scholarly journals Doxyl Nitroxide Spin Probes Can Modify Toxicity of Doxorubicin towards Fibroblast Cells

Molecules ◽  
2020 ◽  
Vol 25 (21) ◽  
pp. 5138
Author(s):  
Jan Czepas ◽  
Karolina Matczak ◽  
Aneta Koceva-Chyła ◽  
Bartłomiej Grobelski ◽  
Zofia Jóźwiak ◽  
...  
Keyword(s):  
Cell Death ◽  
Plasma Membrane ◽  
Membrane Fluidity ◽  
Biological Properties ◽  
Spin Probes ◽  
Trypan Blue Exclusion ◽  
Paramagnetic Resonance ◽  
Trypan Blue Exclusion Test ◽  
High Concentrations ◽  
Nitroxide Spin Probes

The biological properties of doxyl stearate nitroxides (DSs): 5-DS, Met-12-DS, and 16-DS, commonly used as spin probes, have not been explored in much detail so far. Furthermore, the influence of DSs on the cellular changes induced by the anticancer drug doxorubicin (DOX) has not yet been investigated. Therefore, we examined the cytotoxicity of DSs and their ability to induce cell death and to influence on fluidity and lipid peroxidation (LPO) in the plasma membrane of immortalised B14 fibroblasts, used as a model neoplastic cells, susceptible to DOX-induced changes. The influence of DSs on DOX toxicity was also investigated and compared with that of a natural reference antioxidant α-Tocopherol. By employing the trypan blue exclusion test and double fluorescent staining, we found a significant level of cytotoxicity for DSs and showed that their ability to induce apoptosis and modify plasma membrane fluidity (measured fluorimetrically) is more potent than for α-Tocopherol. The most cytotoxic nitroxide was 5-DS. The electron paramagnetic resonance (EPR) measurements revealed that 5-DS was reduced in B14 cells at the fastest and Met-12-DS at the slowest rate. In the presence of DOX, DSs were reduced slower than alone. The investigated compounds, administered with DOX, enhanced DOX-induced cell death and demonstrated concentration-dependent biphasic influence on membrane fluidity. A-Tocopherol showed weaker effects than DSs, regardless the mode of its application—alone or with DOX. High concentrations of α-Tocopherol and DSs decreased DOX-induced LPO. Substantial cytotoxicity of the DSs suggests that they should be used more carefully in the investigations performed on sensitive cells. Enhancement of DOX toxicity by DSs showed their potential to act as chemosensitizers of cancer cells to anthracycline chemotherapy.

Inherent Microporosity and Photostability of Fluoroacrylic Polymer Films Studied by Electron Paramagnetic Resonance of Nitroxide Spin Probes

2015 ◽  
Vol 46 (5) ◽  
pp. 523-540 ◽  
Author(s):  
A. M. Sheveleva ◽  
I. K. Shundrina ◽  
S. L. Veber ◽  
A. D. Buhtojarova ◽  
V. V. Russkih ◽  
...  
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Polymer Films ◽  
Spin Probes ◽  
Paramagnetic Resonance ◽  
Nitroxide Spin Probes ◽  
Electron Paramagnetic

Polygalae Radix Extract Protects Cultured Rat Granule Cells Against Damage Induced by NMDA

2004 ◽  
Vol 32 (04) ◽  
pp. 599-610 ◽  
Author(s):  
Hyun Joo Lee ◽  
Ju Yeon Ban ◽  
Sang Bum Koh ◽  
Nak Sul Seong ◽  
Kyung Sik Song ◽  
...  
Keyword(s):  
Cell Death ◽  
Granule Cells ◽  
Cell Damage ◽  
Glutamate Release ◽  
Neuronal Cell Death ◽  
Neuronal Cell ◽  
Inhibitory Effect ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Test

