Search Results (71)

Background and Objectives: Transfusion-related immunomodulation (TRIM) is a recognized immunosuppressive phenomenon whose interaction with immune checkpoint blockade remains poorly defined. We evaluated the impact of erythrocyte suspension transfusion on survival and immune-related toxicity in patients with metastatic non-small cell lung cancer (NSCLC) treated with nivolumab. Materials and Methods: This retrospective, multicenter study included 253 patients with metastatic NSCLC treated with nivolumab across five centers between April 2018 and March 2025. Patients who received a transfusion within three months before or during nivolumab therapy were compared with non-transfused patients. Survival was assessed using Kaplan–Meier and Cox regression analyses. Results: Forty-two patients (16.6%) received transfusion. Transfused patients had significantly shorter progression-free survival (PFS) (median 3.6 vs. 9.3 months; HR = 2.28, 95% CI 1.47–3.56, p < 0.001) and overall survival (OS) (median 5.8 vs. 16.3 months; HR = 2.76, 95% CI 1.86–4.11, p < 0.001). In multivariable analysis, transfusion was an independent predictor of both shorter PFS (HR = 2.288, 95% CI 1.470–3.561, p < 0.001) and shorter OS (HR = 2.351, 95% CI 1.565–3.533, p < 0.001), independent of transfusion volume. No statistically significant association was detected between transfusion status and the incidence of nivolumab-related toxicity (p = 0.585), although this analysis was likely underpowered, whereas nivolumab-related toxicity was an independent favorable prognostic factor. Conclusions: Erythrocyte transfusion independently predicted poorer survival but not immune-related toxicity in nivolumab-treated metastatic NSCLC, supporting more judicious transfusion practices during immunotherapy. Full article

Acinetobacter baumannii is a microorganism of major global clinical importance, whose carbapenem-resistant phenotype is considered a priority target for the development of new antimicrobial agents. Carbapenem-resistant A. baumannii is associated with healthcare-associated infections and exhibits high mortality rates, prolonged hospital stays, and extensive use of intensive care units. As a control strategy, green-synthesized silver nanoparticles have gained increasing attention. Based on this, the aim of this study was to synthesize a novel triphasic silver nanocomposite, characterize it, and evaluate its antibacterial activity against extensively drug-resistant and pan-drug-resistant isolates. The nanocomposite was synthesized using cell-free bacterial supernatant and was physicochemically characterized regarding concentration, size, morphology, and crystallinity. Its antibacterial activity was assessed using broth microdilution and time–kill assays, followed by toxicity assays using human erythrocytes. A nanoscale and stable nanocomposite was obtained and was composed of metallic silver, silver chloride, and silver phosphate, which exhibited antibacterial activity against all tested isolates. In the toxicity assay, the nanocomposite demonstrated low toxicity and a high selectivity index. These findings demonstrate that Ag/AgCl/Ag₃PO₄ nanocomposite exhibits strong activity against resistant phenotypes for which therapeutic options are limited, highlighting its potential as a control strategy against this pathogen. Full article

In recent decades, intensified human activities, including industrialization and urban expansion, have given rise to significant environmental challenges, with plastic pollution emerging as a critical concern. Microplastics, particularly those derived from polystyrene (PS-MPs), are widespread in soils and oceans, posing risks to biodiversity and human health through ingestion, inhalation, and contact. This study examined the detrimental impact of PS-MPs on Zebrafish-Danio rerio (Hamilton, 1822) erythrocytes. The zebrafish (Danio rerio) is a well-established model organism in biomedical research, whereas its erythrocytes served as the cellular model investigated in this study. In vitro experiments involved exposing zebrafish erythrocytes, isolated from 25 adult zebrafish, to 105 µg/mL of PS-MPs for 30, 60 and 90 min to assess cytotoxicity, genotoxicity and oxidative stress. The results revealed a time-dependent and statistically significant reduction in erythrocyte viability (−30% to −50%), genomic stability (%GTS −20% to −40%), and a concomitant increase in DNA damage (DFI ~30–38%) and ROS production (~27–35%) compared to controls (all p ≤ 0.05). These molecular disruptions related to reduced cell viability, indicating that PS-MPs are cytotoxic and genotoxic, causing genetic damage and oxidative imbalance. The results obtained confirmed the toxicity of PS-MPs. These data suggest that PS-MPs can interfere with genetic material by inducing apoptosis through oxidative imbalance. Full article

