• Title/Summary/Keyword: 보존과학

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PDZ Domain-containing Proteins at Autotypic Junctions in Myelinating Schwann Cells (수초화 슈반세포 autotypic 세포연접의 PDZ 도메인 보유 단백질)

  • Han, Seongjohn;Park, Hyeongbin;Hong, Soomin;Lee, Donghyun;Choi, Maro;Cho, Jeongmok;Urm, Sang-Hwa;Jang, Won Hee;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.25 no.1
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    • pp.101-112
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    • 2015
  • A type of cell junction that is formed between different parts within the same cell is called autotypic cell junction. Autotypic junction proteins form tight junctions found between membrane lamellae of a cell, especially in myelinating glial cells. Some of them have postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domains, which interact with the carboxyl (C)-terminal PDZ-binding motif of other proteins. PDZ domains are protein-protein interaction modules that play a role in protein complex assembly. The PDZ domain, which is widespread in bacteria, plants, yeast, metazoans, and Drosophila, allows the assembly of large multi-protein complexes. The multi-protein complexes act in intracellular signal transduction, protein targeting, and membrane polarization. The identified PDZ domain-containing proteins located at autotypic junctions include zonula occludens-1 (ZO-1), ZO-2, pals-1-associated tight junction protein (PATJ), multi-PDZ domain proteins (MUPPs), membrane-associated guanylate kinase inverted 2 (MAGI2), and protease-activated receptor (PAR)-3. PAR-3 interacts with atypical protein kinase C and PAR-6, forming a ternary complex, which plays an important role in the regulation of cell polarity. MAGI2 interacts with ${\alpha}$-amino-3-hydroxyl-5-methyl-4-isoxazole propionate (AMPA) receptor at excitatory synapses. PATJ is detected in paranodal loops associated with claudin-1. On the other hand, MUPP1 is found in mesaxons and Schmidt-Lanterman incisures with claudin-5. ZO-1, ZO-2, and PAR-3 are found at all three sites. Different distributions of PDZ domain-containing proteins affect the development of autotypic junctions. In this review, we will describe PDZ domain-containing proteins at autotypic tight junctions in myelinating Schwann cells and their roles.

Minisatellite 5 of SLC6A18 (SLC6A18-MS5): Relationship to Hypertension and Evolutional Level (SLC6A18 유전자의 minisatellites 5 (SLC6A18-MS5)의 고혈압과의 관련성 및 진화적 의미)

  • Heo, Chang-Hwan;Lee, Sang-Yeop;Seol, So-Young;Kwon, Jeong-Ah;Jeong, Yun-Hee;Chung, Chung-Nam;SunWoo, Yang-Il
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1733-1738
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    • 2008
  • SLC6A18, one of the neurotransmitters, was reported the possible relationship to hypertension, and it contained eight blocks of minisatellites. In this study, SLC6A18-MS5 sequence which showed the highest heterozygosity among seven minisatellites was analyzed using the Transfac software, the putative binding sites for the transcription factor Pax4 and HNF4 were discovered as a result. The HNF4 is involved in the diabetes pathway and suggested the relationship to hypertension. Thus, we investigated the putative functional significance of allelic variation in this minisatellites with respect to susceptibility for hypertension. To address this possibility, we analyzed genomic DNA from the blood of 301 hypertension-free controls and 184 cases with hypertension. A statistically significant association was not identified between the allelic distribution of SLC6A18-MS5 and occurrence of hypertension. We then examined the meiotic segregation of SLC6A18-MS5 and it was transmitted following Mendelian inheritance. Therefore, this locus could be useful markers for paternity mapping and DNA fingerprinting. Moreover, we undertook a comprehensive analysis of the genomic sequence to address the evolutionary events of these variable repeats. SLC6A18 minisatellites regions are only conserved in human and primates. This result suggestedthat intronic minisatellites analysis is powerful evolution marker for the non-coding regions in primates and can provide a great insight to the molecular evolution of repeated region in primates.

