Targeting DNA damage: A natural product-based strategy for inhibiting cancer progression.

Background: Infertility is a worldwide health problem that affects about 15-20% of couples globally. Various factors contribute to infertility, including physiological, genetic, hormonal factors, and environmental pollution. Cadmium chloride, a heavy metal pollutant, has an adverse impact on the reproductive system, while curcumin is recognized for its protective antioxidant properties. Objectives: The present study aimed to explore the protective effect of curcumin on sperm DNA integrity, hormonal parameters, and DNAH1 gene expression against the adverse effect of cadmium chloride exposure in male rats. Methods: Forty adult male Wistar rats were divided into four equal groups, including control (C), curcumin (CU), cadmium chloride (CD), and treatment (T) groups. The CU, CD, and T groups received curcumin via intraperitoneal injection. At the end of the experiment, samples of the epididymis, blood, and testes were collected to evaluate sperm DNA fragmentation, testosterone and luteinizing hormone (LH) levels, and DNAH1 gene expression. Results: Cadmium chloride significantly reduced testosterone levels from 0.1735±0.0082 ng/mL to 0.0986±0.0028 ng/mL, increased LH levels from 0.3907±0.0101 ng/mL to 0.5389±0.0384 ng/mL, and raised sperm DNA fragmentation from 5.63±0.34% to 20.66±0.58%, while DNAH1 gene expression dropped from 1.00±0.037 to 0.012±0.021. Curcumin restored testosterone levels to 0.1646±0.0076 ng/mL, reduced sperm DNA fragmentation to 3.93±0.44%, and increased DNAH1 expression levels to 2.69±0.061. Conclusion: Curcumin showed protective effects against the adverse and harmful effects of cadmium chloride that caused reproductive disorders and infertility by improving sperm DNA integrity, maintaining hormonal balance, and regulating gene expression.

Effects of different L-carnitine concentrations on post-thaw ram semen characteristics and fertilizing potential.

Analysis of urine cell-free DNA copy number and fragment size from healthy individuals.

Cryopreservation of rainbow trout (Oncorhynchus mykiss) sperm: Evaluation of cryomedia and protocol optimization.

Multimodal cell death induced by indirubin-3'-oxime through inhibition of Akt/mTOR axis in lung cancer cells.

Toward rapid testing for molecular authentication: Novel method for multianalyte identification of olive, sunflower, soy, sesame and corn DNA by visual biosensing.

Human biomonitoring studies aim to identify potential exposures to environmental, occupational, or lifestyle toxicants in human populations and are commonly used by public health decision-makers to predict disease risk. The Comet assay measures changes in genomic stability and is one of the most reliable biomarkers to indicate early biological effects, and therefore is accepted by various governmental regulatory agencies. The appeal of the Comet Assay lies in its relative simplicity, rapidity, sensitivity, and economic efficiency. Furthermore, the assay is known for its broad versatility, as it can be applied to virtually any human cell and easily adapted in order to detect particular biomarkers of interest, such as DNA repair capacity or single- and double-strand breaks. In a standard experiment, isolated single cells are first embedded in agarose and then lysed in high-salt solutions in order to remove all cellular contents except the DNA attached to a nuclear scaffold. Subsequent electrophoresis results in the accumulation of undamaged DNA sequences at the proximity of the nuclear scaffold, while damaged sequences migrate toward the anode. When visualized with fluorochromes, these migrated DNA fragments resemble a Comet tail and can be quantified for their intensity and shape according to internationally drafted guidelines.

Advancement of doxorubicin monitoring with a DNA fragmentation strategy for SYBR Green I-based aptamer biosensors.

