Micronucleus Analysis Research Articles

Inheritance patterns of male asexuality in hybrid males of a water frog Pelophylax esculentus.

Gametogenesis produces gametes as a piece of genetic information transmitted to the offspring. While during sexual reproduction, progeny inherits a mix of genetic material from both parents, asexually reproducing organisms transfer a copy of maternal or paternal DNA to the progeny clonally. Parthenogenetic, gynogenetic and hybridogenetic animals have developed various mechanisms of gametogenesis, however, their inheritance is not fully understood. Here, we focused on the inheritance of asexual gametogenesis in hybrid Pelophylax esculentus (RL), emerging after crosses of P. lessonae (LL) and P. ridibundus (RR). To understand the mechanisms of gametogenesis in hybrids, we performed three-generation experiments of sexual P. ridibundus females and hybrids from all-male hybrid populations. Using fluorescent in situ hybridization, micronuclei analysis, flow cytometry and genotyping, we found that most adult hybrid males simultaneously produced two types of clonal sperm. Also, most male tadpole progeny in two successive backcrossed generations simultaneously eliminated L and R parental genomes, while some progeny produced only one type of sperm. We hypothesize that the reproductive variability of males producing two kinds of sperm is an adaptive mechanism to reproduce in mixed populations with P. ridibundus and may explain the extensive distribution of the all-male lineage across the European River Basin.

Cytome analysis (micronuclei and nuclear anomalies) in bioindication of environmental pollution in animals with nuclear erythrocytes

Assessment of cytogenetic homeostasis of indicator animals is of great importance in ecological monitoring. The simplest method of its study is micronucleus analysis. Animals with nuclear erythrocytes are often used as indicator animals. In addition to the micronuclei usually recorded, a wide range of cytological nuclear and cellular abnormalities (cytomic analysis) is encountered when assessing the spontaneous level and under the influence of anthropogenic factors. Spontaneous frequency of cytogenetic disorders in 36 species of fish, amphibians, reptiles and birds was studied. Ecological monitoring of territories of Kazakhstan with different types of pollution (radiation, petrochemical, pesticide, heavy metals, due to rocket and space activities) was carried out with the help of separate species of animals. The results of the study include comparative descriptions, schematics and microphotographs clearly demonstrating a wide range of cytological anomalies of nuclear erythrocytes of animals of different classes. The greatest spectrum of nuclear anomalies in the studied animals was registered at petrochemical and pesticide contamination of territories. Depending on the tasks and climatic-geographical conditions, all investigated species can be used as bioindicators. Testudo horsfieldii is an exception for desert regions due to high spontaneous micronuclei level in this species. A review of the names of the main nuclear anomalies is carried out and variants of its ordering are proposed.

Developmental and heritable genetic defects induced in mice by municipal landfill leachate

Environmental pollution by solid waste leachate is a serious environmental and public health concern. Leachate contamination and pollution of environmental matrices have been reported, but no report of embryotoxic and developmental defects, and heritable transfer of leachate-induced toxicity in mice. We investigated the ability of Aba-Eku landfill leachate to induce embryonic malformations, developmental toxicity, and germline and somatic DNA damage in the F1 of exposed pregnant mice. Pregnant mice (n = 100) were randomly distributed into 5 experimental groups of 20 animals/group and exposed to 0.2 mL of 5–75% concentrations of the leachate (v/v; Aba-Eku landfill leachate: distilled water) by daily gavage from gestational day (GD) zero to postnatal day (PND) 21. A similar treatment was given to pregnant female mice administered with distilled water (negative control). At GD 18, ten dams from the treatment and control groups were sacrificed by cervical dislocation after which the embryos were collected from the uterus for analyses of fetal morphometric and skeletal metamers respectively. We then monitored the developmental conditions of F1 mice from the remaining ten dams until they were weaned at PND 21 and sacrificed at PND 56 and PND 98 for bone marrow micronucleus and spermiogram analyses respectively. We also analyzed the leachate for inorganic and organic pollutants and calculated the Leachate Pollution Index (LPI). The leachate reduced maternal and fetal birth weight and increased fetal mortality and postnatal appearance of physiological markers in the F1 mice. There was a significant increase (p < 0.05) in the frequency of fetal skeletal malformations, micronucleated polychromatic erythrocytes, and apparent decline of epididymal sperm parameters. The concentrations of the inorganic and organic pollutants, and the LPI exceeded standard limits. Exposure of pregnant female mice to Aba-Eku landfill leachate caused embryonic defects and heritable DNA damage in subsequent generations.

