In vivo evaluation of the neurogenotoxic effects of exposure to validamycin A in neuroblasts of Drosophila melanogaster larval brain.

Agriculture commonly utilizes crop protection products to tackle infestations from fungi, parasites, insects, and weeds. Validamycin A, an inhibitor of trehalase, possesses antibiotic and antifungal attributes. Epidemiological evidence has led to concerns regarding a potential link between pesticide usage and neurodegenerative diseases. The fruit fly, Drosophila melanogaster, has been recognized as a reliable model for genetic research due to its significant genetic similarities with mammals. Here, we propose to use D. melanogaster as an effective in vivo model system to investigate the genotoxic risks associated with exposure to validamycin A. In this study, we performed a neurotoxic evaluation of validamycin A in D. melanogaster larvae. Several endpoints were evaluated, including toxicity, intracellular oxidative stress (reactive oxygen species), intestinal damage, larval behavior (crawling behavior, light/dark sensitivity assay, and temperature sensitivity assay), locomotor (climbing) behavior, and neurogenotoxic effects (impaired DNA via Comet assay, enhanced by Endo III and formamidopyrimidine DNA glycosylase [FPG]). The results showed that exposure to validamycin A, especially at higher doses (1 and 2.5 mM), induced DNA impairment in neuroblasts as observed by Comet assay. Both larvae and adults exhibited behavioral changes and produced reactive oxygen species. Most importantly, this research represents a pioneering effort to report neurogenotoxicity data specifically in Drosophila larval neuroblasts, thus underscoring the importance of this species as a testing model in exploring the biological impacts of validamycin A. The in vivo findings from the experiments are a valuable and novel addition to the existing validamycin A neurogenotoxicity database.

Drosophila melanogaster as a dynamic in vivo model organism reveals the hidden effects of interactions between microplastic/nanoplastic and heavy metals.

Plastic waste in different environments has been constantly transforming into microplastic/nanoplastic (MNPLs). As they may coexist with other contaminants, they may behave as vectors that transport various toxic trace elements, including metals. Because the impact of exposure to such matter on health still remains elusive, the abundant presence of MNPLs has lately become a pressing environmental issue. Researchers have been utilizing Drosophila melanogaster as a dynamic in vivo model in genetic research for some time. The fly has also recently gained wider recognition in toxicology and nanogenotoxicity studies. The use of nanoparticles in numerous medical and consumer products raises serious concern, since many in vitro studies have shown their toxic potential. However, there is rather limited in vivo research into nanomaterial genotoxicity using mice or other mammalians owing to high costs and ethical concerns. In this context, Drosophila, thanks to its genetic tractability, short life span, with its entire life cycle lasting about 10 days, and distinct developmental stages, renders this organism an excellent model in testing toxic effects mediated by MNPLs. This review therefore aims to encourage research entities to employ Drosophila as a model in their nanogenotoxicity experiments focusing on impact of MNPLs at the molecular level.

Genotoxicity mechanism of food preservative propionic acid in the in vivo Drosophila model: gut damage, oxidative stress, cellular immune response and DNA damage.

Propionic acid is a short-chain fatty acid that is the main fermentation product of the enteric microbiome. It is found naturally and added to foods as a preservative and evaluated by health authorities as safe for use in foods. However, propionic acid has been reported in the literature to be associated with both health and disease. The purpose of this work is to better understand how propionic acid affects Drosophila melanogaster by examining some of the effects of this compound on the D. melanogaster hemocytes. D. melanogaster was chosen as a suitable in vivo model to detect potential risks of propionic acid (at five concentrations ranging from 0.1 to 10 mM) used as a food preservative. Toxicity, cellular immune response, intracellular oxidative stress (reactive oxygen species, ROS), gut damage, and DNA damage (via Comet assay) were the end-points evaluated. Significant genotoxic effects were detected in selected cell targets in a concentration dependent manner, especially at two highest concentrations (5 and 10 mM) of propionic acid. This study is the first study reporting genotoxicity data in the hemocytes of Drosophila larvae, emphasizing the importance of D. melanogaster as a model organism in investigating the different biological effects caused by the ingested food preservative product.

Protective effects of resveratrol against genotoxicity induced by nano and bulk hydroxyapatite in Drosophila melanogaster.

