Could cannabigerol protect against neuroinflammation? Insights from an in vitro microglial study.

RAD51 promotes osteogenic differentiation and inhibits DNA damage in osteoporosis though regulating cGAS-STING signaling pathway.

FOXA2/ALDOB axis modulation of fatty acid beta-oxidation influences irinotecan resistance in colorectal cancer.

Vimentin-targeting adaptogen withaferin A: Potential to selectively suppress cervical cancer - Single-cell microspectroscopic and molecular analysis.

Bee bread as a natural shield against genetic damage: Investigation of antigenotoxic and antimutagenic potential.

Evaluation of the genotoxic potential of dietary metabolites to support the renewal of an active substance under European Commission Regulation (EC) 1107/2009 - fludioxonil case study.

Exposure to hexavalent chromium and 1800 MHz electromagnetic radiation can synergistically induce intracellular DNA damage in mouse embryonic fibroblasts.

Comprehensive toxicological profiling of Costus pictus D. Don methanolic leaf extract using in-vitro and in-vivo model.

Endometriosis is a chronic, hormone-dependent disorder characterized by ectopic implantation of endometrial tissue, often accompanied by pain and infertility. Although the progesterone receptor modulator RU486 is effective for pain relief, its impact on lesion regression is limited, possibly due to apoptosis resistance and overexpression of PARP1 in endometriotic cells. This study aimed to evaluate the synergistic therapeutic efficacy of combining RU486 with the PARP inhibitor olaparib, focusing on reactivating p53-dependent apoptosis in endometriotic lesions. Primary ectopic endometrial stromal cells (EESCs) were used to assess the effects of RU486 and olaparib on cytotoxicity, apoptosis, DNA damage, mitochondrial membrane potential, and cell migration. Mechanistic investigations included flow cytometry, western blotting, RT-qPCR, immunofluorescence, and comet assays. A murine endometriosis model was established to evaluate in vivo lesion suppression, pain behaviour, hormone levels, and treatment safety following combination therapy. RU486 and olaparib exhibited strong synergy at a 1:4 ratio, significantly enhancing apoptosis in EESCs by coactivating p53 through distinct mechanisms-transcriptional up-regulation by RU486 and posttranslational stabilization by olaparib. This synergy intensified Bax/Bcl-2-mediated mitochondrial dysfunction, caspase-3 activation, and PARP1 cleavage. In vivo, the combination therapy achieved a 77.5% lesion weight regression, significantly exceeding monotherapies, while maintaining robust analgesic effects. Histological and molecular analyses confirmed DNA damage and enhanced apoptosis in lesions. The combination medication enhanced apoptotic sensitivity in endometriosis by restoring p53 activity through targeting hormonal signalling and DNA repair pathways. This strategy offered strong potential for clinical translation.

Cancer is a major public health problem due to uncontrolled cell proliferation, invasion into surrounding tissues, and its high prevalence and mortality rates. Despite current treatment strategies, the poor prognosis in many cancer types necessitates the exploration of new pharmacological approaches. In this study, we aimed to evaluate the cytotoxic and genotoxic effects of Paroxetine, a potent selective serotonin reuptake inhibitor (SSRI), on human ovarian cancer (A2780), prostate cancer (LNCaP), and colon cancer (Caco-2) cell lines under in vitro conditions. A2780, LNCaP, and Caco-2 cells were cultured in appropriate media and treated for 24 hours with paroxetine at concentrations of 1, 10, 100, and 1000 µM. Cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-diphenyltetrazolium bromide (MTT) assay, and the inhibitory concentration was calculated using GraphPad Prism 8 software. DNA damage was assessed using the comet assay. Statistical analyses were performed with IBM SPSS Statistics 24.0 (Windows), and p

