• Title/Summary/Keyword: RNA 1 structure

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Expression of Luteinizing Hormone (LH) Subunit Genes in the Rat Ovary (흰쥐 난소에서의 Luteinizing Hormone (LH) Subunit 유전자 발현)

  • Lee, Sung-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.199-205
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    • 1998
  • The present study was performed to analyze the expression of LH genes in the rat ovary. Expression of LH subunit genes in the rat ovary was demonstrated by amplification of ovarian RNA by RT-PCR. The ovarian $LH_\beta$ transcripts contained at least two parts of the published cDNA structure, the pituitary exons 1, 2 and 3 and the part of testicular ex on 1 in the major trancripts form in rat testis. Using RIA, significant amount of LH-like molecules were detected in crude ovarian extracts, and the competition curves with increasing amount of tissue extracts were parallel with those of standard peptide, indicating that the ovarian immunoreactive LH-like material is similar to authentic pituitary LH molecule. The administration of PMSG to immature rats resulted in a sharp decrease of the ovarian LH contents after 24 h post-injection. In conclusion, these findings demonstrate that genes for LH subunits are expressed in the rat ovary, and suggest that LH can playa central role in regulation of female reproduction with both endocrine (by pituitary LH) and auto- and/or para-crine (by ovarian LH) manner.

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Characterization of a Monosaccharide Transporter mstB Isolated as a Downstream Gene of MsnA in Aspergillus nidulans (Aspergillus nidulans에서 MsnA 하위 유전자로 선별된 단당류 수송자 mstB의 기능 분석)

  • Jeon, Mee-Hyang;Chae, Suhn-Kee
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.281-288
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    • 2011
  • To screen downstream genes of Aspergillus nidulans MsnA showing amino acid sequence similarity to the zinc finger region of Msn2/4 stress response transcription factors in Saccharomyces cerevisiae, differentially expressed genes (DEG) in MsnA overexpressed or msnA null mutant strains compared to wild type have been isolated. The cognate gene IDs were identified by DNA sequencing of the selected DEGs. Among those, DEG6 was known as mstB encoding a putative monosaccharide transporter. Expression level of mstB mRNA was increased in MsnA overproducing strains and MsnA bound directly to the promoter region of mstB in vitro. MstB containing twelve transmembrane domains exhibited 80% of amino acid sequence identities to A. niger MstA a high-affinity monosaccharide transporter. A null mutant of mstB was phenotypically undistinguishable to wild type. On the other hand, forced overexpression of MstB caused the increased formation of sexual structure cleistothecia in 0.1% glucose condition where wild type showed almost no cleistothecia. This result implies that mstB is involved in transport of monosaccharide required for sexual differentiation.

Evaluation of Construction Operation and Design Properties of CLSM for Corrugated Pipe in Underground (파형강관을 이용한 지하매설물용 뒤채움재 설계 및 시공성 평가)

  • Lee Kwan-Ho;Park Jae-Heon
    • International Journal of Highway Engineering
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    • v.8 no.2 s.28
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    • pp.63-74
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    • 2006
  • During the construction of circular underground pipe, the non-proper compaction along the pipe and the decrease of compaction efficiency have been the main problems to induce the failure of underground pipe or facility. The use of CLSM (controlled low strength materials) should be one of the possible applications to overcome those problems. In this research, the full-scaled field test and the numeric analysis using PENTAGON-3D FEM program were carried out for three different cases on the change of backfill materials, including the common sand, the soil from construction site, and the CLSM. From the full-scaled test in field, the use of in-situ CLSM as backfill materials reduced the vertical and lateral deformation of the pipe, as well as the deformation of the ground surface. The main reason for reducing the deformation would be the characteristics of the CLSM, especially self-leveling and self-hardening properties. The measured earth pressure at the surround of the corrugated pipe using the CLSM backfills was the smaller than the other cases, and the absolute value was almost zero. Judging from the full-scaled field test and FEM analysis, the use of CLSM as backfill materials should be one of the best choices reducing the failure of the underground pipes.

