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1

Bibikova, Marina, and Jian‐Bing Fan. "Genome‐wide DNA methylation profiling." Wiley Interdisciplinary Reviews: Systems Biology and Medicine 2, no. 2 (2010): 210–23. http://dx.doi.org/10.1002/wsbm.35.

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Rukova, B., R. Staneva, S. Hadjidekova, G. Stamenov, V. Milanova, and D. Toncheva. "Genome-Wide Methylation Profiling of Schizophrenia." Balkan Journal of Medical Genetics 17, no. 2 (2014): 15–23. http://dx.doi.org/10.2478/bjmg-2014-0070.

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Abstract Schizophrenia is one of the major psychiatric disorders. It is a disorder of complex inheritance, involving both heritable and environmental factors. DNA methylation is an inheritable epigenetic modification that stably alters gene expression. We reasoned that genetic modifications that are a result of environmental stimuli could also make a contribution. We have performed 26 high-resolution genomewide methylation array analyses to determine the methylation status of 27,627 CpG islands and compared the data between patients and healthy controls. Methylation profiles of DNAs were analy
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Saied, Marwa, Sabah Khaled, Thomas Down, et al. "Genome Wide Study of DNA Methylation In AML." Blood 116, no. 21 (2010): 3618. http://dx.doi.org/10.1182/blood.v116.21.3618.3618.

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Abstract Abstract 3618 DNA methylation is the most stable epigenetic modification and has a major role in cancer initiation and progression. The two main aims for this research were, firstly, to use the genome wide analysis of DNA methylation to better understand the development of acute myeloid leukemia (AML). The second aim was to detect differentially methylated genes/regions between certain subtypes of AML and normal bone marrow (NBM). We used the methylated DNA immunoprecipitation technique followed by high-throughput sequencing by Illumina Genome Analyser II (MeDIP -seq) for 9 AML sample
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4

Naqvi, Hanyia, Ysabel Ilagan, Graciela Krikun, and Hugh S. Taylor. "Altered Genome-Wide Methylation in Endometriosis." Reproductive Sciences 21, no. 10 (2014): 1237–43. http://dx.doi.org/10.1177/1933719114532841.

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5

Flintoft, Louisa. "Genome-wide views of methylation readers." Nature Reviews Genetics 14, no. 6 (2013): 369. http://dx.doi.org/10.1038/nrg3509.

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6

Samarakoon, PS. "Epigenomics and Genome Wide Methylation Profiling." Sri Lanka Journal of Bio-Medical Informatics 1, no. 1 (2010): 53. http://dx.doi.org/10.4038/sljbmi.v1i1.1486.

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7

Figueroa, Maria Eugenia, John Greally, Ruud Delwel, and Ari M. Melnick. "Genome-Wide Epigenetics in Myeloid Leukemias." Blood 112, no. 11 (2008): sci—35—sci—35. http://dx.doi.org/10.1182/blood.v112.11.sci-35.sci-35.

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Abstract While the role of genetic alterations in cancer is well-recognized, epigenetic deregulation has only recently been identified as a hallmark of malignant transformation. The term “epigenetic” refers to a heritable regulation of gene expression that is not dependent on changes in the DNA sequence. These epigenetic modifications – including but not limited to DNA methylation and covalent modifications of histone tails – play a crucial role in determining chromatin structure and gene expression. Abnormal epigenetic regulation can lead to aberrant chromatin structure and deregulation of tr
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8

Lu, Tzu-Pin, Nai-Chen Chuang, Chin-Yu Cheng, et al. "Genome-wide methylation profiles in coronary artery ectasia." Clinical Science 131, no. 7 (2017): 583–94. http://dx.doi.org/10.1042/cs20160821.

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Coronary artery ectasia (CAE) is a disease characterized by abnormally dilated coronary arteries. The mechanism of CAE remains unclear, and its treatment is limited. Previous studies have shown that risk factors for CAE were related to changes in DNA methylation. However, no systematic investigation of methylation profiles has been performed. Therefore, we compared methylation profiles between 12 CAE patients and 12 propensity-matched individuals with normal coronary arteries using microarrays. Wilcoxon's rank sum tests revealed 89 genes with significantly different methylation levels (P<0.
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9

Yu, Ming, Sean K. Maden, Matthew Stachler, et al. "Subtypes of Barrett’s oesophagus and oesophageal adenocarcinoma based on genome-wide methylation analysis." Gut 68, no. 3 (2018): 389–99. http://dx.doi.org/10.1136/gutjnl-2017-314544.

