Frequency Of Micronuclei Research Articles

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

Background/Objectives: 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. Methods: 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. Results: 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. Conclusions: 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.

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.

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

Genotoxic effects in island populations of Cartagena de Indias, Colombia due to environmental exposure to mercury and cadmium

Background Contamination of Cartagena Bay, Colombia with heavy metals such as mercury (Hg) and cadmium (Cd) presents a major environmental and public health concern, particularly for human communities residing on nearby islands and coastal areas. These populations face enhanced exposure risks owing to their traditional fishing practices and continuous interactions with polluted marine environments. This study aimed to evaluate the genotoxic effects of environmental exposure to Hg and Cd in populations from the island zone of the Cartagena district, Bolívar. Methods Ninety-four individuals from the four island communities (study group) and 30 individuals from the urban area of Cartagena (control group) participated in this study. The blood samples were collected to measure total mercury (T-Hg) and Cd concentrations, and a Buccal Micronucleus Cytome (BMCyt) assay was used to evaluate exposure effects. Results Cadmiun levels in the blood of the study group were within the normal range and comparable to those of the control group (p > 0.05). However, the study group exhibited significantly higher T-Hg levels (7.34 μg/L) compared to the control group (2.01 μg/L), surpassing the accepted limit. Moreover, the study group showed a higher incidence of DNA damage and cell death biomarkers (p < 0.05). Additionally, significant correlations were observed between total blood Hg levels and the frequencies of micronuclei, karyorrhexis, and karyolysis. Conclusion These results suggest that island populations of Cartagena are exposed to high levels of Hg and exhibit genotoxic damage, indicating a problem that must be addressed.

FDXR gene expression and micronucleus frequency in Type 2 diabetes mellitus patients and their importance in case of radiation exposures

Analysis of gene expression (e.g. Ferredoxin Reductase: FDXR) changes in the blood samples have shown potential as predictive markers of disease, prognosis for therapy as well as triage and dose estimation in radiation-exposed populations. Similarly, quantification of micronuclei (MN) formation has been presented as a rapid cytogenetic marker for those applications. It was cautioned that the reliable utilization of these markers for prediction of disease, therapy prognosis, and dose estimation depends upon the information on known variables that affect these markers. Literature suggests that advanced glycation end products/oxidative stress in diabetic conditions can alter the levels of DNA damage and gene expression, and impact the segregation of the exposed from unexposed during nuclear disasters. Therefore, we investigated the influence of Type 2 diabetes mellitus (T2DM) on baseline expression of the FDXR and frequency of MN. Peripheral blood samples were collected from healthy volunteers (HV) (n = 60; 43 males and 17 females) and T2DM patients (n = 60; 32 males and 28 females), and performed real-time quantification of FDXR gene expression and analysis of MN frequency using microscopy. The basal level of FDXR gene expression (2.55 folds) (p < 0.01) and the frequency of MN is significantly (p < 0.01) higher (4 folds) in T2DM patients when compared to HV. Further, subgroup analysis found that gender, alcohol, smoking, duration of T2DM, complications, and medications increased both the expression of the FDXR gene and frequency of MN in T2DM; nevertheless, the increase was not significant, except for gender (p < 0.05) and medication (p < 0.05) on the frequency of MN. Overall results indicate that the T2DM patients showed a higher basal level expression of the FDXR gene and MN frequency when compared to HV and suggest an altered metabolic condition in T2DM is a confounding factor that impacts the levels of those markers. The increased levels of these markers might need to be considered to monitor medical radiation exposures and reliable biodosimetry during large-scale radiological accidents.

DNA Damage, Cell Death, and Alteration of Cell Proliferation Insights Caused by Copper Oxide Nanoparticles Using a Plant-Based Model.

