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Toxins, Volume 9, Issue 3 (March 2017) – 37 articles

Cover Story (view full-size image): BoNTs are diverse protein toxins. BoNT/HA is a newly identified mosaic toxin, while new toxin variants are being identified with increasing frequency. We present here the first crystal structure of the receptor-binding domain of BoNT/HA. Our structural analyses reveal that, even though BoNT/HA and the classical type A toxin (BoNT/A) have highly similar structures in this domain, BoNT/HA drastically dampens binding of a potent BoNT/A-neutralizing antibody targeting this domain via subtle genetic changes. View this paper
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1849 KiB  
Article
The Preparation and Identification of a Monoclonal Antibody against Citrinin and the Development of Detection via Indirect Competitive ELISA
by Shimuye Kalayu Yirga, Sumei Ling, Yanling Yang, Jun Yuan and Shihua Wang
Toxins 2017, 9(3), 110; https://doi.org/10.3390/toxins9030110 - 17 Mar 2017
Cited by 24 | Viewed by 5295
Abstract
Citrinin (CTN) is a hepato-nephrotoxic mycotoxin produced by fungi genera of Aspergillus, Monauscus, and Penicillium. CTN contaminates grains, fruits, juices and vegetables, and causes various toxic effects to humans and animals. It has small molecular weight, which is non-immunogenic to animals. [...] Read more.
Citrinin (CTN) is a hepato-nephrotoxic mycotoxin produced by fungi genera of Aspergillus, Monauscus, and Penicillium. CTN contaminates grains, fruits, juices and vegetables, and causes various toxic effects to humans and animals. It has small molecular weight, which is non-immunogenic to animals. Thus, CTN was conjugated to bovine serum albumin (BSA) and ovalbumin (OVA), respectively, by amide bonds using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). Mice were immunized with CTN-BSA conjugates, and spleen cells of the immunized mice were fused with Sp2/0 myeloma cells to obtain 21H27 hybriodoma cell. Ascitic fluid of hybridoma cell was produced in mice abdomen, and purified using caprylic/ammonium sulfate precipitation method. The 21H27 anti-CTN mcAb was the IgG2a antibody subclass, and cross-reactivity results indicated that anti-CTN mcAb is specific to CTN with high affinity (2.0 × 108 L/mol). Indirect competitive ELISA (ic-ELISA) results showed that the linear range of detection was 0.01–5.96 ng/mL and the IC50 was 0.28 ng/mL with a lower detection limit (LOD) of 0.01 ng/mL. The average recovery was 93.8% ± 1.6% with a coefficient variation of 1.0%–4.3%. Hence, anti-CTN mcAb secreted by 21H27 hybridoma cell was successfully produced and can be used to detect CTN contaminated feed and foodstuffs. Full article
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2542 KiB  
Article
Functional Contributions of Positive Charges in the Pore-Lining Helix 3 of the Bordetella pertussis CyaA-Hemolysin to Hemolytic Activity and Ion-Channel Opening
by Chattip Kurehong, Chalermpol Kanchanawarin, Busaba Powthongchin, Panchika Prangkio, Gerd Katzenmeier and Chanan Angsuthanasombat
Toxins 2017, 9(3), 109; https://doi.org/10.3390/toxins9030109 - 16 Mar 2017
Cited by 6 | Viewed by 5003
Abstract
The Bordetella pertussis CyaA-hemolysin (CyaA-Hly) domain was previously demonstrated to be an important determinant for hemolysis against target erythrocytes and ion-channel formation in planar lipid bilayers (PLBs). Here, net-charge variations in the pore-lining helix of thirteen related RTX cytolysins including CyaA-Hly were revealed [...] Read more.
The Bordetella pertussis CyaA-hemolysin (CyaA-Hly) domain was previously demonstrated to be an important determinant for hemolysis against target erythrocytes and ion-channel formation in planar lipid bilayers (PLBs). Here, net-charge variations in the pore-lining helix of thirteen related RTX cytolysins including CyaA-Hly were revealed by amino acid sequence alignments, reflecting their different degrees of hemolytic activity. To analyze possible functional effects of net-charge alterations on hemolytic activity and channel formation of CyaA-Hly, specific mutations were made at Gln574 or Glu581 in its pore-lining α3 of which both residues are highly conserved Lys in the three highly active RTX cytolysins (i.e., Escherichia coli α-hemolysin, Actinobacillus pleuropneumoniae toxin, and Aggregatibacter actinomycetemcomitans leukotoxin). All six constructed CyaA-Hly mutants that were over-expressed in E. coli as 126 kDa His-tagged soluble proteins were successfully purified via immobilized Ni2+-affinity chromatography. Both positive-charge substitutions (Q574K, Q574R, E581K, E581R) and negative-charge elimination (E581Q) appeared to increase the kinetics of toxin-induced hemolysis while the substitution with a negatively-charged side-chain (Q574E) completely abolished its hemolytic activity. When incorporated into PLBs under symmetrical conditions (1.0 M KCl, pH 7.4), all five mutant toxins with the increased hemolytic activity produced clearly-resolved single channels with higher open probability and longer lifetime than the wild-type toxin, albeit with a half decrease in their maximum conductance. Molecular dynamics simulations for 50 ns of a trimeric CyaA-Hly pore model comprising three α2-loop-α3 transmembrane hairpins revealed a significant role of the positive charge at both target positions in the structural stability and enlarged diameter of the simulated pore. Altogether, our present data have disclosed functional contributions of positively-charged side-chains substituted at positions Gln574 and Glu581 in the pore-lining α3 to the enhanced hemolytic activity and ion-channel opening of CyaA-Hly that actually mimics the highly-active RTX (repeat-in-toxin) cytolysins. Full article
(This article belongs to the Special Issue Adenylate Cyclase (CyaA) Toxin)
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261 KiB  
Article
Consumption of Bt Maize Pollen Containing Cry1Ie Does Not Negatively Affect Propylea japonica (Thunberg) (Coleoptera: Coccinellidae)
by Yonghui Li, Yanmin Liu, Xinming Yin, Jörg Romeis, Xinyuan Song, Xiuping Chen, Lili Geng, Yufa Peng and Yunhe Li
Toxins 2017, 9(3), 108; https://doi.org/10.3390/toxins9030108 - 16 Mar 2017
Cited by 17 | Viewed by 4891
Abstract
Propylea japonica (Thunberg) (Coleoptera: Coccinellidae) are prevalent predators and pollen feeders in East Asian maize fields. They are therefore indirectly (via prey) and directly (via pollen) exposed to Cry proteins within Bt-transgenic maize fields. The effects of Cry1Ie-producing transgenic maize pollen on [...] Read more.
Propylea japonica (Thunberg) (Coleoptera: Coccinellidae) are prevalent predators and pollen feeders in East Asian maize fields. They are therefore indirectly (via prey) and directly (via pollen) exposed to Cry proteins within Bt-transgenic maize fields. The effects of Cry1Ie-producing transgenic maize pollen on the fitness of P. japonica was assessed using two dietary-exposure experiments in the laboratory. In the first experiment, survival, larval developmental time, adult fresh weight, and fecundity did not differ between ladybirds consuming Bt or non-Bt maize pollen. In the second experiment, none of the tested lethal and sublethal parameters of P. japonica were negatively affected when fed a rapeseed pollen-based diet containing Cry1Ie protein at 200 μg/g dry weight of diet. In contrast, the larval developmental time, adult fresh weight, and fecundity of P. japonica were significantly adversely affected when fed diet containing the positive control compound E-64. In both experiments, the bioactivity of the Cry1Ie protein in the food sources was confirmed by bioassays with a Cry1Ie-sensitive lepidopteran species. These results indicated that P. japonica are not affected by the consumption of Cry1Ie-expressing maize pollen and are not sensitive to the Cry1Ie protein, suggesting that the growing of Bt maize expressing Cry1Ie protein will pose a negligible risk to P. japonica. Full article
(This article belongs to the Special Issue The Insecticidal Bacterial Toxins in Modern Agriculture)
2109 KiB  
Review
The Molecular Basis of Toxins’ Interactions with Intracellular Signaling via Discrete Portals
by Adi Lahiani, Ephraim Yavin and Philip Lazarovici
Toxins 2017, 9(3), 107; https://doi.org/10.3390/toxins9030107 - 16 Mar 2017
Cited by 22 | Viewed by 9616
Abstract
An understanding of the molecular mechanisms by which microbial, plant or animal-secreted toxins exert their action provides the most important element for assessment of human health risks and opens new insights into therapies addressing a plethora of pathologies, ranging from neurological disorders to [...] Read more.
