Toxins

14 pages, 4846 KiB

Open AccessArticle

The Putative Histone Methyltransferase DOT1 Regulates Aflatoxin and Pathogenicity Attributes in Aspergillus flavus

by Linlin Liang, Yinghang Liu, Kunlong Yang, Guinan Lin, Zhangling Xu, Huahui Lan, Xiuna Wang and Shihua Wang

Toxins 2017, 9(7), 232; https://doi.org/10.3390/toxins9070232 - 24 Jul 2017

Cited by 25 | Viewed by 6196

Abstract

Lysine methyltransferases transfer methyl groups in specific lysine sites, which regulates a variety of important biological processes in eukaryotes. In this study, we characterized a novel homolog of the yeast methyltransferase DOT1 in A. flavus, and observed the roles of dot1 [...] Read more.

Lysine methyltransferases transfer methyl groups in specific lysine sites, which regulates a variety of important biological processes in eukaryotes. In this study, we characterized a novel homolog of the yeast methyltransferase DOT1 in A. flavus, and observed the roles of dot1 in A. flavus. Deletion of dot1 showed a significant decrease in conidiation, but an increase in sclerotia formation. A change in viability to multiple stresses was also found in the Δdot1 mutant. Additionally, aflatoxin (AF) production was found severely impaired in the Δdot1 mutant. Further analysis by qRT-PCR revealed that the transcription of AF structural genes and their regulator gene aflS were prominently suppressed in the Δdot1 mutant. Furthermore, our data revealed that Dot1 is important for colonizing maize seeds in A. flavus. Our research indicates that Dot1 is involved in fungal development, aflatoxin biosynthesis and fungal virulence in A. flavus, which might provide a potential target for controlling A. flavus with new strategies. Full article

(This article belongs to the Collection Aflatoxins)

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21 pages, 3476 KiB

Open AccessArticle

Microcystin Prevalence throughout Lentic Waterbodies in Coastal Southern California

by Meredith D. A. Howard, Carey Nagoda, Raphael M. Kudela, Kendra Hayashi, Avery Tatters, David A. Caron, Lilian Busse, Jeff Brown, Martha Sutula and Eric D. Stein

Toxins 2017, 9(7), 231; https://doi.org/10.3390/toxins9070231 - 22 Jul 2017

Cited by 38 | Viewed by 6883

Abstract

Toxin producing cyanobacterial blooms have increased globally in recent decades in both frequency and intensity. Despite the recognition of this growing risk, the extent and magnitude of cyanobacterial blooms and cyanotoxin prevalence is poorly characterized in the heavily populated region of southern California. [...] Read more.

Toxin producing cyanobacterial blooms have increased globally in recent decades in both frequency and intensity. Despite the recognition of this growing risk, the extent and magnitude of cyanobacterial blooms and cyanotoxin prevalence is poorly characterized in the heavily populated region of southern California. Recent assessments of lentic waterbodies (depressional wetlands, lakes, reservoirs and coastal lagoons) determined the prevalence of microcystins and, in some cases, additional cyanotoxins. Microcystins were present in all waterbody types surveyed although toxin concentrations were generally low across most habitats, as only a small number of sites exceeded California’s recreational health thresholds for acute toxicity. Results from passive samplers (Solid Phase Adsorption Toxin Tracking (SPATT)) indicated microcystins were prevalent throughout lentic waterbodies and that traditional discrete samples underestimated the presence of microcystins. Multiple cyanotoxins were detected simultaneously in some systems, indicating multiple stressors, the risk of which is uncertain since health thresholds are based on exposures to single toxins. Anatoxin-a was detected for the first time from lakes in southern California. The persistence of detectable microcystins across years and seasons indicates a low-level, chronic risk through both direct and indirect exposure. The influence of toxic cyanobacterial blooms is a more complex stressor than presently recognized and should be included in water quality monitoring programs. Full article

(This article belongs to the Collection Freshwater HABs and Health in a Changing World)

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14 pages, 1488 KiB

Open AccessArticle

SiMa Cells for a Serotype Specific and Sensitive Cell-Based Neutralization Test for Botulinum Toxin A and E

by Nicola Bak, Shalini Rajagopal, Paul Stickings and Dorothea Sesardic

Toxins 2017, 9(7), 230; https://doi.org/10.3390/toxins9070230 - 20 Jul 2017

Cited by 8 | Viewed by 8500

Abstract

Botulinum toxins (BoNTs), of which there are seven serotypes, are among the most potent neurotoxins, with serotypes A, B and E causing human botulism. Antitoxins form the first line of treatment for botulism, and functional, highly sensitive in vitro methods for toxin neutralization [...] Read more.

Botulinum toxins (BoNTs), of which there are seven serotypes, are among the most potent neurotoxins, with serotypes A, B and E causing human botulism. Antitoxins form the first line of treatment for botulism, and functional, highly sensitive in vitro methods for toxin neutralization are needed to replace the current in vivo methods used for determination of antitoxin potency. In this preliminary proof of concept study, we report the development of a neutralization test using the neuroblastoma SiMa cell line. The assay is serotype specific for either BoNT/A or BoNT/E, which both cleave unique sequences on SNAP-25 within SiMa cells. The end point is simple immunodetection of cleaved SNAP-25 from cell lysates with antibodies detecting only the newly exposed sequence on SNAP-25. Neutralizing antibodies prevent the toxin-induced cleavage of SNAP-25. The toxin neutralization assay, with an EC50 of ~2 mIU/mL determined with a standardized reference antiserum, is more sensitive than the mouse bioassays. Relevance was demonstrated with commercial and experimental antitoxins targeting different functional domains, and of known in vivo neutralizing activities. This is the first report describing a simple, specific, in vitro cell-based assay for the detection of neutralizing antibodies against BoNT/A and BoNT/E with a sensitivity exceeding that of the mouse bioassay. Full article

(This article belongs to the Special Issue Botulinum Neurotoxins Antibody and Vaccine)

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15 pages, 8662 KiB

Open AccessArticle

The Uremic Toxin Indoxyl Sulfate Accelerates Thrombotic Response after Vascular Injury in Animal Models

by Malgorzata Karbowska, Tomasz W. Kaminski, Natalia Marcinczyk, Tomasz Misztal, Tomasz Rusak, Lukasz Smyk and Dariusz Pawlak

Toxins 2017, 9(7), 229; https://doi.org/10.3390/toxins9070229 - 19 Jul 2017

Cited by 35 | Viewed by 6327

Abstract

Chronic kidney disease (CKD) patients are at high risk for thrombotic events. Indoxyl sulfate (IS) is one of the most potent uremic toxins that accumulates during CKD. Even though IS is associated with an increased risk for cardiovascular disease, its impact on thrombotic [...] Read more.

Chronic kidney disease (CKD) patients are at high risk for thrombotic events. Indoxyl sulfate (IS) is one of the most potent uremic toxins that accumulates during CKD. Even though IS is associated with an increased risk for cardiovascular disease, its impact on thrombotic events still remains not fully understood. The purpose of the study was to evaluate the direct effect of IS on thrombotic process. We examined the impact of acute exposure to IS on thrombus development induced by electric current in Wistar rats, intravital thrombus formation after laser-induced injury in the mice endothelium, coagulation profile, clot formation dynamics, platelet aggregations, and erythrocyte osmotic resistance. IS doses: 10, 30 and 100 mg/kg body weight (b.w.) increased weight of thrombus induced by electric current in dose-dependent manner (p < 0.001). Furthermore, two highest IS doses increased laser-induced thrombus formation observed via confocal system (increase in fluorescence intensity and total thrombus area (p < 0.01)). Only the highest IS dose decreased clotting time (p < 0.01) and increased maximum clot firmness (p < 0.05). IS did not affect blood morphology parameters and erythrocyte osmotic resistance, but augmented collagen-induced aggregation. Obtained data indicate that IS creates prothrombotic state and contributes to more stable thrombus formation. Thus, we concluded that IS may be one of crucial uremic factors promoting thrombotic events in CKD patients. Full article

(This article belongs to the Special Issue Novel Issues in Uremic Toxicity)

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30 pages, 948 KiB

Open AccessReview

Emerging Fusarium and Alternaria Mycotoxins: Occurrence, Toxicity and Toxicokinetics

by Sophie Fraeyman, Siska Croubels, Mathias Devreese and Gunther Antonissen

Toxins 2017, 9(7), 228; https://doi.org/10.3390/toxins9070228 - 18 Jul 2017

Cited by 249 | Viewed by 13612

Abstract

Emerging Fusarium and Alternaria mycotoxins gain more and more interest due to their frequent contamination of food and feed, although in vivo toxicity and toxicokinetic data are limited. Whereas the Fusarium mycotoxins beauvericin, moniliformin and enniatins particularly contaminate grain and grain-based products, Alternaria [...] Read more.

