Toxins

12 pages, 3677 KB

Open AccessArticle

Identification of Unknown Biological Toxin Proteins Using Mass Spectrometry: A Case Study on De Novo Sequencing of Ricin

by Yubo Song, Hao Wang, Junjie Wen, Jiale Xu, Siyu Zhu, Fuli Wang and Yongqian Zhang

Toxins 2025, 17(11), 564; https://doi.org/10.3390/toxins17110564 - 18 Nov 2025

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Abstract

Background: The rapid and reliable identification of unknown or highly variable biological toxin proteins, such as the potent Ricin toxin, remains a critical challenge in biodefense and public security. Methods: To address this, we developed a Heuristic De Novo Sequencing (HDPS) strategy, which [...] Read more.

Background: The rapid and reliable identification of unknown or highly variable biological toxin proteins, such as the potent Ricin toxin, remains a critical challenge in biodefense and public security. Methods: To address this, we developed a Heuristic De Novo Sequencing (HDPS) strategy, which combines multiple enzymatic and microwave-assisted acid hydrolysis to generate diverse peptides, followed by a two-stage assembly process integrating de novo sequencing with homology-based database searching for robust error correction. Results: When applied to Ricin, this approach achieved 100% sequence coverage for both its A and B chains, with amino acid-level accuracies of 98.13% and 98.47%, respectively, and successfully corrected potential sequencing ambiguities. Conclusions: These results demonstrate that HDPS is a highly accurate and effective method for the de novo sequencing of full-length proteins, making it particularly valuable for characterizing unknown or mutated toxins in the absence of comprehensive reference databases. Full article

(This article belongs to the Special Issue Analytical Novelties and Challenges for the Detection of Natural Toxins)

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17 pages, 6436 KB

Open AccessArticle

Cytotoxic Effects Induced by Combined Exposure of the Patulin, Ochratoxin A, and Acetamiprid to HK-2 and SK-N-SH Cell Lines

by Zuoyin Zhu, Hanke Zhao, Xianli Yang, Dongxia Nie, Junhua Yang and Zheng Han

Toxins 2025, 17(11), 563; https://doi.org/10.3390/toxins17110563 - 18 Nov 2025

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Abstract

Patulin (PAT), ochratoxin A (OTA), and acetamiprid (ACM) are common food contaminants that frequently co-occur in agricultural products, raising concerns over their cumulative health risks. This study is the first to systematically assess the combined cytotoxic effects of PAT, OTA, and ACM using [...] Read more.

Patulin (PAT), ochratoxin A (OTA), and acetamiprid (ACM) are common food contaminants that frequently co-occur in agricultural products, raising concerns over their cumulative health risks. This study is the first to systematically assess the combined cytotoxic effects of PAT, OTA, and ACM using combination index (CI) and dose reduction index (DRI) models in HK-2 and SK-N-SH cells. All three compounds exhibited dose-dependent toxicity, with potency ranked as PAT > OTA > ACM. In HK-2 cells, PAT+OTA and OTA+ACM showed primarily antagonistic interactions, with synergism observed at low doses. PAT+ACM displayed exposure time-dependent additive effects, while the ternary mixture was mostly antagonistic, with OTA being the dominant contributor. In SK-N-SH cells, most combinations were antagonistic; however, OTA+ACM showed dose-dependent shifts, and the triple mixture transitioned from antagonism to synergism at higher concentrations. OTA and ACM were identified as the main toxicity drivers in all combinations. These findings highlight dose- and cell-specific interactions and underscore the importance of cumulative risk assessment of co-occurring mycotoxins and pesticides in food safety regulation. Full article

(This article belongs to the Special Issue Mycotoxin Co-Contamination：Cooccurrence, Interaction, Toxicological Mechanisms and, Risk Assessment in Agri-Products, Food and Feed)

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18 pages, 5231 KB

Open AccessArticle

Trace Aflatoxins Extraction in Pistachio, Maize and Rice Based on β-Cyclodextrin-Doped Cu-Carboxylated Graphene Oxide Nanocomposite

by Amr A. Yakout, Wael H. Alshitari, Hassan M. Albishri, Faten M. Ali Zainy and Adel M. Alshutairi

Toxins 2025, 17(11), 562; https://doi.org/10.3390/toxins17110562 - 17 Nov 2025

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Abstract

Aflatoxins remain among the most challenging food contaminants to monitor due to their structural diversity, low abundance, and the chemical complexity of cereal- and nut-based matrices. In this study, a multifunctional Cu/β-cyclodextrin@carboxylated graphene oxide (Cu/β-CD@CGO) nanocomposite was synthesized through a green, two-step procedure [...] Read more.

Aflatoxins remain among the most challenging food contaminants to monitor due to their structural diversity, low abundance, and the chemical complexity of cereal- and nut-based matrices. In this study, a multifunctional Cu/β-cyclodextrin@carboxylated graphene oxide (Cu/β-CD@CGO) nanocomposite was synthesized through a green, two-step procedure and employed as a high-affinity nanosorbent for trace extraction of AFB₁, AFB₂, AFG₁, and AFG₂. The architecture integrates three complementary components: β-cyclodextrin for inclusion-driven molecular recognition, copper nanoparticles that establish coordination interactions with lactone-bearing aflatoxins, and CGO nanosheets that supply extensive π-rich surfaces and abundant carboxyl functionalities. Comprehensive characterization (FTIR, Raman, XPS, SEM, EDX-mapping, and HRTEM) confirmed the formation of a uniform, porous hybrid network. Under optimized d-SPE conditions, the nanocomposite enabled quantitative recovery (92.0–108.5%) of aflatoxins from pistachio, maize, and rice extracts while achieving sub-ng kg⁻¹ detection limits and excellent reproducibility. The results demonstrate that the Cu/β-CD@CGO platform provides a robust, selective, and sustainable alternative to conventional immunoaffinity or polymeric sorbents, offering strong potential for routine surveillance of aflatoxins in complex food systems. Full article

(This article belongs to the Section Mycotoxins)

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18 pages, 2788 KB

Open AccessArticle

Venom Peptides Across Asian and American Tarantulas Utilize Dual Pharmacology to Target Activation and Fast Inactivation of Voltage-Gated Sodium Channels

by Amatulla S. Nashikwala, Charan Kotapati, David A. Eagles, Richard J. Lewis and Fernanda C. Cardoso

Toxins 2025, 17(11), 561; https://doi.org/10.3390/toxins17110561 - 14 Nov 2025

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Abstract

Spider-derived venoms are a rich source of cystine knot peptides with immense therapeutic potential. Many of these peptides exert unique biological activities through the modulation of ion channels, including of human voltage-gated sodium (Na_V1.1–Na_V1.9) channels. Na_V channel subtypes [...] Read more.

Spider-derived venoms are a rich source of cystine knot peptides with immense therapeutic potential. Many of these peptides exert unique biological activities through the modulation of ion channels, including of human voltage-gated sodium (Na_V1.1–Na_V1.9) channels. Na_V channel subtypes have diverse functions determined by their tissue and cellular distribution and biophysical properties, and are pathophysiology mediators in various diseases. Therefore, Na_Vs are central in studies of human biology. This work investigated the pharmacological properties of venom of the Thai theraphosid Ornithoctonus aureotibialis on Na_V channels. We discovered a predominant venom peptide named Oa1a and assessed its pharmacological properties across human Na_V channel subtypes. Synthetic forms of the peptide Oa1a showed preferential inhibition of Na_V1.1 and Na_V1.7, while recombinant Oa1a displayed a preference for inhibiting Na_V1.2, Na_V1.6, and Na_V1.7. Interestingly, all versions of Oa1a peptides exerted dual pharmacological effect by reducing the peak current and slowing fast inactivation of Na_V1.3, consistent with Oa1a having more than one binding site on Na_V channels. Such complex pharmacology was previously observed for a venom peptide in a Central American and Costa Rican tarantula, suggesting a conserved mechanism of action amongst these geographically distinct species. However, Oa1a lacked activity in the T-type channels observed in the tarantula peptide from Central America. Structure–function relationships investigated using molecular modelling showed that the dual pharmacology is driven by a conserved mechanism utilizing a mix of aromatic and charged residues, while the T-type activity appears to require additional charged residues in loop 2 and fewer positive charges in loop 4. Future structure–activity relationship studies of Oa1a will guide the development of pharmacological tools as well as next-generation drugs to treat Na_V channel dysfunction associated with neurological disorders. Full article

(This article belongs to the Section Animal Venoms)

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20 pages, 3294 KB

Open AccessArticle

Brevetoxin Dynamics and Bioavailability from Floc Following PAC-Modified Clay Treatment of Karenia brevis Blooms

by Nicholas R. Ohnikian, Christopher D. Sibley, R. Ben Freiberger, Kristen N. Buck, Alyssa Myers, Samantha Harlow, Donald M. Anderson, Richard Pierce and Jennifer H. Toyoda

Toxins 2025, 17(11), 560; https://doi.org/10.3390/toxins17110560 - 13 Nov 2025

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Abstract

Harmful algal blooms (HABs) caused by the dinoflagellate Karenia brevis present serious ecological and public health concerns due to the production of brevetoxins (BTX). Clay flocculation and sedimentation of cells, particularly with polyaluminum chloride (PAC)-modified clays, is a promising HAB mitigation approach. This [...] Read more.