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ObjectiveTo identify and characterise DNA methylation subtypes in oesophageal adenocarcinoma (EAC) and its precursor Barrett’s oesophagus (BE).DesignWe performed genome-wide DNA methylation profiling on samples of non-dysplastic BE from cancer-free patients (n=59), EAC (n=23), normal squamous oesophagus (n=33) and normal fundus (n=9), and identified methylation subtypes using a recursively partitioned mixture model. We assessed genomic alterations for 9 BE and 22 EAC samples with massively parallel sequencing of 243 EAC-associated genes, and we conducted integrative analyses with transcriptome
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10

Chandrasekhar, Manikala, Anilkumar Chandrappa, Rameswar Prasad Sah, et al. "AMP-PCR-based assay for detection and quantification of genome wide natural methylation in rice." Indian Journal of Genetics and Plant Breeding (The) 84, no. 04 (2024): 635–43. https://doi.org/10.31742/isgpb.84.4.14.

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Natural and artificial selection efforts combined several favorable alleles of economically important traits in crop plants. However, the progress made is insufficient to meet the future food requirements. Hence, exploring new genetic resources and breeding strategies is important for sustainable improvement in production. The epigenetic variation that alters the phenotype expression without altering the gene sequence has played a crucial role in the process of evolution of modern-day crop plants. The methylation-based epigenetic variations are known to inherit more consistently than other typ
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11

Osumi, Hiroki, Kota Ouchi, Eiji Shinozaki, et al. "Clinical impact of DNA methylation status on first-line antiepidermal growth factor receptor treatment in patients with metastatic colorectal cancer." Journal of Clinical Oncology 40, no. 16_suppl (2022): 3528. http://dx.doi.org/10.1200/jco.2022.40.16_suppl.3528.

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3528 Background: The CpG island methylator phenotype (CIMP), important for carcinogenesis, is a predictor of prognosis and chemotherapy sensitivity in colorectal cancer. However, there is a lack of consensus of CIMP markers, and thus, more comprehensive methylation markers are required to reliably predict the clinical outcomes. Previously, we reported that genome-wide DNAmethylation statuscould predict the effect of epidermal growth factor receptor (EGFR) inhibitors more accurately than previously reported methylation classifications (Ouchi et al. Cancer Sci 2015). Moreover, we had developed a
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12

Michaelson-Cohen, Rachel, Ilana Keshet, Ravid Straussman, Merav Hecht, Howard Cedar, and Uziel Beller. "Genome-Wide De Novo Methylation in Epithelial Ovarian Cancer." International Journal of Gynecologic Cancer 21, no. 2 (2011): 269–79. http://dx.doi.org/10.1097/igc.0b013e31820e5cda.

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Background:DNA methylation regulates gene expression during development. The methylation pattern is established at the time of implantation. CpG islands are genome regions usually protected from methylation; however, selected islands are methylated later. Many undergo methylation in cancer, causing epigenetic gene silencing. Aberrant methylation occurs early in tumorigenesis, in a specific pattern, inhibiting differentiation.Although methylation of specific genes in ovarian tumors has been demonstrated in numerous studies, they represent only a fraction of all methylated genes in tumorigenesis
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13

Wei, Zhen, Subbarayalu Panneerdoss, Santosh Timilsina, et al. "Topological Characterization of Human and Mouse m5C Epitranscriptome Revealed by Bisulfite Sequencing." International Journal of Genomics 2018 (June 13, 2018): 1–19. http://dx.doi.org/10.1155/2018/1351964.

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Background. Compared with the well-studied 5-methylcytosine (m5C) in DNA, the role and topology of epitranscriptome m5C remain insufficiently characterized. Results. Through analyzing transcriptome-wide m5C distribution in human and mouse, we show that the m5C modification is significantly enriched at 5′ untranslated regions (5′UTRs) of mRNA in human and mouse. With a comparative analysis of the mRNA and DNA methylome, we demonstrate that, like DNA methylation, transcriptome m5C methylation exhibits a strong clustering effect. Surprisingly, an inverse correlation between mRNA and DNA m5C methy
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14

Lewsey, Mathew G., Thomas J. Hardcastle, Charles W. Melnyk, et al. "Mobile small RNAs regulate genome-wide DNA methylation." Proceedings of the National Academy of Sciences 113, no. 6 (2016): E801—E810. http://dx.doi.org/10.1073/pnas.1515072113.

