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Methods in molecular exercise physiology
Published in Adam P. Sharples, James P. Morton, Henning Wackerhage, Molecular Exercise Physiology, 2022
Adam P. Sharples, Daniel C. Turner, Stephen Roth, Robert A. Seaborne, Brendan Egan, Mark Viggars, Jonathan C. Jarvis, Daniel J. Owens, Jatin G. Burniston, Piotr P. Gorski, Claire E. Stewart
For a genome-wide approach, ChIP-on-chip was the first widespread method in which DNA fragments are analysed via a microarray analysis. However, with rapid development of NGS, ChIP-sequencing has become the more common methodology employed. In this process, DNA fragments are purified from the ChIP protocol detailed above and used for generating DNA sequencing libraries. Nonetheless, both approaches provide information regarding the binding location and amount of binding of the histone modification of choice (e.g. the histone mark used for the antibody immunoprecipitation step) (29). Each of these protocols however is costly, and produces a large amount of data and requires advanced expertise in bioinformatics and data analysis (30). To overcome these limitations, researchers are now able to analyse the purified DNA fragments via quantitative PCR (qPCR), in a far more targeted approach. This protocol relies heavily on the researchers knowing exactly the region in the genome they wish to target, and the ability to design PCR primers to investigate this area. ChIP protocols require large quantities of starting material, are relatively labour-intensive and, for sequencing protocols, require large amounts of sequencing data in order to reliably determine meaningful differences across datasets.
Role of Histone Methyltransferase in Breast Cancer
Published in Meenu Gupta, Rachna Jain, Arun Solanki, Fadi Al-Turjman, Cancer Prediction for Industrial IoT 4.0: A Machine Learning Perspective, 2021
Surekha Manhas, Zaved Ahmed Khan
G9a’s role also has implications in various aspects associated with T-cell-based biology. However, genome mapping studies on the G9a binding or H3K9me2 deposition in immunity-providing cells has not performed due to some technical issues. A comprehensive descriptive H3K9me2 marks genome analysis in the case of resting immune cells by means of using ChIP-on-chip based methods, which illustrate that the epigenetic H3K9me2 mark is highly enriched on those genes that are strongly associated with numerous specific pathways, such as GATA3 transcription, T-cell-dependent receptor-based signaling, and IL-4 signaling [60].
Falling Through the Safety Net
Published in Kant Patel, Mark Rushefsky, Healthcare Politics and Policy in America, 2019
According to the Pew Hispanic Center, most undocumented immigrants in the United States are employed in the agriculture sector (17 percent), followed by construction (13 percent), leisure/hospitality (7 percent), and manufacturing (6 percent) (Passel and Cohn 2017). Most undocumented immigrants, given their immigration status and employment concentration in certain sectors of the economy, earn close to minimum wage and fall below the federal poverty line. However, given their status as undocumented immigrants they are ineligible for Medicaid and the CHIP (“Medicaid/CHIP Coverage of Lawfully-Residing Immigrant Children and Pregnant Women” 2018).
Effectiveness of bone substitute materials in opening wedge high tibial osteotomy: a systematic review and meta-analysis
Published in Annals of Medicine, 2022
Tao Bei, Liping Yang, Qiulin Huang, Jiaheng Wu, Junting Liu
Among the included studies, there were five RCTs [20–24] and eight NRS including one prospective study [25], one non-inferiority study [26] and six retrospective studies [16,27–31]. The included studies were conducted between 2010 and 2021 and involved 373 patients treated using BSM, 77 patients treated with AU, 164 patients treated with AL and 155 patients treated with no graft. The average age of the patients ranged from 44 to 61 years. The fixing plates utilized were LLP, SLP, short spacer plate (SSP) and short spacer locked plate (SSLP). The average follow-up period ranged from 6 to 24 months. In a retrospective study by Kim et al. [31], 97 knees were divided into three groups, which were assigned to treat with HA chip, allogenic chip bone (AL) or WG. We separated the three groups into two comparative groups: HA vs. AL and HA vs. WG. The characteristics of the included studies are summarized in Table 1.
Unveiling the underpinnings of various non-conventional ELISA variants: a review article
Published in Expert Review of Molecular Diagnostics, 2022
Rajesh Ahirwar, Akanksha Bhattacharya, Saroj Kumar
Over the past few decades, advances in immobilization chemistry, engineered antibodies and ultrasensitive enzyme substrates, nanomaterials-based signal enhancement strategies, and simplified signal detection procedures have enabled to achieve improved analytical performance while also sufficing the drawbacks of conventional microtiter-based ELISA protocol [21–25]. Advances in microfluidic techniques have led to the development of integrated microfluidic ELISA systems that are benefitted from the high surface-to-volume ratio, small sample consumption, and structural simplicity [26,27]. Practical feasibility of microfuidic ELISA platforms has been demonstrated on PDMS chip, polystyrene chip, glass capillaries, and centrifugal disc based systems [28]. Use of patterned cellulose (paper) as solid surface to carry out the antigen-antibody reaction and smart phones or scanner to capture the colored end product allows substantial cut down in the cost of ELISA assay [4,29]. Paper-ELISA carried out on paper microzone plates having same layout as plastic 96-well plates, with each zone requiring only 3 mL reagents, and the entire procedure completing in less than one hour opens opportunities for rapid, cost-effective, and point-of-care ELISA formats [30].
Organ-on-a-chip technologies that can transform ophthalmic drug discovery and disease modeling
Published in Expert Opinion on Drug Discovery, 2019
Jasmin C. Haderspeck, Johanna Chuchuy, Stefan Kustermann, Stefan Liebau, Peter Loskill
Su et al. developed a microfluidic chip to analyze retinal synaptic regeneration, which they called the retinal synaptic regeneration chip (RSR-chip) [74]. The RSR-chip contained two micro-chambers linked by multiple arrays of microchannels (see Figure 3(f)). Two cell populations derived from murine precursor cells were seeded separately into micro-chambers and after 3 days, axons emerged that grew from one population towards the other. Formation of retinal synaptic connections in response to secreted cytokines could be detected in the RSR-chip. Moreover, an oriented network of synapses was generated, and an automated method enabled simple analysis and enumeration of synaptic connectivity. The functionality of the chip was validated by testing the system with different amounts of glycine, which acts as an inhibitory transmitter in the retina. Furthermore, the electrophysiological activity of the retina could be monitored via immunodetection of pERK (phosphorylated extracellular-related kinase). The main purposes of this chip were to study the mechanism of synaptogenesis and to screen factors supporting retinal regeneration.