Development of Broad-range and Specific 16S rRNA PCR for Use in Routine Diagnostic Clinical Microbiology (임상미생물 검출을 위한 광대한 범위와 특이도를 가지는 16S rRNA PCR법 개발)

  • Kim, Hyun-Chul;Kim, Yun-Tae;Kim, Hyogyeong;Lee, Sanghoo;Lee, Kyoung-Ryul;Kim, Young-Jin
    • Journal of Life Science
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    • v.24 no.4
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    • pp.361-369
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    • 2014
  • Broad-range and specific 16S rRNA gene PCR is used for detection and identification of bacterial pathogens in clinical specimens from patients with a high suspicion for infection. We describe the development of a broad-range and specific PCR primer, based on bacterial 16S rRNA, for use in routine diagnostic clinical microbiology services. The primers were designed by using conservative regions of 16S rRNA sequences from 10 strains. Ninety-eight clinical strains were isolated from clinical patient specimens. A total of 98 strains of bacteria were identified by phenotypic methods; PCR with newly designed primers and universal primers. All purified PCR products were sequenced using both forward and reverse primers on an automated DNA analyzer. In this study, we evaluated the usefulness of the newly designed primers and the universal primers for the detection of bacteria, and both these techniques were compared with phenotypic methods for bacteria detection. When we also tested 98 strains of clinical isolates with newly designed primers, about 778 bp DNA fragments were amplified and identified from all strains. Of the 98 strains, 94 strains (95.9%) correspond in comparison with phenotypic methods. The newly designed primers showed that the identities of 98 (100%) strains were the same as those obtained by universal PCR primers. The overall agreement between the newly designed primers and universal primers was 100%. The primer set was designed for rapid, accurate, and cheap identification of bacterial pathogens. We think the newly designed primer set is useful for the identification of pathogenic bacteria.

Microwave Vacuum Drying of Germinated Colored Rice as an Enzymic Health Food (효소식품으로서 발아유색미의 마이크로파 진공건조)

  • Kim, Suk-Shin;Kim, Sang-Yong;Noh, Bong-Soo;Chang, Kyu-Seob
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.619-624
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    • 1999
  • This work was to study the potential health food use of germinated colored rice after germinating and drying using microwave under vacuum. Colored rice was soaked in water at $15^{\circ}C$ for 2 days and then germinated at $25^{\circ}C$ for $3{\sim}4\;days$. The germinated colored rice was dried by different drying methods: microwave vacuum drying 1, microwave vacuum drying $2\;(drying{\rightarrow}crushing{\rightarrow}drying)$, hot air drying, vacuum drying and freeze drying. Each drier except freeze drier was set to maintain the sample temperature at $60^{\circ}C$. During microwave vacuum drying 1 or 2, the sample reached $60^{\circ}C$ much faster (within 5 min) and was dried much faster ($2{\sim}3\;hrs$ than the other drying methods. The initial drying rate of microwave vacuum drying was ten times faster than that of hot air drying. The microwave vacuum drying 2 retained the highest ${\alpha}-amylase$ activity, followed by microwave vacuum drying 1, freeze drying, vacuum drying, and hot air drying.

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Utilization and Recognition of Jeungpyun and Preference for Jeungpyun Containing Different Kinds of Makgeoilli (증편의 이용현황과 인지도 및 막걸리 종류를 달리한 증편의 기호도 및 구매도)

  • Lee, Gae-Soon;Kim, Gui-Sun;Park, Geum-Soon
    • Korean journal of food and cookery science
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    • v.26 no.6
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    • pp.745-752
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    • 2010
  • The purpose of this study was to investigate the perceptions and preferences held by the general public concerning jeungpyun. Self-administered questionnaires were collected from 484 residents in Daegu and Gyeongbuk. The questionnaires contained questions on the recognition of jeungpyun, purchasing, frequency, purchasing factor, and preferences for and purchasing of jeungpyun containing different kinds of makgeoilli. According to the survey, females were higher than males (31.4%) by 69.7%, and that for ages 20~30' was the highest. Most of them were students (39.7%), and 81.7% of them had an income of under 4,000,000 won per month. The results of the eating frequency rate showed that over half of the people had jeungpyun once a month, and the recognition of jeungpyun for females (3.63) was higher than that for males (3.09). The respondents who joyed eating jeungpyun mentioned 'good taste and texture' as the top reason for eating jeungpyun, and 'different food and bad flavor' was mentioned as the reason for disliking it. As a result, popularization of jeungpyun was based on two factors: 1) fundamental factor and 2) external factor, and both significantly influence the preference for and purchasing of jeungpyun,-. Therefore, future studies should be conducted to improve preference for jeungpyun as a functional food. The results of preference and purchasing show that jeungpyun prepared with rice, rubus coreanum miquel, black bean, grape and green mume makgeoilli.