Low Dose of Nickel and Benzo [a] Anthracene in Rat-Diet, Induce Apoptosis, Fibrosis, and Initiate Carcinogenesis in Liver via NF-Ƙβ Pathway.

Environmental contaminants such as polycyclic aromatic hydrocarbon (PAH) and heavy metals are major contaminants of food such as fish thus serving as source of exposure to human. This study was designed to evaluate the carcinogenic risk and other risks associated with long-term consumption of environmentally relevant dose of nickel and benzo [a] anthracene in rats. Thirty-six (36) male rats weighing between 80 and 100 g were assigned into 6 groups of 6 animals each; normal, nickel-, and benzo [a] anthracene-exposed groups for 12 and 24 weeks, respectively. Micronucleus and comet analyses were done in the blood, liver, and bone marrow. Liver function, redox, and inflammatory markers (AST, ALT, GGT, SOD, GSH, MDA, protein carbonyl, protein thiol, total protein, IL-10, 1L-1β, TNF-α, TGF-β NF-Ƙβ, and 8-oxodeoxyguansine) were analysed by standard methods. Immuno-histochemical quantification of Bax, Bcl2, and Erk 1/2 as well as mRNA expression of cyclin D1 was done in liver. From the results, weight gain was observed in varying degrees throughout the exposure period. The polychromatic erythrocytes/normochromatic erythrocytes ratio >0.2 indicates no cytotoxic effects on the bone marrow. Percentage-MnPCE in blood significantly (p<0.05) increased throughout exposure duration. Percentage tail DNA in blood was significantly (<0.05) increased at weeks 20 and 24 in the exposed groups and in liver at weeks 12 (16.22±0.47) and 24 (17.00±0.36) of nickel-exposed rats. The aspartate amino transferase (AST):alanine amino transferase (ALT) ratio indicated fatty liver disease in the benzo [a] anthracene (0.90) and acute liver injury in the nickel (>10 times greater than the upper limits of the reference group) exposed groupsduring the first 12 weeks. Observation from the histological and cytological data of the liver revealed the presence of inflammation, fibrosis, and high nuclear/cytoplasmic ratio, respectively, in the nickel and benzo [a] anthracene groups. Only benzo [a] anthracene induced liver oxidative stress with significant (p<0.05) decrease in SOD (0.64±0.02) activity and increase in protein carbonyl (7.60±0.80×10-5) and MDA (57.10±6.64) concentration after 24 weeks. Benzo [a] anthracene up-regulated the cyclin D1 expression and significantly (p<0.05) increased the levels of the cytokines. Nickel and benzo [a] anthracene significantly (p<0.05) increased the Bax (183.45±6.50 and 199.76±10.04) and Erk 1/2 (108.25±6.41 and 136.74±4.22) levels when compared with the control (37.43±22.22 and 60.37±17.86), respectively. Overall result showed that the toxic effects of nickel and benzo [a] anthracene might involve fibrosis, cirrhosis, apoptosis, and inflammation of the liver. As clearly demonstrated in this study, benzo [a] anthracene after the 24 weeks of exposure stimulates carcinogenic process by suppressing the liver antioxidant capacity, altering apoptotic, cell proliferation, and differentiation pathways.

