Toxins, Volume 15, Issue 10 (October 2023) – 36 articles

Karenia mikimotoi is a common species of red tide dinoflagellate that causes the mass mortality of marine fauna in coastal waters of Republic of Korea. Despite continuous studies on the ecophysiology and toxicity of K. mikimotoi, the underlying molecular mechanisms remain poorly understood. Red sea bream, Pagrus major, is a high-value aquaculture fish species, and the coastal aquaculture industry of red sea bream has been increasingly affected by red tides. To investigate the potential oxidative effects of K. mikimotoi on P. major and the molecular mechanisms involved, we exposed the fish to varying concentrations of K. mikimotoi and evaluated its toxicity. Our results showed that exposure to K. mikimotoi led to an accumulation of reactive oxygen species (ROS) and oxidative DNA damage in the gill tissue of P. major. Furthermore, we found that K. mikimotoi induced the activation of antioxidant enzymes, such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, in the gill tissue of P. major, with a significant increase in activity at concentrations above 5000 cells/mL. However, the activity of glutathione S-transferase did not significantly increase at the equivalent concentration. Our study confirms that oxidative stress and DNA damage is induced by acute exposure to K. mikimotoi, as it produces ROS and hypoxic conditions in P. major. In addition, it was confirmed that gill and blood samples can be used as biomarkers to detect the degree of oxidative stress in fish. These findings have important implications for the aquaculture of red sea bream, particularly in the face of red tide disasters. Full article

Despite more than 80 years of use in a number of conditions, including in critically ill patients, comments have recently arisen regarding the safety and efficacy of human serum albumin (HSA) as a therapeutic product and stabilizer/excipient in botulinum neurotoxins. This review summarizes the literature on the safety of HSA. Beyond decades of safe use, the largest clinical dataset of HSA safety is a large meta-analysis of HSA supplier data, which found only an extremely remote risk of serious adverse events across millions of doses of therapeutic concentrations of HSA. There is a paucity of literature identifying HSA-specific adverse events when used as a stabilizer/excipient; however, studies of HSA-containing botulinum neurotoxins (BoNTs) suggest that adverse events are not related to HSA. Polysorbates, which are synthetically produced and not physiologically inert, are contained in pending or new-to-market BoNT formulations. In contrast to HSA, evidence exists to suggest that polysorbates (particularly PS20/PS80) can cause serious adverse events (e.g., hypersensitivity, anaphylaxis, and immunogenicity). Full article

Background: The objective of this study was to provide evidence from a simple simulation. In patients with focal dystonia, an initial good response to botulinum neurotoxin (BoNT) injections followed by a secondary worsening does not necessarily arise from an antibody-induced secondary treatment failure (NAB-STF), but may stem from a “pseudo”-secondary treatment failure (PSEUDO-STF). Methods: The simulation of the outcome after BoNT long-term treatment was performed in four steps: 1. The effect of the first single BoNT injection (SI curve) was displayed as a 12-point graph, corresponding to the mean improvement from weeks 1 to 12. 2. The remaining severity of the dystonia during the nth injection cycle was calculated by subtracting the SI curve (weighted by the outcome after n − 1 cycles) from the outcome after week 12 of the (n − 1)th cycle. 3. A graph was chosen (the PRO curve), which represents the progression of the severity of the underlying disease during BoNT therapy. 4. The interaction between the outcome during the nth BoNT cycle and the PRO curve was determined. Results: When the long-term outcome after n cycles of BoNT injections (applied every 3 months) was simulated as an interactive process, subtracting the effect of the first cycle (weighted by the outcome after n − 1 cycles) and adding the progression of the disease, an initial good improvement followed by secondary worsening results. This long-term outcome depends on the steepness of the progression and the duration of action of the first injection cycle. We termed this response behavior a “pseudo”-secondary treatment failure, as it can be compensated via a dose increase. Conclusion: A secondary worsening following an initial good response in BoNT therapy of focal dystonia might not necessarily indicate neutralizing antibody induction but could stem from a “PSEUDO”-STF (a combination of good response behavior and progression of the underlying disease). Thus, an adequate dose adaptation must be conducted before diagnosing a secondary treatment failure in the strict sense. Full article