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RNA silencing at the transcriptional and posttranscriptional levels regulates endogenous gene expression, controls invading transposable elements (TEs), and protects the cell against viruses. Key components of the mechanism are small RNAs (sRNAs) of 21–24 nt that guide the silencing machinery to their nucleic acid targets in a nucleotide sequence-specific manner. Transcriptional gene silencing is associated with 24-nt sRNAs and RNA-directed DNA methylation (RdDM) at cytosine residues in three DNA sequence contexts (CG, CHG, and CHH). We previously demonstrated that 24-nt sRNAs are mobile from
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15

Arribas, Alberto J., Andrea Rinaldi, Giorgia Chiodin, et al. "Genome-wide promoter methylation of hairy cell leukemia." Blood Advances 3, no. 3 (2019): 384–96. http://dx.doi.org/10.1182/bloodadvances.2018024059.

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Abstract Classic hairy cell leukemia (HCL) is a tumor of mature clonal B cells with unique genetic, morphologic, and phenotypic features. DNA methylation profiling has provided a new tier of investigation to gain insight into the origin and behavior of B-cell malignancies; however, the methylation profile of HCL has not been specifically investigated. DNA methylation profiling was analyzed with the Infinium HumanMethylation27 array in 41 mature B-cell tumors, including 11 HCL, 7 splenic marginal zone lymphomas (SMZLs), and chronic lymphocytic leukemia with an unmutated (n = 7) or mutated (n =
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16

Devuyst, Olivier. "Genome-Wide Methylation and Body-Mass Index." Peritoneal Dialysis International: Journal of the International Society for Peritoneal Dialysis 34, no. 5 (2014): 477. http://dx.doi.org/10.3747/pdi.2014.00181.

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17

Bhatti, Parveen, Yuzheng Zhang, Xiaoling Song, et al. "Nightshift work and genome-wide DNA methylation." Chronobiology International 32, no. 1 (2014): 103–12. http://dx.doi.org/10.3109/07420528.2014.956362.

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18

Xu, Xiaojing, Shaoyong Su, Vernon A. Barnes, et al. "A genome-wide methylation study on obesity." Epigenetics 8, no. 5 (2013): 522–33. http://dx.doi.org/10.4161/epi.24506.

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19

Gupta*, Romi, Arvindhan Nagarajan*, and Narendra Wajapeyee. "Advances in genome-wide DNA methylation analysis." BioTechniques 49, no. 4 (2010): iii—xi. http://dx.doi.org/10.2144/000113493.

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20

Zilberman, D., and S. Henikoff. "Genome-wide analysis of DNA methylation patterns." Development 134, no. 22 (2007): 3959–65. http://dx.doi.org/10.1242/dev.001131.

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21

Lai, Rose K., Yanwen Chen, Xiaowei Guan, et al. "Genome-Wide Methylation Analyses in Glioblastoma Multiforme." PLoS ONE 9, no. 2 (2014): e89376. http://dx.doi.org/10.1371/journal.pone.0089376.

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22

FERNÁNDEZ-SANLÉS, ALBA, SERGI SAYOLS-BAIXERAS, MANUEL CASTRO DE MOURA, et al. "Physical Activity and Genome-wide DNA Methylation." Medicine & Science in Sports & Exercise 52, no. 3 (2020): 589–97. http://dx.doi.org/10.1249/mss.0000000000002174.

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23

Zhang, Michael Q., and Andrew D. Smith. "Challenges in Understanding Genome-Wide DNA Methylation." Journal of Computer Science and Technology 25, no. 1 (2010): 26–34. http://dx.doi.org/10.1007/s11390-010-9302-8.

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24

Montano, Carolina, and Winston Timp. "Evolution of genome-wide methylation profiling technologies." Genome Research 35, no. 4 (2025): 572–82. https://doi.org/10.1101/gr.278407.123.

