scholarly journals Quantitative Detection of Cell Activity by Measuring the Fluctuation of Intracellular Motility

2019 ◽  
Author(s):  
Morito Sakuma ◽  
Yuichi Kondo ◽  
Hideo Higuchi
Keyword(s):  
Intensity Fluctuation ◽  
Cell Activity ◽  
Ips Cells ◽  
Intensity Change ◽  
Quantitative Detection ◽  
Oxygen Species ◽  
General Phenomenon ◽  
Transport Vesicles ◽  
Cytotoxic Molecules ◽  
Intracellular Motility

AbstractThe measurement of cell activity changes during damage is important to understand the process of cell death and evaluate the effect of medicines. To evaluate cell activity generally, we extended the method of intensity fluctuation in which intensity change in the pixel induced by the movement of organelles was calculated. Cancer, endothelial and iPS cells were damaged by reactive oxygen species (ROS) generated by a fluorescent dye (IR700), hydrogen peroxide, and ultraviolet light. The intensity fluctuation in damaged cells gradually decreased independent of the kind of cell, indicating that the decrease in the fluctuation is a general phenomenon in damaged cells. The rupture of vesicles and mitochondria in the cells were observed upon ROS production. The motility of purified kinesin and dynein which transport vesicles and organelles was inhibited by ROS. These suggest that ROS and cytotoxic molecules spreading from ruptured organelles contribute to the reduction in cell activity which brings about the decrease in the motility and intensity fluctuation of organelles driven by kinesin and dynein.

scholarly journals Novel Anthraquinone Compounds Inhibit Colon Cancer Cell Proliferation via the Reactive Oxygen Species/JNK Pathway

Molecules ◽  
2020 ◽  
Vol 25 (7) ◽  
pp. 1672 ◽  
Author(s):  
Yuying Li ◽  
Fang Guo ◽  
Yingying Guan ◽  
Tinggui Chen ◽  
Kaiqing Ma ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Colon Cancer ◽  
Cancer Cell ◽  
Human Cancer ◽  
Hct116 Cell ◽  
Cell Activity ◽  
Similarity Index ◽  
Reactive Oxygen ◽  
Field Analysis ◽  
Oxygen Species

A series of amide anthraquinone derivatives, an important component of some traditional Chinese medicines, were structurally modified and the resulting antitumor activities were evaluated. The compounds showed potent anti-proliferative activities against eight human cancer cell lines, with no noticeable cytotoxicity towards normal cells. Among the candidate compounds, 1-nitro-2-acyl anthraquinone-leucine (8a) showed the greatest inhibition of HCT116 cell activity with an IC50 of 17.80 μg/mL. In addition, a correlation model was established in a three-dimensional quantitative structure-activity relationship (3D-QSAR) study using Comparative Molecular Field Analysis (CoMFA) and comparative molecular similarity index analysis (CoMSIA). Moreover, compound 8a effectively killed tumor cells by reactive oxygen species (ROS)-JNK activation, causing an increase in ROS levels, JNK phosphorylation, and mitochondrial stress. Cytochrome c was then released into cytoplasm, which, in turn activated the cysteine protease pathway and ultimately induced tumor cell apoptosis, suggesting a potential use of this compound for colon cancer treatment.

scholarly journals POSSIBLE PREVENTION OF REACTIVE OXYGEN SPECIES INDUCED HUMAN TRABECULAR MESHWORK CELL DAMAGE BY RESVERATROL AND ASCORBIC ACID.

2019 ◽  
Vol 26 (07) ◽  
pp. 1036-1041
Author(s):  
Muhammad Yaqoob Shahani ◽  
Umbreen Bano ◽  
Shazia Begum Shahani ◽  
Pashmina Shaikh ◽  
Sameena Gul Memon ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Ascorbic Acid ◽  
Cell Viability ◽  
Trabecular Meshwork ◽  
Cell Metabolism ◽  
Cell Activity ◽  
Reactive Oxygen ◽  
Research Centre ◽  
Oxygen Species ◽  
Trabecular Meshwork Cells