Zearalenone (ZEN), a non-steroidal estrogenic fungal toxin widely present in forage, food, and their ingredients, poses a serious threat to animal and human reproductive health. ZEN also threatens ovine, a major source of human food and breeding stock. However, the mechanisms underlying the impact of ZEN on the in vitro maturation (IVM) of ovine oocytes remain unclear. This study aimed to elucidate these mechanisms using the Smart-seq2 technology. A total of 146 differentially expressed genes were obtained, using Smart-seq2, from sheep oocytes cultured in vitro after ZEN treatment. ZEN treatment inhibited RUNX2 and SPP1 expression in the PI3K signaling pathway, leading to the downregulation of THBS1 and ultimately the downregulation of TNFAIP6; ZEN can also decrease TNFAIP6 by reducing PTPRC and ITGAM. Both inhibit in vitro maturation of ovine oocytes and proliferation of cumulus cells by downregulating TNFAIP6. These findings provide data and a theoretical basis for elucidating ZEN’s toxicity mechanisms, screening therapeutic drugs, and reducing ZEN-related losses in the ovine industry. Full article

Macrophage plasticity is a fundamental feature of the immune response since it favors the rapid and adequate change of the functional phenotype in response to the pathogen or the microenvironment. Several studies have shown that Crotoxin (CTX), the major toxin of the Crotalus durissus terrificus snake venom, has a long-lasting antitumor effect both in experimental models and in clinical trials. In this study, we show the CTX effect on the phenotypic reprogramming of macrophages in the mesenchymal tumor microenvironment or those obtained from the peritoneal cavity of healthy animals. CTX (0.9 or 5 μg/animal subcutaneously) administered concomitantly with intraperitoneal inoculation of tumor cells (1 × 10⁷/0.5 mL, injected intraperitoneally) of Ehrlich Ascitic Tumor (EAT) modulated the macrophages phenotype (M1), accompanied by increased NO^• production by cells from ascites, and was evaluated after 13 days. On the other hand, in healthy animals, the phenotypic profile of macrophages was modulated in a dose-dependent way at 0.9 μg/animal: M1 and at 5.0 μg/animal: M2; this was accompanied by increased NO^• production by peritoneal macrophages only for the dose of 0.9 μg/animal of CTX. This study shows that a single administration of CTX interferes with the phenotypic reprogramming of macrophages, as well as with the secretory state of cells from ascites, influencing events involved with mesenchymal tumor progression. These findings may favor the selection of new therapeutic targets to correct compromised immunity in different systems. Full article

Wheat-based products are largely consumed by children worldwide. Deoxynivalenol (DON) is known for its acute and chronic toxicity and is the most common contaminant of cereal grains. Since no legal limits are set for DON in wheat-based products and specific foods intended for children over 3 years on the market, a high risk of overexposure to this contaminant may emerge. The main objective of the study, conducted in 2018–2019, was to produce a wheat flour intended for children over three years, characterized by a high level of safety in terms of DON content, to be used to produce wheat-derived products. The dedicated flour was produced by adopting tailored procedures like the selection of wheat suppliers, the predetermination of the safe contamination of DON in the final products, and the evaluation of the transfer rate from the wheat flour to derived products (bread, breadsticks, biscuits, plumcake, and focaccia). The results showed that the daily exposure of children was considered to be safe, in a range between 7% (biscuits) and 67% (bread) of DON tolerable daily intake (TDI) and that only by producing a flour characterized by DON levels much lower than those in force, can “safe” products be marketed. Full article

Bothrops venoms are rich in enzymes acting on platelets and coagulation. This action is dependent on two major co-factors, i.e., calcium and phospholipids, while antivenoms variably neutralize venom-related coagulopathy effects. Our aims were (i) to describe the composition of B. atrox and B. lanceolatus venoms; (ii) to study their activity on the whole blood using rotational thromboelastometry (ROTEM); (iii) to evaluate the contribution of calcium and phospholipids in their activity; and (iv) to compare the effectiveness of four antivenoms (Bothrofav^™, Inoserp^™ South America, Antivipmyn^™ TRI, and PoliVal-ICP^™) on the procoagulant activity of these two venoms. Venom composition was comparable. Both venoms exhibited hypercoagulant effects. B. lanceolatus venom was completely dependent on calcium but less dependent on phospholipids than B. atrox venom to induce in vitro coagulation. The four antivenoms neutralized the procoagulant activity of the two venoms; however, with quantitative differences. Bothrofav^™ was more effective against both venoms than the three other antivenoms. The relatively similar venom-induced effects in vitro were unexpected considering the opposite clinical manifestations resulting from envenomation (i.e., systemic bleeding with B. atrox and thrombosis with B. lanceolatus). In vivo studies are warranted to better understand the pathophysiology of systemic bleeding and thrombosis associated with Bothrops bites. Full article