The rapid emergence of antimicrobial resistance has intensified the search for alternative antimicrobial scaffolds that target both planktonic bacteria and biofilm-associated infections. In this study, a series of hydroxylated phosphonium ionic liquids derived from triphenylphosphonium (TPP⁺) and trihexylphosphonium (THP⁺) cations bearing C3, C6, C7, and C10 ω-hydroxyalkyl chains were synthesized and evaluated for their antibacterial and anti-biofilm activities. Antibacterial activity was determined using broth microdilution assays against Staphylococcus aureus and Escherichia coli, while anti-biofilm activity was assessed by disrupting preformed biofilms using a 96-pin microtiter plate system and crystal violet staining. The results showed that antibacterial activity was strongly influenced by the amphiphilic balance of the compounds, particularly the alkyl chain length and the nature of the phosphonium core. Derivatives bearing C6OH–C10OH chains exhibited the highest antibacterial activity, whereas short-chain analogs displayed markedly reduced potency. THP derivatives were notably more active against E. coli bacteria, consistent with their higher hydrophobicity and activity consistent with membrane interaction. In addition, THP derivatives demonstrated greater biofilm disruption, achieving up to ~90% biomass removal in E. coli biofilms, with C6OH–C7OH derivatives showing the most favorable activity profile. Hemolysis assays indicated low erythrocyte toxicity at concentrations close to antibacterial MIC values, indicating a favorable selectivity window. Overall, these findings highlight phosphonium ionic liquids as promising antimicrobial agents with activity consistent with membrane interaction and provide structure–activity insights for the rational design of new antibacterial and anti-biofilm compounds. Full article

Aspergillus cristatus is a key microorganism involved in the fermentation of Fu brick tea, but systematic strain-level safety data remain limited despite its long history of use in dark tea production. In this study, the toxicological safety of A. cristatus CCNH008 was assessed through acute oral toxicity assay, a 90-day repeated oral toxicity study with a recovery period and genotoxicity assays. In acute oral toxicity tests, CCNH008 caused no mortality, no adverse clinical signs, and no treatment-related effects on bodyweight or pathology in mice or rats, with an LD₅₀ greater than 10 g/kg bodyweight. In the 90-day repeated oral toxicity study in rats, the oral administration of CCNH008 at doses up to 1.67 g/kg bodyweight/day produced no treatment-related changes in clinical signs, bodyweight, food consumption, hematological or biochemical parameters, urinalysis, organ weights, or histopathology, including during a 28-day recovery period. Genotoxicity was evaluated using a bacterial reverse mutation assay, an in vitro mammalian chromosome aberration test, and an in vivo mammalian erythrocyte micronucleus test; no mutagenic or clastogenic effects were observed. Collectively, these findings demonstrate that CCNH008 shows no evidence of acute, subchronic oral toxicity, or genotoxicity, providing important strain-level safety evidence to support its potential application in food-related products. Full article

The potential effects of microplastics (MPs) on humans and ecosystems are of great concern, and it has been reported that the ingestion of contaminated food is the main route of exposure. In the present study, Perca fluviatilis was selected as a vertebrate model to evaluate the possible cellular effects induced by five different plastic polymers and car tire debris (CT) after 4- and 7-month exposure periods. The Cytome assay was chosen to check chromatin alteration in perch’s peripheral blood. The results indicated an increase in micronuclei and cytotoxic effect in specimens co-exposed to MPs + CT for 7 months. Increases in dicentric chromosomes were observed in specimens exposed both to MPs alone and to the mixture of MPs + CT, indicating for the first time a genotoxic effect induced by CT debris in fish in terms of structural aberrations. Increases in micronucleated erythrocyte frequency assessed after 7 months only after the addition of CT debris to the mixture of MPs might suggest an aneugenic action of CT in fish. In the same groups, the higher values of frequency in 8-shaped erythrocytes also indicate possible cell cycle toxicity exerted by CT exposure. An association between total erythrocyte nuclear morphology abnormalities (ENA) and glutathione reductase activity was also found, indicating a potential involvement of oxidative processes in modulating the genotoxicity observed. The present experimental model is a useful tool to study cellular mechanisms related to both MP- and CT-induced chromatin structure alterations indicating possible interference with human health as well. Full article

