Toxins

Contamination of barley by deoxynivalenol (DON), a mycotoxin produced by Fusarium graminearum, causes considerable financial loss to the grain and malting industries. In this study, two atmospheric cold plasma (ACP) reactors were used to produce plasma-activated water (PAW) bubbles. The potential of PAW bubbles for the steeping of naturally infected barley (NIB) during the malting process was investigated. The PAW bubbles produced by treating water for 30 min using a bubble spark discharge (BSD) at low temperature resulted in the greatest concentration of oxygen-nitrogen reactive species (RONS). This treatment resulted in 57.3% DON degradation compared with 36.9% in the control sample; however, the same treatment reduced germination significantly (p < 0.05). Direct BSD ACP treatment for 20 min at low temperature and indirect treatment for 30 min increased the percentage of germinated rootlets of the seedlings compared with the control. Considering both the DON reduction and germination improvement of barley seeds, continuous jet ACP treatment for 30 min performed better than the other treatments used in this study. At higher temperature of PAW bubbles, the concentration of RONS was significantly (p < 0.05) reduced. Based on quantitative polymerase chain reaction (qPCR) analysis and fungal culture tests, the PAW bubble treatment did not significantly reduce infection of NIB. Nonetheless, this study provides useful information for the malting industry for PAW treatment optimization and its use in barley steeping for DON reduction and germination improvement. Full article

Botulinum neurotoxin (BoNT), produced by Clostridium botulinum, cleaves proteins involved in neurotransmitter release, thereby triggering flaccid paralyses, which are responsible for botulism. BoNT is classified into seven serotypes (BoNT/A-G); BoNT/A and BoNT/B are used as medical therapeutics and anti-wrinkle reagents. In this study, we investigated the efficacy of BoNT/CD, a mosaic toxin of BoNT/C and BoNT/D, to assess its potential as a therapeutic alternative for BoNT/A. In a cultured neuron assay, BoNT/CD cleaved syntaxin and SNAP-25 with higher efficacy than BoNT/C and BoNT/A. Intramuscularly administrated BoNT/CD induced dose-dependent muscle paralysis, and the paralysis lasted ~21 days in a mouse digit abduction score assay (BoNT/A-induced paralysis lasted ~30 days). BoNT/C failed to induce local paralysis without systemic toxicity. Multiple alignment analyses of the amino acid sequences of the receptor binding domain (H_C) of eight BoNT/CDs and two BoNT/Ds showed sequence clustering in five groups. Comparing BoNT/CD strain 003-9 (BoNT/CD_003-9) and strain 6813 (BoNT/CD₆₈₁₃) showed that both BoNT/CDs displayed similar efficacies in cultured neurons, but BoNT/CD_003-9 displayed higher efficacy in a mouse model than BoNT/CD₆₈₁₃. These findings suggest that BoNT/CD may be a potential alternative for patients who do not respond to existing BoNT-based therapeutics. Full article

This study was conducted to determine if a low monotonic dose of zearalenone (ZEN) affects the immunohistochemical expression (IE) of oestrogen receptor alpha (ERα) and oestrogen receptor beta (ERβ) in the intestines of sexually immature gilts. Group C (control group; n = 18) gilts were given a placebo. Group E (experimental group; n = 18) gilts were dosed orally with 40 μg ZEN /kg body weight (BW), each day before morning feeding. Samples of intestinal tissue were collected post-mortem six times. The samples were stained to analyse the IE of ERα and Erβ in the scanned slides. The strongest response was observed in ERα in the duodenum (90.387—average % of cells with ERα expression) and in ERβ in the descending colon (84.329—average % of cells with ERβ expression); the opposite response was recorded in the caecum (2.484—average % of cells with ERα expression) and the ascending colon (2.448—average % of cells with ERα expression); on the first two dates of exposure, the digestive tract had to adapt to ZEN in feed. The results of this study, supported by a mechanistic interpretation of previous research findings, suggest that ZEN performs numerous functions in the digestive tract. Full article