Optimisation of solar drying to reduce fungal growth and Ochratoxin A (OTA) contamination is a crucial concern in raisin and currant production. Stochastic and deterministic analysis has been utilized to investigate environmental indicators and drying characteristics. The analysis was performed using two seedless grape varieties (Crimson—red and Thompson—white) that were artificially inoculated with Aspergillus carbonarius during open-air and tunnel drying. Air temperature (T) and relative humidity (RH) were measured and analysed during the drying experiment, along with grape surface temperature (T_s), and water activity (a_w). The grape moisture content, fungal colonization, and OTA contamination were estimated, along with the water diffusivity (D_eff) and peel resistance (r_peel) to water transfer. Monitoring the surface temperature of grapes is essential in the early detection of fungal growth and OTA contamination. As surface temperature should be carried out continuously, remote sensing protocols, such as infrared sensors, provide the most efficient means to achieve this. Furthermore, data collection and analysis could be conducted through the Internet of Things (IoT), thereby enabling effortless accessibility. The average T_s of the grapes was 6.5% higher in the tunnel than in the open-air drying. The difference between the RH of air and that in the plastic crates was 16.26–17.22%. In terms of CFU/mL, comparison between white and red grapes in the 2020 and 2021 experiments showed that the red grapes exhibited significantly higher values than the white grapes. Specifically, the values for red grapes were 4.3 in 2021 to 3.4 times in 2020 higher compared to the white grapes. On the basis of the conducted analysis, it was concluded that tunnel drying provided some advantages over open-air drying, provided that hygienic and managerial requirements are met. Full article

Cancer progression is characterized by microenvironmental acidification. Tumor cells adapt to low environmental pH by activating acid-sensing trimeric ion channels of the DEG/ENaC family. The α-ENaC/ASIC1a/γ-ENaC heterotrimeric channel is a tumor-specific acid-sensing channel, and its targeting can be considered a new strategy for cancer therapy. Mambalgin-2 from the Dendroaspis polylepis venom inhibits the α-ENaC/ASIC1a/γ-ENaC heterotrimer more effectively than the homotrimeric ASIC1a channel, initially proposed as the target of mambalgin-2. Although the molecular basis of such mambalgin selectivity remained unclear. Here, we built the models of the complexes of mambalgin-2 with the α-ENaC/ASIC1a/γ-ENaC and ASIC1a channels, performed MD and predicted the difference in the binding modes. The importance of the ‘head’ loop region of mambalgin-2 for the interaction with the hetero-, but not with the homotrimeric channel was confirmed by site-directed mutagenesis and electrophysiology. A new mode of allosteric regulation of the ENaC channels by linking the thumb domain of the ASIC1a subunit with the palm domain of the γ-ENaC subunit was proposed. The data obtained provide new insights into the regulation of various types of acid-sensing ion channels and the development of new strategies for cancer treatment. Full article

Cyanobacteria harmful algal blooms (cyanoHABs) can have impacts on human health, aquatic ecosystems, and the economy. Nutrient management is an important mitigation and even remediation strategy. In this work, the paralytic shellfish toxin (PST)-producing Aphanizomenon (Aphan.) flos-aquae (Linnaeus) Ralfs ex Bornet & Flahault (now identified as Aphan. sp.) single filament isolate NH-5 was grown in P-depleted media, N-depleted media, and complete BG-11 media. Growth and heterocyst and vegetative cells were monitored using dry weight and cell counts. Ultrasonication was used to separate heterocysts from vegetative cells. HPLC-FLD with post-column derivatization was used to determine the saxitoxin (STX) and neosaxitoxin (NEOSTX) concentration per cell. Aphan. sp. NH-5 biomass was lower in the P-depleted media than in the N-depleted media and the control, though higher heterocyst counts were detected in the N-depleted media. The heterocyst toxin concentration was significantly higher compared to the vegetative cells for the N-depleted media, control, and P-depleted media. However, no significant differences were found among all preparations with regard to the STX-to-NEOSTX ratio. We conclude that N limitation induced higher heterocyst numbers and that N fixation activity is a factor behind the increase in the STX and NEOSTX production of Aphan. sp. NH-5. Full article

