Toxins

Article

21 pages, 1306 KB

Open AccessEditor’s ChoiceArticle

Are Baby Rattlesnakes More Dangerous than Adults? Origin, Transmission, and Prevalence of a Media-Driven Myth, with Evidence of Effective Messaging to Dispel It

by William K. Hayes and M. Cale Morris

Toxins 2026, 18(3), 144; https://doi.org/10.3390/toxins18030144 - 14 Mar 2026

Viewed by 9406

Abstract

The easily defanged myth that baby rattlesnakes (genera Crotalus and Sistrurus) are more dangerous than adults has persisted in North America despite all evidence to the contrary. The most often cited reason for the babies-more-dangerous (BMD) myth is the venom-dump (VD) hypothesis: [...] Read more.

The easily defanged myth that baby rattlesnakes (genera Crotalus and Sistrurus) are more dangerous than adults has persisted in North America despite all evidence to the contrary. The most often cited reason for the babies-more-dangerous (BMD) myth is the venom-dump (VD) hypothesis: babies, in contrast to adults, cannot control how much venom they expend, and therefore inject all of it when biting. We undertook three approaches to explore the origin, transmission, and prevalence of this myth and its most frequent explanation. First, we examined historical newspaper accounts. From 130 newspaper stories mentioning the relative danger of baby rattlesnakes, we identified a timeline in which (1) most stories prior to 1969 were factually correct; (2) the BMD myth and VD hypothesis likely originated in the mid-to-late 1960s and became entrenched in California, especially, from 1970 to 1999; (3) factually incorrect statements subsequently prevailed throughout North America from 2000 to 2014; and (4) factually correct stories regained prominence with apparent effective messaging success from 2015 onward. We further learned that general information stories about rattlesnakes, more often citing subject experts like university professors, were much more likely to provide accurate information than local snakebite stories, which more often cited health professionals (e.g., physicians, veterinarians, pharmacists) and emergency responders (e.g., police and fire officers) who frequently supplied misinformation. Second, we surveyed familiarity with the BMD myth and VD hypothesis among 53 university classrooms (including one high school) representing 3751 students across 29 states within the United States. Consistent with the California media’s outsized influence on misinformation transmission, familiarity with the myth was greatest in the southwestern states (52.6%) and declined moving north and east, with the least familiarity in the northeastern states (16.4%). Third, a small survey of 75 emergency responders and health professionals from Southern California revealed that a whopping 73.3% actually believed the BMD myth. Numerous organizations generally regarded as authoritative further amplified the misinformation, especially on the internet, where some content persists to this day. Unfortunately, belief in the BMD myth and VD hypothesis can lead to negative consequences, including misinformed risk-taking by those encountering snakes, unwarranted fear among snakebite victims, and inappropriate care delivered by misinformed or patient/family-pressured medical professionals. Our findings target health professionals and emergency responders as priority audiences for education. Full article

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19 pages, 2381 KB

Open AccessEditor’s ChoiceArticle

RTP004 Peptide Binds to Botulinum Neurotoxin, Increases Cell Surface Binding, and Enhances Cellular SNAP-25 Cleavage

by Andre F. Batista, Ratnesh Singh, Frank Lee, Shaoqiu Zhuo, Dmitri Leonoudakis and Conor J. Gallagher

Toxins 2026, 18(3), 134; https://doi.org/10.3390/toxins18030134 - 10 Mar 2026

Viewed by 1826

Abstract

DaxibotulinumtoxinA for injection (DAXI) is a botulinum neurotoxin (BoNT) drug product comprising the 150 kDa pure BoNT/A1 as the drug substance formulated with a proprietary stabilizing excipient, RTP004. We hypothesized that RTP004 facilitates localization of BoNT/A1 to the neuronal membrane, resulting in increased [...] Read more.

DaxibotulinumtoxinA for injection (DAXI) is a botulinum neurotoxin (BoNT) drug product comprising the 150 kDa pure BoNT/A1 as the drug substance formulated with a proprietary stabilizing excipient, RTP004. We hypothesized that RTP004 facilitates localization of BoNT/A1 to the neuronal membrane, resulting in increased BoNT internalization and cleavage of the synaptosomal-associated protein of 25 kDa (SNAP-25) within synaptic terminals. We characterized the interaction between RTP004 and BoNT/A1 using in silico and in vitro techniques. In vitro analyses revealed that negative charges on the BoNT/A1 surface were located on the light chain (LC, the catalytic domain) and the C-terminus of the heavy chain (H_C, the receptor-binding domain), potentially providing sites for interaction with the positively charged RTP004 peptide. RTP004 bound to BoNT/A1, but not to human serum albumin (HSA), in both static and dynamic conditions. RTP004, not HSA, enhanced binding of BoNT to artificial membranes and RTP004 dissociated from BoNT under conditions that mimicked physiological conditions of the synaptic vesicle. RTP004 also increased binding of BoNT to the synaptosomal cell membrane and enhanced cleavage of SNAP-25 in a dose-dependent manner. These findings demonstrate that RTP004, not the excipient HSA common in other BoNT/A1 drug products, enhances binding of BoNT to the cell surface, facilitates internalization of BoNT into the cell, and increases SNAP-25 cleavage. Full article

(This article belongs to the Section Bacterial Toxins)

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20 pages, 4381 KB

Open AccessEditor’s ChoiceArticle

Influence of Plant Age and Endophyte Status on the Nematotoxicity of Festulolium loliaceum to Trichodorus primitivus and Quantification of Active Phytochemicals

by Nyambura G. Mwangi, Timothy J. Gillanders, Mark Stevens, Alistair J. D. Wright, Simon G. Edwards, Martin C. Hare and Matthew A. Back

Toxins 2026, 18(3), 125; https://doi.org/10.3390/toxins18030125 - 1 Mar 2026

Viewed by 1554

Abstract

Festulolium hybrids are cool-season forage grasses that form symbiotic relationships with the fungus Epichloë uncinata, which produces loline alkaloids that protect the host from herbivores. This study evaluated the nematotoxicity of shoot and root extracts of Festulolium loliaceum against the stubby root [...] Read more.

Festulolium hybrids are cool-season forage grasses that form symbiotic relationships with the fungus Epichloë uncinata, which produces loline alkaloids that protect the host from herbivores. This study evaluated the nematotoxicity of shoot and root extracts of Festulolium loliaceum against the stubby root nematode Trichodorus primitivus. Methanolic root and shoot extracts from plants aged 8, 12, 16, and 20 weeks were tested in vitro at five concentrations (312.5–5000 µg mL⁻¹) over 24, 48, and 72 h. Nematode immobility, mortality, and phytochemical profiles, including flavonoids, loline alkaloids, and phenols, were quantified. Extracts from shoots caused significant concentration and time-dependent immobility of T. primitivus (p = 0.001), reaching ≥90% at 5000 µg mL⁻¹ after 72 h in 8–12-week-old plants. Endophyte presence enhanced nematotoxicity, where LD₅₀ values for E+ roots were two-fold lower at 12 weeks and fifty-fold lower at 20 weeks compared with E− root extracts. Shoot extracts of E+ grass had the highest nematicidal activity at 8 weeks, with a significantly lower LD₅₀ value than E− (p < 0.05). Loline alkaloid concentrations increased with plant age, while flavonoids and phenols declined. Nematotoxicity of F. loliaceum extracts was strongly influenced by plant age and endophyte presence. Younger E+ shoots produced the most potent shoot extracts, whereas older plants produced the most potent root extracts. Flavonoid content was negatively correlated with shoot biomass (R = −0.94, p < 0.001). Similarly, phenol content was negatively correlated to both root biomass (R = −0.79, p < 0.001) and shoot biomass (R = −0.67, p < 0.005). Full article

(This article belongs to the Section Plant Toxins)

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18 pages, 2032 KB

Open AccessEditor’s ChoiceArticle

Inter- and Intraspecific Venom Variation in the Reclusive Rear-Fanged Black-Striped Snakes (Coniophanes)

by John Henry Fowler, Ramses Alejandro Rosales-García, Rhett M. Rautsaw, Michael P. Hogan, Erich P. Hofmann, Andrew J. Mason, Ramon Nagesan, Miguel Borja, Luis Herrera, Gamaliel Castañeda-Gaytan, Alison R. Davis Rabosky, Darin R. Rokyta and Christopher L. Parkinson

Toxins 2026, 18(2), 108; https://doi.org/10.3390/toxins18020108 - 20 Feb 2026

Viewed by 2487

Abstract

Our current understanding of snake venom is highly biased towards species known to be medically significant in human envenomations. This vastly under-represents the true evolutionary and ecological breadth of snake venom, with gaps spanning entire clades and unique lifestyles. As a result, many [...] Read more.

Our current understanding of snake venom is highly biased towards species known to be medically significant in human envenomations. This vastly under-represents the true evolutionary and ecological breadth of snake venom, with gaps spanning entire clades and unique lifestyles. As a result, many genera of rear-fanged snakes lack well-understood venom profiles despite these taxa composing around 65% of known extant snake species. Methodological challenges associated with venom extraction have long been a key reason responsible for the lack of venom research on this group. Modern advancements in venomics technologies have allowed researchers to overcome many of these challenges and investigate the venom components of understudied genera. The genus Coniophanes (black-striped snakes) presents an ideal system for investigating venom and the venom delivery system in a rear-fanged venomous species with well-documented accounts of human envenomations. We sequenced and annotated de novo transcriptomes of the Duvernoy’s gland (DVG) for seven individuals across four species of Coniophanes (Dipsadidae) and confirmed toxin expression in representative venom proteomes. We assessed interspecific venom variation within this genus and further examined intraspecific venom variation within C. imperialis. We found that toxins account for 38.8% to 66% of the total DVG transcriptomes and that 18 toxin families are represented in this genus, with prominent expression of cystine-rich secretory proteins (CRiSPs) in three species and snake venom metalloproteinases (SVMPs) in all four species. In addition, we used diffusible iodine-based contrast-enhanced computed tomography (diceCT) to better understand the venom delivery system for C. fissidens, a widespread species within this genus, showcasing enlarged, grooved, rear fangs in close proximity to a prominent DVG. We provide the first ever characterization of the venom profiles of Coniophanes, highlight venom variation between and within species, and outline the venom delivery system of this understudied genus. Full article

(This article belongs to the Special Issue Snake Venom Genes Expression, Evolution and Variation)

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11 pages, 1135 KB

Open AccessEditor’s ChoiceArticle

Skin Irritation-Associated Dinoflagellate Vulcanodinium rugosum Isolated from Cienfuegos Bay, Cuba: Toxin Profile and Cell Growth Characterization Under Laboratory Conditions

by Angel R. Moreira-Gonzalez, Catarina Churro, Vera Marques, Lisbet Díaz-Asencio, Donaida Chamero Lago and Pedro Reis Costa

Toxins 2026, 18(2), 96; https://doi.org/10.3390/toxins18020096 - 14 Feb 2026

Viewed by 2189

Abstract

Blooms of the marine dinoflagellate Vulcanodinium rugosum have been associated with skin lesion outbreaks in Cuba and elsewhere. In this study, cell growth and toxin production were investigated under laboratory-controlled conditions in two strains isolated from Cienfuegos Bay, Cuba. Strains were cultured with [...] Read more.

Blooms of the marine dinoflagellate Vulcanodinium rugosum have been associated with skin lesion outbreaks in Cuba and elsewhere. In this study, cell growth and toxin production were investigated under laboratory-controlled conditions in two strains isolated from Cienfuegos Bay, Cuba. Strains were cultured with and without a mechanical agitation and toxins were analyzed at two stages of the culture growth (exponential and stationary). Although blooms in Cienfuegos Bay occur in a semi-enclosed system characterized by calm waters with no agitation, the results of this study suggest that V. rugosum cells may also exhibit growth capacity under agitated conditions, or in open waters, comparable to that observed in systems with low hydrodynamic energy. Higher toxin levels, as determined by liquid chromatography with mass spectrometry (LC-MS/MS), were detected after exponential growth. Portimine-A and pinnatoxin-F (PnTX-F) were the dominant toxins (up to 1.75 and 1.0 pg·cell⁻¹, respectively). PnTX-E, -D and Portimine-B were also detected at minor concentrations. This study contributes the first data necessary for a proper interpretation of monitoring programs aiming to assess the impact of V. rugosum blooms, particularly when used alongside forecasting models. Full article

(This article belongs to the Special Issue Unveiling the Toxic Effects of Harmful Algal Blooms: 2nd Edition)

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28 pages, 1112 KB

Open AccessEditor’s ChoiceArticle

Microcystin-LR Drives Early NAFLD Pathogenesis via Hepatic Cholesterol Accumulation: Dysregulation of Ldlr and Abcg1 Expression Uncoupled from Srebp2

by Hideaki Kawahara, Yoshihito Koto, Yuuka Hitsuda, Koichi Kurata, Keisuke Yoshikiyo, Ayumi Hashiguchi, Hideaki Maseda, Kunihiro Okano, Norio Sugiura, Kazuya Shimizu and Hidehisa Shimizu

Toxins 2026, 18(2), 92; https://doi.org/10.3390/toxins18020092 - 11 Feb 2026

Cited by 1 | Viewed by 1529

Abstract

Chronic exposure to the cyanotoxin microcystin-LR is an emerging environmental driver of non-alcoholic fatty liver disease (NAFLD); however, the initiating molecular events at sub-lethal, environmentally relevant concentrations remain elusive. Current safety guidelines focus primarily on acute injury, potentially overlooking silent metabolic disruption. The [...] Read more.

Chronic exposure to the cyanotoxin microcystin-LR is an emerging environmental driver of non-alcoholic fatty liver disease (NAFLD); however, the initiating molecular events at sub-lethal, environmentally relevant concentrations remain elusive. Current safety guidelines focus primarily on acute injury, potentially overlooking silent metabolic disruption. The present study investigates the early metabolic toxicity of chronic low-dose microcystin-LR (10 µg/L) in a 7-week rat model, specifically focusing on pre-symptomatic perturbations in lipid homeostasis. By integrating biochemical profiling with multivariate systems toxicology (LASSO and PLS-DA), we identified a specific phenotype of “Silent Hepatic Total Cholesterol Accumulation” (T-CHOL +16%, p = 0.01) occurring in the absence of systemic dyslipidemia or overt liver injury. Mechanistic analysis revealed a specific dual failure of cholesterol homeostasis, characterized by the paradoxical upregulation of the influx transporter Ldlr (LASSO coef +0.661) and the suppression of the efflux transporter Abcg1 (PLS1 loading −0.358). Crucially, Ldlr upregulation occurred despite the concomitant transcriptional downregulation of Srebp2 (Spearman ρ = −0.585), indicating a regulatory uncoupling mechanism. We propose that microcystin-LR-induced protein phosphatase 2A (PP2A) inhibition likely drives this uncoupling via a post-transcriptional override—possibly involving ERK/RSK-mediated Ldlr mRNA stabilization. Concurrently, this inhibition appears to block LXR-mediated Abcg1 expression through sustained AMPK hyperactivation resulting from the loss of dephosphorylation. These findings indicate liver-specific cholesterol accumulation as the critical first step of environmental NAFLD pathogenesis, suggesting that current WHO guidelines (1 µg/L) may require re-evaluation regarding metabolic safety. We propose the hepatic Ldlr/Abcg1 ratio as a potential early biomarker for revised risk assessment. Full article

(This article belongs to the Special Issue Advances in Detecting, Monitoring, Predicting, Managing and Controlling Harmful Algal Blooms)

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21 pages, 2443 KB

Open AccessEditor’s ChoiceArticle

Mycotoxins and Beyond: Unveiling Multiple Organic Contaminants in Pet Feeds Through HRMS Suspect Screening

by Dafni Dematati, Dimitrios Gkountouras, Vasiliki Boti and Triantafyllos Albanis

Toxins 2026, 18(1), 22; https://doi.org/10.3390/toxins18010022 - 30 Dec 2025

Viewed by 7216

Abstract

This study evaluated 65 commercially available pet feed samples, including 33 cat feeds and 32 dog feeds (dry and wet formulations), for the presence of organic contaminants. These included mycotoxins, pesticides, pharmaceutical residues/veterinary drugs, and plant-based bioactive compounds. A suspect screening strategy was [...] Read more.

This study evaluated 65 commercially available pet feed samples, including 33 cat feeds and 32 dog feeds (dry and wet formulations), for the presence of organic contaminants. These included mycotoxins, pesticides, pharmaceutical residues/veterinary drugs, and plant-based bioactive compounds. A suspect screening strategy was employed using QuEChERS extraction followed by LC-LTQ/Orbitrap HRMS analysis. A total of 29 compounds were tentatively identified within 186 detections. In total, 76.9% of the samples were contaminated with mycotoxins. Aflatoxins (B1, B2, G1, and G2), T2 toxins, and HT2 toxins were dominant, with Aflatoxin B1 occurring in 33.8% of the samples and exhibiting a higher prevalence in dry feeds than in wet feeds. Pesticides were present in 72.0% of the dry formulations, including aclonifen and pirimiphos-methyl, but were present in only 11% of the wet formulations. Plant-based bioactive compounds, including phytoestrogens, were identified in 51% of the samples, highlighting toxicologically relevant candidates that merit prioritization for targeted confirmation, particularly in cat feeds. Pharmaceuticals were found in 23.8% of dry feeds (sparfloxacin and fumagillin). Overall, the HRMS-based, standard-free suspect screening workflow provides an early-warning overview of multi-class co-occurrence patterns in complex pet feed matrices and supports the prioritization of candidates for subsequent confirmatory analysis. Full article

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

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27 pages, 3378 KB

Open AccessEditor’s ChoiceArticle

Probiotic Supplementation in Chronic Kidney Disease: Outcomes on Uremic Toxins, Inflammation, and Vascular Calcification from Experimental and Clinical Models

by Teresa Obrero, María Victoria Pendón-Ruiz de Mier, Jose E. Gordillo-Arnaud, María José Jiménez Moral, Victoria Vidal, Fátima Guerrero, Andrés Carmona, María Encarnación Rodríguez-Ortiz, Ana Isabel Torralbo, Raquel Ojeda, Cayetana Moyano, Mercedes Sanchez-Ramade, Juan Mesa, Daniel J. López-Ruiz, Karen Valdés-Díaz, Raquel María García-Sáez, Daniel Jurado-Montoya, Cristian Rodelo-Haad, María Álvarez-Benito, Griet Glorieux, Mariano Rodríguez, Sagrario Soriano-Cabrera and Juan Rafael Muñoz-Castañeda Show full author list Hide full author list

Toxins 2026, 18(1), 6; https://doi.org/10.3390/toxins18010006 - 20 Dec 2025

Cited by 2 | Viewed by 3292

Abstract

Chronic kidney disease (CKD) is associated with gut microbiota alterations that contribute to increased inflammation and the generation of uremic toxins and may worsen the disease progression. While probiotics may improve the pro-inflammatory cytokine profile, their effects on mineral metabolism, vascular calcification (VC), [...] Read more.

Chronic kidney disease (CKD) is associated with gut microbiota alterations that contribute to increased inflammation and the generation of uremic toxins and may worsen the disease progression. While probiotics may improve the pro-inflammatory cytokine profile, their effects on mineral metabolism, vascular calcification (VC), and CKD progression remain unclear. We aimed to evaluate the impact of a commercial probiotic (Probimel) supplementation on kidney function, mineral metabolism, inflammation and VC in both an experimental rat model and patients with advanced CKD and VC. The experimental model of VC was performed through 5/6 nephrectomy (Nx), a high-phosphate diet, and calcitriol. Animals were divided into three groups: Sham, Nephrectomy, and Nephrectomy + Probiotic. In the exploratory clinical study, 23 patients with advanced stage 5 CKD and VC were randomized and either received or did not receive daily probiotics for 6 months. Kidney function, mineral metabolism, uremic toxins, inflammation, VC, and fecal microbiota were evaluated. Probiotic supplementation decreased interleukin-6 (IL-6) and interpheron-γ (IFN-γ) and levels of the uremic toxin, indoxyl sulfate (IS), in the experimental model. However, no clear evidence of improvement in kidney function or vascular calcification was observed in either rats or patients with this probiotic. Under our experimental and clinical conditions, the selected probiotic did not modify key parameters related to CKD progression or VC. Full article

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

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19 pages, 2956 KB

Open AccessEditor’s ChoiceArticle

Ultrasound Evaluation of Upper Facial Muscles to Guide Botulinum Toxin Application

by Dominika Jaguś, Anna Pawłowska and Robert Krzysztof Mlosek

Toxins 2025, 17(12), 595; https://doi.org/10.3390/toxins17120595 - 14 Dec 2025

Cited by 4 | Viewed by 1602

Abstract

Background: Botulinum toxin injection is one of the most common esthetic procedures, yet complications may occur due to anatomical variability or suboptimal injection technique. This study aimed to evaluate the upper facial muscles using ultrasound, focusing on inter- and intraindividual variability. Methods: The [...] Read more.

Background: Botulinum toxin injection is one of the most common esthetic procedures, yet complications may occur due to anatomical variability or suboptimal injection technique. This study aimed to evaluate the upper facial muscles using ultrasound, focusing on inter- and intraindividual variability. Methods: The study involved volunteers aged 21–40 years, excluding those with prior facial treatments, trauma, or muscle disorders. The muscles examined included the occipitofrontalis (frontal belly), procerus, corrugator supercilii, and orbicularis oculi. Muscle thickness and distance from the epidermis were measured using high-frequency ultrasound. Statistical analyses included descriptive statistics, correlation with age and BMI, sex comparisons, and symmetry assessment. Results: A total of 127 participants (103 women and 24 men) were enrolled, with a mean age of 28.8 ± 4.4 years. Age showed no significant correlation with muscle thickness or depth, supporting the internal consistency of the studied age group. BMI showed moderate correlations with the depth of the selected forehead muscles. Males showed greater thickness in the frontal and procerus muscles. Relative side-to-side asymmetry coefficients reached 40% for both thickness and depth, indicating notable individual laterality. Conclusions: The study provides normative ultrasound parameters for the upper facial muscle in healthy adults. The results demonstrate significant anatomical variability depending on sex, BMI, and facial laterality, supporting individualized ultrasound-guided approaches for botulinum toxin injection. Full article

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18 pages, 2187 KB

Open AccessEditor’s ChoiceArticle

Acute Amanita muscaria Toxicity: A Literature Review and Two Case Reports in Elderly Spouses Following Home Preparation

by Stanila Stoeva-Grigorova, Ivanesa Yarabanova, Maya Radeva-Ilieva, Diana Ivanova, Snezha Zlateva and Petko Marinov

Toxins 2025, 17(12), 570; https://doi.org/10.3390/toxins17120570 - 25 Nov 2025

Cited by 5 | Viewed by 12093

Abstract

Amanita muscaria (L.) Lam., commonly known as fly agaric, remains an uncommon yet clinically important cause of acute mushroom intoxication. Although typically associated with mild to moderate neuropsychiatric disturbances, the mushroom’s toxic profile is highly variable and continues to attract scientific, toxicological, [...] Read more.

Amanita muscaria (L.) Lam., commonly known as fly agaric, remains an uncommon yet clinically important cause of acute mushroom intoxication. Although typically associated with mild to moderate neuropsychiatric disturbances, the mushroom’s toxic profile is highly variable and continues to attract scientific, toxicological, and public health interest. This work provides an integrative review of the biochemical composition, toxicodynamics, and clinical manifestations associated with A. muscaria exposure, with particular emphasis on the pharmacological actions of its principal constituents, ibotenic acid and muscimol. The review is complemented by two contemporaneous cases of severe intoxication in elderly individuals, illustrating the real-world clinical expression of the toxidrome and the challenges in diagnosis and management. Both cases presented with rapid-onset gastrointestinal symptoms, profound central nervous system depression, and cholinergic features, requiring intensive supportive therapy, atropine infusion, and continuous monitoring. Full recovery was achieved in both patients. These clinical observations contextualize the broader toxicological framework discussed in the review and underscore the need for increased clinical vigilance, improved public education, and strengthened diagnostic and therapeutic preparedness regarding psychoactive wild mushroom exposures. Full article

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12 pages, 438 KB

Open AccessEditor’s ChoiceArticle

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

Cited by 5 | Viewed by 2536

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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18 pages, 553 KB

Open AccessEditor’s ChoiceArticle

AI and Biotechnology to Combat Aflatoxins: Future Directions for Modern Technologies in Reducing Aflatoxin Risk

by Charitha J. Gamlath and Felicia Wu

Toxins 2025, 17(11), 524; https://doi.org/10.3390/toxins17110524 - 23 Oct 2025

Cited by 5 | Viewed by 2161

Abstract

Although a decades-old problem in food safety, aflatoxin has largely resisted human control methods. This situation could be mitigated using new technologies that could provide better control all along the food supply chain, for crops frequently infected with the causative fungi Aspergillus flavus [...] Read more.

Although a decades-old problem in food safety, aflatoxin has largely resisted human control methods. This situation could be mitigated using new technologies that could provide better control all along the food supply chain, for crops frequently infected with the causative fungi Aspergillus flavus and A. parasiticus, which produce aflatoxin. Generative artificial intelligence (AI) and modern biotechnology could offer, and have offered, a suite of potential solutions to reducing both fungal infection and aflatoxin contamination of foods. In this paper, we describe these technologies, as well as means by which they may be utilized to reduce aflatoxin risk along the food supply chain. We discuss how regulatory frameworks worldwide may be restrictive for biotechnologies in certain parts of the world, but are relatively less stringent for AI at present. To the extent that these technologies can be harnessed and deployed safely to combat the problem of aflatoxins, we encourage research and development in these areas to improve the precision, accuracy, and speed by which to deal with this food safety risk. Full article

(This article belongs to the Special Issue 65 Years On from Aflatoxin Discovery—a Themed Issue in Honor of Professor John D. Groopman)

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21 pages, 805 KB

Open AccessEditor’s ChoiceArticle

Unilateral EMG-Guided Botulinum Toxin for Retrograde Cricopharyngeus Dysfunction: A Prospective Clinical and Neurophysiological Study

by Giuseppe Cosentino, Chiara Zaffina, Clara Zoccola, Mauro Fresia, Sara Merli, Simone Mauramati, Giulia Bertino, Massimiliano Todisco, Shayan Dodge, Sami Barmada, Enrico Alfonsi and Cristina Tassorelli

Toxins 2025, 17(9), 458; https://doi.org/10.3390/toxins17090458 - 12 Sep 2025

Cited by 5 | Viewed by 2877

Abstract

Retrograde cricopharyngeus dysfunction (R-CPD) is a recently recognized condition characterized by the inability to burp, typically accompanied by gurgling noises, bloating, and flatulence. Percutaneous botulinum neurotoxin (BoNT) injection into the cricopharyngeus muscle is a minimally invasive treatment with promising effects, although current evidence [...] Read more.

Retrograde cricopharyngeus dysfunction (R-CPD) is a recently recognized condition characterized by the inability to burp, typically accompanied by gurgling noises, bloating, and flatulence. Percutaneous botulinum neurotoxin (BoNT) injection into the cricopharyngeus muscle is a minimally invasive treatment with promising effects, although current evidence remains limited. In this prospective, open-label study, we evaluated the clinical effects of increasing doses (10 to 30 U) of EMG-guided unilateral BoNT injection in 67 patients with R-CPD. Symptom severity and quality of life were assessed at baseline and at 1 and 4 months post-treatment. The electromyographic (EMG) parameters of the cricopharyngeus were recorded to explore their association with symptom burden and treatment response. At a 1-month follow-up, 55.2% of patients were classified as responders (satisfaction score ≥ 6/10), with a higher rate (64.4%) observed at higher doses, particularly in female patients. Both symptom severity and quality of life improved significantly at 1 month and were sustained at 4 months. Higher cricopharyngeus EMG activity was associated with more severe symptoms and lesser treatment responses. Unilateral EMG-guided BoNT injection is a safe and effective treatment for R-CPD. Further studies should explore the potential role of electromyography in clarifying the pathophysiological aspects of R-CPD and guiding treatment. Full article

(This article belongs to the Special Issue Botulinum Toxins: New Uses in the Treatment of Diseases (2nd Edition))

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18 pages, 4803 KB

Open AccessEditor’s ChoiceArticle

Exploring the Potential of Genista ulicina Phytochemicals as Natural Biocontrol Agents: A Comparative In Vitro and In Silico Analysis

by Roukia Zatout, Ouided Benslama, Fatima Zohra Makhlouf, Alessio Cimmino, Maria Michela Salvatore, Anna Andolfi, Radhia Manel Kolla and Marco Masi

Toxins 2025, 17(9), 452; https://doi.org/10.3390/toxins17090452 - 6 Sep 2025

Cited by 9 | Viewed by 1521

Abstract

Development of new sustainable pesticides represents a real challenge for researchers due to environmental issues and public health aspects. In fact, the overuse of chemical pesticides has led to environmental damage, loss of biodiversity, and pesticide-resistant pests. In a framework characterized by the [...] Read more.

