• Title/Summary/Keyword: molecular data

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Korea Brassica Genome Project: Current Status and Prospective (배추 유전체열구의 현황과 전망)

  • Choi, Su-Ryun;Park, Jee-Yong;Park, Beom-Seok;Kim, Ho-Il;Lim, Yong-Pyo
    • Journal of Plant Biotechnology
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    • v.33 no.3
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    • pp.153-160
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    • 2006
  • Brassica rape is an important species used as a vegetable, oil, and fodder worldwide. It is related phylogenically to Arabidopsis thaliana, which has already been fully sequenced as a model plant. The 'Multinational Brassica Genome Project (MBGP)'was launched by the international Brassica community with the aim of sequencing the whole genome of B. rapa in 2003 on account of its value and the fact that it has the smallest genome among the diploid Brassica. The genome study was carried out not only to know the structure of genome but also to understand the function and the evolution of the genes comprehensively. There are two mapping populations, over 1,000 molecular markers and a genetic map, 2 BAC libraries, physical map, a 22 cDHA libraries as suitable genomic materials for examining the genome of B. rapa ssp. pekinensis Chinese cabbage. As the first step for whole genome analysis, 220,000 BAC-end sequences of the KBrH and KBrB BAC library are achieved by cooperation of six countries. The results of BAC-end sequence analysis will provide a clue in understanding the structure of the genome of Brassica rapa by analyzing the gene sequence, annotation and abundant repetitive DHA. The second stage involves sequencing of the genetically mapped seed BACs and identifying the overlapping BACs for complete genome sequencing. Currently, the second stage is comprises of process genetic anchoring using communal populations and maps to identify more than 1,000 seed BACs based on a BAC-to-BAC strategy. For the initial sequencing, 629 seed BACs corresponding to the minimum tiling path onto Arabidopsis genome were selected and fully sequenced. These BACs are now anchoring to the genetic map using the development of SSR markers. This information will be useful for identifying near BAC clones with the seed BAC on a genome map. From the BAC sequences, it is revealed that the Brassica rapa genome has extensive triplication of the DNA segment coupled with variable gene losses and rearrangements within the segments. This article introduces the current status and prospective of Korea Brassica Genome Project and the bioinformatics tools possessed in each national team. In the near future, data of the genome will contribute to improving Brassicas for their economic use as well as in understanding the evolutional process.

Analysis of rpoB Gene in Rifampin-Resistant M. Tuberculosis by Direct Sequencing and Line Probe Assay (염기서열결정과 Line Probe 분석법에 의한 Rifampin내성 결핵균의 rpoB 유전자 분석)

  • Lee, Min-Ki;Kim, Yun-Seong;Lee, Hyo-Jin;Cheon, Du-Su;Yun, Sang-Myung;Park, Sam-Seok;Kim, Cheol-Min;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.2
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    • pp.251-263
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    • 1997
  • Background : The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture. Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in $\beta$subunit of RNA polymerase is main mechanism of resistance. Method : In this study, rpoB gene for the $\beta$subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. Results : All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S531L) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not confirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513~515 were new identified mutations for the first time. Conclusion : Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.

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H2AX Directly Interacts with BRCA1 and BARD1 via its NLS and BRCT Domain Respectively in vitro (H2AX의 BRCA1 NLS domain과 BARD1 BRCT domain 각각과의 in vitro 상호 결합)

