Frequency Of Micronuclei Research Articles

Toxicity assessment of a phytomedicine based on an almond extract from the fruit of Balanites aegyptiaca (L.) DELILE (Zygophyllaceae) intended for helminthiases care in Burkina Faso

Balanites aegyptiaca (L.) Delile (Zygophyllaceae) is a medicinal plant used in traditional medicine in Burkina Faso to treat helminthiasis. Using an ethnopharmacological approach, the "Institut de Recherche en Sciences de la Santé (IRSS)" has developed a phytomedicine based on an almond extract from the fruit of this plant for the treatment of helminthiasis. This study aimed to assess the acute and subacute oral toxicity and in vivo mutagenicity of phytomedicine. The acute toxicity study was carried out following OECD guideline 423 by administering a single dose of 2000 mg/kg body weight (bw) of phytomedicine orally to female mice. For subacute toxicity, OECD guideline 407 was used. Four batches of 10 rats (5 males and 5 females) were used, including a control batch and three treated batches receiving a daily oral administration of the phytomedicine at doses of 250, 500, or 1000 mg/kg bw for 28 days. The mutagenicity test was performed according to OECD guideline 474; five batches of 10 mice (5 males and 5 females) were given oral doses of 500, 1000 or 2000 mg/kg bw of phytomedicine, colchicine as a positive control at 5mg/kg bw and distilled water as a negative control. In the acute toxicity test, the LD50 was estimated at 5000 mg/kg bw. The subacute toxicity study showed no mortality or signs of toxicity in rats. Biochemical analysis revealed no significant differences between control rats and those treated with the phytomedicine for glucose, creatinine, total cholesterol, triglycerides, AST, chlorine, calcium, and potassium. On the other hand, serum levels of ALT, total protein, PO43-, and Na+ were significantly reduced in some treated rats. The mutagenicity test showed no change in micronucleus frequency between treated and control mice. This study showed that phytomedicine based on an extract of B. aegyptiaca fines would present less danger to users. Keywords: Balanites aegyptiaca, acute toxicity, subacute toxicity, micronucleated erythrocytes, phytomedicine.

ANTIGENOTOXIC ACTIVITY OF GRACILARIA SP. ON ERYTHROCYTES OF NILE TILAPIA EXPOSED BY METHOMYL-BASED PESTICIDE

This research investigates the biochemical profile, antioxidant properties, and antigenotoxic activity of red seaweed (Gracilaria sp.) extract on micronuclei formation in peripheral blood erythrocytes of tilapia (Oreochromis niloticus) exposed to a methomyl-based pesticide. Results revealed a diverse phytochemical profile in the red seaweed extract, with the presence of alkaloids, triterpenoids, steroids, tannins, coumarins, terpenoids, quinine, phytosteroids, flavonoids, and phlobtannins. GC-MS analysis identified compounds such as Heptadecane, Neophytadine, Hexadecanoic acid, Oleic acid, and Adipic acid in the sample. The antioxidant activity exhibited a concentration-dependent response, demonstrating increased strength with higher extract concentrations. Antigenotoxic analysis indicated a significant reduction in micronuclei frequency in peripheral erythrocytes of tilapia. In conclusion, the research underscores the potential of red seaweed extract as a valuable natural resource with antioxidant properties and a mitigating effect on genotoxicity induced by pesticide exposure in aquatic organisms.

Rosa damascena Mill. Essential Oil: Analysis of In Vitro and In Vivo Genotoxic and Cytotoxic Potentials by Employing Three Cytogenetic Endpoints.

The highly valued oil of Rosa damascena Mill. (Rosaceae), widely used in high perfumery, cosmetics, and other spheres of human life, obliges us to know and study the safety profile of the product obtained from the water-steam distillation of fresh rose petals. The genotoxicity of the essential oil (EsO) has not been thoroughly studied despite its wide range of applications. That predetermined the object of this study-to evaluate, through classical cytogenetic methods, the possible cytotoxic/genotoxic activities of R. damascena Mill. EsO (EsORdm) in three different test systems: plant root meristem cells, mammalian bone marrow cells, and human lymphocyte cultures. The rose essential oil showed varying concentration- and time-dependent cytotoxic and genotoxic effects depending on the test system used, and it was established that the oil showed moderate cytotoxicity in lymphocyte cultures and non-high cytotoxicity in ICR mice but none in barley. Both barley and human lymphocytes showed a genotoxic effect with a dose-dependent increase in chromosomal aberrations (CAs) and a substantial rise in micronucleus (MN) frequency, while no genotoxicity was observed in bone marrow cells at the applied concentrations. Human lymphocytes exhibited the highest susceptibility to cytotoxic and genotoxic actions of the EsO. As a valuable plant-derived aromatic product with versatile uses in human life, R. damascena Mill. essential oil should be used in an appropriate concentration range tailored to cellular sensitivity.