Micro (nano)-plastics (MPs/NPs) are ubiquitously detected in human samples: stool, placenta, breastmilk, testes and semen, liver, lung, and blood, with their detailed toxicological profile still emerging. Numerous scientific studies are increasingly being conducted towards understanding possible deleterious effects of MPs/NPs on human, animal, plant, and environmental health. MPs/NPs rarely biodegrade, have small particle sizes, and are positively charged-features that make them dangerous to cells. They are capable of inducing oxidative stress, genotoxicity, immunological response, alteration in cellular membrane and cytoarchiture. The ability for MPs/NPs to induce DNA damage and genotoxicity may enhance genome instability, the hallmark of cancer and genetic disease syndrome. This review focused on determining the sensitivity of various biomarkers utilized to assess the DNA damage and genotoxicity potentials of MPs/NPs, the bioindicators used as models (invertebrates, vertebrates, cell lines/primary cells and plants), and the mechanisms of MPs/NPs-induced genotoxicity and DNA damage. The nature/type and size of the MP/NP polymers, concentration, and duration of exposure are determining factors considered in the DNA damage and genotoxicity assessment of MPs/NPs. Also, somatic and germ-line cells are susceptible to the genotoxic effects of MPs/NPs. Single and double DNA strand breaks assessed using the comet assay are the most used biomarker of DNA damage, while chromosome aberration is the least used. Others are sperm morphology assay, micronucleus assay, and toxicogenomics. The mechanisms of MPs/NPs-induced DNA damage include generation of free radicals and oxidative stress, induction of deleterious inflammatory cells, down-regulation of transcriptional genes related to apoptotic expressions, and increased DNA fragmentation in cells and tissues. Further studies are required to unequivocally confirm MPs/NPs as genotoxins, mutagens, and/or carcinogens.

ABSTRACT Cryopreservation of sperm is routinely used in assisted reproduction technology (ART) for male fertility preservation. However, this method has been associated with oxidative stress and DNA fragmentation that may impair sperm quality. Additionally, antioxidant interventions such as melatonin supplementation have not been thoroughly explored in this setting. Although Libya is reported to have one of the highest global prevalence rates of male infertility, Libya-specific data remain limited. This study aimed to determine the effect of a single freeze–thaw cycle on sperm DNA fragmentation and oxidative stress markers, and to evaluate whether melatonin has an impact on post-thaw oxidation profiles. This prospective cohort study was conducted at the Fertility and Reproductive Medicine Center, Beirut Hospital, Benghazi. Semen samples of 104 normozoospermic Libyans were evaluated before and after freezing. DNA fragmentation index (DFI) was measured by sperm chromatin dispersion (SCD) test, and reactive oxygen species (ROS) were quantified by using luminol-enhanced chemiluminescence. In a subset of ejaculates, aliquots were supplemented with 2 mM of melatonin prior to cryopreservation. Cryopreservation was associated with a statistically significant increase in DFI (46.3 ± 18.3% to 60.0 ± 23.0%; p < 0.001) and ROS levels (3.2 × 10³ to 14.7 × 10³ RLU/s; p < 0.001). Smokers presented significantly higher DFI at both pre-freeze and post-thaw evaluations (p < 0.001). We detected a positive correlation between ROS and post-thaw DFI (r = 0.68; p < 0.001). Melatonin-treated samples exhibited moderate but significant differences in ROS (12%, p = 0.045) and DFI (11%, p = 0.004) compared to untreated aliquots. These findings suggested that the freeze–thaw process may contribute to oxidative and genomic stress in spermatozoa, while melatonin supplementation appears to provide limited protection. Larger, multicenter studies incorporating ART endpoints are required to determine the potential translational relevance of these findings.