  • Bae, Seung-Hee;Lee, Sun-Mi;Kim, Su-Mi;Choe, Tae-Boo;Kim, Cha-Soon;Seong, Ki-Moon;Jin, Young-Woo;An, Sung-Kwan
    • KSBB Journal
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    • v.24 no.4
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    • pp.403-409
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    • 2009
  • H2AX, a crucial component of chromatin, is implicated in DNA repair, cell cycle check point and tumor suppression. The aim of this study was to identify direct binding partners of H2AX to regulate cellular responses to above mechanisms. Literature reviews and bioinformatical tools were attempted intensively to find binding partners of H2AX, which resulted in identifying two potential proteins, breast cancer-1 (BRCA1) and BRCA1-associated RING domain 1 (BARD1). Although it has been reported in vivo that BRCA1 co-localizes with H2AX at the site of DNA damage, their biochemical mechanism for H2AX were however only known that the complex monoubiquitinates histone monomers, including unphosphorylated H2AX in vitro. Therefore, it is important to know whether the complex directly interacts with H2AX, and also which regions of these are specifically mediated for the interaction. Using in vitro GST pull-down assay, we present here that BRCA1 and BARD1 directly bind to H2AX. Moreover, through combinational approaches of domain analysis, fragment clonings and in vitro binding assay, we revealed molecular details of the BRCA1-H2AX and BARD1-H2AX complex. These data provide the potential evidence that each of the BRCA1 nuclear localization signal (NLS) and BARD1 BRCA1 C-terminal (BRCT) repeat domain is the novel mediator of H2AX recognition.

Analysis of Nutritional Compounds and Antioxidant Effect of Freeze-Dried powder of the Honey Bee (Apis mellifera L.) Drone (Pupal stage) (서양종 꿀벌(Apis mellifera L.) 수벌번데기 동결건조 분말의 영양학적 성분 및 항산화 효과)

  • Kim, Jung-Eun;Kim, Do-Ik;Koo, Hui-Yeon;Kim, Hyeon-Jin;Kim, Seong-Yeon;Lee, Yoo-Beom;Kim, Ji-Soo;Kim, Ho-Hyuk;Moon, Jae-Hak;Choi, Yong-Soo
    • Korean journal of applied entomology
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    • v.59 no.3
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    • pp.265-275
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    • 2020
  • In this study, we analyzed the nutritional ingredients of drone pupae (16th to 20th instar old) to evaluate the value of bee products and provide basic data for product diversification, and the extracts prepared using these pupae were tested for physiological activity. According to the analysis of the general ingredients of the freeze-dried powder of these bee pupae, the moisture, crude protein, crude fat, and crude ash was 1.69 ± 0.07%, 48.52 ± 0.20%, 23.41 ± 0.14%, and 4.05 ± 0.02%, respectively. Vitamin C and vitamin E were 14.92 ± 0.52 mg/100 g and 6.06 ± 0.11 mg α-TE/100 g, respectively. Regarding minerals, the highest content of K (1349.13 ± 34.57 mg/100 g) and P (1323.55 ± 43.85 mg/100 g) was observed and Ca and Fe were 55.43 ± 1.51 mg/100 g and 5.49 ± 0.19 mg/100 g, respectively. The fatty acids of the water extracted freeze-dried pupae powder accounted for approximately 59.62 of saturated fatty acids and 40.38 of unsaturated fatty acids, and high-quality fatty acids such as palmitic acid (C16:0) was 35.49 ± 0.08 and oleic acid (C18:1, n-9) was 35.91 ± 0.22 (g/100 g total fatty acids). The total amino acid content was 38.99 ± 2.63 g/100 g and the free amino acid was a total of 5129.04 mg/100 g, of which 1257.68 mg/100 g was proline and 759.12 mg/100 g glutamic acid. The DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity of the drone pupae extract showed values of 0.8 for distilled water extract, 3.2 for 50% EtOH extract, 6.4 for 70% EtOH extract, and approximately 90% for 32 ㎍/mL for 100% EtOH extract. These results suggest that the main compound contributing to the antioxidant activity is a polar compound, and it is highly likely to be a low-molecular protein or a free amino acid. In conclusion, the honey bee drone pupa is excellent as a food resource and can be utilized as a new functional material for food and functional food.