Background: Intravesical injection of Botulinum toxin A (BoNT-A) and platelet-rich plasma (PRP) have been reported to alleviate bladder pain and decrease nocturia in patients with refractory interstitial cystitis/bladder pain syndrome (IC/BPS). Both treatments are novel and there has no comparison between them. This study compared the therapeutic effects and adverse events between IC/BPS patients receiving PRP or BoNT-A injections. Materials and Methods: This study retrospectively analyzed female patients with IC/BPS who were refractory to conventional treatment and received BoNT-A (n = 26) or PRP (n = 30) injections within the previous two years. Patients were arbitrarily treated with four monthly injections of PRP or a single injection of 100 U of BoNT-A. All injections were followed by cystoscopic hydrodistention. The primary endpoint was the global response assessment (GRA), and secondary endpoints were changes in the O’Leary-Sant IC symptom score, visual analog score (VAS) of bladder pain, voiding diary, and uroflow measures from baseline to six months after the first injection day. Results: The baseline demographics revealed no significant difference between groups. The GRA at one, three, and six months was similar between groups. A significant improvement in IC symptom scores was noted in both groups. Although VAS was significantly improved in overall patients, no significant difference was noted between the PRP and BoNT-A groups at 6 months. Only half of the study cohort had a GRA ≥2 at six months. An increase in the post-void residual was noted one month after the BoNT-A injection, but there was no difference between groups at three and six months. More patients reported dysuria (19.2% vs. 3.3%, p = 0.086) and urinary tract infection (UTI, 15.4% vs. 0%, p = 0.041) after BoNT-A injection than after the PRP injections. The time from the first injection to receiving alternative treatment was similar between groups. Conclusion: Both intravesical PRP and BoNT-A injections have similar efficacy in IC symptom improvement. However, only half of the study cohort had a GRA of ≥2 at the six-month follow-up BoNT-A injection carries a potential risk of UTI after treatment. Full article

Pig health is impaired and growth performance is reduced when exposed to deoxynivalenol (DON). The measurement of DON in individual feedstuffs and complete swine diets is variable because of the inconsistent distribution of mycotoxins in feed and the difficulties in obtaining representative samples. We investigated whether measuring DON and its metabolites in biological samples could be used as a predictor of DON ingestion by pigs. Blood samples were collected between 3 and 4 h after the morning meal and urine samples were quantitatively collected over a 24 h period on d 40 and 82 of the study to evaluate serum and urinary content of DON and DON metabolites (iso-deoxynivalenol, DON-3-glucuronide, DON-15-glcurunide, deepoxy-deoxynivalenol, iso-deepoxy-deoxynivalenol, deepoxy-deoxynivalenol-3-glucuronide, and deepoxy-deoxynivalenol-15-glucuronide). The intake of DON was positively correlated with urinary DON output. Similarly, there was an increase in serum DON level with increasing DON intake. Overall, it was found that DON intake correlated with DON concentration in urine and blood serum when samples were collected under controlled conditions. Analyzing DON levels in urine and blood serum could be used to predict a pig’s DON intake. Full article

Frequent outbreaks of harmful cyanobacterial blooms and the cyanotoxins they produce not only seriously jeopardize the health of freshwater ecosystems but also directly affect the survival of aquatic organisms. In this study, the dynamic characteristics and response patterns of transcriptomes and gut microbiomes in gastropod Bellamya aeruginosa were investigated to explore the underlying response mechanisms to toxic cyanobacterial exposure. The results showed that toxic cyanobacteria exposure induced overall hepatopancreatic transcriptome changes. A total of 2128 differentially expressed genes were identified at different exposure stages, which were mainly related to antioxidation, immunity, and metabolism of energy substances. In the early phase (the first 7 days of exposure), the immune system may notably be the primary means of resistance to toxin stress, and it performs apoptosis to kill damaged cells. In the later phase (the last 7 days of exposure), oxidative stress and the degradation activities of exogenous substances play a dominant role, and nutrient substance metabolism provides energy to the body throughout the process. Microbiomic analysis showed that toxic cyanobacteria increased the diversity of gut microbiota, enhanced interactions between gut microbiota, and altered microbiota function. In addition, the changes in gut microbiota were correlated with the expression levels of antioxidant-, immune-, metabolic-related differentially expressed genes. These results provide a comprehensive understanding of gastropods and intestinal microbiota response to toxic cyanobacterial stress. Full article