ABSTRACT Ovarian cancer ranks as the sixth most prevalent type of gynecological cancer. Smp43, a cationic antimicrobial peptide isolated from the scorpion venom of Scorpio maurus palmatus, exhibits notable antibacterial, against both Gram-positive and Gram-negative bacteria, and antifungal activities. This study evaluates the anti-cancer efficacy of Smp43 and examines its effects on cell viability, cell cycle progression, apoptosis, necrosis, and oxidative stress in a human ovarian cancer cell line (OVCAR-3). Smp43 significantly reduced the viability of OVCAR-3 cells compared to the normal fibroblast cell line WI-38, with IC50 values of 7.75 µg/mL and 29.50 µg/mL, respectively. The peptide effectively caused G1 phase cell cycle arrest and apoptosis in OVCAR-3 cells. It modulated apoptotic markers by downregulating the pro-survival marker Bcl-2 while upregulating the pro-apoptotic markers Bax, p53, caspase-3, caspase-8, and caspase-9. Additionally, Smp43 significantly increased DNA fragmentation in OVCAR-3 cells and decreased antioxidant parameters. These findings suggest that Smp43 possesses potential anti-ovarian carcinoma properties, exerting its effects through mechanisms involving apoptosis induction, necrosis, G1 cell cycle arrest, and inhibition of the antioxidant defense system.

DNA methyltransferases (MTases) play a critical role in epigenetic regulation and are closely associated with the occurrence and development of various diseases. To address the limitations of current detection methods, which often suffer from complicated operation and limited sensitivity we have developed a label-free fluorescent biosensing platform based on DNA-templated silver nanoclusters (DNA-AgNCs) for the detection of DNA adenine methyltransferase (Dam MTase) activity and the screening of its inhibitors. In this system, DNA-AgNCs signal probes are synthesized via a simple reduction reaction. Dumbbell-shaped DNA substrates are designed to be specifically methylated by Dam MTase in the presence of S-adenosylmethionine (SAM). The methylated substrates are subsequently recognized and cleaved by DpnI, yielding DNA fragments with 3'-OH termini. These termini are extended by terminal deoxynucleotidyl transferase (TdT) in the presence of dGTP, generating G-rich sequences, which hybridize with the DNA-AgNCs and significantly enhance the fluorescence intensity. This cascade reaction enables the sensitive detection of Dam MTase activity. Compared with the other approaches, the proposed method exhibits a wider linear range (0.5-10 U/mL) and a lower detection limit (0.165 U/mL). Furthermore, inhibition studies demonstrate that 5-fluorouracil and gentamicin effectively suppress Dam MTase activity. The method also shows excellent performance in complex biological samples. Overall, this fluorescence-based strategy offers high sensitivity and excellent specificity, demonstrating broad application prospects in epigenetic research and drug discovery.

Pharyngeal squamous cell carcinoma (PSCC) is an aggressive subtype of head and neck squamous cell carcinoma (HNSCC) with poor prognosis and low survival rates. Immune checkpoint inhibitors (ICIs) show promise, but less than 20% of HNSCC patients respond positively. Targeted radionuclide therapy (TRT) combines radionuclides with monoclonal antibodies to target tumor cells. This study created a reliable animal model of PSCC for evaluating the therapeutic efficacy of 131I-aPD-L1. Nude mice were subcutaneously implanted with FaDu cells-a human PSCC cell line characterized by high PD-L1 expression. The synthesis of 131I-aPD-L1 was optimized by varying labeling conditions, achieving a labeling efficiency of over 90%. Mice were divided into experimental and control groups; the experimental group received a single intravenous injection of 500 μCi 131I-aPD-L1. Accumulation of 131I-aPD-L1 in tumor tissues was confirmed by animal single-photon emission computed tomography (SPECT). Tumor volume and mouse body weight were measured every 3 days for 30 days. At the end of the study, tumor tissues were stained for histological examination and immunohistochemical analysis of Bcl-2 and Caspase-3 expression levels. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was also performed on tumor tissues. SPECT verified a significant accumulation of 131I in FaDu tumor tissue. The experimental group exhibited significantly slower tumor volume increase compared to the control group (t = 2.37, p < 0.05). Additionally, a significant reduction in body weight was observed in the 131I-aPD-L1 group compared to the control group (t = 5.624, p < 0.01). HE staining showed extensive tumor necrosis in the experimental group. Immunohistochemical analysis revealed negative Bcl-2 expression and higher caspase-3 expression in the experimental group, indicating enhanced apoptosis and necrosis in tumor cells. Furthermore, TUNEL assay further confirmed that 131I exerted cytotoxic effects by inducing DNA fragmentation. Collectively, our findings demonstrate the promising therapeutic potential of 131I-aPD-L1 for PSCC, particularly in patients with drug resistance or recurrent head and neck tumors. However, the use of nude mice may have impacted the full therapeutic efficacy and synergistic potential observed with immunotherapy. Future studies should utilize immunocompetent models to better assess the probe's therapeutic impact and to explore its synergistic effects with immunotherapy and reduce the dose of 131I to mitigate its toxic effects.

