Toxins

14 pages, 2748 KiB

Open AccessArticle

Identification of New Angiotensin-Converting Enzyme Inhibitory Peptides Isolated from the Hydrolysate of the Venom of Nemopilema nomurai Jellyfish

by Ramachandran Loganathan Mohan Prakash, Deva Asirvatham Ravi, Du Hyeon Hwang, Changkeun Kang and Euikyung Kim

Toxins 2024, 16(9), 410; https://doi.org/10.3390/toxins16090410 - 20 Sep 2024

Viewed by 299

Abstract

Recently, jellyfish venom has gained attention as a promising reservoir of pharmacologically active compounds, with potential applications in new drug development. In this investigation, novel peptides, isolated from the hydrolysates of Nemopilema nomurai jellyfish venom (NnV), demonstrate potent inhibitory activities against angiotensin-converting enzyme [...] Read more.

Recently, jellyfish venom has gained attention as a promising reservoir of pharmacologically active compounds, with potential applications in new drug development. In this investigation, novel peptides, isolated from the hydrolysates of Nemopilema nomurai jellyfish venom (NnV), demonstrate potent inhibitory activities against angiotensin-converting enzyme (ACE). Proteolytic enzymes—specifically, papain and protamex—were utilized for the hydrolysis under optimized enzymatic conditions, determined by assessing the degree of hydrolysis through the ninhydrin test. Comparative analyses revealed that papain treatment exhibited a notably higher degree of NnV hydrolysis compared to protamex treatment. ACE inhibitory activity was quantified using ACE kit-WST, indicating a substantial inhibitory effect of 76.31% for the papain-digested NnV crude hydrolysate, which was validated by captopril as a positive control. The separation of the NnV-hydrolysate using DEAE sepharose weak-anion-exchange chromatography revealed nine peaks under a 0–1 M NaCl stepwise gradient, with peak no. 3 displaying the highest ACE inhibition of 96%. The further purification of peak no. 3 through ODS-C18 column reverse-phase high-performance liquid chromatography resulted in five sub-peaks (3.1, 3.2, 3.3, 3.4, and 3.5), among which 3.2 exhibited the most significant inhibitory activity of 95.74%. The subsequent analysis of the active peak (3.2) using MALDI–TOF/MS identified two peptides with distinct molecular weights of 896.48 and 1227.651. The peptide sequence determined by MS/MS analysis revealed them as IVGRPLANG and IGDEPRHQYL. The docking studies of the two ACE-inhibitory peptides for ACE molecule demonstrated a binding affinity of −51.4 ± 2.5 and −62.3 ± 3.3 using the HADDOCK scoring function. Full article

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

► Show Figures

Figure 1

13 pages, 2192 KiB

Open AccessArticle

The Role of the N-Terminal Domain of Thrombomodulin and the Potential of Recombinant Human Thrombomodulin as a Therapeutic Intervention for Shiga Toxin-Induced Hemolytic-Uremic Syndrome

by Sarah Kröller, Jana Schober, Nadine Krieg, Sophie Dennhardt, Wiebke Pirschel, Michael Kiehntopf, Edward M. Conway and Sina M. Coldewey

Toxins 2024, 16(9), 409; https://doi.org/10.3390/toxins16090409 - 20 Sep 2024

Viewed by 405

Abstract

Hemolytic-uremic syndrome (HUS) is a rare complication of an infection with Shiga toxin (Stx)-producing Escherichia coli (STEC-HUS), characterized by severe acute kidney injury, thrombocytopenia and microangiopathic hemolytic anemia, and specific therapy is still lacking. Thrombomodulin (TM) is a multi-domain transmembrane endothelial cell protein [...] Read more.

Hemolytic-uremic syndrome (HUS) is a rare complication of an infection with Shiga toxin (Stx)-producing Escherichia coli (STEC-HUS), characterized by severe acute kidney injury, thrombocytopenia and microangiopathic hemolytic anemia, and specific therapy is still lacking. Thrombomodulin (TM) is a multi-domain transmembrane endothelial cell protein and its N-terminal domain has been implicated in the pathophysiology of some cases of HUS. Indeed, the administration of recombinant human TM (rhTM) may have efficacy in HUS. We used a Stx-based murine model of HUS to characterize the role of the N-terminal domain of TM. We show that mice lacking that domain (TMLed (−/−)) are more sensitive to Stx, with enhanced HUS progression seen at 4 days and increased mortality at 7 days post-HUS induction. In spite of these changes, renal function was less affected in surviving Stx-challenged TMLed (−/−) mice compared to their wild-type counterparts TMLed (+/+) at 7 days. Contrary to few clinical case reports from Japan, the administration of rhTM (0.06 mg/kg) to wild-type mice (C57BL/6J) with HUS did not protect against disease progression. This overall promising, but also contradictory body of evidence, requires further systematic preclinical and clinical investigations to clarify the role of TM in HUS as a potential therapeutic strategy. Full article

► Show Figures

Figure 1

13 pages, 3348 KiB

Open AccessArticle

The Emerging Fusarium graminearum NA3 Population Produces High Levels of Mycotoxins in Wheat and Barley

by Nicholas A. Rhoades, Susan P. McCormick, Martha M. Vaughan and Guixia Hao

Toxins 2024, 16(9), 408; https://doi.org/10.3390/toxins16090408 - 20 Sep 2024

Viewed by 637

Abstract

Fusarium graminearum (Fg) is the primary causal agent of Fusarium head blight (FHB) in wheat, barley, and other small grains in North America and worldwide. FHB results in yield reduction and contaminates grain with mycotoxins that pose threats to human and [...] Read more.

Fusarium graminearum (Fg) is the primary causal agent of Fusarium head blight (FHB) in wheat, barley, and other small grains in North America and worldwide. FHB results in yield reduction and contaminates grain with mycotoxins that pose threats to human and livestock health. Three genetically distinct North American (NA) populations of Fg have been characterized, which are generally associated with differences in their predominant trichothecene chemotype: NA1/15-acetyl-deoxynivalenol (15-ADON), NA2/3-acetyl-deoxynivalenol (3-ADON), and NA3/3α-acetoxy, 7,15-dihydroxy-12,13-epoxytrichothec-9-ene (NX-2). Recent studies found that the NA3 population had significantly less spread on point-inoculated wheat spikes than the NA1 and NA2 populations, and NX toxins are important for Fg spread and initial infection in wheat. In this follow-up study, to compare the effect of the three populations on initial infection and mycotoxin production on different hosts, we dip-inoculated spikes of the moderately resistant wheat cultivar Alsen and the susceptible barley cultivar Voyager using five strains from each population to evaluate disease, trichothecene mycotoxin accumulation, and trichothecene production per unit of fungal biomass. In dip-inoculated wheat spikes, the NA3 population produced significantly more trichothecene per unit of fungal biomass and accumulated higher levels of trichothecene per plant biomass than the NA1 and NA2 populations, regardless of the disease levels caused by the three populations. In contrast to its critical role during wheat infection, NX toxins had no effect on barley infection. In dip-inoculated barley, the NA1 population was more infectious and caused more severe FHB symptoms than the NA2 and NA3 populations; however, the NA3 population produced significantly higher toxin per unit of fungal biomass in infected barley tissues than the NA1 population. This study provides critical information on the emerging NA3 population, which produces high levels of NX toxin and poses a potential food safety concern. Full article

(This article belongs to the Section Mycotoxins)

► Show Figures

Figure 1

17 pages, 20060 KiB

Open AccessArticle

Rab4b Promotes Cytolethal Distending Toxin from Glaesserella parasuis-Induced Cytotoxicity in PK-15 Cells

by Yiwen Zhang, Zhen Yang, Ke Dai, Bangdi Hu, Shiyu Xu, Yu Wang, Li Lei, Senyan Du, Qin Zhao, Xiaobo Huang, Rui Wu, Qigui Yan, Yiping Wang, Sanjie Cao and Yiping Wen

Toxins 2024, 16(9), 407; https://doi.org/10.3390/toxins16090407 - 19 Sep 2024

Viewed by 313

Abstract

Glaesserella parasuis cytolethal distending toxin (GpCDT) can induce cell cycle arrest and apoptosis. Our laboratory’s previous work demonstrated that GTPase 4b (Rab4b) is a key host protein implicated in GpCDT-induced cytotoxicity. This study investigated the probable involvement of Rab4b in [...] Read more.