Aristolochic acid (AA) is notorious for inducing nephrotoxicity, but the influence of sex on AA-induced kidney injury was not clear. This study sought to investigate sex differences in kidney dysfunction and tubular injury induced by AA. Male and female mice were bilaterally orchiectomized and ovariectomized, respectively. Fourteen days after gonadectomy, the mice were intraperitoneally injected with AA (10 mg/kg body weight/day) daily for 2 days and sacrificed 7 days after the first injection. Body weight, kidney function, and tubular structure were assessed. When compared between male and female non-gonadectomized mice, AA-induced body weight loss was greater in male mice than in female mice. Functional and structural damages in male kidneys were markedly induced by AA injection, but kidneys in AA-injected female mice showed no or mild damages. Ovariectomy had no effect on AA-induced nephrotoxic acute kidney injury in female mice. However, orchiectomy significantly reduced body weight loss, kidney dysfunction, and tubular injury in AA-induced nephrotoxicity in male mice. This study has demonstrated that testis causes AA-induced nephrotoxic acute kidney injury. Full article

In this study, a simple and efficient magnetic solid-phase extraction (MSPE) strategy was developed to simultaneously purify and enrich nine mycotoxins in fruits, with the magnetic covalent organic framework nanomaterial Fe₃O₄@COF(TAPT–DHTA) as an adsorbent. The Fe₃O₄@COF(TAPT–DHTA) was prepared by a simple template precipitation polymerization method, using Fe₃O₄ as magnetic core, and 1,3,5-tris-(4-aminophenyl) triazine (TAPT) and 2,5-dihydroxy terephthalaldehyde (DHTA) as two building units. Fe₃O₄@COF(TAPT–DHTA) could effectively capture the targeted mycotoxins by virtue of its abundant hydroxyl groups and aromatic rings. Several key parameters affecting the performance of the MSPE method were studied, including the adsorption solution, adsorption time, elution solvent, volume and time, and the amount of Fe₃O₄@COF(TAPT–DHTA) nanomaterial. Under optimized MSPE conditions, followed by analysis with UHPLC–MS/MS, a wide linear range (0.05–200 μg kg⁻¹), low limits of detection (0.01–0.5 μg kg⁻¹) and satisfactory recovery (74.25–111.75%) were achieved for the nine targeted mycotoxins. The established method was further successfully validated in different kinds of fruit samples. Full article

Patients with chronic kidney disease (CKD) have a higher cardiovascular risk compared to the average population, and this is partially due to the plasma accumulation of solutes known as uremic toxins. The binding of some solutes to plasma proteins complicates their removal via conventional therapies, e.g., hemodialysis. Protein-bound uremic toxins originate either from endogenous production, diet, microbial metabolism, or the environment. Although the impact of diet on uremic toxicity in CKD is difficult to quantify, nutrient intake plays an important role. Indeed, most uremic toxins are gut-derived compounds. They include Maillard reaction products, hippurates, indoles, phenols, and polyamines, among others. In this review, we summarize the findings concerning foods and dietary components as sources of uremic toxins or their precursors. We then discuss their endogenous metabolism via human enzyme reactions or gut microbial fermentation. Lastly, we present potential dietary strategies found to be efficacious or promising in lowering uremic toxins plasma levels. Aligned with current nutritional guidelines for CKD, a low-protein diet with increased fiber consumption and limited processed foods seems to be an effective treatment against uremic toxins accumulation. Full article

During the past decades, the gut microbiome emerged as a key player in kidney disease. Dysbiosis-related uremic toxins together with pro-inflammatory mediators are the main factors in a deteriorating kidney function. The toxicity of uremic compounds has been well-documented in a plethora of pathophysiological mechanisms in kidney disease, such as cardiovascular injury (CVI), metabolic dysfunction, and inflammation. Accumulating data on the detrimental effect of uremic solutes in kidney disease supported the development of many strategies to restore eubiosis. Fecal microbiota transplantation (FMT) spread as an encouraging treatment for different dysbiosis-associated disorders. In this scenario, flourishing studies indicate that fecal transplantation could represent a novel treatment to reduce the uremic toxins accumulation. Here, we present the state-of-the-art concerning the application of FMT on kidney disease to restore eubiosis and reverse the retention of uremic toxins. Full article