ABSRACT Introduction. In recent years, increasing attention has been paid to the issue of male reproductive health. The WHO guidelines state that the primary assessment of male fertility is reduced to a “basic semen examination” (concentration, morphology assessment, etc. of sperm). The study of DNA fragmentation, on the other hand, falls under the “expanded” list of tests. At the same time, a thorough evaluation of the antioxidant system’s activity and, in particular, the amount of oxidative modification products of macromolecules, is not included in this list. Sperm are sensitive cells to the effects of reactive oxygen species. Current research focuses on their condition and microenvironment, aimed at detecting the relationship between sperm quality and the development of oxidative stress. However, the question of whether to conduct an expanded and in-depth analysis in cases of normal sperm motility remains unresolved. Objective. The aim of this study was to evaluate the parameters of the antioxidant system and products of oxidative modifications of proteins and DNA of seminal plasma and spermatozoa at a normal level of their mobility and varying DNA fragmentation. Material and methods. This selective pilot single-center cross-sectional study included healthy men with different DNA fragmentation and normal sperm motility. We assessed the oxidative modifications of macromolecules (8-hydroxy-2’-deoxyguanosine; nitrotyrosine) and components of the antioxidant system (total antioxidant activity, catalase activity, superoxide dismutase activity, uric acid, and zinc ions) in seminal plasma and the cellular fraction of the ejaculate. Differences were considered significant at p&lt;0.050. Results. Using the exclusion criteria, we selected 37 patients. In the seminal plasma of men with TUNEL&gt;15, a decrease in superoxide dismutase activity (p&lt;0.010), zinc ion levels (p&lt;0.050) and uric acid content (p&lt;0.050) was observed, while an increase in nitrotyrosine content (p&lt;0.050) was observed only in the cellular fraction of the ejaculate. Conclusions. The obtained data indicate that in men, despite normal sperm motility, a high degree of DNA fragmentation is associated with abnormal head morphology, highlighting the necessity of conducting an “expanded” semen examination. Changes in the components of the antioxidant system are found not within the cells themselves but in their microenvironment, which may likely lead to an increased formation of oxidative modification products of macromolecules in sperm. This is important to consider when collecting samples for ART protocols, as well as in relation to the overall quality and reproductive potential of sperm, which is crucial when planning for pregnancy.

ObjectiveTo analyze the incidence of different types of Y chromosome microdeletions in infertile male patients in China, and to investigate the relationship between microdeletions in different azoospermia factor (AZF) regions and sperm kinetic parameters, sperm morphological parameters, and sex hormone levels.MethodsA total of 2010 infertile male patients who visited the Fujian Provincial Maternity and Child Health Hospital from 2022 to 2025 were selected. Their Y chromosome microdeletions (YCMD), semen routine, sperm morphology, sperm DNA fragmentation index (DFI), and sex hormone levels were detected, and the relationships between these parameters were analyzed.ResultsThe incidence of Y chromosome microdeletions in patients was 8.66% (174/2010). Among the 174 patients with AZF microdeletions, the proportion of AZFc region deletions was 85.63% (149/174), AZFa region deletions accounted for 2.30% (4/174), AZFb/c region deletions accounted for 8.05% (14/174), AZFa/b/c region deletions accounted for 2.87% (5/174), and heterochromosome deletions accounted for 1.15% (2/174). There were no statistically significant differences in semen volume, testosterone (T), and prolactin (PRL) levels between patients with different types of AZF deletions and the normal group (P>0.05). There were statistically significant differences in sperm concentration, progressive motility (PR), non-progressive motility (NP), total sperm motility, normal sperm morphology rate, sperm DFI, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) between patients with different types of AZF deletions and the normal group (P<0.05).ConclusionAZFc deletion is the most common type of Y chromosome microdeletion in infertile male patients in China. Patients with AZFa and AZFa/b/c combined deletions often present with azoospermia. AZFc deletion is associated with abnormal sperm quality parameters and disordered hormone levels.