Glaesserella parasuis cytolethal distending toxin (GpCDT) can induce cell cycle arrest and apoptosis. Our laboratory’s previous work demonstrated that GTPase 4b (Rab4b) is a key host protein implicated in GpCDT-induced cytotoxicity. This study investigated the probable involvement of Rab4b in the process. Our study used CRISPR/Cas9 technology to create a Rab4b-knockout cell line. The results showed greater resistance to GpCDT-induced cell cytotoxicity. In contrast, forced Rab4b overexpression increased GpCDT-induced cytotoxicity. Further immunoprecipitation study reveals that GpCDT may bind with Rab4b. In PK-15 cells, GpCDT is transported to the early endosomes and late endosomes, while after knocking out Rab4b, GpCDT cannot be transported to the early endosome via vesicles. Rab4b appears essential for GpCDT-induced cytotoxicity in PK-15 cells. Full article

(This article belongs to the Section Bacterial Toxins)

► Show Figures

Figure 1

19 pages, 6560 KiB

Open AccessReview

Host Tropism and Structural Biology of ABC Toxin Complexes

by Cole L. Martin, John H. Hill and Stephen G. Aller

Toxins 2024, 16(9), 406; https://doi.org/10.3390/toxins16090406 - 19 Sep 2024

Viewed by 378

Abstract

ABC toxin complexes are a class of protein toxin translocases comprised of a multimeric assembly of protein subunits. Each subunit displays a unique composition, contributing to the formation of a syringe-like nano-machine with natural cargo carrying, targeting, and translocation capabilities. Many of these [...] Read more.

ABC toxin complexes are a class of protein toxin translocases comprised of a multimeric assembly of protein subunits. Each subunit displays a unique composition, contributing to the formation of a syringe-like nano-machine with natural cargo carrying, targeting, and translocation capabilities. Many of these toxins are insecticidal, drawing increasing interest in agriculture for use as biological pesticides. The A subunit (TcA) is the largest subunit of the complex and contains domains associated with membrane permeation and targeting. The B and C subunits, TcB and TcC, respectively, package into a cocoon-like structure that contains a toxic peptide and are coupled to TcA to form a continuous channel upon final assembly. In this review, we outline the current understanding and gaps in the knowledge pertaining to ABC toxins, highlighting seven published structures of TcAs and how these structures have led to a better understanding of the mechanism of host tropism and toxin translocation. We also highlight similarities and differences between homologues that contribute to variations in host specificity and conformational change. Lastly, we review the biotechnological potential of ABC toxins as both pesticides and cargo-carrying shuttles that enable the transport of peptides into cells. Full article

(This article belongs to the Section Bacterial Toxins)

► Show Figures

Figure 1

20 pages, 3697 KiB

Open AccessArticle

Isolation and Pharmacological Characterisation of Pre-Synaptic Neurotoxins from Thai and Javanese Russell’s Viper (Daboia siamensis) Venoms

by Mimi Lay and Wayne C. Hodgson

Toxins 2024, 16(9), 405; https://doi.org/10.3390/toxins16090405 - 19 Sep 2024

Viewed by 316

Abstract

The widespread geographical distribution of Russell’s vipers (Daboia spp.) is associated with marked variations in the clinical outcomes of envenoming by species from different countries. This is likely to be due to differences in the quantity and potency of key toxins and, [...] Read more.

The widespread geographical distribution of Russell’s vipers (Daboia spp.) is associated with marked variations in the clinical outcomes of envenoming by species from different countries. This is likely to be due to differences in the quantity and potency of key toxins and, potentially, the presence or absence of some toxins in venoms across the geographical spectrum. In this study, we aimed to isolate and pharmacologically characterise the major neurotoxic components of D. siamensis venoms from Thailand and Java (Indonesia) and explore the efficacy of antivenom and a PLA₂ inhibitor, Varespladib, against the neuromuscular activity. These data will provide insights into the link between venom components and likely clinical outcomes, as well as potential treatment strategies. Venoms were fractionated using RP-HPLC and the in vitro activity of isolated toxins assessed using the chick biventer cervicis nerve-muscle preparation. Two major PLA₂ fractions (i.e., fractions 8 and 10) were isolated from each venom. Fraction 8 from both venoms produced pre-synaptic neurotoxicity and myotoxicity, whereas fraction 10 from both venoms was weakly neurotoxic. The removal of the two fractions from each venom abolished the in vitro neurotoxicity, and partially abolished myotoxicity, of the whole venom. A combination of the two fractions from each venom produced neurotoxic activity that was equivalent to the respective whole venom (10 µg/mL), but the myotoxic effects were not additive. The in vitro neurotoxicity of fraction 8 (100 nM) from each venom was prevented by the pre-administration of Thai Russell’s viper monovalent antivenom (2× recommended concentration) or preincubation with Varespladib (100 nM). Additionally, the neurotoxicity produced by a combination of the two fractions was partially reversed by the addition of Varespladib (100–300 nM) 60 min after the fractions. The present study demonstrates that the in vitro skeletal muscle effects of Thai and Javanese D. siamensis venoms are primarily due to key PLA₂ toxins in each venom. Full article

(This article belongs to the Special Issue Snake Venom: Toxicology and Associated Countermeasures)

► Show Figures

Figure 1

16 pages, 2284 KiB

Open AccessArticle

Enzymatic Hydrolysis of Resorcylic Acid Lactones by an Aeromicrobium sp.

by Shawn J. Hoogstra, Kyle N. Hendricks, David R. McMullin, Justin B. Renaud, Juhi Bora, Mark W. Sumarah and Christopher P. Garnham

Toxins 2024, 16(9), 404; https://doi.org/10.3390/toxins16090404 - 19 Sep 2024

Viewed by 321

Abstract

Zearalenone and radicicol are resorcylic acid lactones produced by numerous plant pathogenic fungi. Zearalenone is a non-steroidal estrogen mimic that can cause serious reproductive issues in livestock that consume contaminated feed. Radicicol is a potent inhibitor of the molecular chaperone Hsp90, which, in [...] Read more.