Objectives: To analyze the antioxidant activity of Resveratrol and Ascorbic Acid against hydrogen peroxide (an oxidant) mediated cell injury of human trabecular meshwork cells. Study Design: Experimental study. Setting: Molecular Biology Laboratory at Medical Research Centre, Liaquat University of Medical & Health Sciences, Jamshoro. Period: Six months. Materials and Methods: Human Trabecular Meshwork cells were purchased from ScienCell Research Laboratories, USA. TM cell metabolism, TM cell viability and Reactive oxygen species were detected by standard methods in co- and pre- treated TM cells. Results: A significant reduction in TM cell metabolism was observed approximating 61% at 1.0 mM H2O2 compared to Ascorbate – 99% and Resveratrol 99% (p=0.0001). Resveratrol was more effective than Ascorbate even at 4.0 mM H2O2, the TM cell activity was noted 76%. Compared to H2O2- treated TM cells, resveratrol improved mitochondrial function upto 4.0 mM H2O2 (76%). Compared to co-treatment, the pretreatment shows similar results except at 4.0 mM H2O2. At 4.0 mM H2O2 the pre-treat TM cell metabolic activity was found as 11%, 31% and 47% compared to co-treat as 9%, 31% and 76% in controls, ascorbate and resveratrol groups respectively (p<0.05). Resveratrol shows significant decrease in viability was seen in controls compared to Ascorbate and Resveratrol groups. Cell viability showed statistically significant differences at 2.0 and 4.0 mM H2O2 compared to controls (P=0.0001). For reactive oxygen species (ROS), cells were incubated and with Ascorbate and Resveratrol for 24 hours and TM cells were treated with 0.0mM, 0.5 mM, 1.0 mM, 2.0mM and 4.0mM H2O2. Significant decrease in ROS was noted by Resveratrol compared to Ascorbate. Conclusions: Resveratrol and Ascorbate may prove useful in preventing and delaying the glaucoma, and timely institution of these anti – oxidants may help maintain trabecular meshwork functions and prevent visual loss.

scholarly journals Preprotection of Tea Polysaccharides with Different Molecular Weights Can Reduce the Adhesion between Renal Epithelial Cells and Nano-Calcium Oxalate Crystals

2020 ◽  
Vol 2020 ◽  
pp. 1-13
Author(s):  
Yao-Wang Zhao ◽  
Li Liu ◽  
Chuang-Ye Li ◽  
Hui Zhang ◽  
Xin-Yuan Sun ◽  
...  
Keyword(s):  
Calcium Oxalate ◽  
Cell Activity ◽  
Calcium Oxalate Monohydrate ◽  
Oxygen Species ◽  
Calcium Oxalate Crystals ◽  
Renal Epithelial Cells ◽  
Molecular Weights ◽  
Before And After ◽  
Tea Polysaccharides ◽  
Crystal Adhesion

Crystal adhesion is an important link in the formation of kidney stones. This study investigated and compared the adhesion differences between nano-calcium oxalate monohydrate (COM) and human renal proximal tubule epithelial (HK-2) cells before and after treatment with tea polysaccharides (TPSs) TPS0, TPS1, TPS2, and TPS3 with molecular weights of 10.88, 8.16, 4.82, and 2.31 kDa, respectively. TPS treatment effectively reduced the damage of COM to HK-2 cells, thereby resulting in increased cell activity, decreased release of lactate dehydrogenase, cell morphology recovery, decreased level of reactive oxygen species, increased mitochondrial membrane potential, increased lysosomal integrity, decreased expression of adhesion molecule osteopontin and eversion of phosphatidylserine, and decreased crystal adhesion. Among the TPSs, TPS2 with moderate molecular weight had the best protective effect on cells and the strongest effect on the inhibition of crystal adhesion. Thus, TPS2 may be a potential anticalculus drug.

Mechanism of Biological Compatibility of Water-Soluble Yb3+, Er3+ Codoped NaYF4 Nanoparticles

2016 ◽  
Vol 16 (4) ◽  
pp. 3653-3658 ◽  
Author(s):  
Chen Liu ◽  
Huan Chen ◽  
Shouying Li ◽  
Hong Xu ◽  
Dan Zhao
Keyword(s):  
Cell Viability ◽  
Cell Activity ◽  
Suppressor Cells ◽  
Water Soluble ◽  
Oxygen Species ◽  
Cell Cycles ◽  
Luminescent Probes ◽  
Biological Compatibility ◽  
Apoptosis Detection ◽  
Induced Apoptosis