Polygalae Radix (PR) from Polygala tenuifolia (Polygalaceae) is traditionally used in China and Korea, as this herb has a sedative, anti-inflammatory and antibacterial agent. To extend our understanding of the pharmacological actions of PR in the CNS on the basis of its CNS inhibitory effect, the present study examined whether PR has the neuroprotective action against N-methyl-D-aspartate (NMDA)-induced cell death in primarily cultured rat cerebellar granule neurons. PR, over a concentration range of 0.05 to 5 μg/ml, inhibited NMDA (1 mM)-induced neuronal cell death, which was measured by a trypan blue exclusion test and a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. PR (0.5 μg/ml) inhibited glutamate release into medium induced by NMDA (1 mM), which was measured by HPLC. Pre-treatment of PR (0.5 μg/ml) inhibited NMDA (1 mM)-induced elevation of intracellular Ca 2+ concentration ([ Ca 2+] i ), which was measured by a fluorescent dye, Fura 2-AM, and generation of reactive oxygen species (ROS). These results suggest that PR prevents NMDA-induced neuronal cell damage in vitro.

Kinetics of cell death of frozen-thawed human embryonic stem cell colonies is reversibly slowed down by exposure to low temperature

Zygote ◽  
2006 ◽  
Vol 14 (4) ◽  
pp. 341-348 ◽  
Author(s):  
B.C. Heng ◽  
C.P. Ye ◽  
H. Liu ◽  
W.S. Toh ◽  
A.J. Rufaihah ◽  
...  
Keyword(s):  
Cell Death ◽  
Low Temperature ◽  
Survival Rates ◽  
Embryonic Stem ◽  
Slow Cooling ◽  
Trypan Blue Exclusion ◽  
Widespread Application ◽  
Trypan Blue Exclusion Test ◽  
Hes Cells ◽  
Kinetics Of

SummaryA major challenge in the widespread application of hES (human embryonic stem) cells in clinical therapy and basic scientific research is the development of efficient cryopreservation protocols. Conventional slow-cooling protocols utilizing standard cryoprotectant concentrations i.e. 10% (v/v) DMSO, yield extremely low survival rates of less than 5% as reported by previous studies. This study characterized cell death in frozen-thawed hES colonies that were cryopreserved under standard conditions. Surprisingly, our results showed that immediately after post-thaw washing, the overwhelming majority of hES cells were viable (approximately 98%), as assessed by the trypan blue exclusion test. However, when the freshly thawed hES colonies were placed in a 37 °C incubator, there was a gradual reduction in cell viability over time. The kinetics of cell death was drastically slowed down by keeping the freshly thawed hES colonies at 4 °C, with more than 90% of cells remaining viable after 90 min of incubation at 4 °C. This effect was reversible upon re-exposing the cells to physiological temperatures. The vast majority of low temperature-exposed hES colonies gradually underwent cell death upon incubation for a further 90 min at 37 °C. Hence, our observations would strongly suggest involvement of a self-induced apoptotic mechanism, as opposed to cellular necrosis arising from cryoinjury.

scholarly journals Distinct pathways of cholesterol biosynthesis impact on insulin secretion

2014 ◽  
Vol 224 (1) ◽  
pp. 75-84 ◽  
Author(s):  
Juan P Zúñiga-Hertz ◽  
Eduardo Rebelato ◽  
Adam Kassan ◽  
Abdelrahman M Khalifa ◽  
Sameh S Ali ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Insulin Secretion ◽  
Membrane Fluidity ◽  
Structural Integrity ◽  
Cell Function ◽  
Cholesterol Biosynthesis ◽  
Cholesterol Content ◽  
Paramagnetic Resonance ◽  
Cellular Cholesterol ◽  
Plasma Membrane Fluidity