Mycotoxin contamination on food and feed can have deleterious effect on human and animal health. Agricultural crops may contain one or more mycotoxin compounds; therefore, a good multiplex detection method is desirable to ensure food safety. In this study, we developed a rapid method using label-free surface-enhanced Raman spectroscopy (SERS) to simultaneously detect three common types of mycotoxins found on corn, namely aflatoxin B1 (AFB1), zearalenone (ZEN), and ochratoxin A (OTA). The intrinsic chemical fingerprint from each mycotoxin was characterized by their unique Raman spectra, enabling clear discrimination between them. The limit of detection (LOD) of AFB1, ZEN, and OTA on corn were 10 ppb (32 nM), 20 ppb (64 nM), and 100 ppb (248 nM), respectively. Multivariate statistical analysis was used to predict concentrations of AFB1, ZEN, and OTA up to 1.5 ppm (4.8 µM) based on the SERS spectra of known concentrations, resulting in a correlation coefficient of 0.74, 0.89, and 0.72, respectively. The sampling time was less than 30 min per sample. The application of label-free SERS and multivariate analysis is a promising method for rapid and simultaneous detection of mycotoxins in corn and may be extended to other types of mycotoxins and crops. Full article

Daboia (Vipera) palaestinae (Dp), accounts for most envenomations in humans and dogs in Israel. In humans envenomed by Dp, serum cholesterol concentration (sChol) is inversely correlated with envenomation severity. This study examined the utility of sChol upon admission in dogs envenomed by Dp as an envenomation severity and outcome marker. Data upon admission, including sChol, were retrospectively collected from the medical records of dogs with proven Dp envenomation. The study included 415 dogs. The mortality rate was 11%. The heart rate upon admission was higher in non-survivors than in survivors. Signs of bleeding or hematoma and circulatory shock signs were more frequent among non-survivors compared to survivors. sChol, the platelet count, and serum albumin concentration (sAlb) were lower, while serum creatinine concentration was higher among non-survivors. sChol and sAlb were moderately, positively, and significantly correlated. sChol was significantly, negatively, albeit weakly, correlated with the length of hospitalization and the heart rate. sChol was lower in dogs admitted >12 h post-envenomation than in those admitted later. In dogs, sChol upon admission is a potential marker of severity and outcome of Dp envenomation. The platelet count, sAlb, and sCreat might also be potential markers. Full article

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

The present article focuses on a strain of ascomycete GF3 isolated from a water sample taken in the Gulf of Finland. Based on phylogenetic analysis data, the isolate was identified as Penicillium sp. GF3. The fungus GF3 demonstrates algicidal activity towards cyanobacteria (98–100%). The algicidal effect on green algae did not exceed 50%. The isolate GF3 exhibits an indirect attack mode by releasing metabolites with algicidal and/or lytic activity into the environment. Moreover, the strain Penicillium sp. GF3 is able to degrade MC-LR. After 72 h of GF3 cultivation, the MC-LR content was reduced by 34.1% and 26.7% at initial 0.1 μg/mL and 0.45 μg/mL concentrations, respectively. The high stress resistance of the GF3 to toxic MC-LR is provided by a 1.5-fold activation of catalase activity and a change in the reduced glutathione content. Additionally, during the MC-LR biotransformation, a MC-LR-GSH conjugate and linearized MC-LR were identified. The linearized MC-LR in the presence of fungi capable of degrading MCs was revealed for the first time. Using Daphnia magna as a bioindicator, it was shown that the MC-LR biotransformation led to the formation of less toxic intermediates. The toxicity of the fungal filtrate is reduced by five times compared to the abiotic control. Our findings enhance the understanding of the role that ascomycete fungi have as potential bioagents for cyanoHABs to control and detoxify water bodies. Full article

