A Cytogenetic Approach to the Study of Genotoxic Effects of Fungicides: An in Vitro Study in Lymphocyte Cultures with Thiophanate-methyl

1996 ◽  
Vol 24 (4) ◽  
pp. 597-601
Author(s):  
Patrizia Hrelia ◽  
Carmela Fimognari ◽  
Fernanda Vigagni ◽  
Francesca Maffei ◽  
Giorgio Cantelli-Forti
Keyword(s):  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
In Situ Hybridisation ◽  
In Vitro Study ◽  
Peripheral Blood Lymphocytes ◽  
Fluorescence In Situ Hybridisation ◽  
Thiophanate Methyl ◽  
Mechanisms Of Toxicity ◽  
Genetic Level

This study was designed to evaluate the mechanisms of genetic damage by fungicides in cultured human peripheral blood lymphocytes by means of a molecular cytogenetic approach. For example, thiophanate-methyl (30μg/ml-300μg/ml) was shown to significantly induce chromosome aberrations and micronuclei in human lymphocytes cultured in vitro. Fluorescence in situ hybridisation with centromeric DNA probes demonstrated that most micronuclei induced by thiophanate-methyl did not show any centromeric signals, indicating a relatively stronger clastogenic activity. Results obtained with thiophanate-methyl showed that a comprehensive examination of the mechanisms of toxicity at the genetic level provides valuable information, which is of importance in the safety assessment of the fungicide.

scholarly journals In vitro effect of tuibur (tobacco brew) on the viability of human blood lymphocytes

2017 ◽  
Vol 17 (1) ◽  
pp. 19-24 ◽  
Author(s):  
B. Lalruatfela ◽  
Jennifer Zoremsiami ◽  
Ganesh Chandra Jagetia
Keyword(s):  
Adverse Effect ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
In Vitro Study ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Human Peripheral Blood Lymphocytes ◽  
Long Time ◽  
Vitro Effect

The use of tobacco and its products are known to cause many illnesses including cancer. A smokeless tobacco locally manufactured called tuibur (tobacco brew) has been consumed by the Mizos from a very long time. In this experiment we aim to determine the cytotoxicity of tuibur by an in vitro study on tuibur-treated human peripheral blood lymphocytes. We have found that 24 h treatment of human lymphocytes with two grades of commercial tuibur and nicotine showed a concentration dependent decrease in cell viability. We, therefore, concluded that as the in vitro use of tuibur has an adverse effect on cell survival, its consumption might have potential side effects on the health of the users.

Soluble CD14 Protects Human Lymphocytes from Apoptosis

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2566-2566
Author(s):  
Elizabeth Naparstek ◽  
Benjamin Sredni ◽  
Eti Zigman ◽  
Gali Senyor ◽  
Boris Tartakovsky
Keyword(s):  
Human Peripheral Blood ◽  
Protective Efficacy ◽  
Human Lymphocytes ◽  
Membrane Integrity ◽  
Peripheral Blood Lymphocytes ◽  
Apoptotic Cells ◽  
Soluble Cd14 ◽  
Apoptotic Pathway ◽  
Cell Membrane Integrity

Abstract CD14, a 56 Kd glycoprotein, typically present on myeloid cells, has been traditionally associated with innate immunity and pattern recognition. Recently its membrane bound form has been shown to be involved in apoptosis, as a tethering receptor for apoptotic cells on the surface of phagocytes-in this case with the purpose of removing apoptotic cells, and also as a surface molecule involved in protection from apoptosis of monocytes, neutrophils and recently on enterocytes, challenged with LPS. Our aim was to evaluate the possible involvement of the soluble CD14 in the apoptotic pathway of human lymphocytes. Methods: Freshly obtained human peripheral blood lymphocytes were cultured in vitro with gliotoxin, an apoptotic inducer. Human recombinant CD14 was added to the culture at physiological concentrations (10μg/ml-0.5 μg/ml) and apoptosis was assessed by cell membrane integrity using 7AAD, mitochondrial membrane potential by DiOC6(3) and cytoplasm shrinkage by cell size scatter analysis. Results: Using DiOC6(3) we were able to show that human lymphocytes cultured in the presence of gliotoxin contained 63.8%±21 apoptotic cells, as opposed to 12.2%±11.5 in control cultures. Addition of recombinant human CD14 at a concentration of 10 mg/ml neutralized the apoptotic effect of gliotoxin back to 20.2%±10 (p<0.003). This inhibitory effect was blocked by CD14-specific monoclonal antibodies, but not by control antibodies. We then identified and synthesized the fragment within the CD14 molecule that was responsible for this apoptosis protective effect, and demonstrated its comparable protective efficacy in vitro as shown in figure 1. The figure clearly reveals that this specific peptide, as opposed to the scrambled peptide, protected the lymphocytes form apoptosis, similarly to the full CD14 protein. Same results were obtained using 7AAD and cytoplasm shrinkage. Conclusion: Our data thus suggest that circulating CD14 may play an important role in the prevention of apoptosis of lymphocytes and perhaps of other cells. Figure Figure

scholarly journals Cytotoxic, clastogenic and genotoxic effects of cis-tetraammine(oxalato)ruthenium(III) dithionate on human peripheral blood lymphocytes