Hydroxyapatite (HAp) is a naturally occurring calcium phosphate mineral predominantly used for its biocompatibility in a number of areas such as bone grafting, prosthesis coating in dentistry, and targeted drug delivery. Since the nano form of HAp (nHAp) has gained popularity attributed to a re-mineralizing effect in dental repair procedures, concerns have been raised over safety and biocompatibility of these nanoparticles (NP). This study, therefore, aimed to (1) investigate mechanisms of potential genotoxicity and enhanced generation of reactive oxygen species (ROS) initiated by bulk and nano forms of HAp and (2) test in vivo whether resveratrol, a type of natural phenol, might mitigate the extent of potential DNA damage. The size of nHAp was determined to be 192.13 ± 9.91 nm after dispersion using transmission electron microscopy (TEM). Drosophila melanogaster was employed as a model organism to determine the genotoxic potential and adverse effects of HAp by use of (comet assay), mutagenic and recombinogenic activity (wing spot test), and ROS-mediated damage. Drosophila wing-spot tests demonstrated that exposure to nontoxic bulk and nHAp concentrations (1, 2.5, 5 or 10 mM) produced no significant recombination effects or mutagenicity. However, bulk and nHAp at certain doses (2.5, 5 or 10 mM) induced genotoxicity in hemocytes and enhanced ROS production. Resveratrol was found to ameliorate the genotoxic effects induced by bulk HAp and nHAp in comet assay. Data demonstrate that treatment with nano and bulk Hap-induced DNA damage and increased ROS generation D. melanogaster which was alleviated by treatment with resveratrol.

In vivo effects of 1,4-dioxane on genotoxic parameters and behavioral alterations in Drosophila melanogaster.

1,4-Dioxane (DXN) is used as solvent in different consumer products including cosmetics, paints, surfactants, and waxes. In addition, DXN is released as an unwanted contaminating by-product as a result of some reactions including ethoxylation of alcohols, which occurs with in personal care products. Consequently, DXN pollution was detected in drinking water and is considered as an environmental problem. At present, the genotoxicity effects attributed to DXN are controversial. The present study using an in vivo model organism Drosophila melanogaster aimed to determine the toxic/genotoxic, mutagenic/recombinogenic, oxidative damage as evidenced by ROS production, phenotypic alterations as well as behavioral and developmental alterations that are closely related to neuronal functions. Data demonstrated that nontoxic DXN concentration (0.1, 0.25, 0.5, or 1%) induced mutagenic (1%) and recombinogenic (0.1, 0.25, or 0.5%) effects in wing spot test and genotoxicity in hemocytes using comet assay. The nontoxic concentrations of DXN (0.1, 0.25, 0.5, or 1%) significantly increased oxidative stress, climbing behavior, thermal sensivity and abnormal phenotypic alterations. Our findings show that in contrast to in vitro exposure, DXN using an in vivo model Drosophila melanogaster this compound exerts toxic and genotoxic effects. Data suggest that additional studies using other in vivo models are thus warranted.

Exposure to boron trioxide nanoparticles and ions cause oxidative stress, DNA damage, and phenotypic alterations in Drosophila melanogaster as an in vivo model.

Boron trioxide nanoparticles (B2 O3 NPs) have recently been widely used in a range of applications including electronic device technologies, acousto-optic apparatus fields, and as nanopowder for the production of special glasses. We propose Drosophila melanogaster as a useful in vivo model system to study the genotoxic risks associated with NP exposure. In this study, we have conducted a genotoxic evaluation of B2 O3 NPs (size average 55.52 ± 1.41 nm) and its ionic form in D. melanogaster. B2 O3 NPs were supplied to third instar larvae at concentrations ranging from 0.1-10 mM. Toxicity, intracellular oxidative stress (reactive oxygen species, ROS), phenotypic alterations, genotoxic effect (via the wing somatic mutation and recombination test, SMART), and DNA damage (via Comet assay) were the end-points evaluated. B2 O3 NPs did not cause any mutagenic/recombinogenic effects in all tested non-toxic concentrations in Drosophila SMART. Negative data were also obtained with the ionic form. Exposure to B2 O3 NPs and its ionic form (at two highest concentrations, 2.5 and 5 mM) was found to induce DNA damage in Comet assay. Additionally, ROS induction in hemocytes and phenotypic alterations were determined in the mouths and legs of Drosophila. This study is the first study reporting genotoxicity data in the somatic cells of Drosophila larvae, emphasizing the importance of D. melanogaster as a model organism in investigating the different biological effects in a concentration-dependent manner caused by B2 O3 NPs and its ionic form. The obtained in vivo results contribute to improvement the genotoxicity database on the B2 O3 NPs.