An understanding of the molecular mechanisms by which microbial, plant or animal-secreted toxins exert their action provides the most important element for assessment of human health risks and opens new insights into therapies addressing a plethora of pathologies, ranging from neurological disorders to cancer, using toxinomimetic agents. Recently, molecular and cellular biology dissecting tools have provided a wealth of information on the action of these diverse toxins, yet, an integrated framework to explain their selective toxicity is still lacking. In this review, specific examples of different toxins are emphasized to illustrate the fundamental mechanisms of toxicity at different biochemical, molecular and cellular- levels with particular consideration for the nervous system. The target of primary action has been highlighted and operationally classified into 13 sub-categories. Selected examples of toxins were assigned to each target category, denominated as portal, and the modulation of the different portal’s signaling was featured. The first portal encompasses the plasma membrane lipid domains, which give rise to pores when challenged for example with pardaxin, a fish toxin, or is subject to degradation when enzymes of lipid metabolism such as phospholipases A2 (PLA2) or phospholipase C (PLC) act upon it. Several major portals consist of ion channels, pumps, transporters and ligand gated ionotropic receptors which many toxins act on, disturbing the intracellular ion homeostasis. Another group of portals consists of G-protein-coupled and tyrosine kinase receptors that, upon interaction with discrete toxins, alter second messengers towards pathological levels. Lastly, subcellular organelles such as mitochondria, nucleus, protein- and RNA-synthesis machineries, cytoskeletal networks and exocytic vesicles are also portals targeted and deregulated by other diverse group of toxins. A fundamental concept can be drawn from these seemingly different toxins with respect to the site of action and the secondary messengers and signaling cascades they trigger in the host. While the interaction with the initial portal is largely determined by the chemical nature of the toxin, once inside the cell, several ubiquitous second messengers and protein kinases/ phosphatases pathways are impaired, to attain toxicity. Therefore, toxins represent one of the most promising natural molecules for developing novel therapeutics that selectively target the major cellular portals involved in human physiology and diseases. Full article
(This article belongs to the Section Animal Venoms)
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Article
Simultaneous Determination of Multi-Mycotoxins in Cereal Grains Collected from South Korea by LC/MS/MS
by Dong-Ho Kim, Sung-Yong Hong, Jea Woo Kang, Sung Min Cho, Kyu Ri Lee, Tae Kyung An, Chan Lee and Soo Hyun Chung
Toxins 2017, 9(3), 106; https://doi.org/10.3390/toxins9030106 - 16 Mar 2017
Cited by 67 | Viewed by 8439
Abstract
An improved analytical method compared with conventional ones was developed for simultaneous determination of 13 mycotoxins (deoxynivalenol, nivalenol, 3-acetylnivalenol, aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, fumonisin B1, fumonisin B2, [...] Read more.
An improved analytical method compared with conventional ones was developed for simultaneous determination of 13 mycotoxins (deoxynivalenol, nivalenol, 3-acetylnivalenol, aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, fumonisin B1, fumonisin B2, T-2, HT-2, zearalenone, and ochratoxin A) in cereal grains by liquid chromatography-tandem mass spectrometry (LC/MS/MS) after a single immunoaffinity column clean-up. The method showed a good linearity, sensitivity, specificity, and accuracy in mycotoxin determination by LC/MS/MS. The levels of 13 mycotoxins in 5 types of commercial grains (brown rice, maize, millet, sorghum, and mixed cereal) from South Korea were determined in a total of 507 cereal grains. Mycotoxins produced from Fusarium sp. (fumonisins, deoxynivalenol, nivalenol, and zearalenone) were more frequently (more than 5%) and concurrently detected in all cereal grains along with higher mean levels (4.3–161.0 ng/g) in positive samples than other toxins such as aflatoxins and ochratoxin A (less than 9% and below 5.2 ng/g in positive samples) from other fungal species. Full article
(This article belongs to the Special Issue LC-MS/MS Method for Mycotoxin Analysis)
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2217 KiB  
Article
Cubozoan Sting-Site Seawater Rinse, Scraping, and Ice Can Increase Venom Load: Upending Current First Aid Recommendations
by Angel Anne Yanagihara and Christie L. Wilcox
Toxins 2017, 9(3), 105; https://doi.org/10.3390/toxins9030105 - 15 Mar 2017
Cited by 20 | Viewed by 39879
Abstract
Cnidarian envenomations are the leading cause of severe and lethal human sting injuries from marine life. The total amount of venom discharged into sting-site tissues, sometimes referred to as “venom load”, has been previously shown to correlate with tentacle contact length and sequelae [...] Read more.
Cnidarian envenomations are the leading cause of severe and lethal human sting injuries from marine life. The total amount of venom discharged into sting-site tissues, sometimes referred to as “venom load”, has been previously shown to correlate with tentacle contact length and sequelae severity. Since <1% of cnidae discharge upon initial tentacle contact, effective and safe removal of adherent tentacles is of paramount importance in the management of life-threatening cubozoan stings. We evaluated whether common rinse solutions or scraping increased venom load as measured in a direct functional assay of venom activity (hemolysis). Scraping significantly increased hemolysis by increasing cnidae discharge. For Alatina alata, increases did not occur if the tentacles were first doused with vinegar or if heat was applied. However, in Chironex fleckeri, vinegar dousing and heat treatment were less effective, and the best outcomes occurred with the use of venom-inhibiting technologies (Sting No More® products). Seawater rinsing, considered a “no-harm” alternative, significantly increased venom load. The application of ice severely exacerbated A. alata stings, but had a less pronounced effect on C. fleckeri stings, while heat application markedly reduced hemolysis for both species. Our results do not support scraping or seawater rinsing to remove adherent tentacles. Full article
(This article belongs to the Section Animal Venoms)
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3171 KiB  
Article
Ameliorative Effects of Neutral Electrolyzed Water on Growth Performance, Biochemical Constituents, and Histopathological Changes in Turkey Poults during Aflatoxicosis
by Denise Gómez-Espinosa, Francisco Javier Cervantes-Aguilar, Juan Carlos Del Río-García, Tania Villarreal-Barajas, Alma Vázquez-Durán and Abraham Méndez-Albores
Toxins 2017, 9(3), 104; https://doi.org/10.3390/toxins9030104 - 14 Mar 2017
Cited by 18 | Viewed by 5819
Abstract
Different in vitro and in silico approaches from our research group have demonstrated that neutral electrolyzed water (NEW) can be used to detoxify aflatoxins. The objective of this investigation was to evaluate the ability of NEW to detoxify B-aflatoxins (AFB1 and AFB [...] Read more.