Harmful algal blooms (HABs) caused by the dinoflagellate Karenia brevis present serious ecological and public health concerns due to the production of brevetoxins (BTX). Clay flocculation and sedimentation of cells, particularly with polyaluminum chloride (PAC)-modified clays, is a promising HAB mitigation approach. This study evaluated the efficacy of Modified Clay-II (MCII), a PAC-modified kaolinite clay, in reducing K. brevis cell abundance in mesocosm experiments and examined the bioavailability of BTX potentially released from settled floc back into the water column and sediment over the first 72 h after treatment. Additionally, we quantified trace metals in benthic clams (Mercenaria mercenaria) exposed to the floc post-treatment to assess metal accumulation and potential toxicological effects from MCII application. MCII treatment (0.2 g/L) resulted in a 91% reduction in K. brevis cell density and a 50% decrease in waterborne brevetoxins after 5 h. Brevetoxins accumulated in sediment post-flocculation, with BTX-B5 emerging as the dominant congener. Clams exposed to MCII-treated floc showed comparable tissue BTX levels to controls and significantly elevated aluminum concentrations, though without mortality. The aluminum accumulations in this study do not raise concerns for the health of the clams or the humans who eat them, given other dietary exposures. These findings support the potential of MCII for HAB mitigation while underscoring the need for further evaluation of exposure risks to all benthic species. Full article

(This article belongs to the Special Issue Managing and Preventing Harmful Algal Blooms (HABs): Do We Know Enough? (2nd Edition))

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23 pages, 11689 KB

Open AccessArticle

Visual Tracking and Organ Targeting of Naja atra and Deinagkistrodon acutus Venoms in Mice

by Shaocong Hu, Manqi Xiao, Ningjing Jiang, Ziyan Zhang, Qiuju Jia, Yi Zhou, Xin Liu and Ming Liao

Toxins 2025, 17(11), 559; https://doi.org/10.3390/toxins17110559 - 13 Nov 2025

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Abstract

In China, bites caused by the Naja atra and Deinagkistrodona acutus are the most common types of snakebites. While the functional characteristics of the two venom components have been well documented, their in vivo metabolic pathways, target organ distribution patterns, and dynamic pharmacokinetic [...] Read more.

In China, bites caused by the Naja atra and Deinagkistrodona acutus are the most common types of snakebites. While the functional characteristics of the two venom components have been well documented, their in vivo metabolic pathways, target organ distribution patterns, and dynamic pharmacokinetic profiles remain less explored. This study established a murine envenoming model through CY7-SE labeling of Naja atra and Deinagkistrodon acutus venoms. The real-time in vivo absorption and biodistribution of venoms were dynamically monitored via fluorescence imaging, with subsequent proteomic profiling to characterize organ-specific toxin targeting patterns. Gel filtration chromatography and HPLC analyses validated labeling efficiency at ratios of 0.1 mg CY7-SE per 1 mg Naja atra venom and 0.075 mg CY7-SE per 1 mg Deinagkistrodon acutus venom, with electrophoretic confirmation of protein integrity and preserved 740 nm fluorescence excitation. Acute toxicity assays demonstrated no significant difference in LD₅₀ lethality between labeled and native venoms (p > 0.05). The intoxication models revealed species-specific pathophenotypes, i.e., CY7-Naja atra venom induced systemic weakness, tachypnea, and inflammatory necrosis in lung, myocardium, and liver, whereas CY7-Deinagkistrodon acutus venom provoked hemorrhagic diathesis. Both models exhibited marked leukocytosis, transaminitis, and elevated creatinine levels (p < 0.05). Fluorescence tracing uncovered distinct biodistribution kinetics: Deinagkistrodon acutus venom achieved peak organ accumulation at 3 h with rapid dissemination (24 h injection-site retention: 12.61%), contrasting with Naja atra venom’s delayed 6 h peak and prolonged renal sequestration (24 h injection-site retention: 60.9%). Target organ proteomic profiling identified Deinagkistrodon acutus-enriched thrombin-like enzymes and metalloproteinases in lung/liver/spleen, while Naja atra venom predominantly accumulated renal acidic phospholipase A₂ and weakly neurotoxic NNAM2. Full article

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12 pages, 438 KB

Open AccessArticle

Early Botulinum Toxin Type A Injection May Improve Motor Recovery in Patients with Post-Stroke Spasticity: A Secondary Analysis from a Longitudinal Cohort Study

by Alessandro Picelli, Andrea Santamato, Michela Cosma, Alessio Baricich, Carmelo Chisari, Marzia Millevolte, Cristina Del Prete, Ilenia Mazzù, Rita Di Censo, Nicola Smania and Mirko Filippetti

Toxins 2025, 17(11), 558; https://doi.org/10.3390/toxins17110558 - 13 Nov 2025

Viewed by 774

Abstract

Spasticity after stroke impairs motor control, delays recovery, and reduces quality of life. Botulinum toxin type A is the first-line treatment, but it is often administered in the chronic phase, potentially limiting its impact on rehabilitation. Emerging evidence suggests that earlier treatment may [...] Read more.

Spasticity after stroke impairs motor control, delays recovery, and reduces quality of life. Botulinum toxin type A is the first-line treatment, but it is often administered in the chronic phase, potentially limiting its impact on rehabilitation. Emerging evidence suggests that earlier treatment may enhance recovery, though functional benefits remain uncertain. We conducted a secondary analysis of a multicenter, open-label, longitudinal cohort study to investigate whether the timing of the first botulinum toxin type A injection influences outcomes in post-stroke patients naïve to this treatment. All participants received botulinum toxin injections combined with conventional rehabilitation. Assessments were performed at baseline and at 4, 12, and 24 weeks post-injection. The primary outcome was muscle tone; secondary outcomes included motor strength, sensorimotor recovery, and global disability. Statistical analyses used mixed-effects models and trend tests. Patients treated within 90 days of stroke onset showed greater reductions in spasticity at 4 and 12 weeks compared with later treatment. Despite having more severe baseline impairments, early treated patients demonstrated faster and more pronounced improvements in upper-limb strength, sensorimotor recovery, and global disability. Early toxin administration is associated with enhanced reduction in spasticity and improved motor recovery, particularly in patients with severe initial deficits. Full article

(This article belongs to the Special Issue Botulinum Toxin: Advancing Treatments for Spasticity)

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19 pages, 2162 KB

Open AccessArticle

Optimization by the 4S Sequential Experimental Design Process of a Competitive Lateral Flow Immunoassay Device for the Detection of Aflatoxin B1

by Simone Cavalera, Sofia Stanzani, Thea Serra, Valentina Testa, Fabio Di Nardo, Claudio Baggiani and Laura Anfossi

Toxins 2025, 17(11), 557; https://doi.org/10.3390/toxins17110557 - 13 Nov 2025

Viewed by 454

Abstract

Aflatoxin B1 (AFB1) is a highly toxic and carcinogenic compound produced by certain fungi (e.g., Aspergillus flavus and Aspergillus parasiticus). Rapid and ultra-sensitive detection methods for AFB1 in various commodities are in high demand. This study aimed to enhance the sensitivity of [...] Read more.