Methanolic bee pollen extracts from Xyris spp. and Mimosa spp. were evaluated for proximate composition, extraction yield, total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity, erythrocyte oxidative hemolysis protection, HRBC membrane stabilization, and LC-QToF-MS-based phytochemical annotation. Mimosa spp. pollen showed higher crude protein content than Xyris spp. pollen (29.54% vs. 18.38%, dry basis), whereas Xyris spp. pollen showed higher crude fat and nitrogen-free extract. The methanolic extraction yield was higher for Xyris spp. than Mimosa spp. (44.26% vs. 34.63%). Mimosa spp. extract exhibited higher TPC (17.07 ± 0.19 mg GAE/g dry pollen) and TFC (8.89 ± 0.59 mg QE/g dry pollen) than Xyris spp. extract, which contained 10.39 ± 0.27 mg GAE/g dry pollen and 6.94 ± 0.22 mg QE/g dry pollen, respectively. Mimosa spp. also showed lower DPPH and ABTS IC₅₀ values and higher FRAP activity. LC-QToF-MS results were reported as putatively annotated compounds based on accurate mass and database matching. These findings suggest that botanical origin influences the chemical composition and in vitro bioactivity of bee pollen; however, further targeted compound confirmation, toxicity assessment, and in vivo studies are required. Full article

Background/Objectives: The aquaculture industry represents a fundamental food sector. One of the main limiting factors for this sector is related to bacterial diseases, for which antibiotics have been widely used worldwide for decades. In recent years, a more conscious approach to the use of antimicrobials within the framework of the One Health approach has increased the need for alternatives capable of helping with disease management while avoiding the onset of antimicrobial resistance phenomena. Antimicrobial peptides, which have a broad spectrum of action against pathogens, are a promising solution. Methods: In this work, we investigated the capability of three peptides (Trematocine, Chionodracine, and Cnd-m3) isolated from Antarctic fish to target bacterial and viral pathogens affecting aquaculture. Successively, we investigated their cytotoxicity versus a continuous embryonic cell line (DLEC) derived from European sea bass and their haemolytic activity against fish erythrocytes. Moreover, we evaluated their immunomodulatory effect. Results: Regarding antibacterial properties, Cnd-m3 was identified as the best peptide, demonstrating good bactericidal and bacteriostatic activity against various bacterial strains, including Lactococcus garvieae. Concerning this bacterium, ANS permeability assays showed that the Cnd-m3 peptide has a great ability to interact with its outer membrane, while TEM analysis revealed that the peptide, after destabilization of the cell membrane, interacts with nucleic structures. Considering the antiviral activity, Trematocine was effective against two tested pathogenic enveloped viruses. Moreover, the toxicity of Trematocine and Cnd-m3 was evaluated by investigating their cytotoxicity against a cell line derived from Dicentrarchus labrax and haemolysis against sea bass erythrocytes. Both revealed good selectivity towards pathogens at the lowest concentration. Finally, Cnd-m3 manifested light in vitro immunomodulatory properties. Conclusions: Overall, these data provide a solid basis for future studies assessing the potential applications of two of the tested peptides in aquaculture. Full article

The growing global protein demand and environmental concerns from conventional animal agriculture have driven the exploration of sustainable alternative protein sources. Single-cell proteins (SCPs) from microbial fermentation offer a promising solution. This study comprehensively evaluated the nutritional value and safety profile of SCP produced from Ralstonia eutropha H16 through integrated in vitro and in vivo assessments. Nutritional analyses revealed a high crude protein content of 71.87 ± 5.05 g/100 g dry weight, with total amino acids of 53.67 ± 1.05 g/100 g. The essential amino acid content was 24.38 ± 0.51 g/100 g, accounting for 45% of the total amino acids. An essential amino acid index (EAAI) of 1.46 ± 0.04 and an amino acid score (AAS) of 0.83 ± 0.06 confirmed its classification as a high-quality protein source according to FAO/WHO standards. In vivo rat feeding trials demonstrated an adjusted protein efficiency ratio (PER) of 1.81, exceeding common plant proteins such as wheat (0.8–1.1). True digestibility (TD) reached 85.73%, with a biological value (BV) of 49.37%, net protein utilization (NPU) of 42.33%, and protein digestibility-corrected amino acid score (PDCAAS) of 0.71. Comprehensive safety assessments included chemical contaminant screening, acute oral toxicity studies in rats and mice, in vitro chromosome aberration tests, and erythrocyte micronucleus tests. Heavy metals and aflatoxin B₁ levels were below regulatory limits. Acute oral toxicity studies established LD₅₀ values exceeding 10,000 mg/kg body weight in both rodent species, classifying this protein source as practically non-toxic. The 28-day sub-acute toxicity study showed no significant adverse effects at low doses (6.25% protein replacement). Both genotoxicity assays (mammalian cell chromosome aberration assay and mammalian erythrocyte micronucleus test) returned negative results. These findings establish R. eutropha H16-derived SCP as a safe, nutritious, and sustainable protein source with considerable potential for feed and food applications, contributing to global food security and environmental sustainability. Full article