Development of new sustainable pesticides represents a real challenge for researchers due to environmental issues and public health aspects. In fact, the overuse of chemical pesticides has led to environmental damage, loss of biodiversity, and pesticide-resistant pests. In a framework characterized by the necessity of new sustainable agricultural practices, this study investigates the plant Genista ulicina as a producer of bioactive compounds for potential application as eco-friendly biopesticides. First, both roots and aerial parts of G. ulicina were extracted and the main compounds in the crude extracts were identified via GC-MS. Subsequently, the crude extracts were submitted to antifungal and phytotoxic assays. In particular, the antifungal effects were evaluated on three common phytopathogenic fungi, Fusarium oxysporum, Alternaria alternata, and Botrytis cinerea, while phytotoxic activity was evaluated on two weed species: Euphorbia peplus L. and Oxalis corniculata L. Further insights were obtained on the herbicidal potential of phytochemical compounds produced by G. ulicina through in silico investigations. In particular, molecular docking analyses were performed against three key enzymes involved in essential plant metabolic pathways: acetohydroxyacid synthase (AHAS), 4-hydroxyphenylpyruvate dioxygenase (HPPD), and protoporphyrinogen oxidase (PPO). Among the compounds identified, linolelaidic acid methyl ester, 1-monolinolein, stearic acid, and palmitic acid derivatives showed promising binding affinities and favorable interaction patterns compared to reference ligands. Selected phytochemicals from G. ulicina show potential as inhibitors of key herbicide targets, suggesting their value as promising leads in the development of sustainable bio-based weed control agents. Full article

(This article belongs to the Section Plant Toxins)

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23 pages, 2745 KB

Open AccessEditor’s ChoiceArticle

Pioneering Comparative Proteomic and Enzymatic Profiling of Amazonian Scorpion Venoms Enables the Isolation of Their First α-Ktx, Metalloprotease, and Phospholipase A₂

by Karla C. F. Bordon, Gabrielle C. Santos, Jonas G. Martins, Gisele A. Wiezel, Fernanda G. Amorim, Thomas Crasset, Damien Redureau, Loïc Quinton, Rudi E. L. Procópio and Eliane C. Arantes

Toxins 2025, 17(8), 411; https://doi.org/10.3390/toxins17080411 - 15 Aug 2025

Cited by 8 | Viewed by 3356

Abstract

Scorpionism is a growing public health concern in Brazil, with the Amazon region presenting the highest mortality rates but remaining understudied, especially regarding local scorpion venoms composition. This study presents the first comprehensive biochemical characterization of venoms from three Amazonian species—Tityus metuendus [...] Read more.

Scorpionism is a growing public health concern in Brazil, with the Amazon region presenting the highest mortality rates but remaining understudied, especially regarding local scorpion venoms composition. This study presents the first comprehensive biochemical characterization of venoms from three Amazonian species—Tityus metuendus (TmetuV), Tityus silvestris (TsilvV), and Brotheas amazonicus (BamazV)—using an integrated approach combining Multi-Enzymatic Limited Digestion (MELD)-based bottom-up proteomics, high-resolution LC-MS/MS, chromatography, zymography, and enzymatic assays. Tityus serrulatus venom was included as a reference. Significant biochemical differences were observed: TsilvV was rich in 20–30 kDa proteins and showed strong metalloprotease activity; BamazV exhibited high molecular weight proteins and potent phospholipase A₂ (PLA₂) activity but lacked proteolytic and fibrinogenolytic activities; TmetuV showed the highest hyaluronidase activity and abundance of α-KTx neurotoxins. Zymography revealed a conserved ~45 kDa hyaluronidase in all species. Three novel components were partially characterized: BamazPLA₂ (Group III PLA₂), Tmetu1 (37-residue α-KTx), and TsilvMP_A (a metalloprotease homologous to antarease). This is the first application of MELD-based proteomics to Amazonian scorpion venoms, revealing molecular diversity and functional divergence within Tityus and Brotheas, emphasizing the need for region-specific antivenoms. These findings provide a foundation for future pharmacological studies and the discovery of bioactive peptides with therapeutic potential. Full article

(This article belongs to the Special Issue Unlocking the Deep Secrets of Toxins)

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16 pages, 574 KB

Open AccessEditor’s ChoiceArticle

In Vitro Evaluation of Aflatoxin B1 Detoxification by Lactobacillus, Pediococcus, and Bacillus Strains

by Sarra Rafai, Ana Moreno, Alessandra Cimbalo, Pilar Vila-Donat, Lara Manyes and Giuseppe Meca

Toxins 2025, 17(8), 403; https://doi.org/10.3390/toxins17080403 - 11 Aug 2025

Cited by 10 | Viewed by 3128

Abstract

Biologically based detoxification strategies are increasingly being explored as alternatives to conventional methods for the removal of toxic contaminants in food products. Among these, aflatoxin B1 (AFB1) is one of the most potent mycotoxins due to its high toxicity, genotoxicity, and persistence in [...] Read more.

Biologically based detoxification strategies are increasingly being explored as alternatives to conventional methods for the removal of toxic contaminants in food products. Among these, aflatoxin B1 (AFB1) is one of the most potent mycotoxins due to its high toxicity, genotoxicity, and persistence in the human body once ingested. In this study, the detoxification potential of bacterial strains belonging to the genera Lactobacillus/Pediococcus (n = 10) and Bacillus (n = 10) was evaluated using extracts from naturally contaminated corn flour. Detoxification was assessed after incubation for 12, 24, and 48 h in specific culture media. AFB1 quantification and metabolite profiling were performed at each time point using Quadrupole Time-of-Flight Mass Spectrometry (LC-QTOF-MS). The highest detoxification rates were observed with Lactobacillus curvatus 14 (L. curvatus 14) (41.1 ± 19.3%) and Pediococcus pentosaceus 4 (P. pentosaceus 4) (25.4 ± 11.3%) after 48 h, and Bacillus firmus 6 (B. firmus 6) (25.1 ± 12.9%) after 24 h. An in vitro digestion model was also applied to assess detoxification under gastrointestinal conditions. Results showed substantial AFB1 reduction at the colonic stage, reaching 72.26 ± 7.54% for P. pentosaceus 4 and 69.67 ± 9.70% for L. curvatus 14. These findings underscore the potential application of Lactobacillus, Pediococcus, and Bacillus strains in biological detoxification strategies to reduce dietary exposure to AFB1. Full article

(This article belongs to the Special Issue Aflatoxins: Contamination, Analysis and Control)

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15 pages, 2398 KB

Open AccessEditor’s ChoiceArticle

Heating up the Blunts: Prothrombin Activation, with Factor Va as an Obligate Cofactor, Is the Dominant Procoagulant Mechanism of Blunt-Nosed Viper Venoms (Macrovipera Species)

by Patrick S. Champagne, Lorenzo Seneci and Bryan G. Fry

Toxins 2025, 17(8), 398; https://doi.org/10.3390/toxins17080398 - 8 Aug 2025

Cited by 7 | Viewed by 3748

Abstract

Venoms of the Palearctic vipers in the Macrovipera genus cause severe procoagulant clinical effects, yet the precise molecular targets remain incompletely defined. To fill this toxicological knowledge gap, we tested five Macrovipera venoms—M. lebetina cernovi, M. l. obtusa, M. l. [...] Read more.

Venoms of the Palearctic vipers in the Macrovipera genus cause severe procoagulant clinical effects, yet the precise molecular targets remain incompletely defined. To fill this toxicological knowledge gap, we tested five Macrovipera venoms—M. lebetina cernovi, M. l. obtusa, M. l. turanica (Turkmenistan and Uzbekistan localities), and M. schweizeri—using plasma clotting assays, Factors VII, X, XI, and XII and prothrombin zymogen activation assays, and SDS-PAGE to visualise Factor V (FV) cleavage. All venoms induced extremely rapid clot formation (10.5–12.5 s) compared with the negative control (spontaneous clotting) of 334.6 ± 3.6 s) and the positive control (kaolin trigger) of 55.8 ± 1.9 s. Activation of FVII or FXI was negligible, whereas consistent FX activation and species-variable FXII activation, both moderate, were observed. Prothrombin remained inert in the absence of cofactors, but the presence of FV or FVa elicited potent thrombin generation. SDS-PAGE confirmed proteolytic conversion of the 330 kDa FV zymogen into the ~105 kDa heavy and ~80 kDa light chains of FVa by the venoms of all species. This data demonstrates that Macrovipera venoms rely on a dual enzyme strategy: (i) activation of FV to FVa by serine proteases and (ii) FVa-dependent prothrombin activation by metalloproteases. These results reveal that prothrombin activation is the dominant procoagulant pathway and overshadows the historically emphasised FX activation. This mechanism mirrors, yet is evolutionarily independent from, the FXa:FVa prothrombinase formation seen in Australian elapid venoms, highlighting convergent evolution of cofactor-hijacking strategies among snakes. The discovery of potent FVa-mediated prothrombin activation in Macrovipera challenges existing paradigms of viperid venom action, prompts re-evaluation of related genera (e.g., Daboia), and underpins the design of targeted antivenom and therapeutic interventions. Full article

(This article belongs to the Special Issue Toxins from Venoms and Poisons)

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14 pages, 510 KB

Open AccessEditor’s ChoiceArticle

Is There a “Non-Motor Effect” of Botulinum Toxin Treatment in Cervical Dystonia in Addition to Its Effects on Motor Symptoms?

by Małgorzata Dudzic, Anna Pieczyńska, Artur Drużdż, Anna Rajewska and Katarzyna Hojan

Toxins 2025, 17(8), 396; https://doi.org/10.3390/toxins17080396 - 6 Aug 2025

Cited by 8 | Viewed by 1859

Abstract

The efficacy of botulinum toxin A (BoNT) in alleviating motor symptoms of cervical dystonia (CD) has been well established, and it is the treatment of choice in this disease. Lately, the effect of BoNT on non-motor symptoms (NMS) such as cognitive function, depression, [...] Read more.

The efficacy of botulinum toxin A (BoNT) in alleviating motor symptoms of cervical dystonia (CD) has been well established, and it is the treatment of choice in this disease. Lately, the effect of BoNT on non-motor symptoms (NMS) such as cognitive function, depression, anxiety, pain, and sleep disturbance has been observed in patients with CD. A comprehensive clinical and functional assessment of motor (dystonia severity, gait) and non-motor symptoms (cognitive functions, depression, anxiety, sleep, and pain) has been performed in a total of 34 adult patients with cervical dystonia before and after BoNT treatment. Results have also been compared to a control group. Significant improvements in the scales assessing dystonia severity have been observed, which is in line with previous studies on the effect of BoNT on motor symptoms in dystonia. Interestingly, the results also clearly indicate that BoNT has a positive effect on NMS. Among the studied non-motor domains, depression and cognitive functions improved the most after the treatment procedure. The study highlights the potential of BoNT to positively influence non-motor symptoms in patients with cervical dystonia, although its effect on various NMS is not equal. Full article

(This article belongs to the Special Issue Advances in the Treatment of Movement Disorders with Botulinum Toxins)

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23 pages, 9844 KB

Open AccessEditor’s ChoiceArticle

Mechanistic Exploration of Aristolochic Acid I-Induced Hepatocellular Carcinoma: Insights from Network Toxicology, Machine Learning, Molecular Docking, and Molecular Dynamics Simulation

by Tiantaixi Tu, Tongtong Zheng, Hangqi Lin, Peifeng Cheng, Ye Yang, Bolin Liu, Xinwang Ying and Qingfeng Xie

Toxins 2025, 17(8), 390; https://doi.org/10.3390/toxins17080390 - 5 Aug 2025

Cited by 6 | Viewed by 3163

Abstract

This study explores how aristolochic acid I (AAI) drives hepatocellular carcinoma (HCC). We first employ network toxicology and machine learning to map the key molecular target genes. Next, our research utilizes molecular docking to evaluate how AAI binds to these targets, and finally [...] Read more.

This study explores how aristolochic acid I (AAI) drives hepatocellular carcinoma (HCC). We first employ network toxicology and machine learning to map the key molecular target genes. Next, our research utilizes molecular docking to evaluate how AAI binds to these targets, and finally confirms the stability and dynamics of the resulting complexes through molecular dynamics simulations. We identified 193 overlapping target genes between AAI and HCC through databases such as PubChem, OMIM, and ChEMBL. Machine learning algorithms (SVM-RFE, random forest, and LASSO regression) were employed to screen 11 core genes. LASSO serves as a rapid dimension-reduction tool, SVM-RFE recursively eliminates the features with the smallest weights, and Random Forest achieves ensemble learning through decision trees. Protein–protein interaction networks were constructed using Cytoscape 3.9.1, and key genes were validated through GO and KEGG enrichment analyses, an immune infiltration analysis, a drug sensitivity analysis, and a survival analysis. Molecular-docking experiments showed that AAI binds to each of the core targets with a binding affinity stronger than −5 kcal mol⁻¹, and subsequent molecular dynamics simulations verified that these complexes remain stable over time. This study determined the potential molecular mechanisms underlying AAI-induced HCC and identified key genes (CYP1A2, ESR1, and AURKA) as potential therapeutic targets, providing valuable insights for developing targeted strategies to mitigate the health risks associated with AAI exposure. Full article

(This article belongs to the Section Plant Toxins)

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22 pages, 3176 KB

Open AccessEditor’s ChoiceArticle

Maresin 2, a Specialized Pro-Resolution Lipid Mediator, Reduces Pain and Inflammation Induced by Bothrops jararaca Venom in Mice

by Kassyo L. S. Dantas, Beatriz H. S. Bianchini, Matheus D. V. da Silva, Maiara Piva, Joice M. da Cunha, Janaina M. Zanoveli, Fernanda C. Cardoso, Fabiana T. M. C. Vicentini, Camila R. Ferraz, Patricia B. Clissa, Rubia Casagrande and Waldiceu A. Verri

Toxins 2025, 17(8), 367; https://doi.org/10.3390/toxins17080367 - 25 Jul 2025

Cited by 3 | Viewed by 2335

Abstract

The venom of Bothrops jararaca (BjV) induces intense and prolonged pain, which is not alleviated by antivenom, along with hemorrhage and inflammation. In this study, we investigated the effects of the specialized pro-resolving lipid mediator (SPM) maresin 2 (MaR2) in a murine model [...] Read more.

The venom of Bothrops jararaca (BjV) induces intense and prolonged pain, which is not alleviated by antivenom, along with hemorrhage and inflammation. In this study, we investigated the effects of the specialized pro-resolving lipid mediator (SPM) maresin 2 (MaR2) in a murine model of BjV-evoked pain and inflammation. Mice received a single intraperitoneal (i.p.) injection of MaR2 30 min before the intraplantar BjV injection. MaR2 treatment significantly attenuated mechanical (electronic aesthesiometer) and thermal (hot plate) hyperalgesia in a dose-dependent manner. Additionally, MaR2 restored the balance for the hind-paw static weight distribution. When BjV (0.01, 0.1, and 1 μg) stimulus was administered intraperitoneally, pre-treatment with MaR2 (0.3, 1, or 3 ng) ameliorated mechanical and thermal hyperalgesia in a dose-dependent manner. Moreover, MaR2 (3 ng) effectively reduced the levels of myeloperoxidase activity and cytokines (TNF-α, IL-1β, and IL-6) and superoxide anion (O₂^•−) production induced by intraplantar injection of BjV while enhancing total antioxidant levels (ABTS scavenging). For the peritonitis model induced by BjV, MaR2 pretreatment decreased leukocyte recruitment, hemorrhage, nitric oxide (NO), and O₂^•− generation and gp91phox and inducible nitric oxide synthase (iNOS) mRNA expression. In conclusion, this study presents the first evidence that MaR2 effectively mitigated BjV-induced pain, hemorrhage, and inflammation. Full article

(This article belongs to the Section Animal Venoms)

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19 pages, 1475 KB

Open AccessEditor’s ChoiceArticle

Chemical and Morphological Constitutive Defensive Traits of Cyanobacteria Have Different Effects on the Grazing of a Small Tropical Cladoceran

by Luciana Machado Rangel, Marcella Coelho Berjante Mesquita, Shara Rosa de Barros, Vinicius Neres-Lima, Michael Ribas Celano, Mauro Cesar Palmeira Vilar, Sandra Maria Feliciano de Oliveira e Azevedo and Marcelo Manzi Marinho

Toxins 2025, 17(7), 343; https://doi.org/10.3390/toxins17070343 - 5 Jul 2025

Cited by 4 | Viewed by 1911

Abstract

Antipredator defenses of bloom-forming cyanobacteria species maximize their fitness but can reduce carbon and energy transfer efficiency to higher trophic levels, making them a key regulator of plankton communities in eutrophic waters. We investigated the grazing responses of the tropical cladoceran Moina micrura [...] Read more.

Antipredator defenses of bloom-forming cyanobacteria species maximize their fitness but can reduce carbon and energy transfer efficiency to higher trophic levels, making them a key regulator of plankton communities in eutrophic waters. We investigated the grazing responses of the tropical cladoceran Moina micrura to different strains of the cyanobacteria Microcystis aeruginosa and Planktothrix isothrix, using a good food source (green algae Mono-raphidium capricornutum) as a control. Both Microcystis strains grow as unicellular and are microcystins producers; however, this cyanotoxin was not detected on the filamentous Planktothrix strains. M. micrura ingested all cyanobacteria at reduced rates compared to single diets with Monoraphidium. In mixed diets, food type had a significant effect on grazing responses, which differed interspecifically. Planktothrix was more grazed than Microcystis strains. Feeding selectivity on Monoraphidium was negatively affected by the increase of cyanobacteria in the diet. We observed varied responses across treatments, ranging from feeding inhibition to different degrees of tolerance toward cyanobacteria, particularly in non-microcystin-producing species. We also highlight the selectivity of small tropical cladocerans, a pattern that is not yet well documented. These findings emphasize that studies incorporating phyto- and zooplankton with a history of coexistence can provide more meaningful insights into natural ecosystem dynamics. Full article

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

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20 pages, 1159 KB

Open AccessEditor’s ChoiceArticle

Assessing Alternaria Species and Related Mycotoxin Contamination in Wheat in Algeria: A Food Safety Risk

by Meriem Barkahoum Daichi, Mario Masiello, Miriam Haidukowski, Annalisa De Girolamo, Antonio Moretti, Amor Bencheikh, Noureddine Rouag and Stefania Somma

Toxins 2025, 17(6), 309; https://doi.org/10.3390/toxins17060309 - 18 Jun 2025

Cited by 12 | Viewed by 3982

Abstract

Alternaria species are important fungal pathogens occurring worldwide in wheat, causing both productive and qualitative losses, and posing a toxicological risk to human health due to the production of their mycotoxins in kernels. This study aimed to investigate the occurrence of Alternaria species [...] Read more.

Alternaria species are important fungal pathogens occurring worldwide in wheat, causing both productive and qualitative losses, and posing a toxicological risk to human health due to the production of their mycotoxins in kernels. This study aimed to investigate the occurrence of Alternaria species and their mycotoxins in 48 wheat grain samples collected from the northeast to the southeast of Algeria. Seventy-two representative Alternaria strains were molecularly analyzed using a multi-locus sequence approach and evaluated for their capability to produce mycotoxins under in vitro conditions. Alternaria alternata, representing 42% of the strains, was the dominant species, followed to a lesser extent by species included in the Infectoriae section (26%). In addition, three species not previously reported in Algerian wheat, A. eureka, A. consortialis and A. tellustris, were identified, accounting for 5% of the total strains. Mycotoxin analyses showed high contamination of grains with alternariol monomethyl ether, alternariol and tenuazonic acid, occurring in 75, 69 and 35% of the samples, respectively. Moreover, 41 out of 48 samples showed the co-occurrence of multiple Alternaria mycotoxins. This study provides, for the first, time a clear picture of the occurrence and the distribution of Alternaria species on wheat in Algeria. Finally, the extensive monitoring activities carried out revealed the great biodiversity of Alternaria species able to colonize wheat grains. Moreover, findings on mycotoxin contamination raise concerns about the significant mycotoxigenic risk in Algerian wheat, emphasizing the need for strict monitoring and regulatory measures on Alternaria mycotoxins in food and feed. Full article

(This article belongs to the Section Mycotoxins)

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18 pages, 4121 KB

Open AccessEditor’s ChoiceArticle

Defence Against Desiccation and Predation in Lophyohylini Casque-Headed Tree Frogs

by César Alexandre, Pedro L. Mailho-Fontana, Bianca C. L. F. Távora, Marta M. Antoniazzi and Carlos Jared

Toxins 2025, 17(6), 303; https://doi.org/10.3390/toxins17060303 - 16 Jun 2025

Cited by 2 | Viewed by 5392

Abstract

Casque-headed tree frogs (Lophyohylini) can have a very large and distinctive head characterised by hyperossification of their cranial skin. This type of skull was primarily associated with phragmosis, a behaviour in which the frog enters holes backwards and seals them with its head [...] Read more.

Casque-headed tree frogs (Lophyohylini) can have a very large and distinctive head characterised by hyperossification of their cranial skin. This type of skull was primarily associated with phragmosis, a behaviour in which the frog enters holes backwards and seals them with its head to prevent water loss in challenging environments. Further investigations revealed that hyperossification also gives rise to bony spines interspersed with skin poison glands. These peculiar anatomical features of the head make it challenging for predators to prey on the frogs in phragmosis. When bitten on the head, the bite pressure causes the spines to cross the poison glands, allowing the injection of toxins into the predator’s mouth. We studied the head morphology of different Lophyohylini species along with some characteristics of their cutaneous poison, both in the field and in the laboratory. These frogs exemplify distinct chemical defence strategies, highlighting the differences between venom and poison. Notably, some species can cause self-poisoning in predators by injecting poison (in this case, venom) through their head spines, similar to the use of fangs by snakes. Full article

(This article belongs to the Collection Evolution of Venom Systems)

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14 pages, 263 KB

Open AccessEditor’s ChoiceArticle

Evaluating Bias in Self-Reported Symptoms During a Cyanobacterial Algal Bloom

by John S. Reif, Rebecca Koszalinski, Malcolm M. McFarland, Michael L. Parsons, Rachael Schinbeckler, Judyta Kociolek, Alex Rockenstyre and Adam M. Schaefer

Toxins 2025, 17(6), 287; https://doi.org/10.3390/toxins17060287 - 6 Jun 2025

Cited by 3 | Viewed by 2102

Abstract

Algal blooms produced by cyanobacteria liberate microcystins and other toxins that create a public health hazard. During the 2018 bloom of Microcystis aeruginosa in Florida, USA, residential and recreational exposures were associated with an increased risk of self-reporting respiratory, gastrointestinal, or ocular symptoms [...] Read more.

Algal blooms produced by cyanobacteria liberate microcystins and other toxins that create a public health hazard. During the 2018 bloom of Microcystis aeruginosa in Florida, USA, residential and recreational exposures were associated with an increased risk of self-reporting respiratory, gastrointestinal, or ocular symptoms for 125 participants. Subsequently, 207 persons were interviewed between 2019 and 2024 in the absence of large-scale algal blooms and were considered non-exposed. Analyses of cyanotoxins and brevetoxins in water and air showed only intermittent, background levels of toxins during the non-bloom period. The purpose of this report was to compare symptom reporting between active bloom and non-bloom periods. The assessment of information bias from self-reported symptoms is an important issue in epidemiologic studies of harmful algal blooms. During the non-bloom period, no statistically significant associations with residential, recreational, or occupational exposures were found for any symptom group. Estimated risks for respiratory, gastrointestinal, and ocular symptoms, headache, and skin rash were significantly higher for persons sampled during the bloom than the non-bloom period with odds ratios (ORs) of 2.3 to 8.3. ORs for specific respiratory symptoms were also significantly elevated. After adjustment for confounders and multiple exposures in multivariable analyses, the differences in symptom reporting between bloom and non-bloom periods remained statistically significant. In summary, the use of self-reported symptoms in this epidemiologic study of exposure to a cyanobacterial algal bloom did not appear to introduce substantial information bias. Full article

(This article belongs to the Special Issue Prospective Studies on Harmful Cyanobacteria and Cyanotoxins)

15 pages, 2495 KB

Open AccessEditor’s ChoiceArticle

Palytoxin Signal in LC-MS and UV: Preliminary Investigation on the Effect of Solvent and Temperature

by Chiara Melchiorre, Michela Varra, Valeria Tegola, Valentina Miele and Carmela Dell’Aversano

Toxins 2025, 17(6), 286; https://doi.org/10.3390/toxins17060286 - 6 Jun 2025

Cited by 3 | Viewed by 2263

Abstract

Palytoxins (PLTXs) and ovatoxins (OVTXs) are a group of highly potent marine toxins that pose significant health risks through seafood contamination and environmental exposure. OVTX-producing algae have been linked to dermatitis and respiratory distress in Mediterranean beachgoers, while serious public health concerns are [...] Read more.

Palytoxins (PLTXs) and ovatoxins (OVTXs) are a group of highly potent marine toxins that pose significant health risks through seafood contamination and environmental exposure. OVTX-producing algae have been linked to dermatitis and respiratory distress in Mediterranean beachgoers, while serious public health concerns are related to PLTX accumulation in seafood. In 2009, the European Food Safety Authority highlighted the need for analytical detection methods of the PLTX group of toxins and for the preparation of reference materials. This study investigates the stability of the palytoxin signal using liquid chromatography tandem mass spectrometry (LC-MRM-MS) and UV-Vis spectrophotometry under different experimental conditions: three concentrations (10, 1, and 0.5 µg/mL), three methanol–water mixtures (10%, 50%, and 90%), and two temperatures (6 °C and 25 °C). The results showed that the PLTX signal response is significantly influenced by the experimental conditions used. LC-MRM-MS analysis revealed the optimal response of PLTX in 50% and 90% MeOH at 25 °C, with minimal signal loss occurring over 16 h (9% and 6%). UV-Vis data indicated reduced absorbance in 10% MeOH, but a stable spectral intensity over 21 h in all the tested solvent mixtures. These results underscore the necessity of carefully controlled experimental conditions to ensure accurate and reproducible PLTX detection in environmental and food safety monitoring. Full article

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

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28 pages, 4731 KB

Open AccessEditor’s ChoiceArticle

Time-Resolved Visualization of Cyanotoxin Synthesis via Labeling by the Click Reaction in the Bloom-Forming Cyanobacteria Microcystis aeruginosa and Planktothrix agardhii

by Rainer Kurmayer and Rubén Morón Asensio

Toxins 2025, 17(6), 278; https://doi.org/10.3390/toxins17060278 - 3 Jun 2025

Cited by 2 | Viewed by 1863

Abstract

In non-ribosomal peptide synthesis of cyanobacteria, promiscuous adenylation domains allow the incorporation of clickable non-natural amino acids into peptide products—namely into microcystins (MCs) or into anabaenopeptins (APs): 4-azidophenylalanine (Phe-Az), N-propargyloxy-carbonyl-L-lysine (Prop-Lys), or O-propargyl-L-tyrosine (Prop-Tyr). Subsequently, chemo-selective labeling is used to visualize [...] Read more.

In non-ribosomal peptide synthesis of cyanobacteria, promiscuous adenylation domains allow the incorporation of clickable non-natural amino acids into peptide products—namely into microcystins (MCs) or into anabaenopeptins (APs): 4-azidophenylalanine (Phe-Az), N-propargyloxy-carbonyl-L-lysine (Prop-Lys), or O-propargyl-L-tyrosine (Prop-Tyr). Subsequently, chemo-selective labeling is used to visualize the clickable cyanopeptides using Alexa Fluor 488 (A488). In this study, the time-lapse build up or decline of azide- or alkyne-modified MCs or APs was visualized during maximum growth, specifically MC biosynthesis in Microcystis aeruginosa and AP biosynthesis in Planktothrix agardhii. Throughout the time-lapse build up or decline, the A488 signal occurred with heterogeneous intracellular distribution. There was a fast increase or decrease in the A488 signal for either Prop-Tyr or Prop-Lys, while a delayed or unobservable A488 signal for Phe-Az was related to increased cell size as well as a reduction in growth and autofluorescence. The proportion of clickable MC/AP in peptide extracts as recorded by a chemical–analytical technique correlated positively with A488 labeling intensity quantified via laser-scanning confocal microscopy for individual cells or via flow cytometry at the population level. It is concluded that chemical modification of MC/AP can be used to track intracellular dynamics in biosynthesis using both analytical chemistry and high-resolution imaging. Full article

(This article belongs to the Special Issue Harmful Algae in a Changing World: Where Did We Come from and Where Are We Going?)