Genetic instability of a single exposure to sevoflurane at different concentrations in monitored mice.

Sevoflurane is an inhalation anesthetic widely used for general anesthesia, but its genotoxic potential is controversial in clinical studies. It is unknown whether the effects are due to surgery or the anesthetic. Thus, for the first time, the present study investigated genotoxicity in peripheral blood cells and in target organs (liver, lung, and kidney) and micronucleus (MN) in the bone marrow of a single exposure to sevoflurane at three different concentrations in monitored mice. Ninety Swiss mice were distributed into the following groups: exposure to sevoflurane at 3.3% (low), 4.5% (intermediate), and 6.0% (high) in 40% oxygen (O2) for 2 h; negative control (no exposure); negative control with O2; and positive control. The exposed animals were heated, monitored for vital signs (temperature, O2 saturation, heart rate/pulse, and respiratory rate), and anesthetized via a modern low-flow digital system. Mice were euthanized 2 and 24 h after exposure for evaluation by the comet assay and MN test, respectively. No DNA damage occurred in the 3.3% group for any of the organs evaluated, and no genotoxic or mutagenic effects were observed at any sevoflurane concentration in the peripheral blood or liver cells. However, a significant increase in DNA damage was observed at higher concentrations in kidney (4.5%) and lung cells (6.0%) and in the MN frequency (groups 4.5% and 6.0%). No cytotoxicity or histological alterations were observed. In conclusion, high concentrations of sevoflurane induce DNA damage, but concentrations equivalent to those used in clinical practice do not demonstrate genotoxic or mutagenic effects.

Studies on ameliorative potentials of quercetin nanoparticles against imidacloprid induced subacute genotoxicity and histopathological alteration in Swiss albino mice

ABSTRACT Objective Genotoxicity assays include micronucleus test, comet assay, and malformed sperm head used to investigate the protective potential of quercetin and quercetin nanoparticles against imidacloprid-induced genotoxicity in Swiss albino mice. Method The ionic gelation procedure was used to synthesize the quercetin nanoparticles and characterized for their hydrodynamic diameter, zeta potential, SEM, TEM, FT-IR, and encapsulation efficiency. Total 48 mice were taken in eight groups with six animals in each group. Group 1, 2, 3 and 4 received 3% gum acacia, 22 mg/kg imidacloprid (IMI), 25 mg/kg quercetin (Que) and 25 mg/kg quercetin nanoparticles high dose [QNPs (HD)], respectively. Group 5, 6, 7 and 8 received 22 mg/kg imidacloprid + 25 mg/kg quercetin (IMI + Que), 22 mg/kg imidaclopid + 25 mg/kg quercetin nanoparticles [IMI + QNPs (HD)], 22 mg/kg imidacloprid + 12.5 mg/kg quercetin nanoparticle medium dose [IMI + QNPs (MD)] and 22 mg/kg imidacloprid + 6.25 mg/kg quercetin nanoparticles low dose [IMI + QNPs (LD)], respectively. Result The imidacloprid causes genotoxicity in bone marrow cells by increasing the frequency of micronuclei and the comet tail length. Additionally, imidacloprid is mutagenic to germ cells, as determined by a test for aberrant sperm heads. Both quercetin and quercetin nanoparticles lessen the genotoxicity that imidacloprid induces when administered together or separately. Histopathological findings also revealed degenerative changes in bone marrow and testes in imidacloprid administered group as compared to control. Conclusion Quercetin and quercetin nanoparticles showed marked ameliorative effect by restoring the degenerative changes produced by imidacloprid.