Aflatoxin B1 (AFB1) is a highly toxic and carcinogenic compound produced by certain fungi (e.g., Aspergillus flavus and Aspergillus parasiticus). Rapid and ultra-sensitive detection methods for AFB1 in various commodities are in high demand. This study aimed to enhance the sensitivity of a competitive lateral flow immunoassay (LFIA) for AFB1 detection by leveraging a previously developed experimental design strategy, named 4S. This approach comprises four phases—START, SHIFT, SHARPEN, and STOP—and involves the analysis of two reference conditions: NEG (0 ng/mL AFB1) and POS (1 ng/mL AFB1). By generating and overlaying response surfaces, regions of optimal NEG signal and POS/NEG signal ratio (IC%) were identified. Four variables were optimized: two related to the labeled antibody (its concentration and antibody-to-label ratio) and two to the competitor antigen (its concentration and hapten-to-protein ratio). An initial design defined the parameter space, while three subsequent designs did not yield further improvements in sensitivity. A strong anti-correlation was observed between the IC% and competitor parameters. The optimized LFIA-1 exhibited enhanced sensitivity, achieving a limit of detection of 0.027 ng/mL compared to 0.1 ng/mL for the original device. Additionally, the amount of expensive antibody required for device fabrication was reduced by around a factor of four. Full article

(This article belongs to the Section Mycotoxins)

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22 pages, 2245 KB

Open AccessReview

Adsorption–Degradation Integrated Approaches to Mycotoxin Removal from Food Matrices: A Comprehensive Review

by Xiyu Yang, Mingjian Yao, Wenchao Liao and Xiaoyang Li

Toxins 2025, 17(11), 556; https://doi.org/10.3390/toxins17110556 - 12 Nov 2025

Viewed by 803

Abstract

Mycotoxin contamination is a crucial issue in food safety. However, the removal of trace amounts of mycotoxins from complex food and feed matrices without significant loss of nutritional and flavor quality remains a significant challenge. The integrated adsorption–catalysis strategy involves immobilizing catalytic modules [...] Read more.

Mycotoxin contamination is a crucial issue in food safety. However, the removal of trace amounts of mycotoxins from complex food and feed matrices without significant loss of nutritional and flavor quality remains a significant challenge. The integrated adsorption–catalysis strategy involves immobilizing catalytic modules onto adsorption materials, enabling in situ degradation while enriching the mycotoxins. This approach can significantly reduce the dosage of detoxification agents and achieve efficient removal of trace mycotoxins in food. This review provides an overview of adsorbents with enrichment capabilities and their applications in the targeted removal of mycotoxins from food. The adsorption–degradation coupled systems are categorized into the following two main types: adsorption–photocatalysis coupled systems and adsorption–biocatalysis coupled systems. The review introduces recent advances in the design of bifunctional catalysts, focusing on their synergistic mechanisms and practical applications for detoxifying various mycotoxins in food matrices. Finally, the review discusses current industrial challenges and offers insights into future directions for this field. Full article

(This article belongs to the Special Issue Advances in Detection and Removal of Mycotoxins in Food)

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20 pages, 3842 KB

Open AccessReview

Deoxynivalenol and Its Metabolites: Contamination, Metabolism, and Toxicity

by Yukai Lin, Ruibiao Wang, Suxian Liu, Hanqing Zhao, Bo Wen, Songbiao Chen, Rongxian Guo, Lei Wang, Xiaojing Xia, Yanzhao Xu and Ke Ding

Toxins 2025, 17(11), 555; https://doi.org/10.3390/toxins17110555 - 11 Nov 2025

Viewed by 713

Abstract

Deoxynivalenol (DON), a toxic secondary metabolite produced by Fusarium graminearum in infected cereal crops, has emerged as a major global contaminant in food and feed due to its stable physicochemical properties and resistance to degradation during conventional processing. This contamination poses a serious [...] Read more.

Deoxynivalenol (DON), a toxic secondary metabolite produced by Fusarium graminearum in infected cereal crops, has emerged as a major global contaminant in food and feed due to its stable physicochemical properties and resistance to degradation during conventional processing. This contamination poses a serious threat to livestock production and animal health. This review provides a comprehensive overview of the current status of DON contamination, its transmission through the food chain, metabolic pathways in animals, and the comparative toxicity of its metabolites. Furthermore, we analyze DON-induced toxic effects, including acute toxicity, cytotoxicity, immunotoxicity, neurotoxicity, gastrointestinal toxicity, and hepatotoxicity. By integrating domestic and international regulatory thresholds with current mitigation strategies, we highlight future research directions focusing on biodegradation technologies and genetic regulation approaches to alleviate DON contamination in livestock feeds. Advancing efficient DON-degradation strategies could open new avenues for sustainable feed management and mycotoxin detoxification technologies. Full article

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15 pages, 722 KB

Open AccessArticle

Exposure Type and Duration Determine Ecotoxicological Effects of Cyanobacteria Anatoxins on the Benthic Amphipod Hyalella azteca

by Isabelle Kamalani Yogeshwar, Erwin J. J. Kalis, Juergen Geist and Sebastian Beggel

Toxins 2025, 17(11), 554; https://doi.org/10.3390/toxins17110554 - 7 Nov 2025

Viewed by 640

Abstract

Cyanobacteria can pose a threat to aquatic organisms by their ability to produce toxins such as neurotoxic anatoxins. Although cyanobacteria and their effects on aquatic fauna have been a research focus for a long time, the interactions between benthic cyanobacteria and benthic invertebrates [...] Read more.

Cyanobacteria can pose a threat to aquatic organisms by their ability to produce toxins such as neurotoxic anatoxins. Although cyanobacteria and their effects on aquatic fauna have been a research focus for a long time, the interactions between benthic cyanobacteria and benthic invertebrates are still largely unknown, especially with regard to how invertebrates cope with cyanotoxins which they are exposed to in their habitat. This study characterizes the effects of anatoxins on the benthic macroinvertebrate Hyalella azteca. In a first test, organisms were exposed to synthetically produced anatoxins dissolved in the ambient aqueous phase. In a second test, organisms were exposed to natural anatoxins within intact Tychonema cells as their sole food source. Over 10 days of aqueous exposure to anatoxins, survival of H. azteca was not affected, even at the highest nominal concentrations of 587.37 µg/L ATX and 590.31 µg/L dhATX. Over 42 days of dietary exposure to natural anatoxins, H. azteca readily accepted Tychonema as a food source. Survival, growth, reproductive success and storage compound concentrations (glucose, glycogen, lipid and protein) in the organisms’ tissue, all assessed in the same individuals, were reduced. These findings suggest that the ecotoxicological effects of anatoxins on aquatic invertebrates not only depend on their concentration, but even more so on the type and duration of exposure. Furthermore, cyanobacteria like Tychonema seem to be insufficient as source of energy if they represent the only available food source. Full article

(This article belongs to the Section Marine and Freshwater Toxins)

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13 pages, 3153 KB

Open AccessArticle

Evaluation of Activity of Pro- and Anti-Inflammatory Mediators and Nitrosative Stress in Liver Tissue of Wild Boars (Sus scrofa) Positive for Zearalenone (ZEN) Contamination in Campania Region, Southern Italy

by Sara Damiano, Consiglia Longobardi, Evaristo Di Napoli, Valentina Iovane, Francesco Ferrucci, Giuseppe Rizzo, Antonio Raffaele, Antonio Rubino, Valeria Russo and Roberto Ciarcia

Toxins 2025, 17(11), 553; https://doi.org/10.3390/toxins17110553 - 5 Nov 2025

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Abstract

Zearalenone (ZEN) is a mycotoxin commonly produced by Fusarium species and is often found in food and feed. It has been linked to reproductive problems in livestock and, less frequently, to hyperestrogenic effects in humans. However, information regarding the impact of ZEN on [...] Read more.