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In this mini-review, we explore the advancements in genome-wide DNA methylation profiling, tracing the evolution from traditional methods such as methylation arrays and whole-genome bisulfite sequencing to the cutting-edge single-molecule profiling enabled by long-read sequencing (LRS) technologies. We highlight how LRS is transforming clinical and translational research, particularly by its ability to simultaneously measure genetic and epigenetic information, providing a more comprehensive understanding of complex disease mechanisms. We discuss current challenges and future directions in the
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25

Ili, Carmen, Kurt Buchegger, Hannah Demond, et al. "Landscape of Genome-Wide DNA Methylation of Colorectal Cancer Metastasis." Cancers 12, no. 9 (2020): 2710. http://dx.doi.org/10.3390/cancers12092710.

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Colorectal cancer is a heterogeneous disease caused by both genetic and epigenetics factors. Analysing DNA methylation changes occurring during colorectal cancer progression and metastasis formation is crucial for the identification of novel epigenetic markers of patient prognosis. Genome-wide methylation sequencing of paired samples of colon (normal adjacent, primary tumour and lymph node metastasis) showed global hypomethylation and CpG island (CGI) hypermethylation of primary tumours compared to normal. In metastasis we observed high global and non-CGI regions methylation, but lower CGI met
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26

Chen, Xi, Liu-Hong Shen, Li-Xuan Gui, et al. "Genome-wide DNA methylation profile of prepubertal porcine testis." Reproduction, Fertility and Development 30, no. 2 (2018): 349. http://dx.doi.org/10.1071/rd17067.

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The biological structure and function of the mammalian testis undergo important developmental changes during prepuberty and DNA methylation is dynamically regulated during testis development. In this study, we generated the first genome-wide DNA methylation profile of prepubertal porcine testis using methyl-DNA immunoprecipitation (MeDIP) combined with high-throughput sequencing (MeDIP-seq). Over 190 million high-quality reads were generated, containing 43 642 CpG islands. There was an overall downtrend of methylation during development, which was clear in promoter regions but less so in gene-
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27

Kim, Jinsil, Mitchell M. Pitlick, Paul J. Christine, et al. "Genome-Wide Analysis of DNA Methylation in Human Amnion." Scientific World Journal 2013 (2013): 1–11. http://dx.doi.org/10.1155/2013/678156.

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The amnion is a specialized tissue in contact with the amniotic fluid, which is in a constantly changing state. To investigate the importance of epigenetic events in this tissue in the physiology and pathophysiology of pregnancy, we performed genome-wide DNA methylation profiling of human amnion from term (with and without labor) and preterm deliveries. Using the Illumina Infinium HumanMethylation27 BeadChip, we identified genes exhibiting differential methylation associated with normal labor and preterm birth. Functional analysis of the differentially methylated genes revealed biologically re
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Jevšinek Skok, Daša, and Nina Hauptman. "In Silico Gene Prioritization Highlights the Significance of Bone Morphogenetic Protein 4 (BMP4) Promoter Methylation across All Methylation Clusters in Colorectal Cancer." International Journal of Molecular Sciences 24, no. 16 (2023): 12692. http://dx.doi.org/10.3390/ijms241612692.

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The cytosine–phosphate–guanine (CpG) island methylator phenotype (CIMP) represents one of the pathways involved in the development of colorectal cancer, characterized by genome-wide hypermethylation. To identify samples exhibiting hypermethylation, we used unsupervised hierarchical clustering on genome-wide methylation data. This clustering analysis revealed the presence of four distinct subtypes within the tumor samples, namely, CIMP-H, CIMP-L, cluster 3, and cluster 4. These subtypes demonstrated varying levels of methylation, categorized as high, intermediate, and very low. To gain further
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van der Plaat, Diana A., Kim de Jong, Maaike de Vries, et al. "Occupational exposure to pesticides is associated with differential DNA methylation." Occupational and Environmental Medicine 75, no. 6 (2018): 427–35. http://dx.doi.org/10.1136/oemed-2017-104787.

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ObjectivesOccupational pesticide exposure is associated with a wide range of diseases, including lung diseases, but it is largely unknown how pesticides influence airway disease pathogenesis. A potential mechanism might be through epigenetic mechanisms, like DNA methylation. Therefore, we assessed associations between occupational exposure to pesticides and genome-wide DNA methylation sites.Methods1561 subjects of LifeLines were included with either no (n=1392), low (n=108) or high (n=61) exposure to any type of pesticides (estimated based on current or last held job). Blood DNA methylation le
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30

Decock, Anneleen, Maté Ongenaert, Jasmien Hoebeeck, et al. "Genome-wide promoter methylation analysis in neuroblastoma identifies prognostic methylation biomarkers." Genome Biology 13, no. 10 (2012): R95. http://dx.doi.org/10.1186/gb-2012-13-10-r95.