The speedy growth of copper oxide nanoparticle (CuO NP) manufacturing due to their wide application in industries has caused concerns due to their increased discharge into the environment from both purposeful and accidental sources. Their presence at an elevated concentration in the environment can cause potential hazards to the plant kingdom, specifically to staple food crops. However, limited research is available to determine the consequences of CuO NPs. The present study aimed to assess the morphological and cytological changes induced by CuO NPs on Pisum sativum L., a key staple food crop. Seeds of Pisum sativum were exposed to various concentrations of CuO NPs (0, 25, 50, 75, 100, and 125 ppm) for 2 h, and their effects on seed germination (SG), radicle length (RL), cell proliferation kinetics (CPK), mitotic index (MI), cell death (CD), micronucleus frequency (MNF), and chromosomal aberration frequency (CAF) were studied. The results indicate a significant reduction in SG, RL, CPK, and MI and a significant dose-dependent increase in CD, MNF, and CAF. CuO NP treatment has led to abnormal meiotic cell division, increased incidence of micronucleus frequency, and chromosomal aberration frequency. Additionally, the CuO NP-treated groups showed an increase in the percentage of aberrant meiotic cells such as laggard (LG), double bridge (DB), stickiness (STC), clumped nuclei (CNi), precocious separation (PS), single bridge (SB), and secondary association (SA). CuO NP treatment led to reductions in SG as follows: 55% at 24 h, 60.10% at 48 h, and 65% at 72 h; reductions in RL as follows: 0.55 ± 0.021 cm at 24 h, 0.67 ± 0.01 cm at 48 h, and 0.99 ± 0.02 cm at 72 h; reductions in CPK as follows: 34.98% at prophase, 7.90% at metaphase, 3.5% at anaphase, and 0.97% at telophase. It also led to a 57.45% increase in CD, a 39.87% reduction in MI, and a 60.77% increase in MNF at a higher concentration of 125 ppm. The findings of this study clearly show that CuO NPs have a genotoxic effect on the food crop plant Pisum sativum.

Genotoxicity Of The Herbicide Picloram (TORDON ® 24K) on tadpoles of Boana pulchella (Amphibia, Anura, Hylidae)

Picloram (PIC) is an herbicide widely used in Uruguay to control weeds, especially in forest crops. In the present study, we evaluate the acute toxicity and genotoxicity of formulated 24.9% PIC-based Tordon®24K on Boana pulchella tadpoles under standard laboratory conditions. In the acute toxicity test we use mortality as the endpoint and we obtained a 48-h LC50 of 16.5 mg/L, LOEC 14.5 mg/L, and NOEC 12.5 mg/L. We determined genotoxicity in GS36 tadpoles exposed to sublethal concentrations of 8.5 (NOEC/1.5) and 12.5 mg/L (NOEC) PIC using micronucleus frequency (MN) and occurrence of nuclear abnormalities (AN) as endpoints. We noticed a significant increase in the frequency of MN and AN in all treatments tested at sublethal concentrations. Our findings demonstrate the genotoxic effects of this herbicide, which represent a danger to amphibians that inhabit agroecosystems. Based on the results obtained and considering that Boana pulchella is not a threatened species and has a wide distribution in the Uruguayan territory and the Pampa Biome, it could be considered a reference organism in environmental monitoring programs in the region.

In Vitro and In Vivo Genotoxicity of Polystyrene Microplastics: Evaluation of a Possible Synergistic Action with Bisphenol A.

The ubiquitous presence of plastics represents a global threat for all ecosystems and human health. In this study, we evaluated, in vitro and in vivo, the genotoxic potential of different concentrations of polystyrene microplastics (PS-MPs) and their possible synergistic interactions with bisphenol-A (BPA). For the in vitro and the in vivo assays, we used human lymphocytes and hemocytes from Lymnaea stagnalis, respectively. The genomic damage was evaluated by the micronucleus assay, and differences in eggs laid and growth of L. stagnalis were also evaluated. In human lymphocytes, PS-MPs alone at the concentration of 200 μg/mL and in association with BPA 0.100 µg/mL significantly increased the frequencies of micronuclei and nuclear buds, indicating a possible in vitro genotoxic additive action of these two compounds. Vice versa, PS-MPs did not result in genotoxicity in hemocytes. Our results indicated that PS-MPs have genotoxic properties only in vitro and at a concentration of 200 µg/mL; moreover, this compound could intensify the genomic damage when tested with BPA, indicating possible cumulative effects. Finally, PS significantly reduced the growth and the number of laid eggs in L. stagnalis.

