Rapid and Automatic Detection of Micronuclei in Binucleated lymphocytes Image

In terms of radiation biological dose estimation, the cytokinesis block micronucleus (CBMN) assay is the internationally recognized dose estimation method. Due to the subjectivity and the time-consuming of manual detection, it cannot meet the needs of rapid standard assay of CBMN. Therefore, in this research work, we combined the convolutional neural network to design a software that can be used for rapid standard automatic detection of micronuclei in Giemsa stained binucleated lymphocytes image. The software analysis workflow is divided into four stages: cell acquisition, adhesive cell masses segmentation, cell type identification, micronucleus counting. After verification, our algorithm can quickly and effectively detect binucleated cells and micronucleus even when the cytoplasm is blurred, multiple micronucleus are attached to each other, or micronucleus is attached to the nucleus. In the test of a large number of random images, the software reached 99.4% of the manual detection in terms of the detection rate of binucleated cell, and the false positive rate of binucleated cell was 14.7%. In terms of micronucleus detection rate, the software reached 115.1% of manual detection, and its false positive rate was 26.2%. The analysis time of each picture is about 0.3s, an order of magnitude faster than conventional method.

Genome instability as an indicator of environmental state in the Murmansk region

The data of various researchers indicate that the combined effects of natural and anthropogenic factors directly affect the nature of the territorial morbidity of the inhabitants of the Far North. In biomonitoring of the environment, a micronucleus test on human cells is widely used to assess the degree of genotoxicity of the comparison areas. The aim of the research is to study the local situation with the accumulation of lesions in the lymphocytes of children living in the Murmansk region using a micronucleus test. As a result of assessing the cytogenetic status of the child population living in the comparison territories (Krasnoshchelye, Lovozero, Umba, Apatity), specific territorial features of the cytogenetic status of children were revealed, which are probably associated with the prevailing morbidity in the territories comparisons. This is probably due to territorial genotoxic and toxic agents. Significant differences in the frequency of occurrence of binucleated lymphocytes with micronuclei in schoolchildren were noted when all cells (mononuclear, binuclear, trinuclear, quadrenuclear cells, as well as cells containing more than 4 nuclei) were taken into account between Krasnoshchelye, Lovozero (U = 45.0, p = 0.0009), Umba (U = 91.0, p = 0.0125) and Apatity (U = 113.0, p = 0.0125). Also, significant differences were noted between the frequency of occurrence of cells with micronuclei among cells that did not respond to the mitogenic signal (mononuclear) and contain more than 2 nuclei in lymphocytes between adolescents from Krasnoshchelye and Apatity (U = 109.0, p = 0.0093) and Umba (U = 73.5, p = 0.0025). When comparing the frequency of occurrence of all types of cells containing micronuclei in the lymphocytes of adolescents, significant differences were revealed between the frequency of occurrence of such cells in adolescents from Apatity and Umba (U = 97.0, p = 0.0036).

Radioprotective effect of pioglitazone against genotoxicity induced by ionizing radiation in human healthy lymphocytes

Objective: Pioglitazone (PG) is used to control high blood sugar in patients with type 2 diabetes mellitus. PG acts as a peroxisome proliferator-activated receptor γ agonist. In addition to the insulin-sensitizing effect, PG possesses anti-inflammatory effect. In this study, the protective effect of PG was evaluated against DNA damage induced by ionizing radiation in human healthy lymphocytes. Methods: The microtubes containing human whole blood were treated with PG at various concentrations (1-50 μM) for three hours. Then, the blood samples were irradiated with X-ray. Lymphocytes were cultured for determining the frequency of micronuclei as a genotoxicity biomarker in binucleated lymphocytes. Results: The mean percentage of micronuclei was significantly increased in human lymphocytes when were exposed to IR, while it was decreased in lymphocytes pre-treated with PG. The maximum reduction in the frequency of micronuclei in irradiated lymphocytes was observed at 5 μM of PG treatment (48% decrease). Conclusion: The anti-inflammatory property is suggested the mechanism action of PG for protection human lymphocytes against genotoxicity induced by ionizing radiation.

Micronucleus as biomarkers of cancer risk in anabolic androgenic steroids users

The use of anabolic androgenic steroids (AAS) has grown among practitioners of recreational bodybuilding, with significant contributions of designer steroids, aiming muscle hypertrophy in healthy subjects. The abusive use of AAS in general is associated with adverse effects; one of the most worrisome is cancer development. The aim of this study was to evaluate the effectiveness of the cytokinesis block micronucleus (CBMN) test in human lymphocytes in identifying risk groups for cancer development in users of AAS. Blood was collected from 15 AAS users bodybuilders (G1), 20 non-users bodybuilders (G2) and 20 non-users sedentary (G3). MN analysis was performed on a minimum of 1000 binucleated lymphocytes. The occurrence of MN was significantly higher ( p < 0.05) in individuals of G1 compared to G2 and G3. The results indicate the sensitivity of CBMN in human lymphocytes in the identification of chromosomal damage in consequence of AAS.

Persistent Polyclonal B Cell Lymphocytosis B Cells Can Be Activated through CD40-CD154 Interaction

Persistent polyclonal B cell lymphocytosis (PPBL) is a rare disorder, diagnosed primarily in adult female smokers and characterized by an expansion of CD19+CD27+IgM+memory B cells, by the presence of binucleated lymphocytes, and by a moderate elevation of serum IgM. The clinical course is usually benign, but it is not known whether or not PPBL might be part of a process leading to the emergence of a malignant proliferative disorder. In this study we sought to investigate the functional response of B cells from patients with PPBL by use of an optimal memory B cell culture model based on the CD40-CD154 interaction. We found that the proliferation of PPBL B cells was almost as important as that of B cells from normal controls, resulting in high immunoglobulin secretion within vitroisotypic switching. We conclude that the CD40-CD154 activation pathway is functional in the memory B cell population of PPBL patients, suggesting that the disorder may be due to either a dysfunction of other cells in the microenvironment or a possible defect in another B cell activation pathway.