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28 pages, 7518 KB

Open AccessEditor’s ChoiceArticle

Probing Bacterial Interactions with the Schistosoma mansoni-Killing Toxin Biomphalysin via Atomic Force Microscopy and Single Molecule Force Spectroscopy

by Jihen Zouaoui, Pierre Poteaux, Audrey Beaussart, Nicolas Lesniewska, David Duval and Jérôme F. L. Duval

Toxins 2025, 17(6), 269; https://doi.org/10.3390/toxins17060269 - 27 May 2025

Cited by 2 | Viewed by 2513

Abstract

Recent work has identified biomphalysin (BM) protein from the snail Biomphalaria glabrata as a cytolytic toxin against the Schistosoma mansoni parasite. Ex vivo interactome studies further evidenced BM’s ability to bind bacterial outer membrane proteins, but its specific antibacterial mechanisms and selectivity remain [...] Read more.

Recent work has identified biomphalysin (BM) protein from the snail Biomphalaria glabrata as a cytolytic toxin against the Schistosoma mansoni parasite. Ex vivo interactome studies further evidenced BM’s ability to bind bacterial outer membrane proteins, but its specific antibacterial mechanisms and selectivity remain unclear. Accordingly, this study aims to elucidate the interaction between BM and two model bacteria with distinct cell surface architectures: Escherichia coli (Gram−) and Micrococcus luteus (Gram+). Employing a multiscale approach, we used in vivo single-molecule force spectroscopy (SMFS) to probe molecular interactions at the single cell level. Combined with cell aggregation assays, immunoblotting and Atomic Force Microscopy (AFM) imaging, SMFS results evidenced a selective interaction of BM from snail plasma with M. luteus but not E. coli. Exposure of M. luteus to BM compromised cell surface integrity and induced cell aggregation. These effects correlated with a patch-like distribution of BM on M. luteus reminiscent of pore-forming toxins, as revealed by the anti-BM antibody-functionalized AFM tip. Overall, this work highlights the utility of SMFS in dissecting host–pathogen molecular dialogs. It reveals BM’s selective action against M. luteus, potentially via surface clustering, and it shows spatially heterogeneous responses to the toxin within and between individual cells. Full article

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

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16 pages, 4520 KB

Open AccessEditor’s ChoiceArticle

StingReady: A Novel Device for Controlled Insect Sting Challenge—From Field Capture to Clinical Application

by Xesús Feás, Margarita Armisén, Sara López-Freire, Manuela Alonso-Sampedro and Carmen Vidal

Toxins 2025, 17(6), 260; https://doi.org/10.3390/toxins17060260 - 22 May 2025

Cited by 2 | Viewed by 6293

Abstract

Reliable assessment of protection in venom immunotherapy (VIT) patients remains a clinical challenge, especially due to the limitations of conventional sting challenge tests (SCTs), which require complex insect handling and may compromise test accuracy. This study introduces StingReady, a novel, user-friendly device designed [...] Read more.

Reliable assessment of protection in venom immunotherapy (VIT) patients remains a clinical challenge, especially due to the limitations of conventional sting challenge tests (SCTs), which require complex insect handling and may compromise test accuracy. This study introduces StingReady, a novel, user-friendly device designed to streamline the SCT process by enabling safe, efficient, and minimally manipulative exposure to hymenopteran stings. For the first time, StingReady was applied to conduct SCTs with Vespa velutina, an invasive hornet species of increasing clinical relevance. The device was tested in a real-world setting at Belvís Park in Santiago de Compostela, Spain, where hornets were successfully captured and transported to the hospital without anesthesia or limb removal. The design features adjustable mesh sizes, allowing compatibility with various hymenopteran taxa. Using StingReady, nine patients underwent SCTs with no need for direct insect handling during the hospital procedure. The process improved patient safety and comfort while preserving the insect’s natural stinging behavior, thereby enhancing test reliability. This study demonstrates that StingReady significantly improves SCT methodology, offering a practical, reproducible, and ethically sound alternative for evaluating VIT efficacy across diverse hymenopteran species. Full article

(This article belongs to the Section Animal Venoms)

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15 pages, 3111 KB

Open AccessEditor’s ChoiceArticle

The Impact of Biocontrol Agents on the Metabolome of Penicillium nordicum Strains and Its Relation to Ochratoxin A Production on Dry-Cured Ham

by Eva Cebrián, Elia Roncero, João Luz, Mar Rodríguez, Marta Sousa Silva, Carlos Cordeiro and Félix Núñez

Toxins 2025, 17(5), 236; https://doi.org/10.3390/toxins17050236 - 9 May 2025

Viewed by 1844

Abstract

Throughout the process of dry-cured ham, moulds such as P. nordicum, a producer of ochratoxin A (OTA), grow on its surface. The use of combined biocontrol agents (BCAs) is a promising strategy for controlling this hazard. The goal of this study is [...] Read more.

Throughout the process of dry-cured ham, moulds such as P. nordicum, a producer of ochratoxin A (OTA), grow on its surface. The use of combined biocontrol agents (BCAs) is a promising strategy for controlling this hazard. The goal of this study is to assess the effect of D. hansenii, S. xylosus, and P. chrysogenum as BCAs on the metabolome of two strains of P. nordicum and to understand the differences between both strains. Each ochratoxigenic strain was inoculated both individually and in combination with the BCAs onto ham for 30 days under the environmental conditions experienced during traditional ripening. Untargeted metabolomics was performed through mass spectrometry using a Q-Exactive Plus Orbitrap. The BCAs caused alterations in the metabolomes of both ochratoxigenic moulds, mainly in phenylalanine catabolism and the valine, leucine, and isoleucine biosynthesis pathways, although with some differences. In the absence of the BCAs, the metabolomes of both types of P. nordicum were globally changed, despite these being moulds of the same species. In conclusion, these data help us to understand the differences between OTA-producing strains in dry-cured ham and confirm the need to demonstrate the efficacy of BCAs against a wide range of toxigenic moulds before they can be used to minimise OTA contamination in the meat industry. Full article

(This article belongs to the Special Issue Occurrence, Toxicity, Metabolism, Analysis and Control of Mycotoxins)

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11 pages, 4941 KB

Open AccessEditor’s ChoiceArticle

Consistent Killers: Conservation of Thrombin-Like Action on Fibrinogen by Bushmaster (Lachesis Species) Venoms Underpins Broad Antivenom Cross-Reactivities

by Lee Jones and Bryan G. Fry

Toxins 2025, 17(5), 224; https://doi.org/10.3390/toxins17050224 - 2 May 2025

Cited by 1 | Viewed by 3429

Abstract

Snakebite represents a significant public health challenge in Central and South America, with Lachesis (Bushmaster) species posing unique clinical challenges due to their severe envenomation effects arising from a combination of potent venom and copious venom yields. Using in vitro coagulation assays, we [...] Read more.

Snakebite represents a significant public health challenge in Central and South America, with Lachesis (Bushmaster) species posing unique clinical challenges due to their severe envenomation effects arising from a combination of potent venom and copious venom yields. Using in vitro coagulation assays, we analyzed the coagulotoxic venom effects from four distinct localities: L. muta from Surinam and French Guiana and L. stenophrys from Costa Rica and Panama. This study examined the venom’s impact on human plasma and fibrinogen and evaluated the efficacy of two regionally available antivenoms (PoliVal-ICP and Antivipmyn-Tri) in neutralizing the pathophysiological effects. Our results demonstrated a remarkable consistency in the pseudo-procoagulant venom activity (also known as: thrombin-like) across different species and localities. Antivenom efficacy testing revealed that both the PoliVal-ICP and Antivipmyn-Tri antivenoms effectively neutralized the venom effects across localities for both species, with the ICP antivenom showing the highest neutralization capacity. These toxicology findings highlight the biochemical conservation of venom composition across Lachesis species which underpins effective cross-neutralization in antivenom treatment. Full article

(This article belongs to the Section Animal Venoms)

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16 pages, 3855 KB

Open AccessEditor’s ChoiceArticle

A 20-Year Retrospective Analysis of Plant Poisoning Cases at the Naval Hospital, Varna, Bulgaria

by Stanila Stoeva-Grigorova, Maya Radeva-Ilieva, Stela Dragomanova, Gabriela Kehayova, Simeonka Dimitrova, Simeon Marinov, Petko Marinov, Marieta Yovcheva, Diana Ivanova and Snezha Zlateva

Toxins 2025, 17(4), 197; https://doi.org/10.3390/toxins17040197 - 12 Apr 2025

Cited by 2 | Viewed by 4406

Abstract

The nature and epidemiology of plant intoxications are still not well understood, with recent data being limited. The present study aims to report cases of plant poisoning in the clinical practice of the Clinical Toxicology Department at the Naval Hospital—Varna, Bulgaria, over a [...] Read more.

The nature and epidemiology of plant intoxications are still not well understood, with recent data being limited. The present study aims to report cases of plant poisoning in the clinical practice of the Clinical Toxicology Department at the Naval Hospital—Varna, Bulgaria, over a 20-year period (2003–2023). A documentary retrospective analysis of the hospitalized cases of poisoning with poisonous plants and their grouping into toxidromes was performed. During the study period, patients with plant poisoning admitted to our hospital unit accounted for 0.35% of a total of 12,857 hospitalized individuals. The distribution across the toxidromes based on clinical presentation revealed the highest frequency of anticholinergic, cyanogen, and ricin toxidromes. The majority of the intoxications resulted from unintentional exposure to plant toxins in adult individuals. Most cases followed a mild to severe clinical course, with patient discharge occurring between 2 and 5 days. No fatalities were recorded, thanks to the reported treatment methods. A relatively low incidence of plant-related poisonings was observed, with their predominant manifestations affecting the gastrointestinal, nervous, and cardiovascular systems. Increased reporting of epidemiological data and clinical experiences in the management of plant intoxications would substantially enhance researchers’ understanding of them and facilitate the development of a standardized treatment protocol. Full article

(This article belongs to the Section Plant Toxins)

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17 pages, 1736 KB

Open AccessEditor’s ChoiceArticle

Electrical Cell Impedance Sensing (ECIS): Feasibility of a Novel In Vitro Approach to Studying Venom Toxicity and Potential Therapeutics

by Abhinandan Choudhury, Kaitlin Linne, Tommaso C. Bulfone, Tanvir Hossain, Abu Ali Ibn Sina, Philip L. Bickler, Bryan G. Fry and Matthew R. Lewin

Toxins 2025, 17(4), 193; https://doi.org/10.3390/toxins17040193 - 11 Apr 2025

Cited by 3 | Viewed by 3222

Abstract

Snakebite envenoming is often discussed in terms of lethality and limb loss, but local tissue injury and coagulotoxic effects of venom are significantly more common acute manifestations of snakebite envenoming (SBE). Local tissue injury and the hemorrhagic and coagulotoxic effects of venom are [...] Read more.

Snakebite envenoming is often discussed in terms of lethality and limb loss, but local tissue injury and coagulotoxic effects of venom are significantly more common acute manifestations of snakebite envenoming (SBE). Local tissue injury and the hemorrhagic and coagulotoxic effects of venom are challenging to study in live animals and can be ethically fraught due to animal welfare concerns such that attention to the 3Rs of animal welfare motivates the development of in vitro techniques in this arena. Herein, we tested the use of a wound-healing study technique known as Electric Cell-Substrate Impedance Sensing (ECIS) to assess populations of cultured cells exposed to venom with or without sPLA₂ and/or metalloprotease inhibitors (varespladib and marimastat, respectively). For comparison, the StarMax coagulation analyzer for coagulotoxicity was further used to evaluate the venoms and the neutralizing capabilities of the abovementioned direct toxin inhibitors (DTIs) against the same venoms examined using ECIS. Three viper and three elapid venoms that were examined for their effects on H1975 cells were Agkistrodon contortrix (Eastern Copperhead), Crotalus helleri (Southern Pacific Rattlesnake), and Vipera ammodytes (Horned Viper) and Naja atra (Chinese Cobra), Naja mossambica (Mozambique Spitting Cobra), and Naja nigricollis (Black-necked Spitting Cobra), respectively. The combination of cellular and coagulation techniques appears to usefully discriminate the in vitro capabilities and limitations of specific inhibitors to inhibit specific venom effects. This study suggests that ECIS with or without concomitant coagulation testing is a feasible method to generate reproducible, meaningful preclinical data and could be used with any type of cell line. Importantly, this approach is both quantitative and has the potential of reducing animal use and suffering during the evaluation of potential therapeutics. To further evaluate the potential of this method, rescue studies should be performed. Full article

(This article belongs to the Special Issue Venoms and Drugs)

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23 pages, 6254 KB

Open AccessEditor’s ChoiceArticle

Computational Immunogenetic Analysis of Botulinum Toxin A Immunogenicity and HLA Gene Haplotypes: New Insights

by Eqram Rahman, Parinitha Rao, Munim Ahmed, William Richard Webb and Jean D. A. Carruthers

Toxins 2025, 17(4), 182; https://doi.org/10.3390/toxins17040182 - 6 Apr 2025

Cited by 19 | Viewed by 5184

Abstract

Botulinum toxin A (BoNT-A) is widely used in both therapeutic and aesthetic settings; however, the formation of neutralizing antibodies (NAbs) remains a critical concern, leading to treatment failure. Immunogenic responses are known to vary between individuals due to HLA polymorphisms. Although some claim [...] Read more.

Botulinum toxin A (BoNT-A) is widely used in both therapeutic and aesthetic settings; however, the formation of neutralizing antibodies (NAbs) remains a critical concern, leading to treatment failure. Immunogenic responses are known to vary between individuals due to HLA polymorphisms. Although some claim that neurotoxin-associated proteins (NAPs) shield BoNT-A from immune detection or are themselves immunogenic, there is limited molecular evidence supporting either view. This study applies computational immunogenetics to explore BoNT-A immunogenicity, focusing on HLA binding and the influence of accessory proteins. Epitope mapping, molecular docking, and HLA binding predictions were used to evaluate interactions between BoNT-A epitopes and selected class II HLA alleles (HLA-DQA1*01:02, HLA-DQA1*03:03, HLA-DQB1*06:04, HLA-DQB1*03:01, and HLA-DRB1*15:01). To assess the potential immunomodulatory role of NAPs, molecular dynamics (MD) simulations, solvent-accessible surface area (SASA) analysis, and electrostatic potential mapping were also conducted. Key epitopes—L11, N25, and C10—showed strong binding affinities to HLA-DQA1*01:02, HLA-DQB1*06:04, and HLA-DQA1*03:03, indicating a potential immunodominant role. NAPs did not obstruct these epitopes but slightly increased their exposure and appeared to stabilize the toxin structure. Electrostatic mapping and binding free energy calculations suggested no significant immunogenic shift in the presence of NAPs. BoNT-A immunogenicity appears to be influenced by HLA allele variability, reinforcing the value of patient-specific genetic profiling. The presumed immunogenic role of NAPs remains unsubstantiated at the molecular level, underscoring the need for evidence-based evaluation over commercial rhetoric. While these findings provide valuable molecular insight, it is important to acknowledge that they are derived entirely from in silico analyses. As such, experimental validation remains essential to confirm the immunological relevance of these predicted interactions. Nonetheless, this computational framework offers a rational basis for guiding future clinical research and the development of HLA-informed BoNT-A therapies. Full article

(This article belongs to the Section Bacterial Toxins)

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20 pages, 4179 KB

Open AccessEditor’s ChoiceArticle

Microbiological and Mycotoxicological Quality of Common Wheat in Romania in the Extremely Dry 2023–2024 Agricultural Year

by Valeria Gagiu, Elena Mirela Cucu, Alina Alexandra Dobre, Gina Pusa Pirvu, Oana Alexandra Oprea, Cristian Mihai Pomohaci, Elena Mateescu, Nastasia Belc and Doru Ioan Marin

Toxins 2025, 17(4), 154; https://doi.org/10.3390/toxins17040154 - 22 Mar 2025

Cited by 3 | Viewed by 6609

Abstract

This study examines the microbiological and mycotoxicological quality of common wheat in Romania in the extremely dry 2023–2024 agricultural year. Common wheat grown in the West Plain, Southern Hilly Area, Transylvania, and northern Moldavia (45–48° N, 21–27° E) had higher moisture content, water [...] Read more.

This study examines the microbiological and mycotoxicological quality of common wheat in Romania in the extremely dry 2023–2024 agricultural year. Common wheat grown in the West Plain, Southern Hilly Area, Transylvania, and northern Moldavia (45–48° N, 21–27° E) had higher moisture content, water activity, Fusarium-damaged kernels, and deoxynivalenol levels. This was due to moderate temperatures, abundant precipitation, and soil water reserves in May, followed by moderate drought from June to August. Conversely, common wheat from the Oltenia Plain, the Southern Plain, and southern Moldavia (43–46° N, 23–28° E) had the lowest contamination levels, attributed to extreme temperatures and drought during June–August. Common wheat from Dobrogea (45° N, 28° E) showed the highest total fungi contamination, which was influenced by precipitation at harvest. Although microbiological and mycotoxicological contamination was low, it negatively affected the physico-chemical and sensory–colorimetric parameters of common wheat, particularly in the West Plain, Oltenia Plain, and Dobrogea. Consequently, there could be significant economic losses for farmers, storekeepers, millers, and bakers, as well as a decline in the quality of finished foods. Moreover, the coexistence of deoxynivalenol and total aflatoxins in common wheat grown in the northwest of the country indicates the spread of contamination due to dry conditions and climate change. Full article

(This article belongs to the Collection Impact of Climate Change on Fungal Population and Mycotoxins)

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14 pages, 888 KB

Open AccessEditor’s ChoiceArticle

Snake Venom Makeover: Age-Dependent Variations in Procoagulant Biochemistry of Egyptian Saw-Scaled Viper (Echis pyramidum pyramidum) Venom

by Alex Barker, Lee Jones, Lachlan A. Bourke, Lorenzo Seneci, Abhinandan Chowdhury, Aude Violette, Rudy Fourmy, Raul Soria, Matt Aldridge and Bryan G. Fry

Toxins 2025, 17(3), 149; https://doi.org/10.3390/toxins17030149 - 19 Mar 2025

Cited by 8 | Viewed by 4790

Abstract

Echis species (saw-scaled vipers) are WHO Category 1 medically significant venomous snakes with potent procoagulant venoms, which cause lethal venom-induced consumptive coagulopathy in human victims. Despite clinical presentations of bites varying significantly between individuals within the same species, the contribution of age-related changes [...] Read more.

Echis species (saw-scaled vipers) are WHO Category 1 medically significant venomous snakes with potent procoagulant venoms, which cause lethal venom-induced consumptive coagulopathy in human victims. Despite clinical presentations of bites varying significantly between individuals within the same species, the contribution of age-related changes in the venom biochemistry has not been investigated. This study investigated the ontogenetic changes in Echis pyramidum pyramidum venom and its impact on therapeutic efficacy. The efficacy of various antivenoms (Echitab, Echitab+ ICP, Inosan MENA, Inosan Pan African, and SAVP-Echis) was tested against both venom phenotypes. While both neonate and adult venoms were procoagulant, there were differences in the underlying biochemistry. Neonate venom was found to potently pathophysiologically activate Factor VII and Factor X, and to a lesser degree Factor XII. In contrast, adult venom was a slower clotter, less potent in activating FVII, equipotent with neonate venom on FXII, and inactive on FX. This is the first documentation of FVII and FXII activation for any Echis venom. The significant ontogenetic toxicological variations in Echis species were shown to impact antivenom efficacy. Among the tested antivenoms, SAVP-Echis was the most effective against both venom phenotypes, with adult venom being better neutralized. These findings suggest the need for a reconsideration of venom mixture selection in antivenom production through the inclusion of neonate venom. Additionally, the results indicate differential ontogenetic predatory ecology, providing a foundation for future natural history investigations. Full article

(This article belongs to the Special Issue Snake Bite and Related Injury)

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22 pages, 629 KB

Open AccessEditor’s ChoiceArticle

Investigation into Paralytic Shellfish Toxins and Microcystins in Seabirds from Portugal

by Lucía Soliño, Andrew D. Turner, Begoña Ben-Gigirey, Ryan P. Alexander, Karl J. Dean, Robert G. Hatfield, Benjamin H. Maskrey and María V. Mena Casero

Toxins 2025, 17(3), 135; https://doi.org/10.3390/toxins17030135 - 13 Mar 2025

Cited by 1 | Viewed by 2561

Abstract

Microalgae form the basis of marine food webs, essential in sustaining top predators including seabirds. However, certain species of microalgae synthesize biotoxins, which can accumulate in shellfish and fish and may cause harm to marine animals feeding on them. Toxins produced by dinoflagellates [...] Read more.

Microalgae form the basis of marine food webs, essential in sustaining top predators including seabirds. However, certain species of microalgae synthesize biotoxins, which can accumulate in shellfish and fish and may cause harm to marine animals feeding on them. Toxins produced by dinoflagellates have been previously observed to be poisonous to seabirds. Also, in freshwater and brackish habitats, cyanobacteria have caused bird mortality events. In this work, we analyze the prevalence of six families of biotoxins (paralytic shellfish toxins (PSTs), microcystins (MCs), anatoxins, amnesic shellfish toxins (ASTs), cylindrospermopsin, and tetrodotoxins (TTXs)) in 340 samples from 193 wild birds admitted to a wildlife rehabilitation centre in south Portugal. Furthermore, we consider the clinical picture and signs of 17 birds that presented quantifiable levels of biotoxins in their tissues. The relationship between toxin burdens and the symptomatology observed, as well as possible biotoxin sources, are discussed. Based on previously published research data, we conclude that, in these birds, the biotoxins are unlikely to be the only cause of death but might contribute to some extent to a reduction in birds’ fitness. Full article

(This article belongs to the Special Issue Harmful Algal Blooms in Waters: Characterization, Monitoring and Management)

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15 pages, 968 KB

Open AccessEditor’s ChoiceArticle

Learnings from Separate Aconitum Poisonings in British Columbia and Ontario, Canada in 2022

by Lorraine McIntyre, Stefanie Georgopoulos, Dorianna Simone, Emily Newhouse, JoAnne Fernandes, David A. McVea, Arnold Fok, Ania-Maria McIntyre, Bryn Shurmer, Marie-Claude Gagnon, Michael Chan, Marina Chiaravalloti, Nikita Saha Turna, Debra Kent, Dennis Leong, Katherine Paphitis, Christina Lee and the Outbreak Investigation Teams

Toxins 2025, 17(3), 125; https://doi.org/10.3390/toxins17030125 - 7 Mar 2025

Cited by 4 | Viewed by 9917

Abstract

Background: Three aconitine poisoning events occurred in two Canadian provinces in 2022: one in British Columbia (BC) and two in Ontario (ON). Aconitine is a potent alkaloid found in several species of the plant Aconitum, containing cardiotoxins and neurotoxins. It is used [...] Read more.

Background: Three aconitine poisoning events occurred in two Canadian provinces in 2022: one in British Columbia (BC) and two in Ontario (ON). Aconitine is a potent alkaloid found in several species of the plant Aconitum, containing cardiotoxins and neurotoxins. It is used in traditional Chinese medicine (TCM) for pain management, and in powdered form, Aconitum is similar in appearance to sand ginger (Kaempferia galanga), which can lead to poisonings from misidentification and mislabeling. Methods: Aconitine poisoning is rare in Canada; here, we compare communications, collaborations, laboratory testing options and actions during investigations. Results: Fourteen cases occurred from the consumption of sand ginger: in BC (n = 2), purchased at an Asian health food store; in ON (n = 11), Kaempferia galanga powder (KGP) spices were used to prepare meals at a restaurant, and in one ON case, KGP was purchased. Traceback found product imported from China contained aconitine levels ranging from 1304 to 5500 ppm. Later investigations revealed mislabeling of Aconitum as KGP from the same imported lot (January 2020). Plant DNA testing found no KGP in any spice packets, including lots not linked to illness, suggestive of adulteration. Conclusion: Method development for aconitine in BC led to an improved response time for testing in ON. BC and ON updated outbreak response protocols and communications. Full article

(This article belongs to the Special Issue Plant Toxin Emergency)

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18 pages, 3357 KB

Open AccessEditor’s ChoiceArticle

by Pennapa Thongararm, Marisa Chancharoen, Nutchapong Suwanwong, Somsak Ruchirawat, Mathuros Ruchirawat, Bogdan I. Fedeles, Robert G. Croy and John M. Essigmann

Toxins 2025, 17(3), 112; https://doi.org/10.3390/toxins17030112 - 27 Feb 2025

Cited by 3 | Viewed by 3393

Abstract

Aflatoxin B₁ (AFB₁) and sterigmatocystin (ST) are mycotoxins that pose significant threats to human and animal health owing to their mutagenic, carcinogenic, and toxic properties. They are structurally similar and widely believed to exert their biological effects via the generation [...] Read more.

Aflatoxin B₁ (AFB₁) and sterigmatocystin (ST) are mycotoxins that pose significant threats to human and animal health owing to their mutagenic, carcinogenic, and toxic properties. They are structurally similar and widely believed to exert their biological effects via the generation of DNA-damaging epoxides at their respective terminal furan rings. Despite structural identity in the warhead portion of each toxin, this work shows that distal parts of each molecule are responsible for the distinctive mutational fingerprints seen in gptΔ C57BL/6J mouse embryo fibroblasts (MEFs). The two toxins differ structurally in the puckered cyclopentenone ring of AFB₁ and in the planar xanthone functionality of ST. While both toxins mainly induce GC→TA mutations, the aforementioned differences in structure apparently trigger unique patterns of mutations, as revealed by high-resolution duplex sequencing of MEF genomes. AFB₁ is more mutagenic than ST and displays its transversion mutations in a pattern with primary and secondary hotspots (underscored) in 5′-CGC-3′ and 5′-CGG-3′ contexts, respectively. ST displays a modest 5′-CGG-3′ hotspot while its other GC→TA transversions are more uniformly distributed in a pattern resembling established oxidative stress mutational spectra. This research delineates the mutational spectra of AFB₁ and ST, establishing these patterns as possible early-onset biomarkers of exposure. Full article

(This article belongs to the Special Issue 65 Years On from Aflatoxin Discovery—a Themed Issue in Honor of Professor John D. Groopman)

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17 pages, 5927 KB

Open AccessEditor’s ChoiceArticle

Pulsed Electric Field Induces Significant Changes in the Metabolome of Fusarium Species and Decreases Their Viability and Toxigenicity

by Adam Behner, Jana Palicova, Anna-Hirt Tobolkova, Nela Prusova and Milena Stranska

Toxins 2025, 17(1), 33; https://doi.org/10.3390/toxins17010033 - 11 Jan 2025

Cited by 4 | Viewed by 3408

Abstract

Fusarium fungi are widespread pathogens of food crops, primarily associated with the formation of mycotoxins. Therefore, effective mitigation strategies for these toxicogenic microorganisms are required. In this study, the potential of pulsed electric field (PEF) as an advanced technology of increasing use in [...] Read more.