Zearalenone and radicicol are resorcylic acid lactones produced by numerous plant pathogenic fungi. Zearalenone is a non-steroidal estrogen mimic that can cause serious reproductive issues in livestock that consume contaminated feed. Radicicol is a potent inhibitor of the molecular chaperone Hsp90, which, in plants, has an important role in coordinating the host’s immune response during infection. Here, we describe the identification and characterization of a soil-borne strain of the Gram-positive bacterium Aeromicrobium sp. capable of hydrolyzing the macrolide ring of resorcylic acid lactones, including zearalenone and radicicol. Proteomic analysis of biochemically enriched fractions from the isolated and cultured bacterium identified an α/β-hydrolase responsible for this activity. A recombinantly expressed and purified form of the hydrolase (termed RALH) was active against both zearalenone and radicicol. Interpretation of high-resolution mass spectrometry and NMR data confirmed the structures of the enzymatic products as the previously reported non-toxic metabolite hydrolyzed zearalenone and hydrolyzed radicicol. Hydrolyzed radicicol was demonstrated to no longer inhibit the ATPase activity of the Saccharomyces cerevisiae Hsp90 homolog in vitro. Enzymatic degradation of resorcylic acid lactones will enable insight into their biological functions. Full article

► Show Figures

Figure 1

17 pages, 3157 KiB

Open AccessArticle

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

Viewed by 393

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)

► Show Figures

Figure 1

21 pages, 4548 KiB

Open AccessArticle

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

Viewed by 383

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)

► Show Figures

Figure 1

15 pages, 7210 KiB

Open AccessArticle

Astaxanthin Alleviates Hepatic Lipid Metabolic Dysregulation Induced by Microcystin-LR

by Qinmei Tan, Hanyu Chu, Jia Wei, Sisi Yan, Xiaoya Sun, Jiangping Wang, Lemei Zhu and Fei Yang

Toxins 2024, 16(9), 401; https://doi.org/10.3390/toxins16090401 - 18 Sep 2024

Viewed by 373

Abstract

Microcystin-LR (MC-LR), frequently generated by cyanobacteria, has been demonstrated to raise the likelihood of liver disease. Few previous studies have explored the potential antagonist against MC-LR. Astaxanthin (ASX) has been shown to possess various beneficial effects in regulating lipid metabolism in the liver. [...] Read more.

Microcystin-LR (MC-LR), frequently generated by cyanobacteria, has been demonstrated to raise the likelihood of liver disease. Few previous studies have explored the potential antagonist against MC-LR. Astaxanthin (ASX) has been shown to possess various beneficial effects in regulating lipid metabolism in the liver. However, whether ASX could alleviate MC-LR-induced hepatic lipid metabolic dysregulation is as yet unclear. In this work, the important roles and mechanisms of ASX in countering MC-LR-induced liver damage and lipid metabolic dysregulation were explored for the first time. The findings revealed that ASX not only prevented weight loss but also enhanced liver health after MC-LR exposure. Moreover, ASX effectively decreased triglyceride, total cholesterol, aspartate transaminase, and alanine aminotransferase contents in mice that were elevated by MC-LR. Histological observation showed that ASX significantly alleviated lipid accumulation and inflammation induced by MC-LR. Mechanically, ASX could significantly diminish the expression of genes responsible for lipid generation (Srebp-1c, Fasn, Cd36, Scd1, Dgat1, and Pparg), which probably reduced lipid accumulation induced by MC-LR. Analogously, MC-LR increased intracellular lipid deposition in THLE-3 cells, while ASX decreased these symptoms by down-regulating the expression of key genes in the lipid synthesis pathway. Our results implied that ASX played a crucial part in lipid synthesis and effectively alleviated MC-LR-induced lipid metabolism dysregulation. ASX might be developed as a novel protectant against hepatic impairment and lipid metabolic dysregulation associated with MC-LR. This study offers new insights for further management of MC-LR-related metabolic diseases. Full article

► Show Figures

Graphical abstract

19 pages, 6017 KiB

Open AccessArticle

The Contrasting Effects of Bothrops lanceolatus and Bothrops atrox Venom on Procoagulant Activity and Thrombus Stability under Blood Flow Conditions

by Fatima Radouani, Prisca Jalta, Caroline Rapon, Chloe Lezin, Chelsea Branford, Jonathan Florentin, Jose Maria Gutierrez, Dabor Resiere, Remi Neviere and Olivier Pierre-Louis

Toxins 2024, 16(9), 400; https://doi.org/10.3390/toxins16090400 - 18 Sep 2024

Viewed by 358

Abstract

Background: Consumption coagulopathy and hemorrhagic syndrome are the typical features of Bothrops sp. snake envenoming. In contrast, B. lanceolatus envenoming can induce thrombotic complications. Our aim was to test whether crude B. lanceolatus and B. atrox venoms would display procoagulant activity and induce [...] Read more.

Background: Consumption coagulopathy and hemorrhagic syndrome are the typical features of Bothrops sp. snake envenoming. In contrast, B. lanceolatus envenoming can induce thrombotic complications. Our aim was to test whether crude B. lanceolatus and B. atrox venoms would display procoagulant activity and induce thrombus formation under flow conditions. Methods and Principal Findings: Fibrin formation in human plasma was observed for B. lanceolatus venom at 250–1000 ng/mL concentrations, which also induced clot formation in purified human fibrinogen, indicating thrombin-like activity. The degradation of fibrinogen confirmed the fibrinogenolytic activity of B. lanceolatus venom. B. lanceolatus venom displayed consistent thrombin-like and kallikrein-like activity increases in plasma conditions. The well-known procoagulant B. atrox venom activated plasmatic coagulation factors in vitro and induced firm thrombus formation under high shear rate conditions. In contrast, B. lanceolatus venom induced the formation of fragile thrombi that could not resist shear stress. Conclusions: Our results suggest that crude B. lanceolatus venom displays amidolytic activity and can activate the coagulation cascade, leading to prothrombin activation. B. lanceolatus venom induces the formation of an unstable thrombus under flow conditions, which can be prevented by the specific monovalent antivenom Bothrofav^®. Full article

(This article belongs to the Section Animal Venoms)

► Show Figures

Figure 1

14 pages, 2179 KiB

Open AccessArticle

Transcriptomic Analysis Reveals Diverse Expression of Scorpion Toxin Genes in Mesobuthus martensii

by Zhongxian Yang, Haiquan Wang, Yan Zhao, Jianyu Huang, Chao Zhang and Zhiyong Di

Toxins 2024, 16(9), 399; https://doi.org/10.3390/toxins16090399 - 18 Sep 2024

Viewed by 347

Abstract

Scorpions, an ancient group of venomous invertebrates, have existed for over 430 million years. Their toxins, important for predation and defense, exhibit a variety of biological and pharmacological activities. Research on scorpion toxins has spanned decades. Notably, the toxin genes of Mesobuthus martensii [...] Read more.

Scorpions, an ancient group of venomous invertebrates, have existed for over 430 million years. Their toxins, important for predation and defense, exhibit a variety of biological and pharmacological activities. Research on scorpion toxins has spanned decades. Notably, the toxin genes of Mesobuthus martensii (Scorpiones: Buthidae), a well-known Chinese herbal medicine, have been described at genomic and proteomic levels. However, previous studies primarily focused on the toxin genes expressed in the venom glands, overlooking their expression in multiple tissues. This study analyzed transcriptomes from 14 tissues of M. martensii. Gene annotation revealed 83 toxin and toxin-like genes, including those affecting sodium, potassium, calcium, and chloride ion channels. Approximately 70% of toxin genes were highly expressed in the vesicle; additionally, some exhibited low or no expression in the vesicle while showing high expression in other tissues. Beyond the vesicle, high expression levels of toxin genes were observed in metasoma segments II-V, blood, lateral eyes, chelicerae, legs, pedipalp chelae, femurs, and patellae. This expression pattern suggests that toxin genes are recruited from multiple tissues and may help prevent intraspecific harm during courtship and competition for prey. These findings inspire further research into the evolutionary recruitment process of scorpion toxins. Full article

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

► Show Figures

Figure 1

15 pages, 2308 KiB

Open AccessArticle

Thermoregulation Effects of Phoneutria nigriventer Isolated Toxins in Rats

by Carla Bogri Butkeraitis, Monica Viviana Abreu Falla and Ivo Lebrun

Toxins 2024, 16(9), 398; https://doi.org/10.3390/toxins16090398 - 18 Sep 2024

Viewed by 435

Abstract

Body temperature is primarily regulated by the hypothalamus, ensuring proper metabolic function. Envenomation by Phoneutria nigriventer can cause symptoms such as hypothermia, hyperthermia, sweating, and shivering, all related to thermoregulation. This study aims to analyze and identify components of the venom that affect [...] Read more.