Emerging Fusarium and Alternaria mycotoxins gain more and more interest due to their frequent contamination of food and feed, although in vivo toxicity and toxicokinetic data are limited. Whereas the Fusarium mycotoxins beauvericin, moniliformin and enniatins particularly contaminate grain and grain-based products, Alternaria mycotoxins are also detected in fruits, vegetables and wines. Although contamination levels are usually low (µg/kg range), higher contamination levels of enniatins and tenuazonic acid may occasionally occur. In vitro studies suggest genotoxic effects of enniatins A, A1 and B1, beauvericin, moniliformin, alternariol, alternariol monomethyl ether, altertoxins and stemphyltoxin-III. Furthermore, in vitro studies suggest immunomodulating effects of most emerging toxins and a reproductive health hazard of alternariol, beauvericin and enniatin B. More in vivo toxicity data on the individual and combined effects of these contaminants on reproductive and immune system in both humans and animals is needed to update the risk evaluation by the European Food Safety Authority. Taking into account new occurrence data for tenuazonic acid, the complete oral bioavailability, the low total body clearance in pigs and broiler chickens and the limited toxicity data, a health risk cannot be completely excluded. Besides, some less known Alternaria toxins, especially the genotoxic altertoxins and stemphyltoxin III, should be incorporated in risk evaluation as well. Full article

(This article belongs to the Collection Leading Opinions (Closed))

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19 pages, 3559 KiB

Open AccessArticle

A Bacterial Toxin with Analgesic Properties: Hyperpolarization of DRG Neurons by Mycolactone

by Ok-Ryul Song, Han-Byul Kim, Samuel Jouny, Isabelle Ricard, Alexandre Vandeputte, Nathalie Deboosere, Estelle Marion, Christophe J. Queval, Pierre Lesport, Emmanuel Bourinet, Daniel Henrion, Seog Bae Oh, Guillaume Lebon, Guillaume Sandoz, Edouard Yeramian, Laurent Marsollier and Priscille Brodin

Toxins 2017, 9(7), 227; https://doi.org/10.3390/toxins9070227 - 18 Jul 2017

Cited by 29 | Viewed by 6839

Abstract

Mycolactone, a polyketide molecule produced by Mycobacterium ulcerans, is the etiological agent of Buruli ulcer. This lipid toxin is endowed with pleiotropic effects, presents cytotoxic effects at high doses, and notably plays a pivotal role in host response upon colonization by the [...] Read more.

Mycolactone, a polyketide molecule produced by Mycobacterium ulcerans, is the etiological agent of Buruli ulcer. This lipid toxin is endowed with pleiotropic effects, presents cytotoxic effects at high doses, and notably plays a pivotal role in host response upon colonization by the bacillus. Most remarkably, mycolactone displays intriguing analgesic capabilities: the toxin suppresses or alleviates the pain of the skin lesions it inflicts. We demonstrated that the analgesic capability of mycolactone was not attributable to nerve damage, but instead resulted from the triggering of a cellular pathway targeting AT₂ receptors (angiotensin II type 2 receptors; AT₂R), and leading to potassium-dependent hyperpolarization. This demonstration paves the way to new nature-inspired analgesic protocols. In this direction, we assess here the hyperpolarizing properties of mycolactone on nociceptive neurons. We developed a dedicated medium-throughput assay based on membrane potential changes, and visualized by confocal microscopy of bis-oxonol-loaded Dorsal Root Ganglion (DRG) neurons. We demonstrate that mycolactone at non-cytotoxic doses triggers the hyperpolarization of DRG neurons through AT₂R, with this action being not affected by known ligands of AT₂R. This result points towards novel AT₂R-dependent signaling pathways in DRG neurons underlying the analgesic effect of mycolactone, with the perspective for the development of new types of nature-inspired analgesics. Full article

(This article belongs to the Section Bacterial Toxins)

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14 pages, 5431 KiB

Open AccessArticle

Ouabain Protects Human Renal Cells against the Cytotoxic Effects of Shiga Toxin Type 2 and Subtilase Cytotoxin

by María M. Amaral, Magalí C. Girard, Romina S. Álvarez, Adrienne W. Paton, James C. Paton, Horacio A. Repetto, Flavia Sacerdoti and Cristina A. Ibarra

Toxins 2017, 9(7), 226; https://doi.org/10.3390/toxins9070226 - 18 Jul 2017

Cited by 6 | Viewed by 5231

Abstract

Hemolytic uremic syndrome (HUS) is one of the most common causes of acute renal failure in children. The majority of cases are associated with Shiga toxin (Stx)-producing Escherichia coli (STEC). In Argentina, HUS is endemic and presents the highest incidence rate in the [...] Read more.

Hemolytic uremic syndrome (HUS) is one of the most common causes of acute renal failure in children. The majority of cases are associated with Shiga toxin (Stx)-producing Escherichia coli (STEC). In Argentina, HUS is endemic and presents the highest incidence rate in the world. STEC strains expressing Stx type 2 (Stx2) are responsible for the most severe cases of this pathology. Subtilase cytotoxin (SubAB) is another STEC virulence factor that may contribute to HUS pathogenesis. To date, neither a licensed vaccine nor effective therapy for HUS is available for humans. Considering that Ouabain (OUA) may prevent the apoptosis process, in this study we evaluated if OUA is able to avoid the damage caused by Stx2 and SubAB on human glomerular endothelial cells (HGEC) and the human proximal tubule epithelial cell (HK-2) line. HGEC and HK-2 were pretreated with OUA and then incubated with the toxins. OUA protected the HGEC viability from Stx2 and SubAB cytotoxic effects, and also prevented the HK-2 viability from Stx2 effects. The protective action of OUA on HGEC and HK-2 was associated with a decrease in apoptosis and an increase in cell proliferation. Our data provide evidence that OUA could be considered as a therapeutic strategy to avoid the renal damage that precedes HUS. Full article

(This article belongs to the Collection Shiga Toxins)

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15 pages, 2776 KiB

Open AccessArticle

Characterization of Enterotoxigenic Bacillus cereus sensu lato and Staphylococcus aureus Isolates and Associated Enterotoxin Production Dynamics in Milk or Meat-Based Broth

by Laura Walker-York-Moore, Sean C. Moore and Edward M. Fox

Toxins 2017, 9(7), 225; https://doi.org/10.3390/toxins9070225 - 15 Jul 2017

Cited by 20 | Viewed by 8092

Abstract

Bacillus cereus sensu lato species, as well as Staphylococcus aureus, are important pathogenic bacteria which can cause foodborne illness through the production of enterotoxins. This study characterised enterotoxin genes of these species and examined growth and enterotoxin production dynamics of isolates when [...] Read more.