Fusarium fungi are widespread pathogens of food crops, primarily associated with the formation of mycotoxins. Therefore, effective mitigation strategies for these toxicogenic microorganisms are required. In this study, the potential of pulsed electric field (PEF) as an advanced technology of increasing use in the food processing industry was investigated to minimize the viability of Fusarium pathogens and to characterize the PEF-induced changes at the metabolomic level. Spores of four Fusarium species (Fusarium culmorum, Fusarium graminearum, Fusarium poae, and Fusarium sporotrichioides) were treated with PEF and cultured on potato dextrose agar (PDA) plates. The viability of the Fusarium species was assessed by counting the colony-forming units, and changes in the mycotoxin content and metabolomic fingerprints were evaluated by using UHPLC-HRMS/MS instrumental analysis. For metabolomic data processing and compound identification, the MS-DIAL (v. 4.80)–MS-CleanR–MS-Finder (v. 3.52) software platform was used. As we found out, both fungal viability and the ability to produce mycotoxins significantly decreased after the PEF treatment for all of the species tested. The metabolomes of the treated and untreated fungi showed statistically significant differences, and PEF-associated biomarkers from the classes oxidized fatty acid derivatives, cyclic hexapeptides, macrolides, pyranocoumarins, carbazoles, and guanidines were identified. Full article

(This article belongs to the Section Mycotoxins)

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15 pages, 23082 KB

Open AccessEditor’s ChoiceArticle

Reduction in Renal Heme Oxygenase-1 Is Associated with an Aggravation of Kidney Injury in Shiga Toxin-Induced Murine Hemolytic-Uremic Syndrome

by Antonio N. Mestekemper, Wiebke Pirschel, Nadine Krieg, Maria K. Paulmann, Christoph Daniel, Kerstin Amann and Sina M. Coldewey

Toxins 2024, 16(12), 543; https://doi.org/10.3390/toxins16120543 - 14 Dec 2024

Cited by 1 | Viewed by 2505

Abstract

Hemolytic-uremic syndrome (HUS) is a systemic complication of an infection with Shiga toxin (Stx)-producing enterohemorrhagic Escherichia coli, primarily leading to acute kidney injury (AKI) and microangiopathic hemolytic anemia. Although free heme has been found to aggravate renal damage in hemolytic diseases, the [...] Read more.

Hemolytic-uremic syndrome (HUS) is a systemic complication of an infection with Shiga toxin (Stx)-producing enterohemorrhagic Escherichia coli, primarily leading to acute kidney injury (AKI) and microangiopathic hemolytic anemia. Although free heme has been found to aggravate renal damage in hemolytic diseases, the relevance of the heme-degrading enzyme heme oxygenase-1 (HO-1, encoded by Hmox1) in HUS has not yet been investigated. We hypothesized that HO-1, also important in acute phase responses in damage and inflammation, contributes to renal pathogenesis in HUS. The effect of tamoxifen-induced Hmox1 gene deletion on renal HO-1 expression, disease progression and AKI was investigated in mice 7 days after HUS induction. Renal HO-1 levels were increased in Stx-challenged mice with tamoxifen-induced Hmox1 gene deletion (Hmox1^R26Δ/Δ) and control mice (Hmox1^lox/lox). This HO-1 induction was significantly lower (−43%) in Hmox1^R26Δ/Δ mice compared to Hmox1^lox/lox mice with HUS. Notably, the reduced renal HO-1 expression was associated with an exacerbation of kidney injury in mice with HUS as indicated by a 1.7-fold increase (p = 0.02) in plasma neutrophil gelatinase-associated lipocalin (NGAL) and a 1.3-fold increase (p = 0.06) in plasma urea, while other surrogate parameters for AKI (e.g., periodic acid Schiff staining, kidney injury molecule-1, fibrin deposition) and general disease progression (HUS score, weight loss) remained unchanged. These results indicate a potentially protective role of HO-1 in the pathogenesis of Stx-mediated AKI in HUS. Full article

(This article belongs to the Section Bacterial Toxins)

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19 pages, 5894 KB

Open AccessEditor’s ChoiceArticle

Application of Pulsed Electric Field During Malting: Impact on Fusarium Species Growth and Mycotoxin Production

by Nela Prusova, Marcel Karabin, Lukas Jelinek, Jana Chrpova, Jaroslava Ovesna, Pavel Svoboda, Tereza Dolezalova, Adam Behner, Jana Hajslova and Milena Stranska

Toxins 2024, 16(12), 537; https://doi.org/10.3390/toxins16120537 - 12 Dec 2024

Cited by 6 | Viewed by 2339

Abstract

The increasing contamination of cereals by micromycetes and mycotoxins during malting still poses an unresolved food safety problem. This study characterises the potential of the novel, rapidly developing food production technology of Pulsed Electric Field (PEF) to reduce the viability of Fusarium fungi [...] Read more.

The increasing contamination of cereals by micromycetes and mycotoxins during malting still poses an unresolved food safety problem. This study characterises the potential of the novel, rapidly developing food production technology of Pulsed Electric Field (PEF) to reduce the viability of Fusarium fungi and the production of mycotoxins during malting. Barley, artificially inoculated with four Fusarium species, was treated by PEF with two different intensities and then malted using a standard Pilsner-type technology. Concentrations of fungi were quantified by RT-PCR, expression of fungal growth-related genes was assessed using mRNA sequencing, and mycotoxin levels were analysed by U-HPLC-HRMS/MS. Despite the different trends for micromycetes and mycotoxins after application of variously intense PEF conditions, significant reductions were generally observed. The greatest decrease was for F. sporotrichioides and F. poae, where up to six fold lower levels were achieved for malts produced from the PEF-treated barley when compared to the control. For F. culmorum and F. graminearum, up to a two-fold reduction in the PEF-generated malts was observed. These reductions mostly correlated with a decrease in relevant mycotoxins, specifically type A trichothecenes. Full article

(This article belongs to the Section Mycotoxins)

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13 pages, 6199 KB

Open AccessEditor’s ChoiceArticle

Bicistronic Vector Expression of Recombinant Jararhagin-C and Its Effects on Endothelial Cells

by Karla Fernanda Ferraz, Lhiri Hanna De Lucca Caetano, Daniele Pereira Orefice, Paula Andreia Lucas Calabria, Maisa Splendore Della-Casa, Luciana Aparecida Freitas-de-Sousa, Emidio Beraldo-Neto, Sabri Saeed Sanabani, Geraldo Santana Magalhães and Patricia Bianca Clissa

Toxins 2024, 16(12), 524; https://doi.org/10.3390/toxins16120524 - 3 Dec 2024

Cited by 3 | Viewed by 1852

Abstract

Jararhagin-C (JarC) is a protein from the venom of Bothrops jararaca consisting of disintegrin-like and cysteine-rich domains. JarC shows a modulating effect on angiogenesis and remodeling of extracellular matrix constituents, improving wound healing in a mouse experimental model. JarC is purified from crude [...] Read more.

Jararhagin-C (JarC) is a protein from the venom of Bothrops jararaca consisting of disintegrin-like and cysteine-rich domains. JarC shows a modulating effect on angiogenesis and remodeling of extracellular matrix constituents, improving wound healing in a mouse experimental model. JarC is purified from crude venom, and the yield is less than 1%. The aim of this work was to obtain the recombinant form of JarC and to test its biological activity. For this purpose, the bicistronic vector pSUMOUlp1 was used. This vector allowed the expression of the recombinant toxin JarC (rJarC) in fusion with the small ubiquitin-related modifier (SUMO) as well as the SUMO protease Ulp1. After expression, this protease was able to efficiently remove SUMO from rJarC inside the bacteria. rJarC free from SUMO was purified at the expected molecular mass and recognized by polyclonal anti-jararhagin antibodies. In terms of biological activity, both the native and recombinant forms showed no toxicity to the HUVEC cell line CRL1730 and were effective in modulating cell migration activity in the experimental in vitro model. These results demonstrate the successful production of rJarC and the preservation of its biological activity, which may facilitate further investigations into the therapeutic potential of this snake venom-derived protein. Full article

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

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14 pages, 920 KB

Open AccessEditor’s ChoiceArticle

Age Is Just a Number: Ontogenetic Conservation in Activation of Blood Clotting Factors VII, X, and XII by Caucasus Blunt-Nosed Viper (Macrovipera lebetina obtusa) Venoms

by Katrina Kempson, Abhinandan Chowdhury, Aude Violette, Rudy Fourmy, Raul Soria and Bryan G. Fry

Toxins 2024, 16(12), 520; https://doi.org/10.3390/toxins16120520 - 2 Dec 2024

Cited by 8 | Viewed by 5232

Abstract

This study examined the pathophysiological effects of venoms from neonate and adult specimens of the viperid snake Macrovipera lebetina obtusa, focusing on their ability to activate various blood clotting factors in human plasma. All venoms exhibited strong procoagulant properties. In concentration–response tests, [...] Read more.

This study examined the pathophysiological effects of venoms from neonate and adult specimens of the viperid snake Macrovipera lebetina obtusa, focusing on their ability to activate various blood clotting factors in human plasma. All venoms exhibited strong procoagulant properties. In concentration–response tests, the clotting potency of the neonate venoms fell within the range of their parents’ maximum clotting velocities and areas under the curve. Intriguingly, females were more potent than males within each age group, but this requires a larger sample size to confirm. Antivenom neutralization efficacy was equipotent across age groups. The venoms potently activated Factor X (FX) robustly, consistent with previous knowledge of this genus. For the first time, the ability to activate Factors VII (FVII) and XII (FXII) was identified in this genus, with FXII exhibiting particularly strong activation. The study found no significant ontogenetic variation in procoagulant venom potency on human plasma, convergent with the Daboia genus, the other large-bodied lineage within the Palearctic viperid clade. However, the activation of FXII and FVII reveals previously undocumented pathways in the procoagulant activity of these venoms, contributing to the broader understanding of venom evolution and its clinical impacts. These findings have implications for venom biodiscovery and the development of antivenoms, highlighting the complexity of clotting factor activation beyond traditional investigations that have myopically focused upon FX and prothrombin pathways, thereby underscoring the importance of exploring additional clotting factors. Full article

(This article belongs to the Section Animal Venoms)

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22 pages, 1624 KB

Open AccessEditor’s ChoiceArticle

Mycotoxin Challenge in Dairy Cows: Assessment of the Efficacy of an Anti-Mycotoxin Agent by Adopting an In Vitro Rumen Simulation Method

by Erica Fiorbelli, Marco Lapris, Michela Errico, Antonella Della Badia, Insaf Riahi, Gabriele Rocchetti and Antonio Gallo

Toxins 2024, 16(11), 490; https://doi.org/10.3390/toxins16110490 - 13 Nov 2024

Cited by 7 | Viewed by 4293

Abstract

To protect ruminants from the harmful effects of mycotoxins, anti-mycotoxin agents can be added to the dietary ration, thus guaranteeing animal health and production. Therefore, the objective of this study was to evaluate the in vitro ruminal initial sequestration (weak binding) and subsequent [...] Read more.

To protect ruminants from the harmful effects of mycotoxins, anti-mycotoxin agents can be added to the dietary ration, thus guaranteeing animal health and production. Therefore, the objective of this study was to evaluate the in vitro ruminal initial sequestration (weak binding) and subsequent desorption (strong binding) of an anti-mycotoxin agent based on a mixture of adsorbing material, turmeric and milk thistle extracts and yeast-based components to adsorb or bio-convert aflatoxins (AF), fumonisins B1 and B2 (FB), trichothecene deoxynivalenol (DON), T-2 and HT-2 toxins, and zearalenone (ZEN). Two doses were tested: Dose 1 simulated 30 mg/cow/d, while Dose 2 simulated 90 mg/cow/d of the anti-mycotoxin agent. Each treatment involved three analytical replicates at each of three incubation times (1, 4, and 24 h post-incubation), with two independent experimental runs providing experimental replicates. Analytical methods, including UHPLC-HRMS and multivariate analyses, were used to both quantify mycotoxin concentrations and reveal dose-dependent reductions, with statistical validations indicating significant changes in mycotoxin levels across both dose and time. The results indicated that the anti-mycotoxin agent was able to highly bind AFB1, T2, and HT-2 toxins since its concentration was always under the limit of detection (<1 ppb). Regarding ZEN (weak binding mean: 94.6%; strong binding mean: 62.4%) and FBs (weak binding mean: 58.7%; strong binding mean: 32.3%), orthogonal contrasts indicated that the anti-mycotoxin agent was able to effectively bind these toxins using Dose 1 (p < 0.05). This finding suggests that Dose 1 may be sufficient to achieve the targeted effect and that a further increase does not significantly improve the outcome. Regarding DON, a strong linear relationship was observed between dose and adsorption. However, the complex interactions between the mycotoxin, the ruminal environment, and the anti-mycotoxin agent made it difficult to establish a clear dose–effect relationship (p > 0.10). UHPLC-HRMS analysis identified over 1500 mass features in rumen samples, which were further analyzed to assess the effects of the anti-mycotoxin agent. Hierarchical clustering analysis (HCA) revealed significant changes in the untargeted metabolomic profiles of samples treated with mycotoxins compared to control samples, particularly after 24 h with the anti-mycotoxin treatments. Clear differences were noted between strong binding and weak binding samples. Further analysis using orthogonal partial least squares discriminant analysis (OPLS-DA) highlighted distinct metabolomic profiles, with stronger predictive ability in the strong binding group (Q² cumulative value of 0.57) compared to the weak binding group (0.30). The analysis identified 44 discriminant compounds in the strong binding model and 16 in the weak binding model. Seven compounds were common to both groups, while silibinin, known for its antioxidant and anti-inflammatory properties, was found among the unique compounds in the weak binding group. Overall, the findings suggest that both doses of the anti-mycotoxin agent significantly influenced the chemical profiles in the rumen, particularly enhancing the binding of mycotoxins, thereby supporting the role of phytogenic extracts in mitigating mycotoxin effects. Full article

(This article belongs to the Special Issue Mitigation and Detoxification Strategies of Mycotoxins)

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28 pages, 13740 KB

Open AccessEditor’s ChoiceArticle

A Novel P-III Metalloproteinase from Bothrops barnetti Venom Degrades Extracellular Matrix Proteins, Inhibits Platelet Aggregation, and Disrupts Endothelial Cell Adhesion via α5β1 Integrin Receptors to Arginine–Glycine–Aspartic Acid (RGD)-Containing Molecules

by Pedro Henrique de Caires Schluga, Debora Larangote, Ana Maria de Melo, Guilherme Kamienski Lobermayer, Daniel Torrejón, Luciana Souza de Oliveira, Valeria Gonçalves Alvarenga, Dan Erick Vivas-Ruiz, Silvio Sanches Veiga, Eladio Flores Sanchez and Luiza Helena Gremski

Toxins 2024, 16(11), 486; https://doi.org/10.3390/toxins16110486 - 9 Nov 2024

Cited by 5 | Viewed by 3658

Abstract

Viperid snake venoms are notably abundant in metalloproteinases (proteins) (SVMPs), which are primarily responsible for inducing hemorrhage and disrupting the hemostatic process and tissue integrity in envenomed victims. In this study, barnettlysin-III (Bar-III), a hemorrhagic P-III SVMP, was purified from the venom of [...] Read more.

Viperid snake venoms are notably abundant in metalloproteinases (proteins) (SVMPs), which are primarily responsible for inducing hemorrhage and disrupting the hemostatic process and tissue integrity in envenomed victims. In this study, barnettlysin-III (Bar-III), a hemorrhagic P-III SVMP, was purified from the venom of the Peruvian snake Bothrops barnetti. Bar-III has a molecular mass of approximately 50 kDa and is a glycosylation-dependent functional metalloproteinase. Some biochemical properties of Bar-III, including the full amino acid sequence deduced from its cDNA, are reported. Its enzymatic activity is increased by Ca²⁺ ions and inhibited by an excess of Zn²⁺. Synthetic metalloproteinase inhibitors and EDTA also inhibit its proteolytic action. Bar-III degrades several plasma and ECM proteins, including fibrin(ogen), fibronectin, laminin, and nidogen. Platelets play a key role in hemostasis and thrombosis and in other biological process, such as inflammation and immunity, and platelet activation is driven by the platelet signaling receptors, glycoprotein (GP)Ib-IX-V, which binds vWF, and GPVI, which binds collagen. Moreover, Bar-III inhibits vWF- and convulxin-induced platelet aggregation in human washed platelets by cleaving the recombinant A1 domain of vWF and GPVI into a soluble ectodomain fraction of ~55 kDa (sGPVI). Bar-III does not reduce the viability of cultured endothelial cells; however, it interferes with the adhesion of these cells to fibronectin, vitronectin, and RGD peptides, as well as their migration profile. Bar-III binds specifically to the surface of these cells, and part of this interaction involves α5β1 integrin receptors. These results contribute to a better comprehension of the pathophysiology of snakebite accidents/incidents and could be used as a tool to explore novel and safer anti-venom therapeutics. Full article

(This article belongs to the Section Animal Venoms)

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17 pages, 2038 KB

Open AccessEditor’s ChoiceArticle

Integrated Approach to Cyclopiazonic Acid Cytotoxicity Using In Vitro (2D and 3D Models) and In Silico Methods

by Carmen Martínez-Alonso, Luana Izzo, Yelko Rodríguez-Carrasco and María-José Ruiz

Toxins 2024, 16(11), 473; https://doi.org/10.3390/toxins16110473 - 3 Nov 2024

Cited by 7 | Viewed by 2738

Abstract

Cyclopiazonic acid (CPA) is an indole-tetramic acid neurotoxin produced by Aspergillus and Penicillium genera present mainly in fruit, cereals and nuts. This study compares the cytotoxicity produced by CPA after 24, 48 and 72 h of exposure using both monolayers and 3D spheroids [...] Read more.

Cyclopiazonic acid (CPA) is an indole-tetramic acid neurotoxin produced by Aspergillus and Penicillium genera present mainly in fruit, cereals and nuts. This study compares the cytotoxicity produced by CPA after 24, 48 and 72 h of exposure using both monolayers and 3D spheroids in human neuroblastoma SH-SY5Y cells. Furthermore, CPA toxicokinetics was evaluated using in silico models. Cytotoxicity increased dose- and time-dependently, as shown by the MTT assay. The lowest CPA IC₅₀ values were found in the monolayer study compared to the 3D spheroids at all exposure times (24 h: 864.01 vs. 1132; 48 h: 437 vs. 1069; 72 h: 392 vs. 567 nM). The CPA exposure on SH-SY5Y spheroid organization and morphology was also studied. Morphological changes, including spheroid disaggregation, were observed after mycotoxin exposure. The in silico methods, SwissADME and admetSAR, were used for short and full ADMEt profiles of CPA. The ADMEt predictive profile shows high gastrointestinal absorption and ability to penetrate the blood–brain barrier. Including in silico studies emphasizes the comprehensive approach to understanding mycotoxin toxicity and risk assessment. By combining in vitro 3D spheroid models with computational simulations, this study aims to provide a holistic perspective on the effects of CPA, enhancing the accuracy and relevance of our findings. Full article

(This article belongs to the Special Issue Toxins: 15th Anniversary)

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15 pages, 2138 KB

Open AccessEditor’s ChoiceArticle

Machine Learning Framework for Conotoxin Class and Molecular Target Prediction

by Duc P. Truong, Lyman K. Monroe, Robert F. Williams and Hau B. Nguyen

Toxins 2024, 16(11), 475; https://doi.org/10.3390/toxins16110475 - 3 Nov 2024

Cited by 2 | Viewed by 2743

Abstract

Conotoxins are small and highly potent neurotoxic peptides derived from the venom of marine cone snails which have captured the interest of the scientific community due to their pharmacological potential. These toxins display significant sequence and structure diversity, which results in a wide [...] Read more.

Conotoxins are small and highly potent neurotoxic peptides derived from the venom of marine cone snails which have captured the interest of the scientific community due to their pharmacological potential. These toxins display significant sequence and structure diversity, which results in a wide range of specificities for several different ion channels and receptors. Despite the recognized importance of these compounds, our ability to determine their binding targets and toxicities remains a significant challenge. Predicting the target receptors of conotoxins, based solely on their amino acid sequence, remains a challenge due to the intricate relationships between structure, function, target specificity, and the significant conformational heterogeneity observed in conotoxins with the same primary sequence. We have previously demonstrated that the inclusion of post-translational modifications, collisional cross sections values, and other structural features, when added to the standard primary sequence features, improves the prediction accuracy of conotoxins against non-toxic and other toxic peptides across varied datasets and several different commonly used machine learning classifiers. Here, we present the effects of these features on conotoxin class and molecular target predictions, in particular, predicting conotoxins that bind to nicotinic acetylcholine receptors (nAChRs). We also demonstrate the use of the Synthetic Minority Oversampling Technique (SMOTE)-Tomek in balancing the datasets while simultaneously making the different classes more distinct by reducing the number of ambiguous samples which nearly overlap between the classes. In predicting the alpha, mu, and omega conotoxin classes, the SMOTE-Tomek PCA PLR model, using the combination of the SS and P feature sets establishes the best performance with an overall accuracy (OA) of 95.95%, with an average accuracy (AA) of 93.04%, and an f1 score of 0.959. Using this model, we obtained sensitivities of 98.98%, 89.66%, and 90.48% when predicting alpha, mu, and omega conotoxin classes, respectively. Similarly, in predicting conotoxins that bind to nAChRs, the SMOTE-Tomek PCA SVM model, which used the collisional cross sections (CCSs) and the P feature sets, demonstrated the highest performance with 91.3% OA, 91.32% AA, and an f1 score of 0.9131. The sensitivity when predicting conotoxins that bind to nAChRs is 91.46% with a 91.18% sensitivity when predicting conotoxins that do not bind to nAChRs. Full article

(This article belongs to the Special Issue Conotoxins: Evolution, Classifications and Targets)

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15 pages, 4082 KB

Open AccessEditor’s ChoiceArticle

From Venom to Vein: Factor VII Activation as a Major Pathophysiological Target for Procoagulant Australian Elapid Snake Venoms

by Uthpala Chandrasekara, Abhinandan Chowdhury, Lorenzo Seneci, Christina N. Zdenek, Nathan Dunstan and Bryan G. Fry

Toxins 2024, 16(10), 430; https://doi.org/10.3390/toxins16100430 - 6 Oct 2024

Cited by 11 | Viewed by 3218

Abstract

Australian elapid snake venoms are uniquely procoagulant, utilizing blood clotting enzyme Factor Xa (FXa) as a toxin, which evolved as a basal trait in this clade. The subsequent recruitment of Factor Va (FVa) as a toxin occurred in the last common ancestor of [...] Read more.

Australian elapid snake venoms are uniquely procoagulant, utilizing blood clotting enzyme Factor Xa (FXa) as a toxin, which evolved as a basal trait in this clade. The subsequent recruitment of Factor Va (FVa) as a toxin occurred in the last common ancestor of taipans (Oxyuranus species) and brown snakes (Pseudonaja species). Factor II (prothrombin) activation has been stated as the primary mechanism for the lethal coagulopathy, but this hypothesis has never been tested. The additional activation of Factor VII (FVII) by Oxyuranus/Pseudonaja venoms has historically been considered as a minor, unimportant novelty. This study aimed to investigate the significance of toxic FVII activation relative to prothrombin activation by testing a wide taxonomical range of Australian elapid species with procoagulant venoms. The activation of FVII or prothrombin, with and without the Factor Va as a cofactor, was assessed, along with the structural changes involved in these processes. All procoagulant species could activate FVII, establishing this as a basal trait. In contrast, only some lineages could activate prothrombin, indicating that this is a derived trait. For species able to activate both zymogens, Factor VII was consistently more strongly activated than prothrombin. FVa was revealed as an essential cofactor for FVII activation, a mechanism previously undocumented. Species lacking FVa in their venom utilized endogenous plasma FVa to exert this activity. The ability of the human FXa:FVa complex to activate FVII was also revealed as a new feedback loop in the endogenous clotting cascade. Toxin sequence analyses identified structural changes essential for the derived trait of prothrombin activation. This study presents a paradigm shift in understanding how elapid venoms activate coagulation factors, highlighting the critical role of FVII activation in the pathophysiological effects upon the coagulation cascade produced by Australian elapid snake venoms. It also documented the novel use of Factor Va as a cofactor for FVII activation for both venom and endogenous forms of FXa. These findings are crucial for developing better antivenoms and treatments for snakebite victims and have broader implications for drug design and the treatment of coagulation disorders. The research also advances the evolutionary biology knowledge of snake venoms. Full article

(This article belongs to the Special Issue Animal Venoms: Unraveling the Molecular Complexity (2nd Edition))

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15 pages, 6767 KB

Open AccessEditor’s ChoiceArticle

Preparation of Soybean Dreg-Based Biochar@TiO₂ Composites and the Photocatalytic Degradation of Aflatoxin B₁ Exposed to Simulated Sunlight Irradiation

by Jian Zhang, Zhiwei Ying, He Li, Xinqi Liu, Dongge Ma and Hailong Yu

Toxins 2024, 16(10), 429; https://doi.org/10.3390/toxins16100429 - 5 Oct 2024

Cited by 15 | Viewed by 4733

Abstract

Aflatoxin B₁ (AFB₁) is a highly toxic carcinogen severely harmful to humans and animals. This study fabricated SDB-6-K-9@TiO₂ composites via the hydrothermal synthesis method to reduce AFB₁. The structural characterization results of the photocatalytic composites showed that [...] Read more.

Aflatoxin B₁ (AFB₁) is a highly toxic carcinogen severely harmful to humans and animals. This study fabricated SDB-6-K-9@TiO₂ composites via the hydrothermal synthesis method to reduce AFB₁. The structural characterization results of the photocatalytic composites showed that TiO₂ was successfully loaded onto SDB-6-K-9. The different photocatalytic degradation conditions, photocatalyst kinetics, recycling performance, and photocatalytic degradation mechanism were investigated. Photocatalysis with 6 mg of 4%SDB-6-K-9@TiO₂ in a 100 μg/mL AFB₁ solution presented a reduction of over 95%, exhibiting excellent performance, high stability, and reusability even after five cycles of photocatalytic experiments. Active species trapping experiments confirmed that holes (h⁺) played the most critical role. After structural analysis and identification of the photocatalytic degradation products, the photodegradation path and photocatalytic oxidation mechanism of 4%SDB-6-K-9@TiO₂ were postulated. The results show a new way to improve TiO₂’s photocatalytic performance, providing a certain theoretical basis for the effective AFB₁ reduction. Full article

(This article belongs to the Collection Aflatoxins)

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20 pages, 3372 KB

Open AccessEditor’s ChoiceArticle

Salinity as an Abiotic Stressor for Eliciting Bioactive Compounds in Marine Microalgae

by Adrián Macías-de la Rosa, Lorenzo López-Rosales, Antonio Contreras-Gómez, Asterio Sánchez-Mirón, Francisco García-Camacho and María del Carmen Cerón-García

Toxins 2024, 16(10), 425; https://doi.org/10.3390/toxins16100425 - 1 Oct 2024

Cited by 18 | Viewed by 7868

Abstract

This study investigated the impact of culture medium salinity (5–50 PSU) on the growth and maximum photochemical yield of photosystem II (Fv/Fm) and the composition of carotenoids, fatty acids, and bioactive substances in three marine microalgae (Chrysochromulina rotalis [...] Read more.

This study investigated the impact of culture medium salinity (5–50 PSU) on the growth and maximum photochemical yield of photosystem II (Fv/Fm) and the composition of carotenoids, fatty acids, and bioactive substances in three marine microalgae (Chrysochromulina rotalis, Amphidinium carterae, and Heterosigma akashiwo). The microalgae were photoautotrophically cultured in discontinuous mode in a single stage (S1) and a two-stage culture with salt shock (S2). A growth model was developed to link biomass productivity with salinity for each species. C. rotalis achieved a maximum biomass productivity (Pmax) of 15.85 ± 0.32 mg·L⁻¹·day⁻¹ in S1 and 16.12 ± 0.13 mg·L⁻¹·day⁻¹ in S2. The salt shock in S2 notably enhanced carotenoid production, particularly in C. rotalis and H. akashiwo, where fucoxanthin was the main carotenoid, while peridinin dominated in A. carterae. H. akashiwo also exhibited increased fatty acid productivity in S2. Salinity changes affected the proportions of saturated, monounsaturated, and polyunsaturated fatty acids in all three species. Additionally, hyposaline conditions boosted the production of haemolytic substances in A. carterae and C. rotalis. Full article

(This article belongs to the Special Issue Advances in Microalgae Toxins: Production, Detection, and Application 2.0)

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10 pages, 756 KB

Open AccessEditor’s ChoiceArticle

Comparative Effects of Acetate- and Citrate-Based Dialysates on Dialysis Dose and Protein-Bound Uremic Toxins in Hemodiafiltration Patients: Exploring the Impact of Calcium and Magnesium Concentrations

by Diana Rodríguez-Espinosa, Elena Cuadrado-Payán, Naira Rico, Mercè Torra, Rosa María Fernández, Miquel Gómez, Laura Morantes, Gregori Casals, Maria Rodriguez-Garcia, Francisco Maduell and José Jesús Broseta

Toxins 2024, 16(10), 426; https://doi.org/10.3390/toxins16100426 - 1 Oct 2024

Cited by 3 | Viewed by 3006

Abstract

Modern hemodialysis employs weak acids as buffers to prevent bicarbonate precipitation with calcium or magnesium. Acetate, the most used acid, is linked to chronic inflammation and poor dialysis tolerance. Citrate has emerged as a potential alternative, though its effect on dialysis efficiency is [...] Read more.

