scholarly journals Short Assay Design for Micronucleus Detection in Human Lymphocytes

2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Guido Rincón ◽  
Claudia Sánchez
Keyword(s):  
Incubation Period ◽  
Human Lymphocytes ◽  
Micronucleus Assay ◽  
High Sensitivity ◽  
Standard Protocol ◽  
Review Of Literature ◽  
Genotoxic Effects ◽  
Cell Incubation ◽  
Assay Time ◽  
Assay Design

There has been a constant need to develop new and faster cytogenetic assays to measure the instability induced by genotoxic agents in the field of cytogenetic research, an example of which is the micronucleus assay. Micronuclei are fragments or complete chromosomes that remain in the cytoplasm during mitosis. With their high sensitivity and specificity detection, their presence can indicate environmental and occupational genotoxic effects. However, the prolonged periods of cell incubation this assay necessitates are costly and extensive. Hence, it is essential to develop an improved assay that can achieve standardization by being reproducible in practice. The standard protocol for the detection of micronuclei in lymphocytes uses a total assay time of 72 hours. Theoretically, it is possible to reduce the incubation period, and consequently, the total assay time, considering a lymphocyte, completes its mitosis in 24 hours. This study, after careful review of literature, proposes an experimental design to reduce the incubation period and demonstrates its usefulness in practice through the design of a collaborative trial.

scholarly journals Genotoxic evaluation of newly synthesized organometallic compounds of tin

2015 ◽  
Vol 17 (3) ◽  
pp. 574-582
Keyword(s):  
Cytotoxic Effect ◽  
Human Lymphocytes ◽  
Micronucleus Assay ◽  
Organometallic Compounds ◽  
Proliferation Index ◽  
Tripodal Ligands ◽  
Genotoxic Effects

<p>The genotoxic effects of organometallic tin(II) and tin(IV) complexes namely L<sub>OEt</sub>SnCl (<strong>5</strong>), L*<sub>OEt</sub>SnCl (<strong>6</strong>), L<sub>OEt</sub>SnPh<sub>3</sub> (<strong>7</strong>), L*<sub>OEt</sub>SnPh<sub>3</sub> (<strong>8</strong>), incorporating the oxygen tripodal ligands [(<em>η</em><sup>5</sup>-C<sub>5</sub>R<sub>5</sub>)Co{P(OEt)<sub>2</sub>O}<sub>3</sub>]<sup>-</sup>, {R = H, (L<sub>OEt</sub><sup>-</sup>) (<strong>3</strong>); R = Me (L*<sub>OEt</sub><sup>-</sup>) (<strong>4</strong>)} (Klaui type ligands), were investigated using the Cytokinesis Block Micronucleus assay in human lymphocytes cultures. For comparison the precursors NaL<sub>OEt</sub> (<strong>3</strong>), NaL*<sub>OEt</sub> (<strong>4</strong>), SnCl<sub>2</sub>&bull;2H<sub>2</sub>O (<strong>1</strong>) and Ph<sub>3</sub>SnCl (<strong>2</strong>), were also studied.</p> <p>Statistically significant differences in comparison with the control in the micronuclei frequencies were seen at the concentrations: 75 μM for complex (<strong>5</strong>), 50 μM for complex (<strong>6</strong>), 20, 50, 75 μM for complex (<strong>8</strong>). No statistically significant differences were observed between controls and all the rest tested concentrations for all chemicals examined.</p> <div> <p>The cytotoxic effect was evaluated by the Cytokinesis Block Proliferation Index. Regarding this index, the precursor (<strong>1</strong>) is not cytotoxic at all tested concentrations. Complexes (<strong>3</strong>), (<strong>4</strong>) and (<strong>5</strong>) induced cytotoxicity at the concentrations of 20, 50 and 75 μM, while complexes (<strong>6</strong>) and (<strong>8</strong>) were cytotoxic at all tested concentrations. Complex (<strong>7</strong>) was cytotoxic at 5, 10 and 20 μM but extremely toxic at 50 and 75 μM. Finally complex (<strong>2</strong>) was extremely toxic at all tested concentrations except at 1 μM.&nbsp;</p> </div> <p>&nbsp;</p>

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.

