• Title/Summary/Keyword: RNA sequencing

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Whole genome sequence of Staphylococcus aureus strain RMI-014804 isolated from pulmonary patient sputum via next-generation sequencing technology

  • Ayesha, Wisal;Asad Ullah;Waheed Anwar;Carlos M. Morel;Syed Shah Hassan
    • Genomics & Informatics
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    • v.21 no.3
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    • pp.34.1-34.10
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    • 2023
  • Nosocomial infections, commonly referred to as healthcare-associated infections, are illnesses that patients get while hospitalized and are typically either not yet manifest or may develop. One of the most prevalent nosocomial diseases in hospitalized patients is pneumonia, among the leading causes of mortality and morbidity. Viral, bacterial, and fungal pathogens cause pneumonia. More severe introductions commonly included Staphylococcus aureus, which is at the top of bacterial infections, per World Health Organization reports. The staphylococci, S. aureus, strain RMI-014804, mesophile, on-sporulating, and non-motile bacterium, was isolated from the sputum of a pulmonary patient in Pakistan. Many characteristics of S. aureus strain RMI-014804 have been revealed in this paper, with complete genome sequence and annotation. Our findings indicate that the genome is a single circular 2.82 Mbp long genome with 1,962 protein-coding genes, 15 rRNA, 49 tRNA, 62 pseudogenes, and a GC content of 28.76%. As a result of this genome sequencing analysis, researchers will fully understand the genetic and molecular basis of the virulence of the S. aureus bacteria, which could help prevent the spread of nosocomial infections like pneumonia. Genome analysis of this strain was necessary to identify the specific genes and molecular mechanisms that contribute to its pathogenicity, antibiotic resistance, and genetic diversity, allowing for a more in-depth investigation of its pathogenesis to develop new treatments and preventive measures against infections caused by this bacterium.

Hepatic microRNAome reveals potential microRNA-mRNA pairs association with lipid metabolism in pigs

  • Liu, Jingge;Ning, Caibo;Li, Bojiang;Li, Rongyang;Wu, Wangjun;Liu, Honglin
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.9
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    • pp.1458-1468
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    • 2019
  • Objective: As one of the most important metabolic organs, the liver plays vital roles in modulating the lipid metabolism. This study was to compare miRNA expression profiles of the Large White liver between two different developmental periods and to identify candidate miRNAs for lipid metabolism. Methods: Eight liver samples were collected from White Large of 70-day fetus (P70) and of 70-day piglets (D70) (with 4 biological repeats at each development period) to construct sRNA libraries. Then the eight prepared sRNA libraries were sequenced using Illumina next-generation sequencing technology on HiSeq 2500 platform. Results: As a result, we obtained 346 known and 187 novel miRNAs. Compared with the D70, 55 down- and 61 up-regulated miRNAs were shown to be significantly differentially expressed (DE). Gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis indicated that these DE miRNAs were mainly involved in growth, development and diverse metabolic processes. They were predicted to regulate lipid metabolism through adipocytokine signaling pathway, mitogen-activated protein kinase, AMP-activated protein kinase, cyclic adenosine monophosphate, phosphatidylinositol 3 kinase/protein kinase B, and Notch signaling pathway. The four most abundantly expressed miRNAs were miR-122, miR-26a and miR-30a-5p (miR-122 only in P70), which play important roles in lipid metabolism. Integration analysis (details of mRNAs sequencing data were shown in another unpublished paper) revealed that many target genes of the DE miRNAs (miR-181b, miR-145-5p, miR-199a-5p, and miR-98) might be critical regulators in lipid metabolic process, including acyl-CoA synthetase long chain family member 4, ATP-binding casette A4, and stearyl-CoA desaturase. Thus, these miRNAs were the promising candidates for lipid metabolism. Conclusion: Our study provides the main differences in the Large White at miRNA level between two different developmental stages. It supplies a valuable database for the further function and mechanism elucidation of miRNAs in porcine liver development and lipid metabolism.

Variable Selection of Feature Pattern using SVM-based Criterion with Q-Learning in Reinforcement Learning (SVM-기반 제약 조건과 강화학습의 Q-learning을 이용한 변별력이 확실한 특징 패턴 선택)

  • Kim, Chayoung
    • Journal of Internet Computing and Services
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    • v.20 no.4
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    • pp.21-27
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    • 2019
  • Selection of feature pattern gathered from the observation of the RNA sequencing data (RNA-seq) are not all equally informative for identification of differential expressions: some of them may be noisy, correlated or irrelevant because of redundancy in Big-Data sets. Variable selection of feature pattern aims at differential expressed gene set that is significantly relevant for a special task. This issues are complex and important in many domains, for example. In terms of a computational research field of machine learning, selection of feature pattern has been studied such as Random Forest, K-Nearest and Support Vector Machine (SVM). One of most the well-known machine learning algorithms is SVM, which is classical as well as original. The one of a member of SVM-criterion is Support Vector Machine-Recursive Feature Elimination (SVM-RFE), which have been utilized in our research work. We propose a novel algorithm of the SVM-RFE with Q-learning in reinforcement learning for better variable selection of feature pattern. By comparing our proposed algorithm with the well-known SVM-RFE combining Welch' T in published data, our result can show that the criterion from weight vector of SVM-RFE enhanced by Q-learning has been improved by an off-policy by a more exploratory scheme of Q-learning.