Modern hemodialysis employs weak acids as buffers to prevent bicarbonate precipitation with calcium or magnesium. Acetate, the most used acid, is linked to chronic inflammation and poor dialysis tolerance. Citrate has emerged as a potential alternative, though its effect on dialysis efficiency is not clear. This study aims to compare the efficacy of acetate- and citrate-based dialysates, focusing on protein-bound uremic toxins and dialysis doses. This single-center prospective crossover study includes prevalent patients participating in a thrice-weekly online hemodiafiltration program. Four dialysates were tested: two acetate-based (1.25 and 1.5 mmol/L calcium) and two citrate-based (1.5 mmol/L calcium with 0.5 and 0.75 mmol/L magnesium). Pre- and post-dialysis blood samples of eighteen patients were analyzed for urea, creatinine, p-cresyl sulfate, indoxyl sulfate, and albumin. Statistical significance was assessed using paired t-tests and repeated measures of ANOVA. There were no significant differences in dialysis dose (Kt), urea, creatinine, or indoxyl sulfate reduction ratios between acetate- and citrate-based dialysates. However, a significant decrease in the reduction ratio of p-cresyl sulfate was observed with the acetate dialysate containing 1.25 mmol/L calcium and the citrate dialysate with 0.5 mmol/L magnesium compared to the acetate dialysate containing 1.5 mmol/L calcium and the citrate dialysate with 0.75 mmol/L magnesium (51.56 ± 4.75 and 53.02 ± 4.52 vs. 65.25 ± 3.38 and 58.66 ± 4.16, p 0.007). No differences in dialysis dose were found between acetate- and citrate-based dialysates. However, citrate dialysates with lower calcium and magnesium concentrations may reduce the albumin displacement of p-cresyl sulfate. Further studies are needed to understand the observed differences and optimize the dialysate composition for the better clearance of protein-bound uremic toxins. Full article

(This article belongs to the Section Uremic Toxins)

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21 pages, 4548 KB

Open AccessEditor’s ChoiceArticle

Exposure of Cattle Breeding Herds to Naturally Co-Contaminated Zearalenone and Deoxynivalenol: The Relevance of a Urinary Mycotoxin Monitoring System for Herd Health and Food Safety

by Oky Setyo Widodo, Seiichi Uno, Emiko Kokushi, Osamu Yamato, M. Fariz Fadillah Mardianto, Urara Shinya, Yuto Kano, Chiho Kawashima, Yasuo Fushimi, Tetsushi Ono, Masayasu Taniguchi and Mitsuhiro Takagi

Toxins 2024, 16(9), 402; https://doi.org/10.3390/toxins16090402 - 18 Sep 2024

Cited by 6 | Viewed by 3158

Abstract

The widespread presence of Fusarium mycotoxins in animal feed is a global issue, not only for the health of livestock but also for ensure the safety of food as an end product. High concentrations of zearalenone (ZEN) and deoxynivalenol (DON) have been detected [...] Read more.

The widespread presence of Fusarium mycotoxins in animal feed is a global issue, not only for the health of livestock but also for ensure the safety of food as an end product. High concentrations of zearalenone (ZEN) and deoxynivalenol (DON) have been detected in the diets of Japanese Black (JB) and Holstein Friesian (HF) breeding herds. Consequently, we monitored serum biochemical parameters over a long time in both herds, focusing on anti-Müllerian hormone (AMH) levels and acute-phase inflammation. Additionally, urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) and progesterone levels were measured in the HF herd. The JB herd, a ZEN-dominant model with low DON contamination, demonstrated ZEN levels that exceeded the Japanese limit in the purchased total mixed rations (TMR). Conversely, the HF herd, which primary consumes DON-dominant feed with low ZEN contamination, had high DON levels in the dent corn silage. Specifically, the JB herd’s TMR contained 1.79 mg/kg ZEN and 0.58 mg/kg DON, whereas the HF herd’s silage had 15.3 mg/kg DON (dried sample) and 0.1 mg/kg ZEN. Enzyme-linked immunoassay were used to measure urinary ZEN-DON levels following confirmation through liquid chromatography-tandem mass spectrometry. Urinary ZEN-DON levels measured were significantly correlated (p < 0.05, r > 0.6) in both herds. In the HF herd, AMH levels increased (p = 0.01) and serum amyloid A (SAA) levels decreased (p = 0.02) when contaminated and at the end of the monitoring period. Additionally, urinary ZEN and DON levels were significantly correlated with SAA levels (ZEN: p = 0.00, r = 0.46; DON: p = 0.03, r = 0.33), with an increase in ZEN and DON levels resulting in higher SAA levels. The JB herd showed no significant differences. Additionally, in the HF herd, 8-OHdG/Cre levels increased significantly during major contamination periods (p < 0.05). Clinical data from the HF herd indicated an increase in mastitis cases and treatment rates during periods of major contamination. Abortion rates in the HF herd decreased from 22.9% (before monitoring) to 8.9% (during the high contamination period) and finally to 1% (at the end of the monitoring period), with corresponding increases in progesterone levels. ZEN-DON contamination adversely affects breeding cattle’s productivity, reproductive performance, and health. Therefore, monitoring urinary ZEN-DON is valuable for detecting contaminants and ensuring the safety of food products. Full article

(This article belongs to the Special Issue Toxicity Assessment, Rapid Detection and Control of Foodborne Mycotoxins: Advances and Prospects)

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17 pages, 3157 KB

Open AccessEditor’s ChoiceArticle

Caffeic Acid Phenethyl Ester Administration Reduces Enterotoxigenic Bacteroides fragilis-Induced Colitis and Tumorigenesis

by Soonjae Hwang, Minjeong Jo, Ju-Eun Hong, Woo-Seung Kim, Da-Hye Kang, Sang-Hyeon Yoo, Kyungsu Kang and Ki-Jong Rhee

Toxins 2024, 16(9), 403; https://doi.org/10.3390/toxins16090403 - 18 Sep 2024

Cited by 7 | Viewed by 3180

Abstract

The human colonic commensal enterotoxigenic Bacteroides fragilis (ETBF) is associated with chronic colitis and colon cancer. ETBF colonization induces colitis via the Bacteroides fragilis toxin (BFT). BFT secreted by ETBF cause colon inflammation via E-cadherin cleavage/NF-κB signaling. ETBF promotes colon tumorigenesis via interleukin [...] Read more.

The human colonic commensal enterotoxigenic Bacteroides fragilis (ETBF) is associated with chronic colitis and colon cancer. ETBF colonization induces colitis via the Bacteroides fragilis toxin (BFT). BFT secreted by ETBF cause colon inflammation via E-cadherin cleavage/NF-κB signaling. ETBF promotes colon tumorigenesis via interleukin 17A (IL-17A)/CXCL-dependent inflammation, but its bioactive therapeutics in ETBF-promoted tumorigenesis remain unexplored. In the current study, we investigated the caffeic acid phenethyl ester (CAPE) in the murine model of ETBF colitis and tumorigenesis. In this study, we observed that CAPE treatment mitigated inflammation induced by ETBF in mice. Additionally, our findings indicate that CAPE treatment offers protective effects against ETBF-enhanced colon tumorigenesis in a mouse model of colitis-associated colon cancer induced by azoxymethane (AOM) and dextran sulfate sodium. Notably, the decrease in colon tumorigenesis following CAPE administration correlates with a reduction in the expression of IL-17A and CXCL1 in the gastrointestinal tract. The molecular mechanism for CAPE-induced protection against ETBF-mediated tumorigenesis is mediated by IL-17A/CXCL1, and by NF-κB activity in intestinal epithelial cells. Our findings indicate that CAPE may serve as a preventive agent against the development of ETBF-induced colitis and colorectal cancer (CRC). Full article

(This article belongs to the Section Bacterial Toxins)

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16 pages, 37816 KB

Open AccessEditor’s ChoiceArticle

Combinatory Effects of Acrylamide and Deoxynivalenol on In Vitro Cell Viability and Cytochrome P450 Enzymes of Human HepaRG Cells

by Julia Beisl, Kristina Jochum, Yi Chen, Elisabeth Varga and Doris Marko

Toxins 2024, 16(9), 389; https://doi.org/10.3390/toxins16090389 - 10 Sep 2024

Cited by 3 | Viewed by 2653

Abstract

Acrylamide (AA) can be formed during the thermal processing of carbohydrate-rich foods. Deoxynivalenol (DON), a mycotoxin produced by Fusarium spp., contaminates many cereal-based products. In addition to potential co-exposure through a mixed diet, co-occurrence of AA and DON in thermally processed cereal-based products [...] Read more.

Acrylamide (AA) can be formed during the thermal processing of carbohydrate-rich foods. Deoxynivalenol (DON), a mycotoxin produced by Fusarium spp., contaminates many cereal-based products. In addition to potential co-exposure through a mixed diet, co-occurrence of AA and DON in thermally processed cereal-based products is also likely, posing the question of combinatory toxicological effects. In the present study, the effects of AA (0.001–3 mM) and DON (0.1–30 µM) on the cytotoxicity, gene transcription, and expression of major cytochrome P450 (CYP) enzymes were investigated in differentiated human hepatic HepaRG cells. In the chosen ratios of AA–DON (10:1; 100:1), cytotoxicity was clearly driven by DON and no overadditive effects were observed. Using quantitative real-time PCR, about twofold enhanced transcript levels of CYP1A1 were observed at low DON concentrations (0.3 and 1 µM), reflected by an increase in CYP1A activity in the EROD assay. In contrast, CYP2E1 and CYP3A4 gene transcription decreased in a concentration-dependent manner after incubation with DON (0.01–0.3 µM). Nevertheless, confocal microscopy showed comparably constant protein levels. The present study provided no indication of an induction of CYP2E1 as a critical step in AA bioactivation by co-occurrence with DON. Taken together, the combination of AA and DON showed no clear physiologically relevant interaction in HepaRG cells. Full article

(This article belongs to the Special Issue Current Research on Mycotoxins in Food and Feed: From Detection and Unravelling of Toxicity to Control)

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14 pages, 2658 KB

Open AccessEditor’s ChoiceArticle

Mitigation of Deoxynivalenol (DON)- and Aflatoxin B1 (AFB1)-Induced Immune Dysfunction and Apoptosis in Mouse Spleen by Curcumin

by Azhar Muhmood, Jianxin Liu, Dandan Liu, Shuiping Liu, Mahmoud M. Azzam, Muhammad Bilawal Junaid, Lili Hou, Guannan Le and Kehe Huang

Toxins 2024, 16(8), 356; https://doi.org/10.3390/toxins16080356 - 13 Aug 2024

Cited by 13 | Viewed by 3251

Abstract

In the context of the potential immunomodulatory properties of curcumin in counteracting the detrimental effects of concurrent exposure to Deoxynivalenol (DON) and Aflatoxin B1 (AFB1), a comprehensive 28-days trial was conducted utilizing 60 randomly allocated mice divided into four groups. Administration of curcumin [...] Read more.

In the context of the potential immunomodulatory properties of curcumin in counteracting the detrimental effects of concurrent exposure to Deoxynivalenol (DON) and Aflatoxin B1 (AFB1), a comprehensive 28-days trial was conducted utilizing 60 randomly allocated mice divided into four groups. Administration of curcumin at a dosage of 5 mg/kg body weight in conjunction with DON at 0.1 mg/kg and AFB₁ at 0.01 mg/kg body weight was undertaken to assess its efficacy. Results indicated that curcumin intervention demonstrated mitigation of splenic structural damage, augmentation of serum immunoglobulin A (IgA) and immunoglobulin G (IgG) levels, elevation in T lymphocyte subset levels, and enhancement in the mRNA expression levels of pro-inflammatory cytokines TNF-α, IFN-γ, IL-2, and IL-6. Furthermore, curcumin exhibited a suppressive effect on apoptosis in mice, as evidenced by decreased activity of caspase-3 and caspase-9, reduced expression levels of pro-apoptotic markers Bax and Cytochrome-c (Cyt-c) at both the protein and mRNA levels, and the maintenance of a balanced expression ratio of mitochondrial apoptotic regulators Bax and Bcl-2. Collectively, these findings offer novel insights into the therapeutic promise of curcumin in mitigating immunosuppression and apoptotic events triggered by mycotoxin co-exposure. Full article

(This article belongs to the Section Mycotoxins)

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26 pages, 5099 KB

Open AccessEditor’s ChoiceArticle

Potential Ancestral Conoidean Toxins in the Venom Cocktail of the Carnivorous Snail Raphitoma purpurea (Montagu, 1803) (Neogastropoda: Raphitomidae)

by Giacomo Chiappa, Giulia Fassio, Maria Vittoria Modica and Marco Oliverio

Toxins 2024, 16(8), 348; https://doi.org/10.3390/toxins16080348 - 9 Aug 2024

Cited by 5 | Viewed by 3150

Abstract

Venomous marine gastropods of the superfamily Conoidea possess a rich arsenal of toxins, including neuroactive toxins. Venom adaptations might have played a fundamental role in the radiation of conoideans; nevertheless, there is still no knowledge about the venom of the most diversified family [...] Read more.

Venomous marine gastropods of the superfamily Conoidea possess a rich arsenal of toxins, including neuroactive toxins. Venom adaptations might have played a fundamental role in the radiation of conoideans; nevertheless, there is still no knowledge about the venom of the most diversified family of the group: Raphitomidae Bellardi, 1875. In this study, transcriptomes were produced from the carcase, salivary glands, and proximal and distal venom ducts of the northeastern Atlantic species Raphitoma purpurea (Montagu, 1803). Using a gut barcoding approach, we were also able to report, for the first time, molecular evidence of a vermivorous diet for the genus. Transcriptomic analyses revealed over a hundred putative venom components (PVC), including 69 neurotoxins. Twenty novel toxin families, including some with high levels of expansion, were discovered. No significant difference was observed between the distal and proximal venom duct secretions. Peptides related to cone snail toxins (Cerm06, Pgam02, and turritoxin) and other venom-related proteins (disulfide isomerase and elevenin) were retrieved from the salivary glands. These salivary venom components may constitute ancestral adaptations for venom production in conoideans. Although often neglected, salivary gland secretions are of extreme importance for understanding the evolutionary history of conoidean venom. Full article

(This article belongs to the Special Issue Structure, Function and Evolution of Conotoxins)

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11 pages, 3032 KB

Open AccessEditor’s ChoiceArticle

Evaluating a Venom-Bioinspired Peptide, NOR-1202, as an Antiepileptic Treatment in Male Mice Models

by Maria Varela Torres Quintanilha, Giovanna de Azevedo Mello Gobbo, Gabriela Beserra Pinheiro, Adolfo Carlos Barros de Souza, Luana Cristina Camargo and Marcia Renata Mortari

Toxins 2024, 16(8), 342; https://doi.org/10.3390/toxins16080342 - 5 Aug 2024

Cited by 1 | Viewed by 2207

Abstract

Epilepsy, a neurological disorder characterized by excessive neuronal activity and synchronized electrical discharges, ranks among the most prevalent global neurological conditions. Despite common use, antiepileptic drugs often result in adverse effects and lack effectiveness in controlling seizures in temporal lobe epilepsy (TLE) patients. [...] Read more.

Epilepsy, a neurological disorder characterized by excessive neuronal activity and synchronized electrical discharges, ranks among the most prevalent global neurological conditions. Despite common use, antiepileptic drugs often result in adverse effects and lack effectiveness in controlling seizures in temporal lobe epilepsy (TLE) patients. Recent research explored the potential of occidentalin-1202, a peptide inspired by Polybia occidentalis venom, in safeguarding Wistar rats from chemically induced seizures. The present study evaluated the new analog from occidentalin-1202 named NOR-1202 using acute and chronic pilocarpine-induced models and an acute kainic acid (KA) male mice model. NOR-1202 was administered through the intracerebroventricular (i.c.v.), subcutaneous, or intraperitoneal routes, with stereotaxic procedures for the i.c.v. injection. In the acute pilocarpine-induced model, NOR-1202 (i.c.v.) protected against generalized seizures and mortality but lacked systemic antiepileptic activity. In the KA model, it did not prevent generalized seizures but improved survival. In the chronic TLE model, NOR-1202′s ED₅₀ did not differ significantly from the epileptic or healthy groups regarding time spent in spontaneous recurrent seizures during the five-day treatment. However, the NOR-1202 group exhibited more seizures than the healthy group on the second day of treatment. In summary, NOR-1202 exhibits antiepileptic effects against chemoconvulsant-induced seizures, but no effect was observed when administered systemically. Full article

(This article belongs to the Special Issue From Animal Venoms to Solutions: In Honor of Professor Lourival D. Possani on the Occasion of His 85th Birthday)

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21 pages, 2525 KB

Open AccessEditor’s ChoiceArticle

A Novel Cytotoxic Mechanism for Triple-Negative Breast Cancer Cells Induced by the Type II Heat-Labile Enterotoxin LT-IIc through Ganglioside Ligation

by Natalie D. King-Lyons, Aryana S. Bhati, John C. Hu, Lorrie M. Mandell, Gautam N. Shenoy, Hugh J. Willison and Terry D. Connell

Toxins 2024, 16(7), 311; https://doi.org/10.3390/toxins16070311 - 11 Jul 2024

Viewed by 2840

Abstract

Triple-negative breast cancer (TNBC), which constitutes 10–20 percent of all breast cancers, is aggressive, has high metastatic potential, and carries a poor prognosis due to limited treatment options. LT-IIc, a member of the type II subfamily of ADP-ribosylating—heat-labile enterotoxins that bind to a [...] Read more.

Triple-negative breast cancer (TNBC), which constitutes 10–20 percent of all breast cancers, is aggressive, has high metastatic potential, and carries a poor prognosis due to limited treatment options. LT-IIc, a member of the type II subfamily of ADP-ribosylating—heat-labile enterotoxins that bind to a distinctive set of cell-surface ganglioside receptors—is cytotoxic toward TNBC cell lines, but has no cytotoxic activity for non-transformed breast epithelial cells. Here, primary TNBC cells, isolated from resected human tumors, showed an enhanced cytotoxic response specifically toward LT-IIc, in contrast to other enterotoxins that were tested. MDA-MB-231 cells, a model for TNBC, were used to evaluate potential mechanisms of cytotoxicity by LT-IIc, which induced elevated intracellular cAMP and stimulated the cAMP response element-binding protein (CREB) signaling pathway. To dissect the role of ADP-ribosylation, cAMP induction, and ganglioside ligation in the cytotoxic response, MDA-MB-231 cells were exposed to wild-type LT-IIc, the recombinant B-pentamer of LT-IIc that lacks the ADP-ribosylating A polypeptide, or mutants of LT-IIc with an enzymatically inactivated A1-domain. These experiments revealed that the ADP-ribosyltransferase activity of LT-IIc was nonessential for inducing the lethality of MDA-MB-231 cells. In contrast, a mutant LT-IIc with an altered ganglioside binding activity failed to trigger a cytotoxic response in MDA-MB-231 cells. Furthermore, the pharmacological inhibition of ganglioside expression protected MDA-MB-231 cells from the cytotoxic effects of LT-IIc. These data establish that ganglioside ligation, but not the induction of cAMP production nor ADP-ribosyltransferase activity, is essential to initiating the LT-IIc-dependent cell death of MDA-MB-231 cells. These experiments unveiled previously unknown properties of LT-IIc and gangliosides in signal transduction, offering the potential for the targeted treatment of TNBC, an option that is desperately needed. Full article

(This article belongs to the Section Bacterial Toxins)

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20 pages, 4165 KB

Open AccessEditor’s ChoiceArticle

Bioremediation of Aflatoxin B₁ by Meyerozyma guilliermondii AF01 in Peanut Meal via Solid-State Fermentation

by Wan Zhang, Changpo Sun, Wei Wang and Zhongjie Zhang

Toxins 2024, 16(7), 305; https://doi.org/10.3390/toxins16070305 - 4 Jul 2024

Cited by 7 | Viewed by 2869

Abstract

The use of microorganisms to manage aflatoxin contamination is a gentle and effective approach. The aim of this study was to test the removal of AFB₁ from AFB₁-contaminated peanut meal by a strain of Meyerozyma guilliermondii AF01 screened by the [...] Read more.

The use of microorganisms to manage aflatoxin contamination is a gentle and effective approach. The aim of this study was to test the removal of AFB₁ from AFB₁-contaminated peanut meal by a strain of Meyerozyma guilliermondii AF01 screened by the authors and to optimize the conditions of the biocontrol. A regression model with the removal ratio of AFB₁ as the response value was established by means of single-factor and response surface experiments. It was determined that the optimal conditions for the removal of AFB₁ from peanut meal by AF01 were 75 h at 29 °C under the natural pH, with an inoculum of 5.5%; the removal ratio of AFB₁ reached 69.31%. The results of simulating solid-state fermentation in production using shallow pans and fermentation bags showed that the removal ratio of AFB₁ was 68.85% and 70.31% in the scaled-up experiments, respectively. This indicated that AF01 had strong adaptability to the environment with facultative anaerobic fermentation detoxification ability. The removal ratio of AFB₁ showed a positive correlation with the growth of AF01, and there were no significant changes in the appearance and quality of the peanut meal after fermentation. This indicated that AF01 had the potential to be used in practical production. Full article

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10 pages, 538 KB

Open AccessEditor’s ChoiceArticle

Enhancing Botulinum Toxin Injection Precision: The Efficacy of a Single Cadaveric Ultrasound Training Intervention for Improved Anatomical Localization

by Camille Heslot, Omar Khan, Alexis Schnitzler, Chloe Haldane, Romain David and Rajiv Reebye

Toxins 2024, 16(7), 304; https://doi.org/10.3390/toxins16070304 - 2 Jul 2024

Cited by 9 | Viewed by 3641

Abstract

Ultrasound guidance can enhance existing landmark-based injection methods, even through a brief and single exposure during a cadaveric training course. A total of twelve participants were enrolled in this training program, comprising nine physical medicine and rehabilitation specialists, one pediatrician, and two physician [...] Read more.

Ultrasound guidance can enhance existing landmark-based injection methods, even through a brief and single exposure during a cadaveric training course. A total of twelve participants were enrolled in this training program, comprising nine physical medicine and rehabilitation specialists, one pediatrician, and two physician assistants. For each participant, one upper-limb muscle and one lower-limb muscle were randomly chosen from the preselected muscle group. Subsequently, participants were tasked with injecting both of their chosen cadaveric muscles with 1 mL of acrylic paint using a manual needle palpation technique, relying solely on their knowledge of anatomic landmarks. Participants then underwent a personalized, one-to-one ultrasound teaching session, lasting approximately five minutes, conducted by two highly experienced instructors. Following this instructive phase, participants were tasked with a second round of injections, targeting the same two muscles in the lower and upper limbs. However, this time, the injections were performed using anatomical landmarks and ultrasound guidance. To facilitate differentiation from the initial injections, a distinct color of acrylic paint was employed. When employing the anatomical landmark-based approach, the overall success rate for injections was 67%, with 16 out of 24 targeted muscles accurately injected. With the incorporation of ultrasound guidance, the success rate was 92%, precisely targeting 22 out of the 24 muscles under examination. There was an improvement in injection accuracy achievable through the integration of ultrasound guidance, even with minimal training exposure. Our single cadaveric ultra-sound training program contributes valuable insights to the utilization of ultrasound for anatomy training to help optimize the targeting of BoNT-A. Full article

(This article belongs to the Special Issue Uses of Botulinum Toxin Injection in Medicine)

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24 pages, 4815 KB

Open AccessFeature PaperEditor’s ChoiceArticle

The Clot Thickens: Differential Coagulotoxic and Cardiotoxic Activities of Anguimorpha Lizard Venoms

by James Dobson, Abhinandan Chowdhury, Jeremie Tai-A-Pin, Harold van der Ploeg, Amber Gillett and Bryan G. Fry

Toxins 2024, 16(6), 283; https://doi.org/10.3390/toxins16060283 - 20 Jun 2024

Cited by 12 | Viewed by 4047

Abstract

Despite their evolutionary novelty, lizard venoms are much less studied in comparison to the intense research on snake venoms. While the venoms of helodermatid lizards have long been assumed to be for defensive purposes, there is increasing evidence of toxic activities more useful [...] Read more.

Despite their evolutionary novelty, lizard venoms are much less studied in comparison to the intense research on snake venoms. While the venoms of helodermatid lizards have long been assumed to be for defensive purposes, there is increasing evidence of toxic activities more useful for predation than defence (such as paralytic neurotoxicity). This study aimed to ascertain the effects of Heloderma, Lanthanotus, and Varanus lizard venoms on the coagulation and cardiovascular systems. Anticoagulant toxicity was demonstrated for the Varanus species studied, with the venoms prolonging clotting times in human and bird plasma due to the destructive cleavage of fibrinogen. In contrast, thromboelastographic analyses on human and bird plasmas in this study demonstrated a procoagulant bioactivity for Heloderma venoms. A previous study on Heloderma venom using factor-depleted plasmas as a proxy model suggested a procoagulant factor was present that activated either Factor XI or Factor XII, but could not ascertain the precise target. Our activation studies using purified zymogens confirmed FXII activation. Comparisons of neonate and adult H. exasperatum, revealed the neonates to be more potent in the ability to activate FXII, being more similar to the venom of the smaller species H. suspectum than the adult H. exasperatum. This suggests potent FXII activation a basal trait in the genus, present in the small bodied last common ancestor. This also indicates an ontogenetic difference in prey preferences in the larger Heloderma species paralleing the change in venom biochemistry. In addition, as birds lack Factor XII, the ability to clot avian plasma suggested an additional procoagulant site of action, which was revealed to be the activation of Factor VII, with H. horridum being the most potent. This study also examined the effects upon the cardiovascular system, including the liberation of kinins from kininogen, which contributes to hypotension induction. This form of toxicity was previously described for Heloderma venoms, and was revealed in this study was to also be a pathophysiological effect of Lanthanotus and Varanus venoms. This suggests that this toxic activity was present in the venom of the last common ancestor of the anguimorph lizards, which is consistent with kallikrein enzymes being a shared toxin trait. This study therefore uncovered novel actions of anguimorph lizard venoms, not only contributing to the evolutionary biology body of knowledge but also revealing novel activities to mine for drug design lead compounds. Full article

(This article belongs to the Special Issue Venom Components Acting on the Hemostatic System: Structural and Mechanistic Insights)

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15 pages, 3020 KB

Open AccessEditor’s ChoiceArticle

The Possible Role of Mycotoxins in the Pathogenesis of Endometrial Cancer

by Márkó Unicsovics, Zsófia Molnár, Miklós Mézes, Katalin Posta, György Nagyéri, Szabolcs Várbíró, Nándor Ács, Levente Sára and Zsuzsanna Szőke

Toxins 2024, 16(6), 236; https://doi.org/10.3390/toxins16060236 - 23 May 2024

Cited by 13 | Viewed by 4801

Abstract

Endometrial cancer is one of the most common cancer types among women. Many factors can contribute to the development of this disease, including environmental factors and, thus, eating habits. Our study aims to determine the levels of various mycotoxins and their metabolites in [...] Read more.

Endometrial cancer is one of the most common cancer types among women. Many factors can contribute to the development of this disease, including environmental factors and, thus, eating habits. Our study aims to determine the levels of various mycotoxins and their metabolites in the blood serum and endometrial tissue samples of participants with previously proven endometrial cancer and to find possible contributions to cancer development. In the cohort clinical trial, 52 participants aged between 44 and 86 were studied. The participants were divided into two groups: patients or matched controls. All patients had previously histologically diagnosed endometrial cancer. The cancer patients were divided into low-grade endometrioid and low- plus high-grade endometrioid groups. Controls had no history of endometrial malignancy or premalignancy. Blood serum and endometrial tissue samples were obtained from all study patients. We compared the concentrations of total Aflatoxins (Afs), Deoxynivalenol (DON), Ochratoxin-A (OTA), T2-toxin and HT2 toxin (T2/HT2 toxin), Zearalenone (ZEN), alpha-Zearalenol (α-ZOL), and Fumonisin B1 (FB1) in the serum and endometrium between the different study groups. As a result, we can see a significant correlation between the higher levels of Afs and zearalenone and the presence of endometrial cancer. In the case of Afs, DON, OTA, T2/HT2 toxins, ZEN, and alpha-ZOL, we measured higher endometrial concentrations than in serum. Considering the effect of mycotoxins and eating habits on cancer development, our results might lead to further research exploring the relationship between certain mycotoxins and endometrium cancer. Full article

(This article belongs to the Special Issue Advances in Contamination, Detection and Risk Assessment of Mycotoxins)

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10 pages, 4613 KB

Open AccessEditor’s ChoiceArticle

Borrelia burgdorferi 0755, a Novel Cytotoxin with Unknown Function in Lyme Disease

by Sam T. Donta

Toxins 2024, 16(6), 233; https://doi.org/10.3390/toxins16060233 - 21 May 2024

Cited by 1 | Viewed by 5054

Abstract

The pathophysiology of Lyme disease, especially in its persistent form, remains to be determined. As many of the neurologic symptoms are similar to those seen in other toxin-associated disorders, a hypothesis was generated that B. burgdorferi, the causative agent of Lyme disease, [...] Read more.