(1) Background: The first-line treatment for patients with focal or segmental dystonia with a craniocervical distribution is still the intramuscular injection of botulinum neurotoxin (BoNT). However, some patients experience primary or secondary treatment failure from this potential immunogenic therapy. Deep brain stimulation (DBS) may then be used as a backup strategy in this situation. (2) Methods: Here, we reviewed the current study literature to answer a specific question regarding the efficacy and safety of the use of DBS, particularly for cervical dystonia (CD) and Meige syndrome (MS) in patients with documented treatment failure under BoNT. (3) Results: There are only two studies with the highest level of evidence in this area. Despite this clear limitation, in the context of the narrowly defined research question of this paper, it is possible to report 161 patients with CD or MS who were included in studies that were able to show a statistically significant reduction in dystonic symptoms using DBS. Safety and tolerability data appeared adequate. However, much of the information is based on retrospective observations. (4) Conclusions: The evidence base in this area is in need of further scientific investigation. Most importantly, more randomized, controlled and double-blind trials are needed, possibly including a head-to-head comparison of DBS and BoNT. Full article

Mycotoxins are abiotic hazards whose contamination occurs at the pre- and post-harvest stages of the maize value chain, with animal exposure through contaminated feed leading to their excretion into milk. Currently, only aflatoxin M1 is regulated in milk products. Since feed materials and complete feed present a multi-mycotoxin composition and are the main mycotoxin source into milk, it is important to recognize the occurrence of multiple toxins and their co-occurrence in this highly consumed food product. The aim of this study was to determine the content of regulated and emerging mycotoxins in milk samples, which allowed for evaluating the occurrence and co-occurrence patterns of different mycotoxins known to contaminate feed materials and complete animal feed. Human exposure considering the occurrence patterns obtained was also estimated. Aflatoxins, fumonisins, zearalenone, and emerging mycotoxins were among the mycotoxins found to be present in the 100 samples analyzed. Concentrations ranged from 0.006 to 16.3 μg L⁻¹, with no sample exceeding the AFM1 maximum level. Though several mycotoxins were detected, no exceeding values were observed considering the TDI or PMTDI. It can be concluded that the observed exposure does not pose a health risk to milk consumers, though it is important to recognize vulnerable age groups. Full article

An adsorbent material derived from alfalfa leaves was prepared and further characterized, and its efficacy for removing aflatoxin B₁ (AFB₁) was investigated. Characterization consisted of the use of attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), environmental scanning electron microscopy (ESEM), X-ray fluorescence spectroscopy (XRF), X-ray diffraction (XRD), point of zero charge (pH_pzc), zeta potential (ζ-potential), UV-Vis diffuse reflectance spectroscopy, and spectral analysis. To determine the adsorption capacity against AFB₁ (250 ng AFB₁/mL), pH-dependent and avian intestinal in vitro models were used. The adsorbent inclusion percentage was 0.5% (w/w). In general, the pH-dependent model gave adsorption percentages of 98.2%, 99.9%, and 98.2%, evaluated at pH values of 2, 5, and 7, respectively. However, when the avian intestinal model was used, it was observed that the adsorption percentage of AFB₁ significantly decreased (88.8%). Based on the characterization results, it is proposed that electrostatic, non-electrostatic, and the formation of chlorophyll-AFB₁ complexes were the main mechanisms for AFB₁ adsorption. From these results, it can be concluded that the adsorbent derived from alfalfa leaves could be used as an effective material for removing AFB₁ in in vitro digestion models that mimic the physiological reality. Full article

Pectoralis Minor Syndrome (PMS) causes significant discomfort due to the compression of the neurovascular bundle within the retropectoralis minor space. Botulinum neurotoxin (BoNT) injections have emerged as a potential treatment method; however, their effectiveness depends on accurately locating the injection site. In this study, we aimed to identify optimal BoNT injection sites for PMS treatment. We used twenty-nine embalmed and eight non-embalmed human cadavers to determine the origin and intramuscular arborization of the pectoralis minor muscle (Pm) via manual dissection and Sihler’s nerve staining techniques. Our findings showed the Pm’s origin near an oblique line through the suprasternal notch, with most neural arborization within the proximal three-fourths of the Pm. Blind dye injections validated these results, effectively targeting the primary neural arborized area of the Pm at the oblique line’s intersection with the second and third ribs. We propose BoNT injections at the arborized region within the Pm’s proximal three-fourths, or the C region, for PMS treatment. These findings guide clinicians towards safer, more effective BoNT injections. Full article