Hazard Assessment of the Effects of Acute and Chronic Exposure to Permethrin, Copper Hydroxide, Acephate, and Validamycin Nanopesticides on the Physiology of Drosophila: Novel Insights into the Cellular Internalization and Biological Effects.

New insights into the interactions between nanopesticides and edible plants are required in order to elucidate their impacts on human health and agriculture. Nanopesticides include formulations consisting of organic/inorganic nanoparticles. Drosophila melanogaster has become a powerful model in genetic research thanks to its genetic similarity to mammals. This project mainly aimed to generate new evidence for the toxic/genotoxic properties of different nanopesticides (a nanoemulsion (permethrin nanopesticides, 20 ± 5 nm), an inorganic nanoparticle as an active ingredient (copper(II) hydroxide [Cu(OH)2] nanopesticides, 15 ± 6 nm), a polymer-based nanopesticide (acephate nanopesticides, 55 ± 25 nm), and an inorganic nanoparticle associated with an organic active ingredient (validamycin nanopesticides, 1177 ± 220 nm)) and their microparticulate forms (i.e., permethrin, copper(II) sulfate pentahydrate (CuSO4·5H2O), acephate, and validamycin) widely used against agricultural pests, while also showing the merits of using Drosophila-a non-target in vivo eukaryotic model organism-in nanogenotoxicology studies. Significant biological effects were noted at the highest doses of permethrin (0.06 and 0.1 mM), permethrin nanopesticides (1 and 2.5 mM), CuSO4·5H2O (1 and 5 mM), acephate and acephate nanopesticides (1 and 5 mM, respectively), and validamycin and validamycin nanopesticides (1 and 2.5 mM, respectively). The results demonstrating the toxic/genotoxic potential of these nanopesticides through their impact on cellular internalization and gene expression represent significant contributions to future nanogenotoxicology studies.

Genotoxicity responses of single and mixed exposure to heavy metals (cadmium, silver, and copper) as environmental pollutants in Drosophila melanogaster.

Heavy metals are now persistently present in living things' environments, in addition to their potential toxicity. Therefore, the aim of this study was to utilize D. melanogaster to determine the biological effects induced by different heavy metals including cadmium chloride (CdCl2), copper (II) sulfate pentahydrate (CuSO 4.5 H2O), and silver nitrate (AgNO3). In vivo experiments were conducted utilizing three low and environmentally relevant concentrations from 0.01 to 0.5 mM under single and combined exposure scenarios on D. melanogaster larvae. The endpoints measured included viability, reactive oxygen species (ROS) generation and genotoxic effects using Comet assay and the wing-spot test. Results indicated that tested heavy metals were not toxic in the egg-to adult viability. However, combined exposure (CdCl2+AgNO3 and CdCl2+AgNO3+CuSO 4.5 H2O) resulted in significant genotoxic and unfavorable consequences, as well as antagonistic and/or synergistic effects on oxidative damage and genetic damage.

Photodynamic augmentation of oncolytic virus therapy for central nervous system malignancies.