Receptor binding is a prerequisite process to exert the mosquitocidal activity of the Cry4Ba toxin of Bacillus thuringiensis subsp. israelensis. The beta-sheet prism (domain II) and beta-sheet sandwich (domain III) of the Cry4Ba toxin have been implicated in receptor binding, albeit the precise binding mechanisms of these remain unclear. In this work, alanine scanning was used to determine the contribution to receptor binding of some aromatic and hydrophobic residues on the surface of domains II and III that are predicted to be responsible for binding to the Aedes aegypti membrane-bound alkaline phosphatase (Aa-mALP) receptor. Larvicidal activity assays against A. aegypti larvae revealed that aromatic residues (Trp₃₂₇ on the β2 strand, Tyr₃₄₇ on the β3–β4 loop, and Tyr₃₅₉ on the β4 strand) of domain II were important to the toxicity of the Cry4Ba toxin. Quantitative binding assays using enzyme-linked immunosorbent assay (ELISA) showed similar decreasing trends in binding to the Aa-mALP receptor and in toxicity of the Cry4Ba mutants Trp327Ala, Tyr347Ala, and Tyr359Ala, suggesting that a possible function of these surface-exposed aromatic residues is receptor binding. In addition, binding assays of the Cry4Ba toxin to the mutants of the binding residues Gly₅₁₃, Ser₄₉₀, and Phe₄₉₇ of the Aa-mALP receptor supported the binding function of Trp₃₂₇, Tyr₃₄₇, and Tyr₃₅₉ of the Cry4Ba toxin, respectively. Altogether, our results showed for the first time that aromatic residues on a side surface of the Cry4Ba domain II function in receptor binding. This finding provides greater insight into the possible molecular mechanisms of the Cry4Ba toxin. Full article

Staphylococcal enterotoxin A (SEA) has presented enormous difficulties in dairy food safety and the sensitive detection of SEA provides opportunities for effective food safety controls and staphylococcal food poisoning tracebacks. Herein, a novel aggregation-induced emission (AIE)-based sandwich lateral flow immunoassay (LFIA) was introduced to detect SEA by using red-emissive AIE nanoparticles (AIENPs) as the fluorescent nanoprobe. The nanoprobe was constructed by directly immobilising antibodies on boronate-tagged AIENPs (PBA-AIENPs) via a boronate affinity reaction, which exhibited a high SEA-specific affinity and remarkable fluorescent performance. Under optimal conditions, the ultrasensitive detection of SEA in pasteurised milk was achieved within 20 min with a limit of detection of 0.04 ng mL⁻¹. The average recoveries of the PBA-AIENP-LFIA ranged from 91.3% to 117.6% and the coefficient of variation was below 15%. It was also demonstrated that the PBA-AIENP-LFIA had an excellent selectivity against other SE serotypes. Taking advantage of the excellent sensitivity of this approach, real chicken and salad samples were further analysed, with a high versatility and accuracy. The proposed PBA-AIENP-LFIA platform shows promise as a potent tool for the identification of additional compounds in food samples as well as an ideal test method for on-site detections. Full article

Venom expressed by the nearly 50,000 species of spiders on Earth largely remains an untapped reservoir of a diverse array of biomolecules with potential for pharmacological and agricultural applications. A large fraction of the noxious components of spider venoms are a functionally diverse family of structurally related polypeptides with an inhibitor cystine knot (ICK) motif. The cysteine-rich nature of these toxins makes structural elucidation difficult, and most studies have focused on venom components from the small handful of medically relevant spider species such as the highly aggressive Brazilian wandering spider Phoneutria nigriventer. To alleviate difficulties associated with the study of ICK toxins in spiders, we devised a comprehensive approach to explore the evolutionary patterns that have shaped ICK functional diversification using venom gland transcriptomes and proteomes from phylogenetically distinct lineages of wandering spiders and their close relatives. We identified 626 unique ICK toxins belonging to seven topological elaborations. Phylogenetic tests of episodic diversification revealed distinct regions between cysteine residues that demonstrated differential evidence of positive or negative selection, which may have structural implications towards the specificity and efficacy of these toxins. Increased taxon sampling and whole genome sequencing will provide invaluable insights to further understand the evolutionary processes that have given rise to this diverse class of toxins. Full article