Newly discovered clouting interplay between matrix metalloproteinases structures and novel quaternary Ammonium K21: computational and in-vivo testing

Aims and objectivesTo analyze anti-MMP mode of action of Quaternary Ammonium Silane (QAS, codenamed as k21) by binding onto specific MMP site using computational molecular simulation and Anti-Sortase A (SrtA) mode of action by binding onto specific site using computational molecular simulation.Materials and methodsIn silico Molecular Dynamics (MD) was used to determine the interactions of K21 inside the pocket of the targeted protein (crystal structure of fibroblast collagenase-1 complexed to a diphenyl-ether sulphone based hydroxamic acid; PDB ID: 966C; Crystal structure of MMP-2 active site mutant in complex with APP-derived decapeptide inhibitor. MD simulations were accomplished with the Desmond package in Schrödinger Drug Discovery Suite. Blood samples (~ 0.5 mL) collected into K2EDTA were immediately transferred for further processing using the Litron MicroFlow® PLUS micronucleus analysis kit for mouse blood according to the manufacturer’s instructions. Bacterial Reverse Mutation Test of K21 Molecule was performed to evaluate K21 and any possible metabolites for their potential to induce point mutations in amino acid-requiring strains of Escherichia coli (E. coli) (WP2 uvrA (tryptophan-deficient)).ResultsMolecular Simulation depicted that K21 has a specific pocket binding on various MMPs and SrtA surfaces producing a classical clouting effect. K21 did not induce micronuclei, which are the result of chromosomal damage or damage to the mitotic apparatus, in the peripheral blood reticulocytes of male and female CD-1 mice when administered by oral gavage up to the maximum recommended dose of 2000 mg/kg. The test item, K21, was not mutagenic to Salmonella typhimurium (S. typhimurium) strains TA98, TA100, TA1535 and TA1537 and E. coli strain WP2 uvrA in the absence and presence of metabolic activation when tested up to the limit of cytotoxicity or solubility under the conditions of the test.ConclusionK21 could serve as a potent protease inhibitor maintaining the physical and biochemical properties of dental structures.

Risk Characterization of Botanical Extracts Containing Hydroxyanthracenes as Determined by a Validated Micronucleus In Vitro Assay

Extracts of Rheum palmatum L., Rhamnus purshiana DC., Rhamnus frangula L., and Cassia senna L. are used in traditional medicine thanks to their beneficial properties. These species contain hydroxyanthracene derivatives, considered genotoxic and possibly related to colorectal cancer development. This research aimed to study, using a micronucleus assay in vitro, the genotoxic potential of Rheum palmatum L., Rhamnus purshiana DC., Rhamnus frangula L. (bark), and Cassia senna L. (leaves and fruits) extracts. The extracts were evaluated at different concentrations: from 0 to 2000 µg/mL for Rhamnus purshiana DC, from 0 to 2500 µg/mL for Rheum palmatum L. and Rhamnus frangula L., and from 0 to 5000 µg/mL for Cassia senna L. The cytokinesis-block proliferation index was calculated to analyse if the used concentrations showed cytotoxicity. The hydroxyanthracene content varied between 0.06% and 0.23% for aloe-emodin, and between 0.07% and 0.16% for emodin and rhein. No cytotoxic effect was detected at any of these concentrations. Micronucleus analyses showed a lack of genotoxicity for all the extracts tested. These results show that Rheum palmatum L., Rhamnus purshiana DC, Rhamnus frangula L., and Cassia senna L. extracts do not induce genotoxicity since no increase in micronuclei formation in human lymphocytes in vitro was detected.

Efficiency of toxicological tests on Rural Workers exposed to testicides

The objective of this study was to compare the results of two methods to verify acute pesticide poisoning, the analysis of micronucleus (MN) of cells in the oral mucosa and the measurement of Plasma Cholinesterase (BuChE) and Erythrocyte Cholinesterase (AChE) in rural workers exposed to pesticides. As methodological procedures, two groups with 27 individuals each were determined. Blood samples were collected to measure AChE and BuChE, Alkaline Phosphatase, Gamma Glutamil Transferase (Gamma GT), Oxaloacetic Transaminase (TGO) and Pyruvic Transaminase (TGP), for the exposed group, at two moments: after the moment of application of the insecticide-type pesticide and after the application of any other type of pesticide. In the unexposed group, only one collection was carried out. For MN analysis, cells were collected from the buccal mucosa from the inner region of the cheek. The group exposed to pesticides had a higher number of MN when compared to the group not exposed to pesticides. There was no significant difference in AChE between the exposed group and the unexposed group at the first moment (p = 0.847) and similarly for BuChE (p = 0.070).