Water soluble NaYF4 nanocrystals codoped with 20 mol% Yb3+, 2 mol% Er3+ were prepared by a facile solvothermal approach using polyvinylpyrrolidone (PVP) as a surfactant. As a potential material for luminescent probes, in votroeffects of upconversion nanoparticles (UCNPs) on human aenocarcinoma (SGC-7901) cells with different concentrations were observed. These effects range from cell viability, mitochondrial membrane potential (MMP), reactive oxygen species (ROS) production, AnnexinV-FITC–propidiumiodide (PI) apoptosis detection, and cell cycles. Our results demonstrated that the cells treated with UCNPs showed a decrease in cell viability accompanied the decreased MMP and the release of ROS. When treated with 400 μg/mL UCNPs, AnnexinV-FITC–PI apoptosis detection showed the UCNPs induced apoptosis, the cell cycle indicated the UCNPs suppressor cells in the G1 phase obviously, thereby reducing cell activity.

scholarly journals Detection of Vascular Reactive Oxygen Species in Experimental Atherosclerosis by High-Resolution Near-Infrared Fluorescence Imaging Using VCAM-1-Targeted Liposomes Entrapping a Fluorogenic Redox-Sensitive Probe

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
Simona-Adriana Manea ◽  
Mihaela-Loredana Vlad ◽  
Daniela Rebleanu ◽  
Alexandra-Gela Lazar ◽  
Ioana Madalina Fenyo ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
High Resolution ◽  
Near Infrared ◽  
Reactive Oxygen ◽  
Experimental Models ◽  
Fluorescent Imaging ◽  
Quantitative Detection ◽  
Oxygen Species ◽  
Near Infrared Fluorescence ◽  
Models Of Disease

Excessive production of reactive oxygen species (ROS) and the ensuing oxidative stress are instrumental in all phases of atherosclerosis. Despite the major achievements in understanding the regulatory pathways and molecular sources of ROS in the vasculature, the specific detection and quantification of ROS in experimental models of disease remain a challenge. We aimed to develop a reliable and straightforward imaging procedure to interrogate the ROS overproduction in the vasculature and in various organs/tissues in atherosclerosis. To this purpose, the cell-impermeant ROS Brite™ 700 (RB700) probe that produces bright near-infrared fluorescence upon ROS oxidation was encapsulated into VCAM-1-targeted, sterically stabilized liposomes (VLp). Cultured human endothelial cells (EC) and macrophages (Mac) were used for in vitro experiments. C57BL6/J and ApoE-/- mice were randomized to receive normal or high-fat, cholesterol-rich diet for 10 or 32 weeks. The mice received a retroorbital injection with fluorescent tagged VLp incorporating RB700 (VLp-RB700). After two hours, the specific signals of the oxidized RB700 and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-DSPE), inserted into liposome bilayers, were measured ex vivo in the mouse aorta and various organs by high-resolution fluorescent imaging. VLp-RB700 was efficiently taken up by cultured human EC and Mac, as confirmed by fluorescence microscopy and spectrofluorimetry. After systemic administration in atherosclerotic ApoE-/- mice, VLp-RB700 were efficiently concentrated at the sites of aortic lesions, as indicated by the augmented NBD fluorescence. Significant increases in oxidized RB700 signal were detected in the aorta and in the liver and kidney of atherosclerotic ApoE-/- mice. RB700 encapsulation into sterically stabilized VCAM-1-sensitive Lp could be a novel strategy for the qualitative and quantitative detection of ROS in the vasculature and various organs and tissues in animal models of disease. The accurate and precise detection of ROS in experimental models of disease could ease the translation of the results to human pathologies.

Levels of Monocyte Reactive Oxygen Species Are Associated with Reduced Natural Killer Cell Activity in Major Depressive Disorder

2001 ◽  
Vol 44 (1) ◽  
pp. 1-6 ◽  
Author(s):  
Matthew G. Frank ◽  
Shelton E. Hendricks ◽  
Diane Bessette ◽  
Donald R. Johnson ◽  
Julie L. Wieseler Frank ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Major Depressive Disorder ◽  
Natural Killer Cell ◽  
Depressive Disorder ◽  
Natural Killer Cell Activity ◽  
Killer Cell ◽  
Cell Activity ◽  
Oxygen Species ◽  
Major Depressive ◽  
Killer Cell Activity

Bionutrition and Oral Cancer in Humans

1995 ◽  
Vol 6 (1) ◽  
pp. 5-17 ◽  
Author(s):  
C.O. Enwonwu ◽  
V.I. Meeks
Keyword(s):  
Free Radicals ◽  
Oral Cancer ◽  
T Cell Activation ◽  
Tobacco Smoking ◽  
Cell Activity ◽  
Dietary Factors ◽  
Free Radical Scavengers ◽  
Oxygen Species ◽  
Incidence And Mortality ◽  
Anti Oxidant