Morphological and cellular effects in Boana faber tadpoles (Anura: Hylidae) exposed to atrazine-based herbicide and glyphosate-based herbicide and their mixtures.

Atrazine and glyphosate are considered some of the main pollutants for aquatic ecosystems, directly and indirectly affecting non-target organisms, such as amphibians. This study aimed to evaluate the sublethal effects of different concentrations of atrazine-based herbicide (ABH) and glyphosate-based herbicide (GBH) commercial formulations, both individually and in a mixture, through toxicity tests on the larval stage of Boana faber. Tadpoles were exposed to concentrations of ABH (2, 9.33, 10.40, 47.21, and 240μg L-1) and GBH (65, 144, 280, 500, and 1000μg L-1), as well as a mixture ABH + GBH, for 7days. Although survival and swimming activity were not significantly affected by herbicide exposure, tadpoles in all treatments showed damage to the mouth and intestine, changes in size and mass, and an increase in the frequency of micronuclei and other nuclear abnormalities. Despite differences in some variables analyzed, it is not possible to definitively state that there is a difference in the toxicity of these two herbicides, as both caused morphological damage and were cyto-genotoxic. Our findings suggest that exposure to commercial formulations of these herbicides, whether alone or in mixture, can directly impact the quality of life of B. faber tadpoles.

Study Using the Micronuclear Test of Radiosensitivity and Induction Radiation of Adaptive Response in Peripheral Blood Lymphocytes of Patients with Oncological Diseases

Radiosensitivity to low and medium doses of X-ray radiation (XR) and the ability to induce a radiation adaptive response (RAR) of lymphocytes during in vitro irradiation of peripheral blood of patients with cancer were studied. The criterion for cytogenetic damage was the frequency of micronuclei (MN) in cytochalasin-blocked binucleate lymphocytes in culture. It was found that the spontaneous level of cytogenetic damage in the lymphocytes of patients was 2.6 times higher than in healthy volunteers, and there was also significant interindividual variability in values compared to the control cohort. There were no differences in mean values for radiosensitivity to low and medium doses of XR between the study groups. There was no correlation between the spontaneous level of MN in lymphocytes and the radiosensitivity of individuals in both groups. RAR was induced with the same frequency and to the same extent in lymphocytes from both patients and healthy individuals.

The essential role of TTC28 in maintaining chromosomal stability via HSPA8 chaperone-mediated autophagy

There are three distinct forms of autophagy, namely, macroautophagy, microautophagy, and HSPA8 chaperone-mediated autophagy (CMA). While macroautophagy is widely recognized as a regulator of chromosomal instability (CIN) through various pathways, the contributions of CMA and microautophagy to CIN remain uncertain. TTC28, a conserved gene in vertebrates, is frequently mutated and down-regulated in numerous human cancers. This study presents findings demonstrating the interaction between human tetratricopeptide repeat domain 28 (TTC28) and heat shock protein member 8 (HSPA8) and lysosomal-associated membrane protein 2A proteins. The tetratricopeptide repeat domains of TTC28 bind to the C-terminal motif (PTIEEVD) in HSPA8, resulting in the subsequent degradation of TTC28 via CMA/microautophagy. Notably, the baseline frequency of micronuclei (FMN) in human cancer cells with TTC28 knockout cells was three times greater than that in cells with wild-type TTC28 (7.7% vs. 2.3%, P = 4.86E-09). Furthermore, the overexpression of Ttc28 mitigated the impact of TTC28 knockout on FMN (11.9% vs. 4.8%, P = 2.83E-11). Our findings also demonstrate that CMA has a protective effect on genome stability and that TTC28 plays an essential role in the effect of CMA. These results were further supported by the quantification of γH2AX and comet analyses and the analysis of The Cancer Genome Atlas data via bioinformatics. Mechanistically, TTC28 regulates mitosis and cytokinesis, which are involved in the maintenance of genome integrity by CMA. In conclusion, our study demonstrated that TTC28 is not only an HSPA8-mediated CMA/microautophagy substrate but also essential for maintaining chromosomal stability via CMA. Comprehensive TTC28 downregulation may lead to CIN in cancer cells.