Zearalenone (ZEN) is a mycotoxin commonly produced by Fusarium species and is often found in food and feed. It has been linked to reproductive problems in livestock and, less frequently, to hyperestrogenic effects in humans. However, information regarding the impact of ZEN on wild boars (Sus scrofa) remains scarce, despite this species being among the most frequently hunted game animals in Italy. The aim of this study was to assess the impact of ZEN on the hepatic system by examining nitrosative stress markers and the balance between pro- and anti-inflammatory cytokines in wild boars hunted in various areas of the Avellino province (Campania region, Italy) during the 2021–2022 hunting season. The findings revealed that exposure to ZEN was linked to a marked rise in both pro- and anti-inflammatory mediators, except for IL-10, which did not increase significantly. In addition, ZEN stimulated the expression of inducible nitric oxide synthase (iNOS), which, in turn, led to elevated nitric oxide (NO) concentrations in the liver. The immunohistochemical analysis revealed a predominance of CD3-positive T-cells in the hepatic inflammatory infiltrate of ZEN-exposed wild boars, highlighting the importance of structured wildlife monitoring to protect food safety and safeguard human and animal health. Full article

(This article belongs to the Special Issue Mycotoxins—Biomonitoring and Exposure)

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16 pages, 1305 KB

Open AccessArticle

Occurrence and Levels of Emerging Alternaria Mycotoxins Detected in Spices and Herbs Marketed in Italy

by Katia Gialluisi, Maria Giovanna Nicoletti, Nada El Darra, Michele Solfrizzo and Lucia Gambacorta

Toxins 2025, 17(11), 552; https://doi.org/10.3390/toxins17110552 - 5 Nov 2025

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Abstract

Emerging Alternaria mycotoxins tenuazonic acid (TeA), alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT) and tentoxin (TEN) were detected in samples of spices and herbs. A total of 92 samples purchased in Italian markets were analyzed by using a UPLC-MS/MS method. TeA was [...] Read more.

Emerging Alternaria mycotoxins tenuazonic acid (TeA), alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT) and tentoxin (TEN) were detected in samples of spices and herbs. A total of 92 samples purchased in Italian markets were analyzed by using a UPLC-MS/MS method. TeA was the predominant mycotoxin with the highest percentage of positive samples (53%), followed by AME, AOH and TEN with overall means of 801.5, 2.4, 6.2 and 10.8 µg/kg, respectively. All samples were negative to ALT. The spices with higher levels of mycotoxins were flax seeds, paprika, red chili and licorice; regarding herbs, they were basil, sage and oregano. While TeA was found to be the most abundant mycotoxin equally in Italian and Lebanese samples, AOH and AME showed significantly lower levels in Italian samples, as Lebanese samples contained concentrations up to 14 times higher. Considering the mean levels of TeA in samples, the daily consumption of >8.7 g of flax seeds, >9.3 g of paprika and >5.8 g of red chili will exceed the threshold of toxicological concern (TTC) of TeA, which is 1500 ng/kg bw for a day. The high percentage of positive samples and the high levels of some mycotoxins observed demonstrate the susceptibility of spices and herbs to Alternaria mycotoxin contamination. These results provide an overview of emerging Alternaria mycotoxins in spices and herbs marketed in Italy and represent a valuable starting point to assess human exposure and support future studies aimed at establishing safe limits to protect human health. Full article

(This article belongs to the Special Issue Risk Assessment of Mycotoxins: Challenges and Emerging Threats)

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19 pages, 2140 KB

Open AccessArticle

Efficacy of Dodonaea viscosa Extract and Its Chitosan-Based Nanoparticle Formulation on the Growth of Fusarium Strains and the Production of Deoxynivalenol and Moniliformin in Stored Wheat

by Hussein Ali Salman Alsahho, Sumer Horuz and Kevser Karaman

Toxins 2025, 17(11), 551; https://doi.org/10.3390/toxins17110551 - 5 Nov 2025

Viewed by 455

Abstract

Fusarium is considered one of the most important fungi that attack plants and cause serious diseases resulting in huge losses to crops, especially wheat. Fungicides have been used to control it, but they have drawbacks, including residues and toxicity to mammals, which encouraged [...] Read more.

Fusarium is considered one of the most important fungi that attack plants and cause serious diseases resulting in huge losses to crops, especially wheat. Fungicides have been used to control it, but they have drawbacks, including residues and toxicity to mammals, which encouraged researchers to find alternatives to these methods and materials. This study was conducted to find natural alternatives to the chemicals used as fungicides. The Dodonaea viscosa plant extract was evaluated as an extract (DVE) and nanoparticles (chitosan NPs loaded with DVE) to inhibit the growth of Fusarium spp. strains and production ability of Deoxynivalenol (DON) and Moniliformin (MON) mycotoxins. The wheat samples were taken from storage in eighteen different governorates in Iraq. Fusarium spp. strains were detected phenotypically, and seven strains were identified by using the polymerase chain reaction technique (PCR) as F. oxysporum, F. pseudograminearum and F. chlamydosporum. DVE effectively inhibited the growth of Fusarium spp. strains at three different concentrations (0.5, 1.0, and 1.5%) on PDA. The highest percentage was 68.94% for F. oxysporum strain 5, and the lowest percentage was 22.58% for F. pseudograminearum strain 6 at a concentration of 1.5%. However, applying chitosan NPs loaded with DVE at a concentration of 0.75% effectively increased the inhibition rate. The treatment of chitosan NPs loaded with DVE played a role in inhibiting the percentage of mycotoxins produced. The highest percentage of inhibition of the DON toxin was recorded as 73.75% in Fusarium pseudograminearum strain 2, and the highest percentage of inhibition of the production of the (MON) toxin was 73.62% in isolate Fusarium chlamydosporum strain 8. Overall, this study highlights for the first time the potential of Dodonaea viscosa nano-formulation to suppress both fungal growth and mycotoxin biosynthesis, providing a sustainable and safe strategy for protecting stored grains. Full article

(This article belongs to the Special Issue Advances in Detection and Removal of Mycotoxins in Food)

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16 pages, 4013 KB

Open AccessArticle

OpiCa1 Modulates Cardiomyocyte Viability Through PI3K/Akt Inhibition with Minimal Systemic Impact Beyond RyR Targeting

by Xiaofen Ma, Xiaoyu Hua, Xiao Peng, Zhixiao Yang, Yi Wang, Qian Zhang, Lianbo Wang, Shumin Wang, Héctor H. Valdivia, Liang Xiao and Mei Wang

Toxins 2025, 17(11), 550; https://doi.org/10.3390/toxins17110550 - 4 Nov 2025

Viewed by 550

Abstract

Calcins represent a class of novel peptide ligands for ryanodine receptors (RyRs), demonstrating therapeutic potential against Ca²⁺ dysregulation-related cardiac diseases. Nevertheless, their biological effects beyond RyR modulation and underlying mechanisms remain unexplored. This study employed Opicalcin1 (OpiCa1), the most bioactive calcin member, [...] Read more.

Calcins represent a class of novel peptide ligands for ryanodine receptors (RyRs), demonstrating therapeutic potential against Ca²⁺ dysregulation-related cardiac diseases. Nevertheless, their biological effects beyond RyR modulation and underlying mechanisms remain unexplored. This study employed Opicalcin1 (OpiCa1), the most bioactive calcin member, revealing that while it reduced cytosolic Ca²⁺ in H9c2 cardiomyocytes, it concurrently diminished cell viability and promoted apoptosis. Transcriptomics and Western blot analyses identified suppression of the negatively regulatory PI3K/Akt pathway as the mechanistic basis. In acute/chronic in vivo studies, high-dose OpiCa1 (≥50 mg/kg i.v.) exhibited minimal impact on body weight, histopathology, and organ indices, while accompanied with subtle alterations in serum indicators, including slight elevations in AST, ALT, and LDH, alongside mild reductions in CK-MB and TBIL-Z. These findings unveil OpiCa1’s modulation on cardiomyocyte viability through PI3K/Akt inhibition with minimal systemic impact, providing new insights into non-RyR-mediated actions of calcins and critical toxicological support for developing calcin-based therapies targeting Ca²⁺-dysregulated cardiac pathologies. Full article

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12 pages, 5855 KB

Open AccessArticle

In Silico Binding Mode Analysis of Blarina Paralytic Peptides with the Human T-Type Ca Channel hCa_v3.2

by Nozomi Hara, Chihiro Sadamoto, Ryo Fukuoka, Yusuke Yano, Andres D. Maturana and Masaki Kita

Toxins 2025, 17(11), 549; https://doi.org/10.3390/toxins17110549 - 4 Nov 2025

Viewed by 468

Abstract

Blarina paralytic peptides (BPPs), neurotoxins from shrew saliva that paralyze mealworms, share high sequence similarity with human synenkephalin [1–53] (hSYN), a peptide released from proenkephalin together with opioid peptides that mediate analgesic and antidepressant effects in the brain. Both synthetic BPP2 and hSYN [...] Read more.