The study evaluated the genotoxicity of sodium benzoate and potassiumsorbate using the comet assay, employing erythrocytes of Oreochromis niloticus (tilapia) as an ex vivo biological model. This model is a viable alternative due to its genetic homogeneity, nucleated cells, and low cost. Treatments were conducted with concentrations of 40, 65, and 90 mg/ml of both preservatives and were compared with positive (cyclophosphamide) and negative (fetal bovine serum) controls. The results showed that potassium sorbate has a greater genotoxic effect than sodium benzoate, with a clear dose-response relationship in DNA fragmentation. From 40mg/ml, sorbate showed a significant increase in DNA damage, and at 90 mg/ml, thedamage was comparable to the positive control. Sodium benzoate presented a lesssevere effect, maintaining high cell viability at concentrations of 40 and 65 mg/ml, while sorbate reduced viability at higher concentrations, with an LD50 of 163 mg/ml. Parameters such as the percentage of tail DNA and olive tail moment reflected an increase in genetic damage induced by sorbate. These findings demonstrate thattilapia erythrocytes are an effective model for evaluating cytotoxicity and genotoxicity,highlighting the need to regulate the concentrations of food preservatives to protectgenetic integrity and consumer safety.

Abstract Background: Rlip (aka RLIP76 protein encoded by RALBP1) is a Ral-regulated, membrane-associated ATPase enzyme that facilitates Phase 2 detoxification, plays a role in clathrin-dependent endocytosis (CDE), and is vital for cancer cell formation and survival. DNA repair is essential for both normal and cancer cell survival, as it protects cancer cells from DNA damage caused by chemotherapy or radiation. Poly (ADP-ribose) polymerases (PARPs) play a crucial role in the repair of DNA damage. By targeting these repair mechanisms in cancer cells, it is possible to improve the effectiveness of cancer treatments. A notable example is dysfunctional homology-directed repair (HDR), which results from inactivating mutations in the BRCA1 and BRCA2 genes, contributing to radiation resistance in human cancer cells. Therefore, identifying new targets within double-strand break (DSB) repair pathways is crucial. A core concept in using PARP inhibitors (PARPi) for treating patients with BRCA1/2 mutations is the concept of synthetic lethality. Our research found that Rlip depletion or inhibition, combined with PARPi, works synergistically in both BRCA1/2-mutated and BRCA1/2-wild-type breast cancer cells. Methods: Two PARP inhibitors, AZD2281 (Olaparib) and AZD2461, were tested on the proliferation and survival of different cancer cell lines, including MDA-MB-231 (triple negative), MCF-7 (ER+), SKBR-3, AU565 (HER-positive), and MDA-MB-436 (BRCA1 mutated triple negative), with and without Rlip depletion using an antisense locked nucleic acid (LNA). For all experiments, 2.5 µM AZD2461 and 10 µM AZD2281 were used for 24 hours in complete growth medium. Cell cycle analysis was performed by flow cytometry. Expression levels of γH2AX and Rad51 proteins were assessed via Western blotting and immunofluorescence staining. The comet assay was used to measure DNA damage and fragmentation. Results: Rlip depletion significantly increased the reduction in cell survival caused by PARPi treatments across all tested cell lines. Combination index (CI) values were calculated using the Hill equation and Chou-Talalay’s method with CompuSyn software. The effects were highly synergistic (CI < 1) with both inhibitors in SK-BR-3 and MCF-7 cells. Olaparib showed additive effects in MDA-MB-231 cells, whereas AZD2461 exhibited an antagonistic effect. The combination of PARPi and Rlip depletion was antagonistic in AU565 and MDA-MB-436 (CI > 1), but the doses of both inhibitors were significantly reduced (p < 0.001) after Rlip depletion. Cell cycle analysis with Rlip depletion showed either an increase in the sub-G1 population (cell death) or G2/M phase arrest, suggesting cells attempt to undergo mitosis with unrepaired DNA damage, leading to elevated DNA double-strand break signaling, cell cycle arrest, and apoptosis. Alkaline comet assay confirmed more DNA damage with Rlip depletion compared to treatments with the inhibitor alone. Western blot analysis for Rad51 and γ-H2AX showed defects in protein levels following Rlip depletion. The number of γ-H2AX foci, characteristic of DNA damage in nuclei, increased significantly (p < 0.01) after Rlip depletion compared to inhibitor treatment alone. Conclusion: Our findings suggest that drugs that inhibit or deplete Rlip could help sensitize breast cancer cells, regardless of their BRCA status or whether they are wild-type ER+ or triple-negative breast cancers, to PARP inhibitors. The immunogenicity of HRD-associated malignancies and the immunomodulatory effects of PARP inhibition could be combined with Rlip inhibitors to treat HRD cancers. These findings clarify the underlying mechanisms and are expected to lead to the development of first-in-class therapies. Citation Format: C. Bose, S. Awasthi, T. Hutson, F. Sardela de Miranda, M. F. Mahecha, M. W. Melkus, R. Layeequr Rahman, S. P. Singh. Ralbp1 depletion downregulates DNA repair and potentiates PARP inhibitor activity in BRCA wild type and mutated breast cancer cell lines [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2025; 2025 Dec 9-12; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2026;32(4 Suppl):Abstract nr PS4-04-20.