Non-Destructive Biomarkers in Non-Target Species Earthworm Lumbricus terrestris for Assessment of Different Agrochemicals

In many agroecosystems, agrochemicals are widely used to control crop pests, but often affect many non-target species of ecological and agronomic interest, such as earthworms. Earthworms are considered useful indicators of soil contamination. Exposure of these organisms to contaminants occurs mainly through the large amount of soil ingested, which passes through the digestive tract, which is closely associated with the coelom and its fluids. In this work, we used the coelomic fluids of earthworms exposed to copper sulfate and chlorpyrifos to standardize a set of non-destructive biomarkers useful for assessing the contamination in agroecosystems. Metallothionein concentrations, acetylcholinesterase inhibition, lysosomal membrane stability, micronucleus frequency, morphometric alterations, and granulocyte cytoskeleton polymerization were analyzed. The results showed that all the biomarkers used were detectable in the coelomic fluid. Furthermore, the data obtained showed highly significant variations for all biomarkers studied, thus demonstrating that the use of coelomic fluid for biomarker assessment in non-target species offers numerous advantages for field applications.

Effect of Age, Hot Beverages and Tobacco Related Products on Buccal Epithelial Cells of Cigarette Smokers and non-Smokers in Ajman, UAE.

This study aimed to find out the effect of age, hot beverages and tobacco related products on buccal mucosa cells between cigarette smokers and non-smokers in Ajman, UAE. A total of 122 samples were collected, with demographic data including age, hot beverage consumption, cigarette smoking and other tobacco practice using pre-designed questionnaires. Buccal cells were collected, stained, and screened for micronuclei (MN) under a microscope and two evaluators independently assessed all the slides. Among the 122 participants, 61.5% were aged ≤35 years, and 38.5% were aged >35 years. All non-smokers had MN values <10, while 87% of smokers had MN values >10 (p<0.001), with a trend of dose-dependent relationship between cigarette consumption and MN frequency. Similar patterns were observed in individuals using other forms of tobacco, with 97.4% exhibiting MN values >10 (p<0.001). Hot beverage consumption ≥7 cups/day was associated with 87% of subjects having MN values >10, highlighting the pattern of alternative forms of tobacco and high consumption of hot beverages association with elevated MN occurrence. Significant associations were found between MN and variables, except for age. The findings underscore the significance of tobacco and hot beverage consumption in MN occurrence, emphasizing the need to address these behaviors to mitigate genotoxicity and associated health risks. Despite age showing no significant correlation with MN frequency within the studied age range, aging combined with cigarette smoking amplifies genetic damage.<br />.

Dose-Related Mutagenic and Clastogenic Effects of Benzo[b]fluoranthene in Mouse Somatic Tissues Detected by Duplex Sequencing and the Micronucleus Assay.

Polycyclic aromatic hydrocarbons (PAHs) are common environmental pollutants that originate from the incomplete combustion of organic materials. We investigated the clastogenicity and mutagenicity of benzo[b]fluoranthene (BbF), one of 16 priority PAHs, in MutaMouse males after a 28 day oral exposure. BbF causes robust dose-dependent increases in micronucleus frequency in peripheral blood, indicative of chromosome damage. Duplex sequencing (DS), an error-corrected sequencing technology, reveals that BbF induces dose-dependent increases in mutation frequencies in bone marrow (BM) and liver. Mutagenicity is increased in intergenic relative to genic regions, suggesting a role for transcription-coupled repair of BbF-induced DNA damage. At higher doses, the maximum mutagenic response to BbF is higher in liver, which has a lower mitotic index but higher metabolic capacity than BM; however, mutagenic potency is comparable between the two tissues. BbF induces primarily C:G > A:T mutations, followed by C:G > T:A and C:G > G:C, indicating that BbF metabolites mainly target guanines and cytosines. The mutation spectrum of BbF correlates with cancer mutational signatures associated with tobacco exposure, supporting its contribution to the carcinogenicity of combustion-derived PAHs in humans. Overall, BbF's mutagenic effects are similar to benzo[a]pyrene, a well-studied mutagenic PAH. Our work showcases the utility of DS for effective mutagenicity assessment of environmental pollutants.