Toxicity of paclobutrazol-based pesticide on Lactuca sativa L.: germination, seedling development, and DNA damage.

Paclobutrazol, a fungicide of the triazole class, is widely used as an inducer of early flowering and fruiting by inhibiting gibberellin formation. However, biological assays using model organisms to evaluate their cytogenotoxic and mutagenic potential are still scarce. Therefore, this study aimed to investigate the effects of the commercial product Cultar® 250 SC (CP) and the pure substance (PBZ) on the germination and initial seedling development of Lactuca sativa L. (lettuce), in addition to evaluating the effects of CP on the mitotic activity and DNA, as we believe that PBZ has a greater toxic potential than CP on seed germination, and that the latter has cytogenotoxic and mutagenic effects on L. sativa. Lettuce seeds treated with CP and with PBZ in the doses of 0.25, 0.50, 1, 1.5, and 2g L-1 showed significant reductions in germination rate, as well the CP reduced the root and initial development seedling development. PBZ showed greater inhibition of germination compared to CP. In direct exposure to PBZ, there was not sufficient seedling development for analysis, while in discontinuous treatment, there was inhibition of root growth (except for doses of 0.25 and 0.50g L-1) and in the development of the aerial part. While no mitodepressive effect was observed in meristematic cells treated with CP, increased frequencies of chromosomal alterations, including condensed nuclei and micronuclei, were evident in both meristematic cells and the F1 region. The Comet assay further demonstrated higher levels of DNA damage at higher paclobutrazol doses, supporting the findings of increased micronucleus frequencies. Consequently, it can be concluded that the CP exhibits greater toxicity towards seed germination compared to lettuce seedlings, and PBZ has a greater toxic potential than CP in relation to these parameters. However, the impact of CP on seedlings was relatively minimal, as evidenced by their limited effects on development, cell proliferation, and DNA, suggesting a slight toxicity of this agent. Therefore, we infer that Cultar® 250 SC should be used with caution. Thus, this study emphasizes the importance of employing joint analyses to better elucidate and correlate the mechanisms of action of potentially toxic substances. Furthermore, it provides a basis for discussing the application of Cultar® 250 SC and seeking more sustainable alternatives in food production.

Investigation of genetic instability in patients with Diabetes Mellitus type I, II and LADA using buccal micronucleus cytome assay

The aim of our pilot study was to investigate the frequency of micronuclei (MN) and other nuclear anomalies in exfoliated cells of the oral mucosa in patients with type I, II, and LADA (Latent Autoimmune Diabetes in Adults, classified as type 1.5 intermediate, slowly progressing diabetes) types of diabetes mellitus (DM) and compare them with healthy individuals of the Armenian population using the MN test. For each participant essential clinical and biochemical parameters were studied, including blood pressure, duration of illness, glycosylated hemoglobin (HbA1c), blood glucose, plasma glucose, urea, total protein, creatinine, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, HOMA-IR (insulin resistance), insulin, and triglycerides, as well as necessary anthropometric, genealogical, and genetic data. All participants were surveyed regarding habits that might affect MN levels, such as smoking, alcohol consumption, drug use, hereditary diseases, and viral infections. Cytogenetic analyses of exfoliated cells showed that the level of MN in exfoliated cells of DM patients was elevated approximately two to three times compared to healthy individuals. However, statistical significance was only reached in type I DM and LADA patients. The levels of other nuclear anomalies in the squamous epithelial cells of DM patients were also analyzed, and a significant increase in their levels was observed in all three DM types, indicating cytotoxic and genotoxic effects. The results of this study also revealed a high correlation between the total number of MN, cells with MN, blood glucose concentration, and glycosylated hemoglobin.

Herbicide paraquat dichloride augments the expression of the c-abl-noxa-apoptotic pathway, instigating ovarian atresia in edible fish, Channa punctatus.