Antioxidant Effect of Hot water and Ethanol extracts from Cheonnyuncho (Opuntia humifusa) on Reactive Oxygen Species (ROS) Production in 3T3-L1 Adipocytes (3T3-L1 지방세포내 ROS 생성에 대한 천년초 열수 및 에탄올 추출물의 항산화 효과)

  • Yoon, Bo-Ra;Lee, Young-Jun;Kim, Sun-Gu;Jang, Jung-Young;Lee, Hyo-Ku;Rhee, Seong-Kap;Hong, Hee-Do;Choi, Hyeon-Son;Lee, Boo-Yong;Lee, Ok-Hwan
    • Food Science and Preservation
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    • v.19 no.3
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    • pp.443-450
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    • 2012
  • Recently, NADPH oxidase 4 (NOX4)-mediated generation of intracellular reactive oxygen species (ROS) was proposed to accelerate adipogenesis of 3T3-L1 cell. We have previously shown that Cheonnyuncho (Opuntia humifusa) extract significantly inhibited adipocyte differentiation via downregulation of $PPAR{\gamma}$ (peroxisome proliferator-activated receptor gamma) gene expression. In this study, we focused on the molecular mechanism(s) of NOX4, G6PDH (glucose-6-phosphate dehydrogenase) and antioxidant enzymes in anti-oxidative activities of 3T3-L1 adipocytes. Our results indicate that Cheonnyuncho extracts markedly inhibits ROS production during adipogenesis in 3T3-L1 cells. Cheonnyuncho extracts suppressed the mRNA expression of the pro-oxidant enzyme such as NOX4 and the NADPH-producing G6PDH enzyme. In addition, treatment with Cheonnyuncho extract was found to upregulate mRNA levels of antioxidant enzymes such as Mn-SOD (manganese-superoxide dismutase), Cu/Zn-SOD (copper/zinc-SOD), glutathione peroxidase (GPx), glutathion reductase (GR), and catalase, all of which are important for endogenous antioxidant responses. These data suggest that Cheonnyuncho extract may be effective in preventing the rise of oxidative stress during adipocyte differentiation through mechanism(s) that involves direct down regulation of NOX4 and G6PDH gene expression or via upregulation of endogenous antioxidant responses.

Cytoprotective Effects of Schisandrin A against Hydrogen Peroxide-induced Oxidative Stress in SW1353 Human Chondrocytes (SW1353 인간 연골세포에서 산화적 스트레스에 대한 schisandrin A의 세포 보호 효과)

  • Jeong, Jin-Woo;Choi, Eun Ok;Kwon, Da Hye;Kim, Bum Hoi;Park, Dong Il;Hwang, Hye Jin;Kim, Byung Woo;Choi, Yung Hyun
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1070-1077
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    • 2017
  • Chondrocyte apoptosis induced by reactive oxygen species (ROS) plays an important role in the pathogenesis of osteoarthritis. Schisandrin A, a bioactive compound found in fruits of the Schisandra genus, has been reported to possess multiple pharmacological and therapeutic properties. Although several studies have described the antioxidant effects of analogues of schisandrin A, the underlying molecular mechanisms of this bioactive compound remain largely unresolved. The present study investigated the cytoprotective effect of schisandrin A against oxidative stress (hydrogen peroxide [$H_2O_2$]) in SW1353 human chondrocyte cells. The results showed that schisandrin A preconditioning significantly inhibited $H_2O_2-induced$ growth inhibition and apoptotic cell death by blocking the degradation of poly (ADP-ribose) polymerase proteins and down-regulating pro-caspase-3. These antiapoptotic effects of schisandrin A were associated with attenuation of mitochondrial dysfunction and normalization of expression changes of proapoptotic Bax and antiapoptotic Bcl-2 in $H_2O_2-stimulated$ SW1353 chondrocytes. Furthermore, schisandrin A effectively abrogated $H_2O_2-induced$ intracellular ROS accumulation and phosphorylation of histone H2AX at serine 139, a widely used marker of DNA damage. Thus, the present study demonstrates that schisandrin A provides protection against $H_2O_2-induced$ apoptosis and DNA damage in SW1353 chondrocytes, possibly by prevention of ROS generation. Collectively, our data indicate that schisandrin A has therapeutic potential in the treatment of oxidative disorders caused by overproduction of ROS.