Different in vitro and in silico approaches from our research group have demonstrated that neutral electrolyzed water (NEW) can be used to detoxify aflatoxins. The objective of this investigation was to evaluate the ability of NEW to detoxify B-aflatoxins (AFB1 and AFB2) in contaminated maize and to confirm detoxification in an in vivo experimental model. Batches of aflatoxin-contaminated maize were detoxified with NEW and mixed in commercial feed. A total of 240 6-day-old female large white Nicholas-700 turkey poults were randomly divided into four treatments of six replicates each (10 turkeys per replicate), which were fed ad libitum for two weeks with the following dietary treatments: (1) control feed containing aflatoxin-free maize (CONTROL); (2) feed containing the aflatoxin-contaminated maize (AF); (3) feed containing the aflatoxin-contaminated maize detoxified with NEW (AF + NEW); and (4) control feed containing aflatoxin-free maize treated with NEW (NEW). Compared to the control groups, turkey poults of the AF group significantly reduced body weight gain and increased feed conversion ratio and mortality rate; whereas turkey poults of the AF + NEW group did not present significant differences on productive parameters. In addition, alterations in serum biochemical constituents, enzyme activities, relative organ weight, gross morphological changes and histopathological studies were significantly mitigated by the aflatoxin-detoxification procedure. From these results, it is concluded that the treatment of aflatoxin-contaminated maize with NEW provided reasonable protection against the effects caused by aflatoxins in young turkey poults. Full article
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13619 KiB  
Article
How the Cobra Got Its Flesh-Eating Venom: Cytotoxicity as a Defensive Innovation and Its Co-Evolution with Hooding, Aposematic Marking, and Spitting
by Nadya Panagides, Timothy N.W. Jackson, Maria P. Ikonomopoulou, Kevin Arbuckle, Rudolf Pretzler, Daryl C. Yang, Syed A. Ali, Ivan Koludarov, James Dobson, Brittany Sanker, Angelique Asselin, Renan C. Santana, Iwan Hendrikx, Harold Van der Ploeg, Jeremie Tai-A-Pin, Romilly Van den Bergh, Harald M.I. Kerkkamp, Freek J. Vonk, Arno Naude, Morné A. Strydom, Louis Jacobsz, Nathan Dunstan, Marc Jaeger, Wayne C. Hodgson, John Miles and Bryan G. Fryadd Show full author list remove Hide full author list
Toxins 2017, 9(3), 103; https://doi.org/10.3390/toxins9030103 - 13 Mar 2017
Cited by 69 | Viewed by 33312
Abstract
The cytotoxicity of the venom of 25 species of Old World elapid snake was tested and compared with the morphological and behavioural adaptations of hooding and spitting. We determined that, contrary to previous assumptions, the venoms of spitting species are not consistently more [...] Read more.
The cytotoxicity of the venom of 25 species of Old World elapid snake was tested and compared with the morphological and behavioural adaptations of hooding and spitting. We determined that, contrary to previous assumptions, the venoms of spitting species are not consistently more cytotoxic than those of closely related non-spitting species. While this correlation between spitting and non-spitting was found among African cobras, it was not present among Asian cobras. On the other hand, a consistent positive correlation was observed between cytotoxicity and utilisation of the defensive hooding display that cobras are famous for. Hooding and spitting are widely regarded as defensive adaptations, but it has hitherto been uncertain whether cytotoxicity serves a defensive purpose or is somehow useful in prey subjugation. The results of this study suggest that cytotoxicity evolved primarily as a defensive innovation and that it has co-evolved twice alongside hooding behavior: once in the Hemachatus + Naja and again independently in the king cobras (Ophiophagus). There was a significant increase of cytotoxicity in the Asian Naja linked to the evolution of bold aposematic hood markings, reinforcing the link between hooding and the evolution of defensive cytotoxic venoms. In parallel, lineages with increased cytotoxicity but lacking bold hood patterns evolved aposematic markers in the form of high contrast body banding. The results also indicate that, secondary to the evolution of venom rich in cytotoxins, spitting has evolved three times independently: once within the African Naja, once within the Asian Naja, and once in the Hemachatus genus. The evolution of cytotoxic venom thus appears to facilitate the evolution of defensive spitting behaviour. In contrast, a secondary loss of cytotoxicity and reduction of the hood occurred in the water cobra Naja annulata, which possesses streamlined neurotoxic venom similar to that of other aquatic elapid snakes (e.g., hydrophiine sea snakes). The results of this study make an important contribution to our growing understanding of the selection pressures shaping the evolution of snake venom and its constituent toxins. The data also aid in elucidating the relationship between these selection pressures and the medical impact of human snakebite in the developing world, as cytotoxic cobras cause considerable morbidity including loss-of-function injuries that result in economic and social burdens in the tropics of Asia and sub-Saharan Africa. Full article
(This article belongs to the Special Issue Animal Venoms and Pain)
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7607 KiB  
Article
Characteristics and Lethality of a Novel Recombinant  Dermonecrotic Venom Phospholipase D from  Hemiscorpius lepturus
by Elham Torabi, Mahdi Behdani, Mohammad Hosseininejad Chafi, Reza Moazzami, Jean‐Marc Sabatier, Vahid Khalaj, Delavar Shahbazzadeh and Kamran Pooshang Bagheri
Toxins 2017, 9(3), 102; https://doi.org/10.3390/toxins9030102 - 13 Mar 2017
Cited by 16 | Viewed by 5955
Abstract
Hemoscorpius lepturus is the most medically important scorpion in Iran. The clinical signs of H. lepturus envenomation are remarkably similar to those reported for brown spiders, including dermonecrosis, hematuria, renal failure and even death. The lethality and toxicity of brown spiders’ venom have [...] Read more.
Hemoscorpius lepturus is the most medically important scorpion in Iran. The clinical signs of H. lepturus envenomation are remarkably similar to those reported for brown spiders, including dermonecrosis, hematuria, renal failure and even death. The lethality and toxicity of brown spiders’ venom have been attributed to its phospholipase D activity. This study aims to identify a phospholipase D with possible lethality and dermonecrotic activity in H. lepturus venom. In this study, a cDNA library of the venom glands was generated by Illumina RNA sequencing. Phospholipase D (PLD) from H. lepturus was characterized according to its significant similarity with PLDs from brown spiders. The main chain designated as Hl‐RecPLD1 (the first recombinant isoform of H. lepturus PLD) was cloned, expressed and purified. Sphingomyelinase, dermonecrotic and lethal activities were examined. Hl‐PLD1 showed remarkable sequence similarity and structural homology with PLDs of brown spiders. The conformation of Hl‐PLD1 was predicted as a “TIM beta/alpha‐barrel”. The lethal dose 50 (LD50) and dermonecrotic activities of Hl‐RecPLD1 were determined as 3.1 μg/mouse and 0.7 cm2 at 1 μg respectively. It is the first report indicating that a similar molecular evolutionary mechanism has occurred in both American brown spiders and this Iranian scorpion. In conclusion, Hl‐RecPLD1 is a highly active phospholipase D, which would be considered as the lethal dermonecrotic toxin in H. lepturus venom. Full article
(This article belongs to the Section Animal Venoms)
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789 KiB  
Review
Helicobacter pylori Outer Membrane Protein-Related Pathogenesis
by Yuichi Matsuo, Yasutoshi Kido and Yoshio Yamaoka
Toxins 2017, 9(3), 101; https://doi.org/10.3390/toxins9030101 - 11 Mar 2017
Cited by 59 | Viewed by 12598
Abstract
Helicobacter pylori colonizes the human stomach and induces inflammation, and in some cases persistent infection can result in gastric cancer. Attachment to the gastric mucosa is the first step in establishing bacterial colonization, and outer membrane proteins (OMPs) play a pivotal role in [...] Read more.