hnRNPK-regulated PTOV1-AS1 modulates heme oxygenase-1 expression via miR-1207-5p

  • Shin, Chang Hoon;Ryu, Seongho;Kim, Hyeon Ho
    • BMB Reports
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    • v.50 no.4
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    • pp.220-225
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    • 2017
  • Antisense transcripts were initially identified as transcriptional noise, but have since been reported to play an important role in the quality control of miRNA functions. In this report, we tested the hypothesis that heterogeneous nuclear ribonucleoprotein K (hnRNPK) regulates miRNA function via competitive endogenous RNAs, such as pseudogenes, long non-coding RNAs, and antisense transcripts. Based on analyses of RNA sequencing data, the knockdown of hnRNPK decreased the antisense PTOV1-AS1 transcript which harbors five binding sites for miR-1207-5p. We identified heme oxygenase-1 (HO-1) mRNA as a novel target of miR-1207-5p by western blotting and Ago2 immunoprecipitation. The knockdown of hnRNPK or PTOV1-AS1 suppressed HO-1 expression by increasing the enrichment of HO-1 mRNA in miR-1207-5p-mediated miRISC. Downregulation of HO-1 by a miR-1207-5p mimic or knockdown of hnRNPK and PTOV1-AS1 inhibited the proliferation and clonogenic ability of HeLa cells. Taken together, our results demonstrate that hnRNPK-regulated PTOV1-AS1 modulates HO-1 expression via miR-1207-5p.

Isolation and Identification of Genes Regulated by Iron Using mRNA Differential Display (mRNA differential display를 이용한 철에 의해 조절되는 유전자들의 분리 및 동정)

  • Lee, Jung-Lim;Park, Jong-Hwan;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.42 no.2
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    • pp.123-127
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    • 1999
  • Iron is an essential nutrient but potentially toxic element in human. To identify the effects of iron on the gene expression of mammalian cell, we have isolated several genes that are regulated by iron using the RNA differential display method. RNAs were isolated from HeLa cells treated with iron supplement or iron chelator. A total of 24 genes were isolated and of these, four genes were identified by DNA sequencing and northern blot.

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Human Caliciviruses in Korea: A New Prevalent Group Defined by RNA-Dependent RNA Polymerase Diversity (한국형 사람 Caliciviruses의 RNA-Dependent RNA Polymerase Diversity)

  • Han, Dong-Pyo;Kim, Ji-Aee;Yang, Jai-Myung;Kim, Kyung-Hee
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.1-8
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    • 1997
  • Human caliciviruses (HuCVs) cause sporadic cases and outbreaks of acute gastroenteritis (AGE). Three major genogroups of HuCVs have been described including the Norwalk virus (NV)-, the Snow Mountain virus (SMA)-, and the Sapporo-genogroups. This study describes the detection and genetic variation of HuCVs from hospitalized infants with AGE in Korea by RT-PCR and sequencing. The cDNA fragments of 206 to 470bp corresponding to the region of 3 primer pairs (36/35, 35/51 or 3/51) in the polymerase region of NV were generated. Of 185 stools screened, 8% were positive by RT-PCR and their sequences showed that all strains contained the GLPSG and YGDD motifs which are conserved for HuCVs. Amino acid (aa) sequence analysis showed that these strains can be divided into 3 major genogroups. High conservation was observed in that one strain shares 100% of aa sequence with Southampton virus, another shares 99% with the Sapporo virus, and six strains share 90 to 95% with Snow Mountain virus. However, significant sequence variation was also found in other strains. This study indicates that all major genogroups of HuCVs are circulating in Korea.

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Genetic characterization and phylogenetic analysis of Clostridium chauvoei isolated from Hanwoo in Jeonbuk (전북지역 한우에서 분리한 기종저 균의 유전학적 특성 규명)

  • Kim, Chul-Min;Jeong, Jae-Myong;Choi, Ki-Young
    • Korean Journal of Veterinary Service
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    • v.37 no.3
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    • pp.157-164
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    • 2014
  • Clostridium chauvoei is the etiologic agent of blackleg, a high mortality rated disease infection mainly cattle. In the present study, the partial sequences of 16S rRNA and flagellin gene of C. chauvoei isolated in Jeonbuk, Korea were determined and compared with those of reference strain. Oligonucleotide primers were designed to amplify a 811 bp fragment of 16S rRNA gene and 1229 bp fragment of flagellin gene. Sequencing analysis of 16S rRNA gene showed high homology to the reference strains ranging 82.3% to 100%, while flagellin gene were different from published foreign clostridia, showing 98.7% to 72.0% nucleotide sequence homology. Phylogenetic analysis based on 16S rRNA gene revealed the close phylogenetic relationship of C. chauvoei and C. septicum in cluster I, which includes C. carnis, C. tertium, C. quinii, C. celatum, C. perfringens, C. absonum, C. botulinum B. Phylogentic analysis also revealed that flagellin gene formed a single cluster with C. chauvoei, C. septicum, C. novyi A, C. novyi B, C. tyrobutylicum, C. acetobutylicum. The genetic informations obtained from this study could be useful for the molecular study of C. chauvoei.

