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Quercetin Bioflavonoids Derived from Phytomedicinal Compounds for Targeted Drug Delivery and Their Antioxidant Properties
Published in Parimelazhagan Thangaraj, Lucindo José Quintans Júnior, Nagamony Ponpandian, Nanophytomedicine, 2023
Selvaraj Rajesh Kumar, Nagamony Ponpandian
The quercetin-loaded silver nanoparticle shows an effective antioxidant property that reduced lipid oxidation and improved antibacterial activities. The quercetin-loaded poly(D, L-lactide-co-glycolide) (PLGA) nanoparticles with the size range of 100–150 nm were used as an antioxidant compound and enhanced antibacterial activity. Solid lipid nanoparticles loaded with quercetin protect retinal ganglion and cornea eye cells from oxidative damage by encapsulation that retains higher cytotoxicity and protects the eye from cancer attacks. Quercetin conjugated gold nanoparticles induce cell apoptosis in human breast cancer cells through epidermal growth factor receptor (EGFR)–mediated PI3K/Akt/GSK-3 β signalling by immunocytochemistry and immunoblotting. Quercetin loaded with the GO-PVP nanovehicle improves targeting efficiency, dissolution rate and induces more cell apoptosis in ovarian cells. Hence, the nanomaterials act as effective nanocarriers to quercetin for targeted drug delivery applications.
Immunochemistry of Cell Wall – a Tool for Evaluation of the Response of Plants to Changed Habitat
Published in Artur Dyczko, Andrzej M. Jagodziński, Gabriela Woźniak, Green Scenarios: Mining Industry Responses to Environmental Challenges of the Anthropocene Epoch, 2022
Katarzyna Sala, Kamila Godel-Jędrychowska, Ewa Kurczyńska
Immunochemistry (ICH) includes immunocytochemistry (ICC) which examines cells and immunohistochemistry (IHC) which examines the tissues. It is a method that allows for identification or localisation of specific antigens within cells or tissues. The method utilises an antigen-specific antibody which binds an antigen in a cell/tissue section, fluorochromes (at the light microscope level) and colloidal-gold particles (at the electron microscopy level) to visualize where the antibody was bound. ICH, nowadays, is a common laboratory technique that is used to visualize the presence of a specific antigen in cells/tissues by use of a specific primary antibody that binds to it. In a two-step reaction, the primary antibody allows for visualization of the antigen under a fluorescence microscope only when it is bound by a secondary antibody that has a conjugated fluorophore (Fig. 2). ICH helps to evaluate whether or not cells/tissues in a particular sample express the antigen in question. In cases where an immunopositive signal is found, we are able to determine which sub-cellular compartments are expressing the antigen.
Surface Chemistry for Cell Capture in Microfluidic Systems
Published in Iniewski Krzysztof, Integrated Microsystems, 2017
ShuQi Wang, Feng Xu, Alexander Chi Fai Ip, Mrudula Somu, Xiaohu Zhao, Altug Akay, Utkan Demirci
At the cellular level, an enzyme-based fluorescent detection system is commonly used to achieve a direct visualization of captured cancer cells [44]. This method employs a fluorescent dye-conjugated antibody that is specific for different cancer cell types, allowing for quick fluorescence detection. Immunocytochemistry staining can also be used to confirm capture cells by targeting cell surface biomarkers [17]. Other physical properties of CTCs and normal tissue cells are also often used for CTC detection [45]. One such change is that the cancer cells become softer and more deformable [46]. This unique feature allows the detection of human kidney carcinoma cells using impedimetric transducers by monitoring the volume increase of cancer cells under hyposmotic pressure [47]. Human kidney cancer cells will swell under hyposmotic stress compared to the unchanging normal cells [47]. This application allowed the detection of cancer cells from whole blood without prelabeling or capturing. Alternatively, the size differences between cancer and noncancer cells can also be used to detect CTCs, based on the measurement of conductance [39].