Visualization of Fagopyrum esculentum and Fagopyrum tataricum Chromosomes and Micronuclei.

This chapter presents the squash chromosome preparation technique for Fagopyrum esculentum and F. tataricum, using the root tips as the source of the material. Using an optimized version of this method, the chromosomes are free of cytoplasmic debris and are spread evenly on the glass slide. What comes of it is the possibility to make observations of the chromosome number and structure at the metaphase stage. This technique's modified version allows micronuclei analysis in interphase cells of buckwheats.

Ameliorative effects of Tinospora cordifolia against cytogenotoxicity induced by lead acetate in Mus musculus

Studies concerning exposure of humans to lead (Pb) during daily activities are sometimes leading to many complications. Pb is a poisonous heavy metal; its toxicity can change the antioxidant balance in biological systems. T. cordifolia is rich in phenolic acids and flavonoids which exhibit a wide range of biological effects and act as a natural antioxidant. The objective of the research work- first is to investigate the genotoxic potential of Pb-acetate on Bone marrow cells (BMCs), Second is to investigate the antigenotoxic potential of T. cordifolia against genotoxic induced by Pb-acetate in BMCs of Mus musculus (Swiss albino mice). For the research experiment, Adult Mus musculus of the same age group were selected andwere distributed into four groups (Group I, II, III &amp;IV). After the completion of dose treatment, the experimental animals were euthanized by cervical dislocation and exposed both femora for the analysis of Micronucleus (Mn) in Polychromatic and Normochromatic erythrocytes. For the assessment of frequency of Mn in polychromatic or normochromatic erythrocytes 800 - 1000 cells were screened randomly under the compound microscope at 40X. and screening was done by image J microscopy software. Results of the experiment showed that the total frequency of Mn in polychromatic and normochromatic erythrocytes was 0.27 ± 0.07 in Group I, 0.18 ± 0.06 in Group II, 4.32 ± 0.32in Group III, and 0.94 ± 0.15 in Group IV and it is a significant difference to control and treated group at p &lt; 0.05 or &lt; 0.01. Hence, the results of the research experiment indicate that T. cordifolia stem aqueous extract has the potency to ameliorate and minimize the genotoxic induced by Pb-acetate in M. musculus (Swiss albino mice).

Оцінка генетичної мінливості племінних стад амурського сазана (Cyprinus rubrofuscus Lacépède, 1803)

Purpose. To study the genetic peculiarities of pedigree tocksof Amur carp and evaluate its genetic variability by analyzing distribution of alleles and genotypes by the specific protein systems and cytogenetic parameters. Methodology. Analysis of polymorphism of protein systems was performed using electrophoresis in polyacrylamide gel. As protein markers to evaluate the genetic structure of Amur carp stocks, the separation of allelic and genotypic frequencies by loci, which encode a number of fish blood proteins, were used: transferrin (TF), albumin (ALB) and esterase (EST, 3.1.1.1). Cytogenetic analysis was performed using the micronucleus test and analysis of apoptosisfrequencies. Collection and processing of biological material of fish as well as statistical processing of the obtained data was performed using generally accepted methods. Findings. A comprehensive analysis of the genetic structure of Amur carp from three farms in Ukraine was carried out by protein systems and cytogenetic markers. The results of the study showed that all pedigree groups of Amur carp were characterized by a deviation towards the predominance of heterozygotes (Fis = from –0.084 to –0.344). A minimal imbalance was observed in the group of carp from the farm “Karpatskyi Vodogray” LLC (Fis = –0.084). The maximum violation of the genetic balance was found in carp from the farm of JSC “Sumyrybhosp” (Fis = –0.308) and in the group from the experimental farm “Veliky Lyubin” (Fis = –0.344). In general, the analysis of the protein systems of fish blood showed a high level of heterogeneity of pedigree stocks of Amur carp from different farms in Ukraine. The analysis of frequencies of cytogenetic parameters of Amur carp from three fish farms showed that fish from fish farms of JSC “Sumyrybhosp” and PFE “Dzherelo” were characterized by a lower frequency of erythrocytes with micronuclei (EMN) (3.3±0.3 ‰), (3.2±0.3 ‰), lymphocytes with micronuclei (LMN) (2.1±0, 2 ‰), (1.9±0.2 ‰), and apoptosis (4.2±0.3‰), (4,3±0,3‰) compared to the group from fish farm “Karpatskyi Vodogray” LLC, where these values were as follows: EMN (4.7±0.3 ‰), LMN (2.4±0.2‰), apoptosis (5.6±0.4‰). These results indicates a lower level of destabilization of chromosomal apparatus of Amur carp from the fish farm JSC “Sumyrybhosp” at the time of the study. Originality. A comprehensive evaluation of the level of variability of the genetic structure of pedigree stocks of Amur carp from different regions of Ukraine was performed for the first time. Practical Value. Pedigree socksof Amur carp have important practical value in selective breeding work with carps, including works for obtaining hybrid lines to increase the resistance of breeding material. Keywords: Amur carp, genetic structure, locus, alleles, genotype, heterozygosity, micronucleus test, cytogenetic analysis.