scholarly journals A Tool Derived from the Vicia faba Micronucleus Assay, to Assess Genotoxicity, Cytotoxicity or Biostimulation of Novel Compounds Used in Agriculture

Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 321
Author(s):  
Perrine Klein ◽  
Lorelei Chauvey ◽  
Jean Kallerhoff ◽  
Eric Pinelli ◽  
Marie Morard ◽  
...  
Keyword(s):  
Vicia Faba ◽  
Maleic Hydrazide ◽  
Lead Nitrate ◽  
Micronucleus Assay ◽  
Growth Potential ◽  
Protective Effects ◽  
Genotoxic Effects ◽  
Cytotoxic Effects ◽  
Conventional Agriculture ◽  
Rapid Tests

The increased use of biostimulants in conventional agriculture and organic farming requires the implementation of rapid tests to determine their effectiveness in enhancing plant growth and protection against abiotic stresses. However, their innocuity to plant health has rarely been demonstrated. We used the Vicia faba Micronucleus Assay, as described by the standard AFNOR EN ISO 29200(2020-05) to reveal biostimulant, genotoxic and cytotoxic effects of four commercialized wood-based products by comparing mitotic indices and micronucleus frequencies with respect to the controls. Neither genotoxicity, as measured by micronucleus frequency (MN), nor cytotoxicity, assessed by Mitotic index counts, was observed. Additionally, one of these stimulants (BHS®) conferred protective effects against contaminants (maleic hydrazide or lead nitrate). We describe that plotting micronuclei frequency against mitotic indices allows discrimination between cytotoxic/genotoxic effects from growth levels. Vicia faba experiments were successfully transposed to other agronomical important crops such as corn and sunflower. This technique can be valuable to industrials, to assess growth, potential cytoxicity and genotoxicity effects of any new biostimulant or organic.

scholarly journals Micronucleus assay in human lymphocytes after exposure to alloxydim sodium herbicide in vitro

2014 ◽  
Vol 67 (6) ◽  
pp. 1059-1066 ◽  
Author(s):  
Dilek Akyıl ◽  
Arzu Özkara ◽  
S. Feyza Erdoğmuş ◽  
Yasin Eren ◽  
Muhsin Konuk ◽  
...  
Keyword(s):  
Human Lymphocytes ◽  
Micronucleus Assay

Diagnosis and t-staging of gastric cancer: comparison of standard computed tomography and computed-tomographic pneumogastrography

2021 ◽  
Vol 23 (2) ◽  
pp. 99-106
Author(s):  
Inna D. Amelina ◽  
Lev N. Shevkunov ◽  
Aleksey M. Karachun ◽  
Alexander E. Mikhnin ◽  
Denis V. Nesterov
Keyword(s):  
Gastric Cancer ◽  
Computed Tomography ◽  
Three Dimensional ◽  
High Sensitivity ◽  
Subtotal Resection ◽  
Standard Protocol ◽  
Computed Tomographic ◽  
T Stage ◽  
Significant Difference ◽  
Virtual Gastroscopy