Ovarian cells of animals undergo atresia to regulate egg numbers and their release. However, unrestrained atretic activity decreases the chances of oocyte maturity and further process, ultimately resulting in a reduced population. Edible fishes are a critical source of nutrition and it is required to regulate their population. The application of herbicides at fish production sites is a threat to their population by altering female reproductive activity. In concern to this, here, this 60-day study investigates herbicide paraquat dichloride (PD)-induced atresia in ovarian cells of Channa punctatus (C. punctatus). Further, 96-h LC50 of PD for C. punctatus was calculated as 58.40mg/L. At the end of each exposure period, an increase in intracellular ROS in blood cells and activities of SOD and CAT in ovary tissue were observed in PD-treated fish with increasing concentration. The frequency of micronuclei in blood cells subsequently increased with the highest PD concentration. A significant (p < 0.05) increase in the relative expression of target genes was observed at the 30th and 60th day of the sampling periods in the fish treated with the highest concentration of PD. The histological study confirmed the occurrence of atretic ovary cells in PD-exposed fish. From the results it can be concluded that PD enhances the uncontrolled death of ovarian cells and has consequences of impairing the ovary health and reproductive potential of fish, hence reducing the fish population.

QBD approach for green synthesis of Rutin silver nanoparticles- screening for antioxidant, anticancer and anticlastogenic potential

Rutin is a flavonoid glycoside abundant in many plants exhibiting pharmacological activities like antioxidant, anticancer, anti-inflammatory and antimicrobial activities. Plant biomarkers suffer low bioavailability and solubility that lack clinical effectiveness. The smart nanoparticles conversion addresses this limitation with optimal particle size and targeted drug delivery. The present study involves QbD approach for formulation of Rutin silver nanoparticles and evaluation of antioxidant, anticancer and anticlastogenic potential. QbD experimentation involved particle size and drug release as dependent variables over the silver nitrate concentration, methanol and sonication time as independent variables devising 15 formulations (F1 –F15). F12 formulation was found to be optimized with 126.3 nm average size, stable and dispersible characterized by UV, FTIR, SEM and DLS studies. The calibration curve of Rutin was plotted at 352 nm with linearity (LOD = 0.061 μg/ml and LOQ = 0.187 μg/ml). The invitro drug release studies by USP dissolution apparatus I (Basket type) proved the sustained release characteristics with 97.3 % drug release when compared to the Rutin. The pharmacological screening for potential antioxidant and anticancer activity on G361 and MCF 7 cell line of F12 formulation have shown promising results and also enhanced solubility in water compared to Rutin. Anticalstogenic potential as a function of induced micronuclei frequency was evaluated as a characteristic feature in bone marrow cells obtained from mice. Results indicate pre-treatment with the F12 reduced frequency of micronuclei in mouse bone marrow cells caused by Cyclophosphamide (CP) significantly. The protective effect of F12 in suppression was demonstrated at both dosages of 100 and 200 mg/kg. Thus the findings suggest the novel Rutin silver nanoparticles as lead drug serving as antioxidant, anticancer and anticlastogenic agent.

Impact of graded doses of enrofloxacin on the safety and biological responses of Nile tilapia Oreochromis niloticus

The cultivation of tilapias, the third most farmed fish group globally, has been rapidly growing, especially in Southeast Asia. This surge in tilapia farming intensification has led to increased use of antibiotics to control bacterial diseases. This study investigated the safety implications of administering graded doses of enrofloxacin (ENF) at 0 (control), 10, 30, 50 and 100 mg/kg biomass/day orally to Oreochromis niloticus. The 43-day study comprised 7 days of pre-dosing, 15 days of ENF-dosing, and a 21-day recovery period with a periodical assessment of the biological responses of fish. The results revealed that the overdosed groups experienced up to 21% reduction in feed consumption, 11% mortalities, and adverse impacts on hematology, including a decrease in erythrocytes, and monocytes and an increase in leukocytes, thrombocytes, lymphocytes, and neutrophils. Haematological indices like mean corpuscular volume and mean corpuscular hemoglobin decreased, while mean corpuscular hemoglobin concentration increased. The plasma biochemical parameters including glucose and liver and kidney enzymes unveiled a significant dose- and time-dependent increase, while calcium and chloride levels decreased. Erythrocytes displayed several erythrocyte cellular and nuclear abnormalities. The frequency of micronucleus increased with dose and time, suggesting potential genotoxicity of ENF. Additionally, a dose-dependent increase in residues in the tissues with the highest accumulation in muscle was documented. Nevertheless, the recovery of the measured parameters upon dose termination indicated that the ENF-induced alterations are reversible. The study affirmed the safety of ENF at the recommended dose (10 mg) in O. niloticus and their adoptive responses to higher doses.

