• Title/Summary/Keyword: stem region

Search Result 320, Processing Time 0.027 seconds

Mutational Analysis of an Essential RNA Stem-loop Structure in a Minimal RNA Substrate Specifically Cleaved by Leishmania RNA Virus 1-4 (LRV1-4) Capsid Endoribonuclease

  • Ro, Youngtae;Patterson, Jean L.
    • Journal of Microbiology
    • /
    • v.41 no.3
    • /
    • pp.239-247
    • /
    • 2003
  • The LRV1-4 capsid protein possesses an endoribonuclease activity that is responsible for the single site-specific cleavage in the 5' untranslated region (UTR) of its own viral RNA genome and the formation of a conserved stem-loop structure (stem-loop IV) in the UTR is essential for the accurate RNA cleavage by the capsid protein. To delineate the nucleotide sequences, which are essential for the correct formation of the stem-loop structure for the accurate RNA cleavage by the viral capsid protein, a wildtype minimal RNA transcript (RNA 5' 249-342) and several synthetic RNA transcripts encoding point-mutations in the stem-loop region were generated in an in vitro transcription system, and used as substrates for the RNA cleavage assay and RNase mapping studies. When the RNA 5' 249-342 transcript was subjected to RNase T1 and A mapping studies, the results showed that the predicted RNA secondary structure in the stem-loop region using FOLD analysis only existed in the presence of Mg$\^$2+/ ions, suggesting that the metal ion stabilizes the stem-loop structure of the substrate RNA in solution. When point-mutated RNA substrates were used in the RNA cleavage assay and RNase T1 mapping study, the specific nucleotide sequences in the stem-loop region were not required for the accurate RNA cleavage by the viral capsid protein, but the formation of a stem-loop like structure in a region (nucleotides from 267 to 287) stabilized by Mg$\^$2+/ ions was critical for the accurate RNA cleavage. The RNase T1 mapping and EMSA studies revealed that the Ca$\^$2+/ and Mn$\^$2+/ ions, among the reagents tested, could change the mobility of the substrate RNA 5' 249-342 on a gel similarly to that of Mg$\^$2+/ ions, but only Ca$\^$2+/ ions identically showed the stabilizing effect of Mg$\^$2+/ ions on the stem-loop structure, suggesting that binding of the metal ions (Mg$\^$2+/ or Ca$\^$2+/) onto the RNA substrate in solution causes change and stabilization of the RNA stem-loop structure, and only the substrate RNA with a rigid stem-loop structure in the essential region can be accurately cleaved by the LRV1-4 viral capsid protein.

Primary Intracranial Squamous Cell Carcinoma in the Brain Stem with a Cerebellopontine Angle Epidermoid Cyst

  • Kim, Min-Su;Kim, Oh-Lyong
    • Journal of Korean Neurosurgical Society
    • /
    • v.44 no.6
    • /
    • pp.401-404
    • /
    • 2008
  • Primary intracranial squamous cell carcinoma is extremely rare, with most cases arising from a preexisting benign epidermoid cyst. We report a rare case of primary intracranial squamous cell carcinoma in the brain stem with a cerebellopontine angle (CPA) epidermoid cyst. A 72-year-old female suffered from progressive left hemiparesis, difficulty in swallowing, and right hemifacial numbness. Diffusion-weighted magnetic resonance imaging revealed a high signal intensity (SI) lesion in the CPA region and an intra-axially ring-enhanced cystic mass in the right brain stem with low SI. Whole-body positron emission tomography showed no evidence of metastatic disease. The histological findings revealed a typical epidermoid cyst in the CPA region and a squamous cell carcinoma in the brain stem. We speculate that the squamous cell carcinoma may have been developed due to a chronic inflammatory response by the adjacent epidermoid cyst. The patient underwent a surgical resection and radiotherapy. After 12 months, she had no evidence of recurrence.