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Copy Number Deletion Has Little Impact on Gene Expression Levels in Racehorses

  • Park, Kyung-Do;Kim, Hyeongmin;Hwang, Jae Yeon;Lee, Chang-Kyu;Do, Kyoung-Tag;Kim, Heui-Soo;Yang, Young-Mok;Kwon, Young-Jun;Kim, Jaemin;Kim, Hyeon Jeong;Song, Ki-Duk;Oh, Jae-Don;Kim, Heebal;Cho, Byung-Wook;Cho, Seoae;Lee, Hak-Kyo
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.9
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    • pp.1345-1354
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    • 2014
  • Copy number variations (CNVs), important genetic factors for study of human diseases, may have as large of an effect on phenotype as do single nucleotide polymorphisms. Indeed, it is widely accepted that CNVs are associated with differential disease susceptibility. However, the relationships between CNVs and gene expression have not been characterized in the horse. In this study, we investigated the effects of copy number deletion in the blood and muscle transcriptomes of Thoroughbred racing horses. We identified a total of 1,246 CNVs of deletion polymorphisms using DNA re-sequencing data from 18 Thoroughbred racing horses. To discover the tendencies between CNV status and gene expression levels, we extracted CNVs of four Thoroughbred racing horses of which RNA sequencing was available. We found that 252 pairs of CNVs and genes were associated in the four horse samples. We did not observe a clear and consistent relationship between the deletion status of CNVs and gene expression levels before and after exercise in blood and muscle. However, we found some pairs of CNVs and associated genes that indicated relationships with gene expression levels: a positive relationship with genes responsible for membrane structure or cytoskeleton and a negative relationship with genes involved in disease. This study will lead to conceptual advances in understanding the relationship between CNVs and global gene expression in the horse.

ANXA2 Regulates the Behavior of SGC-7901 Cells

  • Sun, Meng-Yao;Xing, Rui-Huan;Gao, Xiao-Jie;Yu, Xiang;He, Hui-Min;Gao, Ning;Shi, Hong-Yan;Hu, Yan-Yan;Wang, Qi-Xuan;Xu, Jin-Hui;Hou, Ying-Chun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6007-6012
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    • 2013
  • ANXA2, a member of the annexin family, is overexpressed and plays important roles in tumor development. However, the significance of ANXA2 expression in gastric carcinoma has not been clarified.To elucidate its roles in growth of gastric cancer, ANXA2 expression in SGC-7901 cells was inhibited with a designated siRNA, then cell proliferation, cell cycling, apoptosis and motility were determined by MTT assay, flow cytometry, Hoechst 33342 staining and wound healing assay, respectively. To further assess the behavior of ANXA2 deleted SGC-7901 cells, changes of microstructures were observed under fluorescence microscopy, laser scanning confocal microscopy and electron microscopy. We found that inhibition of ANXA2 expression caused cell proliferation to decrease significantly with G1 arrest, motility to be reduced with changes in pseudopodia/filopodia structure and F-actin and ${\beta}$-tubulin expression, and apoptosis to be enhanced albeit without significance. At the same time, ANXA2 deletion resulted in fewer pseudopodia/filopodia, non-stained areas were increased, contact inhibition among cells reappeared, and expression of F-actin and ${\beta}$-tubulin was decreased, with induction of polymerized disassembled forms. Taken together, these data suggest that ANXA2 overexpression is important to maintain the malignancy of cancer cells, and this member of the annexin family has potential to be considered as a target for the gene therapy of gastric carcinoma.