The pathophysiology of Lyme disease, especially in its persistent form, remains to be determined. As many of the neurologic symptoms are similar to those seen in other toxin-associated disorders, a hypothesis was generated that B. burgdorferi, the causative agent of Lyme disease, may produce a neurotoxin to account for some of the symptoms. Using primers against known conserved bacterial toxin groups, and PCR technology, a candidate neurotoxin was discovered. The purified protein was temporarily named BbTox, and was subsequently found to be identical to BB0755, a protein deduced from the genome sequence of B. burgdorferi that has been annotated as a Z ribonuclease. BbTox has cytotoxic activity against cells of neural origin in tissue culture. Its toxic activity appears to be directed against cytoskeletal elements, similar to that seen with toxins of Clostridioides difficile and Clostridioides botulinum, but differing from that of cholera and E. coli toxins, and other toxins. It remains to be determined whether BbTox has direct cytotoxic effects on neural or glial cells in vivo, or its activity is primarily that of a ribonuclease analogous to other bacterial ribonucleases that are involved in antibiotic tolerance remains to be determined. Full article

(This article belongs to the Section Bacterial Toxins)

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15 pages, 4425 KB

Open AccessEditor’s ChoiceArticle

Specificity of DNA ADP-Ribosylation Reversal by NADARs

by Bara Cihlova, Yang Lu, Andreja Mikoč, Marion Schuller and Ivan Ahel

Toxins 2024, 16(5), 208; https://doi.org/10.3390/toxins16050208 - 28 Apr 2024

Cited by 13 | Viewed by 5513

Abstract

Recent discoveries establish DNA and RNA as bona fide substrates for ADP-ribosylation. NADAR (“NAD- and ADP-ribose”-associated) enzymes reverse guanine ADP-ribosylation and serve as antitoxins in the DarT-NADAR operon. Although NADARs are widespread across prokaryotes, eukaryotes, and viruses, their specificity and broader physiological roles [...] Read more.

Recent discoveries establish DNA and RNA as bona fide substrates for ADP-ribosylation. NADAR (“NAD- and ADP-ribose”-associated) enzymes reverse guanine ADP-ribosylation and serve as antitoxins in the DarT-NADAR operon. Although NADARs are widespread across prokaryotes, eukaryotes, and viruses, their specificity and broader physiological roles remain poorly understood. Using phylogenetic and biochemical analyses, we further explore de-ADP-ribosylation activity and antitoxin functions of NADAR domains. We demonstrate that different subfamilies of NADAR proteins from representative E. coli strains and an E. coli-infecting phage retain biochemical activity while displaying specificity in providing protection from toxic guanine ADP-ribosylation in cells. Furthermore, we identify a myxobacterial enzyme within the YbiA subfamily that functions as an antitoxin for its associated DarT-unrelated ART toxin, which we termed YarT, thus presenting a hitherto uncharacterised ART-YbiA toxin–antitoxin pair. Our studies contribute to the burgeoning field of DNA ADP-ribosylation, supporting its physiological relevance within and beyond bacterial toxin–antitoxin systems. Notably, the specificity and confinement of NADARs to non-mammals infer their potential as highly specific targets for antimicrobial drugs with minimal off-target effects. Full article

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17 pages, 10327 KB

Open AccessEditor’s ChoiceArticle

Edodin: A New Type of Toxin from Shiitake Mushroom (Lentinula edodes) That Inactivates Mammalian Ribosomes

by Lucía Citores, Sara Ragucci, Claudia C. Gay, Rosita Russo, Angela Chambery, Antimo Di Maro, Rosario Iglesias and José M. Ferreras

Toxins 2024, 16(4), 185; https://doi.org/10.3390/toxins16040185 - 10 Apr 2024

Cited by 9 | Viewed by 4292

Abstract

Ribosome-inactivating proteins (RIPs) are a group of proteins with rRNA N-glycosylase activity that irreversibly inhibit protein synthesis and consequently cause cell death. Recently, an RIP called ledodin has been found in shiitake; it is cytotoxic, strongly inhibits protein synthesis, and shows rRNA N-glycosylase [...] Read more.

Ribosome-inactivating proteins (RIPs) are a group of proteins with rRNA N-glycosylase activity that irreversibly inhibit protein synthesis and consequently cause cell death. Recently, an RIP called ledodin has been found in shiitake; it is cytotoxic, strongly inhibits protein synthesis, and shows rRNA N-glycosylase activity. In this work, we isolated and characterized a 50 kDa cytotoxic protein from shiitake that we named edodin. Edodin inhibits protein synthesis in a mammalian cell-free system, but not in insect-, yeast-, and bacteria-derived systems. It exhibits rRNA N-glycosylase and DNA-nicking activities, which relate it to plant RIPs. It was also shown to be toxic to HeLa and COLO 320 cells. Its structure is not related to other RIPs found in plants, bacteria, or fungi, but, instead, it presents the characteristic structure of the fold type I of pyridoxal phosphate-dependent enzymes. Homologous sequences have been found in other fungi of the class Agaricomycetes; thus, edodin could be a new type of toxin present in many fungi, some of them edible, which makes them of great interest in health, both for their involvement in food safety and for their potential biomedical and biotechnological applications. Full article

(This article belongs to the Special Issue Biological Activities of Ribosome Inactivating Proteins II)

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11 pages, 4216 KB

Open AccessEditor’s ChoiceArticle

High-Voltage Toxin’Roll: Electrostatic Charge Repulsion as a Dynamic Venom Resistance Trait in Pythonid Snakes

by Uthpala Chandrasekara, Emilie M. Broussard, Darin R. Rokyta and Bryan G. Fry

Toxins 2024, 16(4), 176; https://doi.org/10.3390/toxins16040176 - 4 Apr 2024

Cited by 3 | Viewed by 3749

Abstract

The evolutionary interplay between predator and prey has significantly shaped the development of snake venom, a critical adaptation for subduing prey. This arms race has spurred the diversification of the components of venom and the corresponding emergence of resistance mechanisms in the prey [...] Read more.

The evolutionary interplay between predator and prey has significantly shaped the development of snake venom, a critical adaptation for subduing prey. This arms race has spurred the diversification of the components of venom and the corresponding emergence of resistance mechanisms in the prey and predators of venomous snakes. Our study investigates the molecular basis of venom resistance in pythons, focusing on electrostatic charge repulsion as a defense against α-neurotoxins binding to the alpha-1 subunit of the postsynaptic nicotinic acetylcholine receptor. Through phylogenetic and bioactivity analyses of orthosteric site sequences from various python species, we explore the prevalence and evolution of amino acid substitutions that confer resistance by electrostatic repulsion, which initially evolved in response to predatory pressure by Naja (cobra) species (which occurs across Africa and Asia). The small African species Python regius retains the two resistance-conferring lysines (positions 189 and 191) of the ancestral Python genus, conferring resistance to sympatric Naja venoms. This differed from the giant African species Python sebae, which has secondarily lost one of these lysines, potentially due to its rapid growth out of the prey size range of sympatric Naja species. In contrast, the two Asian species Python brongersmai (small) and Python bivittatus (giant) share an identical orthosteric site, which exhibits the highest degree of resistance, attributed to three lysine residues in the orthosteric sites. One of these lysines (at orthosteric position 195) evolved in the last common ancestor of these two species, which may reflect an adaptive response to increased predation pressures from the sympatric α-neurotoxic snake-eating genus Ophiophagus (King Cobras) in Asia. All these terrestrial Python species, however, were less neurotoxin-susceptible than pythons in other genera which have evolved under different predatory pressure as: the Asian species Malayopython reticulatus which is arboreal as neonates and juveniles before rapidly reaching sizes as terrestrial adults too large for sympatric Ophiophagus species to consider as prey; and the terrestrial Australian species Aspidites melanocephalus which occupies a niche, devoid of selection pressure from α-neurotoxic predatory snakes. Our findings underline the importance of positive selection in the evolution of venom resistance and suggest a complex evolutionary history involving both conserved traits and secondary evolution. This study enhances our understanding of the molecular adaptations that enable pythons to survive in environments laden with venomous threats and offers insights into the ongoing co-evolution between venomous snakes and their prey. Full article

(This article belongs to the Section Animal Venoms)

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20 pages, 912 KB

Open AccessEditor’s ChoiceArticle

Paradoxical Exception to Island Tameness: Increased Defensiveness in an Insular Population of Rattlesnakes

by William K. Hayes, Carl E. Person, Gerad A. Fox, Julie L. King, Erick Briggs and Eric C. K. Gren

Toxins 2024, 16(3), 157; https://doi.org/10.3390/toxins16030157 - 18 Mar 2024

Cited by 3 | Viewed by 6373

Abstract

Island tameness results largely from a lack of natural predators. Because some insular rattlesnake populations lack functional rattles, presumably the consequence of relaxed selection from reduced predation, we hypothesized that the Santa Catalina Island, California, USA, population of the southern Pacific rattlesnake ( [...] Read more.

Island tameness results largely from a lack of natural predators. Because some insular rattlesnake populations lack functional rattles, presumably the consequence of relaxed selection from reduced predation, we hypothesized that the Santa Catalina Island, California, USA, population of the southern Pacific rattlesnake (Crotalus helleri, which possesses a functional rattle), would exhibit a decrement in defensive behavior relative to their mainland counterparts. Contrary to our prediction, rattlesnakes from the island not only lacked tameness compared to mainland snakes, but instead exhibited measurably greater levels of defensiveness. Island snakes attempted to bite 4.7 times more frequently as we endeavored to secure them by hand, and required 2.1-fold more time to be pinned and captured. When induced to bite a beaker after being grasped, the island snakes also delivered 2.1-fold greater quantities of venom when controlling for body size. The additional venom resulted from 2.1-fold larger pulses of venom ejected from the fangs. We found no effects of duration in captivity (2–36 months), which suggests an absence of long-term habituation of antipredator behaviors. Breeding bird surveys and Christmas bird counts indicated reduced population densities of avian predators on Catalina compared to the mainland. However, historical estimates confirmed that populations of foxes and introduced mammalian predators (cats and pigs) and antagonists (herbivorous ungulates) substantially exceeded those on the mainland in recent centuries, and therefore best explain the paradoxically exaggerated defensive behaviors exhibited by Catalina’s rattlesnakes. These findings augment our understanding of anthropogenic effects on the behaviors of island animals and underscore how these effects can negatively affect human safety. Full article

(This article belongs to the Section Animal Venoms)

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15 pages, 1034 KB

Open AccessEditor’s ChoiceArticle

Association of Urinary Mycotoxins with Sperm Quality: A Case-Control Study in Southern Nigeria

by Friday Ebhodaghe Okonofua, Lorretta Favour Chizomam Ntoimo, Emmanuel Iyayi Unuabonah, Titus Afred Makudali Msagati, Oladiran Ayodeji, Michael Aziken, Akhere Omonkhua, Victor Ohenhen, Celestina Olafusi and Moses O. Alfred

Toxins 2024, 16(3), 119; https://doi.org/10.3390/toxins16030119 - 29 Feb 2024

Cited by 7 | Viewed by 4548

Abstract

The objective of this study was to determine the association between mycotoxins and the quality of spermatozoa in Nigeria. We designed a prospective case-control study involving 136 men diagnosed with reduced sperm count and quality in five infertility clinics in southwest Nigeria and [...] Read more.

The objective of this study was to determine the association between mycotoxins and the quality of spermatozoa in Nigeria. We designed a prospective case-control study involving 136 men diagnosed with reduced sperm count and quality in five infertility clinics in southwest Nigeria and 154 normal fertile controls. Sperm analysis was conducted in accordance with the recommendations of the World Health OrganizationWHO, while Liquid Chromatography–Mass Spectrometry was used to assay three metabolites of mycotoxins (zearalenone, ochratoxin A, and deoxyvinelol) in the urine samples of cases and controls. The data were analysed with descriptive statistics and non-parametric linear regression. The results showed no overall significant difference in levels of these metabolites between the cases and control groups. In contrast, higher levels of zearalenone and ochratoxin A significantly decreased sperm motility in the cases. Similarly, an increase in the level of ochratoxin A decreased sperm morphology in the unadjusted model in the cases. We conclude that exposure to mycotoxins reduces the quality of spermatozoa (motility and morphology) in Nigerian men but may have no effect on sperm count. Efforts to reduce the exposure of men to mycotoxins are important interventions to improve sperm quality and reduce the prevalence of male infertility in the country. Full article

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13 pages, 3878 KB

Open AccessEditor’s ChoiceArticle

Bee Venom Stimulates Growth Factor Release from Adipose-Derived Stem Cells to Promote Hair Growth

by Jung Hyun Kim, Tae Yoon Kim, Bonhyuk Goo and Yeoncheol Park

Toxins 2024, 16(2), 84; https://doi.org/10.3390/toxins16020084 - 4 Feb 2024

Cited by 8 | Viewed by 12361

Abstract

Limited evidence suggests that stimulating adipose-derived stem cells (ASCs) indirectly promotes hair growth. We examined whether bee venom (BV) activated ASCs and whether BV-induced hair growth was facilitated by enhanced growth factor release by ASCs. The induction of the telogen-to-anagen phase was studied [...] Read more.

Limited evidence suggests that stimulating adipose-derived stem cells (ASCs) indirectly promotes hair growth. We examined whether bee venom (BV) activated ASCs and whether BV-induced hair growth was facilitated by enhanced growth factor release by ASCs. The induction of the telogen-to-anagen phase was studied in mice. The underlying mechanism was investigated using organ cultures of mouse vibrissa hair follicles. When BV-treated ASCs were injected subcutaneously into mice, the telogen-to-anagen transition was accelerated and, by day 14, the hair weight increased. Quantitative polymerase chain reaction (qPCR) revealed that BV influenced the expression of several molecules, including growth factors, chemokines, channels, transcription factors, and enzymes. Western blot analysis was employed to verify the protein expression levels of extracellular-signal-regulated kinase (ERK) and phospho-ERK. Both the Boyden chamber experiment and scratch assay confirmed the upregulation of cell migration by BV. Additionally, ASCs secreted higher levels of growth factors after exposure to BV. Following BV therapy, the gene expression levels of alkaline phosphatase (ALP), fibroblast growth factor (FGF)-1 and 6, endothelial cell growth factor, and platelet-derived growth factor (PDGF)-C were upregulated. The findings of this study suggest that bee venom can potentially be utilized as an ASC-preconditioning agent for hair regeneration. Full article

(This article belongs to the Section Animal Venoms)

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14 pages, 1745 KB

Open AccessEditor’s ChoiceArticle

TDP-43 and Alzheimer’s Disease Pathology in the Brain of a Harbor Porpoise Exposed to the Cyanobacterial Toxin BMAA

by Susanna P. Garamszegi, Daniel J. Brzostowicki, Thomas M. Coyne, Regina T. Vontell and David A. Davis

Toxins 2024, 16(1), 42; https://doi.org/10.3390/toxins16010042 - 12 Jan 2024

Cited by 13 | Viewed by 5252

Abstract

Cetaceans are well-regarded as sentinels for toxin exposure. Emerging studies suggest that cetaceans can also develop neuropathological changes associated with neurodegenerative disease. The occurrence of neuropathology makes cetaceans an ideal species for examining the impact of marine toxins on the brain across the [...] Read more.

Cetaceans are well-regarded as sentinels for toxin exposure. Emerging studies suggest that cetaceans can also develop neuropathological changes associated with neurodegenerative disease. The occurrence of neuropathology makes cetaceans an ideal species for examining the impact of marine toxins on the brain across the lifespan. Here, we describe TAR DNA-binding protein 43 (TDP-43) proteinopathy and Alzheimer’s disease (AD) neuropathological changes in a beached harbor porpoise (Phocoena phocoena) that was exposed to a toxin produced by cyanobacteria called β-N-methylamino-L-alanine (BMAA). We found pathogenic TDP-43 cytoplasmic inclusions in neurons throughout the cerebral cortex, midbrain and brainstem. P62/sequestosome-1, responsible for the autophagy of misfolded proteins, was observed in the amygdala, hippocampus and frontal cortex. Genes implicated in AD and TDP-43 neuropathology such as APP and TARDBP were expressed in the brain. AD neuropathological changes such as amyloid-β plaques, neurofibrillary tangles, granulovacuolar degeneration and Hirano bodies were present in the hippocampus. These findings further support the development of progressive neurodegenerative disease in cetaceans and a potential causative link to cyanobacterial toxins. Climate change, nutrient pollution and industrial waste are increasing the frequency of harmful cyanobacterial blooms. Cyanotoxins like BMAA that are associated with neurodegenerative disease pose an increasing public health risk. Full article

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

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16 pages, 767 KB

Open AccessEditor’s ChoiceArticle

A Randomized, Double-Blind, Active Control, Multicenter, Phase 3 Study to Evaluate the Efficacy and Safety of Liztox^® versus Botox^® in Post-Stroke Upper Limb Spasticity

by Dong Hyun Ye, Min Ho Chun, Yoon Ghil Park, Nam-Jong Paik, Shi-Uk Lee, Seung Don Yoo and Deog Young Kim

Toxins 2023, 15(12), 697; https://doi.org/10.3390/toxins15120697 - 12 Dec 2023

Cited by 3 | Viewed by 6136

Abstract

Botulinum toxin type A (BTX-A) injection is a commonly used therapeutic intervention for upper limb spasticity in stroke patients. This study was designed as a randomized, active-drug-controlled, double-blind, multicenter, phase 3 clinical trial to evaluate the safety and efficacy of Liztox^® in [...] Read more.

Botulinum toxin type A (BTX-A) injection is a commonly used therapeutic intervention for upper limb spasticity in stroke patients. This study was designed as a randomized, active-drug-controlled, double-blind, multicenter, phase 3 clinical trial to evaluate the safety and efficacy of Liztox^® in comparison to onabotulinum toxin A (Botox^®) for individuals with post-stroke upper limb spasticity. The primary outcome was the alteration in wrist flexor muscle tone from the initial assessment to the fourth week, evaluated using the modified Ashworth scale (MAS). Secondary outcomes included MAS score changes for the wrist at weeks 8 and 12 from baseline; MAS score changes for finger and elbow flexors; and changes in the Disability Assessment Scale (DAS), Subject’s Global Assessment (SGA), the Investigator’s Global Assessment (IGA), and Caregiver Burden Scale (CBS) at weeks 4, 8, and 12 from baseline. The MAS score for wrist flexor spasticity decreased by −1.14 ± 0.59 in the Liztox^® group and −1.22 ± 0.59 in the Botox^® group from baseline to week 4. The difference [97.5% confidence interval (CI)] between the test and control groups was 0.08 [−∞, 0.26], confirming the non-inferiority of the test group compared to the control group. Furthermore, there were consistent improvements in the IGA, SGA, and CBS scores across all assessment intervals, with no statistically significant variances detected between the two groups. No safety-related concerns were reported during the study. In conclusion, Liztox^® injection proved to be a secure and efficacious intervention for managing upper extremity spasticity in post-stroke patients. Full article

(This article belongs to the Special Issue Botulinum Toxins Injections for Hypertonic Limbs—Is It So Effective?)

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17 pages, 1783 KB

Open AccessEditor’s ChoiceArticle

Comparative Analysis of Secondary Metabolites Produced by Ascochyta fabae under In Vitro Conditions and Their Phytotoxicity on the Primary Host, Vicia faba, and Related Legume Crops

by Eleonora Barilli, Pierluigi Reveglia, Francisco J. Agudo-Jurado, Vanessa Cañete García, Alessio Cimmino, Antonio Evidente and Diego Rubiales

Toxins 2023, 15(12), 693; https://doi.org/10.3390/toxins15120693 - 9 Dec 2023

Cited by 2 | Viewed by 3804

Abstract

Ascochyta blight, caused by Ascochyta fabae, poses a significant threat to faba bean and other legumes worldwide. Necrotic lesions on stems, leaves, and pods characterize the disease. Given the economic impact of this pathogen and the potential involvement of secondary metabolites in [...] Read more.

Ascochyta blight, caused by Ascochyta fabae, poses a significant threat to faba bean and other legumes worldwide. Necrotic lesions on stems, leaves, and pods characterize the disease. Given the economic impact of this pathogen and the potential involvement of secondary metabolites in symptom development, a study was conducted to investigate the fungus’s ability to produce bioactive metabolites that might contribute to its pathogenicity. For this investigation, the fungus was cultured in three substrates (Czapek-Dox, PDB, and rice). The produced metabolites were analyzed by NMR and LC-HRMS methods, resulting in the dereplication of seven metabolites, which varied with the cultural substrates. Ascochlorin, ascofuranol, and (R)-mevalonolactone were isolated from the Czapek-Dox extract; ascosalipyrone, benzoic acid, and tyrosol from the PDB extract; and ascosalitoxin and ascosalipyrone from the rice extract. The phytotoxicity of the pure metabolites was assessed at different concentrations on their primary hosts and related legumes. The fungal exudates displayed varying degrees of phytotoxicity, with the Czapek-Dox medium’s exudate exhibiting the highest activity across almost all legumes tested. The species belonging to the genus Vicia spp. were the most susceptible, with faba bean being susceptible to all metabolites, at least at the highest concentration tested, as expected. In particular, ascosalitoxin and benzoic acid were the most phytotoxic in the tested condition and, as a consequence, expected to play an important role on necrosis’s appearance. Full article

(This article belongs to the Special Issue Biological Activities and Potential Applications of Phytotoxins)

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17 pages, 1087 KB

Open AccessEditor’s ChoiceArticle

Turmeric Powder Counteracts Oxidative Stress and Reduces AFB1 Content in the Liver of Broilers Exposed to the EU Maximum Levels of the Mycotoxin

by Neenu Amminikutty, Veronica Spalenza, Watanya Jarriyawattanachaikul, Paola Badino, Maria Teresa Capucchio, Elena Colombino, Achille Schiavone, Donato Greco, Vito D’Ascanio, Giuseppina Avantaggiato, Sihem Dabbou, Carlo Nebbia and Flavia Girolami

Toxins 2023, 15(12), 687; https://doi.org/10.3390/toxins15120687 - 7 Dec 2023

Cited by 26 | Viewed by 4809

Abstract

The most frequent adverse effects of AFB1 in chicken are low performance, the depression of the immune system, and a reduced quality of both eggs and meat, leading to economic losses. Since oxidative stress plays a major role in AFB1 toxicity, natural products [...] Read more.

The most frequent adverse effects of AFB1 in chicken are low performance, the depression of the immune system, and a reduced quality of both eggs and meat, leading to economic losses. Since oxidative stress plays a major role in AFB1 toxicity, natural products are increasingly being used as an alternative to mineral binders to tackle AFB1 toxicosis in farm animals. In this study, an in vivo trial was performed by exposing broilers for 10 days to AFB1 at dietary concentrations approaching the maximum limits set by the EU (0.02 mg/kg feed) in the presence or absence of turmeric powder (TP) (included in the feed at 400 mg/kg). The aims were to evaluate (i) the effects of AFB1 on lipid peroxidation, antioxidant parameters, histology, and the expression of drug transporters and biotransformation enzymes in the liver; (ii) the hepatic accumulation of AFB1 and its main metabolites (assessed using an in-house-validated HPLC-FLD method); (iii) the possible modulation of the above parameters elicited by TP. Broilers exposed to AFB1 alone displayed a significant increase in lipid peroxidation in the liver, which was completely reverted by the concomitant administration of TP. Although no changes in glutathione levels and antioxidant enzyme activities were detected in any treatment group, AFB1 significantly upregulated and downregulated the mRNA expression of CYP2A6 and Nrf2, respectively. TP counteracted such negative effects and increased the hepatic gene expression of selected antioxidant enzymes (i.e., CAT and SOD2) and drug transporters (i.e., ABCG2), which were further enhanced in combination with AFB1. Moreover, both AFB1 and TP increased the mRNA levels of ABCC2 and ABCG2 in the duodenum. The latter changes might be implicated in the decrease in hepatic AFB1 to undetectable levels (<LOD) in the TP supplemented group. Overall, our findings further support the use of TP as an effective feeding strategy to prevent AFB1-related adverse effects in broilers. Full article

(This article belongs to the Special Issue Mycotoxins in Farm Animals: Health Disorders and Preventive Strategies)

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16 pages, 2712 KB

Open AccessEditor’s ChoiceCommunication

ADDovenom: Thermostable Protein-Based ADDomer Nanoparticles as New Therapeutics for Snakebite Envenoming

by Stefanie K. Menzies, Raquel Arinto-Garcia, Fernanda Gobbi Amorim, Iara Aimê Cardoso, Camille Abada, Thomas Crasset, Fabien Durbesson, Rebecca J. Edge, Priscila El-Kazzi, Sophie Hall, Damien Redureau, Richard Stenner, Johara Boldrini-França, Huan Sun, António Roldão, Paula M. Alves, Robert A. Harrison, Renaud Vincentelli, Imre Berger, Loïc Quinton, Nicholas R. Casewell and Christiane Schaffitzel Show full author list Hide full author list

Toxins 2023, 15(12), 673; https://doi.org/10.3390/toxins15120673 - 28 Nov 2023

Cited by 4 | Viewed by 4936

Abstract

Snakebite envenoming can be a life-threatening medical emergency that requires prompt medical intervention to neutralise the effects of venom toxins. Each year up to 138,000 people die from snakebites and threefold more victims suffer life-altering disabilities. The current treatment of snakebite relies solely [...] Read more.

Snakebite envenoming can be a life-threatening medical emergency that requires prompt medical intervention to neutralise the effects of venom toxins. Each year up to 138,000 people die from snakebites and threefold more victims suffer life-altering disabilities. The current treatment of snakebite relies solely on antivenom—polyclonal antibodies isolated from the plasma of hyperimmunised animals—which is associated with numerous deficiencies. The ADDovenom project seeks to deliver a novel snakebite therapy, through the use of an innovative protein-based scaffold as a next-generation antivenom. The ADDomer is a megadalton-sized, thermostable synthetic nanoparticle derived from the adenovirus penton base protein; it has 60 high-avidity binding sites to neutralise venom toxins. Here, we outline our experimental strategies to achieve this goal using state-of-the-art protein engineering, expression technology and mass spectrometry, as well as in vitro and in vivo venom neutralisation assays. We anticipate that the approaches described here will produce antivenom with unparalleled efficacy, safety and affordability. Full article

(This article belongs to the Section Animal Venoms)

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20 pages, 10566 KB

Open AccessEditor’s ChoiceArticle

An Assessment of Potential Threats to Human Health from Algae Blooms in the Indian River Lagoon (USA) 2018–2021: Unique Patterns of Cytotoxicity Associated with Toxins

by Esther A. Guzmán, Tara A. Peterson, Priscilla L. Winder, Kirstie T. Francis, Malcolm McFarland, Jill C. Roberts, Jennifer Sandle and Amy E. Wright

Toxins 2023, 15(11), 664; https://doi.org/10.3390/toxins15110664 - 17 Nov 2023

Cited by 4 | Viewed by 5630

Abstract

The Indian River Lagoon (IRL), a 156-mile-long estuary located on the eastern coast of Florida, experiences phytoplankton bloom events due to increased seasonal temperatures coupled with anthropogenic impacts. This study aimed to gather data on the toxicity to human cells and to identify [...] Read more.