Tobacco (smoking and smokeless) use and excessive consumption of alcohol are considered the main risk factors for oral cancer (ICD9 140-149). Conspicuous national and international variations in oral cancer incidence and mortality rates, as well as observations in migrant populations, raise the possibility that diet and nutritional status could be an important etiologic factor in oral carcinogenesis. As shown in this report, abuse of alcohol and tobacco has serious nutritional implications for the host, and generates increased production of reactive free radicals as well as eliciting immunosuppression. Maintenance of optimal competence of the immune system is critical for cancer surveillance. Active oxygen species and other reactive free radicals mediate phenotypic and genotypic alterations that lead from mutation to neoplasia. Consequently, the most widely used chemopreventive agents against oral cancer (e.g., vitamins A, E, C, and β-carotene) are anti-oxidants/free radical scavengers. These anti-oxidants, both natural and synthetic, neutralize metabolic products (including reactive oxygen species), interfere with activation of procarcinogens, prevent binding of carcinogens to DNA, inhibit chromosome aberrations, restrain replication of the transformed cell, suppress actions of cancer promoters, and may even induce regression of precancerous oral lesions such as leukoplakia and erythroplakia. Malnutrition is characterized by marked tissue depletion of anti-oxidant nutrients, including GSH (y-glutamyl-cysteinyl-glycine), a key cellular anti-oxidant as well as a modulator of T-cell activation. GSH or its precursor cysteine inhibits activation of the nuclear transcription factor kB (NFkB), and has been shown to be protective against chemically induced oral cancer and leukoplakia. Alcohol-, tobacco-, and/or malnutrition-induced immunosuppression promotes impaired salivary gland function and oral mucosal immunity, a prominent reduction in the number of helper CD4 cells with less marked changes in number of suppressor T-cells, and depressed NK cell activity, among others. These suggest a breakdown in the capacity of the malnourished to mount effective tumor surveillance. This review article underscores the compounding but important roles of nutritional/dietary factors in the long-established causal link between abuse of alcohol and tobacco (smoking and smokeless) and oral cancer.

General redox environment and carotid body chemoreceptor function

2009 ◽  
Vol 296 (3) ◽  
pp. C620-C631 ◽  
Author(s):  
Maria Teresa Agapito ◽  
Gloria Sanz-Alfayate ◽  
Angela Gomez-Niño ◽  
Constancio Gonzalez ◽  
Ana Obeso
Keyword(s):  
Oxidative Stress ◽  
Reactive Oxygen Species ◽  
Carotid Body ◽  
Cell Activity ◽  
Oxygen Species ◽  
Oxidizing Agents ◽  
Redox Environment ◽  
Chemoreceptor Cells ◽  
Glutathione Redox

Carotid body (CB) chemoreceptor cells detect physiological levels of hypoxia and generate a hyperventilation, homeostatic in nature, aimed to minimize the deleterious effects of hypoxia. Intimate mechanisms involved in oxygen sensing in chemoreceptor cells remain largely unknown, but reactive oxygen species (ROS) had been proposed as mediators of this process. We have determined glutathione levels and calculated glutathione redox potential ( EGSH; indicator of the general redox environment of cells) in rat diaphragms incubated in the presence of oxidizing agents of two types: nonpermeating and permeating through cell membranes; in the latter group, unspecific oxidants and inhibitors of ROS-disposing enzymes were used. Selected concentrations of oxidizing agents were tested for their ability to modify the normoxic and hypoxic activity of chemoreceptor cells measured in vitro as their rate of release of neurotransmitters. Results evidence variable relationships between EGSHand the activity of chemoreceptor cells. The independence of chemoreceptor cell activity from the EGSHwould imply that the ability of the CB to play its homeostatic role is largely preserved in any pathological or toxicological contingency causing oxidative stress. Consistent with this suggestion, it was also found that CB-mediated hypoxic hyperventilation was not altered by treatment of intact animals with agents that markedly decreased the EGSHin all tissues assayed.