Protective Effect of Canavalia gladiata on Gastric Inflammation Induced by Alcohol Treatment in Rats (알코올성 위염 동물 모델에서 작두콩 추출물의 보호 효과)

  • Kim, Ok Kyung;Nam, Da-Eun;You, Yanghee;Jun, Woojin;Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.5
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    • pp.690-696
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    • 2013
  • The objective of this study was to investigate the protective effect of extracts from Canavalia gladiata (CGE) on gastric inflammation induced by alcohol treatment in SD rats. Rats were divided into four groups: G1 (normal group), G2 (gastric inflammation induced by alcohol), G3 (gastric inflammation induced by alcohol with lansoprazole pretreatment), G4 (gastric inflammation induced by alcohol with 250 mg/kg b.w. CGE pretreatment), G5 (gastric inflammation induced by alcohol with 500 mg/kg b.w. CGE pretreatment). After the oral administration of 40% alcohol and samples for seven days, acute gastritis was induced with 70% alcohol and 0.15 M HCl. After 1 h of alcohol administration, the animals were sacrificed. Groups pretreated with lansoprazole or CGE showed an attenuation of gastric mucosal injury, including decreases in sub-epithelial loss, hemorrhages, and gastric juice secretion induced by administration of alcohol. The oral administration of CGE (500 mg/kg b.w.) significantly decreased the levels of TBARS. To examine molecular factors that regulate inflammation, the protein expression of NF-${\kappa}B$ and COX-2 were measured through immuno-histochemistry. Compared with the normal group (G1), the expression of NF-${\kappa}B$ and COX-2 were clearly increased in G2. COX-2 and NF-${\kappa}B$ were expressed even higher in groups pretreated with CGE compared to G2. In conclusion, our data show that Canavalia gladiata has inhibitory and protective effects on gastric inflammation induced by alcohol treatment in SD rats.

Differences of Tc-99m HMPAO SPECT Imaging in the Early Stage of Subcortical Vascular Dementia Compared with Alzheimer's Disease (초기 단계의 피질하 혈관성 치매와 알쯔하이머병에서 Tc-99m HMPAO SPECT 영상 소견 차이)

  • Park, Kyung-Won;Kang, Do-Young;Park, Min-Jeong;Cheon, Sang-Myung;Cha, Jae-Kwan;Kim, Sang-Ho;Kim, Jae-Woo
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.6
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    • pp.530-537
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    • 2007
  • Purpose: The aim of this study is to assess the specific patterns of regional cerebral blood flow (rCBF) in patients with the early stage of subcortical vascular dementia (SVaD) and Alzheimer's disease (AD) using Tc-99m HMPAO SPECT, and to compare the differences between the two conditions. Materials and Methods: Sixteen SVaD, 46 AD and 12 control subjects participated in this study. We included the patients with SVaD and AD according to NINCDS-ADRDA and NINDS-AIREN criteria. They were all matched for age, education and clinical dementia rating scores. Three groups were evaluated by Tc-99m HMPAO SPECT using statistical parametric mapping (SPM) for measuring rCBF. The SPECT data of patients with SVaD and AD were compared with those of normal control subjects and then compared with each other. Results: SPM analysis of the SPECT image showed significant perfusion deficits on the right temporal region and thalamus, left insula and superior temporal gyrus, both cingulate gyri and frontal subgyri in patients with SVaD and on the left supramarginal gyrus, superior temporal gyrus, postcentral gyrus and inferior parietal lobule, right fugiform gyrus and both cingulate gyri in AD compared with control subjects (uncorrected p<0.01). SVaD patients revealed significant hypoperfusion in the right parahippocampal gyrus with cingulated gyrus, left insula and both frontal subgyral regions compared with AD (uncorrected p<0.01). Conclusion: Our study shows characteristic and different pattern of perfusion deficits in patients with SVaD and AD, and these results may be helpful to discriminate the two conditions in the early stage of illness.