Background/Objectives: Chronic ethanol consumption damages erythrocyte membranes, reducing their deformability and disrupting erythropoiesis. Dopaminergic signalling plays an important role in regulating haematopoietic stem cells. However, the impact of genetic alterations in the dopamine system on the functional properties of erythrocytes and their resistance to the toxic effects of alcohol remains understudied. In this study, we evaluated haematological parameters and the sensitivity of erythrocytes to chronic ethanol exposure in DAT-HET rats (heterozygous for the dopamine transporter gene) compared to Wistar rats. Methods: Both lines of rats were subjected to chronic alcoholisation (10% ethanol for six months). Flow cytometry was used to assess reticulocyte production, intracellular esterase activity (cell viability), and membrane lipid asymmetry disruptions. Laser diffraction was used to evaluate osmotic resistance (deformability). Results: The partial knockout of the DAT gene significantly altered the baseline haematological profile, resulting in pronounced leukopenia, moderate thrombocytopenia, moderate erythrocyte macrocytosis, and reduced intracellular esterase activity. The consequences of chronic ethanol consumption in Wistar rats (anaemia, decreased reticulocyte count, increased osmotic rigidity of erythrocytes, and an increased proportion of annexin-positive cells) were similar to those observed in humans, thus confirming the validity of the experimental model. In contrast, chronic alcohol exposure did not significantly influence haematopoiesis parameters or erythrocyte biophysical properties in DAT-HET rats, indicating that even a 50% loss of the dopamine transporter strongly prevented ethanol’s damaging effects on blood cells. Conclusions: The marked contrast between Wistar rats (which develop alcohol-induced anaemia and erythrocyte dysfunction similar to humans) and DAT-HET rats (which remain largely unaffected) demonstrates that dopaminergic signalling plays a previously unrecognised role in determining erythrocyte sensitivity to alcohol toxicity. Full article

Background: Hemolysis and eryptosis of red blood cells (RBCs) contribute to chemotherapy-induced anemia, a marker of poor prognosis. Geraniol (GER) is an anticancer acyclic monoterpene alcohol found in several plant extracts, but a dearth of evidence exists regarding the potential toxicity of GER in RBCs. Methods: Hemolysis and eryptosis were evaluated using colorimetric and fluorescence-assisted cell-sorting methods, respectively. Phosphatidylserine (PS) exposure, loss of volume, and intracellular Ca²⁺ were measured by annexin-V-FITC, forward scatter (FSC), and Fluo4/AM staining. Cells were also examined by electron microscopy to identify membrane blebbing and by the Westergren method to assess erythrocyte sedimentation rate (ESR). Results: In a concentration-responsive fashion, GER induced hemolysis and PS exposure in addition to elevated ESR. GER-induced cell death was characterized by reduced FSC, membrane blebs, and increased Fluo4 fluorescence. Ca²⁺ deprivation prevented eryptosis, whereas concurrent Ca²⁺ deprivation and membrane depolarization prevented hemolysis, eryptosis, and cell shrinkage. Furthermore, whereas inhibition of cyclooxygenase (COX), casein kinase 1α (CK1α), or Rac1 GTPase ameliorated eryptosis and hemolysis, the latter was only prevented by caspase, nitric oxide synthase, or serine palmitoyltransferase inhibition. Exclusive reversal of eryptosis was rather only achieved in the presence of either caffeine or adenine. Conclusions: GER is a novel stimulator of hemolysis and eryptosis, an activity mediated through membrane hyperpolarization following Ca²⁺ elevation. In parallel, GER seems to involve the COX/CK1α/Rac1 GTPase axis to trigger its cytotoxic effects. Targeting the identified mechanisms in combination therapy may attenuate the off-target toxicity of GER and enhance its specificity to cancer cells. Full article

Red blood cells represent a widely used cellular model in cytotoxicity studies, particularly in hemocompatibility assessments. As enucleated cells, which are abundant and easily accessible in both humans and animals, red blood cells allow for rapid, reproducible, and low-cost evaluation of the toxicity of bioactive compounds, whether natural, synthetic, or nanoparticulate. From a functional perspective, the red blood cell membrane is highly sensitive to physical and chemical environmental changes (osmolarity, temperature, pH, and the presence of oxidizing agents). This sensitivity makes red blood cells an effective biosensor for detecting membrane damage, hemolysis, oxidative stress, methemoglobin formation, and aggregation processes. Therefore, in vitro tests using red blood cells allow for the preliminary evaluation in preclinical development, particularly for the early screening of cytotoxicity, membrane-disruptive effects, and hemocompatibility of small molecules, nanomaterials, and blood-contacting biomaterials. These techniques include hemocompatibility tests, evaluation of oxidative and osmotic damage, and evaluation of erythrocyte aggregation and function. However, the use of red blood cells as a cytotoxicity model also has significant limitations. As anucleate cells, erythrocytes lack organelles such as nuclei, mitochondria, or lysosomes, which prevents the evaluation of their effects on key intracellular processes such as protein synthesis, cell signaling, apoptosis, or endoplasmic reticulum stress. This lack of cellular complexity limits their usefulness as a sole model in studies of systemic toxicity or tissue-specific cytotoxicity. These tools offer an effective preliminary approach to anticipating risks in biomedical and pharmacological research. Full article