Transposable elements (TEs), commonly referred to as “jumping genes,” are discrete DNA fragments capable of mobilization within the genome, often creating new copies during transposition. Once considered non-functional “junk DNA,” TEs are now recognized as essential regulators of genome evolution, gene networks, and epigenetic processes. They contribute to genome innovation through gene duplication, exon shuffling, regulatory rewiring, and stress-responsive plasticity. Conversely, their uncontrolled activation can disrupt genetic integrity. Such aberrant TE activity has been associated with immune dysfunction, cancer, neurodegeneration, and reproductive disorders. In livestock, recent studies have identified TE-derived regulatory elements that influence growth, reproduction, immunity, and adaptation. Notably, TEs constitute approximately 45–60% of the genomes of major livestock species, including pigs, sheep, and cattle, highlighting their extensive regulatory impact. Advances in genome editing and artificial intelligence have facilitated their functional characterization. This allows a sharper focus on TE-driven regulation in animal genomes without repeatedly specifying the livestock context. Emerging technologies such as CRISPR–transposon systems, programmable epigenome editing, and AI-driven TE mapping offer new opportunities for precise genome engineering, functional genomics, and livestock breeding. This review consolidates current knowledge on TE classification, molecular mechanisms, evolutionary roles, disease associations, and biotechnological applications, with emphasis on livestock genomics. It also outlines existing challenges, biosafety considerations, and future directions toward harnessing TEs for precision animal breeding and synthetic biology.

Objectives In the investigations of infertile couples, the contribution of male partners has recently caught the attention of researchers, and more and more investigations like DNA fragmentation tests, microfluidics, physiological intracytoplasmic sperm injection, and intracytoplasmic morphological sperm injection are being resorted to for better Assisted Reproductive Technology (ART) results. Male fertility is intrinsically linked to overall health, with a growing body of evidence indicating that medical comorbidities and conditions detrimental to men’s health are consistently associated with compromised reproductive function. Considering the fact that 15% of the male human genome is dedicated to reproductive functions, it is plausible that other health disorders may also be associated with impairments in fertility. This study was planned to look into factors which are causing such a rise in male infertility and its association with various semen parameters. Material and Methods A cross-sectional study was undertaken over an 18-month period at the infertility clinic of a tertiary care centre, enrolling 151 infertile males exhibiting abnormal semen parameters in accordance with the WHO 2010 guidelines. A comprehensive evaluation of their biochemical and endocrinological profiles was performed, and the correlation between these parameters and semen abnormalities was systematically examined. Results A significant negative correlation was observed between various semen parameters and diastolic blood pressure (DBP), prolactin, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), follicle-stimulating hormone (FSH), and oestrogen. DBP (mmHg) correlated negatively with sperm concentration (million/ml) (R = −0.161). CRP (mg/l) correlated with sperm concentration and total sperm count (×10 6 /ejaculate) (R = −0.180 and −0.208, respectively). ESR (mm/hour) correlated with sperm concentration and total sperm count (R = −0.214 and −0.198, respectively). FSH (IU/l) correlated with sperm concentration and total sperm count (R = −0.216 and −0.206, respectively). Prolactin (μg/l) correlated with sperm concentration, total sperm count, and total motile sperm count (TMSC) (R = −0.210, −0.264, and −0.191, respectively). Oestrogen (pg/ml) showed the strongest negative correlation with sperm concentration, total sperm count, and TMSC (R = −0.387, −0.357, and −0.171, respectively). Conversely, significant positive correlations were observed between semen parameters and both uric acid and lipid profile. Serum uric acid (mg/dl) correlated positively with sperm morphology (%) (R = 0.203). Low-density lipoprotein (mg/dl) correlated with sperm concentration and total sperm count (R = 0.231 and 0.259, respectively), while triglycerides (mg/dl) correlated with sperm concentration and total sperm count (R = 0.197 and 0.204, respectively). However, triglycerides also showed a significant negative correlation with total motility (%) and progressive motility (%) (R = −0.186 and −0.180, respectively). Conclusion Our findings demonstrated that DBP, prolactin, ESR, CRP, FSH, and oestrogen exhibited significant negative correlations with various semen parameters, whereas uric acid and lipid profile parameters showed significant positive correlations. These results suggest that systemic health factors exert a considerable influence on male reproductive potential, highlighting the importance of evaluating overall health status in the assessment and management of male infertility.