Helicobacter pylori colonizes the human stomach and induces inflammation, and in some cases persistent infection can result in gastric cancer. Attachment to the gastric mucosa is the first step in establishing bacterial colonization, and outer membrane proteins (OMPs) play a pivotal role in binding to human cells. Some OMP interaction molecules are known in H. pylori, and their associated host cell responses have been gradually clarified. Many studies have demonstrated that OMPs are essential to CagA translocation into gastric cells via the Type IV secretion system of H. pylori. This review summarizes the mechanisms through which H. pylori utilizes OMPs to colonize the human stomach and how OMPs cooperate with the Type IV secretion system. Full article
(This article belongs to the Special Issue H. pylori Virulence Factors in the Induction of Gastric Cancer)
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3997 KiB  
Article
Nitrate Increased Cucumber Tolerance to Fusarium Wilt by Regulating Fungal Toxin Production and Distribution
by Jinyan Zhou, Min Wang, Yuming Sun, Zechen Gu, Ruirui Wang, Asanjan Saydin, Qirong Shen and Shiwei Guo
Toxins 2017, 9(3), 100; https://doi.org/10.3390/toxins9030100 - 11 Mar 2017
Cited by 37 | Viewed by 8189
Abstract
Cucumber Fusarium wilt, induced by Fusarium oxysporum f. sp. cucumerinum (FOC), causes severe losses in cucumber yield and quality. Nitrogen (N), as the most important mineral nutrient for plants, plays a critical role in plant–pathogen interactions. Hydroponic assays were conducted to investigate the [...] Read more.
Cucumber Fusarium wilt, induced by Fusarium oxysporum f. sp. cucumerinum (FOC), causes severe losses in cucumber yield and quality. Nitrogen (N), as the most important mineral nutrient for plants, plays a critical role in plant–pathogen interactions. Hydroponic assays were conducted to investigate the effects of different N forms (NH4+ vs. NO3) and supply levels (low, 1 mM; high, 5 mM) on cucumber Fusarium wilt. The NO3-fed cucumber plants were more tolerant to Fusarium wilt compared with NH4+-fed plants, and accompanied by lower leaf temperature after FOC infection. The disease index decreased as the NO3 supply increased but increased with the NH4+ level supplied. Although the FOC grew better under high NO3 in vitro, FOC colonization and fusaric acid (FA) production decreased in cucumber plants under high NO3 supply, associated with lower leaf membrane injury. There was a positive correlation between the FA content and the FOC number or relative membrane injury. After the exogenous application of FA, less FA accumulated in the leaves under NO3 feeding, accompanied with a lower leaf membrane injury. In conclusion, higher NO3 supply protected cucumber plants against Fusarium wilt by suppressing FOC colonization and FA production in plants, and increasing the plant tolerance to FA. Full article
(This article belongs to the Special Issue Exposure and Risk Assessment for Mycotoxins)
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Article
A Biologically-Based Computational Approach to Drug Repurposing for Anthrax Infection
by Jane P. F. Bai, Theodore Sakellaropoulos and Leonidas G. Alexopoulos
Toxins 2017, 9(3), 99; https://doi.org/10.3390/toxins9030099 - 10 Mar 2017
Cited by 3 | Viewed by 6052
Abstract
Developing drugs to treat the toxic effects of lethal toxin (LT) and edema toxin (ET) produced by B. anthracis is of global interest. We utilized a computational approach to score 474 drugs/compounds for their ability to reverse the toxic effects of anthrax [...] Read more.
Developing drugs to treat the toxic effects of lethal toxin (LT) and edema toxin (ET) produced by B. anthracis is of global interest. We utilized a computational approach to score 474 drugs/compounds for their ability to reverse the toxic effects of anthrax toxins. For each toxin or drug/compound, we constructed an activity network by using its differentially expressed genes, molecular targets, and protein interactions. Gene expression profiles of drugs were obtained from the Connectivity Map and those of anthrax toxins in human alveolar macrophages were obtained from the Gene Expression Omnibus. Drug rankings were based on the ability of a drug/compound’s mode of action in the form of a signaling network to reverse the effects of anthrax toxins; literature reports were used to verify the top 10 and bottom 10 drugs/compounds identified. Simvastatin and bepridil with reported in vitro potency for protecting cells from LT and ET toxicities were computationally ranked fourth and eighth. The other top 10 drugs were fenofibrate, dihydroergotamine, cotinine, amantadine, mephenytoin, sotalol, ifosfamide, and mefloquine; literature mining revealed their potential protective effects from LT and ET toxicities. These drugs are worthy of investigation for their therapeutic benefits and might be used in combination with antibiotics for treating B. anthracis infection. Full article
(This article belongs to the Collection Anthrax Toxins)
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Article
The Influence of Low Doses of Zearalenone and T-2 Toxin on Calcitonin Gene Related Peptide-Like Immunoreactive (CGRP-LI) Neurons in the ENS of the Porcine Descending Colon
by Krystyna Makowska, Kazimierz Obremski, Lukasz Zielonka and Slawomir Gonkowski
Toxins 2017, 9(3), 98; https://doi.org/10.3390/toxins9030098 - 10 Mar 2017
Cited by 37 | Viewed by 5696
Abstract
The enteric nervous system (ENS) can undergo adaptive and reparative changes in response to physiological and pathological stimuli. These manifest primarily as alterations in the levels of active substances expressed by the enteric neuron. While it is known that mycotoxins can affect the [...] Read more.
The enteric nervous system (ENS) can undergo adaptive and reparative changes in response to physiological and pathological stimuli. These manifest primarily as alterations in the levels of active substances expressed by the enteric neuron. While it is known that mycotoxins can affect the function of the central and peripheral nervous systems, knowledge about their influence on the ENS is limited. Therefore, the aim of the present study was to investigate the influence of low doses of zearalenone (ZEN) and T-2 toxin on calcitonin gene related peptide-like immunoreactive (CGRP-LI) neurons in the ENS of the porcine descending colon using a double immunofluorescence technique. Both mycotoxins led to an increase in the percentage of CGRP-LI neurons in all types of enteric plexuses and changed the degree of co-localization of CGRP with other neuronal active substances, such as substance P, galanin, nitric oxide synthase, and cocaine- and amphetamine-regulated transcript peptide. The obtained results demonstrate that even low doses of ZEN and T-2 can affect living organisms and cause changes in the neurochemical profile of enteric neurons. Full article
(This article belongs to the Section Mycotoxins)
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1960 KiB  
Article
Plasma-Based Degradation of Mycotoxins Produced by Fusarium, Aspergillus and Alternaria Species
by Lars Ten Bosch, Katharina Pfohl, Georg Avramidis, Stephan Wieneke, Wolfgang Viöl and Petr Karlovsky
Toxins 2017, 9(3), 97; https://doi.org/10.3390/toxins9030097 - 10 Mar 2017
Cited by 118 | Viewed by 11100
Abstract
The efficacy of cold atmospheric pressure plasma (CAPP) with ambient air as working gas for the degradation of selected mycotoxins was studied. Deoxynivalenol, zearalenone, enniatins, fumonisin B1, and T2 toxin produced by Fusarium spp., sterigmatocystin produced by Aspergillus spp. and AAL toxin produced [...] Read more.