The Indian River Lagoon (IRL), a 156-mile-long estuary located on the eastern coast of Florida, experiences phytoplankton bloom events due to increased seasonal temperatures coupled with anthropogenic impacts. This study aimed to gather data on the toxicity to human cells and to identify secondary metabolites found in water samples collected in the IRL. Water samples from 20 sites of the IRL were collected during the wet and dry seasons over a three-year period. A panel of cell lines was used to test cytotoxicity. Hemagglutination, hemolysis, and inhibition of protein phosphatase 2A (PP2A) were also measured. Cytotoxic blooms were seen both in the south (Microcystis) and the north (Pyrodinium) of the IRL. Each toxin induced a consistent pattern of cytotoxicity in the panel of human cell lines assayed. During blooms, cytotoxicity due to a single type of toxin is obvious from this pattern. In the absence of blooms, the cytotoxicity seen reflected either a mixture of toxins or it was caused by an unidentified toxin. These observations suggest that other toxins with the potential to be harmful to human health may be present in the IRL. Moreover, the presence of toxins in the IRL is not always associated with blooms of known toxin-producing organisms. Full article

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

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11 pages, 1922 KB

Open AccessEditor’s ChoiceArticle

Synergistic Effects of Essential Oils and Organic Acids against Aspergillus flavus Contamination in Poultry Feed

by Tim Satterlee, Callie Megan McDonough, Scott E. Gold, Chongxiao Chen, Anthony E. Glenn and Anthony Pokoo-Aikins

Toxins 2023, 15(11), 635; https://doi.org/10.3390/toxins15110635 - 31 Oct 2023

Cited by 10 | Viewed by 4334

Abstract

Organic acids and essential oils are commonly used in the poultry industry as antimicrobials and for their beneficial effects on gut health, growth performance, and meat quality. A common postharvest storage fungal colonist, Aspergillus flavus, contaminates corn, the primary component of poultry [...] Read more.

Organic acids and essential oils are commonly used in the poultry industry as antimicrobials and for their beneficial effects on gut health, growth performance, and meat quality. A common postharvest storage fungal colonist, Aspergillus flavus, contaminates corn, the primary component of poultry feed, with the highly detrimental mycotoxin, aflatoxin. Aflatoxin adversely affects poultry feed intake, feed conversion efficiency, weight gain, egg production, fertility, hatchability, and poultry meat yield. Both organic acids and essential oils have been reported to inhibit the growth of A. flavus. Thus, we evaluated if the inhibitory synergy between combined essential oils (cinnamon, lemongrass, and oregano) and organic acids (acetic, butyric, and propionic) prevents A. flavus growth. The study confirmed that these compounds inhibit the growth of A. flavus and that synergistic interactions do occur between some of them. Overall, cinnamon oil was shown to have the highest synergy with all the organic acids tested, requiring 1000 µL/L air of cinnamon oil and 888 mg/kg of butyric acid to fully suppress A. flavus growth on corn kernels. With the strong synergism demonstrated, combining certain essential oils and organic acids offers a potentially effective natural method for controlling postharvest aflatoxin contamination in poultry feed. Full article

(This article belongs to the Section Mycotoxins)

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15 pages, 2959 KB

Open AccessEditor’s ChoiceArticle

Ensemble Machine Learning of Gradient Boosting (XGBoost, LightGBM, CatBoost) and Attention-Based CNN-LSTM for Harmful Algal Blooms Forecasting

by Jung Min Ahn, Jungwook Kim and Kyunghyun Kim

Toxins 2023, 15(10), 608; https://doi.org/10.3390/toxins15100608 - 10 Oct 2023

Cited by 174 | Viewed by 16538

Abstract

Harmful algal blooms (HABs) are a serious threat to ecosystems and human health. The accurate prediction of HABs is crucial for their proactive preparation and management. While mechanism-based numerical modeling, such as the Environmental Fluid Dynamics Code (EFDC), has been widely used in [...] Read more.

Harmful algal blooms (HABs) are a serious threat to ecosystems and human health. The accurate prediction of HABs is crucial for their proactive preparation and management. While mechanism-based numerical modeling, such as the Environmental Fluid Dynamics Code (EFDC), has been widely used in the past, the recent development of machine learning technology with data-based processing capabilities has opened up new possibilities for HABs prediction. In this study, we developed and evaluated two types of machine learning-based models for HABs prediction: Gradient Boosting models (XGBoost, LightGBM, CatBoost) and attention-based CNN-LSTM models. We used Bayesian optimization techniques for hyperparameter tuning, and applied bagging and stacking ensemble techniques to obtain the final prediction results. The final prediction result was derived by applying the optimal hyperparameter and bagging and stacking ensemble techniques, and the applicability of prediction to HABs was evaluated. When predicting HABs with an ensemble technique, it is judged that the overall prediction performance can be improved by complementing the advantages of each model and averaging errors such as overfitting of individual models. Our study highlights the potential of machine learning-based models for HABs prediction and emphasizes the need to incorporate the latest technology into this important field. Full article

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

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17 pages, 1968 KB

Open AccessEditor’s ChoiceArticle

Discovery of an Insect Neuroactive Helix Ring Peptide from Ant Venom

by Valentine Barassé, Laurence Jouvensal, Guillaume Boy, Arnaud Billet, Steven Ascoët, Benjamin Lefranc, Jérôme Leprince, Alain Dejean, Virginie Lacotte, Isabelle Rahioui, Catherine Sivignon, Karen Gaget, Mélanie Ribeiro Lopes, Federica Calevro, Pedro Da Silva, Karine Loth, Françoise Paquet, Michel Treilhou, Elsa Bonnafé and Axel Touchard

Toxins 2023, 15(10), 600; https://doi.org/10.3390/toxins15100600 - 5 Oct 2023

Cited by 12 | Viewed by 5111

Abstract

Ants are among the most abundant terrestrial invertebrate predators on Earth. To overwhelm their prey, they employ several remarkable behavioral, physiological, and biochemical innovations, including an effective paralytic venom. Ant venoms are thus cocktails of toxins finely tuned to disrupt the physiological systems [...] Read more.

Ants are among the most abundant terrestrial invertebrate predators on Earth. To overwhelm their prey, they employ several remarkable behavioral, physiological, and biochemical innovations, including an effective paralytic venom. Ant venoms are thus cocktails of toxins finely tuned to disrupt the physiological systems of insect prey. They have received little attention yet hold great promise for the discovery of novel insecticidal molecules. To identify insect-neurotoxins from ant venoms, we screened the paralytic activity on blowflies of nine synthetic peptides previously characterized in the venom of Tetramorium bicarinatum. We selected peptide U₁₁, a 34-amino acid peptide, for further insecticidal, structural, and pharmacological experiments. Insecticidal assays revealed that U₁₁ is one of the most paralytic peptides ever reported from ant venoms against blowflies and is also capable of paralyzing honeybees. An NMR spectroscopy of U₁₁ uncovered a unique scaffold, featuring a compact triangular ring helix structure stabilized by a single disulfide bond. Pharmacological assays using Drosophila S2 cells demonstrated that U₁₁ is not cytotoxic, but suggest that it may modulate potassium conductance, which structural data seem to corroborate and will be confirmed in a future extended pharmacological investigation. The results described in this paper demonstrate that ant venom is a promising reservoir for the discovery of neuroactive insecticidal peptides. Full article

(This article belongs to the Special Issue Ant Venom)

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17 pages, 1558 KB

Open AccessEditor’s ChoiceArticle

Use of Yeast Cell Wall Extract for Growing Pigs Consuming Feed Contaminated with Mycotoxins below or above Regulatory Guidelines: A Meta-Analysis with Meta-Regression

by Alexandra C. Weaver, Daniel M. Weaver, Nicholas Adams and Alexandros Yiannikouris

Toxins 2023, 15(10), 596; https://doi.org/10.3390/toxins15100596 - 3 Oct 2023

Cited by 4 | Viewed by 4493

Abstract

Using a random-effects meta-analysis, the performance of growing pigs under a mycotoxin challenge (MT) with or without supplementation of yeast cell wall extract (YCWE, Mycosorb^®, Alltech Inc.) was evaluated. Both MT and YCWE were also compared to animal controls not receiving [...] Read more.

Using a random-effects meta-analysis, the performance of growing pigs under a mycotoxin challenge (MT) with or without supplementation of yeast cell wall extract (YCWE, Mycosorb^®, Alltech Inc.) was evaluated. Both MT and YCWE were also compared to animal controls not receiving mycotoxins (CTRL). Meta-regression was used to further explore the impacts of MT at/below (category 1) or above (category 2) global regulatory guidelines. Following the screening, 23 suitable references (30 mycotoxin treatments) were used. Overall, MT lowered average daily gain (ADG, p < 0.001) and average daily feed intake (ADFI, p < 0.0001) from CTRL by −84 and −165 g, respectively. Inclusion of YCWE during mycotoxin challenges (YCWE+MT, average 2.1 kg/ton) tended to result in greater ADG (+17 g, p = 0.068) compared to MT treatments. The gain-to-feed ratio (G:F) was not impacted by MT or YCWE+MT. Further investigation by meta-regression revealed that pigs fed MT in category 1 had lower ADG (−78.5 g, p < 0.001) versus CTRL, while YCWE+MT had higher ADG (+48 g, p < 0.001) over MT and was similar to CTRL. The ADFI was not impacted, although YCWE+MT had ADFI values similar to the CTRL. In category 2, ADG and ADFI of pigs fed MT were lower than CTRL (−85.1 and −166 g, respectively, p < 0.0001), with a tendency for YCWE+MT to result in higher ADFI (+25.3 g, p = 0.062). In summary, the inclusion of YCWE provided benefits to performance during common mycotoxin challenge levels (at or below regulatory guidelines). Full article

(This article belongs to the Special Issue Current Research on Mycotoxins in Food and Feed: From Detection and Unravelling of Toxicity to Control)

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14 pages, 3931 KB

Open AccessEditor’s ChoiceArticle

Presence, Co-Occurrence, and Daily Intake Estimates of Aflatoxins and Fumonisins in Maize Consumed in Food-Insecure Regions of Western Honduras

by Luis Sabillón, Jackeline Alvarado, Alejandra Leiva, Rodrigo Mendoza, Raúl Espinal, John F. Leslie and Andréia Bianchini

Toxins 2023, 15(9), 559; https://doi.org/10.3390/toxins15090559 - 7 Sep 2023

Cited by 8 | Viewed by 3655

Abstract

Foodborne mycotoxins are a significant food safety risk in developing countries. Our objective was to determine the occurrence of and exposure levels to aflatoxins (AFs) and fumonisins (FBs) in maize intended for human and animal consumption in food-insecure regions of western Honduras. Total [...] Read more.

Foodborne mycotoxins are a significant food safety risk in developing countries. Our objective was to determine the occurrence of and exposure levels to aflatoxins (AFs) and fumonisins (FBs) in maize intended for human and animal consumption in food-insecure regions of western Honduras. Total AFs and FBs were quantified with a monoclonal antibody-based affinity spectrofluorimetric method. FBs were detected in 614/631 samples of maize destined for human consumption at 0.3 to 41 mg/kg (mean, 2.7 mg/kg). Of the 614 positive samples, 147 had FB levels exceeding the U.S. Food and Drug Administration (FDA) advisory threshold of 4.0 mg/kg. AFs were detected in 109/631 samples of maize for human consumption with concentrations between 1.0 and 490 µg/kg (mean, 10 µg/kg). AF levels in 34 samples exceeded the FDA regulatory limit (i.e., 20 µg/kg). The average probable daily intake of AFs in western Honduras ranged from 0 to 260 ng/kg body weight/day, and for FBs, the average probable daily intake ranged from 17 to 53 μg/kg body weight/day. AFs and FBs co-occurred in 106/631 samples with 60 samples containing both toxins at levels greater than the FDA regulatory levels. Samples of maize intended for animal feed had significantly higher AF (mean, 22 µg/kg) and FB (mean, 7.6 mg/kg) contamination levels than those observed in samples destined for human consumption. Thus, the maize supply chain in western Honduras is contaminated with mycotoxins at levels that pose health risks to both humans and livestock. More effective mycotoxin surveillance and implementation of effective mitigation strategies are needed to reduce mycotoxin contamination and exposure. Full article

(This article belongs to the Special Issue Occurrence and Determination of Mycotoxins)

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19 pages, 4401 KB

Open AccessEditor’s ChoiceArticle

Identification of Acid Hydrolysis Metabolites of the Pimelea Toxin Simplexin for Targeted UPLC-MS/MS Analysis

by Zhi Hung Loh, Natasha L. Hungerford, Diane Ouwerkerk, Athol V. Klieve and Mary T. Fletcher

Toxins 2023, 15(9), 551; https://doi.org/10.3390/toxins15090551 - 5 Sep 2023

Cited by 2 | Viewed by 3609

Abstract

Pimelea poisoning of cattle is a unique Australian toxic condition caused by the daphnane orthoester simplexin present in native Pimelea pasture plants. Rumen microorganisms have been proposed to metabolise simplexin by enzymatic reactions, likely at the orthoester and epoxide moieties of simplexin, but [...] Read more.

Pimelea poisoning of cattle is a unique Australian toxic condition caused by the daphnane orthoester simplexin present in native Pimelea pasture plants. Rumen microorganisms have been proposed to metabolise simplexin by enzymatic reactions, likely at the orthoester and epoxide moieties of simplexin, but a metabolic pathway has not been confirmed. This study aimed to investigate this metabolic pathway through the analysis of putative simplexin metabolites. Purified simplexin was hydrolysed with aqueous hydrochloric acid and sulfuric acid to produce target metabolites for UPLC-MS/MS analysis of fermentation fluid samples, bacterial isolate samples, and other biological samples. UPLC-MS/MS analysis identified predicted hydrolysed products from both acid hydrolysis procedures with MS breakdown of these putative products sharing high-resolution accurate mass (HRAM) fragmentation ions with simplexin. However, targeted UPLC-MS/MS analysis of the biological samples failed to detect the H₂SO₄ degradation products, suggesting that the rumen microorganisms were unable to produce similar simplexin degradation products at detectable levels, or that metabolites, once formed, were further metabolised. Overall, in vitro acid hydrolysis was able to hydrolyse simplexin at the orthoester and epoxide functionalities, but targeted UPLC-MS/MS analysis of biological samples did not detect any of the identified simplexin hydrolysis products. Full article

(This article belongs to the Special Issue Toxic Compounds from Poisonous Plants: Identification and Analytical Methods)

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16 pages, 14537 KB

Open AccessEditor’s ChoiceArticle

Intramuscular Bleeding and Formation of Microthrombi during Skeletal Muscle Damage Caused by a Snake Venom Metalloprotease and a Cardiotoxin

by Medha Sonavane, José R. Almeida, Elanchezhian Rajan, Harry F. Williams, Felix Townsend, Elizabeth Cornish, Robert D. Mitchell, Ketan Patel and Sakthivel Vaiyapuri

Toxins 2023, 15(9), 530; https://doi.org/10.3390/toxins15090530 - 28 Aug 2023

Cited by 12 | Viewed by 4996

Abstract

The interactions between specific snake venom toxins and muscle constituents are the major cause of severe muscle damage that often result in amputations and subsequent socioeconomic ramifications for snakebite victims and/or their families. Therefore, improving our understanding of venom-induced muscle damage and determining [...] Read more.

The interactions between specific snake venom toxins and muscle constituents are the major cause of severe muscle damage that often result in amputations and subsequent socioeconomic ramifications for snakebite victims and/or their families. Therefore, improving our understanding of venom-induced muscle damage and determining the underlying mechanisms of muscle degeneration/regeneration following snakebites is critical to developing better strategies to tackle this issue. Here, we analysed intramuscular bleeding and thrombosis in muscle injuries induced by two different snake venom toxins (CAMP—Crotalus atrox metalloprotease (a PIII metalloprotease from the venom of this snake) and a three-finger toxin (CTX, a cardiotoxin from the venom of Naja pallida)). Classically, these toxins represent diverse scenarios characterised by persistent muscle damage (CAMP) and successful regeneration (CTX) following acute damage, as normally observed in envenomation by most vipers and some elapid snakes of Asian, Australasian, and African origin, respectively. Our immunohistochemical analysis confirmed that both CAMP and CTX induced extensive muscle destruction on day 5, although the effects of CTX were reversed over time. We identified the presence of fibrinogen and P-selectin exposure inside the damaged muscle sections, suggesting signs of bleeding and the formation of platelet aggregates/microthrombi in tissues, respectively. Intriguingly, CAMP causes integrin shedding but does not affect any blood clotting parameters, whereas CTX significantly extends the clotting time and has no impact on integrin shedding. The rates of fibrinogen clearance and reduction in microthrombi were greater in CTX-treated muscle compared to CAMP-treated muscle. Together, these findings reveal novel aspects of venom-induced muscle damage and highlight the relevance of haemostatic events such as bleeding and thrombosis for muscle regeneration and provide useful mechanistic insights for developing better therapeutic interventions. Full article

(This article belongs to the Special Issue Pre-clinical and Clinical Management of Snakebite Envenomation)

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17 pages, 7408 KB

Open AccessEditor’s ChoiceArticle

A Multi-Layer-Controlled Strategy for Cloning and Expression of Toxin Genes in Escherichia coli

by Jessie Vandierendonck, Yana Girardin, Pieter De Bruyn, Henri De Greve and Remy Loris

Toxins 2023, 15(8), 508; https://doi.org/10.3390/toxins15080508 - 18 Aug 2023

Cited by 6 | Viewed by 6580

Abstract

Molecular cloning and controlled expression remain challenging when the target gene encodes a protein that is toxic to the host. We developed a set of multi-layer control systems to enable cloning of genes encoding proteins known to be highly toxic in Escherichia coli [...] Read more.

Molecular cloning and controlled expression remain challenging when the target gene encodes a protein that is toxic to the host. We developed a set of multi-layer control systems to enable cloning of genes encoding proteins known to be highly toxic in Escherichia coli and other bacteria. The different multi-layer control systems combine a promoter–operator system on a transcriptional level with a riboswitch for translational control. Additionally, replicational control is ensured by using a strain that reduces the plasmid copy number. The use of weaker promoters (such as P_BAD or PfdeA) in combination with the effective theophylline riboswitch is essential for cloning genes that encode notoriously toxic proteins that directly target translation and transcription. Controlled overexpression is possible, allowing the system to be used for evaluating in vivo effects of the toxin. Systems with a stronger promoter can be used for successful overexpression and purification of the desired protein but are limited to toxins that are more moderate and do not interfere with their own production. Full article

(This article belongs to the Section Bacterial Toxins)

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16 pages, 2621 KB

Open AccessEditor’s ChoiceArticle

A Metalloproteinase Cocktail from the Venom of Protobothrops flavoviridis Cleaves Amyloid Beta Peptides at the α-Cleavage Site

by Eugene Futai, Hajime Kawasaki, Shinichi Sato, Khadija Daoudi, Masafumi Hidaka, Taisuke Tomita and Tomohisa Ogawa

Toxins 2023, 15(8), 500; https://doi.org/10.3390/toxins15080500 - 12 Aug 2023

Cited by 5 | Viewed by 5127

Abstract

A disintegrin and metalloproteinase (ADAM) family proteins are a major class of membrane-anchored multidomain proteinases that are responsible for the shedding of cell surface protein ectodomains, including amyloid precursor protein (APP). Human ADAM 9, 10, and 17 proteolyze APPs and produce non-amyloid-genic p3 [...] Read more.

A disintegrin and metalloproteinase (ADAM) family proteins are a major class of membrane-anchored multidomain proteinases that are responsible for the shedding of cell surface protein ectodomains, including amyloid precursor protein (APP). Human ADAM 9, 10, and 17 proteolyze APPs and produce non-amyloid-genic p3 peptides, instead of neurotoxic amyloid-β peptides (Aβs; Aβ40 and Aβ42), which form fibrils and accumulate in the brain of patients with Alzheimer’s disease (AD). The ADAM family is closely related to snake venom metalloproteinases (SVMPs), which are derived from ancestral ADAMs but act as soluble proteinases. To test the therapeutic potential of SVMPs, we purified SVMPs from Protobothrops flavoviridis venom using metal ion affinity and pooled into a cocktail. Thus, 9 out of 11 SVMPs in the P. flavoviridis genome were identified in the cocktail. SVMPs inhibited Aβ secretion when added to human cell culture medium without affecting APP proteolysis. SVMPs degraded synthetic Aβ40 and Aβ42 peptides at the same cleavage site (α-site of APP) as ADAM9, 10, and 17. SVMPs did not degrade Aβ fibrils but interfered with their formation, assessed using thioflavin-T. Thus, SVMPs have therapeutic potential for AD as an Aβ-degrading protease, and the finding adds to the discovery of bioactive peptides from venoms as novel therapeutics. Full article

(This article belongs to the Special Issue Potential Therapeutic Applications of Animal Venoms and Toxins)

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17 pages, 2440 KB

Open AccessEditor’s ChoiceArticle

Characterization of Particle-Size-Based Homogeneity and Mycotoxin Distribution Using Laser Diffraction Particle Size Analysis

by Kai Zhang, Ivy Tran and Steven Tan

Toxins 2023, 15(7), 450; https://doi.org/10.3390/toxins15070450 - 6 Jul 2023

Cited by 9 | Viewed by 5650

Abstract

Sample homogeneity dictates whether analyzing a test portion of an entire sample can provide representative information about incurred mycotoxins. In this study, we evaluated particle-size-distribution-based homogeneity of laboratory mycotoxin samples using laser diffraction particle size analysis and International Organization for Standardization (ISO) Guide [...] Read more.

Sample homogeneity dictates whether analyzing a test portion of an entire sample can provide representative information about incurred mycotoxins. In this study, we evaluated particle-size-distribution-based homogeneity of laboratory mycotoxin samples using laser diffraction particle size analysis and International Organization for Standardization (ISO) Guide 35: 2017. Incurred whole corn, compound feed, peanut butter, and wheat flour (500 g each) were comminuted using wet, cryogenic, or dry milling. We used a sample dividing (riffling) device to obtain representative subsamples (25 g each) and developed a laser diffraction particle size analysis procedure by optimizing key parameters such as the refractive index, absorption, and stirring rate. The homogeneity of the particle size distribution within laboratory subsamples was characterized using the optimized laser diffraction procedure. An assessment of homogeneity was also performed for individual mycotoxins in each incurred matrix sample following the procedure described in ISO Guide 35. The concentrations of the incurred mycotoxins were determined using liquid chromatography–mass spectrometry (LC-MS). Within- and between-subsample variances of incurred aflatoxin B1 in peanut butter; deoxynivalenol in corn, compound feed, and wheat flour; and fumonisins in compound feed corroborated that when the particle size measurements were less than 850 µm, mycotoxins concentrations were consistent across independent test portions, which was confirmed using an analysis of variance (F-test). This study highlights the benefits of laser diffraction particle size analysis and suggests its use as a test procedure to evaluate homogeneity in new sample commodities. Full article

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23 pages, 11155 KB

Open AccessEditor’s ChoiceArticle

Cyanotoxins Increase Cytotoxicity and Promote Nonalcoholic Fatty Liver Disease Progression by Enhancing Cell Steatosis

by Suryakant Niture, Sashi Gadi, Qi Qi, Leslimar Rios-Colon, Sabin Khatiwada, Vandana, Reshan A. Fernando, Keith E. Levine and Deepak Kumar

Toxins 2023, 15(7), 411; https://doi.org/10.3390/toxins15070411 - 25 Jun 2023

Cited by 15 | Viewed by 4199

Abstract

Freshwater prokaryotic cyanobacteria within harmful algal blooms produce cyanotoxins which are considered major pollutants in the aquatic system. Direct exposure to cyanotoxins through inhalation, skin contact, or ingestion of contaminated drinking water can target the liver and may cause hepatotoxicity. In the current [...] Read more.

Freshwater prokaryotic cyanobacteria within harmful algal blooms produce cyanotoxins which are considered major pollutants in the aquatic system. Direct exposure to cyanotoxins through inhalation, skin contact, or ingestion of contaminated drinking water can target the liver and may cause hepatotoxicity. In the current study, we investigated the effect of low concentrations of cyanotoxins on cytotoxicity, inflammation, modulation of unfolded protein response (UPR), steatosis, and fibrosis signaling in human hepatocytes and liver cell models. Exposure to low concentrations of microcystin-LR (MC-LR), microcystin-RR (MC-RR), nodularin (NOD), and cylindrospermopsin (CYN) in human bipotent progenitor cell line HepaRG and hepatocellular carcinoma (HCC) cell lines HepG2 and SK-Hep1 resulted in increased cell toxicity. MC-LR, NOD, and CYN differentially regulated inflammatory signaling, activated UPR signaling and lipogenic gene expression, and induced cellular steatosis and fibrotic signaling in HCC cells. MC-LR, NOD, and CYN also regulated AKT/mTOR signaling and inhibited autophagy. Chronic exposure to MC-LR, NOD, and CYN upregulated the expression of lipogenic and fibrosis biomarkers. Moreover, RNA sequencing (RNA seq) data suggested that exposure of human hepatocytes, HepaRG, and HCC HepG2 cells to MC-LR and CYN modulated expression levels of several genes that regulate non-alcoholic fatty liver disease (NAFLD). Our data suggest that low concentrations of cyanotoxins can cause hepatotoxicity and cell steatosis and promote NAFLD progression. Full article

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31 pages, 7206 KB

Open AccessEditor’s ChoiceArticle

Diversity of Mycotoxins and Other Secondary Metabolites Recovered from Blood Oranges Infected by Colletotrichum, Alternaria, and Penicillium Species

by Ermes Ivan Rovetto, Carlos Luz, Federico La Spada, Giuseppe Meca, Mario Riolo and Santa Olga Cacciola

Toxins 2023, 15(7), 407; https://doi.org/10.3390/toxins15070407 - 21 Jun 2023

Cited by 33 | Viewed by 6070

Abstract

This study identified secondary metabolites produced by Alternaria alternata, Colletotrichum gloeosporioides, and Penicillium digitatum in fruits of two blood orange cultivars before harvest. Analysis was performed by UHPLC–Q-TOF-MS. Three types of fruits were selected, asymptomatic, symptomatic showing necrotic lesions caused by [...] Read more.

This study identified secondary metabolites produced by Alternaria alternata, Colletotrichum gloeosporioides, and Penicillium digitatum in fruits of two blood orange cultivars before harvest. Analysis was performed by UHPLC–Q-TOF-MS. Three types of fruits were selected, asymptomatic, symptomatic showing necrotic lesions caused by hail, and mummified. Extracts from peel and juice were analyzed separately. Penicillium digitatum was the prevalent species recovered from mummified and hail-injured fruits. Among 47 secondary metabolites identified, 16, 18, and 13 were of A. alternata, C. gloeosporioides, and P. digitatum, respectively. Consistently with isolations, indicating the presence of these fungi also in asymptomatic fruits, the metabolic profiles of the peel of hail-injured and asymptomatic fruits did not differ substantially. Major differences were found in the profiles of juice from hail-injured and mummified fruits, such as a significant higher presence of 5,4-dihydroxy-3,7,8-trimethoxy-6C-methylflavone and Atrovenetin, particularly in the juice of mummified fruits of the Tarocco Lempso cultivar. Moreover, the mycotoxins patulin and Rubratoxin B were detected exclusively in mummified fruits. Patulin was detected in both the juice and peel, with a higher relative abundance in the juice, while Rubratoxin B was detected only in the juice. These findings provide basic information for evaluating and preventing the risk of contamination by mycotoxins in the citrus fresh fruit supply chain and juice industry. Full article

(This article belongs to the Section Mycotoxins)

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23 pages, 5583 KB

Open AccessEditor’s ChoiceArticle

The Clostridium botulinum C2 Toxin Subunit C2IIa Delivers Enzymes with Positively Charged N-Termini into the Cytosol of Target Cells

by Sebastian Heber, Joscha Borho, Nicole Stadler, Fanny Wondany, Irina König, Jens Michaelis, Panagiotis Papatheodorou, Holger Barth and Maximilian Fellermann

Toxins 2023, 15(6), 390; https://doi.org/10.3390/toxins15060390 - 9 Jun 2023

Cited by 9 | Viewed by 4374

Abstract

The binary Clostridium (C.) botulinum C2 toxin consists of two non-linked proteins. The proteolytically activated binding/transport subunit C2IIa forms barrel-shaped homoheptamers, which bind to cell surface receptors, mediate endocytosis, and translocate the enzyme subunit C2I into the cytosol of target cells. [...] Read more.