The essential oil or extract of Lemon myrtle (Backhousia citriodora F. Muell.), belonging to the family Myrtaceae and the genus Backhousia, exhibits anti-inflammatory and antioxidant properties. However, limited information exists on the safety of water extracts from its leaves. The present study aimed to assess the safety of lemon myrtle water extract as a functional food by performing genotoxicity studies and repeated-dose oral toxicity. Although the bacterial reverse mutation test (Ames test) yielded positive results, in vivo mammalian erythrocyte micronucleus and alkaline comet assays yielded negative results. In a 28-day oral toxicity study, the extract was orally administered to male and female Crl:CD rats at doses of 0, 250, 500, and 1000 mg/kg bw/day. Notably, the extract induced no adverse effects, and the no-observed-adverse-effect level was 1000 mg/kg bw/day in male and female rats. Despite its genotoxicity in vitro, the extract did not exhibit genotoxicity in vivo. Moreover, no signs of toxicity were observed in the general toxicity study. Overall, these results suggest that lemon myrtle water extract does not pose a substantive genotoxic risk at practical oral exposure levels. Full article

The fish liver, as the main detoxification organ, is highly susceptible to xenobiotic exposure, often resulting in various hepatopathies. The cytochrome P450 system plays a central role in xenobiotic metabolism, with cytochrome P450 reductase (CYPOR) supplying the electrons required for CYP enzyme activity. This study aimed to evaluate the relationship between the ecological state of a reservoir and fish health, including CYPOR levels, through hematological, bacteriological, and histological analyses. Samples of water and fish were collected from 12 littoral sites of Lake Ladoga. A total of 1360 specimens of fish from carp (Cyprinidae) and perch (Percidae) families were examined. For histological examination and CYPOR level determination, we selected 40 specimens using a blind randomization method. This sample size was sufficient for statistical analyses. Hematological smears were stained with azure eosin; bacteriological cultures were grown on multiple media; liver samples were stained with hematoxylin and eosin and Sudan III. CYPOR levels in liver homogenates were measured by ELISA-test. Physical and hydrochemical analyses indicated a high pollution level in the littoral zones. Isolated bacterial species were non-pathogenic but exhibited broad antibiotic resistance. Hematological evaluation revealed erythrocyte vacuolization and anisocytosis. Histological analysis showed marked fatty degeneration in hepatocytes, indicating toxic damage. CYPOR concentrations ranged from 0.3–0.4 ng/mL in healthy fish to 5–6 ng/mL in exposed specimens, showing strong correlation between environmental influence and enzyme activity. These findings demonstrate the potential of CYPOR as a sensitive biomarker for biomonitoring programs. The integrated methodological approach provides a model for assessing aquatic ecosystem health and identifying zones requiring priority remediation. Full article

Zinc oxide nanoparticles (ZnO NPs) are increasingly applied in medicine, cosmetics, and environmental technologies, yet their interactions with blood cells remain poorly understood, raising cross-species safety concerns. Using frog (nucleated) and human (anucleate) erythrocytes as comparative models, we show that cellular architecture fundamentally shapes responses to ZnO NPs exposure. Human erythrocytes exhibited a dose-dependent progression from membrane deformation to eryptosis and hemolysis, reflecting the pronounced vulnerability of anucleate cells. In contrast, frog erythrocytes sustained nuclear DNA damage while largely preserving membrane integrity, highlighting the protective or reparative role of the nucleus. Molecular docking revealed energetically favorable interactions of ZnO NPs with ERα-LBD and DNA (ΔG = −4.28 and −5.68 kcal/mol, respectively), while quantum chemical analyses indicated electron-accepting properties and a narrow HOMO–LUMO gap, suggesting efficient macromolecular interactions and intracellular ROS generation. Together, these findings demonstrate that the presence of a nucleus shifts the primary target of nanoparticle toxicity from membrane to genome, providing novel mechanistic insights. This comparative study offers a robust framework for understanding nanomaterial reactivity across taxa and informs One Health-oriented risk assessments. Full article