• Title/Summary/Keyword: life science domain

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Expression of Yippee-Like 5 (YPEL5) Gene During Activation of Human Peripheral T Lymphocytes by Immobilized Anti-CD3 (인체 말초혈액의 활성화 과정 중 yippee-like 5 (YPEL5) 유전자의 발현 양상)

  • Jun, Do-Youn;Park, Hye-Won;Kim, Young-Ho
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1641-1648
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    • 2007
  • Yippee-like proteins, which have been identified as the homolog of Drosophila yippee protein containing a zinc-finger domain, are known to be highly conserved among eukaryotes. However, their functional roles are still poorly understood. Recently we initiated ordered differential display (ODD)-polymerase chain reaction (PCR) to isolate genes of which expressions are altered following activation of human T cells. On the ODD-PCR image, one PCR-product detected in unstimulated T cells was not detectable at the time when the activated T cells traversed near $G_1/S$ boundary following activation by immobilized anti-CD3. Cloning and nucleotide sequence analysis revealed that the PCR-product was yippee-like 5 (YPEL5) gene, which was known as a human homolog of the Drosophila yippee gene. Northern blot analysis confirmed the amount of ${\sim}2.2$ kb YPEL5 mRNA expression detectable in unstimulated T cells was sustained until 1.5 hr after activation and then rapidly declined to undetectable level by 5 hr. Ectopic expression of YPEL5 gene in human cervix epitheloid carcinoma HeLa cells caused a significant reduction in cell proliferation to the level of 47% of the control. Expression of GFP-YPEL5 fusion protein in HeLa cells showed its nuclear localization. These results demonstrated that the expression level of human YPEL5 mRNA was negatively regulated in the early stage of T cell activation, and suggested that YPEL5 might exert an inhibitory effect on the cell proliferation as a nuclear protein.

The Analysis of Mental Stress using Time-Frequency Analysis of Heart Rate Variability Signal (심박변동 신호의 시-주파수 분석을 이용한 스트레스 분석에 관한 연구)

  • Seong Hong Mo;Lee Joo Sung;Kim Wuon Shik;Lee Hyun Sook;Youn Young Ro;Shin Tae Min
    • Journal of Biomedical Engineering Research
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    • v.25 no.6
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    • pp.581-587
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    • 2004
  • Conventional power spectrum methods based on FFT, AR method are not appropriate for analyzing biomedical signals whose spectral characteristics change rapidly. On the other hand, time-frequency analysis has more desirable characteristics of a time-varying spectrum. In this study, we investigated the spectral components of heart rate variability(HRV) in time-frequency domain using time frequency analysis methods. In the various time-frequency kernels functions, we studied the suitable kernels for the analysis of HRV using synthetic HRV signals. First, we evaluated the time/frequency resolution and cross term reduction of various kernel functions. Then, from the instantaneous frequency, obtained from time-frequency distribution, the method extracting frequency components of HRV was proposed. Subjects were 17 healthy young men. A coin-stacking task was used to induce mental stress. For each subjects, the experiment time was 3 minutes. Electrocardiogram, measured during the experiment, was analyzed after converted to HRV signal. In the results, emotional stress of subjects produced an increase in sympathetic activity. Sympathetic activation was responsible for the significant increase in the LF/HF ratio. Subjects were divided into two groups with task ability. Subjects who have higher mental stress have lack of task ability.

JSAP1 Interacts with Kinesin Light Chain 1 through Conserved Binding Segments (JSAP1과 Kinesin Light Chain 1의 결합 및 결합부위 규명)