The efficacy of cold atmospheric pressure plasma (CAPP) with ambient air as working gas for the degradation of selected mycotoxins was studied. Deoxynivalenol, zearalenone, enniatins, fumonisin B1, and T2 toxin produced by Fusarium spp., sterigmatocystin produced by Aspergillus spp. and AAL toxin produced by Alternaria alternata were used. The kinetics of the decay of mycotoxins exposed to plasma discharge was monitored. All pure mycotoxins exposed to CAPP were degraded almost completely within 60 s. Degradation rates varied with mycotoxin structure: fumonisin B1 and structurally related AAL toxin were degraded most rapidly while sterigmatocystin exhibited the highest resistance to degradation. As compared to pure compounds, the degradation rates of mycotoxins embedded in extracts of fungal cultures on rice were reduced to a varying extent. Our results show that CAPP efficiently degrades pure mycotoxins, the degradation rates vary with mycotoxin structure, and the presence of matrix slows down yet does not prevent the degradation. CAPP appears promising for the decontamination of food commodities with mycotoxins confined to or enriched on surfaces such as cereal grains. Full article
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936 KiB  
Article
Individual and Combined Cytotoxic Effects of   Co‐Occurring Deoxynivalenol Family Mycotoxins on  Human Gastric Epithelial Cells
by Yunxia Yang, Song Yu, Yanglan Tan, Na Liu and Aibo Wu 
Toxins 2017, 9(3), 96; https://doi.org/10.3390/toxins9030096 - 09 Mar 2017
Cited by 49 | Viewed by 5060
Abstract
Mycotoxin contamination is a significant health concern for human beings, but health risk assessments are usually based on one single mycotoxin, which might neglect the additive or competitive interactions between co‐occurring mycotoxins [...] Full article
(This article belongs to the Special Issue Exposure and Risk Assessment for Mycotoxins)
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1347 KiB  
Article
Multiple Stressors at the Land-Sea Interface: Cyanotoxins at the Land-Sea Interface in the Southern California Bight
by Avery O. Tatters, Meredith D.A. Howard, Carey Nagoda, Lilian Busse, Alyssa G. Gellene and David A. Caron
Toxins 2017, 9(3), 95; https://doi.org/10.3390/toxins9030095 - 09 Mar 2017
Cited by 29 | Viewed by 7974
Abstract
Blooms of toxic cyanobacteria in freshwater ecosystems have received considerable attention in recent years, but their occurrence and potential importance at the land-sea interface has not been widely recognized. Here we present the results of a survey of discrete samples conducted in more [...] Read more.
Blooms of toxic cyanobacteria in freshwater ecosystems have received considerable attention in recent years, but their occurrence and potential importance at the land-sea interface has not been widely recognized. Here we present the results of a survey of discrete samples conducted in more than fifty brackish water sites along the coastline of southern California. Our objectives were to characterize cyanobacterial community composition and determine if specific groups of cyanotoxins (anatoxins, cylindrospermopsins, microcystins, nodularins, and saxitoxins) were present. We report the identification of numerous potentially harmful taxa and the co-occurrence of multiple toxins, previously undocumented, at several locations. Our findings reveal a potential health concern based on the range of organisms present and the widespread prevalence of recognized toxic compounds. Our results raise concerns for recreation, harvesting of finfish and shellfish, and wildlife and desalination operations, highlighting the need for assessments and implementation of monitoring programs. Such programs appear to be particularly necessary in regions susceptible to urban influence. Full article
(This article belongs to the Collection Marine and Freshwater Toxins)
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540 KiB  
Article
Implementing the Bruker MALDI Biotyper in the Public Health Laboratory for C. botulinum Neurotoxin Detection
by Michael J. Perry, Dominick A. Centurioni, Stephen W. Davis, George E. Hannett, Kimberlee A. Musser and Christina T. Egan
Toxins 2017, 9(3), 94; https://doi.org/10.3390/toxins9030094 - 09 Mar 2017
Cited by 13 | Viewed by 6498
Abstract
Currently, the gold standard method for active botulinum neurotoxin (BoNT) detection is the mouse bioassay (MBA). A Centers for Disease Control and Prevention-developed mass spectrometry (MS)-based assay that detects active BoNT was successfully validated and implemented in a public health laboratory in clinical [...] Read more.
Currently, the gold standard method for active botulinum neurotoxin (BoNT) detection is the mouse bioassay (MBA). A Centers for Disease Control and Prevention-developed mass spectrometry (MS)-based assay that detects active BoNT was successfully validated and implemented in a public health laboratory in clinical matrices using the Bruker MALDI-TOF MS (Matrix-assisted laser desorption ionization–time of flight mass spectrometry) Biotyper. For the first time, a direct comparison with the MBA was performed to determine MS-based assay sensitivity using the Bruker MALDI Biotyper. Mice were injected with BoNT/A, /B, /E, and /F at concentrations surrounding the established MS assay limit of detection (LOD) and analyzed simultaneously. For BoNT/B, /E, and /F, MS assay sensitivity was equivalent or better than the MBA at 25, 0.3, and 8.8 mLD50, respectively. BoNT/A was detected by the MBA between 1.8 and 18 mLD50, somewhat more sensitive than the MS method of 18 mLD50. Studies were performed to compare assay performance in clinical specimens. For all tested specimens, the MS method rapidly detected BoNT activity and serotype in agreement with, or in the absence of, results from the MBA. We demonstrate that the MS assay can generate reliable, rapid results while eliminating the need for animal testing. Full article
(This article belongs to the Collection Rapid Detection of Bacterial Toxins)
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798 KiB  
Review
In Vivo Crystallization of Three-Domain Cry Toxins
by Rooma Adalat, Faiza Saleem, Neil Crickmore, Shagufta Naz and Abdul Rauf Shakoori
Toxins 2017, 9(3), 80; https://doi.org/10.3390/toxins9030080 - 09 Mar 2017
Cited by 21 | Viewed by 6863
Abstract
Bacillus thuringiensis (Bt) is the most successful, environmentally-friendly, and intensively studied microbial insecticide. The major characteristic of Bt is the production of proteinaceous crystals containing toxins with specific activity against many pests including dipteran, lepidopteran, and coleopteran insects, as well as nematodes, protozoa, [...] Read more.
Bacillus thuringiensis (Bt) is the most successful, environmentally-friendly, and intensively studied microbial insecticide. The major characteristic of Bt is the production of proteinaceous crystals containing toxins with specific activity against many pests including dipteran, lepidopteran, and coleopteran insects, as well as nematodes, protozoa, flukes, and mites. These crystals allow large quantities of the protein toxins to remain stable in the environment until ingested by a susceptible host. It has been previously established that 135 kDa Cry proteins have a crystallization domain at their C-terminal end. In the absence of this domain, Cry proteins often need helper proteins or other factors for crystallization. In this review, we classify the Cry proteins based on their requirements for crystallization. Full article
(This article belongs to the Special Issue The Insecticidal Bacterial Toxins in Modern Agriculture)
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2673 KiB  
Article
Crystal Structure of the Receptor-Binding Domain of Botulinum Neurotoxin Type HA, Also Known as Type FA or H
by Guorui Yao, Kwok-ho Lam, Kay Perry, Jasmin Weisemann, Andreas Rummel and Rongsheng Jin
Toxins 2017, 9(3), 93; https://doi.org/10.3390/toxins9030093 - 08 Mar 2017
Cited by 23 | Viewed by 6238
Abstract
Botulinum neurotoxins (BoNTs), which have been exploited as cosmetics and muscle-disorder treatment medicines for decades, are well known for their extreme neurotoxicity to humans. They pose a potential bioterrorism threat because they cause botulism, a flaccid muscular paralysis-associated disease that requires immediate antitoxin [...] Read more.