Body temperature is primarily regulated by the hypothalamus, ensuring proper metabolic function. Envenomation by Phoneutria nigriventer can cause symptoms such as hypothermia, hyperthermia, sweating, and shivering, all related to thermoregulation. This study aims to analyze and identify components of the venom that affect thermoregulation and to evaluate possible mechanisms. Rats were used for thermoregulation analysis, venom fractionation by gel filtration and reverse-phase chromatography (C18), and sequencing by Edman degradation. The venom exhibited hypothermic effects in rats, while its fractions demonstrated both hypothermic (pool II) and hyperthermic (pool III) effects. Further separations of the pools with C18 identified specific peaks responsible for these effects. However, as the peaks were further purified, their effects became less significant. Tests on U87 human glioblastoma cells showed no toxicity. Sequencing of the most active peaks revealed masses similar to those of the Tachykinin and Ctenotoxin families, both known to act on the nervous system. The study concludes that molecules derived from venom can act synergistically or antagonistically. Additionally, toxins that affect thermoregulation are poorly studied and require further characterization. These toxins could potentially serve as sources for the development of new thermoregulatory drugs. Full article

(This article belongs to the Special Issue New Insights into Structure and Function of Arthropod Toxins and Other Toxins: Tribute to Professor Hervé Rochat)

► Show Figures

Graphical abstract

13 pages, 1369 KiB

Open AccessArticle

Global Perspectives on Mycotoxin Reference Materials (Part I): Insights from Multi-Supplier Comparison Study Including Aflatoxin B1, Deoxynivalenol and Zearalenone

by David Steiner, Tibor Bartók, Michael Sulyok, András Szekeres, Mónika Varga, Levente Horváth and Helmut Rost

Toxins 2024, 16(9), 397; https://doi.org/10.3390/toxins16090397 - 17 Sep 2024

Viewed by 492

Abstract

We conducted a comprehensive examination of liquid mycotoxin reference standards. A total of 30 different standards were tested, each containing 10 samples of three distinct substances: Aflatoxin B1, Deoxynivalenol, and Zearalenone. The standards were sourced from 10 different global market leading manufacturers. To [...] Read more.

We conducted a comprehensive examination of liquid mycotoxin reference standards. A total of 30 different standards were tested, each containing 10 samples of three distinct substances: Aflatoxin B1, Deoxynivalenol, and Zearalenone. The standards were sourced from 10 different global market leading manufacturers. To facilitate comparison, all the standard sets were adjusted to the same concentration level. The standards were analyzed using the techniques LC-MS/MS, HPLC-DAD, and LC-HRMS to assess their quality attributes. Regarding the validation of the reference values, it was observed that 30% of the suppliers provided reference standards that were either below the lower acceptance limit or above the higher acceptance limit, confirmed by both the LC-MS/MS and HPLC-DAD methods. Furthermore, a total of 12 impurities were found in the DON standards, 10 in the AFB₁ standards, and 8 in the ZON standards, distributed across all the suppliers. Therefore, this study suggests relevant adjustments to the ISO 17034 standard, proposing that the purity of a raw material should be uniformly based on q-NMR analysis, as most manufacturers state the purity of their certificates is determined using HPLC-UV or LC-MS/MS. Liquid standards with a shelf life of ≤1 year should not exceed an uncertainty of 3%. Standards that have a longer shelf life should not have more than 5% uncertainty. This study also emphasizes the importance of stability. The standards should undergo continuous long-term monitoring; otherwise, products may exhibit a target value of only 80%, as seen in one instance. It is also recommended to include proof of HPLC and LC-MS/MS analyses on the certificate of each released batch of a final product. Full article

(This article belongs to the Section Mycotoxins)

► Show Figures

Figure 1

15 pages, 1683 KiB

Open AccessArticle

Tiny but Mighty: Vipera ammodytes meridionalis (Eastern Long-Nosed Viper) Ontogenetic Venom Variations in Procoagulant Potency and the Impact on Antivenom Efficacies

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

Toxins 2024, 16(9), 396; https://doi.org/10.3390/toxins16090396 - 14 Sep 2024

Viewed by 815

Abstract

The Eastern Long-Nosed Viper (Vipera ammodytes meridionalis) is considered one of the most venomous snakes in Europe. However, it is unknown whether ontogenetic variation in venom effects occurs in this subspecies and how this may impact antivenom efficacy. In this study, [...] Read more.

The Eastern Long-Nosed Viper (Vipera ammodytes meridionalis) is considered one of the most venomous snakes in Europe. However, it is unknown whether ontogenetic variation in venom effects occurs in this subspecies and how this may impact antivenom efficacy. In this study, we compared the procoagulant activities of V. a. meridionalis venom on human plasma between neonate and adult venom phenotypes. We also examined the efficacy of three antivenoms—Viperfav, ViperaTAb, and Inoserp Europe—across our neonate and adult venom samples. While both neonate and adult V. a. meridionalis venoms produced procoagulant effects, the effects produced by neonate venom were more potent. Consistent with this, neonate venom was a stronger activator of blood-clotting zymogens, converting them into their active forms, with a rank order of Factor X >> Factor VII > Factor XII. Conversely, the less potent adult venom had a rank order of FXII marginally more activated than Factor VII, and both much more so than Factor X. This adds to the growing body of evidence that activation of factors besides FII (prothrombin) and FX are significant variables in reptile venom-induced coagulopathy. Although all three examined antivenoms displayed effective neutralization of both neonate and adult V. a. meridionalis venoms, they generally showed higher efficacy on adult venom than on neonate venom. The ranking of antivenom efficacy against neonate venom, from the most effective to the least effective, were Viperfav, Inoserp Europe, ViperaTAb; for adult venom, the ranking was Inoserp Europe, Viperfav, ViperaTAb. Our data reveal ontogenetic variation in V. a meridionalis, but this difference may not be of clinical concern as antivenom was effective at neutralizing both adult and neonate venom phenotypes. Regardless, our results highlight a previously undocumented ontogenetic shift, likely driven by the documented difference in prey preference observed for this species across age classes Full article

(This article belongs to the Special Issue Snake Venom: Toxicology and Associated Countermeasures)

► Show Figures

Figure 1

11 pages, 410 KiB

Open AccessArticle

Assessing the Use of BotulinumtoxinA for Hyperactive Urinary Tract Dysfunction a Decade after Approval: A Single-Blind Study to Evaluate the Reduction in Pain in OnabotulinumtoxinA Detrusor Injection Using Different Injection Needles

by Heinrich Schulte-Baukloh, Catarina Weiss, Thorsten Schlomm, Sarah Weinberger, Hendrik Borgmann, Dirk Höppner, Kathrin Haberecht and Jörg Neymeyer

Toxins 2024, 16(9), 395; https://doi.org/10.3390/toxins16090395 - 14 Sep 2024

Viewed by 348

Abstract

Overactive bladder (OAB) has a significant impact on the quality of life; thus, it requires treatment that can be adhered to over a long period without undue side effects. The current treatment which uses an anticholinergic or β-3 agonist may fail to improve [...] Read more.