DNA damage at the level of individual cells is assessed using the method of comet assay. This method allows detection of DNA damage caused by various factors, such as carcinogens, radiation and oxidative stress. In this study, we evaluated the genetic activity of ethanol extracts of four representatives of the Sal via genus using D. melanogaster as a model object. Larvae of the Canton-S laboratory strain were cultured on substrates with the addition of sage extracts ( S. stepposa Des-Shost., S. verticillata L., S. tesquicola Klokov and Pobed., S. glutinosa L) under their concentration in the medium of 0.1% and 1%. According to the results obtained, all the studied sage species are not characterized by genotoxicity at a concentration of 0.1%. Under this concentration, S. glutinosa demonstrates antigenotoxic properties, reducing the level of DNA fragmentation relative to the control. When increasing the concentration of the extract in the nutrient substrate to 1%, two representatives of the Salvia genus – S. stepposa and S. verticillate – become to manifest genotoxic properties. Thus, the established concentrations of plant extracts of Salvia species are promising for further study in order to identify effective biological properties.

This study aims to elucidate the physiological and biochemical alterations induced by parasitic Cuscuta sp. (dodder) in lucerne (Medicago sativa L.), a key forage crop. Comparative analyses between infected and healthy plants revealed that significant reductions in chlorophyll a, b, and total chlorophyll, and protein levels in the leaf and stem tissues of Cuscuta-infested plants were evident. The parasitic infection led to increased activities in antioxidant enzymes such as catalase (CAT) and peroxidase (POX) in stems, but not in leaves. Phenolic compounds were significantly lower both in leaves and stems of dodder-infected lucerne plants. No statistically significant changes were detected in jasmonic acid (JA) and salicylic acid (SA) levels in both plant parts, suggesting that classical defense signaling pathways may not be predominantly activated under Cuscuta-mediated stress. Possibly, host defense might be impaired. Histological examinations demonstrated active structural defense responses, including localized tissue remodeling and the formation of callose-like structures at haustorial penetration sites. DNA fragmentations showed that Cuscuta-infected M. sativa plants exhibited slightly higher instability. Collectively, these findings provide novel insights into the molecular and biochemical basis of the Cuscuta-lucerne interactions and highlight the need for further investigation into host defense mechanisms. We assume that active defense structural parts at early growth stages of lucerne or hypersensitive-type responses occurring in the early penetration phase might fend off the invading holoparasite. The results also offer a valuable foundation for the development of Cuscuta-resistant lucerne cultivars and support the design of integrated, sustainable weed management strategies to mitigate the detrimental effects of parasitic plants on forage production systems.