Botulinum neurotoxins (BoNTs), which have been exploited as cosmetics and muscle-disorder treatment medicines for decades, are well known for their extreme neurotoxicity to humans. They pose a potential bioterrorism threat because they cause botulism, a flaccid muscular paralysis-associated disease that requires immediate antitoxin treatment and intensive care over a long period of time. In addition to the existing seven established BoNT serotypes (BoNT/A–G), a new mosaic toxin type termed BoNT/HA (aka type FA or H) was reported recently. Sequence analyses indicate that the receptor-binding domain (HC) of BoNT/HA is ~84% identical to that of BoNT/A1. However, BoNT/HA responds differently to some potent BoNT/A-neutralizing antibodies (e.g., CR2) that target the HC. Therefore, it raises a serious concern as to whether BoNT/HA poses a new threat to our biosecurity. In this study, we report the first high-resolution crystal structure of BoNT/HA-HC at 1.8 Å. Sequence and structure analyses reveal that BoNT/HA and BoNT/A1 are different regarding their binding to cell-surface receptors including both polysialoganglioside (PSG) and synaptic vesicle glycoprotein 2 (SV2). Furthermore, the new structure also provides explanations for the ~540-fold decreased affinity of antibody CR2 towards BoNT/HA compared to BoNT/A1. Taken together, these new findings advance our understanding of the structure and function of this newly identified toxin at the molecular level, and pave the way for the future development of more effective countermeasures. Full article
(This article belongs to the Section Bacterial Toxins)
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2422 KiB  
Article
A Greener, Quick and Comprehensive Extraction Approach for LC-MS of Multiple Mycotoxins
by Andreas Breidbach
Toxins 2017, 9(3), 91; https://doi.org/10.3390/toxins9030091 - 07 Mar 2017
Cited by 13 | Viewed by 6963
Abstract
In food/feed control, mycotoxin analysis is often still performed “one analyte at a time”. Here a method is presented which aims at making mycotoxin analysis environmentally friendlier through replacing acetonitrile by ethyl acetate and reducing chemical waste production by analyzing four mycotoxins together, [...] Read more.
In food/feed control, mycotoxin analysis is often still performed “one analyte at a time”. Here a method is presented which aims at making mycotoxin analysis environmentally friendlier through replacing acetonitrile by ethyl acetate and reducing chemical waste production by analyzing four mycotoxins together, forgoing sample extract clean-up, and minimizing solvent consumption. For this, 2 g of test material were suspended in 8 mL water and 16 mL ethyl acetate were added. Extraction was accelerated through sonication for 30 min and subsequent addition of 8 g sodium sulfate. After centrifugation, 500 µL supernatant were spiked with isotopologues, dried down, reconstituted in mobile phase, and measured with LC-MS. The method was validated in-house and through a collaborative study and the performance was fit-for-purpose. Repeatability relative standard deviation (RSDs) between 16% at low and 4% at higher contaminations were obtained. The reproducibility RSDs were mostly between 12% and 32%. The trueness of results for T-2 toxin and Zearalenone were not different from 100%, for Deoxynivalenol and HT-2 toxin they were larger than 89%. The extraction was also adapted to a quick screening of Aflatoxin B1 in maize by flow-injection–mass spectrometry. Semi-quantitative results were obtained through standard addition and scan-based ion ratio calculations. The method proved to be a viable greener and quicker alternative to existing methods. Full article
(This article belongs to the Special Issue LC-MS/MS Method for Mycotoxin Analysis)
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530 KiB  
Review
Toxic Dimethylarginines: Asymmetric  Dimethylarginine (ADMA) and Symmetric  Dimethylarginine (SDMA)
by You‐Lin Tain and Chien‐Ning Hsu
Toxins 2017, 9(3), 92; https://doi.org/10.3390/toxins9030092 - 06 Mar 2017
Cited by 167 | Viewed by 11837
Abstract
Asymmetric and symmetric dimethylarginine (ADMA and SDMA, respectively) are toxic, non‐proteinogenic amino acids formed by post‐translational modification and are uremic toxins that inhibit nitric oxide (NO) production and play multifunctional roles in many human diseases. Both ADMA and SDMA have emerged as strong [...] Read more.
Asymmetric and symmetric dimethylarginine (ADMA and SDMA, respectively) are toxic, non‐proteinogenic amino acids formed by post‐translational modification and are uremic toxins that inhibit nitric oxide (NO) production and play multifunctional roles in many human diseases. Both ADMA and SDMA have emerged as strong predictors of cardiovascular events and death in a range of illnesses. Major progress has been made in research on ADMA‐lowering therapies in animal studies; however, further studies are required to fill the translational gap between animal models and clinical trials in order to treat human diseases related to elevated ADMA/SDMA levels. Here, we review the reported impacts of ADMA and SDMA on human health and disease, focusing on the synthesis and metabolism of ADMA and SDMA; the pathophysiological roles of these dimethylarginines; clinical conditions and animal models associated with elevated ADMA and SDMA levels; and potential therapies against ADMA and SDMA. There is currently no specific pharmacological therapy for lowering the levels and counteracting the deleterious effects of ADMA and SDMA. A better understanding of the mechanisms underlying the impact of ADMA and SDMA on a wide range of human diseases is essential to the development of specific therapies against diseases related to ADMA and SDMA. Full article
(This article belongs to the Special Issue Toxic Non-Proteinogenic Amino Acids (NPAA))
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1881 KiB  
Article
Tetracycline Reduces Kidney Damage Induced by Loxosceles Spider Venom
by Cinthya Kimori Okamoto, Carmen W. Van den Berg, Mizuno Masashi, Rute M. Gonçalves-de-Andrade and Denise V. Tambourgi
Toxins 2017, 9(3), 90; https://doi.org/10.3390/toxins9030090 - 02 Mar 2017
Cited by 14 | Viewed by 5363
Abstract
Envenomation by Loxosceles spider can result in two clinical manifestations: cutaneous and systemic loxoscelism, the latter of which includes renal failure. Although incidence of renal failure is low, it is the main cause of death, occurring mainly in children. The sphingomyelinase D (SMase [...] Read more.
Envenomation by Loxosceles spider can result in two clinical manifestations: cutaneous and systemic loxoscelism, the latter of which includes renal failure. Although incidence of renal failure is low, it is the main cause of death, occurring mainly in children. The sphingomyelinase D (SMase D) is the main component in Loxosceles spider venom responsible for local and systemic manifestations. This study aimed to investigate the toxicity of L. intermedia venom and SMase D on kidney cells, using both In vitro and in vivo models, and the possible involvement of endogenous metalloproteinases (MMP). Results demonstrated that venom and SMase D are able to cause death of human kidney cells by apoptosis, concomitant with activation and secretion of extracellular matrix metalloproteases, MMP-2 and MMP-9. Furthermore, cell death and MMP synthesis and secretion can be prevented by tetracycline. In a mouse model of systemic loxoscelism, Loxosceles venom-induced kidney failure was observed, which was abrogated by administration of tetracycline. These results indicate that MMPs may play an important role in Loxosceles venom-induced kidney injury and that tetracycline administration may be useful in the treatment of human systemic loxoscelism. Full article
(This article belongs to the Section Animal Venoms)
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2553 KiB  
Communication
Microcystin‐LR Detected in a Low Molecular Weight  Fraction from a Crude Extract of Zoanthus sociatus
by Dany Domínguez‐Pérez, Armando Alexei Rodríguez, Hugo Osorio, Joana Azevedo, Olga Castañeda, Vítor Vasconcelos and Agostinho Antunes
Toxins 2017, 9(3), 89; https://doi.org/10.3390/toxins9030089 - 01 Mar 2017
Cited by 5 | Viewed by 4714
Abstract
Cnidarian constitutes a great source of bioactive compounds. However, research involving peptides from organisms belonging to the order Zoanthidea has received very little attention, contrasting to the numerous studies of the order Actiniaria, from which hundreds of toxic peptides and proteins have been [...] Read more.