The binary Clostridium (C.) botulinum C2 toxin consists of two non-linked proteins. The proteolytically activated binding/transport subunit C2IIa forms barrel-shaped homoheptamers, which bind to cell surface receptors, mediate endocytosis, and translocate the enzyme subunit C2I into the cytosol of target cells. Here, we investigate whether C2IIa can be harnessed as a transporter for proteins/enzymes fused to polycationic tags, as earlier demonstrated for the related anthrax toxin transport subunit PA₆₃. To test C2IIa-mediated transport in cultured cells, reporter enzymes are generated by fusing different polycationic tags to the N- or C-terminus of other bacterial toxins’ catalytic A subunits. C2IIa as well as PA₆₃ deliver N-terminally polyhistidine-tagged proteins more efficiently compared to C-terminally tagged ones. However, in contrast to PA₆₃, C2IIa does not efficiently deliver polylysine-tagged proteins into the cytosol of target cells. Moreover, untagged enzymes with a native cationic N-terminus are efficiently transported by both C2IIa and PA₆₃. In conclusion, the C2IIa-transporter serves as a transport system for enzymes that harbor positively charged amino acids at their N-terminus. The charge distribution at the N-terminus of cargo proteins and their ability to unfold in the endosome and subsequently refold in the cytosol determine transport feasibility and efficiency. Full article

(This article belongs to the Special Issue Intracellular Transport of Toxins: Insights into Mechanisms of Cytotoxicity and Applications in Cell Biology and Medicine)

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15 pages, 7662 KB

Open AccessFeature PaperEditor’s ChoiceArticle

Domperidone Protects Cells from Intoxication with Clostridioides difficile Toxins by Inhibiting Hsp70-Assisted Membrane Translocation

by Maria Braune-Yan, Jinfang Jia, Mary Wahba, Johannes Schmid, Panagiotis Papatheodorou, Holger Barth and Katharina Ernst

Toxins 2023, 15(6), 384; https://doi.org/10.3390/toxins15060384 - 7 Jun 2023

Cited by 5 | Viewed by 3180

Abstract

Clostridioides difficile infections cause severe symptoms ranging from diarrhea to pseudomembranous colitis due to the secretion of AB-toxins, TcdA and TcdB. Both toxins are taken up into cells through receptor-mediated endocytosis, autoproteolytic processing and translocation of their enzyme domains from acidified endosomes into [...] Read more.

Clostridioides difficile infections cause severe symptoms ranging from diarrhea to pseudomembranous colitis due to the secretion of AB-toxins, TcdA and TcdB. Both toxins are taken up into cells through receptor-mediated endocytosis, autoproteolytic processing and translocation of their enzyme domains from acidified endosomes into the cytosol. The enzyme domains glucosylate small GTPases such as Rac1, thereby inhibiting processes such as actin cytoskeleton regulation. Here, we demonstrate that specific pharmacological inhibition of Hsp70 activity protected cells from TcdB intoxication. In particular, the established inhibitor VER-155008 and the antiemetic drug domperidone, which was found to be an Hsp70 inhibitor, reduced the number of cells with TcdB-induced intoxication morphology in HeLa, Vero and intestinal CaCo-2 cells. These drugs also decreased the intracellular glucosylation of Rac1 by TcdB. Domperidone did not inhibit TcdB binding to cells or enzymatic activity but did prevent membrane translocation of TcdB’s glucosyltransferase domain into the cytosol. Domperidone also protected cells from intoxication with TcdA as well as CDT toxin produced by hypervirulent strains of Clostridioides difficile. Our results reveal Hsp70 requirement as a new aspect of the cellular uptake mechanism of TcdB and identified Hsp70 as a novel drug target for potential therapeutic strategies required to combat severe Clostridioides difficile infections. Full article

(This article belongs to the Special Issue Toxin-Host Interaction of Clostridium Toxins)

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22 pages, 2662 KB

Open AccessFeature PaperEditor’s ChoiceArticle

One Size Fits All—Venomics of the Iberian Adder (Vipera seoanei, Lataste 1878) Reveals Low Levels of Venom Variation across Its Distributional Range

by Ignazio Avella, Maik Damm, Inês Freitas, Wolfgang Wüster, Nahla Lucchini, Óscar Zuazo, Roderich D. Süssmuth and Fernando Martínez-Freiría

Toxins 2023, 15(6), 371; https://doi.org/10.3390/toxins15060371 - 1 Jun 2023

Cited by 18 | Viewed by 6202

Abstract

European vipers (genus Vipera) are medically important snakes displaying considerable venom variation, occurring at different levels in this group. The presence of intraspecific venom variation, however, remains understudied in several Vipera species. Vipera seoanei is a venomous snake endemic to the northern [...] Read more.

European vipers (genus Vipera) are medically important snakes displaying considerable venom variation, occurring at different levels in this group. The presence of intraspecific venom variation, however, remains understudied in several Vipera species. Vipera seoanei is a venomous snake endemic to the northern Iberian Peninsula and south-western France, presenting notable phenotypic variation and inhabiting several diverse habitats across its range. We analysed the venoms of 49 adult specimens of V. seoanei from 20 localities across the species’ Iberian distribution. We used a pool of all individual venoms to generate a V. seoanei venom reference proteome, produced SDS-PAGE profiles of all venom samples, and visualised patterns of variation using NMDS. By applying linear regression, we then assessed presence and nature of venom variation between localities, and investigated the effect of 14 predictors (biological, eco-geographic, genetic) on its occurrence. The venom comprised at least 12 different toxin families, of which five (i.e., PLA₂, svSP, DI, snaclec, svMP) accounted for about 75% of the whole proteome. The comparative analyses of the SDS-PAGE venom profiles showed them to be remarkably similar across the sampled localities, suggesting low geographic variability. The regression analyses suggested significant effects of biological and habitat predictors on the little variation we detected across the analysed V. seoanei venoms. Other factors were also significantly associated with the presence/absence of individual bands in the SDS-PAGE profiles. The low levels of venom variability we detected within V. seoanei might be the result of a recent population expansion, or of processes other than directional positive selection. Full article

(This article belongs to the Section Animal Venoms)

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15 pages, 1860 KB

Open AccessFeature PaperEditor’s ChoiceArticle

Next-Generation Sequencing for Venomics: Application of Multi-Enzymatic Limited Digestion for Inventorying the Snake Venom Arsenal

by Fernanda Gobbi Amorim, Damien Redureau, Thomas Crasset, Lou Freuville, Dominique Baiwir, Gabriel Mazzucchelli, Stefanie K. Menzies, Nicholas R. Casewell and Loïc Quinton

Toxins 2023, 15(6), 357; https://doi.org/10.3390/toxins15060357 - 25 May 2023

Cited by 10 | Viewed by 4586

Abstract

To improve the characterization of snake venom protein profiles, we report the application of a new generation of proteomic methodology to deeply characterize complex protein mixtures. The new approach, combining a synergic multi-enzymatic and a time-limited digestion (MELD), is a versatile and straightforward [...] Read more.

To improve the characterization of snake venom protein profiles, we report the application of a new generation of proteomic methodology to deeply characterize complex protein mixtures. The new approach, combining a synergic multi-enzymatic and a time-limited digestion (MELD), is a versatile and straightforward protocol previously developed by our group. The higher number of overlapping peptides generated during MELD increases the quality of downstream peptide sequencing and of protein identification. In this context, this work aims at applying the MELD strategy to a venomics purpose for the first time, and especially for the characterization of snake venoms. We used four venoms as the test models for this proof of concept: two Elapidae (Dendroaspis polylepis and Naja naja) and two Viperidae (Bitis arietans and Echis ocellatus). Each venom was reduced and alkylated before being submitted to two different protocols: the classical bottom-up proteomics strategy including a digestion step with trypsin only, or MELD, which combines the activities of trypsin, Glu-C and chymotrypsin with a limited digestion approach. The resulting samples were then injected on an M-Class chromatographic system, and hyphenated to a Q-Exactive Mass Spectrometer. Toxins and protein identification were performed by Peaks Studio X+. The results show that MELD considerably improves the number of sequenced (de novo) peptides and identified peptides from protein databases, leading to the unambiguous identification of a greater number of toxins and proteins. For each venom, MELD was successful, not only in terms of the identification of the major toxins (increasing of sequence coverage), but also concerning the less abundant cellular components (identification of new groups of proteins). In light of these results, MELD represents a credible methodology to be applied as the next generation of proteomics approaches dedicated to venomic analysis. It may open new perspectives for the sequencing and inventorying of the venom arsenal and should expand global knowledge about venom composition. Full article

(This article belongs to the Special Issue Omics Approaches to Study Toxins)

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34 pages, 3313 KB

Open AccessEditor’s ChoiceArticle

MicotoXilico: An Interactive Database to Predict Mutagenicity, Genotoxicity, and Carcinogenicity of Mycotoxins

by Josefa Tolosa, Eva Serrano Candelas, José Luis Vallés Pardo, Addel Goya, Salvador Moncho, Rafael Gozalbes and Martina Palomino Schätzlein

Toxins 2023, 15(6), 355; https://doi.org/10.3390/toxins15060355 - 24 May 2023

Cited by 13 | Viewed by 5933

Abstract

Mycotoxins are secondary metabolites produced by certain filamentous fungi. They are common contaminants found in a wide variety of food matrices, thus representing a threat to public health, as they can be carcinogenic, mutagenic, or teratogenic, among other toxic effects. Several hundreds of [...] Read more.

Mycotoxins are secondary metabolites produced by certain filamentous fungi. They are common contaminants found in a wide variety of food matrices, thus representing a threat to public health, as they can be carcinogenic, mutagenic, or teratogenic, among other toxic effects. Several hundreds of mycotoxins have been reported, but only a few of them are regulated, due to the lack of data regarding their toxicity and mechanisms of action. Thus, a more comprehensive evaluation of the toxicity of mycotoxins found in foodstuffs is required. In silico toxicology approaches, such as Quantitative Structure–Activity Relationship (QSAR) models, can be used to rapidly assess chemical hazards by predicting different toxicological endpoints. In this work, for the first time, a comprehensive database containing 4360 mycotoxins classified in 170 categories was constructed. Then, specific robust QSAR models for the prediction of mutagenicity, genotoxicity, and carcinogenicity were generated, showing good accuracy, precision, sensitivity, and specificity. It must be highlighted that the developed QSAR models are compliant with the OECD regulatory criteria, and they can be used for regulatory purposes. Finally, all data were integrated into a web server that allows the exploration of the mycotoxin database and toxicity prediction. In conclusion, the developed tool is a valuable resource for scientists, industry, and regulatory agencies to screen the mutagenicity, genotoxicity, and carcinogenicity of non-regulated mycotoxins. Full article

(This article belongs to the Special Issue Mycotoxins and Fungal Toxins: Current Status and Future Perspectives)

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13 pages, 2421 KB

Open AccessEditor’s ChoiceArticle

Bioactives Overproduction through Operational Strategies in the Ichthyotoxic Microalga Heterosigma akashiwo Culture

by Adrián Macías-de la Rosa, Miguel Ángel González-Cardoso, María del Carmen Cerón-García, Lorenzo López-Rosales, Juan José Gallardo-Rodríguez, Sergio Seoane, Asterio Sánchez-Mirón and Francisco García-Camacho

Toxins 2023, 15(5), 349; https://doi.org/10.3390/toxins15050349 - 20 May 2023

Cited by 7 | Viewed by 3396

Abstract

The red tide-forming microalga Heterosigma akashiwo has been associated with massive events of fish deaths, both wild and cultured. Culture conditions are responsible for the synthesis or accumulation of some metabolites with different interesting bioactivities. H. akashiwo LC269919 strain was grown in a [...] Read more.

The red tide-forming microalga Heterosigma akashiwo has been associated with massive events of fish deaths, both wild and cultured. Culture conditions are responsible for the synthesis or accumulation of some metabolites with different interesting bioactivities. H. akashiwo LC269919 strain was grown in a 10 L bubble column photobioreactor artificially illuminated with multi-coloured LED lights. Growth and production of exopolysaccharides, polyunsaturated fatty acids (PUFAs), and carotenoids were evaluated under different culture modes (batch, fed-batch, semicontinuous, and continuous) at two irradiance levels (300 and 700 µE·s⁻¹·m⁻²). Continuous mode at the dilution rate of 0.2·day⁻¹ and 700 µE·s⁻¹·m⁻² provided the highest production of biomass, PUFAs (132.6 and 2.3 mg·L⁻¹·day⁻¹), and maximum fucoxanthin productivity (0.16 mg·L⁻¹·day⁻¹). The fed-batch mode accumulated exopolysaccharides in a concentration (1.02 g·L⁻¹) 10-fold over the batch mode. An extraction process based on a sequential gradient partition with water and four water-immiscible organic solvents allowed the isolation of bioactive fucoxanthin from methanolic extracts of H. akashiwo. Metabolites present in H. akashiwo, fucoxanthin and polar lipids (i.e., eicosapentaenoic acid (EPA)), or probably such as phytosterol (β-Sitosterol) from other microalgae, were responsible for the antitumor activity obtained. Full article

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

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16 pages, 2106 KB

Open AccessFeature PaperEditor’s ChoiceArticle

A Three-Monoclonal Antibody Combination Potently Neutralizes BoNT/G Toxin in Mice

by Yongfeng Fan, Jianlong Lou, Christina C. Tam, Weihua Wen, Fraser Conrad, Priscila Leal da Silva Alves, Luisa W. Cheng, Consuelo Garcia-Rodriguez, Shauna Farr-Jones and James D. Marks

Toxins 2023, 15(5), 316; https://doi.org/10.3390/toxins15050316 - 30 Apr 2023

Cited by 9 | Viewed by 3691

Abstract

Equine-derived antitoxin (BAT^®) is the only treatment for botulism from botulinum neurotoxin serotype G (BoNT/G). BAT^® is a foreign protein with potentially severe adverse effects and is not renewable. To develop a safe, more potent, and renewable antitoxin, humanized monoclonal [...] Read more.

Equine-derived antitoxin (BAT^®) is the only treatment for botulism from botulinum neurotoxin serotype G (BoNT/G). BAT^® is a foreign protein with potentially severe adverse effects and is not renewable. To develop a safe, more potent, and renewable antitoxin, humanized monoclonal antibodies (mAbs) were generated. Yeast displayed single chain Fv (scFv) libraries were prepared from mice immunized with BoNT/G and BoNT/G domains and screened with BoNT/G using fluorescence-activated cell sorting (FACS). Fourteen scFv-binding BoNT/G were isolated with K_D values ranging from 3.86 nM to 103 nM (median K_D 20.9 nM). Five mAb-binding non-overlapping epitopes were humanized and affinity matured to create antibodies hu6G6.2, hu6G7.2, hu6G9.1, hu6G10, and hu6G11.2, with IgG K_D values ranging from 51 pM to 8 pM. Three IgG combinations completely protected mice challenged with 10,000 LD₅₀s of BoNT/G at a total mAb dose of 6.25 μg per mouse. The mAb combinations have the potential for use in the diagnosis and treatment of botulism due to serotype G and, along with antibody combinations to BoNT/A, B, C, D, E, and F, provide the basis for a fully recombinant heptavalent botulinum antitoxin to replace the legacy equine product. Full article

(This article belongs to the Special Issue Human Antibody Engineering for Prevention and Treatment of Botulism)

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10 pages, 1805 KB

Open AccessEditor’s ChoiceArticle

The Co-Occurrence of T-2 Toxin, Deoxynivalenol, and Fumonisin B1 Activated the Glutathione Redox System in the EU-Limiting Doses in Laying Hens

by Szabina Kulcsár, Benjámin Kövesi, Krisztián Balogh, Erika Zándoki, Zsolt Ancsin, Márta Erdélyi and Miklós Mézes

Toxins 2023, 15(5), 305; https://doi.org/10.3390/toxins15050305 - 23 Apr 2023

Cited by 18 | Viewed by 4115

Abstract

Different mycotoxins in feed lead to combined exposure, increasing adverse effects on animal health. Trichothecene mycotoxins have been associated with inducing oxidative stress, which is neutralized by the glutathione system within the antioxidant defense, depending on the dose and duration of exposure. T-2 [...] Read more.

Different mycotoxins in feed lead to combined exposure, increasing adverse effects on animal health. Trichothecene mycotoxins have been associated with inducing oxidative stress, which is neutralized by the glutathione system within the antioxidant defense, depending on the dose and duration of exposure. T-2 toxin, deoxynivalenol (DON), and fumonisin B1 (FB1) are commonly found in feed commodities simultaneously. In the present study, the intracellular biochemical and gene expression changes were investigated in the case of multi-mycotoxin exposure, focusing on certain elements of the glutathione redox system. In a short-term feeding trial, an in vivo study was performed with low (EU-proposed) doses: T-2/HT-2 toxin: 0.25 mg; DON/2-AcDON/15-AcDON.: 5 mg; FB1: 20 mg/kg feed, and high doses (twice the low dose) in laying hens. The multi-mycotoxin exposure affected the glutathione system; GSH concentration and GPx activity was higher in the liver in the low-dose group on day 1 compared to the control. Furthermore, the gene expression of antioxidant enzymes increased significantly on day 1 in both exposure levels compared to the control. The results suggest that when EU-limiting doses are applied, individual mycotoxins may have a synergistic effect in the induction of oxidative stress. Full article

(This article belongs to the Section Mycotoxins)

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15 pages, 3169 KB

Open AccessEditor’s ChoiceArticle

Immunomodulatory Effects of Cylindrospermopsin in Human T Cells and Monocytes

by Antonio Casas-Rodríguez, Óscar Cebadero-Dominguez, María Puerto, Ana María Cameán and Angeles Jos

Toxins 2023, 15(4), 301; https://doi.org/10.3390/toxins15040301 - 20 Apr 2023

Cited by 18 | Viewed by 3422

Abstract

Cylindrospermopsin (CYN) is a cyanotoxin with an increasing occurrence, and therefore it is important to elucidate its toxicity profile. CYN has been classified as a cytotoxin, although the scientific literature has already revealed that it affects a wide range of organs and systems. [...] Read more.

Cylindrospermopsin (CYN) is a cyanotoxin with an increasing occurrence, and therefore it is important to elucidate its toxicity profile. CYN has been classified as a cytotoxin, although the scientific literature has already revealed that it affects a wide range of organs and systems. However, research on its potential immunotoxicity is still limited. Thus, this study aimed to evaluate the impact of CYN on two human cell lines representative of the immune system: THP-1 (monocytes) and Jurkat (lymphocytes). CYN reduced cell viability, leading to mean effective concentrations (EC50 24 h) of 6.00 ± 1.04 µM and 5.20 ± 1.20 µM for THP-1 and Jurkat cells, respectively, and induced cell death mainly by apoptosis in both experimental models. Moreover, CYN decreased the differentiation of monocytes to macrophages after 48 h of exposure. In addition, an up-regulation of the mRNA expression of different cytokines, such as interleukin (IL) 2, IL-8, tumor necrosis factor-alpha (TNF-α) and interferon-gamma (INF-γ), was also observed mainly after 24 h exposure in both cell lines. However, only an increase in TNF-α in THP-1 supernatants was observed by ELISA. Overall, these results suggest the immunomodulatory activity of CYN in vitro. Therefore, further research is required to evaluate the impact of CYN on the human immune system. Full article

(This article belongs to the Special Issue Toxicology Research on Cyanotoxins)

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22 pages, 3174 KB

Open AccessFeature PaperEditor’s ChoiceArticle

Application of an Extracellular Matrix-Mimicking Fluorescent Polymer for the Detection of Proteolytic Venom Toxins

by Eric Wachtel, Matyas A. Bittenbinder, Bas van de Velde, Julien Slagboom, Axel de Monts de Savasse, Luis L. Alonso, Nicholas R. Casewell, Freek J. Vonk and Jeroen Kool

Toxins 2023, 15(4), 294; https://doi.org/10.3390/toxins15040294 - 18 Apr 2023

Cited by 3 | Viewed by 3718

Abstract

The cytotoxicity caused by snake venoms is a serious medical problem that greatly contributes to the morbidity observed in snakebite patients. The cytotoxic components found in snake venoms belong to a variety of toxin classes and may cause cytotoxic effects by targeting a [...] Read more.

The cytotoxicity caused by snake venoms is a serious medical problem that greatly contributes to the morbidity observed in snakebite patients. The cytotoxic components found in snake venoms belong to a variety of toxin classes and may cause cytotoxic effects by targeting a range of molecular structures, including cellular membranes, the extracellular matrix (ECM) and the cytoskeleton. Here, we present a high-throughput assay (384-well plate) that monitors ECM degradation by snake venom toxins via the application of fluorescent versions of model ECM substrates, specifically gelatin and collagen type I. Both crude venoms and fractionated toxins of a selection of medically relevant viperid and elapid species, separated via size-exclusion chromatography, were studied using the self-quenching, fluorescently labelled ECM–polymer substrates. The viperid venoms showed significantly higher proteolytic degradation when compared to elapid venoms, although the venoms with higher snake venom metalloproteinase content did not necessarily exhibit stronger substrate degradation than those with a lower one. Gelatin was generally more readily cleaved than collagen type I. In the viperid venoms, which were subjected to fractionation by SEC, two (B. jararaca and C. rhodostoma, respectively) or three (E. ocellatus) active proteases were identified. Therefore, the assay allows the study of proteolytic activity towards the ECM in vitro for crude and fractionated venoms. Full article

(This article belongs to the Special Issue Animal Venoms: Proteomics, Biochemical Activities and Application)

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12 pages, 1938 KB

Open AccessEditor’s ChoiceArticle

Quantitative and Qualitative Pain Evaluation in Response to OnabotulinumtoxinA for Chronic Migraine: An Observational Real-Life Study

by Claudia Altamura, Nicoletta Brunelli, Giovanna Viticchi, Sergio Salvemini, Gianluca Cecchi, Marilena Marcosano, Luisa Fofi, Mauro Silvestrini and Fabrizio Vernieri

Toxins 2023, 15(4), 284; https://doi.org/10.3390/toxins15040284 - 15 Apr 2023

Cited by 7 | Viewed by 3470

Abstract

(1) Background: Randomized controlled trials and real-life studies demonstrated the efficacy of OnabotulinumtoxinA (OBT-A) for CM prevention. However, no studies specifically addressed its effect on pain’s quantitative intensity and qualitative characteristics. (2) Methods: This is an ambispective study: a post-hoc retrospective analysis of [...] Read more.

(1) Background: Randomized controlled trials and real-life studies demonstrated the efficacy of OnabotulinumtoxinA (OBT-A) for CM prevention. However, no studies specifically addressed its effect on pain’s quantitative intensity and qualitative characteristics. (2) Methods: This is an ambispective study: a post-hoc retrospective analysis of real-life prospectively collected data from two Italian headache centers on CM patients treated with OBT-A over one year (i.e., Cy1-4). The primary endpoint was the changes in pain intensity (Numeric Rating Scale, NRS; the Present Pain Intensity (PPI) scale, the 6-point Behavioral Rating Scale (BRS-6)) and quality scale (the short-form McGill Pain Questionnaire (SF-MPQ)) scores. We also assessed the relationship between changes in intensity and quality of pain and disability scale (MIDAS; HIT-6) scores, monthly headache days (MHDs), and monthly acute medication intake (MAMI) (3) Results: We retrieved 152 cases (51.5 years SD 11.3, 80.3% females). From baseline to Cy-4, MHDs, MAMI, NRS, PPI, and BRS-6 scores decreased (consistently p < 0.001). Only the throbbing (p = 0.004), splitting (p = 0.018), and sickening (p = 0.017) qualities of pain collected in the SF-MPQ were reduced. Score variations in MIDAS related to those in PPI scales (p = 0.035), in the BRS-6 (p = 0.001), and in the NRS (p = 0.003). Similarly, HIT-6 score changes related to PPI score modifications (p = 0.027), in BRS-6 (p = 0.001) and NRS (p = 0.006). Conversely, MAMI variation was not associated with qualitative or quantitative pain score modifications except BRS-6 (p = 0.018). (4) Conclusions: Our study shows that OBT-A alleviates migraine by reducing its impact on multiple aspects, such as frequency, disability, and pain intensity. The beneficial effect on pain intensity seems specific to pain characteristics related to C-fiber transmission and is associated with a reduction in migraine-related disability. Full article

(This article belongs to the Special Issue Botulinum Toxin and Migraine: Goals and Perspectives)

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12 pages, 329 KB

Open AccessEditor’s ChoiceArticle

Proton-Pump Inhibitors and Serum Concentrations of Uremic Toxins in Patients with Chronic Kidney Disease

by Carolla El Chamieh, Islam Amine Larabi, Solène M. Laville, Christian Jacquelinet, Christian Combe, Denis Fouque, Maurice Laville, Luc Frimat, Roberto Pecoits-Filho, Céline Lange, Bénédicte Stengel, Natalia Alencar De Pinho, Jean-Claude Alvarez, Ziad A. Massy and Sophie Liabeuf

Toxins 2023, 15(4), 276; https://doi.org/10.3390/toxins15040276 - 7 Apr 2023

Cited by 5 | Viewed by 5064

Abstract

Use of proton-pump inhibitors (PPIs) is common in patients with chronic kidney disease (CKD). PPIs and many uremic toxins (UTs) are eliminated by the kidney’s tubular organic anion transporter system. In a cross-sectional study, we sought to evaluate the association between PPI prescription [...] Read more.

Use of proton-pump inhibitors (PPIs) is common in patients with chronic kidney disease (CKD). PPIs and many uremic toxins (UTs) are eliminated by the kidney’s tubular organic anion transporter system. In a cross-sectional study, we sought to evaluate the association between PPI prescription and serum concentrations of various UTs. We studied a randomly selected sub-group of participants in the CKD-REIN cohort (adult patients with a confirmed diagnosis of CKD and estimated glomerular filtration rate (eGFR) < 60 mL/min/1.73 m²) with available frozen samples collected at baseline. PPI prescription was recorded at baseline. Serum concentrations of 10 UTs were measured using a validated liquid chromatography tandem mass spectrometry technique. Multiple linear regression was performed, with the log UT concentration as the dependent variable. Of the 680 included patients (median age: 68 years; median eGFR: 32 mL/min/1.73 m²), 31% had PPI prescriptions at baseline. Patients using PPIs had higher levels of certain UTs in comparison to other patients, including total and free indoxyl sulfate (IS), total and free p-cresylsulfate, total and free p-cresylglucuronide (PCG), phenylacetylglutamine (PAG), free kynurenine, and free hippuric acid. After adjustment for baseline co-morbidities, number of co-prescribed drugs, and laboratory data, including eGFR, associations between PPI prescription and elevated serum concentrations of free and total IS, free and total PCG, and PAG remained significant. Our results indicate that PPI prescription is independently associated with serum UT retention. These findings are interesting to better understand the factors that may modulate serum UT concentration in CKD patients, however, they will need to be confirmed by longitudinal studies. Full article

(This article belongs to the Section Uremic Toxins)

17 pages, 2908 KB

Open AccessEditor’s ChoiceArticle

Proteomics of Vespa velutina nigrithorax Venom Sac Queens and Workers: A Quantitative SWATH-MS Analysis

by Manuela Alonso-Sampedro, Xesús Feás, Susana Belén Bravo, María Pilar Chantada-Vázquez and Carmen Vidal

Toxins 2023, 15(4), 266; https://doi.org/10.3390/toxins15040266 - 3 Apr 2023

Cited by 6 | Viewed by 7837

Abstract

Health risks caused by stings from Vespa velutina nigrithorax (VV), also known as the yellow-legged Asian hornet, have become a public concern, but little is known about its venom composition. This study presents the proteome profile of the VV’s venom sac (VS) based on [...] Read more.

Health risks caused by stings from Vespa velutina nigrithorax (VV), also known as the yellow-legged Asian hornet, have become a public concern, but little is known about its venom composition. This study presents the proteome profile of the VV’s venom sac (VS) based on Sequential Window Acquisition of all Theoretical Mass Spectra (SWATH-MS). The study also performed proteomic quantitative analysis and examined the biological pathways and molecular functions of the proteins in the VS of VV gynes (i.e., future queens [SQ]) and workers [SW]. The total protein content per VS was significantly higher in the SW than in the SQ (274 ± 54 µg/sac vs. 175 ± 22 µg/sac; p = 0.02). We quantified a total of 228 proteins in the VS, belonging to 7 different classes: Insecta (n = 191); Amphibia and Reptilia (n = 20); Bacilli, γ-Proteobacteria and Pisoniviricetes (n = 12); and Arachnida (n = 5). Among the 228 identified proteins, 66 showed significant differential expression between SQ and SW. The potential allergens hyaluronidase A, venom antigen 5 and phospholipase A1 were significantly downregulated in the SQ venom. Full article

(This article belongs to the Special Issue Advances in Venom Immunology and Allergy)

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