Oncolytic viruses (OVs) have emerged as a clinical therapeutic modality potentially effective for cancers that evade conventional therapies, including central nervous system malignancies. Rationally designed combinatorial strategies can augment the efficacy of OVs by boosting tumor-selective cytotoxicity and modulating the tumor microenvironment (TME). Photodynamic therapy (PDT) of cancer not only mediates direct neoplastic cell death but also primes the TME to sensitize the tumor to secondary therapies, allowing for the combination of two potentially synergistic therapies with broader targets. Here, we created G47Δ-KR, clinical oncolytic herpes simplex virus G47Δ that expresses photosensitizer protein KillerRed (KR). Optical properties and cytotoxic effects of G47Δ-KR infection followed by amber LED illumination (peak wavelength: 585-595 nm) were examined in human glioblastoma (GBM) and malignant meningioma (MM) models in vitro. G47Δ-KR infection of tumor cells mediated KR expression that was activated by LED and produced reactive oxygen species, leading to cell death that was more robust than G47Δ-KR without light. In vivo, we tested photodynamic-oncolytic virus (PD-OV) therapy employing intratumoral injection of G47Δ-KR followed by laser light tumor irradiation (wavelength: 585 nm) in GBM and MM xenografts. PD-OV therapy was feasible in these models and resulted in potent anti-tumor effects that were superior to G47Δ-KR alone (without laser light) or laser light alone. RNA sequencing analysis of post-treatment tumor samples revealed PD-OV therapy-induced increases in TME infiltration of variable immune cell types. This study thus demonstrated the proof-of-concept that G47Δ-KR enables PD-OV therapy for neuro-oncological malignancies and warrants further research to advance potential clinical translation.

Interactions of Ingested Polystyrene Microplastics with Heavy Metals (Cadmium or Silver) as Environmental Pollutants: A Comprehensive In Vivo Study Using Drosophila melanogaster.

Living organisms are now constantly exposed to microplastics and nanoplastics (MNPLs), and besides their toxic potential, they can also act as carriers of various hazardous elements such as heavy metals. Therefore, this study explored possible interactions between polystyrene microplastics (PSMPLs) and two metal pollutants: cadmium chloride (CdCl2) and silver nitrate (AgNO3). To better understand the extent of biological effects caused by different sizes of PSMPLs, we conducted in vivo experiments with five doses (from 0.01 to 10 mM) that contained polystyrene particles measuring 4, 10, and 20 µm in size on Drosophila larvae. Additional experiments were performed by exposing larvae to two individual metals, CdCl2 (0.5 mM) and AgNO3 (0.5 mM), as well as combined exposure to PSMPLs (0.01 and 10 mM) and these metals, in an attempt to gain new insight into health risks of such co-exposure. Using transmission electron microscopy imaging, we managed to visualize the biodistribution of ingested PSMPLs throughout the fly's body, observing the interactions of such plastics with Drosophila intestinal lumen, cellular uptake by gut enterocytes, the passage of plastic particles through the intestinal barrier to leak into the hemolymph, and cellular uptake by hemocytes. Observations detected size and shape changes in the ingested PSMPLs. Egg-to-adult viability screening revealed no significant toxicity upon exposure to individual doses of tested materials; however, the combined exposure to plastic and metal particles induced aggravated genotoxic effects, including intestinal damage, genetic damage, and intracellular oxidative stress (ROS generation), with smaller sized plastic particles + metals (cadmium and silver) causing greater damage.

Heavy metal accumulation and genotoxic effects in levant vole (Microtus guentheri) collected from contaminated areas due to mining activities.

Heavy metal contamination is a serious environmental problem commonly monitored in various organisms. Small wild rodents are ideal biological monitors to show the extent of environmental pollution. The aim of this study was to evaluate the adverse effects of marble and stone quarries on the Levant vole, Microtus guentheri, inhabiting some polluted sites. In this context, Inductively Coupled Plasma Mass Spectrometry (ICP-MS) was used to analyze distribution of thirteen heavy metals (Fe, Al, Zn, Cu, Cr, Mn, Ni, B, Pb, As, Co, Cd, and Hg) in the organs (skins, bones, muscles, livers and kidneys) of the biological specimens, and the comet assay revealed DNA damage in blood lymphocytes for the first time. This study was conducted at close to the marble and stone quarries at Korkuteli, Antalya-Turkey during spring, summer, autumn (2017) and winter (2018) seasons. In spring and summer, genetic damage in blood lymphocytes from all polluted sites (sites 1-5) was significantly higher than that of controls, while in autumn it was higher in samples from three sites (sites 3-5). In terms of heavy metal distribution in organs, we found depositions of Fe, Al, Zn, Ni, Mn, Cr, Co, As and Pb primarily in the skin with its derivatives, Cu and Cd deposits in the kidney, Cu, Cd and B deposits in the liver, and As and Pb depositions in the bones. The study shows that certain organs (especially skin with its derivatives) and blood lymphocytes of Levant vole can be used as ideal indicators of heavy metal pollution. Our results suggest that the Korkuteli area could already be under the threat of heavy metal pollution.