  • Kim, Sang-Jin;Lee, Chul-Hee;Park, Hye-Young;Yea, Sung-Su;Jang, Won-Hee;Lee, Sang-Kyeong;Park, Yeong-Hong;Cha, Ok-Soo;Moon, Il-Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.17 no.7 s.87
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    • pp.889-895
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    • 2007
  • A conventional kinesin, KIF5/kinesin-I, is composed of two kinesin heavy chains (KHCs) and two kinesin light chains (KLCs) and binds directly to microtubules. KIF5 motor mediates the transport of various membranous organelles, but the mechanism how they recognize and bind to a specific cargo has not yet been completely elucidated. Here, we used the yeast two-hybrid system to identify the neuronal protein(s) that interacts with the tetratricopeptide repeats (TRP) of KLCI and found a specific interaction with JNK/stress-activated protein kinase-associated protein 1 (JSAP1/JIPP3). The yeast two-hybrid assay demonstrated that the TRP 1,2 domain-containing region of KLCI mediated binding to the leucine zipper domain of JSAP1. JSAP1 also bound to the TRP region of lac2 but not to neuronal KIF5A, KIF5C and ubiquitous KIF5B in the yeast two-hybrid assay. In addition, these proteins showed specific interactions in the GST pull-down assay and by co-immunoprecipitation. KLCI and KIF5B interacted with GST-ISAP1 fusion proteins, but not with GST alone. An antibody to JSAPI specifically co-immunoprecipitated KIF5s associated with JSAP1 from mouse brain extracts. These results suggest that JSAP1, as KLC1 receptor, is involved in the KIF5 mediated transport.

Cell Migration and Wound Healing Activities of Recombinant Thymosin β-4 Expressed in Escherichia coli (재조합 Thymosin β-4의 세포이동능과 상처치유능)

  • Hong, Kyo-Chang;Choi, Yung Hyun;Kim, Gun-Do;Cha, Hee-Jae;Jeon, Sung-Jong;Nam, Soo-Wan
    • Journal of Life Science
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    • v.32 no.2
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    • pp.135-141
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    • 2022
  • Thymosin β-4 (TB4) is a small peptide composed of 43 amino acids. To obtain sufficient biologically active mouse TB4 economically, we cloned and overexpressed this gene in an Escherichia coli system. With the isopropyl β-D-1-thiogalactopyranoside induction of the E. coli transformant, TB4 fusion protein with intein- and chitin-binding domain was successfully expressed in the soluble fraction within the E. coli cell. The TB4-intein - chitin-binding domain fusion protein was purified from the soluble fraction of E. coli cell lysate. The affinity chromatography with chitin beads and dithiothreitol-mediated intein self-cleavage reaction releases the TB4 peptide into the stripping solution. Sodium dodecyl sulphate - polyacrylamide gel electrophoresis and Western blot analyses were used to confirm that the recombinant TB4 peptide was produced with the expected size of 5 kDa. We found that the recombinant TB4 stimulated cell migration in the transwell plate chamber assay. After 18 hr of the treatment of the recombinant TB4 with 1 ng/ml concentration, the migration of the HT1080 cell was increased by 20% compared with that of the chemically synthesized TB4. The recombinant TB4 was also observed to promote the healing of a wound area in C57BL/6 mice by as high as 35% compared with that of the chemically synthesized TB4. These results suggest that the recombinant TB4 has better biological activity for cell migration and wound healing than that of the chemically synthesized TB4 peptide.

High-Level Production of High-Purity Human and Murine Recombinant Prion Proteins Functionally Compatible to In Vitro Seeding Assay

  • Hwang, Hae-Gwang;Kim, Dae-Hwan;Lee, Jeongmin;Mo, Youngwon;Lee, Se-Hoon;Lee, Yongjin;Hyeon, Jae Wook;Lee, Sol Moe;Cheon, Yong-Pil;Choi, Eun-Kyoung;Kim, Su Yeon;Lee, Yeong Seon;Son, Young-Jin;Ryou, Chongsuk
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1749-1759
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    • 2018
  • Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.

