scholarly journals Analysis of radiation-induced apoptosis in human lymphocytes: Flow cytometry using Annexin V and propidium iodide versus the neutral comet assay

Cytometry ◽  
2002 ◽  
Vol 48 (1) ◽  
pp. 14-19 ◽  
Author(s):  
R.C. Wilkins ◽  
B.C. Kutzner ◽  
M. Truong ◽  
J. Sanchez-Dardon ◽  
J.R.N. McLean
Keyword(s):  
Flow Cytometry ◽  
Comet Assay ◽  
Propidium Iodide ◽  
Human Lymphocytes ◽  
Annexin V ◽  
Neutral Comet Assay ◽  
Radiation Induced ◽  
Induced Apoptosis

Radiation-Induced Apoptosis Measured in TK6 Human B Lymphoblast Cells Using the Comet Assay

1993 ◽  
Vol 136 (1) ◽  
pp. 130 ◽  
Author(s):  
Peggy L. Olive ◽  
Garnet Frazer ◽  
Judit P. Banáth ◽  
Judit P. Banath
Keyword(s):  
Comet Assay ◽  
Radiation Induced ◽  
Induced Apoptosis ◽  
Lymphoblast Cells

Modification of radiation-induced apoptosis in radiation- or hyperthermia-adapted human lymphocytes

1994 ◽  
Vol 72 (11-12) ◽  
pp. 475-482 ◽  
Author(s):  
S. P. Cregan ◽  
D. R. Boreham ◽  
P. R. Walker ◽  
D. L. Brown ◽  
R. E. J. Mitchel
Keyword(s):  
Dna Damage ◽  
Ionizing Radiation ◽  
Adaptive Response ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Fluorescence Analysis ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Protective Mechanism ◽  
Radiation Induced ◽  
Induced Apoptosis

We have investigated the influence of the cellular adaptive response to ionizing radiation on radiation-induced apoptosis in human cells. The adaptive response is believed to be a protective mechanism that confers resistance to the detrimental effects of ionizing radiation and that can be induced by different agents, including hyperthermia and radiation. We have used fluorescence analysis of DNA unwinding (FADU) to assay the induction of apoptosis in human peripheral blood lymphocytes by ionizing radiation. Using the FADU assay, we have observed the initial radiation-induced DNA damage, its subsequent disappearance due to enzymatic repair, and its time- and dose-dependent reappearance. We believe this reappearance of DNA damage to be indicative of the DNA fragmentation event associated with apoptosis. This interpretation has been supported at the individual cell level using an in situ terminal deoxynucleotidyl transferase (TDT) assay (Apoptag, Oncor Inc.), which detects the 3′-hydroxyl ends of fragmented DNA, and by fluorescence analysis of nuclear morphology in Hoechst 33258 stained cells. Pretreatment of cells with low-dose γ-radiation (0.1 Gy) or mild hyperthermia (40 °C for 30 min) altered the extent of radiation-induced (3 Gy) apoptosis. Both pretreatments sensitized lymphocytes to become apoptotic after the 3-Gy radiation exposure. This sensitization may represent an adaptive response mechanism that reduces the risk that genetically damaged cells will proliferate. The ability to modify the probability of radiation-induced apoptosis may lower the cancer risk from a radiation exposure.Key words: apoptosis, adaptive response, ionizing radiation, hyperthermia.

Dose – response for radiation-induced apoptosis, residual 53BP1 foci and DNA-loop relaxation in human lymphocytes

2005 ◽  
Vol 81 (2) ◽  
pp. 125-138 ◽  
Author(s):  
J Torudd ◽  
M Protopopova ◽  
R Sarimov ◽  
J Nygren ◽  
S Eriksson ◽  
...  
Keyword(s):  
Dose Response ◽  
Human Lymphocytes ◽  
Dna Loop ◽  
Radiation Induced ◽  
Induced Apoptosis

Angelica Polysaccharide and TPO Have a Protective Effect On Hcmv-Induced Apoptosis In Megakaryocytes

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3553-3553
Author(s):  
Mo Yang ◽  
Jian Liang Chen ◽  
Jie yu Ye ◽  
Su yi Li ◽  
En yu Liang ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Protective Effect ◽  
Caspase 3 ◽  
Hcmv Infection ◽  
Annexin V ◽  
Angelica Sinensis ◽  
Control Group ◽  
Mock Control ◽  
Apoptotic Effect ◽  
Induced Apoptosis