Blarina paralytic peptides (BPPs), neurotoxins from shrew saliva that paralyze mealworms, share high sequence similarity with human synenkephalin [1–53] (hSYN), a peptide released from proenkephalin together with opioid peptides that mediate analgesic and antidepressant effects in the brain. Both synthetic BPP2 and hSYN induce a hyperpolarizing shift in the human T-type voltage-gated calcium channel (hCa_v3.2) at sub-micromolar concentrations, although only BPP2 causes paralysis in insects. To gain insight into the functions of these insectivorous animal-specific neurotoxins and the largely uncharacterized brain peptides, we investigated the structure prediction of BPPs and SYNs and their interactions with hCa_v3.2. AlphaFold 3 modeling complemented available cryo-EM data and accurately reproduced the overall channel architecture; however, this inactivated-state model proved unsuitable for predicting agonistic binding of BPPs and SYNs. In contrast, docking simulations using an activated-state hCa_v3.2 homology model revealed distinct ligand-dependent differences in binding energies, affinity, and conformational flexibility. Notably, the C-terminal tail of BPPs—particularly its variable length and flexibility—was identified as a key determinant for the interactions with the S4 voltage-sensing domain of the channel. These findings provide new insights into the evolutionary adaptation of venom peptides in mammals and into potential therapeutic strategies targeting neurological disorders. Full article

(This article belongs to the Special Issue Venomics Insights into the Evolutionary Biology of Peptide Toxins in Marine and Terrestrial Organisms)

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14 pages, 943 KB

Open AccessArticle

Well-Being, Protein-Bound Toxins, and Dietary Fibre in Patients with Kidney Disease: Have We Been Missing the Obvious?

by Aruni Malaweera, Louis L. Huang and Lawrence P. McMahon

Toxins 2025, 17(11), 548; https://doi.org/10.3390/toxins17110548 - 4 Nov 2025

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Abstract

Aim: To explore the associations between protein-bound uraemic toxins (PBTs), fibre intake and patient-focused outcomes in patients on kidney replacement therapy. Background: Despite removal of small water-soluble uraemic toxins, dialysis patients continue to experience high morbidity and mortality. Recent evidence suggests strong associations [...] Read more.

Aim: To explore the associations between protein-bound uraemic toxins (PBTs), fibre intake and patient-focused outcomes in patients on kidney replacement therapy. Background: Despite removal of small water-soluble uraemic toxins, dialysis patients continue to experience high morbidity and mortality. Recent evidence suggests strong associations between PBTs and poorer patient outcomes and symptom burden. Reducing the generation of PBTs by increasing dietary fibre may be an alternate approach to better patient outcomes. Method: This was a cross-sectional study of haemodialysis (HD), peritoneal dialysis (PD) and kidney transplant patients to determine the associations between uraemic toxins [p-cresyl sulfate (PCS) and indoxyl sulfate (IS)], fibre intake and patient-focused outcomes, incorporating the Integrated Palliative Outcome Scale-Renal (IPOS-renal) and EQ-5D-5L to determine symptom burden and quality of life, while physical capacity was determined using the timed up and go(TUG) test and handgrip strength (HGS). Results: Ninety participants completed the study (n = 30 in each group). There was a correlation between PBTs and the IPOS-renal score, where higher toxin levels were associated with a higher symptom burden. This was the strongest for PCS, where the significance remained after accounting for age and co-morbidities (p < 0.05). Higher PBT levels were also associated with lower HGS (p < 0.05). There was a negative correlation between fibre intake and PBTs, serum PCS (r = −0.36, p < 0.05) and serum IS (r = −0.27, p < 0.05). Lower fibre intake was also associated with a higher symptom burden measured by the IPOS-renal (p < 0.05). Transplant patients consistently performed better, with a reduced symptom burden and improved physical ability compared to dialysis patients. Conclusion: PBTs were associated with symptom burden, and lower physical ability was associated with both PBTs and patient-focused outcomes, and this needs to be further investigated in larger studies. Full article

(This article belongs to the Special Issue The Role of Uremic Toxins in Comorbidities of Chronic Kidney Disease)

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13 pages, 377 KB

Open AccessArticle

OnabotulinumtoxinA to Prevent Chronic Migraine with Comorbid Bruxism: Real-World Data from the GRASP Study Group

by Andreas A. Argyriou, Emmanouil V. Dermitzakis, Maria Chondrogianni, Aikaterini Foska, Dimitrios Rikos, Georgia Xiromerisiou, Panagiotis Soldatos, Pantelis Litsardopoulos and Michail Vikelis

Toxins 2025, 17(11), 547; https://doi.org/10.3390/toxins17110547 - 3 Nov 2025

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Abstract

Background: This study, designed by the Greek Research Alliance for the Study of Headache and Pain (GRASP), sought to prospectively examine whether the treatment with two consecutive OnabotulinumtoxinA (BoNTA) cycles might improve the frequency and severity of chronic migraine (CM) with comorbid bruxism. [...] Read more.

Background: This study, designed by the Greek Research Alliance for the Study of Headache and Pain (GRASP), sought to prospectively examine whether the treatment with two consecutive OnabotulinumtoxinA (BoNTA) cycles might improve the frequency and severity of chronic migraine (CM) with comorbid bruxism. We also explored whether the potential BoNTA-related alleviation of bruxism can directly influence the improvements in migraine efficacy outcomes. Methods: A total of 58 CM patients with comorbid bruxism at baseline, attaining two consecutive (quarterly given) BoNTA cycles, were studied. The changes in bruxism-related pain were assessed with the 0–10 numeric scale PI-NRS. Bruxism was clinically diagnosed using the self-report Bruxscreen-Q questionnaire. Any phenotypic changes in bruxism, according to Bruxscreen-Q, from baseline (T0) to the last efficacy evaluation follow-up (T1), were analyzed and then compared. Migraine-related efficacy and disability outcomes, mostly mean headache days (MHD), were also compared between T0 and T1. Results: BoNTA exerted significant improvements in bruxism-related pain, with PI-NRS median scores being significantly reduced from 7 at T0 to 3 at T1 (p < 0.001). The rates of masseter hypertrophy at T1 significantly dropped, compared to T0 (chi-square: 16; p < 0.001). Patients also self-reported significant improvements in the Bruxscreen-Q items at T1, compared to T0. At T1, 41/58 (70.7%) patients responded to BoNTA. The significant decrease in MHD frequency at T1 was positively correlated with improvements in bruxism-related pain severity (Pearson’s correlation: 0.710; p < 0.001). Conclusions: BoNTA exerts dual beneficial effects towards both the reduction of migraine frequency and the alleviation of bruxism-related pain and disability. Both of these effects seem closely interrelated in our study. Full article

(This article belongs to the Special Issue Botulinum Toxin Therapy for Diseases in the Oral and Maxillofacial Regions)

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13 pages, 1595 KB

Open AccessArticle

Characterization of Hottentotta judaicus Scorpion Venom: Toxic Effects and Neurobehavioral Modulation in Insect Models

by Rim Wehbe, Aline Karaki, Zeina Dassouki, Mohamad Rima, Adolfo Borges, Rabih Roufayel, Christian Legros, Ziad Fajloun and Zakaria Kambris

Toxins 2025, 17(11), 546; https://doi.org/10.3390/toxins17110546 - 3 Nov 2025

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Abstract

Scorpion venom is a rich source of diverse bioactive molecules with medicinal importance. While the venoms of many Buthidae scorpions have been extensively studied for their toxicity and therapeutic potential, Hottentotta judaicus scorpion venom (HjSV) remains poorly explored. In this study, using LC-ESI-MS, [...] Read more.