Results from previous investigations have indicated that glucose-stimulated insulin secretion (GSIS) is affected by changes in cholesterol and its intermediates, but the precise link between secretion and cholesterol has not been thoroughly investigated. In this study, we show the contribution of both protein isoprenylation and cholesterol-dependent plasma membrane structural integrity to insulin secretion in INS-1E cells and mouse islets. Acute (2 h) inhibition of hydroxyl-methylglutaryl-CoA reductase by simvastatin (SIM) resulted in inhibition of GSIS without reduction in total cellular cholesterol content. This effect was prevented by cell loading with the isoprenyl molecule geranylgeranyl pyrophosphate. Chronic (24 h) inhibition of cholesterol biosynthesis resulted in inhibition of GSIS with a significant reduction in total cellular cholesterol content, which was also observed after the inhibition of cholesterol biosynthesis downstream of isoprenoid formation. Electron paramagnetic resonance analyses of INS-1E cells showed that the SIM-induced reduction in cholesterol increased plasma membrane fluidity. Thus, the blockade of cholesterol biosynthesis resulted in the reduction of availability of isoprenoids, followed by a reduction in the total cholesterol content associated with an increase in plasma membrane fluidity. Herein, we show the different contributions of cholesterol biosynthesis to GSIS, and propose that isoprenoid molecules and cholesterol-dependent signaling are dual regulators of proper β-cell function.

DMSO Modifies Structural and Functional Properties of RPMI-8402 Cells by Promoting Programmed Cell Death

2003 ◽  
Vol 16 (3) ◽  
pp. 253-259 ◽  
Author(s):  
O. Trubiani ◽  
E. Salvolini ◽  
R. Staffolani ◽  
R. Di Primio ◽  
L. Mazzanti
Keyword(s):  
Nitric Oxide ◽  
Cell Death ◽  
Plasma Membrane ◽  
Programmed Cell Death ◽  
Membrane Fluidity ◽  
Membrane Damage ◽  
Lymphoid Cells ◽  
Plasma Membrane Fluidity ◽  
Structural And Functional Properties ◽  
Intracellular Ca

Apoptosis in lymphoid cells can be induced in different ways depending on cell type and acquired signal. Biochemical modifications occur at an early phase of cell death while at late times the typical morphological features of apoptosis can be visualized. The aim of this study is to verify by multiparametric analyses the plasma membrane fluidity, the intracellular Ca2+ concentration and the nitric oxide synthase (NOS) activity during cell death progression induced by DMSO treatment. The RPMI-8402 human pre-T lymphoblastoid cell line was induced to cell death by DMSO. Analyses rescued at early times of treatment prove a substantial modification of plasma membrane fluidity associated with an increase of intracellular Ca2+. Moreover, these modifications are associated with an up regulation of NOS activity. Our results are consistent with the hypothesis that programmed cell death can be induced by up regulation of the intracellular Ca2+ associated with an increase of cell membrane fluidity. The apoptotic mechanisms seem to involve not only membrane damage and increased intracellular calcium levels but also production of nitric oxide.

vitreal cells and specialized phagocytes in the chick embryo retina: A TEM and SEM study

1990 ◽  
Vol 48 (3) ◽  
pp. 330-331
Author(s):  
M.A. Cuadros ◽  
M.J. Martinez-Guerrero ◽  
A. Rios
Keyword(s):  
Cell Death ◽  
Plasma Membrane ◽  
Acid Phosphatase ◽  
Chick Embryo ◽  
Basement Membrane ◽  
Sem Images ◽  
Intimate Contact ◽  
Cellular Processes ◽  
Sem Study ◽  
Free Cells

In the chick embryo retina (days 3-4 of incubation), coinciding with an increase in cell death, specialized phagocytes characterized by intense acid phosphatase activity have been described. In these preparations, all free cells in the vitreal humor (vitreal cells) were strongly labeled. Conventional TEM and SEM techniques were used to characterize them and attempt to determine their relationship with retinal phagocytes.Two types of vitreal cells were distinguished. The first are located at some distance from the basement membrane of the neuroepithelium, and are rounded, with numerous vacuoles and thin cytoplasmic prolongations. Images of exo- and or endocytosis were frequent; the cells showed a well-developed Golgi apparatus (Fig. 1) In SEM images, the cells was covered with short cellular processes (Fig. 3). Cells lying parallel to or alongside the basement membrane are elongated. The plasma membrane is frequently in intimate contact with the basement membrane. These cells have generally a large cytoplasmic expansion (Fig. 5).

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