Studies for Development of Freeze - Flow Process (Freeze - Flow Process 개발을 위한 기초연구)

  • Lee, Young-Chun;Shin, Dong-Bin
    • Korean Journal of Food Science and Technology
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    • v.17 no.6
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    • pp.495-499
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    • 1985
  • To develope a process by which liquid foods can be stored in the liquid state at the frozen storage temperature, suitable cryoprotectants were selected. Orange juice, chosen as an example of liquid foods, was stored with combined cryoprotectants at $-15^{\circ}C$, and quality changes of orange juice during storage were evaluated. Among 7 cryoprotectants tested, NaCl solution had lower initial freezing point than others, and initial freezing points of glucose, fructose, glycerol, propylene glycol and citric acid were close to each other. Considering flavor quality of orange juice, cryoprotectants suitable for reducing freezing point of orange juice were glucose, fructose, glycerol, and citric acid. Combined cryoprotectants for reducing freezing point of 3 and 4 folds concentrated orange juice to $-15^{\circ}C$ consisted of 10% glucose, 8% frutose, 4.6% glycerol and 3% citric acid, and 5.5% glucose, 4.5% fructose, 4.6% glycerol, and 3% cirtric acid, respectively. When destruction of ascorbic acid, sedimentation volume and sensory flavor score of orange juice stored with combined cryoprotectants at $-15^{\circ}C$ and the control stored at $-18^{\circ}C$ were compared, there were no significant differences. These results indicated that liquid foods with suitable combined cryoprotectants could be stored at $-15^{\circ}C$ or below in the liquid state without adverse effect on quality of the stored products.

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Quality Characteristics of the Mashes of Takju Prepared Using Different Yeasts (효모종류를 달리한 탁주 술덧의 품질특성)

  • Lee, Heung-Sook;Park, Chang-Sook;Choi, Jin-Young
    • Korean Journal of Food Science and Technology
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    • v.42 no.1
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    • pp.56-62
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    • 2010
  • Quality characteristics such as alcohol, acid, and sugar contents of takju brewing mashes prepared using several yeasts were investigated during 12-days of fermentation. Among the yeasts examined, S. cerevisiae led to the highest level of ethanol (10.2-13.4%) and total minor alcohols (0.729-0.831 mg/mL). Regardless of the yeasts used, the acidity showed drastic changes (pH from 3.4-4.2 down to 2.4-2.7) during the first 2 days, and displayed negligible changes from day 4. The total acid contents rapidly increased to the first 4 days of fermentation and leveled off from 4 to the end of fermentation period. The common organic acid components were lactic, succinic and acetic acids. The total and reducing sugar contents varied depending on the yeasts used, with mashes prepared using S. coreanus and S. rouxii producing the maximum total sugar contents (5.43-5.5%) at the end of fermentation. The reducing sugar showed its maximum (7.53-14.89%) at day 2, after which it decreased to its minimum levels (3.04-4.52%). The common free sugar components were glucose and fructose, while S. ellipsoideus led to a higher free sugar level (0.35-5.29%).

Genomic Organization and Promoter Characterization of the Murine Glial Cell-derived Neurotrophic Factor Inducible Transcription Factor (mGIF) Gene (생쥐 신경교세포 유래 신경영양인자 유도성 전사인자 (mGIF) 유전자의 유전체 구조 및 프로모터 특성 분석)

  • Kim, Ok-Soo;Kim, Yong-Man;Kim, Nam-Young;Lee, Eo-Jin;Jang, Min-Kyung;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.167-173
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    • 2007
  • To study the transcriptional mechanisms by which expression of the murine glial cell-derived neurotrophic factor inducible transcription factor (mGIF) gene is regulated, a murine genomic clone was iso-lated using a mGIF cDNA as probe. A 13-kb genomic fragment, which comprises 4-kb upstream of the transcription initiation site was sequenced. The promoter region lacks a TATA box and CAAT box, is rich in G+C content, and has multiple putative binding sites for the transcription factor Spl. The mGIF gene also has consensus sequences for AP2 binding sites. The transcriptional activity of five deletion mutants of a 2.1-kb fragment was analyzed by modulating transcription of the heterologous luciferase gene in the promoterless plasmid pGL2-Basic. All mutants showed significant transcriptional activity in the murine neuroblastoma cell line NB41A3. Transient expression assays suggested the presence of a positive regulator between -213 and -129 while a negative regulator was found in the region between -806 and -214. Relatively strong transcriptional activity was observed in neuronal NB41A3, glial C6 cells and hepatic HepG2, but very weak activity in skeletal muscle C2C12 cells. These findings confirm the tissue-specific activity of the mGIF promoter and suggest that this gene shares structural and functional similarities with the dopamine receptor genes that it regulates.