Overactive bladder (OAB) has a significant impact on the quality of life; thus, it requires treatment that can be adhered to over a long period without undue side effects. The current treatment which uses an anticholinergic or β-3 agonist may fail to improve symptoms and has side effects, leading to high discontinuation rates. OnabotulinumtoxinA (OnabotA) detrusor injection has been approved for idiopathic OAB as a second-line treatment with good effectiveness and tolerability. This study used a visual analog scale (VAS) to assess the impact of the type of needle used for OnabotA detrusor injections under local anesthesia on the pain levels after each injection. This study included 68 female patients. We used three different needles with thicknesses ranging from 22 to 27 gauge, lengths between 4 and 5 mm, and different cuts. The sensation of pain was rated at each standardized injection location. Regardless of the needle used, the patients’ perceptions of pain at the beginning of the procedure were rated as being less than the subsequent injections. Most pain sensations were rated as low to moderate. The mean pain sensation on the VAS was 2.5 ± 0.3 overall, i.e., for all patients and needles used. Statistically significant differences in pain sensation were rated only at some locations of the bladder (on the back wall and the right side of the bladder). The single needles averaged the following pain scores: 2.8 ± 0.3 for needle A (20 G, 4 mm), 2.1 ± 0.3 for needle B (27 G, 5 mm), and 2.6 ± 0.4 for needle C (20 G, 4 mm, sharp cut 15°). The 27-gauge needle caused significantly less pain, and it had no negative impact due to its length, which was 1 mm longer than the other needles. Thus, the needle thickness was a decisive factor in the patients’ perceptions of pain. Full article

(This article belongs to the Special Issue Botulinum Toxin for Urinary Tract Disease: After a Decade from Approval)

► Show Figures

Figure 1

19 pages, 6415 KiB

Open AccessArticle

Proteome and Phosphoproteome Profiling Reveal the Toxic Mechanism of Clostridium perfringens Epsilon Toxin in MDCK Cells

by Nan Yue, Jing Huang, Mingxin Dong, Jiaxin Li, Shan Gao, Jing Wang, Yingshuang Wang, Dongxue Li, Xi Luo, Tingting Liu, Songyang Han, Lina Dong, Ming Chen, Jinglin Wang, Na Xu, Lin Kang and Wenwen Xin

Toxins 2024, 16(9), 394; https://doi.org/10.3390/toxins16090394 - 14 Sep 2024

Viewed by 521

Abstract

Epsilon toxin (ETX), a potential agent of biological and toxic warfare, causes the death of many ruminants and threatens human health. It is crucial to understand the toxic mechanism of such a highly lethal and rapid course toxin. In this study, we detected [...] Read more.

Epsilon toxin (ETX), a potential agent of biological and toxic warfare, causes the death of many ruminants and threatens human health. It is crucial to understand the toxic mechanism of such a highly lethal and rapid course toxin. In this study, we detected the effects of ETX on the proteome and phosphoproteome of MDCK cells after 10 min and 30 min. A total of 44 differentially expressed proteins (DEPs) and 588 differentially phosphorylated proteins (DPPs) were screened in the 10 min group, while 73 DEPs and 489 DPPs were screened in the 30 min group. ETX-induced proteins and phosphorylated proteins were mainly located in the nucleus, cytoplasm, and mitochondria, and their enrichment pathways were related to transcription and translation, virus infection, and intercellular junction. Meanwhile, the protein–protein interaction network screened out several hub proteins, including SRSF1/2/6/7/11, SF3B1/2, NOP14/56, ANLN, GTPBP4, THOC2, and RRP1B. Almost all of these proteins were present in the spliceosome pathway, indicating that the spliceosome pathway is involved in ETX-induced cell death. Next, we used RNAi lentiviruses and inhibitors of several key proteins to verify whether these proteins play a critical role. The results confirmed that SRSF1, SF3B2, and THOC2 were the key proteins involved in the cytotoxic effect of ETX. In addition, we found that the common upstream kinase of these key proteins was SRPK1, and a reduction in the level of SRPK1 could also reduce ETX-induced cell death. This result was consistent with the phosphorylated proteomics analysis. In summary, our study demonstrated that ETX induces phosphorylation of SRSF1, SF3B2, THOC2, and SRPK1 proteins on the spliceosome pathway, which inhibits normal splicing of mRNA and leads to cell death. Full article

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

► Show Figures

Graphical abstract

22 pages, 2906 KiB

Open AccessArticle

Neutralizing Nanobodies against Venoms from Naja haje Species Captured in North Africa

by Hiba Mejri, Rym Mokrani, Ayoub Ksouri, Mabrouk Seddik, Nour Awad, Gabriel Ayme, Thouraya Chagour, Ahlem Mokrani, Charraf eddine Louchene, Imed Salhi, Rahma Ben Abderrazek, Rym Ben Khalifa, Zakaria Benlasfar, Pierre-Jean Corringer, Mohamed Hammadi, Selma Djilani, Pierre Lafaye and Balkiss Bouhaouala-Zahar

Toxins 2024, 16(9), 393; https://doi.org/10.3390/toxins16090393 - 14 Sep 2024

Viewed by 647

Abstract

Snakebite envenoming (SBE) remains a severely neglected public health issue, particularly affecting tropical and subtropical regions, with Africa experiencing an estimated 435,000 to 580,000 snakebites annually, leading to high morbidity and mortality rates, especially across Africa and Asia. Recognized as a Neglected Tropical [...] Read more.

Snakebite envenoming (SBE) remains a severely neglected public health issue, particularly affecting tropical and subtropical regions, with Africa experiencing an estimated 435,000 to 580,000 snakebites annually, leading to high morbidity and mortality rates, especially across Africa and Asia. Recognized as a Neglected Tropical Disease, SBE management is further complicated by the inadequate efficacy of current antivenom treatments. Of particular concern are cobras (Naja sp.), whose neurotoxins can induce rapid fatal respiratory paralysis. In this study, we investigate the potential of nanobodies as a promising next-generation of immunotherapeutics against cobra venoms. Through a dual strategy of the characterization of venom toxic fractions from cobras captured for the first time in Algeria and Tunisia biotopes, coupled with in vitro assays to evaluate their interactions with acetylcholine receptors, and subsequent immunization of dromedaries to produce specific nanobodies, we identified two lethal fractions, F5 and F6, from each venom, and selected five nanobodies with significant binding and neutralizing of 3DL50 (0.74 mg/kg). The combination of these nanobodies demonstrated a synergistic effect, reaching 100% neutralizing efficacy of 2DL50 lethal venom fraction (0.88 mg/kg) doses in mice. Additionally, our findings highlighted the complex mechanism of cobra venom action through the lethal synergism among its major toxins. Full article

(This article belongs to the Special Issue Passive Immunotherapy and Its Applications in Envenomation and Infectious Diseases: In Tribute to Gaston Ramon (1886–1963))

► Show Figures

Figure 1

19 pages, 528 KiB

Open AccessSystematic Review

Use of Botulinum Toxin in Upper-Limb Tremor: Systematic Review and Perspectives

by Damien Motavasseli, Cécile Delorme, Nicolas Bayle, Jean-Michel Gracies, Emmanuel Roze and Marjolaine Baude

Toxins 2024, 16(9), 392; https://doi.org/10.3390/toxins16090392 - 13 Sep 2024

Viewed by 525

Abstract

Background: Tremor is the most common movement disorder, with significant functional and psychosocial consequences. Oral medications have been disappointing or limited by side effects. Surgical techniques are effective but associated with risks and adverse events. Botulinum toxin (BT) represents a promising avenue but [...] Read more.