Cnidarian constitutes a great source of bioactive compounds. However, research involving peptides from organisms belonging to the order Zoanthidea has received very little attention, contrasting to the numerous studies of the order Actiniaria, from which hundreds of toxic peptides and proteins have been reported. In this work, we performed a mass spectrometry analysis of a low molecular weight (LMW) fraction previously reported as lethal to mice. The low molecular weight (LMW) fraction was obtained by gel filtration of a Zoanthus sociatus (order Zoanthidea) crude extract with a Sephadex G‐50, and then analyzed by matrix‐assisted laser desorption/ionization time‐of‐flight/time‐of‐flight (MALDI‐TOF/TOF) mass spectrometry (MS) in positive ion reflector mode from m/z 700 to m/z 4000. Afterwards, some of the most intense and representative MS ions were fragmented by MS/MS with no significant results obtained by Protein Pilot protein identification software and the Mascot algorithm search. However, microcystin masses were detected by mass‐matching against libraries of non‐ribosomal peptide database (NORINE). Subsequent reversed‐phase C18 HPLC (in isocratic elution mode) and mass spectrometry analyses corroborated the presence of the cyanotoxin Microcystin‐LR (MC‐LR). To the best of our knowledge, this finding constitutes the first report of MC‐LR in Z. sociatus, and one of the few evidences of such cyanotoxin in cnidarians. Full article
(This article belongs to the Section Marine and Freshwater Toxins)
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2584 KiB  
Article
Effects of pH and Temperature on the Stability of Fumonisins in Maize Products
by Marcin Bryła, Agnieszka Waśkiewicz, Krystyna Szymczyk and Renata Jędrzejczak
Toxins 2017, 9(3), 88; https://doi.org/10.3390/toxins9030088 - 01 Mar 2017
Cited by 31 | Viewed by 6147
Abstract
This paper is a study of the stability of fumonisins in dough based on maize flour prepared in a phosphate buffer with a pH of 3.5, 5.5 or 7.5 and baked at a temperature within the range of 100–250 °C. Buffers with various [...] Read more.
This paper is a study of the stability of fumonisins in dough based on maize flour prepared in a phosphate buffer with a pH of 3.5, 5.5 or 7.5 and baked at a temperature within the range of 100–250 °C. Buffers with various pH values were tested, since it is well-known that pH may significantly influence interactions of fumonisins with other substances. A standard analytical procedure was used to determine the concentration of free fumonisins. Hydrolysis in an alkaline medium was then applied to reveal the hidden forms, while the total fumonisins concentations was determined in another measurement. The total concentration of fumonisins was statistically higher in pH = 3.5 and pH = 5.5 than the concentration of free fumonisins; no similar difference was found at pH = 7.5. The applied phosphate buffer pH 7.5 may enhance solubility of fumonisins, which would increase extraction efficiency of free analytes, thereby decreasing the difference between concentrations of total and free fumonisins. Hydrolysed B1 fumonisin (HFB1) and partially hydrolysed B1 fumonisin (isomers a and b: PHFB1a and PHFB1b, respectively) were the main investigated substances. For baking temperatures below 220 °C, fumonisins were slightly more stable for pH = 5.5 than for pH = 3.5 and pH = 7.5. In both of these latter cases, the concentration of partially hydrolysed fumonisins grew initially (up to 200 °C) with an increase in the baking temperature, and then dropped. Similar behaviour was observed for free HFB1, which may suggest the following fumonisin degradation mechanism: initially, the tricarballylic acid (TCA) groups are removed from the molecules, and next, the HFB1 molecules disintegrate. Full article
(This article belongs to the Collection Understanding Mycotoxin Occurrence in Food and Feed Chains)
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4655 KiB  
Article
Identification of the Anti-Aflatoxinogenic Activity of Micromeria graeca and Elucidation of Its Molecular Mechanism in Aspergillus flavus
by Rhoda El Khoury, Isaura Caceres, Olivier Puel, Sylviane Bailly, Ali Atoui, Isabelle P. Oswald, André El Khoury and Jean-Denis Bailly
Toxins 2017, 9(3), 87; https://doi.org/10.3390/toxins9030087 - 01 Mar 2017
Cited by 30 | Viewed by 10264
Abstract
Of all the food-contaminating mycotoxins, aflatoxins, and most notably aflatoxin B1 (AFB1), are found to be the most toxic and economically costly. Green farming is striving to replace fungicides and develop natural preventive strategies to minimize crop contamination by these [...] Read more.
Of all the food-contaminating mycotoxins, aflatoxins, and most notably aflatoxin B1 (AFB1), are found to be the most toxic and economically costly. Green farming is striving to replace fungicides and develop natural preventive strategies to minimize crop contamination by these toxic fungal metabolites. In this study, we demonstrated that an aqueous extract of the medicinal plant Micromeria graeca—known as hyssop—completely inhibits aflatoxin production by Aspergillus flavus without reducing fungal growth. The molecular inhibitory mechanism was explored by analyzing the expression of 61 genes, including 27 aflatoxin biosynthesis cluster genes and 34 secondary metabolism regulatory genes. This analysis revealed a three-fold down-regulation of aflR and aflS encoding the two internal cluster co-activators, resulting in a drastic repression of all aflatoxin biosynthesis genes. Hyssop also targeted fifteen regulatory genes, including veA and mtfA, two major global-regulating transcription factors. The effect of this extract is also linked to a transcriptomic variation of several genes required for the response to oxidative stress such as msnA, srrA, catA, cat2, sod1, mnsod, and stuA. In conclusion, hyssop inhibits AFB1 synthesis at the transcriptomic level. This aqueous extract is a promising natural-based solution to control AFB1 contamination. Full article
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3031 KiB  
Article
Interaction of Type IV Toxin/Antitoxin Systems in Cryptic Prophages of Escherichia coli K-12
by Zhongling Wen, Pengxia Wang, Chenglong Sun, Yunxue Guo and Xiaoxue Wang
Toxins 2017, 9(3), 77; https://doi.org/10.3390/toxins9030077 - 01 Mar 2017
Cited by 26 | Viewed by 6766
Abstract
Toxin/antitoxin (TA) systems are widespread in prokaryotic chromosomes and in mobile genetic elements including plasmids and prophages. The first characterized Type IV TA system CbtA/CbeA was found in cryptic prophage CP4-44 in Escherichia coli K-12. Two homologous TA loci of CbtA/CbeA also reside [...] Read more.
Toxin/antitoxin (TA) systems are widespread in prokaryotic chromosomes and in mobile genetic elements including plasmids and prophages. The first characterized Type IV TA system CbtA/CbeA was found in cryptic prophage CP4-44 in Escherichia coli K-12. Two homologous TA loci of CbtA/CbeA also reside in cryptic prophages of E. coli K-12, YkfI/YafW in CP4-6 and YpjF/YfjZ in CP4-57. In this study, we demonstrated that YkfI and YpjF inhibited cell growth and led to the formation of “lemon-shaped” cells. Prolonged overproduction of YkfI led to the formation of “gourd-shaped” cells and immediate cell lysis. YafW and YfjZ can neutralize the toxicity of YkfI or YpjF. Furthermore, we found that YkfI and YpjF interacted with cell division protein FtsZ in E. coli, but ectopic expression in Pseudomonas and Shewanella did not cause the formation of “lemon-shaped” cells. Moreover, deletion of all of the three toxin genes together decreased resistance to oxidative stress and deletion of the antitoxin genes increased early biofilm formation. Collectively, these results demonstrated that the homologous Type IV TA systems in E. coli may target cell division protein FtsZ in E. coli and may have different physiological functions in E. coli. Full article
(This article belongs to the Section Bacterial Toxins)
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1041 KiB  
Article
Sterigmatocystin Occurrence in Paddy and Processed Rice Produced in Italy in the Years 2014–2015 and Distribution in Milled Rice Fractions
by Terenzio Bertuzzi, Marco Romani, Silvia Rastelli, Annalisa Mulazzi and Amedeo Pietri
Toxins 2017, 9(3), 86; https://doi.org/10.3390/toxins9030086 - 28 Feb 2017
Cited by 19 | Viewed by 5459
Abstract
The occurrence of sterigmatocystin (STC) in paddy and processed rice samples produced in Italy was surveyed. After extraction and purification, STC was analysed using HPLC-MS/MS. STC was detected in all paddy rice samples (n = 49), in the range 0.29–15.85 μg·kg−1 [...] Read more.