NAC Improves the Differentiation of IPS Cells Into Hematopoietic Progenitors

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 2349-2349
Author(s):  
Ina Berniakovich ◽  
Leopoldo Laricchia-Robbio ◽  
Juan Carlos Izpisua
Keyword(s):  
Reactive Oxygen Species ◽  
Stress Resistance ◽  
Mitochondrial Membrane ◽  
Reactive Oxygen ◽  
Ips Cells ◽  
Oxygen Species ◽  
Cd34 Cells ◽  
Human Ips Cells

Abstract Abstract 2349 In vitro differentiation of human induced pluripotent stem (iPS) cells is a new way to obtain donor material for blood transplantation. However, this is a long process in the course of which cells are exposed to alien environmental factors, capable to change cellular properties and decrease cellular viability. Indeed, cells in vitro are exposed to mechanical stress, artificial growth surface, unnatural gas composition etc. This fact could partially explain why cells obtained during directed iPS cells differentiation have different qualities in comparison with the analogous population from in vivo. It is supposed that oxidative stress is the major underlying mechanism of negative influence of various in vitro environmental factors on cells. N-acetylcysteine (NAC) is a powerful antioxidant. Due to free radical scavenging ability, the principal function of NAC is rendered to the inhibition of cellular damage and cell death in response to reactive oxygen species. Cytoprotective function of NAC is well demonstrated both in vitro and in vivo. We have established a protocol to obtain hematopoietic stem cell-like cells from human iPS cells with a pick of their production at three weeks of differentiation. We analyzed how intracellular accumulation of reactive oxygen species and NO, as well as cell viability, apoptosis, stress resistance, mitochondrial membrane potential were changed during this process by comparing status of cultures at 1 and at 3 weeks of differentiation. We found that during differentiation cells progressively accumulated intracellular reactive oxygen species and increased production of NO. The level of apoptosis in culture was significantly higher at 3 weeks of differentiation than at 1 week. Cell viability, on the contrary, decreased from 1 week till 3 weeks of differentiation. Stress resistance quantified through the amount of cells resistant to the H2O2 treatment was also decreased in 3 weeks old cultures. We also demonstrated that during in vitro culture the mitochondrial membrane potential of the cells under basal conditions and upon stimulation with carbonyl cyanide m-chlorophenylhydrazone was decreased at 3 weeks of differentiation in comparison with that at 1 week. All these phenomena were reversed by NAC supplementation. Remarkably, NAC administration also improved the hematopoietic differentiation of human iPS cells in terms of production of CD34, CD45, CD43 positive cells, that showed normal functionality in colony forming unit assay. CD34+ cells obtained from NAC treated cultures also increased their migration towards SDF1, therefore showing an increased ability of our CD34+ cells to home into bone marrow. Our results suggest that supplementation with NAC is beneficial for the improvement of hematopoietic differentiation of human iPS cells. Disclosures: No relevant conflicts of interest to declare.

In vitro modelling of familial amyloidotic polyneuropathy allows quantitative detection of transthyretin amyloid fibril-like structures in hepatic derivatives of patient-specific induced pluripotent stem cells

2017 ◽  
Vol 398 (8) ◽  
pp. 939-954 ◽  
Author(s):  
Jeannine Hoepfner ◽  
Mandy Kleinsorge ◽  
Oliver Papp ◽  
Susanne Alfken ◽  
Robin Heiringhoff ◽  
...  
Keyword(s):  
Amyloid Fibril ◽  
Ips Cells ◽  
Fibril Formation ◽  
Thioflavin T ◽  
Familial Amyloidotic Polyneuropathy ◽  
Patient Specific ◽  
Quantitative Detection ◽  
Healthy Control ◽  
Induced Pluripotent

Abstract The transthyretin protein is thermodynamically destabilised by mutations in the transthyretin gene, promoting the formation of amyloid fibrils in various tissues. Consequently, impaired autonomic organ function is observed in patients suffering from transthyretin-related familial amyloidotic polyneuropathy (FAP). The influence of individual genetic backgrounds on fibril formation as a potential cause of genotype-phenotype variations needs to be investigated in order to ensure efficient patient-specific therapies. We reprogrammed FAP patient fibroblasts to induced pluripotent stem (iPS) cells and differentiated these cells into transthyretin-expressing hepatocyte-like cells (HLCs). HLCs differentiated from FAP iPS cells and healthy control iPS cells secreted the transthyretin protein in similar concentrations. Mass spectrometry revealed the presence of mutant transthyretin protein in FAP HLC supernatants. In comparison to healthy control iPS cells, we demonstrated the formation of transthyretin amyloid fibril-like structures in FAP HLC supernatants using the amyloid-specific dyes Congo red and thioflavin T. These dyes were also applicable for the quantitative determination of in vitro formed transthyretin fibril-like structures. Moreover, we confirmed the inhibition of fibril formation by the TTR kinetic stabiliser diclofenac. Thioflavin T fluorescence intensity measurements even allowed the quantification of amyloid fibril-like structures in 96-well plate formats as a prerequisite for patient-specific drug screening approaches.

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