Background: Tremor is the most common movement disorder, with significant functional and psychosocial consequences. Oral medications have been disappointing or limited by side effects. Surgical techniques are effective but associated with risks and adverse events. Botulinum toxin (BT) represents a promising avenue but there is still no double-blind evidence of efficacy on upper limb function. A systematic review on the effects of BT in upper-limb tremor was conducted. Methods: A systematic search of the literature was conducted up to July 2023, including the keywords “botulinum toxin” and “tremor”. All randomized controlled trials (RCTs) and open-label studies were analyzed. Independent reviewers assessed their methodological quality. Results: There were only eight published RCTs and seven published open-label studies, with relatively small sample sizes. This review suggests that BT is more effective when injections are patient-tailored, with analyses based on clinical judgement or kinematics. Subjective and objective measures frequently improve but transient weakness may occur after injections, especially if wrist or fingers extensors are targeted. A number of studies had methodological limitations. Conclusions: The authors discuss how to optimize tremor assessments and effects of BT injection. Controlled evidence is still lacking but it is suggested that distal “asymmetric” BT injections (targeting flexors/pronators while sparing extensors/supinators) and proximal injections, involving shoulder rotators when indicated, may avoid excessive weakness while optimizing functional benefit. Full article

(This article belongs to the Special Issue Application of Botulinum Toxins in Non-spastic and Neurodegenerative Disorders)

► Show Figures

Figure 1

13 pages, 1367 KiB

Open AccessArticle

An Ultrasonographic Analysis of the Deep Inferior Tendon in the Masseter Muscle: Implications for Botulinum Toxin Injections

by Jia Shi, Chenyang Li, Jinbo Zhou, Xinyu Guo, Guo Li and Meng You

Toxins 2024, 16(9), 391; https://doi.org/10.3390/toxins16090391 - 11 Sep 2024

Viewed by 381

Abstract

(1) Background: With the increasing aesthetic pursuit of facial features, the clinical use of Botulinum Toxin Type A (BoNT-A) injections for masseter hypertrophy has been on the rise. However, due to variations in masseter muscle structure and differences in clinicians’ injection techniques, blind [...] Read more.

(1) Background: With the increasing aesthetic pursuit of facial features, the clinical use of Botulinum Toxin Type A (BoNT-A) injections for masseter hypertrophy has been on the rise. However, due to variations in masseter muscle structure and differences in clinicians’ injection techniques, blind injections may lack precision, potentially compromising treatment accuracy and increasing the risk of complications. (2) Objectives: The study aims to use ultrasonography to detail the deep inferior tendon (DIT) within the masseter muscle in a young Chinese cohort, refine its classification, analyze muscle belly thickness and variations across groups, and propose a customized ultrasound-guided BoNT-A injection protocol. (3) Methods: Ultrasound imaging was used to observe the bilateral masseter muscles at rest and during clenching. The features of the DIT were classified from these images, and the thickness of the masseter’s distinct bellies associated with the DIT types was measured in both states. (4) Results: The study cohort included 103 participants (27 male, 76 female), with 30 muscles in the normal masseter group and 176 muscles in the hypertrophy group. The DIT was categorized as Type A, B (subtypes B1, B2), and C. The distribution of these types was consistent across normal, hypertrophic, and gender groups, all following the same trend (B > A > C). In hypertrophy, Type B1 showed uniform thickness across masseter bellies, B2 presented with a thinner intermediate belly, and Type C had mainly superficial muscle enlargement. Changes in muscle thickness during clenching were noted but not statistically significant among different bellies. (5) Conclusions: The study evidences individual variation in the DIT, highlighting the importance of precise DIT classification for effective BoNT-A injections. A tailored ultrasound-guided BoNT-A injection strategy based on this classification may enhance safety and efficacy of the therapy. Full article

► Show Figures

Figure 1

30 pages, 1027 KiB

Open AccessArticle

Stability of Resistance of Maize to Ear Rots (Fusarium graminearum, F. verticillioides and Aspergillus flavus) and Their Resistance to Toxin Contamination and Conclusions for Variety Registration

by Akos Mesterhazy, Balazs Szabo, Denes Szieberth, Szabolcs Tóth, Zoltan Nagy, Tamas Meszlenyi, Beata Herczig, Attila Berenyi and Beata Tóth

Toxins 2024, 16(9), 390; https://doi.org/10.3390/toxins16090390 - 10 Sep 2024

Viewed by 629

Abstract

All major ear rots (F. graminearum, F. verticillioides, and Aspergillus flavus) and their toxins are present in maize of preharvest origin in Hungary. Resistance can be an important tool in reducing the infection and toxin contamination from these rots [...] Read more.

All major ear rots (F. graminearum, F. verticillioides, and Aspergillus flavus) and their toxins are present in maize of preharvest origin in Hungary. Resistance can be an important tool in reducing the infection and toxin contamination from these rots in maize. Previous results identified resistance differences in maize hybrids that were suitable for use in evaluating their risk from toxigenic fungi and their toxins. During the tests, two methodical improvements were achieved: the use of three isolates of the fungus secured and a more precise estimation of resistance to ear rots and their resistance to toxin accumulation or overproduction. The improvement in sampling and the tests of subsamples made the evaluation for the statistics much more exact. This way, we were able to reduce the Within value, providing a statistically more reliable method of evaluation. Earlier data had confirmed that toxin contamination could not be predicted well from visual ear rot severity data. Contradictory results for hybrid ranking were often identified between isolates. The resistance to disease and toxin contamination is not generally valid. The new suggested methodology compares the performance of hybrids in a large number of epidemic situations to identify adaptable hybrids that can respond to diverse conditions; therefore, the stability of resistance and toxin response is decisive information to evaluate risk analyses. The increased number of disease toxin data allowed for lower LSD 5% values for toxins, a much finer analysis of toxin overproduction and underproduction, and a wider database for stability analyses. This way, we obtained important additional separated information about resistance to accumulation of toxins and about maize resistance to these pathogens that is suitable to provide much more reliable testing than was possible until now. Globally, about 50–100 million metric tons can be saved by excluding susceptible hybrids from commercial production. Full article

(This article belongs to the Special Issue Effect of Mycotoxins on Crops and Their Prevention)

► Show Figures

Figure 1

16 pages, 37816 KiB

Open AccessArticle

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

Viewed by 420

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)

► Show Figures

Graphical abstract

10 pages, 265 KiB

Open AccessArticle

Do Diagnostic Nerve Blocks Affect the Starting Dose of Botulinum Neurotoxin Type A for Spasticity? A Case-Control Study

by Mirko Filippetti, Stefano Tamburin, Rita Di Censo, Roberto Aldegheri, Elisa Mantovani, Stefania Spina, Marco Battaglia, Alessio Baricich, Andrea Santamato, Nicola Smania and Alessandro Picelli

Toxins 2024, 16(9), 388; https://doi.org/10.3390/toxins16090388 - 6 Sep 2024

Viewed by 827

Abstract

One of the aims of diagnostic nerve blocks is to identify the overactive muscles that lead to a specific spasticity pattern. However, to date, there is no evidence on how nerve blocks may affect botulinum neurotoxin-A (BoNT-A) dose in patients with spasticity. This [...] Read more.

One of the aims of diagnostic nerve blocks is to identify the overactive muscles that lead to a specific spasticity pattern. However, to date, there is no evidence on how nerve blocks may affect botulinum neurotoxin-A (BoNT-A) dose in patients with spasticity. This case-control study aims to assess the role of diagnostic nerve block in defining BoNT-A starting dose at first treatment. Patients with upper and lower limb spasticity treated for the first time with BoNT-A were retrospectively divided into two groups: Group 1 (n = 43) was evaluated with clinical assessment and diagnostic nerve block; Group 2 (n = 56) underwent clinical assessment only. Group 1 was injected with higher BoNT-A doses in some muscles (i.e., flexor digitorum profundus, soleus), and received a higher BoNT-A cumulative dose with a larger number of injected muscles for some spasticity patterns (i.e., “clenched fist”, “flexed fingers”, “adducted thigh”). Diagnostic nerve block may help the clinician to optimize and personalize the BoNT-A dose since the first BoNT-A treatment. Full article

(This article belongs to the Special Issue The Botulinum Toxin and Spasticity: Exploring New Horizons)

18 pages, 18199 KiB

Open AccessArticle

Comparison of Neural Recovery Effects of Botulinum Toxin Based on Administration Timing in Sciatic Nerve-Injured Rats

by Minsu Seo, Seokjoon Hwang, Tae Heon Lee and Kiyeun Nam

Toxins 2024, 16(9), 387; https://doi.org/10.3390/toxins16090387 - 5 Sep 2024

Viewed by 462

Abstract

This study aimed to assess the effects of the timing of administering botulinum neurotoxin A (BoNT/A) on nerve regeneration in rats. Sixty 6-week-old rats with a sciatic nerve injury were randomly divided into four groups: the immediately treated (IT) group (BoNT/A injection administered [...] Read more.