The advantages of computed tomographic pneumogastrography with the possibility of three-dimensional visualization and virtual gastroscopy in diagnosing gastric cancer are considered. The study included 479 patients with histologically diagnosed gastric cancer who were treated at the National Medical Research Center of Oncology named after N.N. Petrov from 2011 to 2018. 232 patients received preoperative chemotherapy. All patients underwent surgery: 70 in the volume of endoscopic dissection, 40 proximal subtotal resection, 166 distal subtotal resection, 203 gastrectomy. All patients at the preoperative stage underwent staging computed tomography on a 64-slice X-ray computed tomograph: 208 patients underwent computed tomography according to the standard protocol without targeted preparation of the stomach for the study, 271 patients with targeted preparation of the stomach for the study according to the computed tomographic pneumogastrography protocol. The sensitivity of the computed tomography in assessing the T-stage was assessed by comparison with pathomorphological data. Of the 208 patients who underwent computed tomography according to the standard protocol, a gastric cancer was detected in 111 (53.4%), out of 271 patients who underwent computed tomography pneumogastrography, a gastric cancer was detected in 267 (98.52%), which is a statistically significant difference in comparing computed tomography methods (Pearson, 144.223, df = 1; p 0.001). There are statistically significant differences when comparing computed tomography according to the standard protocol and computed tomographic pneumogastrography in detecting gastric cancer for all tumor categories: T/yT1 8.2 and 94.4% (Pearson, 99.205, df = 1; p 0.001), T/yT2 47.8 and 100% (Pearson, 24.681, df = 1; p 0.001), T/yT3 72.3 and 100% (Pearson, 33.114, df = 1; p 0.001), T/yT4 90.0 and 100% (Pearson, 4.789, df = 1; p = 0.029) respectively. There are also statistically significant differences when comparing the sensitivity of computed tomography according to the standard protocol and computed tomographic pneumogastrography in determining tumor invasion for all tumor categories: T/yT1 0 and 69.4% (Pearson, 67.880, df = 1; p 0.001), T/yT2 26.1 and 71.1% (Pearson, 11.666, df = 1; p 0.001), T/yT3 32.9 and 84.6% (Pearson, 54.900, df = 1; p 0.001), T/yT4 73.3 and 95.7% (Pearson, 7.916, df = 1; p = 0.005) respectively. In general, the sensitivity of the computed tomography according to the standard protocol for determining the T-stage of gastric cancer was 28.4%, computed tomographic pneumogastrography 77.1% (Pearson, 113.505, df = 1; p 0.001). Computed tomographic pneumogastrography with the possibility of three-dimensional visualization and virtual gastroscopy significantly increases the indicators of the effectiveness of diagnosing gastric cancer both early forms (category T1) and with deeper invasion (categories T2T4), demonstrates high sensitivity in determining T/yT-stages.

Genotoxic Effects of Amplitude-Modulated Microwaves on Human Lymphocytes Exposed in Vitro under Controlled Conditions

1995 ◽  
Vol 14 (3) ◽  
pp. 157-164 ◽  
Author(s):  
G. D'ambrosio ◽  
M. B. Lioi ◽  
R. Massa ◽  
M. R. Scarfi ◽  
O. Zeni
Keyword(s):  
Human Lymphocytes ◽  
Genotoxic Effects ◽  
Controlled Conditions

scholarly journals Reduction of doxorubicin-induced genotoxicity by Handroanthus impetiginosus in mouse bone marrow revealed by micronucleus assay

2017 ◽  
Vol 78 (1) ◽  
pp. 1-12
Author(s):  
M. F. G. Boriollo ◽  
T. A. Silva ◽  
M. F. Rodrigues-Netto ◽  
J. J. Silva ◽  
M. B. Marques ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Micronucleus Assay ◽  
Chemical Compounds ◽  
Chemotherapeutic Agents ◽  
Mouse Bone Marrow ◽  
Genotoxic Effects ◽  
Dose Time ◽  
Polychromatic Erythrocytes ◽  
And Gender ◽  
Handroanthus Impetiginosus

Abstract Handroanthus impetiginosus has long been used in traditional medicine and various studies have determined the presence of bioactive chemical compounds and potential phytotherapeutics. In this study, the genotoxicity of the lyophilized tincture of H. impetiginosus bark (THI) was evaluated in mouse bone marrow using micronucleus assays. The interaction between THI and genotoxic effects induced by the chemotherapeutic agent, doxorubicin (DXR), was also analyzed. Experimental groups were evaluated 24 to 48 h after treatment with N-nitroso-N-ethylurea (NEU; 50 mg/kg), DXR (5 mg/kg), sodium chloride (NaCl; 150 mM), and THI (0.5-2 g/kg). Antigenotoxic assays were carried out using THI (0.5 g/kg) in combination with NEU or DXR. Analysis of the micronucleated polychromatic erythrocytes (MNPCEs) indicated no significant differences between treatment doses of THI (0.5-2 g/kg) and NaCl. Polychromatic erythrocyte (PCE) to normochromatic erythrocyte (NCE) ratios did not indicate any statistical differences between DXR and THI or NaCl, but there were differences between THI and NaCl. A significant reduction in MNPCEs and PCE/NCE ratios was observed when THI was administered in combination with DXR. This study suggested the absence of THI genotoxicity that was dose-, time-, and gender-independent and the presence of moderate systemic toxicity that was dose-independent, but time- and gender-dependent. The combination of THI and DXR also suggested antigenotoxic effects, indicating that THI reduced genotoxic effects induced by chemotherapeutic agents.

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