Bacillus cereus sensu lato species, as well as Staphylococcus aureus, are important pathogenic bacteria which can cause foodborne illness through the production of enterotoxins. This study characterised enterotoxin genes of these species and examined growth and enterotoxin production dynamics of isolates when grown in milk or meat-based broth. All B. cereus s. l. isolates harboured nheA, hblA and entFM toxin genes, with lower prevalence of bceT and hlyII. When grown at 16 °C, toxin production by individual B. cereus s. l. isolates varied depending on the food matrix; toxin was detected at cell densities below 5 log₁₀(CFU/mL). At 16 °C no staphylococcal enterotoxin C (SEC) production was detected by S. aureus isolates, although low levels of SED production was noted. At 30 °C all S. aureus isolates produced detectable enterotoxin in the simulated meat matrix, whereas SEC production was significantly reduced in milk. Relative to B. cereus s. l. toxin production, S. aureus typically required reaching higher cell numbers to produce detectable levels of enterotoxin. Phylogenetic analysis of the sec and sel genes suggested population evolution which correlated with animal host adaptation, with subgroups of bovine isolates or caprine/ovine isolates noted, which were distinct from human isolates. Taken together, this study highlights the marked differences in the production of enterotoxins both associated with different growth matrices themselves, but also in the behaviour of individual strains when exposed to different food matrices. Full article

(This article belongs to the Section Bacterial Toxins)

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11 pages, 6443 KiB

Open AccessArticle

Differences in Genetic Background Contribute to Pseudomonas Exotoxin A-Induced Hepatotoxicity in Rats

by Chien-Chao Chiu, Yu-Chih Wang, Wen-Ching Huang, Yi-Hsun Chen, Shao-Wen Hung, Yen-Te Huang, Hsiao-Li Chuang and Yi-Chih Chang

Toxins 2017, 9(7), 224; https://doi.org/10.3390/toxins9070224 - 15 Jul 2017

Cited by 5 | Viewed by 4284

Abstract

Pseudomonas aeruginosa exotoxin A (PEA) causes severe hepatotoxicity in experimental animals and is useful in investigations of immune-mediated liver injury. However, strain differences in the sensitivity to PEA-induced hepatotoxicity in rats remains be elucidated. In this study, we determined the severity of PEA-induced [...] Read more.

Pseudomonas aeruginosa exotoxin A (PEA) causes severe hepatotoxicity in experimental animals and is useful in investigations of immune-mediated liver injury. However, strain differences in the sensitivity to PEA-induced hepatotoxicity in rats remains be elucidated. In this study, we determined the severity of PEA-induced hepatotoxicity in six genetically different rat strains. Male LE (Long Evans), Wistar, F344, WKY, BN/SsN and LEW rats were administered a single intravenous injection of PEA (20 μg/kg). Significantly elevated serum ALT and AST levels, massive necrosis and hemorrhage, and numerous TUNEL-positive hepatocytes were observed in BN/SsN rats. In contrast, low levels of ALT and AST as well as mild changes in liver histopathology were observed in Wistar and F344 rats. Moderate levels of hepatic injuries were observed in LE, WKY, and LEW rats. Pro-inflammatory cytokines including TNF-α, IL-2 and IL-6 serum levels were markedly increased in BN/SsN rats compared to Wistar and F344 rats. However, the hepatic levels of low density lipoprotein receptor-related protein (LRP), which functions as the PEA receptor, were not significantly different in each strain. Taken together, we suggest that BN/SsN is the most sensitive rat strain, whereas Wistar and F344 were the most resistant rat strains to PEA-induced liver damage. The different genetic background of rat strains plays an important role in the susceptibility to PEA-induced epatotoxicity that may depend on immune-regulation but not LRP receptor levels. Full article

(This article belongs to the Section Bacterial Toxins)

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15 pages, 2174 KiB

Open AccessArticle

The Occurrence of Zearalenone in South Korean Feedstuffs between 2009 and 2016

by Hansub Chang, Woori Kim, Ju-Hee Park, Dongho Kim, Choong-Ryeol Kim, Soohyun Chung and Chan Lee

Toxins 2017, 9(7), 223; https://doi.org/10.3390/toxins9070223 - 15 Jul 2017

Cited by 55 | Viewed by 6091

Abstract

Mycotoxins produced by Fusarium plant pathogen species have harmful effects on humans and livestock by natural contamination in food and feed. Zearalenone, one of the well-known Fusarium mycotoxins, causes hyperestrogenism and toxicosis resulting in reproductive dysfunction in animals. This study investigated the occurrence [...] Read more.

Mycotoxins produced by Fusarium plant pathogen species have harmful effects on humans and livestock by natural contamination in food and feed. Zearalenone, one of the well-known Fusarium mycotoxins, causes hyperestrogenism and toxicosis resulting in reproductive dysfunction in animals. This study investigated the occurrence of zearalenone in feedstuffs (compound feeds, feed ingredients) between 2009 and 2016 in South Korea to obtain information on zearalenone contamination in feeds for management. A total of 653 animal feed samples (494 compound feeds, 159 feed ingredients) produced domestically were sampled five times from 2009 to 2016 (2009, 2010, 2012, 2014, and 2016) from feed factories in South Korea. The levels of zearalenone were analyzed every year by high-performance liquid chromatography (HPLC) after pretreatment with an immunoaffinity column showing limit of detection (LOD) and limit of quantification (LOQ) of 0.1–3 μg/kg and 0.3–8 μg/kg, respectively. Four feed samples out of 494 compound feeds exceeded the EU and South Korea commission regulations over the eight-year test period, and no feed ingredients exceeded the guidelines. Full article

(This article belongs to the Collection Fusarium Toxins – Relevance for Human and Animal Health)

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11 pages, 372 KiB

Open AccessArticle

Ability of Soil Isolated Actinobacterial Strains to Prevent, Bind and Biodegrade Ochratoxin A

by Rachelle El Khoury, Florence Mathieu, Ali Atoui, Hiba Kawtharani, Anthony El Khoury, Charbel Afif, Richard G. Maroun and André El Khoury

Toxins 2017, 9(7), 222; https://doi.org/10.3390/toxins9070222 - 14 Jul 2017

Cited by 23 | Viewed by 4800

Abstract

Ochratoxin A (OTA) is one of the most important mycotoxins, and contaminates several agricultural products, particularly cereals, grapes, maize, barley, spices and coffee. The aim of this project was to reduce the levels of OTA by supplementing the artificially contaminated solutions with seven [...] Read more.

Ochratoxin A (OTA) is one of the most important mycotoxins, and contaminates several agricultural products, particularly cereals, grapes, maize, barley, spices and coffee. The aim of this project was to reduce the levels of OTA by supplementing the artificially contaminated solutions with seven strains of actinobacteria (AT10, AT8, SN7, MS1, ML5, G10 and PT1) in order to evaluate their capacity for binding and metabolizing the OTA, as well as their ability to reduce the expression of the genes responsible for its production in A. carbonarius. In the first part of this study, we evaluated the capacity of Streptomyces strains for binding OTA on their surfaces after 0, 30 and 60 min of incubation with PBS solution supplemented with OTA. In the second part, we tested the ability of these strains, as well as their supernatants, to detoxify the ISP2 medium. Finally, we studied the effect of the Streptomyces cocultured with Aspergillus carbonarius on the expression of OTA biosynthesis genes. Results showed that, among the strains co-cultured with A. carbonarius, the strain G10 was able to reduce the expression of acpks, acOTApks, acOTAnrps and vea genes, thus reducing OTA from solid PDA medium to 13.50% of reduction. This strain was remarkably able to detoxify and bind OTA up to 47.07%. Strain AT8 was stronger in detoxifying OTA (52.61%), but had no significant effect on the studied gene expression. Full article

(This article belongs to the Special Issue 1^st International MYCOKEY Conference: Advances on Mycotoxin Reduction in the Food and Feed Chain)

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15 pages, 2995 KiB

Open AccessArticle

Phage Display Analysis of Monoclonal Antibody Binding to Anthrax Toxin Lethal Factor

by Jason M. Goldstein, Joo Lee, Xiaoling Tang, Anne E. Boyer, John R. Barr, Dennis A. Bagarozzi Jr. and Conrad P. Quinn

Toxins 2017, 9(7), 221; https://doi.org/10.3390/toxins9070221 - 13 Jul 2017

Cited by 5 | Viewed by 6580

Abstract

AVR1674 and AVR1675 are monoclonal antibodies (mAbs) that bind with high specificity to anthrax toxin lethal factor (LF) and lethal toxin (LTx). These mAbs have been used as pivotal reagents to develop anthrax toxin detection tests using mass spectrometry. The mAbs were demonstrated [...] Read more.