Scorpion venom is a rich source of diverse bioactive molecules with medicinal importance. While the venoms of many Buthidae scorpions have been extensively studied for their toxicity and therapeutic potential, Hottentotta judaicus scorpion venom (HjSV) remains poorly explored. In this study, using LC-ESI-MS, we show that HjSV has a complex composition. We find that HjSV has no significant cytotoxic effects on three human cancer cell lines, even at concentrations of up to 1000 µg/mL. However, it exerts a dose-dependent insecticidal effect against Drosophila melanogaster, a well-established genetic model organism, and two medically relevant mosquito species, Aedes albopictus and Culex pipiens. These findings highlight the venom’s selective activity and reveal a species-dependent susceptibility in insects, with mosquitoes being more sensitive than Drosophila. Furthermore, we show that at sub-lethal doses, HjSV alters D. melanogaster behavioral patterns, significantly reducing locomotor activity and increasing sleep duration. Altogether, our results provide new insights into the dual role of HjSV as both an insecticidal agent and behavioral modulator, shedding light on its ecological function in prey subduing and its potential application in pest control strategies. Full article

(This article belongs to the Special Issue Animals Venom in Drug Discovery: A Valuable Therapeutic Tool)

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15 pages, 1521 KB

Open AccessArticle

The Activities of Recombinant Botulinum Toxin A on Spared Nerve Injury-Induced Neuropathic Pain in a Diabetic Mice Model

by Akinyemi Ademola Omoniyi, Rasmus Eich Hammer, Sabrina Josefsen, Mette Richner, Stephane Lezmi, Christian Bjerggaard Vægter, Mikhail Kalinichev, Páll Karlsson and Jens Randel Nyengaard

Toxins 2025, 17(11), 545; https://doi.org/10.3390/toxins17110545 - 3 Nov 2025

Viewed by 661

Abstract

Diabetic neuropathy is characterized by nerve damage and chronic neuropathic pain and lacks effective treatment. Botulinum neurotoxin type A (BoNT/A), a neurotoxin with established therapeutic use in neurological disorders, has emerged as a potential analgesic agent. This study investigated the effects of a [...] Read more.

Diabetic neuropathy is characterized by nerve damage and chronic neuropathic pain and lacks effective treatment. Botulinum neurotoxin type A (BoNT/A), a neurotoxin with established therapeutic use in neurological disorders, has emerged as a potential analgesic agent. This study investigated the effects of a recombinant form of BoNT/A1 (rBoNT/A1) on neuropathic pain induced by spared nerve injury (SNI) in a diabetic mouse model. Thirty-two adult male C57BL/6JRj diabetic mice were subjected to SNI or sham surgery. Fourteen days post surgery, mice received an intraplantar dose of rBoNT/A1 or vehicle. Mechanical allodynia was assessed using von Frey filaments, and spinal cord and sciatic nerve tissues were analyzed via immunohistochemistry and transmission electron microscopy to evaluate glial activation, neurotransmitter receptor expression, and axonal morphology. The results demonstrated that rBoNT/A1 significantly alleviated mechanical allodynia and caused a marked reduction in Iba1-positive microglial activation in the spinal cord, whereas no significant changes were observed in astrocyte (GFAP) density or GABAAR subunit expression. Additionally, rBoNT/A1 treatment did not significantly alter axon diameter, myelin thickness, or C-fiber morphology. In conclusion, intraplantar administration of rBoNT/A1 reduced SNI-induced mechanical allodynia in diabetic mice, potentially by attenuating spinal microglial activation, supporting the therapeutic promise of rBoNT/A1 in managing diabetic neuropathic pain. Full article

(This article belongs to the Special Issue Botulinum Toxins: Beyond the SNAP-25 Cleavage and New Insights on Pain Relief Mechanisms)

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13 pages, 419 KB

Open AccessArticle

Annual and Seasonal Variations in Aflatoxin M1 in Milk: Updated Health Risk Assessment in Serbia

by Saša Krstović, Sandra Jakšić, Jelena Miljanić, Borislav Iličić, Milica Živkov Baloš, Darko Guljaš, Marko Damjanović and Igor Jajić

Toxins 2025, 17(11), 544; https://doi.org/10.3390/toxins17110544 - 2 Nov 2025

Viewed by 832

Abstract

Aflatoxin M1 (AFM1), a hepatocarcinogenic metabolite of aflatoxin B1, poses significant risks to human health through its presence in milk and dairy products. This study investigates AFM1 contamination in raw milk produced in Serbia from 2021 to 2025, assessing annual and seasonal variations [...] Read more.

Aflatoxin M1 (AFM1), a hepatocarcinogenic metabolite of aflatoxin B1, poses significant risks to human health through its presence in milk and dairy products. This study investigates AFM1 contamination in raw milk produced in Serbia from 2021 to 2025, assessing annual and seasonal variations and associated health risks. A total of 907 milk samples were analyzed using enzyme-linked immunosorbent assay (ELISA), revealing contamination in 70.1% of samples, with mean concentrations exceeding the EU regulatory limit of 50 ng/kg. Seasonal analysis identified the highest contamination levels during winter, attributed to increased use of contaminated feed during colder months. Health risk assessments estimated the daily intake of AFM1 and associated health risks, with high-exposure individuals showing notably reduced margins of safety. The research demonstrates the essential requirement for better feed quality management alongside enhanced regulatory oversight along with health programs that reduce AFM1 exposure in Serbian populations. Full article

(This article belongs to the Special Issue From Inhibiting Fungal Growth to Reducing Mycotoxin Content in Food and Feed: Challenges, Achievements, and Advanced Approaches)

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14 pages, 809 KB

Open AccessReview

Botulinum Toxin Type A for Pediatric Torticollis: A Review of Clinical Research

by Na-Yoen Kwon, Soo-Hyun Sung and Hyun-Kyung Sung

Toxins 2025, 17(11), 543; https://doi.org/10.3390/toxins17110543 - 1 Nov 2025

Viewed by 682

Abstract

Pediatric torticollis, predominantly resulting from congenital muscular torticollis, is characterized by unilateral shortening of the sternocleidomastoid muscle, leading to head tilt and limited cervical mobility. Conventional management primarily involves physical therapy and repositioning strategies, with most infants achieving full recovery. However, a subset [...] Read more.

Pediatric torticollis, predominantly resulting from congenital muscular torticollis, is characterized by unilateral shortening of the sternocleidomastoid muscle, leading to head tilt and limited cervical mobility. Conventional management primarily involves physical therapy and repositioning strategies, with most infants achieving full recovery. However, a subset of patients exhibits persistent symptoms despite conservative treatment. Botulinum toxin type A (BoNT-A) has emerged as a minimally invasive adjunct intervention that targets muscular hypertonicity by inhibiting acetylcholine release at neuromuscular junctions. This scoping review synthesizes clinical evidence from six studies, including randomized controlled trials and case reports, assessing the efficacy and safety of BoNT-A in pediatric torticollis. Results indicate consistent improvements in range of motion, head posture correction, and patient satisfaction, with rare and mild adverse events such as local bruising and transient muscle weakness. Despite promising outcomes, variability in dosing, injection protocols, and follow-up durations underscores the need for standardized treatment guidelines and further high-quality research. These findings support BoNT-A as a valuable therapeutic option for refractory pediatric torticollis, warranting integration into multidisciplinary care frameworks. Full article

(This article belongs to the Special Issue Natural Toxin–Based Therapeutics from Animals, Plants, and Marine Organisms: Clinical Evidence and Drug Development Perspectives)

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14 pages, 855 KB

Open AccessArticle

Beyond Glycemic Control: Concurrent GLP-1 Receptor Agonist Use Is Associated with Reduced Urinary Adverse Events Following OnabotulinumtoxinA Treatment in Non-Diabetic Adults with Overactive Bladder

by Muhammed A. M. Hammad, Sophia G. Quesada, Aimee L. Belczyk and Gamal M. Ghoniem

Toxins 2025, 17(11), 542; https://doi.org/10.3390/toxins17110542 - 1 Nov 2025

Viewed by 1489

Abstract

Semaglutide, a GLP-1 (glucagon-like peptide-1) receptor agonist, is widely prescribed for weight loss in non-diabetic populations. Given the link between obesity and overactive bladder (OAB), we explored whether GLP-1 use would improve adverse urinary events beyond its weight loss benefit for non-diabetic adults [...] Read more.