Development of Local Stem Volume Table for Pinus densiflora S. et Z. Using Tree Stem Taper Model (수간곡선 모델을 이용한 소나무의 지방별 수간재적표 개발)

  • Kang, Jin-Taek;Son, Yeong-Mo;Kim, So-Won;Lee, Sun-Jeoung;Park, Hyun
    • Korean Journal of Agricultural and Forest Meteorology
    • /
    • v.16 no.4
    • /
    • pp.327-335
    • /
    • 2014
  • Current volume tables might underestimate or overestimate the volumes of individual trees in a specific region because the tables were made using the data from broad regions within South Korea. Therefore, to solve this problem, this study was conducted to develop local stem volume tables reflecting the local growth pattern and properties using stem taper equations in the regions of Hongcheon and Yeongju. We developed the local stem volume table for Pinus densiflora, which is the widely planted species in South Korea. To derive the most suitable taper equation for estimating the stem volume of region, three models of Max & Burkhart, Kozak and Parresol et al. were applied and their fitness were statistically analyzed by using the Fitness Index, Bias, and Standard Error of Bias. The result showed that there is a significant difference among the three models, and the Fitness Index of the Kozak model was highest compared to the other models. Therefore, the Kozak model was chosen for generating stem taper equation and stem volume tables for P. densiflora. The result from the developed stem volume tables of each region was compared to the current stem volume tables with driven by the data of tree growth obtained throughout the nation. The result showed that there is a significant difference (0.000< ${\alpha}=0.05$) in two regions, Hongcheon and Yeongju, and also there is a significant difference (0.000< ${\alpha}=0.05$) between the two regions.

Stress Analysis of Femoral Stems on Non-Cemented Total Hip Replacement - A Three-Dimensional Finite Element Analysis -

  • Kim, Sung-Kon;Chae, Soo-Won;Jeong, Jung-Hwan
    • Proceedings of the KOSOMBE Conference
    • /
    • v.1997 no.11
    • /
    • pp.263-266
    • /
    • 1997
  • Three dimensional numerical model based on the finite element method(FEM) were developed to predict the mechanical behavior of hip implants. The purpose of this study is to investigate the stress distribution of two types of cementless total hip replacement femoral component -a straight stem and a curved stem, and to compare their effect on the stress shielding between two types by three dimensional finite element method. The authors analyzed von Mises stress in the cortex & stem and compared the stress between the straight and the curved stem. In comparison of stresses between two different design of femoral stem, there was 25% more decrease of stress in straight stem than curved stem in the medial cortex at proximal region. The straight stem had consistently much lower stresses than the curved stem throughout the whole medial cortex with maximum 70% reduction of stress. However, there was little change in stress between nature and 2 implanted femur throughout the lateral cortex. Stress of femoral stem was much higher in the straight stem than the curved stem up to 60%. The straight stem had more chance of stress shielding and a risk of fatigue fracture of the stem compared with the curved stem in noncement hip arthroplasty. In design of femoral stem still we have to consider to develop design to distribute more even stress on the proximal medial cortex.

  • PDF

A Study for Improvement of the Femoral Stem Type using the Finite Element Analysis (유한요소법을 이용한 인공고관절 주대형태의 개선에 대한 연구)

  • 윤경렬;원예연;이수훈
    • Korean Journal of Computational Design and Engineering
    • /
    • v.5 no.2
    • /
    • pp.122-126
    • /
    • 2000
  • A major mechanical problem with total hip replacement is the loosening of the femoral component. The loss of proximal support, with firm fixation distally, has been thought to be a major caused of fatigue failure of femoral stems. While many causes have been proposed, the most frequently suggested cause of the calcar resorption is the disuse atrophy of the cortex of the calcar due to the stress shielding of the proximal bone by the metal femoral stem. In this research, the new-designed stem(modified collar stem) was considered which made a hole inside stem and had a 3 mm thickness. Using the 3-dimensional finite element methods, the common collar stem and the modified colla stem was modeled and analysed. Also, the two models was compared. The results showed that the modified collar stem decreased the stress-shielding and it made a effective load transfer at the entire femoral region.