Abstract 310: CHAF1A promotes the translesion DNA synthesis pathway in response to DNA replication stress

Abstract Objective: Translesion DNA synthesis (TLS) pathway allows cancer cells to bypass the damage, allowing the blocked replication fork to move forward and helping cancer cells avoid genomic instability and cell death caused. Although some PCNA partners have previously been reported to participate in the TLS pathway, the mechanisms that regulate the TLS pathway remain unclear. Here, we revealed that CHAF1A is a new important regulator of TLS pathway. Methods: Cell models of CHAF1A gene knockdown (A549, KYSE510, KYSE450, etc.) were constructed by siRNA/shRNA knockdown method. We used DNA damage stimulation (HU, APH and UV) combined with western blotting to explore the effect of CHAF1A on PCNA monoubiquitination. The interaction of CHAF1A and its partners was demonstrated by immunoprecipitation and Duolink PLA assay. Cell fractionation assay was employed to measure protein binding to chromatin. DNA fiber assay was used to observe the condition of DNA replication fork. Micronucleus analysis was used to quantify cell micronucleus. The sensitivity of cells to DNA replication stress was tested by cloning formation assay and cell death assay. Kaplan-Meier survival analysis were used to analyze the clinical significance of CHAF1A expression. Results: CHAF1A enhances the interaction between PCNA and E3 ubiquitin protein ligase RAD18 and promotes PCNA monoubiquitination, thereby promoting the recruitment of Y-family DNA polymerase Pol η and enhancing cancer cell resistance to stimuli that trigger replication fork blockade. Mechanistically, CHAF1A-mediated PCNA monoubiquitination is independent of CHAF1A-PCNA interaction. CHAF1A interacts with both RAD18 and replication protein A2 (RPA2), mediating RAD18 binding on chromatin in response to DNA replication stress. Moreover, high expression of CHAF1A is significantly associated with poor prognosis in cancer patients. Conclusion: CHAF1A is a key TLS pathway regulator that is essential for cancer cell survival under DNA replication stress. The overall survival time of patients with high CHAF1A expression in lung, liver and esophageal cancers was significantly reduced. CHAF1A can be used as a key potential anti-tumor target, which has important clinical significance for enhancing the sensitivity of patients to radiotherapy and chemotherapy (This work was supported by the National Natural Science Foundation of China (82273108 and 82173034), the Innovative Team Grant of Guangdong Department of Education (2021KCXTD005), the Science and Technology Special Fund project of Guangdong Province in 2021 (No. 210729156901797) and the 2020 Li Ka Shing Foundation Cross-Disciplinary Research Grant (2020LKSFG07B)). Citation Format: Bing Wen, Li-Yan Xu, En-Min Li. CHAF1A promotes the translesion DNA synthesis pathway in response to DNA replication stress [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 310.

Implementation of effect biomarkers in human biomonitoring studies: A systematic approach synergizing toxicological and epidemiological knowledge.