Isolation and comparative genotoxicity screening of trichokonins VI and VIII on CHO-K1 cells

Peptaibols are fungal peptides that exhibit efficacy against pathogen microorganisms. Trichokonin VI (TK-VI) and trichokonin VIII (TK-VIII) are known peptaibols isolated from the endolichenic fungi Hypocrea sp. Previous investigations reported that trichokonin VI presents antiproliferative effects on tumor cells. This study is pioneering in elucidating the genotoxic effects of TK-VI and TK-VIII, contributing to the thorough assessment of their safety as potential therapeutic agents. The present investigation aimed to evaluate the genotoxicity of TK-VI and TK-VIII on CHO-K1 cells. Cytotoxicity was evaluated using the XTT assay and clonogenic survival assays, followed by evaluation of DNA damage using the comet assay and micronucleus test conducted in vitro. The XTT assay results indicated IC50 values of 10.30 µM and 9.89 µM for TK-VI and TK-VIII, respectively. The clonogenic survival assay indicated that concentrations of 10 µM or higher completely inhibited the cell colony formation. In the comet assay, both TK-VI and TK-VIII increased the DNA damage score and the frequency of comet nuclei in all tested concentrations. In the micronucleus assay, TK-VI and TK-VIII at 10 µM increased the frequency of MN in CHO-K1 cells. Both TK-VI and TK-VIII exhibited genotoxic effects. Our findings underscore the importance of considering the genotoxicological safety of peptaibols, particularly when assessing their potential for other biological activities.

Comparative analysis of cyto-genotoxicity of zinc using the comet assay and chromosomal abnormality test.

In this study, the toxicity of the trace element zinc (Zn) in Allium cepa L. test material was examined. Toxicity was investigated in terms of physiological, cytogenetic, biochemical, and anatomical aspects. Germination percentage, root length, weight gain, mitotic index (MI), micronucleus (MN) frequency, chromosomal abnormalities (CAs), malondialdehyde (MDA), proline and chlorophyll levels, superoxide dismutase (SOD) and catalase (CAT) enzyme activities, and meristematic cell damage were used as indicators of toxicity. Additionally, the comet test was used to measure the degree of DNA damage. Four groups of A. cepa bulbs-one for control and three for applications-were created. While the bulbs in the treatment groups were germinated with Zn at concentrations of 35, 70, and 140mg/L, the bulbs in the control group were germinated with tap water. Germination was carried out at room temperature for 72h and 144h. When the allotted time was over, the root tips and leaf samples were collected and prepared for spectrophotometric measurements and macroscopic-microscopic examinations. Consequently, Zn treatment led to significant reductions in physiological indicators such as weight gain, root length, and germination percentage. Zn exposure caused genotoxicity by decreasing the MI ratios and increasing the frequency of MN and CAs (p < 0.05). Zn promoted various types of CAs in root tip cells. The most observed of CAs was the sticky chromosome. Depending on the dose, Zn was found to cause an increase in tail lengths in comet analyses, which led to DNA damage. Exposure to Zn led to a significant decrease in chlorophyll levels and an increase in MDA and proline levels. It also promoted significant increases in SOD and CAT enzyme activities up to 70mg/L dose and statistically significant decreases at 140mg/L dose. Additionally, Zn exposure caused different types of anatomical damage. The most severe ones are epidermis and cortex cell damage. Besides, it was found that the Zn dose directly relates to all of the increases and decreases in physiological, cytogenetic, biochemical, and anatomical parameters that were seen as a result of Zn exposure. As a result, it has been determined that the Zn element, which is absolutely necessary in trace amounts for the continuation of the metabolic activities of the organisms, can cause toxicity if it reaches excessive levels.