ObjectiveNursing professionals are frequently exposed to a variety of chemical agents in healthcare settings, including disinfectants, anesthetic gases, cytotoxic drugs and other hazardous substances, raising concerns about potential genotoxic effects. This systematic review investigates the genotoxicity associated with occupational exposure commonly encountered by nursing staff.MethodsA systematic search conducted in August 2025 across PubMed, SCOPUS, and Web of Science identified 16 relevant studies that evaluated DNA damage using biomarkers, such as the micronucleus assay, comet assay, and chromosomal aberration tests in human or mammalian cells.ResultsAll included studies consistently reported genotoxic effects linked to occupational exposure, including increased frequencies of micronuclei, DNA strand breaks, and chromosomal alterations. Quality assessment indicated that 15 (out of 16) studies were of moderate to strong methodological rigor, supporting the reliability of these results.ConclusionThese findings underscore the need for enhanced monitoring and protective measures for nursing professionals at risk of long-term genetic damage. The micronucleus assay emerges as a suitable assay for biomonitoring these professionals.

Abstract Background: Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype associated with poor prognosis and high rates of metastasis, contributing to 15-24% of breast cancer-related deaths. The oncogene proline-, glutamic acid-, and leucine-rich protein 1 (PELP1) is frequently overexpressed in TNBC and is associated with poor survival. This study aimed to elucidate the critical role of PELP1 in DNA repair mechanisms of TNBC and to evaluate a novel therapeutic strategy using a PELP1 small-molecule inhibitor, SMIP34, in combination with topoisomerase inhibitors (TIs). Methods: PELP1-inducible knockdown (iKD) TNBC models (MDA-MB-231, BT-549, HCC-1806, SUM-149) were generated via lentiviral transduction using IPTG-inducible shRNA constructs. PELP1 signaling was inhibited pharmacologically using SMIP34. A drug screen of 140 FDA-approved compounds was performed using MTT assays to identify synergistic agents with SMIP34. Functional assays, including colony formation and Annexin V/PI apoptosis assays, were used to assess therapeutic efficacy. Mechanistic studies utilized RT-qPCR, Western blotting, comet assays, and DNA Damage Response Phosphorylation Arrays. Combination treatments were validated in cell line-derived xenografts , patient-derived organoids , and patient-derived xenografts . Results: Western blot analyses confirmed that IPTG induction resulted in a dose-dependent suppression of PELP1 expression in all four PELP1-iKD TNBC cell lines. PELP1 knockdown significantly impaired cell proliferation and colony formation while inducing apoptosis. In parallel, pharmacological inhibition using SMIP34 produced similar phenotypic outcomes, validating the functional relevance of PELP1 in TNBC cell survival. Importantly, PELP1 suppression led to a delayed DNA damage response (DDR), marked by elevated γ-H2AX and activation of key DDR kinases including p-ATM, p-ATR, and DNA-PKcs. SMIP34 treatment replicated these effects, confirming that both genetic and pharmacologic inhibition of PELP1 compromises DNA repair efficiency. A high-throughput MTT-based screen of 140 FDA-approved agents identified topoisomerase inhibitors (TIs) as potent synergistic agent with SMIP34. Co-treatment with SMIP34 and TIs resulted in