AVR1674 and AVR1675 are monoclonal antibodies (mAbs) that bind with high specificity to anthrax toxin lethal factor (LF) and lethal toxin (LTx). These mAbs have been used as pivotal reagents to develop anthrax toxin detection tests using mass spectrometry. The mAbs were demonstrated to bind LF with good affinity (K_D 10⁻⁷–10⁻⁹ M) and to enhance LF-mediated cleavage of synthetic peptide substrates in vitro. Sequence analysis indicated that the mAbs shared 100% amino acid identity in their complementarity determining regions (CDR). A phage display library based on a combinatorial library of random heptapeptides fused to the pIII coat protein of M13 phage was enriched and screened to identify peptide sequences with mAb binding properties. Selection and sequence analysis of 18 anti-LF-reactive phage clones identified a 7-residue (P1–P7) AVR1674/1675 consensus target binding sequence of T_P1-X_P2-K/R_P3-D_P4-D/E_P5-Z_P6-X/Z_P7 (X = aromatic, Z = non-polar). The phage peptide sequence with highest affinity binding to AVR1674/1675 was identified as T-F-K-D-E-I-V. Synthetic oligopeptides were designed based on the phage sequences and interacted with mAbs with high affinity (K_D~ 10⁻⁹ M). Single amino acid substitutions of A, H, or Q in the peptides identified positions P1–P5 as critical residues for mAb-peptide interactions. CLUSTALW alignment of phage sequences with native LF implicated residues 644–650 (sequence T-H-Q-D-E-I-Y) as a putative linear epitope component located within a structural loop (L2) of LF Domain IV. The activation effects of these mAbs contribute to the analytic sensitivity of function-based LF detection assays. Full article

(This article belongs to the Collection Anthrax Toxins)

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12 pages, 1230 KiB

Open AccessArticle

Content of the Saponin Protodioscin in Brachiaria spp. from the Eastern Plains of Colombia

by Maria C. Lozano, Nhora M. Martinez and Gonzalo J. Diaz

Toxins 2017, 9(7), 220; https://doi.org/10.3390/toxins9070220 - 13 Jul 2017

Cited by 16 | Viewed by 5448

Abstract

Protodioscin is used as a marker of saponin content that could cause hepatotoxicity in ruminants. In Brachiaria spp. from two regions of the Colombian Eastern Plains (east mountain range of the Andean—“piedemonte” and Ariari River Valley) were determined this metabolite at 14 and [...] Read more.

Protodioscin is used as a marker of saponin content that could cause hepatotoxicity in ruminants. In Brachiaria spp. from two regions of the Colombian Eastern Plains (east mountain range of the Andean—“piedemonte” and Ariari River Valley) were determined this metabolite at 14 and 28 days post-cutting under different climatic conditions. No protodioscin was detected in B. dictyoneura or B. humidicola. In B. brizantha, B. decumbens and B. ruziziensis x B. decumbens x B. brizantha (hybrid), protodioscin content corresponded to an interaction between species, post-cutting time and season. Concentrations ≥1% (minimum toxic level) were recorded in B. decumbens and the hybrid, and to a lesser extent in B. brizantha. The concentration of protodioscin was higher at 28 days, when the pastures are suitable for consumption. B. brizantha accumulated the lowest saponin concentration, whereas the hybrid had the highest levels, particularly in the “piedemonte” and during drought (3.37%). Dry season favored the protodioscin concentration in B. decumbens (in river valley) and in the hybrid (in “piedemonte”). In the latter, there was a positive correlation with temperature and a negative with humidity, which are typical characteristics of dry periods. This is the first report of protodioscin content in the hybrid. Full article

(This article belongs to the Section Plant Toxins)

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10 pages, 1651 KiB

Open AccessArticle

The Mode of Action of Cyclo(l-Ala-l-Pro) in Inhibiting Aflatoxin Production of Aspergillus flavus

by Kurin Iimura, Tomohiro Furukawa, Toshiyoshi Yamamoto, Lumi Negishi, Michio Suzuki and Shohei Sakuda

Toxins 2017, 9(7), 219; https://doi.org/10.3390/toxins9070219 - 12 Jul 2017

Cited by 16 | Viewed by 5463

Abstract

Cyclo(l-Ala-l-Pro) inhibits aflatoxin production in aflatoxigenic fungi without affecting fungal growth. The mode of action of cyclo(l-Ala-l-Pro) in inhibiting aflatoxin production of Aspergillus flavus was investigated. A glutathione S-transferase (GST) of the fungus, designated [...] Read more.

Cyclo(l-Ala-l-Pro) inhibits aflatoxin production in aflatoxigenic fungi without affecting fungal growth. The mode of action of cyclo(l-Ala-l-Pro) in inhibiting aflatoxin production of Aspergillus flavus was investigated. A glutathione S-transferase (GST) of the fungus, designated AfGST, was identified as a binding protein of cyclo(l-Ala-l-Pro) in an experiment performed using cyclo(l-Ala-l-Pro)-immobilized Sepharose beads. Cyclo(l-Ala-l-Pro) specifically bound to recombinant AfGST and inhibited its GST activity. Ethacrynic acid, a known GST inhibitor, inhibited the GST activity of recombinant AfGST and aflatoxin production of the fungus. Ethacrynic acid reduced the expression level of AflR, a key regulatory protein for aflatoxin production, similar to cyclo(l-Ala-l-Pro). These results suggest that cyclo(l-Ala-l-Pro) inhibits aflatoxin production by affecting GST function in A. flavus, and that AfGST inhibitors are possible candidates as selective aflatoxin production inhibitors. Full article

(This article belongs to the Collection Aflatoxins)

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12 pages, 2287 KiB

Open AccessArticle

Genotypic Regulation of Aflatoxin Accumulation but Not Aspergillus Fungal Growth upon Post-Harvest Infection of Peanut (Arachis hypogaea L.) Seeds

by Walid Ahmed Korani, Ye Chu, Corley Holbrook, Josh Clevenger and Peggy Ozias-Akins

Toxins 2017, 9(7), 218; https://doi.org/10.3390/toxins9070218 - 12 Jul 2017

Cited by 16 | Viewed by 6182

Abstract

Aflatoxin contamination is a major economic and food safety concern for the peanut industry that largely could be mitigated by genetic resistance. To screen peanut for aflatoxin resistance, ten genotypes were infected with a green fluorescent protein (GFP)—expressing Aspergillus flavus strain. Percentages of [...] Read more.