Solution Structure of 21-Residue Peptide (Asp 84-Leu 104), Functional Site derived from $p16^{INK4A}$ ($p16^{INK4A}$ 단백질 활성부위(Asp 84-Leu 104)의 용액상 구조)

  • Lee, Ho-Jin;Ahn, In-Ae;Ro, Seonggu;Choi, Young-Sang;Yoon, Chang No;Lee, Kang-Bong
    • Analytical Science and Technology
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    • v.13 no.4
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    • pp.494-503
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    • 2000
  • A 21-residue peptide corresponding to amino acids 84-104 of $p16^{INK4A}$, the tumor suppressor, has been synthesized and its structure was studied by Circular Dichroism, $^1H$ NMR spectroscopy and molecular modeling. A p16-derived peptide (84-104 amino acids) forming stable complex with CDK4 and CDK6 inhibits the ability of CDK4/6 to phosphorylate pRb in vitro, and blocks cell-cycle progression through G1/S phase as shown in the function of the full-length p16. Its NMR spectral data including NOEs, $^3J_{NH-H{\alpha}}$ coupling constants, $C_{\alpha}H$ chemical shift, the average amplitude of amide chemical shift oscillation and temperature coefficients indicate that the secondary structure of a p16-derived peptide is similar to that of the same region of full-length p16, which consists of helix-turn-helix structure. The 3-D distance geometry structure based on NOE-hased distance and torsion angle restraints is characterized by ${\gamma}$-turn conformation between residues $Gly^{89}-Leu^{91}$(${\varphi}_{i+1}=-79.8^{\circ}$, ${\varphi}_{i+1}=60.2^{\circ}$) as evidenced in a single crystal structure for the corresponding region of p18 or p19, but is undefined at both the N and C termini. This compact and rigid ${\gamma}$-turn region is considered to stabilize the structure of p16-derived peptide and serve as a site recognizing cyelin dependent kinase, and this well-defined ${\gamma}$-turn structure could be utilized for the design of anti-cancer drug candidates.

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Development of Dynamic Kidney Phantom System and its Evaluation of Usability of Application in Nuclear Medicine (핵의학 동적 신장팬텀시스템 개발 적용의 유용성 평가)

  • Park, Hoon-Hee;Lee, Juyoung;Kim, Sang-Wook;Lyu, Kwang Yeul;Jin, Gye Hwan
    • Journal of radiological science and technology
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    • v.36 no.1
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    • pp.49-55
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    • 2013
  • Currently, commercially available phantom can reproduce and evaluate only a static situation, the study is incomplete research on phantom and system which is can confirmed functional situation in the kidney by time through dynamic phantom and blood flow velocity, various difference according to the amount of radioactive. Therefore, through this study, it has produced the dynamic kidney phantom to reproduce images through the dynamic flow of the kidney, it desires to evaluate the usefulness of nuclear medicine imaging. The production of the kidney phantom was fabricated based on the normal adult kidney, in order to reproduce the dynamic situation based on the fabricated kidney phantom, in this study, it was applied the volume pump that can adjust the speed of blood flow, so it can be integrated continuously radioactive isotopes in the kidney by using $^{99m}Tc$-pertechnate. Used the radioactive isotope was supplied through the two pump. It was confirmed the changes according to the infusion rate, radioactive isotopes and the different injection speeds on the left and right, analysis of the acquired images was done by drawn five times ROI in order to check the reproducibility of each on the front and rear of the kidney and bladder. Depending on the speed of injection, radioisotope was a lot of integrated and emissions up when adjusting the pressure of the pump as 30 stroke, it was the least integrated and emissions up when adjusting as 40 stroke. The integration of the left & right kidney was not reached in the amount of the highest when adjusting as 10 stroke. In the changes according to the amount of the radioactive isotope, 0.6 mCi(22.2 MBq), 0.8 mCi (29.6 MBq)was showed up similar tendency but, in the result of the different injection 0.8 mCi, it was showed up counts close to double of 0.6 mCi. In the result of the differently injection speed of the left & right kidney, as a result of different conditions that injection speed was 20 stroke through left kidney phantom, the injection speed was 30 stroke through right kidney phantom, it was enough difference in the resulting image can be easily distinguished with the naked eye. Through this study, the results showed that the dynamic kidney phantom system is able to similarly reproduce renogram in the actual clinical practice. Especially, the depicted over time for the flow to be excreted through the kidney into the bladder was adequately reproduce, it is expected to be utilized as basic data to check the quality of the dynamic images. In addition, it is considered to help in the field of functional imaging and quality control.