Effects of dietary Gelidium elegans extract on fat metabolism in preadipocyte cell and mice fed a high-fat diet (개별인정원료 우뭇가사리추출물의 체지방 감소에 관한 기능성 고찰)

  • Lee, Boo-Yong;Chung, Hee-Chul
    • Food Science and Industry
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    • v.53 no.4
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    • pp.390-396
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    • 2020
  • We investigated the anti-obesity effect of Gelidium elegans extract (GE) on 3T3-L1 preadipocytes and a high-fat-diet (HFD)-induced mouse model. The results of the present study demonstrated that GE prevents weight gain induced by a high-fat diet (HFD) by modulating the adenosine monophosphate-activated protein kinase (AMPK)-PR domain-containing 16 (PRDM16)-uncoupling protein-1 (UCP-1) pathway in a mice model. Moreover, in vitro results show that GE suppressed adipocyte differentiation by modulating adipogenic regulators, stimulated lipolysis by activating ATGL, and inhibited adipogenesis by downregulating various enzymes associated with triglyceride synthesis. GE was also found to upregulate AMPK phosphorylation as well as the expression of UCP1 and PRDM16 proteins, leading to measurable changes in the beige-like phenotype differentiation of 3T3-L1 cells. Taken together, these findings suggest the role of GE as a functional food ingredient extracted from Gelidium elegans to increase energy expenditure and anti-obesity efficacy.

Isolation and Characterization of vasa Gene of Triploid and Diploid Human Lung Flukes (Paragonimus westermani) (폐흡충의 이배체와 삼배체 vasa 유전자 분석 및 특징)

  • Lee, Keun-Hee;Yu, Hak-Sun;Hur, Jae-Won;Yu, Sung-Suk;Choi, Sun-Hee;Park, Sang-Kyun;Lee, Sun-Joo;Chung, Dong-Il;Kong, Hyun-Hee;Ock, Mee-Sun;Jeong, Hae-Jin
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.462-469
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    • 2007
  • In this study, we isolated, characterized, and compared the vasa homologous genes of diploid and triploid Paragonimus westermani and localized VASA homologous proteins in both lung fluke types. Open reading frames of Pw-vasa-2n and Pw-vasa-3n were of 1812 bp, and encoded deduced proteins of 622 amino acids with calculated molecular weights of 69.0 kDa and 68.9 kDa and pI's of 9.11 and 9.03, respectively. A comparison of these two VASA deduced protein sequences showed that only 6 of the 622 amino acids differed. The deduced sequences of Pw-VASA-2n and Pw-VASA-3n contained eight consensus sequences characteristic of the DEAD-box protein family and their N-terminal regions contained four arginine-glycine-glycine (RGG) motifs. These two lung fluke VASA-like proteins were more similar to those of other VASA proteins than to those of other DEAD-family proteins isolated from several organisms (planarian, zebra fish, mouse, and human). vasa homologous gene transcription and VASA protein expressions in triploid type lung flukes was slightly stronger than in the diploid type. Immunostaining showed that testes and a portion of the ovaries of both diploid and triploid lung flukes reacted strongly to anti-Pw-VASA antibody.

Comparison of Major and Job Satisfaction of Students and Graduates in Dental Hygiene and Nursing (대전 일부 치위생학과와 간호학과 재학생 및 졸업생의 전공과 업무 만족도 비교)

  • Baek, Seong-Min;Song, Da-Hye;Park, Jin-Seul;Bae, Jeong-Yeon;Jeong, Won-Seok;Hwang, Soo-Jeong
    • Journal of dental hygiene science
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    • v.13 no.4
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    • pp.378-385
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    • 2013
  • Although dental hygiene has haven a special domain of knowledge and its own business territory, people have misunderstandings about dental hygienists as nurses or nirses' aides in Korea. Inadequate knowledge could make negative results for university applicants to choose department of dental hygiene. This study was aimed to compare major and job satisfaction of students and graduates in dental hygiene and nursing for objective comparison and suggestion for dental hygiene. The subjects were 98 dental hygiene students, 113 nursing students, 53 dental hygienists, and 53 nurses by convenience sampling in Daejeon. The questionnaire consisted of satisfaction of college life, social identity, career path and job. ANOVA with Tukey post-hoc analysis was used with SPSS 20.0 program. Although dental hygiene students scored lower than nursing students in satisfaction of college life, social identity, career path and job, dental hygienists, nurse and dental hygiene students didn't have different satisfaction of job. Dental hygienists scored the lowest in relation with clinical work and major curriculum, and sexual discrimination. Nurses scored the lowest in working condition and salary. Dental hygienists and dental hygiene students had lower scores in information and advice of career path, entrance to graduate school, stable work, family's pride of my major, social position and so on. We suggested the community of dental hygiene should try to promote dental hygienists to public, the community of dentists should give the stable working condition, and the department of dental hygiene should improve the major curriculum to relate with clinical work.

