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Natural Product Compounds from Plants in Neurodegenerative Diseases
Published in Namrita Lall, Medicinal Plants for Cosmetics, Health and Diseases, 2022
Priya Darshani, Md TanjimAlam, Prem P. Tripathi, V.S. Pragadheesh
Korean red ginseng was shown to prevent dopaminergic neuron cell death by decreasing the expression of cyclin-dependent kinase 5 (Cdk5) in the substantia nigra and striatum. In the cellular model on rat PC12 cells, it was shown that Korean red ginseng treatment prevented 1-methyl-4-phenylpyridinium ion- (MPP+-)–induced cell death. Decreased cell viability and apoptosis were consequently reported. Another plant, Uncaria rhynchophylla (Miq.) Miq. ex Havil., reduced apoptosis and reactive oxygen species (ROS) generation, therefore exhibiting anti-Parkinson’s activity (Sairazi and Sirajudeen, 2020). Flavonoids such as baicalein, hesperidin, and quercetin were reported to increase dopamine and serotonin levels in the striatum and downregulate oxidative stress and the astroglial response, thus highlighting their anti-neurodegenerative roles in PD mice models (Corona, 2018). Administration of monosialotetrahexosylganglioside (GM1) can alleviate the effects of various neurodegenerative processes. GM1-deficient animals showed Parkinson-like symptoms, suggesting the potential therapeutic role of GM1 in PD treatment (Wang et al., 2019).
Flow Cytometric Analysis of Human Bone Marrow
Published in Adrian P. Gee, BONE MARROW PROCESSING and PURGING, 2020
James G. Bender, Dennis Van Epps
Another major concern with the use of any ex vivo manipulation of bone marrow is the loss of viability that may take place. Besides the importance of measuring the effect of any processing procedure on viability, it is also important to understand the problems that nonviable cells create in quantitating immunofluorescence, as these cells will bind antibodies nonspecifically. The approaches to assessing cell viability by flow cytometry can be divided into techniques for evaluating either nonfixed or fixed preparations for the presence of nonviable or damaged cells. In the case of fixed cells, assessment of viability refers to methods that identify cells that were nonviable before fixation. In general, the best results for evaluating nonviable populations are obtained using a multiparameter approach, in which the nonviable cells are resolved as a separate population and can then be quantitated and removed from subsequent analyses.
Serological Typing of HLA-A, -B, and -C Antigens
Published in M. Kam, Jeffrey L. Bidwell, Handbook of HLA TYPING TECHNIQUES, 2020
An expanded polystyrene box of similar dimensions to that given above is used (box 1). A larger expanded polystyrene box (box 2) is also required with sufficient dimensions to allow the circulating air space around box 1 (Figure 4). Place the rack of microcentrifuge tubes/ampoules containing the lymphocytes in box 1. Then place box 1 on the supporting rack inside box 2 containing liquid nitrogen. Replace the lid and place a small weight on it to prevent liquid nitrogen evaporation. Leave for 30 and 60 min for microcentrifuge tubes and ampoules, respectively. Transfer the vials to liquid nitrogen. Various box combinations should be tested to obtain the maximum cell recovery and cell viability.
Synthetic pyrethroids common metabolite 3-phenoxybenzoic acid induces caspase-3 and Bcl-2 mediated apoptosis in human hepatocyte cells
Published in Drug and Chemical Toxicology, 2022
Dilek Guvenc, Sinem Inal, Nilufer Kuruca, Sedat Gokmen, Tolga Guvenc
The cytotoxic effects of 3-PBA on various cell types have been demonstrated to be dose and time dependent in immunotoxicity and neurotoxicity studies (Romero et al.2012, Wang et al.2017, He et al.2018). Research on the toxic effects of 3-PBA, including hepatotoxic effects, is still limited. In the present study, the aim of the investigation of 3-PBA induced apoptosis in HepG2 cells was to progress the understanding of the mechanisms of 3-PBA toxicity. Our results demonstrated a significant, dose-dependent decrease in cell viability. Immunotoxicity studies have reported that β-CYP, and its metabolite 3-PBA, reduce cell viability in mouse macrophage cells and human promyelocytic leukemia cells, both time and dose-dependent (Wang et al.2017, He et al.2018).