Genotoxic and Cytotoxic Effects of Nanoparticle and Bulk Forms of Molybdenum Trioxide and Molybdenum Disulfide Used in Bioimaging and Cancer Therapy

Nanoparticles (NPs) and bulk forms of MoO3 and MoS2 (0.1, 1, 10 and 100 µg/mL) used in cancer treatment and bioimaging were investigated by chromosome aberration, CBMN-Cyt and comet assay in human lymphocytes for the first time. This study compared both MoO3 and MoS2 and their NPs (two-dimensional (2D)) and bulk forms. Both NP and bulk forms of MoO3 and MoS2 did not cause an increase in abnormal cell frequency and CA/Cell compared to the control. While both NPs and bulk forms of MoS2 significantly increased the micronucleus frequency, MoO3 did not cause an increase. This increase was slightly higher in MoS2 NPs than in their bulk form. According to our comet assay results, both NPs and bulk forms of the MoO3 and MoS2 significantly increased the DNA damage at all concentrations. Both MoO3 and MoS2 significantly decreased MI. Neither MoO3 nor MoS2 caused a significant variation in NDI, CBPI, % cytostasis, NPB, and NBUD frequency compared to the negative control. Both particles were also characterized physicochemically. Our results revealed that MoO3 and MoS2 may have weak genotoxic and cytotoxic effects. Therefore, the toxicity potential of these particles and their underlying mechanisms for safer usage need to be investigated in more detail by other in vivo and in vitro genotoxicity and cytotoxicity tests.

Exfoliated Oral Mucosal Cell Micronucleus Frequency Evaluation in Healthy Individuals, Tobacco Users, and Patients with Potentially Malignant Disorders: An Analytical Comparative Study

ABSTRACT Objective: Assessing the prevalence of exfoliated oral mucosal cell micronuclei in tobacco smokers, healthy persons, and patients with potentially malignant diseases (PMDs). A critical comparative investigation. Materials and Procedures: The study included 80 participants divided into four groups: healthy persons, tobacco users without any clinically noticeable oral lesions, patients with leukoplakia, and patients with oral submucous fibrosis (OSMF). Each group included 20 participants. Only cases with a verified histological diagnosis of PMDs were considered for evaluating micronuclei (MNi). This included non-smokers with mucosa that seems normal, smokers with mucosa that appears normal, and individuals with leukoplakia and OSMF who were histologically identified. Results: Compared to controls, smokers and patients with PMDs had much higher MN frequencies, with PMD patients having the highest MN frequency. There was a noticeable rise in MN frequency in the H and E and PAP stains from controls to tobacco users and then to PMD patients, showing a stepwise development. While the difference between tobacco users and PMD patients was only significant with the H and E stain, the inter-group comparisons show significant differences in MN frequency between controls and both tobacco users and PMD patients across both staining techniques. Additionally, the comparison of the H and E and PAP stains reveals that whereas MN frequencies in smokers and controls are typically similar, PMD patients have considerably higher MN frequencies in the H and E stains. Conclusion: In summary, the research indicates that tobacco users have a much higher MN frequency than controls, with an even more pronounced in individuals who have PMDs.

A study on the effect of Hypericum perforatum L. extract on vanadium toxicity in Allium cepa L.

The growth of industrialization growth the risk of vanadium (V) contamination. The objective of this study was to examine the impact of 200 µg L− 1 VCI3 -induced toxicity as well as the potential protective effect of 187.5 mg L− 1 and 375 mg L− 1Hypericum perforatum (H. perforatum) extracts against this toxicity on the Allium cepa (A. cepa) model organism. For this purpose, a series of investigations were conducted on the growth physiology alterations (germination percentage, root elongation, weight gain), cytogenetic alterations (mitotic index, micronucleus, chromosomal aberrations), biochemical alterations (malondialdehyde, superoxide dismutase, catalase) and defects in meristematic tissue in A. cepa. In addition, the phenolic compound content of H. perforatum extract was determined by the LC/MS-MS method. V application negatively affected all the investigated parameters and caused a serious phytotoxic and genotoxic effect as well as oxidative stress in A. cepa. Conversely, no statistical difference was observed between the parameters of the groups treated with H. perforatum extract and those of the control group. The administration of H. perforatum extract combined with V resulted in a notable enhancement in germination percentage, root elongation, weight gain, mitotic index value, chlorophyll a level and chlorophyll b level. Additionally, it led to a reduction in micronucleus and chromosomal aberrations frequencies, as well as meristematic tissue defects. Furthermore, LC/MS-MS analysis demonstrated that H. perforatum extract contains phenolic compounds, including catechin, epicatechin, quercetin, oleuropein and rutin, which confer antioxidant properties to the extract. The study provided clear evidence that H. perforatum extract attenuates the toxic effects of V in A. cepa, which can be attributed to its high content of bioactive phenols. The findings of the study indicate that H. perforatum extract may serve as a protective natural agent for daily use against heavy metal toxicity.