2020 ◽  
Vol 42 ◽  
pp. e50517
Author(s):  
Manuela da Rocha Matos Rezende ◽  
Vivianne de Souza Velozo-Sá ◽  
Cesar Augusto Sam Tiago Vilanova-Costa ◽  
Elisangela Silveira-Lacerda
Keyword(s):  
Comet Assay ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Low Frequency ◽  
Peripheral Blood Lymphocytes ◽  
Negative Control ◽  
Blood Lymphocytes ◽  
Human Peripheral Blood Lymphocytes

There is a concern about stablishing the clinical risk of drugs used for cancer treatment. In this study, the cytotoxic, clastogenic and genotoxic properties of cis-tetraammine(oxalato)ruthenium(III) dithionite - cis-[Ru(C2O4)(NH3)4]2(S2O6), were evaluated in vitro in human lymphocytes. The mitotic index (MI), chromosomal aberrations (CA) and DNA damage by comet assay were also analyzed. The MTT test revealed that the ruthenium compound showed a slight cytotoxic effect at the highest concentration tested. The IC50 value for the compound after 24 hours of exposure was 185.4 µM. The MI values of human peripheral blood lymphocytes treated with 0.015, 0.15, 1.5 and 150 µM of cis-[Ru(C2O4)(NH3)4]2(S2O6) were 6.1, 3.9, 3.2 and 0.2%, respectively. The lowest concentration, 0.015 µM, did not show any cytotoxic activity. The CA values for the 0.015, 0.15 and 1.5 µM concentrations presented low frequency (1.5, 1.6 and 2.3%, respectively), and did not express clastogenic activity when compared to the negative control, although it was observed clastogenic activity in the highest concentration tested (150 µM). The results obtained by the comet assay suggest that this compound does not present genotoxic activity at lower concentrations. The results show that cis-[Ru(C2O4)(NH3)4]2(S2O6) has no cytotoxic, clastogenic or genotoxic in vitro effects at concentrations less than or equal to 0.015 µM. This information proves as promising in the treatment of cancer and is crucial for future trials.

scholarly journals Cycling Characteristics of Human Lymphocytes In Vitro

Blood ◽  
1974 ◽  
Vol 44 (1) ◽  
pp. 99-107 ◽  
Author(s):  
Lewis M. Schiffer ◽  
Arnold M. Markoe ◽  
Alan Winkelstein ◽  
Janet S. R. Nelson ◽  
John M. Mikulla
Keyword(s):  
Cell Cycle ◽  
Dna Polymerase ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Peripheral Blood Lymphocytes ◽  
Cell Cycle Time ◽  
Daughter Cells ◽  
Human Peripheral Blood Lymphocytes ◽  
A New Technique

Abstract The cycling characteristics of human peripheral blood lymphocytes in phytohemagglutinin-stimulated cultures were examined by means of a new technique employing radioautographic methods which assay the fraction of cells whose nuclei contain DNA polymerase utilizing exogenous and/or endogenous DNA primer-template capability. The fraction of cells labeled by these techniques increases 5-11 hr prior to the onset of DNA synthesis. Estimates of cell cycle time, G1 time, and fraction of cycling cells are made for the first 4 days in culture. Evidence is presented to support the hypothesis that daughter cells of the first divisions may be retired from cycle by virtue of loss of primer-template availability, rather than by loss of DNA polymerase.

scholarly journals In-VitroCarbofuran Induced Genotoxicity in Human Lymphocytes and Its Mitigation by Vitamins C and E

2012 ◽  
Vol 32 (3) ◽  
pp. 153-163 ◽  
Author(s):  
Ratnesh Kumar Sharma ◽  
Bechan Sharma
Keyword(s):  
Dna Damage ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Natural Antioxidants ◽  
Underlying Mechanism ◽  
Positive Control ◽  
Peripheral Blood Lymphocytes ◽  
Dependent Manner ◽  
Concentration Dependent Manner

Various efforts have been made in past in order to predict the underlying mechanism of pesticide-induced toxicity usingin vitroand animal models, however, these predictions may or may not be directly correlated with humans. The present study was designed to investigate the carbofuran induced genotoxicity and its amelioration by vitamins C and E by treating human peripheral blood lymphocytes (PBLs) with different concentrations (0, 0.5, 1.25, 2.5, 3.75 and 5.0 μM) of this compound. The treatment of PBLs with carbofuran displayed significant DNA damage in concentration dependent manner. The carbofuran induced genotoxicity could be ameliorated to considerable extent by pretreatment of PBLs with equimolar (10 μM) concentration of each of the vitamins C and E; the magnitude of protection by vitamin E being higher than by vitamin C. Also, it was found that the level of protection by these vitamins was higher when PBLs were treated with lower concentrations of pesticide. The significant DNA damage as observed by H2O2, a positive control in the present study, and its amelioration by natural antioxidants (vitamins C and E) lend an evidence to suggest that carbofuran would have caused genotoxicity via pesticide induced oxidative stress.