Genetic characterisation of PPARG, CEBPA and RXRA, and their influence on meat quality traits in cattle

  • Goszczynski, Daniel Estanislao;Mazzucco, Juliana Papaleo;Ripoli, Maria Veronica;Villarreal, Edgardo Leopoldo;Rogberg-Munoz, Andres;Mezzadra, Carlos Alberto;Melucci, Lilia Magdalena;Giovambattista, Guillermo
    • Journal of Animal Science and Technology
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    • v.58 no.4
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    • pp.14.1-14.9
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    • 2016
  • Background: Peroxisome proliferator-activated receptor gamma (PPARG), CCAAT/enhancer binding protein alpha (CEBPA) and retinoid X receptor alpha (RXRA) are nuclear transcription factors that play important roles in regulation of adipogenesis and fat deposition. The objectives of this study were to characterise the variability of these three candidate genes in a mixed sample panel composed of several cattle breeds with different meat quality, validate single nucleotide polymorphisms (SNPs) in a local crossbred population (Angus - Hereford - Limousin) and evaluate their effects on meat quality traits (backfat thickness, intramuscular fat content and fatty acid composition), supporting the association tests with bioinformatic predictive studies. Results: Globally, nine SNPs were detected in the PPARG and CEBPA genes within our mixed panel, including a novel SNP in the latter. Three of these nine, along with seven other SNPs selected from the Single Nucleotide Polymorphism database (SNPdb), including SNPs in the RXRA gene, were validated in the crossbred population (N = 260). After validation, five of these SNPs were evaluated for genotype effects on fatty acid content and composition. Significant effects were observed on backfat thickness and different fatty acid contents (P < 0.05). Some of these SNPs caused slight differences in mRNA structure stability and/or putative binding sites for proteins. Conclusions: PPARG and CEBPA showed low to moderate variability in our sample panel. Variations in these genes, along with RXRA, may explain part of the genetic variation in fat content and composition. Our results may contribute to knowledge about genetic variation in meat quality traits in cattle and should be evaluated in larger independent populations.

Phylogenetic Diversity and Community Structure of Microbiome Isolated from Sargassum Horneri off the Jeju Island Coast (제주 연안의 괭생이모자반(Sargassum horneri)에서 분리된 세균의 계통학적 다양성 및 군집 구조 분석)

  • Moon, Kyung-Mi;Park, So-Hyun;Heo, Moon-Soo
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1179-1185
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    • 2018
  • Recently, Sargassum horneri, the marine weed inhabiting the shoreline, beach, and littoral sea area, has caused serious damage to intensive aquaculture farms particularly those around Jeju Island, South Korea. The purpose of this study was to investigate the diversity of microorganisms in Sargassum horneri and to provide basic data on ecological problems by identifying microbial functions. A total of 88 isolates were identified by 16S rRNA sequencing. Proteobacteria was the dominant phylum accounting for 88%, including class ${\alpha}-proteobacteria$, six genera, and ten species. The dominating genus, Pseudobacter, accounted for 40% in Pseudorhodobacter, 20% in Paracoccus, and the remaining at 10% each were Rhizobium, Albirhodobacter, Skermanella, and Novosphingobium. Class ${\beta}-proteobactera$ included five genera and ten species. Genus Hydrogenophaga accounted for 50%, while genus Azoarcus accounted for 20%, and the remaining Oxalicibacterium, Duganella, and Xenophilus were 10% each. Class ${\gamma}-proteobacteria$ with 13 genera and 57 species, accounted for 74% in phylum Proteobacteria, 23% in Shewanella, 19% in Cobetia, 12% in Pseudomonas, 4% each in Vibrio and Serratia, and 2% each in Rheinheimera, Raoultella, Pantoea, Acinetobacter, Moraxella, and Psychrobacter genera. In addition, Actinobacteria with two species of Nocardioides genera accounted for 50%, and Bacteroidetes accounted for 33%, with three genera and five species that included Lacihabitans and Mariniflexile. The remaining Dyadobacter, Cellulophaga, and Ferruginibacter genera each accounted for 11%.