Aflatoxin contamination is a major economic and food safety concern for the peanut industry that largely could be mitigated by genetic resistance. To screen peanut for aflatoxin resistance, ten genotypes were infected with a green fluorescent protein (GFP)—expressing Aspergillus flavus strain. Percentages of fungal infected area and fungal GFP signal intensity were documented by visual ratings every 8 h for 72 h after inoculation. Significant genotypic differences in fungal growth rates were documented by repeated measures and area under the disease progress curve (AUDPC) analyses. SICIA (Seed Infection Coverage and Intensity Analyzer), an image processing software, was developed to digitize fungal GFP signals. Data from SICIA image analysis confirmed visual rating results validating its utility for quantifying fungal growth. Among the tested peanut genotypes, NC 3033 and GT-C20 supported the lowest and highest fungal growth on the surface of peanut seeds, respectively. Although differential fungal growth was observed on the surface of peanut seeds, total fungal growth in the seeds was not significantly different across genotypes based on a fluorometric GFP assay. Significant differences in aflatoxin B levels were detected across peanut genotypes. ICG 1471 had the lowest aflatoxin level whereas Florida-07 had the highest. Two-year aflatoxin tests under simulated late-season drought also showed that ICG 1471 had reduced aflatoxin production under pre-harvest field conditions. These results suggest that all peanut genotypes support A. flavus fungal growth yet differentially influence aflatoxin production. Full article

(This article belongs to the Section Mycotoxins)

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9 pages, 832 KiB

Open AccessArticle

Rehabilitation plus OnabotulinumtoxinA Improves Motor Function over OnabotulinumtoxinA Alone in Post-Stroke Upper Limb Spasticity: A Single-Blind, Randomized Trial

by Deidre Devier, JoAnn Harnar, Leandro Lopez, Allison Brashear and Glenn Graham

Toxins 2017, 9(7), 216; https://doi.org/10.3390/toxins9070216 - 11 Jul 2017

Cited by 15 | Viewed by 5672

Abstract

Background: OnabotulinumtoxinA (BoNT-A) can temporarily decrease spasticity following stroke, but whether there is an associated improvement in upper limb function is less clear. This study measured the benefit of adding weekly rehabilitation to a background of BoNT-A treatments for chronic upper limb spasticity [...] Read more.

Background: OnabotulinumtoxinA (BoNT-A) can temporarily decrease spasticity following stroke, but whether there is an associated improvement in upper limb function is less clear. This study measured the benefit of adding weekly rehabilitation to a background of BoNT-A treatments for chronic upper limb spasticity following stroke. Methods: This was a multi-center clinical trial. Thirty-one patients with post-stroke upper limb spasticity were treated with BoNT-A. They were then randomly assigned to 24 weeks of weekly upper limb rehabilitation or no rehabilitation. They were injected up to two times, and followed for 24 weeks. The primary outcome was change in the Fugl–Meyer upper extremity score, which measures motor function, sensation, range of motion, coordination, and speed. Results: The ‘rehab’ group significantly improved on the Fugl–Meyer upper extremity score (Visit 1 = 60, Visit 5 = 67) while the ‘no rehab’ group did not improve (Visit 1 = 59, Visit 5 = 59; p = 0.006). This improvement was largely driven by the upper extremity “movement” subscale, which showed that the ‘rehab’ group was improving (Visit 1 = 33, Visit 5 = 37) while the ‘no rehab’ group remained virtually unchanged (Visit 1 = 34, Visit 5 = 33; p = 0.034). Conclusions: Following injection of BoNT-A, adding a program of rehabilitation improved motor recovery compared to an injected group with no rehabilitation. Full article

(This article belongs to the Section Bacterial Toxins)

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11 pages, 915 KiB

Open AccessArticle

Dynamic Fumonisin B₂ Production by Aspergillus niger Intented Used in Food Industry in China

by Xiaomin Han, Hongru Jiang, Jin Xu, Jing Zhang and Fengqin Li

Toxins 2017, 9(7), 217; https://doi.org/10.3390/toxins9070217 - 9 Jul 2017

Cited by 14 | Viewed by 6508

Abstract

There are a total of 30 strains including 27 strains of Aspergillus niger intended used in Chinese food industry, two strains used as control and one strain isolated from corn for fumonisin (FB) production on 3 media. It was found that FB₂ [...] Read more.

There are a total of 30 strains including 27 strains of Aspergillus niger intended used in Chinese food industry, two strains used as control and one strain isolated from corn for fumonisin (FB) production on 3 media. It was found that FB₂ production by A. niger was function-dependent and highly related to culture media, as well as incubation time. All strains studied were unable to produce FB₁ and FB₃. Almost all strains were found to produce FB₂ on corn, rice and wheat bran. Based on their intended use in the food industry, the higher level of FB₂ producers were strains used for saccharifying enzyme (n = 13) production, followed by organic acid (n = 6), tannase (n = 7) and β-galactosidase (n = 1) production, with the FB₂ mean level of 3553–10,270 μg/kg, 1059–12,036 μg/kg, 3–7 μg/kg and 2–4 μg/kg on corn, 5455–9241 μg/kg, 559–2190 μg/kg, 4–9 μg/kg and 6–10 μg/kg on rice, 5959–7709 μg/kg, 9491–17,339 μg/kg, 8–14 μg/kg and 120–222 μg/kg on wheat bran, respectively. Comparatively, strains of Fusarium verticillioide were capable of producing fumonins simultaneously with broader spectrum including FB₁, FB₂ and FB₃, but at a much lower level. In conclusion, it is necessary to evaluate FB₂ production by A. niger before intended use in the food processing industry. Full article

(This article belongs to the Collection Understanding Mycotoxin Occurrence in Food and Feed Chains)

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14 pages, 5645 KiB

Open AccessArticle

Evaluation of Cyanea capillata Sting Management Protocols Using Ex Vivo and In Vitro Envenomation Models

by Thomas K. Doyle, Jasmine L. Headlam, Christie L. Wilcox, Eoin MacLoughlin and Angel A. Yanagihara

Toxins 2017, 9(7), 215; https://doi.org/10.3390/toxins9070215 - 7 Jul 2017

Cited by 18 | Viewed by 21084

Abstract

Lion’s mane jellyfish (Cyanea capillata) stings cause severe pain and can lead to dangerous systemic effects, including Irukandji-like syndrome. As is the case for most cnidarian stings, recommended medical protocols in response to such stings lack rigorous scientific support. In this [...] Read more.

Lion’s mane jellyfish (Cyanea capillata) stings cause severe pain and can lead to dangerous systemic effects, including Irukandji-like syndrome. As is the case for most cnidarian stings, recommended medical protocols in response to such stings lack rigorous scientific support. In this study, we sought to evaluate potential first aid care protocols using previously described envenomation models that allow for direct measurements of venom activity. We found that seawater rinsing, the most commonly recommended method of tentacle removal for this species, induced significant increases in venom delivery, while rinsing with vinegar or Sting No More^® Spray did not. Post-sting temperature treatments affected sting severity, with 40 min of hot-pack treatment reducing lysis of sheep’s blood (in agar plates), a direct representation of venom load, by over 90%. Ice pack treatment had no effect on sting severity. These results indicate that sting management protocols for Cyanea need to be revised immediately to discontinue rinsing with seawater and include the use of heat treatment. Full article

(This article belongs to the Section Animal Venoms)

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10 pages, 1966 KiB

Open AccessArticle

Screening and Validation of Highly-Efficient Insecticidal Conotoxins from a Transcriptome-Based Dataset of Chinese Tubular Cone Snail

by Bingmiao Gao, Chao Peng, Bo Lin, Qin Chen, Junqing Zhang and Qiong Shi

Toxins 2017, 9(7), 214; https://doi.org/10.3390/toxins9070214 - 6 Jul 2017

Cited by 20 | Viewed by 6947

Abstract

Most previous studies have focused on analgesic and anti-cancer activities for the conotoxins identified from piscivorous and molluscivorous cone snails, but little attention has been devoted to insecticidal activity of conotoxins from the dominant vermivorous species. As a representative vermivorous cone snail, the [...] Read more.