significantly enhanced growth inhibition and apoptosis compared to monotherapy. This synergy was confirmed in PELP1-iKD cells, where knockdown further sensitized cells to TIs, providing genetic validation of PELP1’s role in TI resistance. Mechanistic studies showed that combination treatments (PELP1-iKD+TIs or SMIP34+TIs) markedly increased DNA damage and apoptotic signaling. Western blotting and phospho-antibody arrays revealed upregulation of DDR and checkpoint proteins, including γ-H2AX, p-CHK2, p-ATR, p-CHK1, p-BRCA1, and p21, indicating enhanced replication stress and cell cycle arrest. Comet assays demonstrated significantly greater DNA fragmentation with combination therapy compared to single agents. In preclinical models, SMIP34+TI treatment significantly suppressed organoid growth in patient-derived organoid cultures. In vivo, this combination led to marked tumor growth inhibition in both cell line-derived xenografts and patient-derived xenografts, outperforming either agent alone. No significant toxicity was observed, supporting the therapeutic potential of this approach. Conclusion: These findings highlight PELP1 as a critical modulator of DNA damage response in TNBC and support the development of combination strategies using SMIP34 and topoisomerase inhibitors. This approach offers a promising therapeutic avenue for improving outcomes in patients with TNBC. Citation Format: K. Nassar, J. Sanchez, D. Panneerdoss, E. Behnam, X. Yang, U. Pratap, M. Mahajan, S. Alejo, P. Subbarayalu, D. Zhou, G. Sareddy, M. Rao, S. Viswanadhapalli, R. K. Vadlamudi. Pelp1 inhibition disrupts dna repair and enhances topoisomerase inhibitor efficacy in triple negative breast cancer [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2025; 2025 Dec 9-12; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2026;32(4 Suppl):Abstract nr PS4-04-23.

Atrazine (ATZ), one of the most widely practiced herbicides, is recognized as an endocrine disruptor with insurmountable ecotoxicological consequences. While extensively studied in vertebrate models, its impact on early diverging metazoans remains poorly characterized. We investigated the effects of ATZ exposure on Hydra magnipapillata, considering its structural and functional alteration at organismal, cellular and molecular hierarchies. Feeding assays revealed dose- and duration-dependent morphological alterations, battery cell disruption and impairment of prey capture. Regeneration and budding were greatly inhibited, suggesting compromised stem cell activity. Acridine orange (AO) staining and comet assay confirmed apoptosis through DNA damage compounding with oxidative stress-mediated cytotoxicity. Transcription analysis revealed modulation of Hippo signalling (Hpo, LATS and Mob) and induction of oxidative stress markers (SOD, GST, CAT and Hsp70) together with upregulation of FoxO, reflecting conserved stress adaptive responses. Protein analysis revealed upregulation of StAR and downregulation of SRD5A2 and AKR1C2, demonstrating the disruption of steroidogenesis. These results indicate that ATZ interferes with conserved pathways associated with growth, stress resistance and steroid metabolism in Hydra, linking molecular alterations to impaired physiology. By integrating classical and molecular biomarkers, this study supports the use of Hydra as a sentinel model in ecotoxicology and provides comparative insights into evolutionary conservation of endocrine-disturbing effects of ATZ across metazoans.