Sex differences in constructing dose-response calibration curves for micronuclei using cytokinesis-blocked micronucleus assay for radiation biological dosimetry in the Iranian population

Biological dosimetry, using chromosome damage biomarkers, can be considered as a key measure for radiation overexposure assessment. Therefore, for accurate dose estimation through biodosimetry, it's imperative for each biological dosimetry laboratory to establish its own specific dose-response calibration curve. In this research, the cytokinesis-blocked micronucleus (CBMN) assay was utilized to determine the frequencies of micronuclei (MN) per binucleated cell in human peripheral blood lymphocytes exposed to x-ray radiation using a LINAC (6 MV) at doses up to 4 Gy. The aim was to establish an in vitro dose-response calibration curve in our laboratory for the Iranian demographic by analyzing blood samples of ten participants (5 males and 5 females) through CABAS and Dose Estimate software. Our findings indicate an over-dispersion of the Poisson distribution in the pooled data across both sexes, coupled with a linear-quadratic increase in MN yield with dose which was particularly pronounced in the female group. The constructed dose-response curves for micronuclei yield are represented by equations Y= (0.0102 ± 0.0016) +(0.0296 ± 0.0054) D+(0.0232 ± 0.0017) D2 and Y= (0.0084 ± 0.0010) +(0.0212 ± 0.0034) D+(0.0160 ± 0.0011) D2 for the female groups, respectively. The alpha and beta coefficients, derived from the Dose Estimate software for each male and female group, were closely comparable with those obtained from the CABAS program and previous studies. The analysis of statistical parameters such as the Weighted Chi Squared (χ2) and p-value suggests that using distinct curves for each sex, rather than a unified biodosimetry formula for pooled data, ensures more accurate radiation dose estimates. Consequently, the findings of this study provide us with the assurance to utilize the derived calibration curve of MN for upcoming biological dosimetry needs in Iran. However, to enhance the reliability of results, it's essential to use biodosimetry with diverse assays and consider all clinical and physical parameters, given the limitations of cytogenetic assays.

LC-MS/MS phenolic profileand remedial role ofUrtica dioica extract against Li2CO3-induced toxicity.

In this study, the protective role of Urtica dioica extract (Udex) against Li2CO3 toxicity in Allium cepa L. was investigated using various parameters such as germination rates, root growth, weight gain, mitotic index (MI), malondialdehyde (MDA), micronucleus (MN), antioxidant enzyme activity, chromosomal abnormalities (CAs) and anatomical changes. As the biological activity of Udex is related to its active content, the profile of phenolic compounds was determined by LC-MS/MS analysis. Li2CO3 caused abnormalities in the tested parameters and serious regressions in germination parameters. Application of 100mg/L Li2CO3 reduced the chlorophyll a and b contents by 73.04% and 65.7%, respectively. Li2CO3 application exhibited a cytotoxic effect by inducing significant decreases in MI and increases in the frequency of MN, and also showed a genotoxic effect by causing CAs. After 100mg/L Li2CO3 treatment, MDA, proline, superoxide dismutase, and catalase levels increased by 54.9%, 58.5%, 47.8%, and 52.3%, respectively. Li2CO3 and Udex co-administration resulted in a regression in increased biochemical parameters and genotoxicity parameters, and an improvement in germination parameters. Furthermore, Udex demonstrated efficacy in mitigating the detrimental effects of Li2CO3 on the root tip, particularly in the 200µg/mL Udex-treated group. The thickening of the cortex cell wall and conduction tissue, which is commonly induced by Li₂CO₃, was not observed in the Udex-treated group. The protective effect of Udex can be explained by the phenolic compounds it contains. Rutin was detected as the major component in Udex and other phenolics were listed according to their presence rate as protecatechuic acid > caffeic acid > p-coumaric acid > syringic acid > rosemarinic acid > epicatechin. Li ions, which increase in the environment after industrialization, are an important environmental pollutant and exhibit toxicity that affects many pathways in organisms. Scientific research should not only detect these toxic effects but also develop solutions to such problems. In this study, it was determined that the Udex application had a toxicity-reducing role against Li2CO3 toxicity. Also, it has been demonstrated that A. cepa is an important indicator in determining this toxicity and toxicity-reducing applications.