Most previous studies have focused on analgesic and anti-cancer activities for the conotoxins identified from piscivorous and molluscivorous cone snails, but little attention has been devoted to insecticidal activity of conotoxins from the dominant vermivorous species. As a representative vermivorous cone snail, the Chinese tubular cone snail (Conus betulinus) is the dominant Conus species inhabiting the South China Sea. We sequenced related venom transcriptomes from C. betulinus using both the next-generation sequencing and traditional Sanger sequencing technologies, and a comprehensive library of 215 conotoxin transcripts was constructed. In our current study, six conotoxins with potential insecticidal activity were screened out from our conotoxin library by homologous search with a reported positive control (alpha-conotoxin ImI from C. imperialis) as the query. Subsequently, these conotoxins were synthesized by chemical solid-phase and oxidative folding for further insecticidal activity validation, such as MTT assay, insect bioassay and homology modeling. The final results proved insecticidal activities of our achieved six conotoxins from the transcriptome-based dataset. Interestingly, two of them presented a lot of high insecticidal activity, which supports their usefulness for a trial as insecticides in field investigations. In summary, our present work provides a good example for high throughput development of biological insecticides on basis of the accumulated genomic resources. Full article

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

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15 pages, 3135 KiB

Open AccessArticle

Whole Transcriptomic Analysis Provides Insights into Molecular Mechanisms for Toxin Biosynthesis in a Toxic Dinoflagellate Alexandrium catenella (ACHK-T)

by Yong Zhang, Shu-Fei Zhang, Lin Lin and Da-Zhi Wang

Toxins 2017, 9(7), 213; https://doi.org/10.3390/toxins9070213 - 5 Jul 2017

Cited by 38 | Viewed by 6513

Abstract

Paralytic shellfish toxins (PSTs), a group of neurotoxic alkaloids, are the most potent biotoxins for aquatic ecosystems and human health. Marine dinoflagellates and freshwater cyanobacteria are two producers of PSTs. The biosynthesis mechanism of PSTs has been well elucidated in cyanobacteria; however, it [...] Read more.

Paralytic shellfish toxins (PSTs), a group of neurotoxic alkaloids, are the most potent biotoxins for aquatic ecosystems and human health. Marine dinoflagellates and freshwater cyanobacteria are two producers of PSTs. The biosynthesis mechanism of PSTs has been well elucidated in cyanobacteria; however, it remains ambiguous in dinoflagellates. Here, we compared the transcriptome profiles of a toxin-producing dinoflagellate Alexandrium catenella (ACHK-T) at different toxin biosynthesis stages within the cell cycle using RNA-seq. The intracellular toxin content increased gradually in the middle G1 phase and rapidly in the late G1 phase, and then remained relatively stable in other phases. Samples from four toxin biosynthesis stages were selected for sequencing, and finally yielded 110,370 unigenes, of which 66,141 were successfully annotated in the known databases. An analysis of differentially expressed genes revealed that 2866 genes altered significantly and 297 were co-expressed throughout the four stages. These genes participated mainly in protein metabolism, carbohydrate metabolism, and the oxidation-reduction process. A total of 138 homologues of toxin genes were identified, but they altered insignificantly among different stages, indicating that toxin biosynthesis might be regulated translationally or post-translationally. Our results will serve as an important transcriptomic resource to characterize key molecular processes underlying dinoflagellate toxin biosynthesis. Full article

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

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16 pages, 2595 KiB

Open AccessArticle

Metabolism of the Marine Phycotoxin PTX-2 and Its Effects on Hepatic Xenobiotic Metabolism: Activation of Nuclear Receptors and Modulation of the Phase I Cytochrome P450

by Jimmy Alarcan, Estelle Dubreil, Antoine Huguet, Dominique Hurtaud-Pessel, Stefanie Hessel-Pras, Alfonso Lampen, Valérie Fessard and Ludovic Le Hegarat

Toxins 2017, 9(7), 212; https://doi.org/10.3390/toxins9070212 - 5 Jul 2017

Cited by 8 | Viewed by 5863

Abstract

PTX-2 is a marine biotoxin frequently found in shellfish that can lead to food intoxication in humans. Information regarding PTX-2 metabolism is scarce, and little is known of its effect on xenobiotic-metabolizing enzymes (XME) or its molecular pathways. The aim of this study [...] Read more.

PTX-2 is a marine biotoxin frequently found in shellfish that can lead to food intoxication in humans. Information regarding PTX-2 metabolism is scarce, and little is known of its effect on xenobiotic-metabolizing enzymes (XME) or its molecular pathways. The aim of this study was consequently to examine PTX-2 Phase I metabolism using rat and human liver S9 fractions, and also to assess the capability of PTX-2: (i) to modulate the gene expression of a panel of Phase I (CYP450) and II (UGT, SULT, NAT, and GST) enzymes, as well as the Phase III or 0 (ABC and SLCO) transporters in the human hepatic HepaRG cell line using qPCR; (ii) to induce specific CYP450 in HepaRG cells measured by immunolabeling detection and the measurement of the cells’ activities; and (iii) to activate nuclear receptors and induce CYP promoter activities in HEK-T and HepG2 transfected cell lines using transactivation and reporter gene assay, respectively. Our results indicate that PTX-2 hydroxylation occurred with both rat and human S9 fractions. Whereas PTX-2 mostly upregulated the gene expression of CYP1A1 and 1A2, no induction of these two CYP activities was observed. Lastly, PTX-2 did not act as an agonist of CAR or PXR. Due to its effects on some key XME, more attention should be paid to possible drug–drug interactions with phycotoxins, especially as shellfish can accumulate several phycotoxins as well as other kinds of contaminants. Full article

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

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13 pages, 1304 KiB

Open AccessArticle

Mechanisms for Differential Protein Production in Toxin–Antitoxin Systems

by Heather S. Deter, Roderick V. Jensen, William H. Mather and Nicholas C. Butzin

Toxins 2017, 9(7), 211; https://doi.org/10.3390/toxins9070211 - 4 Jul 2017

Cited by 26 | Viewed by 8077

Abstract

Toxin–antitoxin (TA) systems are key regulators of bacterial persistence, a multidrug-tolerant state found in bacterial species that is a major contributing factor to the growing human health crisis of antibiotic resistance. Type II TA systems consist of two proteins, a toxin and an [...] Read more.

Toxin–antitoxin (TA) systems are key regulators of bacterial persistence, a multidrug-tolerant state found in bacterial species that is a major contributing factor to the growing human health crisis of antibiotic resistance. Type II TA systems consist of two proteins, a toxin and an antitoxin; the toxin is neutralized when they form a complex. The ratio of antitoxin to toxin is significantly greater than 1.0 in the susceptible population (non-persister state), but this ratio is expected to become smaller during persistence. Analysis of multiple datasets (RNA-seq, ribosome profiling) and results from translation initiation rate calculators reveal multiple mechanisms that ensure a high antitoxin-to-toxin ratio in the non-persister state. The regulation mechanisms include both translational and transcriptional regulation. We classified E. coli type II TA systems into four distinct classes based on the mechanism of differential protein production between toxin and antitoxin. We find that the most common regulation mechanism is translational regulation. This classification scheme further refines our understanding of one of the fundamental mechanisms underlying bacterial persistence, especially regarding maintenance of the antitoxin-to-toxin ratio. Full article

(This article belongs to the Section Bacterial Toxins)

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22 pages, 5578 KiB

Open AccessArticle

Influence of Environmental Factors on the Production of Penitrems A–F by Penicillium crustosum

by Svetlana A. Kalinina, Annika Jagels, Benedikt Cramer, Rolf Geisen and Hans-Ulrich Humpf

Toxins 2017, 9(7), 210; https://doi.org/10.3390/toxins9070210 - 1 Jul 2017

Cited by 26 | Viewed by 6887

Abstract

Filamentous fungi produce a multitude of secondary metabolites, some of them known as mycotoxins, which are toxic to vertebrates and other animal groups in low concentrations. Among them, penitrems, which belong to the group of indole-diterpene mycotoxins, are synthesized by Penicillium and Aspergillus [...] Read more.