The occurrence of sterigmatocystin (STC) in paddy and processed rice samples produced in Italy was surveyed. After extraction and purification, STC was analysed using HPLC-MS/MS. STC was detected in all paddy rice samples (n = 49), in the range 0.29–15.85 μg·kg−1. As regards processed rice, a widespread contamination was found in brown and parboiled rice. All the brown rice samples were contaminated between 0.12 and 1.32 μg·kg−1; for parboiled rice, the incidence was 90.9% and the maximum level was 1.09 μg·kg−1. The contamination in white rice was significantly lower (p < 0.01). The STC distribution in different rice fractions, obtained by the de-hulling and polishing processes, was evaluated. After de-hulling, the STC percentage remaining in brown rice was in the range 21.2%–30.8%. The polishing process, from brown to white rice, caused another remarkable decrease of contamination; the STC remaining in white rice was 2.2%–8.3% of the amount found in paddy rice. Full article
(This article belongs to the Collection Understanding Mycotoxin Occurrence in Food and Feed Chains)
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463 KiB  
Review
Do Plant-Bound Masked Mycotoxins Contribute to Toxicity?
by Silvia W. Gratz
Toxins 2017, 9(3), 85; https://doi.org/10.3390/toxins9030085 - 28 Feb 2017
Cited by 48 | Viewed by 8277
Abstract
Masked mycotoxins are plant metabolites of mycotoxins which co-contaminate common cereal crops. Since their discovery, the question has arisen if they contribute to toxicity either directly or indirectly through the release of the parent mycotoxins. Research in this field is rapidly emerging and [...] Read more.
Masked mycotoxins are plant metabolites of mycotoxins which co-contaminate common cereal crops. Since their discovery, the question has arisen if they contribute to toxicity either directly or indirectly through the release of the parent mycotoxins. Research in this field is rapidly emerging and the aim of this review is to summarize the latest knowledge on the fate of masked mycotoxins upon ingestion. Fusarium mycotoxins are the most prevalent masked mycotoxins and evidence is mounting that DON3Glc and possibly other masked trichothecenes are stable in conditions prevailing in the upper gut and are not absorbed intact. DON3Glc is also not toxic per se, but is hydrolyzed by colonic microbes and further metabolized to DOM-1 in some individuals. Masked zearalenone is rather more bio-reactive with some evidence on gastric and small intestinal hydrolysis as well as hydrolysis by intestinal epithelium and components of blood. Microbial hydrolysis of ZEN14Glc is almost instantaneous and further metabolism also occurs. Identification of zearalenone metabolites and their fate in the colon are still missing as is further clarification on whether or not masked zearalenone is hydrolyzed by mammalian cells. New masked mycotoxins continuously emerge and it is crucial that we gain detailed understanding of their individual metabolic fate in the body before we can assess synergistic effects and extrapolate the additive risk of all mycotoxins present in food. Full article
(This article belongs to the Collection Leading Opinions (Closed))
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2766 KiB  
Article
Silver Nanoparticle-Based Fluorescence-Quenching Lateral Flow Immunoassay for Sensitive Detection of Ochratoxin A in Grape Juice and Wine
by Hu Jiang, Xiangmin Li, Ying Xiong, Ke Pei, Lijuan Nie and Yonghua Xiong
Toxins 2017, 9(3), 83; https://doi.org/10.3390/toxins9030083 - 28 Feb 2017
Cited by 51 | Viewed by 8188
Abstract
A silver nanoparticle (AgNP)-based fluorescence-quenching lateral flow immunoassay with competitive format (cLFIA) was developed for sensitive detection of ochratoxin A (OTA) in grape juice and wine samples in the present study. The Ru(phen) 3 2 + -doped silica nanoparticles (RuNPs) were sprayed on [...] Read more.
A silver nanoparticle (AgNP)-based fluorescence-quenching lateral flow immunoassay with competitive format (cLFIA) was developed for sensitive detection of ochratoxin A (OTA) in grape juice and wine samples in the present study. The Ru(phen) 3 2 + -doped silica nanoparticles (RuNPs) were sprayed on the test and control line zones as background fluorescence signals. The AgNPs were designed as the fluorescence quenchers of RuNPs because they can block the exciting light transferring to the RuNP molecules. The proposed method exhibited high sensitivity for OTA detection, with a detection limit of 0.06 µg/L under optimized conditions. The method also exhibited a good linear range for OTA quantitative analysis from 0.08 µg/L to 5.0 µg/L. The reliability of the fluorescence-quenching cLFIA method was evaluated through analysis of the OTA-spiked red grape wine and juice samples. The average recoveries ranged from 88.0% to 110.0% in red grape wine and from 92.0% to 110.0% in grape juice. Meanwhile, less than a 10% coefficient variation indicated an acceptable precision of the cLFIA method. In summary, the new AgNP-based fluorescence-quenching cLFIA is a simple, rapid, sensitive, and accurate method for quantitative detection of OTA in grape juice and wine or other foodstuffs. Full article
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1829 KiB  
Article
Effects of Electron Beam Irradiation on Zearalenone and Ochratoxin A in Naturally Contaminated Corn and Corn Quality Parameters
by Xiaohu Luo, Lijun Qi, Yuntao Liu, Ren Wang, Dan Yang, Ke Li, Li Wang, Yanan Li, Yuwei Zhang and Zhengxing Chen
Toxins 2017, 9(3), 84; https://doi.org/10.3390/toxins9030084 - 27 Feb 2017
Cited by 35 | Viewed by 5016
Abstract
Zearalenone (ZEN) and ochratoxin A (OTA) are secondary toxic metabolites widely present in grains and grain products. In this study, the effects of electron beam irradiation (EBI) on ZEN and OTA in corn and the quality of irradiated corn were investigated. Results indicated [...] Read more.
Zearalenone (ZEN) and ochratoxin A (OTA) are secondary toxic metabolites widely present in grains and grain products. In this study, the effects of electron beam irradiation (EBI) on ZEN and OTA in corn and the quality of irradiated corn were investigated. Results indicated that EBI significantly affected ZEN and OTA. The degradation rates of ZEN and OTA at 10 kGy in solution were 65.6% and 75.2%, respectively. The initial amounts significantly affected the degradation rate. ZEN and OTA in corn were decreased by the irradiation dose, and their degradation rates at 50 kGy were 71.1% and 67.9%, respectively. ZEN and OTA were more easily degraded in corn kernel than in corn flour. Moisture content (MC) played a vital role in ZEN and OTA degradation. High MC was attributed to high ZEN and OTA degradation. The quality of irradiated corn was evaluated on the basis of irradiation dose. L* value changed, but this change was not significant (p > 0.05). By contrast, a* and b* decreased significantly (p < 0.05) with irradiation dose. The fatty acid value increased significantly. The pasting properties, including peak, trough, breakdown, and final and setback viscosities, were also reduced significantly (p < 0.05) by irradiation. Our study verified that EBI could effectively degrade ZEN and OTA in corn. Irradiation could also affect corn quality. Full article
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