MicroRNA-23b is a Potential Tumor Suppressor in Diffuse Large B-cell Lymphoma (미만성 거대 B 세포 림프종(DLBCL)에서 microRNA-23b의 잠재적 종양 억제자로서의 효과)

  • Nam, Jehyun;Kim, Eunkyung;Kim, Jinyoung;Jeong, Dawoom;Kim, Donguk;Kwak, Bomi;Kim, Sang-Woo
    • Journal of Life Science
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    • v.27 no.2
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    • pp.149-154
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    • 2017
  • Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-hodgkin lymphoma. Advances in the chemotherapeutic treatment of this disease have improved the outcomes of DLBCL; nonetheless, many patients still die of DLBCL, and therefore, a better understanding of this disease and identification of novel therapeutic targets are urgently required. In a recent gene expression profiling study, PDE (phosphodiesterase) 4B was found to be overexpressed in chemotherapy-resistant tumors. The major function of PDE4B is to inactivate the second messenger cyclic 3',5' monophosphate (cAMP) by catalyzing the hydrolysis of cAMP to 5'AMP. It is known that cAMP induces cell cycle arrest and/or apoptosis in B cells, and PDE4B abolishes cAMP's effect on B cells. However, the mechanism by which PDE4B is overexpressed remains unclear. Here, we show that the aberrant expression of miRNA may be associated with the overexpression of this gene. The PDE4B 3' untranslated region (UTR) has three functional binding sites of miR-23b, as confirmed by luciferase reporter assays. Interestingly, miR-23b-binding sites were evolutionarily conserved from humans to lizards, implying the critical role of PDE4B-miR-23b interaction in cellular physiology. The ectopic expression of miR-2 3b repressed PDE4B mRNA levels and enhanced intracellular cAMP concentrations. Additionally, miR-23b expression inhibited cell proliferation and survival of DLBCL cells only in the presence of forskolin, an activator of adenylyl cyclase, suggesting that miR-23b's effect is via the downregulation of PDE4B. These results together suggest that miR-23b could be a therapeutic target for overcoming drug resistance by repressing PDE4B in DLBCL.

Development and Validation of Real-time PCR to Determine Branchiostegus japonicus and B. albus Species Based on Mitochondrial DNA (Real-time PCR 분석법을 이용한 옥돔과 옥두어의 종 판별법 개발)

  • Chung, In Young;Seo, Yong Bae;Yang, Ji-Young;Kim, Gun-Do
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1331-1339
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    • 2017
  • DNA barcoding is the identification of a species based on the DNA sequence of a fragment of the cytochrome C oxidase subunit I (COI) gene in the mitochondrial genome. It is widely applied to assist with the sustainable development of fishery-product resources and the protection of fish biodiversity. This study attempted to verify horse-head fish (Branchiostegus japonicus) and fake horse-head fish (Branchiostegus albus) species, which are commonly consumed in Korea. For the validation of the two species, a real-time PCR method was developed based on the species' mitochondrial DNA genome. Inter-species variations in mitochondrial DNA were observed in a bioinformatics analysis of the mitochondrial genomic DNA sequences of the two species. Some highly conserved regions and a few other regions were identified in the mitochondrial COI of the species. In order to test whether variations in the sequences were definitive, primers that targeted the varied regions of COI were designed and applied to amplify the DNA using the real-time PCR system. Threshold-cycle (Ct) range results confirmed that the Ct ranges of the real-time PCR were identical to the expected species of origin. Efficiency, specificity and cross-reactivity assays showed statistically significant differences between the average Ct of B. japonicus DNA ($21.85{\pm}3.599$) and the average Ct of B. albus DNA ($33.49{\pm}1.183$) for confirming B. japonicus. The assays also showed statistically significant differences between the average Ct of B. albus DNA ($22.49{\pm}0.908$) and the average Ct of B. japonicus DNA ($33.93{\pm}0.479$) for confirming B. albus. The methodology was validated by using ten commercial samples. The genomic DNA-based molecular technique that used the real-time PCR was a reliable method for the taxonomic classification of animal tissues.