Evolutionary Relationships of the Genus Trichoderma and Related Taxa Based on the Partial Sequences of 18S Ribosomal RNA (18S 리보좀 RNA 부분 염기서열에 의한 Trichoderma속 및 관련 불완전균류의 진화학적 유연관계)

  • Lee, Goang-Jae;An, Won-Gun;Lee, Jae-Dong;Joo, Woo-Hong
    • The Korean Journal of Mycology
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    • v.23 no.4 s.75
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    • pp.318-324
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    • 1995
  • The evolutionary relationships of the genus Trichoderma and related taxa were assessed using partial sequencing of 18S ribosomal RNA. Phylogenetic tree divided into three major groups; 1. Saccharomyces cerevisiae-Geotrichum klebahnii-Alternaria mali group; 2. Neurospora crassa-Aspergillus-Penicillium-Chrysosporium pannorum-Scopulariopsis sp. group; 3. Trichoderma group. The genus Trichoderma seemed to be phylogenetically separated from Saccharomyces cerevisiae, Aspergillus and Penicillium groups, and have passed through it's own evolutionary pathway.

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Plant RNA Virus Sequences Identified in Kimchi by Microbial Metatranscriptome Analysis

  • Kim, Dong Seon;Jung, Ji Young;Wang, Yao;Oh, Hye Ji;Choi, Dongjin;Jeon, Che Ok;Hahn, Yoonsoo
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.979-986
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    • 2014
  • Plant pathogenic RNA viruses are present in a variety of plant-based foods. When ingested by humans, these viruses can survive the passage through the digestive tract, and are frequently detected in human feces. Kimchi is a traditional fermented Korean food made from cabbage or vegetables, with a variety of other plant-based ingredients, including ground red pepper and garlic paste. We analyzed microbial metatranscriptome data from kimchi at five fermentation stages to identify plant RNA virus-derived sequences. We successfully identified a substantial amount of plant RNA virus sequences, especially during the early stages of fermentation: 23.47% and 16.45% of total clean reads on days 7 and 13, respectively. The most abundant plant RNA virus sequences were from pepper mild mottle virus, a major pathogen of red peppers; this constituted 95% of the total RNA virus sequences identified throughout the fermentation period. We observed distinct sequencing read-depth distributions for plant RNA virus genomes, possibly implying intrinsic and/or technical biases during the metatranscriptome generation procedure. We also identified RNA virus sequences in publicly available microbial metatranscriptome data sets. We propose that metatranscriptome data may serve as a valuable resource for RNA virus detection, and a systematic screening of the ingredients may help prevent the use of virus-infected low-quality materials for food production.

MicroRNA Regulation in Systemic Lupus Erythematosus Pathogenesis

  • Yan, Sheng;Yim, Lok Yan;Lu, Liwei;Lau, Chak Sing;Chan, Vera Sau-Fong
    • IMMUNE NETWORK
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    • v.14 no.3
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    • pp.138-148
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    • 2014
  • MicroRNAs (miRNAs) are endogenous small RNA molecules best known for their function in post-transcriptional gene regulation. Immunologically, miRNA regulates the differentiation and function of immune cells and its malfunction contributes to the development of various autoimmune diseases including systemic lupus erythematosus (SLE). Over the last decade, accumulating researches provide evidence for the connection between dysregulated miRNA network and autoimmunity. Interruption of miRNA biogenesis machinery contributes to the abnormal T and B cell development and particularly a reduced suppressive function of regulatory T cells, leading to systemic autoimmune diseases. Additionally, multiple factors under autoimmune conditions interfere with miRNA generation via key miRNA processing enzymes, thus further skewing the miRNA expression profile. Indeed, several independent miRNA profiling studies reported significant differences between SLE patients and healthy controls. Despite the lack of a consistent expression pattern on individual dysregulated miRNAs in SLE among these studies, the aberrant expression of distinct groups of miRNAs causes overlapping functional outcomes including perturbed type I interferon signalling cascade, DNA hypomethylation and hyperactivation of T and B cells. The impact of specific miRNA-mediated regulation on function of major immune cells in lupus is also discussed. Although research on the clinical application of miRNAs is still immature, through an integrated approach with advances in next generation sequencing, novel tools in bioinformatics database analysis and new in vitro and in vivo models for functional evaluation, the diagnostic and therapeutic potentials of miRNAs may bring to fruition in the future.