Role of detergents and nuclease inhibitors in the extraction of RNA from eukaryotic cells in complex matrices

Holohan, Cian; Feely, Nathan; Curran, Gerard; G, Lee

doi:10.1039/d2nr02850f

Cited by 3 publications

(4 citation statements)

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“…SAv NPs were prepared using different sizes of silica coated iron oxide (Fe 3 O 4 ) core–shell high magnetization NPs. [ 18–21 ] These silica coated NPs were amine functionalized by condensation deposition of 3‐aminopropyl triethoxysilane from an ethanolic solution followed by conversion of the terminal amine group to a carboxylic acid through a ring opening reaction with succinic anhydride. The immobilization of SAv was achieved though EDC/NHS activation of the surface carboxylic acid groups followed by incubation with SAv (see Figure A).…”

Section: Resultsmentioning

confidence: 99%

Targeting Mucin Protein Enables Rapid and Efficient Ovarian Cancer Cell Capture: Role of Nanoparticle Properties in Efficient Capture and Culture

Feely

Wdowicz

Chevalier³

et al. 2023

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The development of specific and sensitive immunomagnetic cell separation nanotechnologies is central to enhancing the diagnostic relevance of circulating tumor cells (CTCs) and improving cancer patient outcomes. The limited number of specific biomarkers used to enrich a phenotypically diverse set of CTCs from liquid biopsies has limited CTC yields and purity. The ultra‐high molecular weight mucin, mucin16 (MUC16) is shown to physically shield key membrane proteins responsible for activating immune responses against ovarian cancer cells and may interfere with the binding of magnetic nanoparticles to popular immunomagnetic cell capture antigens. MUC16 is expressed in ≈90% of ovarian cancers and is almost universal in High Grade Serous Epithelial Ovarian Cancer. This work demonstrates that cell bound MUC16 is an effective target for rapid immunomagnetic extraction of expressor cells with near quantitative yield, high purity and viability from serum. The results provide a mechanistic insight into the effects of nanoparticle physical properties and immunomagnetic labeling on the efficiency of immunomagnetic cell isolation. The growth of these cells has also been studied after separation, demonstrating that nanoparticle size impacts cell‐particle behavior and growth rate. These results present the successful isolation of “masked” CTCs enabling new strategies for the detection of cancer recurrence and select and monitor chemotherapy.

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Section: Resultsmentioning

confidence: 99%

Targeting Mucin Protein Enables Rapid and Efficient Ovarian Cancer Cell Capture: Role of Nanoparticle Properties in Efficient Capture and Culture

Feely

Wdowicz

Chevalier³

et al. 2023

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“…However, the use of cold protease to dissociate tissues at low temperatures has been demonstrated to enhance cellular activity and significantly reduce gene artefact expression [ 33 ]. To further preserve RNA integrity, it is viable to add nuclease inhibitors, such as SUPERaseIn and EDTA, to the lysis buffer [ 39 , 40 ]. Additionally, under high-fat conditions, the gene expression of liver cells may be significantly altered, resulting in the up- or downregulation of certain genes related to fat metabolism [ 41 , 42 ].…”

Section: Technologymentioning

confidence: 99%

Liver in infections: a single-cell and spatial transcriptomics perspective

Zhang

Zhong

et al. 2023

J Biomed Sci

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The liver is an immune organ that plays a vital role in the detection, capture, and clearance of pathogens and foreign antigens that invade the human body. During acute and chronic infections, the liver transforms from a tolerant to an active immune state. The defence mechanism of the liver mainly depends on a complicated network of intrahepatic and translocated immune cells and non-immune cells. Therefore, a comprehensive liver cell atlas in both healthy and diseased states is needed for new therapeutic target development and disease intervention improvement. With the development of high-throughput single-cell technology, we can now decipher heterogeneity, differentiation, and intercellular communication at the single-cell level in sophisticated organs and complicated diseases. In this concise review, we aimed to summarise the advancement of emerging high-throughput single-cell technologies and re-define our understanding of liver function towards infections, including hepatitis B virus, hepatitis C virus, Plasmodium, schistosomiasis, endotoxemia, and corona virus disease 2019 (COVID-19). We also unravel previously unknown pathogenic pathways and disease mechanisms for the development of new therapeutic targets. As high-throughput single-cell technologies mature, their integration into spatial transcriptomics, multiomics, and clinical data analysis will aid in patient stratification and in developing effective treatment plans for patients with or without liver injury due to infectious diseases.