Evaluation of PEG-b-polycarbonates self-assemblies containing azobenzene or coumarin moieties as nanocarriers using paclitaxel as a model hydrophobic drug
Published in Journal of Microencapsulation, 2022
Alejandro Roche, Violeta Morcuende-Ventura, Rosa M. Tejedor, Luis Oriol, Olga Abian, Milagros Piñol
Unloaded and PTX-loaded PEG2k-b-PC(AzoOMe) vesicles were evaluated for their cytotoxicity against HeLa cell line using CellTiter 96® Reagent for incubation times of 72h. To have comparable data, the polymer concentration was the same for both sets of experiments. i.e. either with unloaded or PTX-loaded vesicles. Besides, results were contrasted with those obtained with free-PTX at the same PTX concentration. The results of cell viability studies are collected in Figure 3. Cell viability was almost unaffected by incubation with unloaded PEG2k-b-PC(AzoOMe) vesicles as far as BC concentration was kept below 4μg mL‒1 (Figure 3a). However, at the same PTX concentration, PTX-loaded vesicles showed an enhanced cytotoxicity in comparison to free-PTX in all the range of concentrations studied (Figure 3b). Half cytotoxic concentration (IC50) defined as the PTX concentration that it is cytotoxic for half of the initial cell population (i.e. PTX concentration for which cell viability is 50%) was determined (Figure 3b). IC50 for free-PTX was 0.17±0.02nM (p<0.05) whereas the value for PTX-loaded vesicles was about eight times lower (Figure 3b). Therefore, PTX cytotoxicity against HeLa cells was enhanced by encapsulation into the self-assemblies so a lower concentration of PTX should be necessary if encapsulated to achieve the same effect than free PTX.
In vitro toxicity evaluation of lomefloxacin-loaded MCM-41 mesoporous silica nanoparticles
Published in Drug and Chemical Toxicology, 2021
Virginia Tzankova, Denitsa Aluani, Yordan Yordanov, Massimo Valoti, Maria Frosini, Ivanka Spassova, Daniela Kovacheva, Borislav Tzankov
Human hepatoma cells Hep G2, L929 mouse fibroblast cells and endothelial EA.hy926 cells were obtained from Sigma Aldrich (ECACC cell lines). Hep G2 cells were kept in culture at 37 °C in a humidified atmosphere of 5% CO2 in culture medium DMEM (Dulbecco's Modified Eagle's Medium, Lonza, Bazel, CH), supplemented with 20% fetal bovine serum (FBS) (Gibco BRL) at 10%, penicillin/streptomycin 100× (Euroclone, Devon, UK), Glutamax 100× (Invitrogen) and non-essential amino acids 100× (Invitrogen). L929 mouse fibroblast cells have been employed as an appropriate in vitro system for biocompatibility assessment (ISO 10993 2009). L929 cells were cultured in DMEM medium, supplemented with 10% fetal bovine serum (FBS) (Gibco BRL) and 10%, penicillin/streptomycin 100× (Euroclone, Devon, UK). EA.hy926 cells were grown at 5% CO2 in Dulbecco's modified Eagle's medium, supplemented with 10% fetal calf serum, 2 mM L-glutamine, 1 mM sodium pyruvate, 100 IU/ml penicillin, 100 μg/ml streptomycin. EA.hy926 cell culture medium was supplemented with 1 × HAT (5 mM hypoxanthine, 20 μM aminopterin, and 0.8 mM thymidine; 21060–017; Gibco). Cell viability was routinely measured using trypan blue exclusion test and cell populations with at least 90% viability were used for the experiments.