Filamentous fungi produce a multitude of secondary metabolites, some of them known as mycotoxins, which are toxic to vertebrates and other animal groups in low concentrations. Among them, penitrems, which belong to the group of indole-diterpene mycotoxins, are synthesized by Penicillium and Aspergillus genera and exhibit potent tremorgenic effects. This is the first complex study of the penitrems A–F production under the influence of different abiotic factors, e.g., media, incubation time, temperature, pH, light, water activity, and carbon and nitrogen source as well as oxidative and salt stress. For this purpose, penitrems A–F were isolated from Penicillium crustosum cultures and used as analytical standards. Among the carbon sources, glucose supplemented to the media at the concentration of 50 g/L, showed the strongest inducing effect on the biosynthesis of penitrems. Among nitrogen sources, glutamate was found to be the most favorable supplement, significantly increasing production of these secondary metabolites. CuSO4-promoted oxidative stress was also shown to remarkably stimulate biosynthesis of all penitrems. In contrast, the salt stress, caused by the elevated concentrations of NaCl, showed an inhibitory effect on the penitrem biosynthesis. Finally, cheese model medium elicited exceptionally high production of all members of the penitrems family. Obtained results give insides into the biosynthesis of toxicologically relevant penitrems A–F under different environmental factors and can be utilized to prevent food contamination. Full article

(This article belongs to the Section Mycotoxins)

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19 pages, 3922 KiB

Open AccessArticle

Probing the Characterization of the Interaction of Aflatoxins B1 and G1 with Calf Thymus DNA In Vitro

by Liang Ma, Jiaman Wang and Yuhao Zhang

Toxins 2017, 9(7), 209; https://doi.org/10.3390/toxins9070209 - 1 Jul 2017

Cited by 27 | Viewed by 8591

Abstract

The binding characterization of aflatoxins with calf thymus DNA (ctDNA) under physiological conditions was investigated. Multispectroscopic techniques, ctDNA melting, viscosity measurements, and molecular docking techniques were employed to elucidate the binding mechanism of the aflatoxins with DNA. The fluorescence results indicated that both [...] Read more.

The binding characterization of aflatoxins with calf thymus DNA (ctDNA) under physiological conditions was investigated. Multispectroscopic techniques, ctDNA melting, viscosity measurements, and molecular docking techniques were employed to elucidate the binding mechanism of the aflatoxins with DNA. The fluorescence results indicated that both aflatoxin B1 (AFB1) and aflatoxin G1 (AFG1) bound to the ctDNA, forming complexes through hydrogen bonding. The binding constants of AFB1 and AFG1 with ctDNA reached up to 10³ L·mol⁻¹ and 10⁴ L·mol⁻¹, respectively, and AFG1 exhibited a higher binding propensity than that of AFB1. Furthermore, both AFB1 and AFG1 bound to the ctDNA through groove binding, as evidenced by the results of the spectroscopic, iodide quenching effect, viscosity, and ctDNA melting measurements. Changes in the circular dichroism signal manifested that both AFB1 and AFG1 induced an increase in the right-handed helicity, but only minimally influenced the base stacking of the DNA. A molecular docking study of the aflatoxin’s binding with the DNA revealed a groove binding mode, which was driven mainly by hydrogen bonding. This study of aflatoxin–ctDNA interaction may provide novel insights into the toxicological effect of the mycotoxins. Full article

(This article belongs to the Collection Aflatoxins)

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10 pages, 4960 KiB

Open AccessArticle

Prenylflavonoid Isoxanthohumol Sensitizes MCF-7/ADR Cells to Doxorubicin Cytotoxicity via Acting as a Substrate of ABCB1

by Ming Liu, Weiyi Zhang, Wei Zhang, Xin Zhou, Ming Li and Jinlai Miao

Toxins 2017, 9(7), 208; https://doi.org/10.3390/toxins9070208 - 30 Jun 2017

Cited by 10 | Viewed by 4686

Abstract

Isoxanthohumol is a unique prenylflavonoid with the highest content in beer. Isoxanthohumol has multiple bioactivities and has recently received considerable attention in the scientific community. Nonetheless; its effect on drug resistant cancer cells has rarely been studied. In this paper; we investigated the [...] Read more.

Isoxanthohumol is a unique prenylflavonoid with the highest content in beer. Isoxanthohumol has multiple bioactivities and has recently received considerable attention in the scientific community. Nonetheless; its effect on drug resistant cancer cells has rarely been studied. In this paper; we investigated the synergistic effect of isoxanthohumol and doxorubicin on doxorubicin resistant MCF-7/ADR cells. Our results showed that isoxanthohumol sensitized the cytotoxic effect of doxorubicin on MCF-7/ADR cells via increased proliferation inhibition and apoptosis stimulation. Molecular mechanism studies further demonstrated that isoxanthohumol inhibited ABCB1-mediated doxorubicin efflux; stimulated the ATPase activity of ABCB1 (ATP-binding cassette sub-family B member 1); and acted as an ABCB1 substrate. Molecular docking results suggested that isoxanthohumol bound to the central transmembrane domain of ABCB1 and its binding site overlapped with the doxorubicin binding site. The present studies demonstrated that isoxanthohumol was a competitive ABCB1 inhibitor which reversed ABCB1-mediated doxorubicin resistance in MCF-7/ADR cells; and therefore could be further developed to help with overcoming ABCB1-mediated drug resistance. Full article

(This article belongs to the Section Plant Toxins)

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15 pages, 617 KiB

Open AccessArticle

Occurrence of Regulated Mycotoxins and Other Microbial Metabolites in Dried Cassava Products from Nigeria

by Adebayo B. Abass, Wasiu Awoyale, Michael Sulyok and Emmanuel O. Alamu

Toxins 2017, 9(7), 207; https://doi.org/10.3390/toxins9070207 - 29 Jun 2017

Cited by 25 | Viewed by 7001

Abstract

Dried cassava products are perceived as one of the potential sources of mycotoxin ingestion in human foods. Processing either contributes to the reduction of toxins or further exposes products to contamination by microorganisms that release metabolic toxins into the products. Thus, the prevalence [...] Read more.

Dried cassava products are perceived as one of the potential sources of mycotoxin ingestion in human foods. Processing either contributes to the reduction of toxins or further exposes products to contamination by microorganisms that release metabolic toxins into the products. Thus, the prevalence of microbial metabolites in 373 processed cassava products was investigated in Nigeria. With the use of liquid chromatography tandem-mass spectrometry (LC-MS/MS) for the constituent analysis, a few major mycotoxins (aflatoxin B₁ and G₁, fumonisin B₁ and B₂, and zearalenone) regulated in food crops by the Commission of the European Union were found at concentrations which are toxicologically acceptable in many other crops. Some bioactive compounds were detected at low concentrations in the cassava products. Therefore, the exposure of cassava consumers in Nigeria to regulated mycotoxins was estimated to be minimal. The results provide useful information regarding the probable safety of cassava products in Nigeria. Full article

(This article belongs to the Collection Understanding Mycotoxin Occurrence in Food and Feed Chains)

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13 pages, 2214 KiB

Open AccessArticle

Development and Validation of a Liquid Chromatography-Tandem Mass Spectrometry Method Coupled with Dispersive Solid-Phase Extraction for Simultaneous Quantification of Eight Paralytic Shellfish Poisoning Toxins in Shellfish

by Xianli Yang, Lei Zhou, Yanglan Tan, Xizhi Shi, Zhiyong Zhao, Dongxia Nie, Changyan Zhou and Hong Liu

Toxins 2017, 9(7), 206; https://doi.org/10.3390/toxins9070206 - 29 Jun 2017

Cited by 33 | Viewed by 6199

Abstract

In this study, a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for simultaneous determination of eight paralytic shellfish poisoning (PSP) toxins, including saxitoxin (STX), neosaxitoxin (NEO), gonyautoxins (GTX1–4) and the N-sulfo carbamoyl toxins C1 and C2, in sea shellfish. The [...] Read more.