Semaglutide, a GLP-1 (glucagon-like peptide-1) receptor agonist, is widely prescribed for weight loss in non-diabetic populations. Given the link between obesity and overactive bladder (OAB), we explored whether GLP-1 use would improve adverse urinary events beyond its weight loss benefit for non-diabetic adults undergoing onabotulinumtoxin A (BTX-A) treatment for OAB. Using the TriNetX database, we conducted a retrospective cohort study of non-diabetic OAB patients treated with BTX-A alone or with concurrent GLP-1 therapy. Propensity score matching (1:1) was adjusted for age, race, ethnicity, hypertension, and BMI/obesity. After matching, 992 patients were included in each group. GLP-1 use was associated with a lower incidence of urinary retention (8.6% vs. 4.9%, risk difference 3.66%, p = 0.0044) and urinary tract infection (13.3% vs. 8.8%, risk difference 4.54%, p = 0.00224), with corresponding improved one-year retention-free and UTI-free survival on Kaplan–Meier (KM) analysis. Antispasmodic initiation rates were similar (11.8% vs. 10.3%, risk difference 1.55%, p = 0.6921), and KM analysis showed no significant difference. These findings suggest that GLP-1 receptor agonist use may improve select urinary adverse events in non-diabetic adults undergoing BTX-A treatment for OAB and support further investigation into its potential adjunctive role in OAB management with longer follow-up. Full article

(This article belongs to the Special Issue Botulinum Toxins in Lower-Urinary-Tract Diseases and Dysfunctions)

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14 pages, 1112 KB

Open AccessArticle

Furosemide and the Symptom Burden: The Potential Mediating Role of Uremic Toxins in Patients with CKD

by Margaux Costes-Albrespic, Sophie Liabeuf, Islam-Amine Larabi, Solène M. Laville, Bénédicte Stengel, Abdou Y. Omorou, Luc Frimat, Jean-Claude Alvarez, Ziad A. Massy, Natalia Alencar de Pinho and the CKD-REIN Study Group

Toxins 2025, 17(11), 541; https://doi.org/10.3390/toxins17110541 - 1 Nov 2025

Viewed by 677

Abstract

Furosemide appears to contribute to the accumulation of protein-bound uremic toxins (PBUTs) and to induce adverse drug reactions. We investigated the extent to which the association between the furosemide dose and serum PBUT concentrations mediates the relationship between the furosemide dose and the [...] Read more.

Furosemide appears to contribute to the accumulation of protein-bound uremic toxins (PBUTs) and to induce adverse drug reactions. We investigated the extent to which the association between the furosemide dose and serum PBUT concentrations mediates the relationship between the furosemide dose and the symptom burden in patients with chronic kidney disease (CKD). This cross-sectional analysis included patients with CKD stages 2 to 5 from the CKD-REIN cohort and with data on the baseline serum concentrations of the free fractions of indoxyl sulphate (IS), kynurenine (KYN), p-cresyl sulphate (PCS), and indole-3-acetic acid (IAA), as measured by liquid chromatography–tandem mass spectrometry. The symptom burden was also assessed with a modified (8-item) symptom subscale from the Kidney Disease Quality of Life-36 (e.g., muscle soreness, cramps, itchy skin, dry skin, dizziness, appetite, numbness, and nausea). We used beta regressions to model the association between the furosemide dose and the symptom burden and used structural equation models to quantify the mediating effect of PBUT on this association. Among the 2053 included patients (males: 66%, median age: 68; mean estimated glomerular filtration rate: 35 mL/min/1.73 m²), those prescribed high-dose furosemide (>120 mg/day) had higher symptom burden than those not prescribed furosemide (i.e., a 5.67-point lower symptom score, 95%CI 1.41–9.93). The sum of PBUTs explained 3.78% (95%CI 0.10–18.01%) of this association. Similar results were observed for IS, KYN, and IAA, considered separately, but not for PCS, whose estimated mediation effect was nearly null. Although high-dose furosemide was associated with a greater symptom burden in patients with CKD, mediation by PBUT accumulation appeared to be minimal. Full article

(This article belongs to the Special Issue Uremic Toxins and Hemodialysis: Mechanisms, Challenges, and Therapeutic Advances)

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20 pages, 1479 KB

Open AccessArticle

Functional Effects of BoNT-A Application in Masseter Muscle in Patients with Symptoms of Bruxism

by Krystian Matusz, Artur Drużdż, Natalie Górna, Mariusz Glapiński, Małgorzata Gałczyńska-Rusin, Agata Czajka-Jakubowska, Michał Michalak and Agnieszka Przystańska

Toxins 2025, 17(11), 540; https://doi.org/10.3390/toxins17110540 - 31 Oct 2025

Viewed by 1131

Abstract

Bruxism, defined as a repetitive jaw-muscle activity characterized by clenching or grinding of teeth and/or by bracing or thrusting of the mandible, is a prevalent behavior affecting up to 22% of adults worldwide. While traditionally viewed as a disorder, current understanding recognizes bruxism [...] Read more.

Bruxism, defined as a repetitive jaw-muscle activity characterized by clenching or grinding of teeth and/or by bracing or thrusting of the mandible, is a prevalent behavior affecting up to 22% of adults worldwide. While traditionally viewed as a disorder, current understanding recognizes bruxism as a behavior that may have both positive and negative consequences. Objective assessment methods for evaluating the effectiveness of interventions in symptomatic patients remain limited. This article presents the first longitudinal study using myotonometry to quantify changes in masseter muscle following botulinum toxin type A (BoNT-A) treatment in patients with symptoms of bruxism. In total, 57 patients were recruited and their masseter muscle tone, stiffness, elasticity, relaxation time, and creep parameters were measured. Measurements were performed at baseline, 3 weeks, and 3 months post-injection during both rest and maximum voluntary contraction. BoNT-A treatment produced significant improvements in all biomechanical parameters, with the greatest effects observed in patients with the highest baseline muscle values. The objective biomechanical changes correlated with the duration of BoNT-A’s therapeutic effects. These findings establish myotonometry as a valuable tool for objective assessment of masticatory muscle function and demonstrate that BoNT-A produces measurable, long-lasting biomechanical changes in masseter muscle parameters, supporting its possible clinical application in this challenging condition. Full article

(This article belongs to the Special Issue Botulinum Toxin Therapy for Diseases in the Oral and Maxillofacial Regions)

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11 pages, 2215 KB

Open AccessArticle

Aflatoxigenic Aspergillus Section Flavi Strains and Diverse Molds Isolated from California Almonds

by Barbara Szonyi, Guangwei Huang, Tim Birmingham and Dawit Gizachew

Toxins 2025, 17(11), 539; https://doi.org/10.3390/toxins17110539 - 31 Oct 2025

Viewed by 467

Abstract

Almonds are an essential crop for the economy of California. However, mold and mycotoxin contamination of this commodity has a serious impact on food safety and international trade. The contamination levels of molds and the aflatoxigenic potential of Aspergillus section Flavi isolates were [...] Read more.

Almonds are an essential crop for the economy of California. However, mold and mycotoxin contamination of this commodity has a serious impact on food safety and international trade. The contamination levels of molds and the aflatoxigenic potential of Aspergillus section Flavi isolates were studied on almonds collected at a processing plant in California. The mean total fungal count for 80 samples was 1.0 × 10⁴ CFU/g, while 62 samples (77.5%) had a total mold count less than 1.0 × 10⁴ CFU/g. The most common fungal contaminants were Aspergillus section Nigri (100% of samples), followed by Penicillium (57.5%) and Cladosporium (52.5%) species. Rhizopus, Fusarium and Alternaria spp. were less frequent. A total of 26 A. section Flavi strains were identified, with most strains (23) belonging to the L morphotype of A. flavus. In addition, two S morphotypes of A. flavus, and one A. tamarii strain were observed. Other Aspergillus species, including A. terreus and A. ochraceus were rare. High Performance Liquid Chromatography (HPLC) analysis revealed that 9 out of 13 isolated A. flavus strains produced aflatoxin B₁ (AFB₁) on yeast extract sucrose media. The highest levels of AFB₁ were produced by two A. flavus isolates belonging to the S morphotype (78 and 260 µg/kg). Increasing temperatures and drought conditions may change the population dynamics of toxigenic mold strains on almonds, emphasizing the need to continue monitoring these fungal populations. Full article

(This article belongs to the Special Issue Potential Risks and Analytical Challenges of Mycotoxins in Food and Feed)

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21 pages, 549 KB

Open AccessArticle

Optimisation of a One-Step Reusable Immuno-Affinity Purification Method for the Analysis and Detection of Fumonisin Mycotoxins in Foods and Feeds

by Christian Kosisochukwu Anumudu

Toxins 2025, 17(11), 538; https://doi.org/10.3390/toxins17110538 - 30 Oct 2025

Viewed by 619

Abstract

Fumonisins are among the most prevalent mycotoxins in maize and maize-based products, posing significant food safety and public health risks due to their hepatotoxic, nephrotoxic, and potential carcinogenic effects. Given the strict regulatory limits set by the European Commission and Codex Alimentarius, the [...] Read more.