Abstract Human cytomegalovirus (hCMV) infection is often associated with thrombocytopenia. Megakaryocytes may be one of the major sites of hCMV infection, then inducing this cell apoptosis. Angelica Sinensis (Danggui) is an important ingredient of many commonly used herbal Medicine for promoting blood production. Our previous study has showed that the hematopoietic effect of Angelica Sinensis is related to its constituent, angelica polysaccharide (APS) (Yang M et al, J Ethnopharma, 2009). This present study investigated the anti-apoptotic effect of APS and TPO on hCMV-induced apoptosis in megakaryocytes. Human bone marrow mononuclear cells (MNC) or megakaryocytic cell line CHRF-288-11 and hCMV AD169 strain were co-cultured in this study. hCMV significantly inhibited the formation of CFU-MK as shown in three different concentrations of viral infection groups (103, 104 and 105 pfu/ml), compared with blank control and mock control (n=10, P<0.05). hCMV also significantly inhibited the growth of CHRF cells in these three different concentrations after incubation for 3 days, which compared with control group (n=10, P<0.01). hCMV DNA and mRNA were also positively detected in CHRF cells and the cells of CFU-MK with IS-PCR and RT-PCR respectively, while it was negative in blank and mock control groups. We further studied the effect of APS and TPO on CFU-MK formation. Results showed that APS (50 ug/ml) like TPO (50 ng/ml) enhanced hCMV-reduced CFU-MK (P=0.05, n=6). CHRF cells were also analyzed by Annexin V/PI with flow cytometry at day 3 after infection with hCMV AD169. The percentage of apoptotic cells in group of 103 pfu/ml was 19.0 ± 2.0%; The group of 104 pfu/ml was 23.0 ± 1.5%; The group of 105 pfu/ml was 28.0 ± 3.0%. The control group was 2.0 ± 0.5%. The apoptotic cells were confirmed by morphologic observation. In addition, apoptotic signals from megakaryocytic surface, cytoplasma and mitochondria were detected in hCMV infected cells by flow cytometry with Caspase-3 and JC-1 assay. Compared to mock infection control at day 5, Annexin-V positive cells population increased by 58%; active caspase-3 signal increased by 120% in viable cell population; and cell population with damaged mitochondial membrane showed a 5-times increase. Moreover, the anti-apoptotic effect of APS and TPO on CHRF cells was also demonstrated by using Annexin-V assay. Our studies showed that hCMV induces the apoptosis in megakaryocytes via mitochondrial and caspase-3 signaling, and angelica polysaccharide (APS) like TPO has a protective effect on hCMV-induced apoptosis in these cells. Disclosures: No relevant conflicts of interest to declare.

Radiation-Induced Apoptosis in Human Lymphocytes and Lymphoma Cells Critically Relies on the Up-Regulation of CD95/Fas/APO-1 Ligand

1998 ◽  
Vol 149 (6) ◽  
pp. 588 ◽  
Author(s):  
C. Belka ◽  
P. Marini ◽  
W. Budach ◽  
K. Schulze-Osthoff ◽  
F. Lang ◽  
...  
Keyword(s):  
Human Lymphocytes ◽  
Lymphoma Cells ◽  
Radiation Induced ◽  
Induced Apoptosis

scholarly journals Flow cytometry evaluation of HeLa S3 cell death induced by gamma-radiation

2004 ◽  
Vol 23 (2) ◽  
pp. 135-142 ◽  
Author(s):  
Ana Niciforovic ◽  
Bozidarka Zaric ◽  
Aleksandra Dakic ◽  
Nevena Tisma ◽  
Marija Radojcic
Keyword(s):  
Flow Cytometry ◽  
Cell Death ◽  
Radiation Dose ◽  
Gamma Radiation ◽  
Fluorescent Microscopy ◽  
Annexin V ◽  
Hela S3 ◽  
Induced Apoptosis ◽  
60Co Gamma Radiation

In this study we followed the effects of radiation on human uterin cervix HeLa S3 cells viability, morphology and DNA structure 2-96 hours after treatment with 2-10 Gy from 60Co gamma radiation source. Staining of cells with Annexin V-FITC and propidium iodide showed very low degree of radiation-induced apoptosis. The prevailing form of HeLa S3 cell death according to flow-cytometry, DNA fragmentation and fluorescent microscopy, was necrosis. The gamma-radiation dose necessary to induce 50% of necrosis (termed DD50) was twice higher compared to dose that induced 50% inhibition of cell proliferation (LD50). These in vitro data suggested, that the increase in radiation dose might eradicate tumor cells, rather than just control their proliferation and growth.

scholarly journals Evaluation of Immobilization of Selected Peat-Isolated Yeast Strains of the Species Candida albicans and Candida subhashii on the Surface of Artificial Support Materials Used for Biotrickling Filtration

Processes ◽  
2020 ◽  
Vol 8 (7) ◽  
pp. 801
Author(s):  
Milena Marycz ◽  
Anna Brillowska-Dąbrowska ◽  
Jacek Gębicki
Keyword(s):  
Flow Cytometry ◽  
Candida Albicans ◽  
Propidium Iodide ◽  
Annexin V ◽  
Support Materials ◽  
Hydrophobic Compounds ◽  
Yeast Strains ◽  
Different Types ◽  
Materials Used ◽  
Hydrophilic And Hydrophobic Compounds

The paper describes the process of n-butanol abatement by unicellular fungi, able to deplete n-butanol content in gas, by using n-butanol as source of carbon. Isolated and identified fungi species Candida albicans and Candida subhashii were subjected to a viability process via assimilation of carbon from hydrophilic and hydrophobic compounds. The isolates, which exhibited the ability to assimilate carbon, were immobilized on four different types of artificial support materials used for biotrickling filtration. Application of optical microscopy, flow cytometry and the tests employing propidium iodide and annexin V revealed viability of the fungi isolated on support materials’ surfaces at the average level of 95%. The proposed method of immobilization and its evaluation appeared to be effective, cheap and fast. Based on performed comparative analyses, it was shown that polyurethane foam and Bialecki rings (25 × 25) could be attractive support materials in biotrickling filtration.

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