Genetically modified MON 89034 corn (Zea mays L.) expressing Bacillus thuringiensis (Bt) insecticidal proteins, viz. Cry1A.105 and Cry2Ab2, is a biotechnological option being considered for the management of the major corn pest in Indonesia, the Asian corn borer (Ostrinia furnacalis (Guenée) (Lepidoptera: Crambidae)). As a part of a proactive resistance-management program for MON 89034 corn in Indonesia, we assessed the baseline susceptibility of field-collected populations of O. furnacalis to Cry1A.105 and Cry2Ab2 proteins. Dose–response bioassays using the diet-dipping method indicated that the lethal concentration (LC₅₀) values of Cry1A.105 and Cry2Ab2 in 24 different field populations of O. furnacalis ranged from 0.006 to 0.401 µg/mL and from 0.044 to 4.490 µg/mL, respectively, while the LC₉₅ values ranged from 0.069 to 15.233 µg/mL for Cry1A.105 and from 3.320 to 277.584 µg/mL for Cry2Ab2. The relative resistance ratios comparing the most tolerant field populations and an unselected laboratory population were 6.0 for Cry1A.105 and 2.0 for Cry2Ab2 based on their LC₅₀ values. Some field populations were more susceptible to both proteins than the unselected laboratory population. The LC₉₉ and its 95% fiducial limits across the field populations were calculated and proposed as candidate diagnostic concentrations. These data provide a basis for resistance monitoring in Bt Corn and further support building resistance-management strategies in Indonesia. Full article

Background: Plant-derived pyrrolizidine alkaloids (PAs) in feed cause metabolic disturbances in farm animals resulting in high economic losses worldwide. The molecular pathways affected by these PAs in cells and tissues are not yet fully understood. The objective of the study was to examine the dose-dependent effects of orally applied PAs derived from tansy ragwort in midlactation dairy cows. Methods: Twenty Holstein dairy cows were treated with target exposures of 0, 0.47, 0.95 and 1.91 mg of total PA/kg of body weight/d in control, PA1, PA2 and PA3, respectively, for 28 days. Liver tissue biopsy and plasma and milk samples were taken at day 28 of treatment to assess changes in metabolic pathways. A targeted metabolomics approach was performed to detect the metabolite profiles in all compartments. Results: The PA-affected metabolite profiling in liver tissue, plasma and milk revealed changes in three substrate classes: acylcarnitines (ACs), phosphatidylcholines (PCs) and sphingomyelins (SMs). In addition, in the plasma, amino acid concentrations were affected by PA exposure. Conclusions: PA exposure disturbed liver metabolism at many sites, especially devastating pathways related to energy metabolism and to amino acid utilization, most likely based on mitochondrial oxidative stress. The effects on the milk metabolite profile may have consequences for milk quality. Full article

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

The climate is changing in Europe: average temperatures are increasing, and so is the frequency of extreme weather events. Climate change has a severe impact on areas suitable for growing certain crops and on food safety, for example, affecting the occurrence of the aflatoxin contamination of maize. The aim of this study was to obtain insights into the impact of climate change on possible changes in land use in Europe, particularly in areas suitable for maize cultivation, and on the probability of the mycotoxin contamination of maize in order to give directions for long-term adaptation to climate change. By combining a land use model and a mycotoxin prediction model, the suitability of land for maize cultivation and the probability of aflatoxin contamination were estimated for suitable areas in Europe, comparing the current climate with the 2050 scenario. In 2050, the occurrence of aflatoxin contamination in Europe is predicted to severely increase, especially in Central and Southern Europe. More northern regions, presently unsuitable for maize cultivation, will become suitable for maize cultivation in 2050. In the baseline scenario, most regions suitable for maize cultivation have a low probability of aflatoxin contamination, whereas in 2050, about half of the regions suitable for maize cultivation have a medium to high probability of aflatoxin contamination. Regions for safely growing maize for human consumption will shift from the southern to the northern half of Europe. Full article