scholarly journals Micronucleus Assay as a Biological Indicator for In Vitro Exposure of Human Lymphocytes to Gamma Rays

2012 ◽  
Vol 6 (1) ◽  
pp. 51-55
Author(s):  
Abbas N. Balasem
Keyword(s):  
Gamma Rays ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Micronucleus Assay ◽  
Biological Indicator ◽  
Peripheral Blood Lymphocytes ◽  
Radiation Accidents ◽  
In Vitro Exposure ◽  
Different Doses

Micronucleus Assay was employed to detect the effects of acute exposure of human peripheral blood lymphocytes in vitro to Cs -137 gamma rays. Human whole blood samples were irradiated with different doses of gamma rays namely ) 0.1, 0.2, 0.3, 0.4, 0.5, 1.00) Gy, respectively in addition to a control non-irradiated sample. The samples were tissue cultured and cytokinesis blocked method was used to investigate the frequency of micronuclei. In vitro exposure of lymphocytes to this doses led to elevation of micronuclei in comparison with non –irradiated samples However, inclusion of mono-, tri-,and quadrinucleated cells in micronucleus assay probably gives more satisfying result than restriction the test on binucleated cells. Computed programmed were employed to establish dose – response relationships to be used as biological dosimeter during radiation accidents.

scholarly journals Estimating genotoxic effects of anticancer drugs using cytokinesis-block micronucleus assay on human lymphocytes

2019 ◽  
Vol 4 (3) ◽  
pp. 95-101
Author(s):  
V. I. Minina ◽  
V. Yu. Buslaev
Keyword(s):  
Anticancer Drugs ◽  
Human Lymphocytes ◽  
In Situ Hybridisation ◽  
Micronucleus Assay ◽  
Cochrane Library ◽  
Dependent Manner ◽  
Fluorescence In Situ Hybridisation ◽  
Genotoxic Effects ◽  
Cbmn Assay

Here we review the current experience of using cytokinesis-block micronucleus (CBMN) assay on cultures of human lymphocytes to evaluate genotoxic effects of anticancer drugs. Having performed search in PubMed, Scopus, Web of Science, TOXLINE, and the Cochrane Library, we identified a total of 172 relevant studies. Out of them, 89 were conducted in vitro, and 41 were published within the last decade. The mentioned studies concordantly demonstrated a significant increase in micronuclei, protrusions, nucleoplasmic bridges, and a decrease in proliferation in cells treated with anticancer drugs in a time- and dose-dependent manner. Notably, the results of CBMN assay are consistent with the data obtained from other cytogenetic techniques (comet assay, chromosomal aberration analysis, analysis of mutations in housekeeping genes, and fluorescence in situ hybridisation). Conclusion. CBMN assay permits a reliable evaluation of the mutagenic effects related to anticancer drugs.

Cytotoxic Effects of Trimethyltin Chloride on Human Peripheral Blood Lymphocytes in vitro

1989 ◽  
Vol 8 (5) ◽  
pp. 349-353 ◽  
Author(s):  
B.B. Ghosh ◽  
G. Talukder ◽  
A. Sharma
Keyword(s):  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Peripheral Blood Lymphocytes ◽  
Sister Chromatid Exchanges ◽  
Blood Lymphocytes ◽  
Human Peripheral Blood Lymphocytes ◽  
Abnormal Cells ◽  
Trimethyltin Chloride

Trimethyltin chloride was found to induce cytotoxic damage in vitro in human peripheral blood lymphocytes. Two concentrations (0.5 μg and 1.0 μg) were added to lymphocytes from male and female subjects in mitogen stimulated and serum supplemented culture medium for 72 h. A dose-related increase of inhibition of replication index (P < 0.01) and cell division (P < 0.001) was observed. The frequencies of abnormal cells and chromosomal aberrations such as chromatid and chromosome breaks, dicentrics, triradial and quadriradial configurations were increased significantly (P < 0.001), as were micronucleus counts (P < 0.001) and sister chromatid exchanges (P < 0.001). Endoreduplication, an extremely rare spontaneous event in human lymphocytes, was observed in a few cases at all dose levels. The effects were more pronounced in lymphocytes obtained from habitual smokers.

Chlorobenzenes, lindane and dieldrin induce apoptotic alterations in human peripheral blood lymphocytes (in vitro study)

2013 ◽  
Vol 36 (3) ◽  
pp. 979-988 ◽  
Author(s):  
Jaromir Michałowicz ◽  
Katarzyna Mokra ◽  
Karolina Rosiak ◽  
Paulina Sicińska ◽  
Bożena Bukowska
Keyword(s):  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
In Vitro Study ◽  
Peripheral Blood Lymphocytes ◽  
Vitro Study ◽  
Blood Lymphocytes ◽  
Human Peripheral Blood Lymphocytes
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