Effects of Short-Term Treatment with α-Lipoic Acid on Neuropathic Pain and Biomarkers of DNA Damage in Patients with Diabetes Mellitus.

Diabetes mellitus (DM) is a complex and heterogenous disease classified as a group of metabolic disorders characterized by chronic hyperglycemia resulting from defects in insulin secretion, insulin action, or both. It leads to various complications, some of which are macrovascular or microvascular complications, like diabetic polyneuropathy (DPN), having a profound impact on patients' quality of life. Oxidative stress (OS) is one of the significant mechanisms in the development and progression of DPN. Thus, targeting OS pathways by antioxidants, such as α-lipoic acid (ALA), could represent a promising therapeutic strategy for alleviating neuropathic symptoms. The aim of our study was to evaluate whether short-term (from 4 to 9 days) intravenous administration of ALA could cause any measurable improvement in subjects with DM. Sixteen subjects with DM (six type 1 and ten type 2) and sixteen nondiabetic subjects matched by sex and age were recruited to this study. Only subjects with DM received treatment with ALA (600 mg daily). Pain intensity and biomarkers of DNA damage including plasma concentration of 8-hydroxy-2'-deoxyguanosine (8-OHdG), frequency of micronucleated lymphocytes (MN), and frequency of sister-chromatid exchanges (SCEs), were measured before and after the treatment with ALA. Pain intensity and 8-OHdG levels were significantly lower in DM subjects after the ALA treatment than before the treatment. However, no changes in the frequency of SCEs and MN were observed. Our results show some evidence that even a short-term intravenous treatment with ALA could be beneficial for diabetic subjects, reducing pain intensity and concentration of 8-OHdG in blood plasma.

Effect of exogenous application of L-mimosine on physiological, cytogenetic, biochemical and anatomical characteristics of Allium cepa L

Mimosine [β-(3‑hydroxy-4-pyridone-1-yl)-l-alanine] is a toxic non- protein amino acid, and one of its major effects is to suppress plant growth and development. Since mimosine is a compound responsible for many interesting biological activities, intensive studies have begun to be carried out on it in recent years. The current work investigated toxic effects of exogenously given 1, 2 and 3 mM l-mimosine doses on some physiological, cytogenetic, biochemical, and anatomical parameters of Allium cepa L. bulbs. For this purpose, the germination percentage (GP), root length (RL), root number (RN), and fresh weight (FW) were determinated as physiological parameters to be examined experimentally; the micronucleus (MN) frequency, chromosomal aberrations (CAs), and mitotic index (MI) were chosen as cytogenetic parameters; and the catalase (CAT) and superoxide dismutase (SOD) activities, malondialdehyde (MDA) level, and free proline (PR) content were determinated as biochemical parameters to be investigated. In addition, the changes in the root anatomical structure of the bulbs were examined under the microscope by taking cross-sections. Onion bulbs were divided into four groups as three treatments and one control (C). For seven days, the bulbs in the treatment groups were germinated with three different doses of l-mimosine (1, 2 and 3 mM), while the bulbs in the C group were germinated with tap water. Consequently, all three doses of l-mimosine caused a decrease (p < 0.05) in all investigated physiological parameter (GP, RL, RN and FW) values compared to C. Furthermore, every l-mimosine dosage resulted in a reduce (p < 0.05) in MI and an increase (p < 0.05) in MN and CA frequency compared to C group. l-mimosine application promoted CAs such as irregular mitosis, stick chromosome, lagging chromosome, protruded out chromosome, pole deviation and alignment in root meristem cells. Exposure to l-mimosine resulted in dose-related increases (p < 0.05) in SOD and CAT enzyme activity as well as MDA and PR levels compared to C group, indicating that l-mimosine also induced toxicity by causing oxidative stress in cells. Also, especially exposure to a dose of 3 mM l-mimosine caused the anatomical damages such as deformations of the epidermis and cortex cells, necrosis, buildup of certain chemical compounds in the cortex cells, thickening of the cortex cell wall, formation of vacuoles in the cell nuclei and flattened cell nuclei in the root tip meristem cells. In summary, since l-mimosine showed an inhibitory effect in the Allium cepa L. test material, it was concluded that the compound induced multi-dimensional toxicity, and the Allium test proved to be a highly valuable tool in identifying this toxicity.