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31

Sujit, Kumar Mohanty, Saumya Sarkar, Vertika Singh, et al. "Genome-wide differential methylation analyses identifies methylation signatures of male infertility." Human Reproduction 33, no. 12 (2018): 2256–67. http://dx.doi.org/10.1093/humrep/dey319.

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32

Yao, Haiqiang, Shanlan Mo, Ji Wang, et al. "Genome-Wide DNA Methylation Profiles of Phlegm-Dampness Constitution." Cellular Physiology and Biochemistry 45, no. 5 (2018): 1999–2008. http://dx.doi.org/10.1159/000487976.

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Background/Aims: Metabolic diseases are leading health concerns in today’s global society. In traditional Chinese medicine (TCM), one body type studied is the phlegm-dampness constitution (PC), which predisposes individuals to complex metabolic disorders. Genomic studies have revealed the potential metabolic disorders and the molecular features of PC. The role of epigenetics in the regulation of PC, however, is unknown. Methods: We analyzed a genome-wide DNA methylation in 12 volunteers using Illumina Infinium Human Methylation450 BeadChip on peripheral blood mononuclear cells (PBMCs). Eight v
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Brenet, Fabienne, Michelle Moh, Patricia Funk, et al. "Genome-Wide Analysis of DNA Methylation Patterns Reveals Dynamic Epigenetic Regulation of the AML Genome After Decitabine Treatment." Blood 114, no. 22 (2009): 591. http://dx.doi.org/10.1182/blood.v114.22.591.591.

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Abstract Abstract 591 The human genome is adorned with methylated cytosine residues that function in the epigenetic guidance of cellular differentiation and development. Cellular interpretation of this epigenetic mark is incompletely understood and tissue specific patterns of DNA methylation vary with age, can be altered by environmental factors, and are often abnormal in human disease. Aberrant DNA methylation is a common means by which tumor suppressor genes (TSGs) are inactivated during carcinogenesis (Baylin, Herman, Graff, Vertino and Issa 1998; Laird and Jaenisch 1996; Singal and Ginder
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34

Zhong, Sheng L., Yuanyuan Xiao, Ru-Fang Yeh, et al. "Genome-Wide Scanning of Epigenomic Alterations in Childhood Leukemia." Blood 112, no. 11 (2008): 4464. http://dx.doi.org/10.1182/blood.v112.11.4464.4464.

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Abstract To study how epigenetic changes contribute to childhood leukemia, and to identify new disease biomarkers, we combined an approach of pharmacologic inhibition of epigenetic modifications of chromatin with gene expression arrays (Affymetrix Exon 1.0; and miRNA array, UCSF-designed) to reveal precise epigenetic aberrations on a global scale in defined subsets of leukemia cell lines that recapitulate common genetic subtypes of leukemia. From these analyses, we identified >200 genes up-regulated by 5-aza-2′-deoxycytidine and trichostatin A treatment (including genes encoding both mR
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35

Movassagh, Mehregan, Ana Vujic, and Roger Foo. "Genome-wide DNA methylation in human heart failure." Epigenomics 3, no. 1 (2011): 103–9. http://dx.doi.org/10.2217/epi.10.70.

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36

YAMADA, NOBUHISA, KOHICHIROH YASUI, OSAMU DOHI, et al. "Genome-wide DNA methylation analysis in hepatocellular carcinoma." Oncology Reports 35, no. 4 (2016): 2228–36. http://dx.doi.org/10.3892/or.2016.4619.

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37

Ferfouri, F., F. Boitrelle, I. Ghout, et al. "A genome-wide DNA methylation study in azoospermia." Andrology 1, no. 6 (2013): 815–21. http://dx.doi.org/10.1111/j.2047-2927.2013.00117.x.

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38

Zemach, A., I. E. McDaniel, P. Silva, and D. Zilberman. "Genome-Wide Evolutionary Analysis of Eukaryotic DNA Methylation." Science 328, no. 5980 (2010): 916–19. http://dx.doi.org/10.1126/science.1186366.