Human biomonitoring (HBM) studies have highlighted widespread daily exposure to environmental chemicals. Some of these are suspected to contribute to adverse health outcomes such as reproductive, neurological, and metabolic disorders, among other developmental and chronic impairments. One of the objectives of the H2020 European Human Biomonitoring Initiative (HBM4EU) was the development of informative effect biomarkers for application in a more systematic and harmonized way in large-scale European HBM studies. The inclusion of effect biomarkers would complement exposure data with mechanistically-based information on early and late adverse effects. For this purpose, a stepwise strategy was developed to identify and implement a panel of validated effect biomarkers in European HBM studies. This work offers an overview of the complete procedure followed, from comprehensive literature search strategies, selection of criteria for effect biomarkers and their classification and prioritization, based on toxicological data and adverse outcomes, to pilot studies for their analytical, physiological, and epidemiological validation. We present the example of one study that demonstrated the mediating role of the effect biomarker status of brain-derived neurotrophic factor BDNF in the longitudinal association between infant bisphenol A (BPA) exposure and behavioral function in adolescence. A panel of effect biomarkers has been implemented in the HBM4EU Aligned Studies as main outcomes, including traditional oxidative stress, reproductive, and thyroid hormone biomarkers. Novel biomarkers of effect, such as DNA methylation status of BDNF and kisspeptin (KISS) genes were also evaluated as molecular markers of neurological and reproductive health, respectively. A panel of effect biomarkers has also been applied in HBM4EU occupational studies, such as micronucleus analysis in lymphocytes and reticulocytes, whole blood comet assay, and malondialdehyde, 8-oxo-2'-deoxyguanosine and untargeted metabolomic profile in urine, to investigate, for example, biological changes in response to hexavalent chromium Cr(VI) exposure. The use of effect biomarkers in HBM4EU has demonstrated their ability to detect early biological effects of chemical exposure and to identify subgroups that are at higher risk. The roadmap developed in HBM4EU confirms the utility of effect biomarkers, and support one of the main objectives of HBM research, which is to link exposure biomarkers to mechanistically validated effect and susceptibility biomarkers in order to better understand the public health implications of human exposure to environmental chemicals.

Fabrication of hydroxyapatite-based nano-gold and nano-silver-doped bioceramic bone grafts: Enhanced mechanostructure, cell viability, and nuclear abnormality properties.

In this study, nano-gold (nAu) and nano-silver (nAg) were doped at the molar ratios of Molar5-Molar30 to the Hydroxyapatite (HAp)-based bioceramic bone graft synthesized by the sol-gel method. The effects of nAu and nAg on structural, mechanical, cell viability, and nuclear abnormality of the synthesized bioceramic grafts were evaluated. The chemical and morphological properties of the bone grafts after production were examined through XRD and SEM-EDX analyses and mechanical tests. To determine the biocompatibility of the bone grafts, cell viability tests were performed using human fibroblast cells. In the cytotoxicity analyses, only HAp and HAp-nAu5 grafts did not show toxicological properties at any concentration, while HAp-nAg5 among the nAg-containing grafts gave the best results at the 200-100 μg/mL concentrations and showed significant cytotoxicity in human fibroblast cells. The other nAu-containing grafts showed toxicological properties in the concentration range of 200-50 μg/mL and nAg-containing grafts in the concentration range of 200-100 μg/mL against the negative control. The micronucleus (MN) analyses showed that the lowest total MN and L (lobbed) amounts, while the lowest total N (notched) amount, was obtained from the only HAp graft. It was found that the nAg-doped bone grafts gave higher total MN, L, and N amounts compared to the nAu-doped bone grafts. Furthermore, while the mean nuclear abnormality (NA) values of all grafts gave close results, the highest values were again obtained from the nAg-doped bone grafts.