In this study, a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for simultaneous determination of eight paralytic shellfish poisoning (PSP) toxins, including saxitoxin (STX), neosaxitoxin (NEO), gonyautoxins (GTX1–4) and the N-sulfo carbamoyl toxins C1 and C2, in sea shellfish. The samples were extracted by acetonitrile/water (80:20, v/v) with 0.1% formic and purified by dispersive solid-phase extraction (dSPE) with C18 silica and acidic alumina. Qualitative and quantitative detection for the target toxins were conducted under the multiple reaction monitoring (MRM) mode by using the positive electrospray ionization (ESI) mode after chromatographic separation on a TSK-gel Amide-80 HILIC column with water and acetonitrile. Matrix-matched calibration was used to compensate for matrix effects. The established method was further validated by determining the linearity (R² ≥ 0.9900), average recovery (81.52–116.50%), sensitivity (limits of detection (LODs): 0.33–5.52 μg·kg⁻¹; limits of quantitation (LOQs): 1.32–11.29 μg·kg⁻¹) and precision (relative standard deviation (RSD) ≤ 19.10%). The application of this proposed approach to thirty shellfish samples proved its desirable performance and sufficient capability for simultaneous determination of multiclass PSP toxins in sea foods. Full article

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

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12 pages, 6112 KiB

Open AccessArticle

Biooxidation of Ciguatoxins Leads to Species-Specific Toxin Profiles

by Tsuyoshi Ikehara, Kyoko Kuniyoshi, Naomasa Oshiro and Takeshi Yasumoto

Toxins 2017, 9(7), 205; https://doi.org/10.3390/toxins9070205 - 29 Jun 2017

Cited by 88 | Viewed by 8078

Abstract

Ciguatoxins (CTXs) contaminate fish worldwide and cause the foodborne illness ciguatera. In the Pacific, these toxins are produced by the dinoflagellate Gambierdiscus toxicus, which accumulates in fish through the food chain and undergoes oxidative modification, giving rise to numerous analogs. In this [...] Read more.

Ciguatoxins (CTXs) contaminate fish worldwide and cause the foodborne illness ciguatera. In the Pacific, these toxins are produced by the dinoflagellate Gambierdiscus toxicus, which accumulates in fish through the food chain and undergoes oxidative modification, giving rise to numerous analogs. In this study, we examined the oxidation of CTXs in vitro with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis using reference toxins, and found that CTX4A, CTX4B, and CTX3C, which are produced by the alga, are oxidized to the analogs found in fish, namely CTX1B, 52-epi-54-deoxyCTX1B, 54-deoxyCTX1B, 2-hydroxyCTX3C, and 2,3-dihydroxyCTX3C. This oxidation was catalyzed by human CYP3A4, fish liver S9 fractions, and microsomal fractions prepared from representative ciguateric fishes (Lutjanus bohar, L. monostigumus, and Oplegnathus punctatus). In addition, fish liver S9 fractions prepared from non-ciguateric fishes (L. gibbus and L. fulviflamma) in Okinawa also converted CTX4A and CTX4B to CTX1B, 54-deoxyCTX1B, and 52-epi-54-deoxyCTX1B in vitro. This is the first study to demonstrate the enzymatic oxidation of these toxins, and provides insight into the mechanism underlying the development of species-specific toxin profiles and the fate of these toxins in humans and fish. Full article

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

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10 pages, 931 KiB

Open AccessArticle

A Simple and Fast Procedure to Determine 3-Nitropropanoic Acid and 3-Nitropropanol in Freeze Dried Canadian Milkvetch (Astragalus canadensis)

by Huaizhi Liu, Suqin Shao and Mike Schellenberg

Toxins 2017, 9(7), 204; https://doi.org/10.3390/toxins9070204 - 29 Jun 2017

Cited by 4 | Viewed by 3868

Abstract

Canadian milkvetch (Astragalus canadensis) is a North American plant species in the legume family and some of this plant is fatally poisonous to livestock. The poisoning is attributed to the natural occurrence of notrotoxins, i.e., 3-nitropropanoic acid and 3-nitropropanol, present as [...] Read more.

Canadian milkvetch (Astragalus canadensis) is a North American plant species in the legume family and some of this plant is fatally poisonous to livestock. The poisoning is attributed to the natural occurrence of notrotoxins, i.e., 3-nitropropanoic acid and 3-nitropropanol, present as aglycones and conjugated forms in the plant. Those compounds cause nitrite oxidization of hemoglobin and inhibition of cellular metabolism. To determine the toxicity of the plant, it is very important to develop an analytical method for the contents of the compounds in the plant. In this study, we have successfully developed an extraction procedure followed by HPLC-UV analysis to simultaneously analyze notrotoxins. The aglycones could be released from its conjugated forms in the freeze dried plant and extracted by water at room temperature. An HPLC-UV method using a Phenomenex Kinetex 2.6 μ F5 100 Å 100 × 4.6 mm column with pH 3.5 phosphonate buffer as mobile phase have been developed and validated for the detection of the two compounds at 210 nm. This developed procedure for the analysis of 3-nitropropanoic acid and 3-nitropropanol has proven simple and efficient and it has been successfully applied for batch sample analysis. Full article

(This article belongs to the Section Plant Toxins)

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20 pages, 307 KiB

Open AccessArticle

Study on the Association among Mycotoxins and other Variables in Children with Autism

by Barbara De Santis, Maria Elisabetta Raggi, Giorgio Moretti, Francesco Facchiano, Alessandra Mezzelani, Laura Villa, Arianna Bonfanti, Alessandra Campioni, Stefania Rossi, Serena Camposeo, Sabina Soricelli, Gabriele Moracci, Francesca Debegnach, Emanuela Gregori, Francesca Ciceri, Luciano Milanesi, Anna Marabotti and Carlo Brera

Toxins 2017, 9(7), 203; https://doi.org/10.3390/toxins9070203 - 29 Jun 2017

Cited by 47 | Viewed by 9356

Abstract

Environmental factors and genetic susceptibility are implicated in the increased risk of autism spectrum disorder (ASD). Mycotoxins are agricultural contaminants of fungal origin that represent real risk factors for human health and especially for children. Thus, the main hypothesis of this work is [...] Read more.

Environmental factors and genetic susceptibility are implicated in the increased risk of autism spectrum disorder (ASD). Mycotoxins are agricultural contaminants of fungal origin that represent real risk factors for human health and especially for children. Thus, the main hypothesis of this work is that the deterioration of the clinical manifestation of autism in children may result from the exposure to mycotoxins through the consumption of contaminated food. Within a cross-sectional study, a group of autistic children (n = 172) and a group of controls (n = 61) (siblings and non-parental) were recruited in North and South Italy. All children had blood and urine samples taken, for testing some mycotoxins by a LC–MS/MS validated method. Blood samples were also tested for assessing specific IgG against food and fungal antigens and cytokines. The analyses outputs highlighted statistically significant differences comparing mycotoxins levels between (i) children groups both in urine (deoxynivalenol and de-epoxydeoxynivalenol, p = 0.0141 and p = 0.0259, respectively) and serum (aflatoxin M1, ochratoxin A and fumonisin B1, p = 0.0072, p = 0.0141 and p = 0.0061, respectively); (ii) a group of selected fungal IgGs, and IgGs against wheat and gluten and (iii) cytokines. These results suggest the need for a deeper examination of the role that mycotoxins may have on the etiology of ASD. Full article

(This article belongs to the Special Issue Exposure and Risk Assessment for Mycotoxins)

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