A field biologist was bitten by a female Nikolsky’s viper (Vipera berus nikolskii) in Kharkiv Oblast, Ukraine. Two months later, the patient began to experience cold-induced vasospasm of the affected digit diagnosed as acquired Raynaud phenomenon. The patient had more than 30 occurrences during the single winter following the bite, but the signs and symptoms of Raynaud phenomenon disappeared with the end of winter. This report describes the case and puts it into context with the literature on the topic of toxin-induced peripheral vasospastic disorders and their potential importance in snakebite envenoming. Full article

This study aimed to evaluate the efficacy of botulinum toxin type A (BoNT/A) in patients with temporomandibular disorders (TMDs) associated with masticatory muscle pain (MMP) and headaches. This randomized, double-blind, placebo-controlled pilot study is the first clinical trial to evaluate both disorders simultaneously. Twenty-one patients with myogenous TMD were randomly assigned to two groups. The experimental and control groups received injections of either BoNT/A or saline into the sites showing tenderness after palpation of a total of 16 muscle areas, including each masseter, a temporalis, splenius capitis, sternocleidomastoid, and trapezius muscle. During each visit, the clinical effects, based on the intensity of orofacial pain (OVAS), headache (HVAS), number of tender points (TPs), maximum mouth opening (MMO), and headache frequency (HF), were evaluated at four time points, namely, pre-injection and 4, 8, and 12 weeks after the injection, in both groups. Friedman and Mann–Whitney tests were used for the analyses. In the experimental group, the reductions in OVAS, TP, HVAS, and HF showed significant differences over time, excluding MMO, whereas there was no significant difference in any of the variables in the control group. In addition, the decline in TPs was significantly different between the experimental and control groups at all time points, especially after 4 and 12 weeks, compared to that during pre-injection. In conclusion, treatment with BoNT/A was relatively effective for masticatory muscle pain caused by TMDs and headache compared to the saline placebo. Full article

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

Calamus tenuis is a shrub species distributed across South Asia. It grows well in diversified habitats and tends to dominate plants in the surrounding environment. The phytotoxicity of C. tenuis and the action of its phytochemicals against other plant species could explain its dominant behavior. Compounds with phytotoxic activity are in high demand as prospective sources of ecofriendly bioherbicides. Therefore, we investigated the phytotoxicity of C. tenuis. Aqueous methanol extracts of this plant species significantly limited the growth of four test plant species, two monocots (barnyard grass and timothy), and two dicots (alfalfa and cress), in a dose- and species-dependent manner. Bio-directed chromatographic isolation of the C. tenuis extracts yielded two major active substances: a novel compound, calamulactone {(S)-methyl 8-(5-oxo-2,5-dihydrofuran-2-yl) octanoate}, and 3-oxo-α-ionone. Both of the identified compounds exerted strong growth inhibitory effects on cress and timothy seedlings. The concentrations of 3-oxo-α-ionone and calamulactone required to limit the growth of the cress seedlings by 50% (I₅₀) were 281.6–199.5 and 141.1–105.5 µM, respectively, indicating that the effect of calamulactone was stronger with lower I₅₀ values. Similarly, the seedlings of timothy also showed a considerably higher sensitivity to calamulactone (I₅₀: 40.5–84.4 µM) than to 3-oxo-α-ionone (I₅₀: 107.8–144.7 µM). The findings indicated that the leaves of C. tenuis have marked growth-inhibitory potential, and could affect surrounding plants to exert dominance over the surrounding plant community. Moreover, the two identified phytotoxic substances might play a key role in the phytotoxicity of C. tenuis, and could be a template for bioherbicide development. This paper was the first to report calamulactone and its phytotoxicity. Full article