Possibilities of using micronucleus analysis to detect gene mutations in animals

Relevance. The main purpose of micronuclear analysis is to identify substances that cause cytotoxic damage and lead to the formation of micronuclei containing lagging fragments of chromosomes or whole chromosomes. For the study of chromosomal disorders, micronuclear analysis is now very often used, since it is quite easy to perform and does not require large costs.Methods and results. Micronuclei are cytoplasmic chromatin-containing bodies formed when delayed in anaphase or telophase during cell division or fragmentation of the nucleus during apoptosis. The method of micronuclear analysis is used as a biomarker of chromosomal aberrations in the study of mutagenicity, as well as a marker of the development of oncological diseases. With quantitative changes in DNA in the cell, the formation of micronuclei occurs. It is their presence that is an indicator that the body was exposed to environmental pollution (pesticides, pesticides, radiation, heavy metals, etc.), leading to inflammatory and pathological processes. Recently, many studies have been conducted aimed at studying the influence of environmental factors on the genetic apparatus of the body. Under the influence of mutagens, micronuclei are formed in proliferating cells, the counting of which allows for the diagnosis of both genotoxic stress and genetic instability of the organism. To determine early genetic changes in humans and animals, sensitive and labor-intensive methods are needed, which can be attributed to micronuclear analysis. Recently, micronuclear analysis has become relevant for its use in the livestock sector, mainly in research aimed at quality control of livestock products used in the future for human nutrition.

Biodose Tools: an R shiny application for biological dosimetry

Introduction In the event of a radiological accident or incident, the aim of biological dosimetry is to convert the yield of a specific biomarker of exposure to ionizing radiation into an absorbed dose. Since the 1980s, various tools have been used to deal with the statistical procedures needed for biological dosimetry, and in general those who made several calculations for different biomarkers were based on closed source software. Here we present a new open source program, Biodose Tools, that has been developed under the umbrella of RENEB (Running the European Network of Biological and retrospective Physical dosimetry). Materials and methods The application has been developed using the R programming language and the shiny package as a framework to create a user-friendly online solution. Since no unique method exists for the different mathematical processes, several meetings and periodic correspondence were held in order to reach a consensus on the solutions to be implemented. Results The current version 3.6.1 supports dose-effect fitting for dicentric and translocation assay. For dose estimation Biodose Tools implements those methods indicated in international guidelines and a specific method to assess heterogeneous exposures. The app can include information on the irradiation conditions to generate the calibration curve. Also, in the dose estimate, information about the accident can be included as well as the explanation of the results obtained. Because the app allows generating a report in various formats, it allows traceability of each biological dosimetry study carried out. The app has been used globally in different exercises and training, which has made it possible to find errors and improve the app itself. There are some features that still need consensus, such as curve fitting and dose estimation using micronucleus analysis. It is also planned to include a package dedicated to interlaboratory comparisons and the incorporation of Bayesian methods for dose estimation. Conclusion Biodose Tools provides an open-source solution for biological dosimetry laboratories. The consensus reached helps to harmonize the way in which uncertainties are calculated. In addition, because each laboratory can download and customize the app’s source code, it offers a platform to integrate new features.

Micronuclei analysis in mice peripheral blood exposed to polarized polychromatic noncoherent light (Bioptron® Light)

The increase in cancer in recent years suggests an inadvertent exposure to agents that cause genetic damage. The polarized polychromatic noncoherent light Bioptron® lamp is used to accelerate healing, among other therapeutic applications and its potential carcinogenic effects as a mitogenic agent have not been explored. The objective was to evaluate the genotoxicity of the Bioptron light therapy by means of the micronucleus assay in mouse erythrocytes. Male SKH1 hairless mice were randomly divided into six groups (5 mice/group); Group 1: negative control received ambient light; Group 2: positive control was exposed to ultraviolet light lamp A (UV-A) for 80 min; Experimental Groups 3–6 were exposed to the Bioptron lamp light for 10, 20, 40 and 80 min, respectively. Exposures in all groups were once a day for 4 days and blood smears were performed daily for 5 days and subsequently read with a microscope equipped with epifluorescence. The values of micronucleated erythrocytes (MNE), micronucleated polychromatic erythrocytes (MNPCE) and the proportion of polychromatic erythrocytes (PCE) were determined. The study group that received the UV-A light was the only one that increased MNE and MNPCE values, while in the groups exposed to the Bioptron lamp and the negative control did not show increases in any of the sampling days. In conclusion, under the conditions presented here, our results suggest that the light of the Bioptron lamp does not cause damage to the genetic material of SKH1 mice, by means of the micronucleus test in peripheral blood.

