were identified which can fully or partially restore development of csgA mutants (Rhie, H. G. et. al. 1989. J. Bacteriol. 171, 3268-3276). One of such csgA suppressor mutations, socD500 restores only sporulation to csgA mutants at 15$^{\circ}C$. The socD500 mutaion however eliminates the three basic developmental requirements, starvation, high cell density and a solid surface. Only sporulation, not accompanied with fruiting body formation is induced simply by shifting the temperature of vegetatively growing cells from $32^{\circ}C$ to $15^{\circ}C$. Spores induced by socD500 mutation is not as thick as that of wild-type fruiting body. In socD500 genetic background, two of ten C-signal dependent genes, $\Omega$DK4506 and $\Omega$DK4406 are more highly expressed in growing cells at $15^{\circ}C$. These results indicate that the socD500 mutation may be partly involved in the regulation of expression of two C-signal dependent genes and genes for sporulation in this transduction pathway.

Gintonin influences the morphology and motility of adult brain neurons via LPA receptors

  • Kim, Do-Geun;Kim, Hyeon-Joong;Choi, Sun-Hye;Nam, Sung Min;Kim, Hyoung-Chun;Rhim, Hyewhon;Cho, Ik-Hyun;Rhee, Man Hee;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.45 no.3
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    • pp.401-407
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    • 2021
  • Background: Gintonin is an exogenous ginseng-derived G-protein-coupled lysophosphatidic acid (LPA) receptor ligand. LPA induces in vitro morphological changes and migration through neuronal LPA1 receptor. Recently, we reported that systemic administration of gintonin increases blood-brain barrier (BBB) permeability via the paracellular pathway and its binding to brain neurons. However, little is known about the influences of gintonin on in vivo neuron morphology and migration in the brain. Materials and methods: We examined the effects of gintonin on in vitro migration and morphology using primary hippocampal neural precursor cells (hNPC) and in vivo effects of gintonin on adult brain neurons using real time microscopic analysis and immunohistochemical analysis to observe the morphological and locational changes induced by gintonin treatment. Results: We found that treating hNPCs with gintonin induced morphological changes with a cell rounding following cell aggregation and return to individual neurons with time relapses. However, the in vitro effects of gintonin on hNPCs were blocked by the LPA1/3 receptor antagonist, Ki16425, and Rho kinase inhibitor, Y27632. We also examined the in vivo effects of gintonin on the morphological changes and migration of neurons in adult mouse brains using anti-NeuN and -neurofilament H antibodies. We found that acute intravenous administration of gintonin induced morphological and migrational changes in brain neurons. Gintonin induced some migrations of neurons with shortened neurofilament H in the cortex. The in vivo effects of gintonin were also blocked by Ki16425. Conclusion: The present report raises the possibility that gintonin could enter the brain and exert its influences on the migration and morphology of adult mouse brain neurons and possibly explains the therapeutic effects of neurological diseases behind the gintonin administration.

3D Bioprinted GelMA/PEGDA Hybrid Scaffold for Establishing an In Vitro Model of Melanoma

  • Duan, Jiahui;Cao, Yanyan;Shen, Zhizhong;Cheng, Yongqiang;Ma, Zhuwei;Wang, Lijing;Zhang, Yating;An, Yuchuan;Sang, Shengbo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.4
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    • pp.531-540
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    • 2022
  • Due to the high incidence of malignant melanoma, the establishment of in vitro models that recapitulate the tumor microenvironment is of great biological and clinical importance for tumor treatment and drug research. In this study, 3D printing technology was used to prepare GelMA/PEGDA composite scaffolds that mimic the microenvironment of human malignant melanoma cell (A375) growth and construct in vitro melanoma micro-models. The GelMA/PEGDA hybrid scaffold was tested by the mechanical property, cell live/dead assay, cell proliferation assay, cytoskeleton staining and drug loading assay. The growth of tumor cells in two- and three-dimensional culture systems and the anti-cancer effect of luteolin were evaluated using the live/dead staining method and the Cell Counting Kit-8 (CCK-8) method. The results showed a high aggregation of tumor cells on the 3D scaffold, which was suitable for long-term culture. Cytoskeleton staining and immunofluorescent protein staining were used to evaluate the degree of differentiation of tumor cells under 2D and 3D culture systems. The results indicated that 3D bioprinted scaffolds were more suitable for tumor cell expansion and differentiation, and the tumor cells were more aggressive. In addition, luteolin was time- and dose-dependent on tumor cells, and tumor cells in the 3D culture system were more resistant to the drug.