This study aimed to assess the effects of the timing of administering botulinum neurotoxin A (BoNT/A) on nerve regeneration in rats. Sixty 6-week-old rats with a sciatic nerve injury were randomly divided into four groups: the immediately treated (IT) group (BoNT/A injection administered immediately post-injury), the delay-treated (DT) group (BoNT/A injection administered one week post-injury), the control group (saline administered one week post-injury), and the sham group (only skin and muscle incisions made). Nerve regeneration was assessed 3, 6, and 9 weeks post-injury using various techniques. The levels of glial fibrillary acid protein (GFAP), astroglial calcium-binding protein S100β (S100β), growth-associated protein 43 (GAP43), neurofilament 200 (NF200), and brain-derived neurotrophic factor (BDNF) in the IT and DT groups were higher. ELISA revealed the highest levels of these proteins in the IT group, followed by the DT and control groups. Toluidine blue staining revealed that the average area and myelin thickness were higher in the IT group. Electrophysiological studies revealed that the CMAP in the IT group was significantly higher than that in the control group, with the DT group exhibiting significant differences starting from week 8. The findings of the sciatic functional index analysis mirrored these results. Thus, administering BoNT/A injections immediately after a nerve injury is most effective for neural recovery. However, injections administered one week post-injury also significantly enhanced recovery. BoNT/A should be administered promptly after nerve damage; however, its administration during the non-acute phase is also beneficial. Full article

(This article belongs to the Special Issue Botulinum Toxins: New Uses in the Treatment of Diseases (Volume II))

► Show Figures

Figure 1

11 pages, 777 KiB

Open AccessArticle

Predictive Factors for the Successful Outcome of Urethral Sphincter Injections of Botulinum Toxin A for Non-Neurogenic Dysfunctional Voiding in Women

by Chia-Cheng Yang, Yuan-Hong Jiang and Hann-Chorng Kuo

Toxins 2024, 16(9), 386; https://doi.org/10.3390/toxins16090386 - 5 Sep 2024

Viewed by 406

Abstract

Purpose: Dysfunctional voiding (DV) is not uncommon in women with non-neurogenic voiding dysfunction. Because of its unknown pathophysiology, effective and durable treatment is lacking. This study aimed to analyze the results of treatment and predictive factors for a successful outcome of botulinum toxin [...] Read more.

Purpose: Dysfunctional voiding (DV) is not uncommon in women with non-neurogenic voiding dysfunction. Because of its unknown pathophysiology, effective and durable treatment is lacking. This study aimed to analyze the results of treatment and predictive factors for a successful outcome of botulinum toxin A (BoNT-A) treatment in female patients with DV. Methods: In total, 66 women with DV confirmed by a videourodynamic study (VUDS) were treated with a BoNT-A injection into the urethral sphincter once (n = 33) or several times (n = 33). VUDS was performed before (baseline) and after the BoNT-A treatment. Patients with a global response assessment of the voiding condition of 2 or 3 and a voiding efficiency (VE) of >20% than baseline were considered to have a successful outcome. The baseline demographics, VUDS parameters, and VUDS DV subtypes were compared between the successful and failed groups. Predictive factors for a successful outcome were investigated by logistic regression analyses. Results: Successful and failed outcomes were achieved in 27 (40.9%) and 39 (59.1%) women, respectively. After BoNT-A injections, the maximum flow rate (Qmax), voided volume, and VE all significantly increased, and the postvoid residual (PVR) was slightly improved. No significant difference in the number of injections and medical comorbidity was found between the groups. However, the successful group had a higher incidence of previous pelvic surgery. No significant difference in the treatment outcome was found among patients with different urethral obstruction sites. Significant improvements in Qmax, voided volume, PVR, VE, and the bladder outlet obstruction (BOO) index were noted in the successful group. A lower VE at baseline and a history of surgery were identified as predictive factors for a successful outcome of BoNT-A injections for treating DV. Conclusion: BoNT-A injections into the urethral sphincter can effectively improve VE in 40.9% of women with DV. Women with higher BOO grades and previous pelvic surgery are predicted to have a successful treatment outcome. Full article

(This article belongs to the Section Bacterial Toxins)

► Show Figures

Figure 1

12 pages, 1299 KiB

Open AccessArticle

Rapid Detection of Aflatoxins in Ground Maize Using Near Infrared Spectroscopy

by Sylviane Bailly, Béatrice Orlando, Jean Brustel, Jean-Denis Bailly and Cecile Levasseur-Garcia

Toxins 2024, 16(9), 385; https://doi.org/10.3390/toxins16090385 - 4 Sep 2024

Viewed by 618

Abstract

Aflatoxins are carcinogenic mycotoxins that may contaminate many crops and more especially maize. To protect consumers from these contaminants, many countries set up low regulatory thresholds of few µg/kg. The control of food requires time-consuming analysis for which sampling is a key step. [...] Read more.

Aflatoxins are carcinogenic mycotoxins that may contaminate many crops and more especially maize. To protect consumers from these contaminants, many countries set up low regulatory thresholds of few µg/kg. The control of food requires time-consuming analysis for which sampling is a key step. It would therefore of key sanitary and economic relevance to develop rapid, sensitive and accurate methods that could even be applied on line at harvest, to identify batches to be excluded as soon as possible. In this study, we analyzed more than 500 maize samples taken at harvest during 3 years for their aflatoxin contamination using HPLC-MS. Among them, only 7% were contaminated but sometimes at levels largely exceeding European regulations. We demonstrate that Near InfraRed Spectroscopy (NIRS) could be of great help to classify cereal samples according to their level of aflatoxin contamination (below or higher than E.U. regulation). To build the model, all AF contaminated samples as well as an equivalent number of AF free samples were used. NIRS performance was not sufficient to quantify the toxins with adequate precision. However, its ability to discriminate naturally contaminated maize samples according to their level of contamination with aflatoxins in relation to European regulations using a quadratic PCA-DA model was excellent. Accuracy of the model was 97.4% for aflatoxin B1 and 100% for total aflatoxins. Full article

(This article belongs to the Special Issue Editorial Board Members’ Collection Series: Advances in Rapid Mycotoxin Testing)

► Show Figures

Figure 1

15 pages, 1537 KiB

Open AccessArticle

Utility of Cry1Ja for Transgenic Insect Control

by John P. Mathis, Catherine Clark, Amit Sethi, Benchie Ortegon, Gilda Rauscher, Russ Booth, Samuel Coder and Mark E. Nelson

Toxins 2024, 16(9), 384; https://doi.org/10.3390/toxins16090384 - 4 Sep 2024

Viewed by 465

Abstract

Insect control traits are a key component of improving the efficacy of insect pest management and maximizing crop yields for growers. Insect traits based on proteins expressed by the bacteria Bacillus thuringiensis (Bt) have proven to be very effective tools in achieving this [...] Read more.