The effects of deoxynivalenol (DON, 50 µg/mL) on the zebrafish liver and intestine were studied. Differentially expressed genes (DEGs) from mRNA and lncRNA were analyzed by RNA seq. Gene Ontology (GO) and signaling pathways were studied where the top 30 DEGs of each type of RNA were involved. The results showed there were 2325 up-regulated and 934 down-regulated DEGs of lncRNA in the intestinal tract, and 95 up-regulated genes and 211 down-regulated genes in the liver, respectively. GO functional annotation analysis showed that lncRNA was enriched in the biological processes, involving the RNA splicing, CSF1-CSF1R complexes, and MAP kinase activity. DEGs of lncRNA located in the KEGG signal pathways include the C-type lectin receptor signaling and the NOD-like receptor signaling pathways. Metabolism involves the biosynthesis of indole alkaloids, cancer pathways for human disease, MAPK and Rap1signaling pathways for environmental information processing, necroptosis and focal adhesion for cell processes. The mRNA gene expression analysis showed there were 1939 up-regulated, 1172 down-regulated genes and 866 up-regulated, 1211 down-regulated genes in the intestine and liver of zebrafish, respectively. This study provides transcriptome analysis and toxicological investigation of DON in the zebrafish liver and intestine, offering insights into gene expression patterns and potential detoxification pathways. Full article

Streptococcus pneumoniae is the leading cause of community-acquired pneumonia. The pore-forming cholesterol-dependent cytolysin (CDC) pneumolysin (PLY) and the physiological metabolite hydrogen peroxide (H₂O₂) can greatly increase the virulence of pneumococci. Although most studies have focused on the contribution of both virulence factors to the course of pneumococcal infection, it is unknown whether or how H₂O₂ can affect PLY activity. Of note, S. pneumoniae exploits endogenous H₂O₂ as an intracellular signalling molecule to modulate the activity of several proteins. Here, we demonstrate that H₂O₂ negatively affects the haemolytic activity of PLY in a concentration-dependent manner. Prevention of cysteine-dependent sulfenylation upon substitution of the unique and highly conserved cysteine residue to serine in PLY significantly reduces the toxin’s susceptibility to H₂O₂ treatment and completely abolishes the ability of DTT to activate PLY. We also detect a clear gradual correlation between endogenous H₂O₂ generation and PLY release, with decreased H₂O₂ production causing a decline in the release of PLY. Comparative transcriptome sequencing analysis of the wild-type S. pneumoniae strain and three mutants impaired in H₂O₂ production indicates enhanced expression of several genes involved in peptidoglycan (PG) synthesis and in the production of choline-binding proteins (CPBs). One explanation for the impact of H₂O₂ on PLY release is the observed upregulation of the PG bridge formation alanyltransferases MurM and MurN, which evidentially negatively affect the PLY release. Our findings shed light on the significance of endogenous pneumococcal H₂O₂ in controlling PLY activity and release. Full article

Background: Repetitive intramuscular injections of botulinum neurotoxin type A (BoNT/A) are the treatment of choice in patients with cervical dystonia (CD). As soon as BoNT therapy is initiated, the natural course of CD cannot be observed anymore. Nevertheless, the present study focuses on the “presumed” course of disease severity under the assumption that no BoNT therapy had been performed. The “experienced” benefit is compared with the “presumed” worsening. Methods: Twenty-seven BoNT/A long-term-treated CD patients were recruited. They had to assess the remaining severity of CD in percent of its severity at the start of BoNT therapy (RS-%). Then, they had to draw the course of severity from the onset of symptoms to the start of BoNT/A therapy (CoDB graph), as well as the course of severity from the start of BoNT/A therapy until the day of recruitment (CoDA graph). Then, they were instructed to presume the development of CD severity from the day of the start of BoNT/A therapy until the day of recruitment under the assumption that no BoNT/A therapy had been performed, and to assess the maximal severity they could presume in percent of the severity at the start of BoNT therapy (IS-%). Then, they had to draw the “presumed” development of CD severity (CoDI graph). The “experienced” change in disease severity and the “presumed” change since the start of BoNT/A therapy were compared and correlated with a variety of demographical and treatment-related data, including the actual severity of CD at the day of recruitment, which was assessed using the TSUI score and the actual dose per session (ADOSE). Results: No CD patients expected an improvement without BoNT therapy. “Presumed” worsening ((IS-%)-100) was about 50% in the mean and did not correlate with the “experienced” benefit (100-(RS-%)). However, IS-% was significantly correlated with ATSUI and ADOSE. Conclusion: Obviously, CD patients have the opinion that their CD would have further progressed and worsened if no BoNT/A therapy had been performed. Thus, the total benefit of BoNT/A therapy for a patient with CD is a combination of the “experienced” benefit under BoNT/A therapy and the prevented worsening of CD that the patient expects to occur without BoNT/A therapy. Full article