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39

Kaneda, Ruri, Shuji Takada, Yoshihiro Yamashita, et al. "Genome-wide histone methylation profile for heart failure." Genes to Cells 14, no. 1 (2009): 69–77. http://dx.doi.org/10.1111/j.1365-2443.2008.01252.x.

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40

Fleischer, Thomas, Arnoldo Frigessi, Kevin C. Johnson, et al. "Genome-wide DNA methylation profiles in progression to." Genome Biology 15, no. 8 (2014): 435. http://dx.doi.org/10.1186/preaccept-2333349012841587.

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41

Yau, M. S., R. Joehanes, Y. H. Hsu, D. P. Kiel, and D. T. Felson. "Genome-wide DNA methylation study of hand osteoarthritis." Osteoarthritis and Cartilage 26 (April 2018): S158. http://dx.doi.org/10.1016/j.joca.2018.02.342.

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42

Li, Qinghe, Ning Li, Xiaoxiang Hu, et al. "Genome-Wide Mapping of DNA Methylation in Chicken." PLoS ONE 6, no. 5 (2011): e19428. http://dx.doi.org/10.1371/journal.pone.0019428.

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43

Cai, Tian-tian, Fatuma-said Muhali, Rong-hua Song, et al. "Genome-wide DNA methylation analysis in Graves' disease." Genomics 105, no. 4 (2015): 204–10. http://dx.doi.org/10.1016/j.ygeno.2015.01.001.

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Song, Qing-Xin, Xiang Lu, Qing-Tian Li, et al. "Genome-Wide Analysis of DNA Methylation in Soybean." Molecular Plant 6, no. 6 (2013): 1961–74. http://dx.doi.org/10.1093/mp/sst123.

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Jones, Lamont R., William Young, George Divine, et al. "Genome-Wide Scan for Methylation Profiles in Keloids." Disease Markers 2015 (2015): 1–7. http://dx.doi.org/10.1155/2015/943176.

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Keloids are benign fibroproliferative tumors of the skin which commonly occur after injury mainly in darker skinned patients. Medical treatment is fraught with high recurrence rates mainly because of an incomplete understanding of the biological mechanisms that lead to keloids. The purpose of this project was to examine keloid pathogenesis from the epigenome perspective of DNA methylation. Genome-wide profiling used the Infinium HumanMethylation450 BeadChip to interrogate DNA from 6 fresh keloid and 6 normal skin samples from 12 anonymous donors. A 3-tiered approach was used to call out genes
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46

Zhang, Ruiling, Qin Miao, Chuansheng Wang, et al. "Genome-wide DNA methylation analysis in alcohol dependence." Addiction Biology 18, no. 2 (2013): 392–403. http://dx.doi.org/10.1111/adb.12037.

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47

Hatada, I., M. Fukasawa, M. Kimura, et al. "Genome-wide profiling of promoter methylation in human." Oncogene 25, no. 21 (2006): 3059–64. http://dx.doi.org/10.1038/sj.onc.1209331.

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48

Ananiev, Gene E., Steve Goldstein, Rod Runnheim, et al. "Optical mapping discerns genome wide DNA methylation profiles." BMC Molecular Biology 9, no. 1 (2008): 68. http://dx.doi.org/10.1186/1471-2199-9-68.

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Shen, Jing, Shuang Wang, Yu-Jing Zhang, et al. "Genome-wide DNA methylation profiles in hepatocellular carcinoma." Hepatology 55, no. 6 (2012): 1799–808. http://dx.doi.org/10.1002/hep.25569.

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50

Dai, Chao, Giancarlo Bonora, Kemin Zhou, Shidong Jia, and Pan Du. "Abstract 4557: Genome-wide cfDNA hypo-methylation landscape of 500 patients, a real-world study." Cancer Research 85, no. 8_Supplement_1 (2025): 4557. https://doi.org/10.1158/1538-7445.am2025-4557.

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Abstract Background: Liquid biopsies analyzing cell-free DNA (cfDNA) from plasma and urine can offer clinical diagnostic information in a non-invasive way. One major application of DNA methylation in liquid biopsies focuses on targeted panel regions, aiming to detect hypermethylation in cancer related regions. Recent studies have revealed that the loss of DNA methylation is significantly enriched in late-replicating loci compared with early-replicating loci in cancer cells. In this study, we profiled genome-wide DNA methylation of plasma and urine samples from various of cancer indications to
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