Abstract B073: Leveraging micronuclei DNA from erythrocytes for early detection of hepatocellular carcinoma

Abstract Background: Hepatocellular carcinoma (HCC) is the most common form of liver cancer and accounts for ∼90% of cases globally. Detection of early-stage HCC is vital as it allows for potentially curative treatment, yet remains a significant challenge. Micronuclei (MN) are a hallmark of genomic instability. An increased frequency of MN is commonly observed in tumor cells as well as other somatic cells, including erythrocytes, in cancer patients. Here, we have established a technique (WO2021/228246 A1), for the isolation and analysis of micronuclei DNA (MN-DNA) in erythrocytes from peripheral blood. Significant changes in read densities at specific genomic locations within MN-DNA were identified in patients, termed as tumor- associated MN-DNA (taMN-DNA) features. This study evaluates the potential of these taMN- DNA features for early-stage HCC detection across human and murine model. Methods: To explore the potential of MN-DNA in cancer detection, we first collected 1-2 mL of whole blood from HDs (N = 53) and HCC patients (N = 53). MN-DNA was isolated and purified from erythrocytes, followed by whole-genome sequencing. Participants were randomly divided into training, test cohorts in an 8:2 ratio. We applied machine learning algorithms to leverage these features for cancer detection in the human model. Additionally, to investigate the formation of taMN-DNA features, we employed a well-established mouse model that mimics HCC development. Mice were injected with the genotoxic agent diethylnitrosamine (DEN) 2 weeks after birth, and developed malignant macroscopic liver nodules at approximately 40 weeks of age. Results: In the human cohort, we enrolled 106 participants, with more than half of the HCC cases were diagnosed at early-stage (stage 0-II). The cancer detection model utilizing taMN- DNA features achieved an overall accuracy of 86.3%, with a sensitivity of 81.8% and specificity of 90.9%. For early-stage, the model demonstrated sensitivities of 54.5% at a specificity of 90.9%. In the DEN-induced HCC mouse model, unsupervised hierarchical clustering based on these selected taMN-DNA features clearly separated WT and tumor mice into two distinct groups. Notably, mice that were not tumorigenic at 36 weeks post-treatment in the DEN-treated group fell into the WT group when classified by the same taMN-DNA features, indicating that the formation of taMN-DNA signatures is associated with the presence of tumors specifically. Conclusions: This pilot study highlights the potential of MN-DNA as a promising tool for early HCC detection. Leveraging these taMN-DNA features can accurately distinguish early-stage HCC patients from HDs with high sensitivity. In both human and murine models, there is a significant relationship between these signatures and tumor presence, suggesting that taMN- DNA features might reflect the diseased state across mammalian species. Research sponsor: Timing Biotech. Citation Format: Hui Lin, Haobo Sun, Xingyun Yao, Xiaoxiao Fan, Fei Meng, Honghao Liang, Xiaofei Gao. Leveraging micronuclei DNA from erythrocytes for early detection of hepatocellular carcinoma [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr B073.

Abstract A041: Detection of early-stage gastrointestinal cancers using micronuclei DNA from erythrocytes