Derivation of induced pluripotent stem cell lines from New Zealand donors
Published in Journal of the Royal Society of New Zealand, 2022
Jin Kyo Oh, Aneta Przepiorski, Hao-Han Chang, Rachel C. Dodd, Veronika Sander, Brie Sorrenson, Jen-Hsing Shih, Jennifer A. Hollywood, Janak R. de Zoysa, Peter R. Shepherd, Alan J. Davidson, Teresa M. Holm
We used the integration-free Sendai virus system to reprogram dermal fibroblasts from two adult donors towards pluripotency. iPSC colonies emerged 3–4 weeks post-transduction and were picked to generate individual lines. We obtained multiple clones of the MANZ-2 and MANZ-4 lines, which could be stably passaged and expanded while exhibiting the morphology characteristic of human ESCs (Figure 1A). Six iPSC lines per donor were randomly selected for karyotyping. MANZ-2-2 and MANZ-4-37 lines showed no chromosome abnormalities and were selected for further characterisation (Figure 1B). STR analysis for 16 loci confirmed the genetic integrity of the MANZ lines (Supplementary Figure 3). Using qPCR, we measured that transcript levels of the pluripotency genes OCT4, NANOG and SOX2 were highly induced in both MANZ lines compared to the corresponding fibroblasts (Figure 1C). cMYC induction was less prominent, presumably due to a high base expression of cMYC in the corresponding fibroblasts (Waters et al. 1991). Sendai virus RNA was not detectable by RT–PCR (Supplementary Figure 4), indicating that our lines successfully maintained pluripotency without viral transgene expression. Immunocytochemistry confirmed the presence of the pluripotency cell-surface-antigen markers SSEA-4, TRA-1-60 and TRA-1-81 (Figure 1D), whereas the differentiation marker SSEA-1 was undetectable in both MANZ lines (not shown).
Decellularization and genipin crosslinking of amniotic membrane suitable for tissue engineering applications
Published in Journal of Biomaterials Science, Polymer Edition, 2018
Sarumathi Gobinathan, Siti Solehah Zainol, Siti Fatmah Azizi, Nabil Mohamad Iman, Rajasegaran Muniandy, Hanis Nazihah Hasmad, Mohd Reusmaazran bin Yusof, Salina Husain, Haslinda Abd Aziz, Yogeswaran Lokanathan
Immunocytochemistry of the extracellular matrix was performed only on nAM and dAM to study the ECM lost due to decellularization process. Cells on HAM were washed with DPBS and fixed with 4% paraformaldehyde for overnight (Sigma-Aldrich, USA), permeabilized for 20 minutes with 0.5% Triton X-100 solution (Sigma-Aldrich), and then blocked with 10% goat serum for 1 hour at 37°C. The cells were incubated mouse anti-collagen type 1, mouse anti-collagen type 3, and mouse anti-collagen type 4 (Abcam, USA) for overnight at 4°C. Next day, the cells were washed before being incubated with 1:300 diluted goat anti-mouse Alexa Fluor 594 (Invitrogen) for 1 hour at 37°C. The cells were then observed using Nikon Eclipse Ti fluorescence microscope (Nikon, Japan).
Genetic toxicity assessment using liver cell models: past, present, and future
Published in Journal of Toxicology and Environmental Health, Part B, 2020
Xiaoqing Guo, Ji-Eun Seo, Xilin Li, Nan Mei
The γH2AX assay has been used as a new approach for detecting DNA damage based upon the concept that DNA double-strand breaks induce a rapid phosphorylation of the carboxyl-tail of histone H2AX at Serine 139, i.e., γH2AX (Watters et al. 2009). H2AX phosphorylation is considered a more sensitive marker of DNA damage than currently available techniques (Toyooka, Ishihama, and Ibuki 2011). There are predominantly two techniques for detecting γH2AX, (1) measurement of γH2AX foci formation by immunocytochemistry or flow cytometry, and (2) quantification of γH2AX protein in fixed cultured cells using antibodies, specifically, the 96-well In Cell Western (ICW) assay.