Insect control traits are a key component of improving the efficacy of insect pest management and maximizing crop yields for growers. Insect traits based on proteins expressed by the bacteria Bacillus thuringiensis (Bt) have proven to be very effective tools in achieving this goal. Unfortunately, the adaptability of insects has led to resistance to certain proteins in current commercial products. Therefore, new insecticidal traits representing a different mode of action (MoA) than those currently in use are needed. Cry1Ja has good insecticidal activity against various lepidopteran species, and it provides robust protection against insect feeding with in planta expression. For Bt proteins, different MoAs are determined by their binding sites in the insect midgut. In this study, competitive binding assays are performed using brush border membrane vesicles (BBMVs) from Helicoverpa zea, Spodoptera frugiperda, and Chrysodeixis includens to evaluate the MoA of Cry1Ja relative to representatives of the various Bt proteins that are expressed in current commercial products for lepidopteran insect protection. This study highlights differences in the shared Cry protein binding sites in three insect species, Cry1Ja bioactivity against Cry1Fa resistant FAW, and in planta efficacy against target pests. These data illustrate the potential of Cry1Ja for new insect trait development. Full article

(This article belongs to the Section Bacterial Toxins)

► Show Figures

Figure 1

14 pages, 2647 KiB

Open AccessArticle

Long-Term Management of Post-Stroke Spasticity with Botulinum Toxin: A Retrospective Study

by Nicoletta Falcone, Fabrizio Leo, Carmelo Chisari and Stefania Dalise

Toxins 2024, 16(9), 383; https://doi.org/10.3390/toxins16090383 - 3 Sep 2024

Viewed by 643

Abstract

Stroke-induced spasticity is a prevalent condition affecting stroke survivors, significantly impacting their quality of life. Botulinum Toxin A injections are widely used for its management, yet the long-term effects and optimal management strategies remain uncertain. This retrospective study analyzed medical records of 95 [...] Read more.

Stroke-induced spasticity is a prevalent condition affecting stroke survivors, significantly impacting their quality of life. Botulinum Toxin A injections are widely used for its management, yet the long-term effects and optimal management strategies remain uncertain. This retrospective study analyzed medical records of 95 chronic stroke patients undergoing long-term BoNT-A treatment for spasticity. Demographic data, treatment duration, dosage variability, and dropout rates were assessed over a period ranging from 2 to 14 years. The study revealed a notable extension of the interval between BoNT-A injections throughout the treatment duration. Dropout rates peaked during the initial 5 years of treatment, perhaps due to perceived treatment ineffectiveness. Additionally, a trend of escalating dosage was observed across all groups, indicating a potential rise in the severity of spasticity or changes in treatment response over time. BoNT-A injections emerged as the predominant treatment choice for managing post-stroke spasticity. The delayed initiation of BoNT-A treatment underscores the need for heightened awareness among healthcare providers to recognize and manage spasticity promptly post-stroke. Patients’ expectations and treatment goals should be clearly defined to optimize treatment adherence, while the observed escalation in dosage and treatment intervals emphasizes the dynamic nature of spasticity and underscores the importance of monitoring long-term treatment outcomes. Full article

(This article belongs to the Section Bacterial Toxins)

► Show Figures

Figure 1

12 pages, 1615 KiB

Open AccessArticle

The Mycotoxins T-2 and Deoxynivalenol Facilitate the Translocation of Streptococcus suis across Porcine Ileal Organoid Monolayers

by Xiaonan Guan, Arabela R. Martinez, Marcela Fernandez, Francesc Molist, Jerry M. Wells and Regiane R. Santos

Toxins 2024, 16(9), 382; https://doi.org/10.3390/toxins16090382 - 1 Sep 2024

Viewed by 825

Abstract

Mycotoxins have the potential to increase the risk of airway or intestinal infection due to their effects on epithelial integrity and function. The bacterium Streptococcus suis (S. suis) is often carried in pigs and can cause outbreaks of invasive disease, leading to sepsis [...] Read more.

Mycotoxins have the potential to increase the risk of airway or intestinal infection due to their effects on epithelial integrity and function. The bacterium Streptococcus suis (S. suis) is often carried in pigs and can cause outbreaks of invasive disease, leading to sepsis and meningitis in postweaning piglets. In this study, we tested the effect of two Fusarium mycotoxins (deoxynivalenol (DON) and T-2) on the integrity of the intestinal epithelium and their interaction with S. suis. Porcine ileal organoids were exposed to DON and T-2 individually or in combination and co-cultured with or without S. suis. Both DON and T-2 were toxic for ileal organoid monolayers at a concentration of 1 µM but not S. suis, even at a higher concentration of 4 µM. To mimic sub-clinical exposures on farms, DON was tested at a concentration of 0.1 µM and T-2 at a concentration of 0.01 µM. The mycotoxins alone did not affect cell permeability, but in combination with S. suis there was an increase in epithelial permeability. Furthermore, DON and T-2 together decreased the transepithelial electrical resistance and increased bacterial translocation. Full article

(This article belongs to the Section Mycotoxins)

► Show Figures

Figure 1

14 pages, 4930 KiB

Open AccessArticle

Apoptosis and Oxidative Stress in Human Intestinal Epithelial Caco-2 Cells Caused by Marine Phycotoxin Azaspiracid-2

by Liye Zhao, Jiangbing Qiu, Jingrui Zhang, Aifeng Li and Guixiang Wang

Toxins 2024, 16(9), 381; https://doi.org/10.3390/toxins16090381 - 31 Aug 2024

Viewed by 581

Abstract

When humans consume seafood contaminated by lipophilic polyether phycotoxins, such as azaspiracids (AZAs), the toxins are mainly leached and absorbed in the small intestine, potentially causing intestinal damage. In this study, human intestinal epithelial Caco-2 cells were used to investigate the adverse effects [...] Read more.

When humans consume seafood contaminated by lipophilic polyether phycotoxins, such as azaspiracids (AZAs), the toxins are mainly leached and absorbed in the small intestine, potentially causing intestinal damage. In this study, human intestinal epithelial Caco-2 cells were used to investigate the adverse effects of azaspiracid-2 (AZA-2) on human intestinal epithelial cells. Cell viability, apoptosis, oxidative damage and mitochondrial ultrastructure were investigated, and ribonucleic acid sequence (RNA-seq) analysis was applied to explore the potential mechanisms of AZA-2 toxicity to Caco-2 cells. Results showed that AZA-2 significantly reduced the proliferation of Caco-2 cells in a concentration-dependent response, and the 48 h EC₅₀ of AZA-2 was 12.65 nmol L⁻¹. AZA-2 can induce apoptosis in Caco-2 cells in a dose-dependent manner. Visible mitochondrial swelling, cristae disintegration, membrane rupture and autophagy were observed in Caco-2 cells exposed to AZA-2. Reactive oxygen species (ROS) and malondialdehyde (MDA) content were significantly increased in Caco-2 cells after 48 h of exposure to 1 and 10 nmol L⁻¹ of AZA-2. Transcriptome analysis showed that KEGG pathways related to cellular oxidative damage and lipid metabolism were affected, mainly including mitophagy, oxidative phosphorylation, cholesterol metabolism, vitamin digestion and absorption, bile secretion and the peroxisome proliferator-activated receptor signaling pathway. The cytotoxic effects of AZA-2 on Caco-2 cells may be associated with ROS-mediated autophagy and apoptosis in mitochondrial cells. Results of this study improve understanding of the cytotoxicity and molecular mechanisms of AZA-2 on Caco-2 cells, which is significant for protecting human health. Full article

► Show Figures

Figure 1

Journal Menu

Journal Browser

Toxins, Volume 16, Issue 9 (September 2024) – 37 articles

Further Information

Guidelines

MDPI Initiatives

Follow MDPI