Diseases contribute to attainment of less than 50% of the local groundnut potential yield in Kenya. This study aimed to evaluate the agronomic characteristics (flowering and germination), disease incidence, yield performance (biomass, harvest index, 100-pod, 100-seed, and total pod weight), and aflatoxin accumulation in six peanut varieties. A field experiment was conducted using four newly improved peanut varieties: CG9, CG7, CG12, and ICGV-SM 90704 (Nsinjiro), and two locally used varieties: Homabay local (control) and 12991, and in a randomized complete block design with three replications. The disease identification followed the International Crop Research Institute for the Semi-Arid Tropics (ICRISAT) rating scale and further isolation of fungal contaminants was conducted by a direct plating technique using potato dextrose agar. The aflatoxin levels in the peanuts were determined after harvesting using the ultrahigh performance liquid chromatography and fluorescence detection (UHPLC-FLD) technique. ICGV-SM 90704 showed the least average disease incidence of 1.31 ± 1.75%, (P < 0.05); the lowest total aflatoxin levels (1.82 ± 1.41 μg kg⁻¹) with a range 0.00–0.85 μg kg⁻¹ for total aflatoxins and a range 0.00–1.24 μg kg⁻¹ for Aflatoxin B₁. The locally used varieties (12991 and the control) revealed the highest disease incidence (5.41 ± 8.31% and 7.41 ± 1.88%), respectively. ICGV-SM 90704 was the best performing among all the six varieties with an average total pod weight (9.22 ± 1.19 kg), 100-pod weight (262.93 ± 10.8 g), and biomass of (27.21 ± 5.05 kg) per row. The 12991 variety and the control showed the least total pod weight (1.60 ± 0.28 and 1.50 ± 1.11 kg, respectively) (P = 0.0001). The newly improved varieties showed lower disease rates, low levels of aflatoxins, and higher yields than the locally used varieties. Full article

End-stage renal disease (ESRD) patients rely on renal replacement therapies to survive. Hemodialysis (HD), the most widely applied treatment, is responsible for the removal of excess fluid and uremic toxins (UTs) from blood, particularly those with low molecular weight (MW < 500 Da). The development of high-flux membranes and more efficient treatment modes, such as hemodiafiltration, have resulted in improved removal rates of UTs in the middle molecular weight range. However, the concentrations of protein-bound uremic toxins (PBUTs) remain essentially untouched. Due to the high binding affinity to large proteins, such as albumin, PBUTs form large complexes (MW > 66 kDa) which are not removed during HD and their accumulation has been strongly associated with the increased morbidity and mortality of patients with ESRD. In this review, we describe adsorption- and displacement-based approaches currently being studied to enhance the removal of PBUTs. The development of mixed matrix membranes (MMMs) with selective adsorption properties, infusion of compounds capable of displacing UTs from their binding site on albumin, and competitive binding membranes show promising results, but the road to clinical application is still long, and further investigation is required. Full article

Brown spider envenomation results in dermonecrosis, characterized by an intense inflammatory reaction. The principal toxins of brown spider venoms are phospholipase-D isoforms, which interact with different cellular membrane components, degrade phospholipids, and generate bioactive mediators leading to harmful effects. The Loxosceles intermedia phospholipase D, LiRecDT1, possesses a loop that modulates the accessibility to the active site and plays a crucial role in substrate. In vitro and in silico analyses were performed to determine aspects of this enzyme’s substrate preference. Sphingomyelin d18:1/6:0 was the preferred substrate of LiRecDT1 compared to other Sphingomyelins. Lysophosphatidylcholine 16:0/0:0 was preferred among other lysophosphatidylcholines, but much less than Sphingomyelin d18:1/6:0. In contrast, phosphatidylcholine d18:1/16:0 was not cleaved. Thus, the number of carbon atoms in the substrate plays a vital role in determining the optimal activity of this phospholipase-D. The presence of an amide group at C2 plays a key role in recognition and activity. In silico analyses indicated that a subsite containing the aromatic residues Y228 and W230 appears essential for choline recognition by cation-π interactions. These findings may help to explain why different cells, with different phospholipid fatty acid compositions exhibit distinct susceptibilities to brown spider venoms. Full article