Regulation of cementoblast differentiation and mineralization using conditioned media of odontoblast (상아모세포의 조건배지를 이용한 백악모세포의 분화와 석회화 조절)

  • Moon, Sang-Won;Kim, Hye-Sun;Song, Hyun-Jung;Choi, Hong-Kyu;Park, Jong-Tae;Kim, Heung-Joong;Jang, Hyun-Seon;Park, Joo-Cheol
    • Journal of Periodontal and Implant Science
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    • v.36 no.2
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    • pp.385-396
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    • 2006
  • For the regeneration of periodontal tissues, the microenvironment for new attachment of connective tissue fibers should be provided, At this point of view, cementum formation in root surface plays a key role for this new attachment. This study was performed to figure out which factor promotes differentiation of cementoblast Considering anatomical structure of tooth, we selected the cells which may affect the differentiation of cementoblast - Ameloblast, OD11&MDPC23 for odontoblasts, NIH3T3 for fibroblsts and MG63 for osteoblasts. And OCCM30 was selected for cementoblast cell line. Then, the cell lines were cultured respectively and transferred the conditioned media to OCCM30. To evaluate the result, Alizarin red S stain was proceeded for evaluation of mineralization. The subjected mRNA genes are bone sialoprotein(BSP), alkaline phosphate(ALP) , osteocalcin(OC), type I collagen(Col I), osteonectin(SPARC ; secreted protein acidic and rich in cysteine). Expression of the gene were analysed by RT-PCR, The results were as follows: 1. For alizarin red S staining, control OCCM30 didn't show any mineralized red nodules until 14 days. But red nodules started to appear from about 4 days in MDPC-OCCM30 & OD11-OCCM30. 2. For results of RT-PCR, ESP mRNAs of control-OCCM30 and others were expressed from 14 days, but in MDPC23-OCCM30 & OD11-OCCM30 from 4 days. Like this, the gene expression of MDPC23-OCCM30 & OD11-OCCM30 were detected much earlier than others. 3. For confirmation of odontoblast effect on cementoblast, conditioned media of osteoblasts(MG63) which is mineralized by producing matrix vesicles didn't affect on the mineralized nodule formation of cementoblasts(OCCM30). This suggest the possibility that cementoblast mineralization is regulated by specific factor in dentin matrix protein rather than matrix vesicles. Therefore, we proved that the dentin/odontoblast promotes differentiation/mineralization of cementoblasts. This new approach might hole promise as diverse possibilities for the regeneration of tissues after periodontal disease.

Stage specific transcriptome analysis of liver tissue from a crossbred Korean Native Pig (KNP × Yorkshire)

  • Kumar, Himansu;Srikanth, Krishnamoorthy;Park, Woncheol;Lee, Kyung-Tai;Choi, Bong-Hwan;Kim, Jun-Mo;Lim, Dajeong;Park, Jong-Eun
    • Journal of Biomedical and Translational Research
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    • v.19 no.4
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    • pp.116-124
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    • 2018
  • Korean Native Pig (KNP) has a uniform black coat color, excellent meat quality, white colored fat, solid fat structure and good marbling. However, its growth performance is low, while the western origin Yorkshire pig has high growth performance. To take advantage of the unique performance of the two pig breeds, we raised crossbreeds (KNP ${\times}$ Yorkshire to make use of the heterotic effect. We then analyzed the liver transcriptome as it plays an important role in fat metabolism. We sampled at two stages: 10 weeks and at 26 weeks. The stages were chosen to correspond to the change in feeding system. A total of 16 pigs (8 from each stage) were sampled and RNA sequencing was performed. The reads were mapped to the reference genome and differential expression analysis was performed with edgeR package. A total of 324 genes were found to be significantly differentially expressed (${\left|log2FC\right|}$ > 1 & q < 0.01), out of which 180 genes were up-regulated and 144 genes were down-regulated. Principal Component Analysis (PCA) showed that the samples clustered according to stages. Functional annotation of significant DEGs (differentially expressed genes) showed that GO terms such as DNA replication, cell division, protein phosphorylation, regulation of signal transduction by p53 class mediator, ribosome, focal adhesion, DNA helicase activity, protein kinase activity etc. were enriched. KEGG pathway analysis showed that the DEGs functioned in cell cycle, Ras signaling pathway, p53 signaling pathway, MAPK signaling pathway etc. Twenty-nine transcripts were also part of the DEGs, these were predominantly Cys2His2-like fold group (C2H2) family of zinc fingers. A protein-protein interaction (PPI) network analysis showed that there were three highly interconnected clusters, suggesting an enrichment of genes with similar biological function. This study presents the first report of liver tissue specific gene regulation in a cross-bred Korean pig.