Growth Responses of Potted Gerbera 'Sunny Lemon' under Non-Nutrient Solution Recycling System by Media and Nutrient Contents (비순환식 분화 양액재배시 배지와 양액함량에 따른 거베라 'Sunny Lemon'의 생육반응)

  • Kil, Mi-Jung;Shim, Myung-Sun;Park, Sang-Kun;Shin, Hak-Gi;Jung, Jae-A;Kwon, Young-Soon
    • Journal of agriculture & life science
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    • v.45 no.6
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    • pp.73-80
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    • 2011
  • To investigate the characteristics of plant growth and flower quality of gerbera 'Sunny Lemon' by amount of nutrient solution, young seedling plants, 'Sunny Lemon' were transplanted to rock-wool and medium of peat moss and perlite mixed with 1 to 2 and they were acclimatized in greenhouse during about 1 month. Nutrient solution supplied to the plants is sonneveld solution of 1/2 concentration and treatments launched June 24, 2010 when average plant height was $20{\pm}1cm$. Nutrient contents as a standard for starting point of irrigation by time domain reflectometry (TDR) were determined with 60-65%, 70-75%, and 80-85%. Results of growth during vegetative growth, plant height, leaf width and leaf number increased by 10% in rockwool, but they were not significantly different. As for plant growth depending on nutrient content, 80-85% treatment showed the highest values. Leaf number increased by 60%, and leaf width and plant height had a about 40% increase than initial growth. Effectiveness for flower quality, yield and days to flowering were superior when nutrient content of media was higher than in the others. Especially, average days to flowering in 80-85% content was advanced by 7-10 days compared to the day in 60-65% treatment. The total amount of nutrient supply per plant was higher in mixed medium than in rockwool, but change patterns of EC and pH were enhanced in rockwool. Based on our results, we recommended that growth, cut flower, and yield of gerbera 'Sunny Lemon' were more effective when nutrient content of mixed medium was maintained at 80-85%.

Isolation of Wild Yeasts from Soils of Reed Fields in Seocheon-gun County, Chungcheongnam-do, South Korea, and Characterization of Unrecorded Yeasts (충남 서천군 갈대밭 주변 토양에서 야생효모의 분리 및 미기록종 특성)

  • Han, Sang-Min;Lee, Sang-Yeop;Lee, Hyang-Burm;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.45 no.3
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    • pp.234-240
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    • 2017
  • The goal of this study was to isolate wild yeasts from reed fields in the Seocheon-gun county in Chungcheongnam-do, South Korea. Molecular analysis of the D1/D2 domain of the 26S rDNA of the yeasts isolated from soil samples was performed using Basic Local Alignment Search Tool (BLAST). In total, 20 yeast strains from 11 species were isolated from 20 different soil samples. Candida species were observed to be the most common yeasts. Of these 20 strains, Candida subhashii (6 strains), Candida tropicalis (3 strains), Candida boleticola MS008, and Lachancea thermotolerans (2 strains) were identified as alcohol-fermenting yeasts. Further, Bullera japonica YJ10-1, Candida subhashii J7-1, Kluyveromyces yarrowii YJ11-1, and Ustilago shanxiensis Y10-1 were newly recorded yeast strains in Korea, and therefore, their microbiological characteristics were investigated further. All of these unrecorded yeast strains had oval cells; only Candida subhashii J7-1 formed ascospores and pseudomycelia. Kluyveromyces yarrowii YJ11-1 grew in vitamin-free medium, and all of these strains, except Candida subhashii J7-1, grew in 5% NaCl-containing YPD broth.