Spinal cord injuries (SCI) have a profound impact on autonomic systems, sometimes resulting in multi-organ dysfunction, including of the neurogenic bladder. Autonomic dysreflexia (AD) is commonly seen in patients with SCI above T6 when the injured cord develops a deregulated sympathetic reflex, which can be induced by bladder sensation and can cause hypertensive crisis. While intravesical injection of botulinum toxin A (Botox) is a standard therapy for neurogenic detrusor overactivity, the role of Botox for AD has rarely been described. This study reviewed the medical records of SCI patients who reported AD and received either detrusor or urethral sphincter injection with Botox. The primary endpoint is the subjective improvement of AD. The secondary endpoint is a change in videourodynamic parameters before and after Botox injection. A total of 200 patients were enrolled for analysis. There were 125 (62.5%) patients in the detrusor injection group, and 75 (37.5%) in the urethral sphincter injection group. There were 79 (63.2%) patients in the detrusor injection group and 43 (57.3%) in the urethral sphincter injection group reporting moderate or marked improvement. Detrusor injection leads to a greater improvement in AD, probably because of decreased detrusor pressure and increased compliance after Botox injection. Urethral sphincter injection appears to have a modest effect on AD, despite general improvements in the voiding parameters of videourodynamic study. Full article

The existence of aflatoxin M₁ (AFM₁) in raw milk results in economic losses and public health risks. This research aims to examine the capability of bentonite to adsorb and/or eliminate AFM₁ from various raw milk types. In addition, the effects of numerous bentonites (HAFR 1, 2, 3 and 4) on the nutritional characteristics of the milk were studied. Our findings revealed that goat milk had the highest value of AFM₁ (490.30 ng/L) in comparison to other milks. AFM₁ adsorption was influenced by applying bentonite (0.5 and 1 g) in a concentration-dependent manner for different time intervals (from 0 to 12 h). The percentage of AFM₁ reached the maximum adsorption level after 12 h to 100, 98.5 and 98% for bentonites HAFR 3, 1 and 2, respectively. HAFR 3 (1 g bentonite) presented higher adsorption efficiency than other bentonites used in the phosphate buffer saline (PBS) and milk. Residual levels of AFM₁ reached their lowest values of 0 and 1.5 ng/L while using HAFR 3 in PBS and milk, respectively. With regard to the influence of bentonite on the nutritional characteristics of milk, there was an increase in fat, protein and solid non-fat ratio while using HAFR 3 and 4, yet decreased lactose in comparison with the control. Scanning Electron Microscopy and Fourier Transform-Infrared Spectroscopy both identified bentonites as superior AFM₁ binders. The results demonstrated that bentonite, particularly HAFR 3, was the most effective adsorbent and could thus be a promising candidate for the decontamination of AFM₁ in milk. Full article

Ants are one of the important groups of venomous animals with about 14,000 described species. Studies so far focused on the discovery of venom proteins are only available for limited stinging ants, and the proteinaceous compositions of the stingless ants are completely unknown. Here, we used the transcriptomic approach to identify venom components from the yellow meadow ant, Lasius flavus, a stingless ant. The transcriptomic analysis yielded an extraordinary simplicity of the venom expression profile, with 17 venom proteins, such as phospholipase B, odorant binding protein, and apolipoprotein D. Ten of them were discovered as novel toxins for future functional investigations. Quantitative real time PCR analysis revealed that genes encoding the identified venom proteins display exclusively or highly expression profiles in venom glands, validating them as venom compositions. Our findings contribute to the understanding of the evolutional diversity of toxins between stinging and stingless ants. Full article