Investigation of the protective role of Ginkgo biloba L. against phytotoxicity, genotoxicity and oxidative damage induced by Trifloxystrobin

Trifloxystrobin (TFS) is a widely used strobilurin class fungicide. Ginkgo biloba L. has gained popularity due to its recognized medicinal and antioxidant properties. The aim of this study was to determine whether Ginkgo biloba L. extract (Gbex) has a protective role against TFS-induced phytotoxicity, genotoxicity and oxidative damage in A. cepa. Different groups were formed from Allium cepa L. bulbs subjected to tap water (control), 200 mg/L Gbex (Gbex1), 400 mg/L Gbex (Gbex2), 0.8 g/L TFS solution (TFS), 200 mg/L Gbex + 0.8 g/L TFS (TFS + Gbex1) and 400 mg/L Gbex + 0.8 g/L TFS (TFS + Gbex2), respectively. The phenolic composition of Gbex and alterations in the morphological, physiological, biochemical, genotoxicity and anatomical parameters were evaluated. Rutin, protocatechuic acid, catechin, gallic acid, taxifolin, p-coumaric acid, caffeic acid, epicatechin, syringic acid and quercetin were the most prevalent phenolic substances in Gbex. Rooting percentage, root elongation, weight gain, chlorophyll a and chlorophyll b decreased by approximately 50%, 85%, 77%, 55% and 70%, respectively, as a result of TFS treatment compared to the control. In the TFS group, the mitotic index fell by 28% compared to the control group, but chromosomal abnormalities, micronuclei frequency and tail DNA percentage increased. Fragment, vagrant chromosome, sticky chromosome, uneven chromatin distribution, bridge, vacuole-containing nucleus, reverse polarization and irregular mitosis were the chromosomal abnormalities observed in the TFS group. The levels of proline (2.17-fold) and malondialdehyde (2.71-fold), as well as the activities of catalase (2.75-fold) and superoxide dismutase (2.03-fold) were increased by TFS in comparison to the control. TFS-provoked meristematic disorders were damaged epidermis and cortex cells, flattened cell nucleus and thickened cortex cell wall. Gbex combined with TFS relieved all these TFS-induced stress signs in a dose-dependent manner. This investigation showed that Gbex can play protective role in A. cepa against the phytotoxicity, genotoxicity and oxidative damage caused by TFS. The results demonstrated that Gbex had this antioxidant and antigenotoxic potential owing to its high phenolic content.

Genotoxic effects in island populations of Cartagena de Indias, Colombia due to environmental exposure to mercury and cadmium

Background Contamination of Cartagena Bay, Colombia with heavy metals such as mercury (Hg) and cadmium (Cd) presents a major environmental and public health concern, particularly for human communities residing on nearby islands and coastal areas. These populations face enhanced exposure risks owing to their traditional fishing practices and continuous interactions with polluted marine environments. This study aimed to evaluate the genotoxic effects of environmental exposure to Hg and Cd in populations from the island zone of the Cartagena district, Bolívar. Methods Ninety-four individuals from the four island communities (study group) and 30 individuals from the urban area of Cartagena (control group) participated in this study. The blood samples were collected to measure total mercury (T-Hg) and Cd concentrations, and a Buccal Micronucleus Cytome (BMCyt) assay was used to evaluate exposure effects. Results Cadmiun levels in the blood of the study group were within the normal range and comparable to those of the control group (p &gt; 0.05). However, the study group exhibited significantly higher T-Hg levels (7.34 μg/L) compared to the control group (2.01 μg/L), surpassing the accepted limit. Moreover, the study group showed a higher incidence of DNA damage and cell death biomarkers (p &lt; 0.05). Additionally, significant correlations were observed between total blood Hg levels and the frequencies of micronuclei, karyorrhexis, and karyolysis. Conclusion These results suggest that island populations of Cartagena are exposed to high levels of Hg and exhibit genotoxic damage, indicating a problem that must be addressed.