Anti-inflammatory Metabolites of Agrimonia pilosa Ledeb. and Their Mechanism

  • Park, Mi Jin;Ryu, Da Hye;Cho, Jwa Yeoung;Kang, Young-Hwa
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.13-13
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    • 2018
  • The anti-inflammatory (INF) compounds (1-15) were isolated from Agrimonia pilosa Ledeb. (APL) by activity-guided isolation technique. The isolated compounds (1-15) were identified as quercetin-7-O-rhanmoside (1), apigenin-7-O-glycoside (2), kaempferol-7-O-glycoside (3), apigenin-7-O-[6"-(butyl)-glycoside] (4), querceitn (5), kaempferol (6), apigenin (7), apigenin-7-O-[6"-(pentyl)-glycoside] (8), agrimonolide (9), agrimonolide-6-O-glucoside (10), desmethylagrimonolide (11), desmethylagrimonolide-6-O-glucoside (12), luteolin (13), vitexin (14) and isovitexin (15). Flavonoids, compound 2, 3, 11, and 14-15 have been found in APL for the first time. Furthermore, two novel flavone derivatives, compound 4 and 8, have been isolated inceptively in plant. In the no cytotoxicity concentration ranges of $0-20{\mu}M$, nitric oxide (NO) production level of 1-15 was estimated in LPS-treated Raw 264.7 macrophage cells. The flavone aglycones, 7 (apigenin, $IC_{50}=3.69{\pm}0.34{\mu}M$), 13 (luteolin, $IC_{50}=4.62{\pm}0.43{\mu}M$), 6 (kaempferol, $IC_{50}=14.43{\pm}0.23{\mu}M$) and 5 (quercetin, $IC_{50}=19.50{\pm}1.71{\mu}M$), exhibited excellent NO inhibitory (NOI) activity in dose-dependent manner. In the structure activity relationship (SAR) study of apigenin-derivatives (APD), apigenin; Api, apigenin-7-O-glucoside; Api-G, apignenin-7-O-[6"-(butyl)-glycoside]; Api-BG and apignenin-7-O-[6"-(pentyl)-glycoside]; Api-P, from APL on INF activity was investigated. The INF mediators level such as NO, INF-cytokines, NF-KB proteins, iNOS and COX-2 were sharply increased in Raw 264.7 cells by LPS. When pretreatment with APD in INF induced macrophages, NOI activity of Api was most effective than other APD with $IC_{50}$ values of $3.69{\pm}0.77{\mu}M$. And the NOI activity was declined in the following order: Api-BG ($IC_{50}=8.91{\pm}1.18{\mu}M$), Api-PG ($IC_{50}=13.52{\pm}0.85{\mu}M$) and API-G ($IC_{50}=17.30{\pm}0.66{\mu}M$). The NOI activity of two novel compounds, Api-PG and Api-BG were lower than their aglycone; Api, but more effective than Api-G (NOI: Api-PG and Api-BG). And their suppression ability on INF cytokines such as $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 mRNA showed the similar tendency. Therefore, the anti-INF mechanism study of Api-PG and Api-BG on nuclear factor-kappa B ($NF-{\kappa}B$) pathway, representative INF mechanism, was investigated and Api was used as positive control. Api-BF was more effectively prevent the than phosphorylation of $pI{\kappa}B$ kinase (p-IKK) and p65 than Api-PG in Raw 264.7 cells. In contrast, Api-PG and Api-BG were not reduced the phosphorylation of inhibitor of kappa B alpha ($I{\kappa}B{\alpha}$). Moreover, pretreatment with Api-PG and Api-BG, dose-dependently inhibited LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNAs and proteins in macrophage cells, and their expression were correlated with their NOI activity. Therefore, APL can be utilized to health promote agent associated with their AIN metabolites.

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