  • PDF

Distributions of HLA Microsatellite Markers and the Linkage Disequilibria between HLA and Microsatellites in Koreans (한국인에서 HLA 유전자 부위 내 Microsatellite 표지자의 분포와 HLA 대립유전자의 유전적 연관성)

  • Jang, Jung-Pil;Choi, Eun-Jeong;Yoon, Ho-Yeul;Choi, Hee-Baeg;Kim, Hee-Je;Cho, Byung-Sik;Min, Woo-Sung;Lee, Jong-Wook;Kim, Chun-Choo;Kim, Tai-Gyu
    • IMMUNE NETWORK
    • /
    • v.7 no.3
    • /
    • pp.149-157
    • /
    • 2007
  • Background: The microsatellites within human leukocyte antigen (HLA) region show considerable polymorphism and strong linkage disequilibrium (LD) with HLA alleles. These microsatellites have been used for genetic analysis including disease mapping to understand susceptibility to autoimmune and infectious diseases. Also, use of microsatellites has recently been proposed as an approach for identifying non-HLA markers within the HLA region that could function as transplantation determinants and for the selection of potential donors for transplantation. Methods: To analyse the frequency of five microsatellites in the Korean population, genotyping for polymorphisms at five microsatellites markers (BAT2, MIB, DQCAR, D6S105 and TNFd) within HLA region was performed on 143 healthy Korean controls. Results: The most frequent genotype shown in healthy Korean controls were BAT2 8 (153 bp, 42.7%), MIB 1 (326 bp, 40.6%), DQCAR 3 (188 bp, 38.5%), D6S105 7 (126 bp, 58.0%) and TNFd 3 (128 bp, 58.0%). And common two-loci haplotypes were found as MIB 1-HLA-B*62 (HF: 10.6%), MIB 6-HLA-B*44 (HF: 7.8%), DQCAR 3-HLA-DRB1*13 (HF: 8.5%), TNFd 5-HLA-B*62 (HF: 7.8%) and D6S105 7-HLA-A*02 (HF: 16.2%). Conclusion: These data might provide useful information on the microsatellites markers with HLA region in Korean population and be helpful in further defining the clinical impact of these microsatellites.

The expression and functional roles of microRNAs in stem cell differentiation

  • Shim, Jiwon;Nam, Jin-Wu
    • BMB Reports
    • /
    • v.49 no.1
    • /
    • pp.3-10
    • /
    • 2016
  • microRNAs (miRNAs) are key regulators of cell state transition and retention during stem cell proliferation and differentiation by post-transcriptionally downregulating hundreds of conserved target genes via seed-pairing in their 3' untranslated region. In embryonic and adult stem cells, dozens of miRNAs that elaborately control stem cell processes by modulating the transcriptomic context therein have been identified. Some miRNAs accelerate the change of cell state into progenitor cell lineages—such as myoblast, myeloid or lymphoid progenitors, and neuro precursor stem cells—and other miRNAs decelerate the change but induce proliferative activity, resulting in cell state retention. This cell state choice can be controlled by endogenously or exogenously changing miRNA levels or by including or excluding target sites. This control of miRNA-mediated gene regulation could improve our understanding of stem cell biology and facilitate their development as therapeutic tools. [BMB Reports 2016; 49(1): 3-10]

The SL1 Stem-Loop Structure at the 5′-End of Potato virus X RNA Is Required for Efficient Binding to Host Proteins and forViral Infectivity

  • Kwon, Sun-Jung;Kim, Kook-Hyung
    • Molecules and Cells
    • /
    • v.21 no.1
    • /
    • pp.63-75
    • /
    • 2006
  • The 5′-region of Potato virus X (PVX) RNA, which contains an AC-rich, single-stranded region and stem-loop structure 1 (SL1), affects RNA replication and assembly. Using Systemic Evolution of Ligands by EXponential enrichment (SELEX) and the electrophoretic mobility shift assay, we demonstrate that SL1 interacts specifically with tobacco protoplast protein extracts (S100). The 36 nucleotides that correspond to the top region of SL1, which comprises stem C, loop C, stem D, and the tetra loop (TL), were randomized and bound to the S100. Remarkably, the wild-type (wt) sequence was selected in the second round, and the number of wt sequences increased as selection proceeded. All of the selected clones from the fifth round contained the wt sequence. Secondary structure predictions (mFOLD) of the recovered sequences revealed relatively stable stem-loop structures that resembled SL1, although the nucleotide sequences therein were different. Moreover, many of the clones selected in the fourth round conserved the TL and C-C mismatch, which suggests the importance of these elements in host protein binding. The SELEX clone that closely resembled the wt SL1 structure with the TL and C-C mismatch was able to replicate and cause systemic symptoms in plants, while most of the other winners replicated poorly only on inoculated leaves. The RNA replication level on protoplasts was also similarly affected. Taken together, these results indicate that the SL1 of PVX interacts with host protein(s) that play important roles related to virus replication.