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“…Yi et al [18] and Yi et al [19] then reported the use of non-ionic surfactant, Triton X-100, for reverse micelle formation. Nevertheless, due to the presence of the aromatic group and poor biodegradability of Triton X-100, it has been regarded as an environmentally unfriendly surfactant which is forbidden, especially in European countries [20,21].…”

Section: Introductionmentioning

confidence: 99%

“…Our previous works focused on the use of non-ionic polyethylene glycol (PEG)based [22] and alkyl polyglucoside (APG)-based [23] surfactants and rhamnolipid biosurfactant [24] for reverse micellar dyeing of cotton fabric in a different non-aqueous solvent medium. Compared with poly(ethylene glycol) (12) tridecyl ether (PEG-12), which is a mixture of C 11 to C 14 iso-alkyl ethers, Tergitol type 15-S-12 (T15S12) is a secondary alcohol ethoxylate (SAE) with distinctive branched hydrophobic tails [25], and it is readily biodegradable and specially created to meet the "designed to degrade" principle [21,26]. To the best of our knowledge, using T15S12 SAE-based biodegradable non-ionic surfactant (HLB value of 14.5) as a dye carrier for reverse micellar dyeing of cotton fabric is unknown and has not yet been explored and found in the literature.…”

Section: Introductionmentioning

confidence: 99%

Reverse Micellar Dyeing of Cotton Fabric with Reactive Dye Using Biodegradable Non-Ionic Surfactant as Nanoscale Carrier: An Optimisation Study by One-Factor-at-One-Time Approach

Tang,

Jin,

Lee

et al. 2023

Polymers

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This study investigates the feasibility of using biodegradable secondary alcohol ethoxylate (SAE) non-ionic surfactant as a building block for the formation of reverse micelles, functioning as reactive dye carriers for the dyeing of cotton fabric in non-aqueous octane medium. Ten dyeing parameters were optimised, by a one-factor-at-a-time approach, namely: (i) effect of colour fixation agent; (ii) surfactant-to-water mole ratio; (iii) surfactant-to-co-surfactant mole ratio; (iv) volume of soda ash; (v) volume of dye; (vi) solvent-to-cotton ratio; (vii) dyeing temperature; (viii) dyeing time; (ix) fixation time; (x) soda-ash-to-cotton ratio. The colour properties, fastness properties and physical properties of SAE-dyed samples were experimentally compared with the conventional water-dyed samples. The optimised condition was found when SAE samples were dyed as follows: (a) 1:20 surfactant-to-water ratio; (b) 1:8 surfactant-to-co-surfactant ratio; (c) 10:1 solvent ratio; (d) 40 min dyeing time; (e) 60 min fixation time; and (f) 70 °C dyeing and fixation temperature. The results showed that SAE-dyed samples have better colour strength, lower reflectance percentage and comparable levelness, fastness and physical properties than that of water-dyed samples. SEM images revealed that the dyed cotton fibres had no severe surface damage caused by an SAE-based reverse micellar dyeing system. The TEM image depicts that the reverse micelle was of nanoscale, spherical-shaped and had a core–shell structure, validating the presence of reverse micelle as a reactive dye carrier and the potential of an SAE-based reverse micellar system for dyeing of cotton fabrics.

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Role of detergents and nuclease inhibitors in the extraction of RNA from eukaryotic cells in complex matrices

Abstract: The potential for liquid biopsy samples to be used in place of more invasive tissue biopsy for the monitoring of patient tumours has become more prevalent in recent years. To...

Cited by 3 publications

References 31 publications

Targeting Mucin Protein Enables Rapid and Efficient Ovarian Cancer Cell Capture: Role of Nanoparticle Properties in Efficient Capture and Culture

Targeting Mucin Protein Enables Rapid and Efficient Ovarian Cancer Cell Capture: Role of Nanoparticle Properties in Efficient Capture and Culture

Liver in infections: a single-cell and spatial transcriptomics perspective

Reverse Micellar Dyeing of Cotton Fabric with Reactive Dye Using Biodegradable Non-Ionic Surfactant as Nanoscale Carrier: An Optimisation Study by One-Factor-at-One-Time Approach

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