Abstract Background: Gastrointestinal (GI) cancers account for over a third of cancer deaths. However, their early detection remains a challenge. Micronuclei (MN) are cytoplasm-resident subcellular structures, containing damaged chromosome segments that indicative of genomic instability. Increased MN frequency in hematopoietic cells is observed in patients with solid tumors. Our developed technique (WO2021/228246 A1) enables the purification and characterization of micronuclei DNA in erythrocytes from peripheral blood. By comparing MN-DNA from healthy donors (HDs) and GI cancer patients, we identified significant changes in read densities at specific genomic locations in patients, which were termed as tumor-associated MN-DNA (taMN- DNA) features. Here, we evaluated the potential of these features for early-stage GI cancer detection, including colorectal (CC), gastric (GC) and esophageal (EC) cancers. Methods: Peripheral blood (1-2 mL) was collected from healthy donors (HD) and cancer patients MN-DNA isolation and purification from erythrocytes. Participants were randomly divided into training, validation and independent test cohorts in a 7:2:1 ratio, maintaining similar cancer types, gender and age distribution. Distinct MN-DNA features between cancer patients and HDs were identified using sequencing data. Machine learning algorithms were applied using these features for cancer detection. Results: The study enrolled 1753 participants, including 987 HDs, 420 CC, 282 GC and 64 EC cases. Of these, over half were diagnosed with early-stage diseases; TNM stage 0-I accounted for 40.57%, stage II for 23.32%, stage III for 29.25% and stage IV for 6.87%. The pan-cancer model achieved 88.2% sensitivity with an overall specificity of 95.4%. Patients in individual caner types were detected from the tissue of origin classification with an overall accuracy of 91.9% based on pan-cancer at 95.4% specificity. Sensitivity was 90.8% for CC, 91.7% for GC and 100% for EC. For early- stage (stage 0-II) CC, GC and EC, the model demonstrated sensitivities of 97.4%, 94.7% and 100.00%, respectively, at 95.4% specificity. Conclusions: Unlike cell-free DNA, MN-DNA are chromosomal fragments in the cytoplasm. The abundance of erythrocytes in peripheral blood provides an easily accessible source for MN-DNA enrichment. This pilot study illustrates the potential of MN-DNA as a tool for early GI cancer detection, contributing to large-scale efforts towards developing an effective GI cancer screening test. Research sponsor: Timing Biotech. Citation Format: Haobo Sun, Xingyun Yao, Yuehua Han, Yurong Jiao, Xiangxing Kong, Chengcheng Liu, Qingqu Guo, Fei Meng, Honghao Liang, Hongbo Li, Xiuqin Fan, Jun Li, Xiaofei Gao. Detection of early-stage gastrointestinal cancers using micronuclei DNA from erythrocytes [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A041.

Novelties on tradescantia: Perspectives on water quality monitoring.

In the face of intense urban expansion, the assessment of water quality plays a crucial role in environmental preservation. Here, we evaluated aquatic genotoxicity in three locations with different degrees of urbanization using Tradescantia pallida var. purpurea and Daphnia magna as bioindicators. The objective was to investigate the influence of urbanization on water quality and the efficiency of the TRAD-MCN biological test in monitoring aquatic genotoxicity. Therefore, we established the genotoxic potential by evaluating micronucleus frequency in T. pallida and immobilization and DNA damage in the standard test with D. magna during two seasons of the year (dry and rainy). Our results showed that the frequency of micronuclei in T. pallida (TRAD-MCN) was significantly higher in the locations with a higher degree of urbanization, independently of the seasons. The tests with D. magna revealed a higher rate of immobilization and DNA damage in the location most impacted by residential and industrial effluents (especially mining activities). Additionally, the TRAD-MCN proved to be equivalent to the standard test for genotoxicity assessment, supporting its potential applicability in environmental monitoring. Finally, we observed that urbanization significantly influences water quality, and among the evaluated physicochemical parameters, dissolved oxygen was shown to be the most important driver of the water quality index (WQI). Our findings have significant implications for water resource management, underlining the need for policies that consider the specificities of different regions. This highlights the robustness, flexibility, and reliability of T. pallida as an environmental monitoring tool.

Genotoxic Damage in Rainbow Trout Oncorhynchus Mykiss Exposed to Transport Stress

Transporting live fish is a common technique in the aquaculture industry. This research examined how 3-hour transportation stress affects the micronucleus frequency of rainbow trout (Oncorhynchus mykiss). The micronucleus test was used to assess micronuclei and nuclear abnormalities in peripheral erythrocytes. Fish were sampled before (control) and immediately after the 3-hour transport process (t0 group), 6 hours after the transport process (t6 group), 12 hours after the transport process (t12 group), and 24 hours after the transport process (t24). The research found that the greatest MN frequency was substantially detected in the t0 group (p