Fumonisins are among the most prevalent mycotoxins in maize and maize-based products, posing significant food safety and public health risks due to their hepatotoxic, nephrotoxic, and potential carcinogenic effects. Given the strict regulatory limits set by the European Commission and Codex Alimentarius, the development of reliable, sensitive, and matrix–robust analytical methods remain a priority for routine monitoring in both food and feed systems. In this study, a reusable immuno-affinity purification methodology for the quantitative determination of fumonisin mycotoxins (FB1, FB2 and FB3) in foods and feeds (maize matrix) was developed. A single extraction protocol using 2% formic acid in water was employed, followed by cleanup with an immuno-affinity purification column and toxin elution by methanol/PBS (1:1, v/v). Detection and quantification of the mycotoxins was achieved by a normal phase ultra-high performance liquid chromatography coupled with electrospray ionisation triple quadrupole mass spectrometry (UHPLC/ESI-MS/MS). The chromatographic mobile phase utilised was a linear gradient of methanol/water containing 0.1% formic acid. The developed method has a limit of detection of 2.5 ng/g and a limit of quantification of 5 ng/g, all well below the European commission’s guidance values of 1000 ng/g for corn destined for human consumption and 800 ng/g for maize-based breakfast cereals and snacks. While the recovery rates of the method in this study ranged from 65–70% for the three fumonisin analogues in solutions, when tested in maize matrix, recoveries were markedly lower (~30%) due to pronounced matrix suppression. Good repeatability (standard deviation <10%) was achieved for all the fumonisin analogues. The developed method, although quick and effective in solvent systems, suffered limitations to its practical usage due to matrix suppression of the extracts derived from the immuno-affinity purification column, thus significantly reducing the application of the method in measuring fumonisin mycotoxins in food and feed samples. Overall, the method was effective in quantification of fumonisin mycotoxins in solvent solutions but not in food and feed matrices, thus necessitating further optimisation for practical usage. The performance of the developed method was compared to a commercial lateral flow immunochromatographic assay which proved to be better than the developed method in the quantification of toxins in food matrices, as the commercial lateral flow immunochromatographic assay outperformed the developed method in maize matrices. These findings highlight the need for matrix-based validation and further refinement of antibody stability to ensure robust application in regulatory monitoring of fumonisins using immunoaffinity purification methods. Full article

(This article belongs to the Special Issue Mycotoxin Contamination in Animal Nutrition and Its Mitigation Strategies)

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12 pages, 2254 KB

Open AccessArticle

Comprehensive Bibliometric Analysis of Uremic Toxin Research

by Marsuki Hardjo, Takuya Wakamatsu, Kazuki Watanabe, Shoko Yamazaki and Suguru Yamamoto

Toxins 2025, 17(11), 537; https://doi.org/10.3390/toxins17110537 - 30 Oct 2025

Viewed by 1015

Abstract

Uremic toxins accumulate as kidney disease progresses, contributing to diverse systemic disorders. Despite numerous studies, no comprehensive mapping has been performed. In this study, keywords related to uremic toxins were quantitatively analyzed using bibliometric methods to clarify research trends, key molecules, and unresolved [...] Read more.

Uremic toxins accumulate as kidney disease progresses, contributing to diverse systemic disorders. Despite numerous studies, no comprehensive mapping has been performed. In this study, keywords related to uremic toxins were quantitatively analyzed using bibliometric methods to clarify research trends, key molecules, and unresolved challenges. Literature and molecular data on uremic toxins and chronic kidney disease were retrieved from the Web of Science and the European Uremic Toxin Work Group (EUTox) databases. Network, citation burst, and keyword frequency analyses were performed using KeyWords Plus. In total, 3302 articles were identified, showing an increasing trend. Citation burst analysis revealed a growing interest in gut microbiota-related topics (burst strength = 15.21), whereas keyword frequency analysis indicated that indoxyl sulfate (566 articles) and p-cresyl sulfate (537 articles) were the most studied toxins. Toxins such as trimethylamine-N-oxide have gained attention over the past 5 years. Analysis of the EUTox database identified 24 protein-bound uremic toxins; among the 94 toxins with unreported clinical toxicity, 15 molecules, including osteocalcin and quinolinic acid, were investigated in <5 studies. These findings suggest that the gut microbiota and related uremic toxins are current research focuses; however, further investigation of underreported uremic toxins is required to define their clinical significance. Full article

(This article belongs to the Special Issue The Role of Uremic Toxins in Comorbidities of Chronic Kidney Disease)

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15 pages, 2967 KB

Open AccessArticle

Biological Control of Microcystis aeruginosa Through Sequestration in Pseudofaeces Produced by the Freshwater Gastropod, Sinotaia aeruginosa

by Barry N. Madison, Mingzhi Qu, Elliot Gavrin, Wenwei Ren, Yuxiang Wang and Daniel D. Lefebvre

Toxins 2025, 17(11), 536; https://doi.org/10.3390/toxins17110536 - 30 Oct 2025

Viewed by 540

Abstract

Cyanobacteria harmful algal blooms (CHABs) are most commonly caused by the proliferation of the toxic species, Microcystis aeruginosa. It is therefore of considerable interest to develop biological control processes which are economically feasible and scalable for this cyanobacteria that produces the cyanotoxin, [...] Read more.

Cyanobacteria harmful algal blooms (CHABs) are most commonly caused by the proliferation of the toxic species, Microcystis aeruginosa. It is therefore of considerable interest to develop biological control processes which are economically feasible and scalable for this cyanobacteria that produces the cyanotoxin, microcystin. Some gastropods that are abundant in freshwater ecosystems can filter feed on floating planktonic microphytes. We investigated this in the freshwater snail, Sinotaia aeruginosa which indiscriminately accumulated M. aeruginosa, Chlorella vulgaris, and Trichormus variabilis (syn. Anabaena variabilis) The initial filtration rates were approx. 44 and 19 mL · g_wwt⁻¹ · h⁻¹ for unicellular and colony-forming M. aeruginosa, respectively. The pseudofaeces that were formed directly by filtration possessed a limited period of stability, and the bulk of the M. aeruginosa from pseudofaeces was eventually released back to the water column as undigested cyanobacteria. Nevertheless, the rate of sequestration of colonial M. aeruginosa into pseudofaeces was greater than its rate of release, thereby indicating that the temporary stability of pseudofaeces alone would be adequate to impede bloom formation. Therefore, these results provide evidence for using this gastropod in an effective preventative strategy for CHABs formation. Our results highlight the importance of understanding the impact of feeding mechanisms on ecosystem structure when proposing their use in biomanipulative processes aimed at correcting cyanobacteria impacted ecosystems. Full article

(This article belongs to the Section Marine and Freshwater Toxins)

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20 pages, 1877 KB

Open AccessArticle

Photobiomodulation with Combined Wavelengths Results in Improved Clinical Recovery in a Murine Model of Bothrops leucurus Venom Envenomation

by Gisele Dias da Silva, Fabiana Lessa Silva, Anaiá da Paixão Sevá, Juneo Freitas Silva, Danilo Machado Deorce, Nerildo de Jesus da Costa Junior, Fernanda Amaral Silva and Fernando Alzamora Filho

Toxins 2025, 17(11), 535; https://doi.org/10.3390/toxins17110535 - 30 Oct 2025

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Abstract

Snakebite envenomation by Bothrops species is a neglected tropical disease and a major cause of local tissue damage and disability in Latin America. Antivenom therapy is effective against systemic effects but fails to prevent local myonecrosis, inflammation, and pain. This study evaluated photobiomodulation [...] Read more.

Snakebite envenomation by Bothrops species is a neglected tropical disease and a major cause of local tissue damage and disability in Latin America. Antivenom therapy is effective against systemic effects but fails to prevent local myonecrosis, inflammation, and pain. This study evaluated photobiomodulation therapy (PBMT) using infrared (808 nm) alone or in combination with red (660 nm) laser in a murine model of Bothrops leucurus envenomation. A single PBMT session was applied, and animals were evaluated at 24 and 72 h. Combined treatment significantly reduced edema, hyperthermia, plasma CK and LDH, restored nociceptive thresholds, and improved motor recovery compared with infrared alone. Principal component analysis demonstrated clustering of combined-treatment animals with negative controls, supporting a synergistic therapeutic effect. These findings highlight dual-wavelength PBMT as a promising adjunctive approach to antivenom, directly targeting local venom-induced pathology. Full article

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Toxins, Volume 17, Issue 11 (November 2025) – 44 articles

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