This systematic review (SR) with meta-analysis investigates the genotoxicity potential of coke oven workers (COWs) exposed to polycyclic aromatic hydrocarbons (PAHs) through a comprehensive analysis of studies retrieved from PubMed, SCOPUS, and Web of Science. The comparisons were defined as standardized mean difference (SMD), and 95% confidence intervals (CIs) were established. A systematic search conducted in May 2025 identified 21 relevant studies, which employed different assays, such as the micronucleus assay (MA) and the comet assay (CA) in order to assess DNA damage. The outcomes suggested that 21 of the reviewed studies observed genotoxic effects related to this exposure, with 20 inducing micronucleus formation and chromosomal abnormalities. As for the quality assessment, a total of 18 studies were classified as Strong, and three (out of 21) were deemed as Moderate. No study was categorized as Weak, which proves our findings can be considered trustworthy. The meta-analysis (six studies) revealed a statistically significant difference between COWs and the control group, for both the MN (SMD = 0.70, 95% CI, 0.26-1.15, p = 0.002) and CA (SMD = 0.86, 95% CI, 0.34-1.38, p = 0.001) with high heterogeneity. We concluded that there is a potential for genotoxicity in COWs. This certainly shows the importance of further investigation and regulatory oversight to ensure coke oven professionals' safety. Also, we understand such findings are vital for clarifying the role of biomarkers related to genotoxicity due to this occupational exposure.

Therapeutic potential of 1,3,4-oxadiazole derivative in EAE-induced optic neuritis.

Cypermethrin and methyl parathion are both insecticides, though they belong to different chemical classes and have distinct characteristics and effects. Apart from these, their frequent use in agricultural and health sectors throughout West Bengal, these insecticides showed significant environmental rises, affecting different life forms. This present study firstly deals with different in vitro methods viz. MTT assay studies for the calculation of IC50 doses of cypermethrin and methyl parathion on human normal kidney cell line HEK-293, these results were further supported by DAPI staining and Comet assay study for direct effects of these two insecticides on cellular DNA and finally DCFDA staining as cellular stress induced ROS regeneration study. The IC50 value of cypermethrin was found to be 27 µM, while in case of methyl parathion it was found to be 14 µM. These in vitro study reports were finally concluded by in vivo (histological) studies to show the potential toxicity generating effects of these two frequently used chemicals on kidney sections of male albino mice (Mus musculus) . For in vivo experiments LD50 values of cypermethrin and methyl Parathion was calculted respectively along with total accumulation of these two insecticides that eventually leads to prominent histological changes viz. glomerular shrinkage, vacuole formation, fat depositions etc. was also studied. All of these experimental results pointed out, though both of the insecticides are capable for generating toxicity in cellular and tissue level, between them methyl parathion was found to be the highly toxic one.

Deciphering the Mechanisms of Cupferron Reproductive Toxicity: Insights from In Vitro Assays, Network Toxicology, and Molecular Docking.

This study investigated the individual and combined toxicological effects of lead (Pb) and endosulfan on Labeo rohita fingerlings following a 28-day sublethal exposure. Seven experimental groups were established, including control, low and high concentrations of Pb and endosulfan, and their respective combinations. Hematological, biochemical, neurotoxic, tissue bioaccumulation, and genotoxic biomarkers were assessed. Results revealed significant hematological disturbances characterized by reductions in red blood cell count, hemoglobin, and hematocrit, alongside elevated white blood cell counts. Biochemical analyses showed hyperglycemia, hypoproteinemia, dyslipidemia, increased creatinine and urea levels, and marked elevation of hepatic enzymes (ALT, AST, ALP). Neurotoxicity was evident through significant inhibition of acetylcholinesterase activity, with the greatest suppression observed under combined high-dose exposure. Tissue analysis demonstrated substantial accumulation of Pb and endosulfan in gill, liver, and kidney tissues, with co-exposure resulting in up to 1.7-fold higher accumulation than individual treatments. Genotoxicity assessment using the comet assay revealed pronounced DNA damage, with damaged erythrocytes exceeding 65% and a threefold increase in genetic damage index in the combined high-dose group. Two-way ANOVA confirmed significant Pb × endosulfan interactions, highlighting synergistic toxicity. Overall, the findings underscore the heightened ecological and food safety risks associated with concurrent heavy metal and pesticide contamination in freshwater ecosystems.