Characterization of the Nanog 5'-flanking Region in Bovine

  • Choi, Don-Ho;Kim, Duk-Jung;Song, Ki-Duk;Park, Hwan-Hee;Ko, Tae Hyun;Pyao, Yuliya;Chung, Ku-Min;Cha, Seok Ho;Sin, Young-Su;Kim, Nam-Hyung;Lee, Woon-Kyu
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.29 no.10
    • /
    • pp.1383-1391
    • /
    • 2016
  • Bovine embryonic stem cells have potential for use in research, such as transgenic cattle generation and the study of developmental gene regulation. The Nanog may play a critical role in maintenance of the undifferentiated state of embryonic stem cells in the bovine, as in murine and human. Nevertheless, efforts to study the bovine Nanog for pluripotency-maintaining factors have been insufficient. In this study, in order to understand the mechanisms of transcriptional regulation of the bovine Nanog, the 5'-flanking region of the Nanog was isolated from ear cells of Hanwoo. Results of transient transfection using a luciferase reporter gene under the control of serially deleted 5'-flanking sequences revealed that the -134 to -19 region contained the positive regulatory sequences for the transcription of the bovine Nanog. Results from mutagenesis studies demonstrated that the Sp1-binding site that is located in the proximal promoter region plays an important role in transcriptional activity of the bovine Nanog promoter. The electrophoretic mobility shift assay with the Sp1 specific antibody confirmed the specific binding of Sp1 transcription factor to this site. In addition, significant inhibition of Nanog promoter activity by the Sp1 mutant was observed in murine embryonic stem cells. Furthermore, chromatin-immunoprecipitation assay with the Sp1 specific antibody confirmed the specific binding of Sp1 transcription factor to this site. These results suggest that Sp1 is an essential regulatory factor for bovine Nanog transcriptional activity.

Secondary Structure of the Ribosomal Internal Transcribed Spacer (ITS) Region of Hypsizygus marmoreus (느티만가닥버섯의 ITS (internal transcribed spacer) 영역의 2차구조 분석)

  • Woo, Ju-Ri;Yoon, Hyeokjun;You, Young-Hyun;Lee, Chang-Yun;Kong, Won-Sik;Kim, Jong-Guk
    • Journal of Life Science
    • /
    • v.23 no.10
    • /
    • pp.1260-1266
    • /
    • 2013
  • The ribosomal DNA (rDNA) clusters of Hypsizygus marmoreus 3-10 and H. marmoreus 1-1 were analyzed in this study. The small subunit (SSU) and intergenic spacer 2 (IGS 2) was partially sequenced. The internal transcribed spacer 1 (ITS 1), 5.8S, internal transcribed spacer 2 (ITS 2), large subunit (LSU), intergenic spacer 1 (IGS 1), and 5S were completely sequenced. The rDNA clusters of H. marmoreus 3-10 and H. marmoreus 1-1 were 7,049 bp in length. The sequence of SSU rDNA, which corresponded to 18S rDNA, was 1,796 bp in length, and the sequence of LSU rDNA, which corresponded to 28S rDNA, was 3,348 bp in length. The ITS region that variable region and IGS region that non-transcribed spacer was 462 bp and 1,290 bp in length. The sequence of 5.8S rDNA and 5S rDNA was 153 bp and 43 bp in length, respectively. The 17 bp of the rDNA cluster in the H. marmoreus 3-10 strain was different to that in the H. marmoreus 1-1 strain, with 2 bp in the SSU, 3 bp in the ITS, 9 bp in the LSU, and 3 bp in the IGS. The analysis of the secondary structure revealed that the ITS regions of H. marmoreus 3-10 and H. marmoreus 1-1 have five stem-loop structures. Interestingly, among these structures, one different nucleotide sequence resulted in a different secondary structure in stem-loop V.