Micronucleus test using formalin-fixed rat glandular stomach and colon

BackgroundGenotoxicity in tissues other than hematopoietic tissues, such as the liver and gastrointestinal (GI) tract, is an important focus in the risk assessment of chemicals in humans. We previously developed a rat micronucleus test for the GI tract, which is the first contact tissue where chemicals are introduced into the body through oral exposure. Target cells were obtained from fresh tissue samples by ethylenediaminetetraacetic acid disodium salt (EDTA) treatment. As an improvement to this method, we have used formalin-fixed tissues instead of fresh tissues; this approach can be used for tissues that are sampled from other toxicological tests and that are archived for several years. This new method can be used for examining micronucleus induction retrospectively when needed. In the present study, we compared the performance of the EDTA method and the new method with formalin-fixed tissues (formalin-fixation method).ResultsHistological examination showed that both the EDTA and formalin-fixation methods could be used for collecting cells located in or above the proliferative zone of the GI tract tissues of rats. In addition, the collected cells were similar in shape. We conducted micronucleus tests with rat GI tract tissues by the two methods using model chemicals, which were used as positive control chemicals (a combination of diethylnitrosamine, 1,2-dimethylhydrazine dihydrochloride, and potassium bromate). The two methods showed similar results. We additionally evaluated the aging effect of tissues stored in formalin fixative. The results showed that 1 year of storage did not affect the frequency of micronucleated cells.ConclusionThe equivalence of the EDTA and formalin-fixation methods was confirmed, and micronucleus analysis was possible up to at least 1 year after formalin fixation of the GI tract, indicating that the formalin-fixation method is valuable for the rat GI tract micronucleus test.

Micronucleus test and assessment of changes in peripheral blood erythrocytes of dominant animal species in the Turkestan region

The article presents the results of a study of peripheral blood erythrocytes of dominant animal species in the Turkestan region. The relevance of this work is due to the construction of a nuclear power plant on the territory of Uzbekistan (Jizzakh region, Lake Tuzkan), since the areas of the Turkestan region fall under the potential influence of nuclear power plants and biota objects require monitoring at the ‘zero stage’ before the construction of a nuclear power plant. The data obtained are of great importance in assessing the impact of technogenic factors on biota. Given the location of uranium deposits in the Turkestan region, the uranium content was determined in samples (bones, soft tissues) of amphibians and fish. It has been established that the concentration of uranium in samples of biological objects is within the normal range. We have carried out work on the analysis of erythrocytes and micronuclei in the following biota representatives: desert lacerta (lat. Eremias intermedia), yellow ground squirrel (lat. Spermophilus or Citellus), lake frog (lat. Pelophylax ridibundus), rudd (lat. Redd pisces), snakehead (lat. Channa argus). According to the results of studies in the desert lacerta (lat. Eremias intermedia) and lake frog (lat. Pelophylax ridibundus), various pathological conditions of erythrocytes and micronuclei were observed in 11% of the total number of blood products studied.

GENETIC CHARACTERISTICS OF PADDLEFISH (POLYODON SPATHULA (WALBAUM, 1792)) BROOD STOCKS

GENETIC CHARACTERISTICS OF PADDLEFISH (POLYODON SPATHULA (WALBAUM, 1792)) BROOD STOCKS

Study of physiological and cytogenetic changes to males patients with diabetic mellitus type2 by used the body mass index

The study included 20 patients and have ranged in age from (30-75 years), compared with a control groups which included 12 individual, with ages have ranged from (38-70) years. The study investigate biochemical parameters included measuring the levels of each of (Total Cholesterol , Triglyceride , low-density lipoproteins, high density lipoproteins , lipoproteins very low density, and measuring hormone levels Testosterone, and estimate the damage in DNA and measuring the Micronucleus and Mitotic index and analysis of Comet Assay in lymphocytes for patients and control: The results of our study showed a high change (P&lt; 0.05) in the level of each of (cholesterol, triglycerides , low-density lipoproteins , lipoproteins very low density , and the low significant (P&lt; 0.05) level of high density lipoproteins and of the Testosterone hormone .The results showed a high significant (P&lt; 0.05) in the Mitotic index and Micronucleus , appearance of numbers low in the number the Nuclear division index, where results showed a high significant (P&lt; 0.05) in damage to the DNA molecule by comet assay when compared with control group.