Production of trichothecene and non-trichothecene mycotoxins by Fusarium species isolated from maize in Minnesota

Eighty-two cultures of Fusarium species isolated in 1986 from moldy maize in Minnesota were each cultured on rice for 4 weeks and found to produce the following mycotoxins: F. graminearum isolates, deoxynivalenol (DON, 4–225 g/g), 3-acetyldeoxynivalenol (3-ADON, 2–4g/g), 15-acetyldeoxynivalenol (15-ADON, 1–35 g/g) and zearalenone (ZEA, 5–4350 g/g); F. moniliforme, fusarin C (detectable amounts to 1000 g/g); F. mòniliforme, F. oxysporum, F. proliferatum and F. subglutinans isolates, moniliformin (15–6775 g/g); F. moniliforme, F. proliferatum, and F. subglutinans isolates, fusaric acid (detectable amounts). Other mycotoxins screened for in each rice sample and not detected were T-2 toxin, HT-2 toxin, neosolaniol, T-2 tetraol, nivalenol, fusarenon-X, scirpenols, alpha and beta trans-zearalenols, wortmannin, and fusarochromanone. The rat feeding bioassay indicated that other, unidentified toxins may be present.     

Macrocylic thioether-esters and thioether-thioesters and their palladium, platinum and silver complexes

Preparations by the high dilution method are reported for seven macrocyclic thioether-esters and thioether-thioesters (L1–;L7). Yields in these reactions between thiodiglycolyl dichloride and appropriate ,ω-diols or dithiols range from 10 to 51%. The compounds are characterized by ¹H and ¹³C NMR, IR and high resolution mass spectroscopy. They react with salts of Pd(II), Pt(II) and Ag(I) to form complexes of which MX₂·L2, (M = Pt, X = Cl; M = Pd, X = Cl, Br, I, SCN), [Pd(L2)₂][CF₃SO₃]₂·H₂O and [Ag(L5)₂][CF₃SO₃]·C₂H₅OH have been isolated and characterized by elemental analysis, IR and NMR spectroscopy. NMR spectra indicate reversible dissociation of the ligand occurs in dimethyl sulfoxide solvent for PdCl₂·L2 but not for the Pt analogue. For PtCl₂·L2, spectra indicate that the ligand is undergoing a conformational ‘wag’ about its pair of equivalent sulfurs. These remain bound to the metal while the unique sulfur moves from the apical position of the coordination sphere to a non-coordinated situation. Simultaneously, inversions at the bound sulfurs are occurring.     

Occurrence of trichothecenes, zearalenone and ochratoxin A in cereals and mixed feed from central Lithuania

A total of 92 samples — 23 winter wheat, 12 summer barley, 5 oats and 52 mixed feed — were collected from a state factory in Kaunas, Lithuania and were analysed for the presence of trichothecenes, zearalenone (ZEN) and ochratoxin A (OA) using gas chromatography with electron capture detection and immunoaffinity column/high performance liquid chromatography with fluorescence and UV detections. Deoxynivalenol (DON), nivalenol (NIV), T-2 toxin and HT-2 toxin were detected at concentrations above 10 μg/kg in 68%, 48%, 38% and 8% of cereal samples, respectively, and in 98%, 88%, 12% and 8% of samples of mixed feed for swine and poultry. More than 10 μg/kg of zearalenone and ochratoxin A were found in 58% and 92% of the mixed feed samples, respectively. The highest concentrations of all analysed trichothecenes in Lithuanian mixed feed and cereal grains, with an exception of T-2 toxin in one oat lot and one sample of mixed feed and OA in two mixed feed samples, were lower than those reported as Lithuanian advisory or tolerance limits.     

禾谷镰孢菌高产毒菌株的构建*

为研究禾谷镰孢菌Fusarium graminearum Schw.单端孢霉烯族毒素生物合成基因（产毒基因）在寄主体内的表达,作者构建了带报告基因GUS（(-葡糖苷酸酶基因）的质粒pGUSTRI6P5,并通过对野生型菌株的转化获得禾谷镰孢高产毒菌株。该质粒含有由TRI5（禾谷镰孢单端孢霉二烯合酶基因）启动子（TRI5 Prom）驱动的GUS基因编码区、潮霉素B抗性基因和拟枝孢镰孢F. sporotrichioides的产毒调控基因TRI6（FSTRI6）。用pGUSTRI6P5转化野生型菌株GZ3639后,在含潮霉素 B的培养基上选取抗性菌落,单孢分离获单孢菌株（转化子）。在GYEP（葡萄糖-酵母粉-蛋白胨）液体培养基上,转化子B4-1和B16-1的GUS比活力强,15-AcDON（15-乙酰脱氧雪腐镰刀菌烯醇）产量高,且两者呈正相关（相关系数（r）分别为0.9839和0.9523）。B4-1和B16-1两个转化子可作为研究禾谷镰孢与其寄主相互作用的工具菌株。     

Model studies for isolation of G-quadruplex-forming DNA sequences through a pull-down strategy with macrocyclic polyoxazole

G-quadruplexes (G4s) are non-B DNA structures present in guanine-rich regions of gene regulatory areas, promoters and CpG islands, but their occurrence and functions remain incompletely understood. Thus, methodology to identify G4 sequences is needed. Here, we describe the synthesis of a novel cyclic hepta-oxazole compound, L1Bio-7OTD (1), bearing a biotin affinity-tag as a tool to pull down G4 structures from mixtures of G4-forming and non G4-forming DNA sequences. We confirmed that it could pull down G4s associated with telomeres, bcl-2 gene, and c-kit gene.     

Interacting climate change environmental factors effects on Fusarium langsethiae growth,expression of Tri genes and T-2/HT-2 mycotoxin production on oat-based media and in stored oats

This study examined the effect of climate change (CC) abiotic factors of temperature (20, 25, 30 °C), water activity (a_w; 0.995, 0.98) and CO₂ exposure (400, 1000 ppm) may have on (a) growth, (b) gene expression of biosynthetic toxin genes (Tri5, Tri6, Tri16), and (c) T-2/HT-2 toxins and associated metabolites by Fusarium langsethiae on oat-based media and in stored oats. Lag phases and growth were optimum at 25 °C with freely available water. This was significantly reduced at 30 °C, at 0.98 a_w and 1000 ppm CO₂ exposure. In oat-based media and stored oats, Tri5 gene expression was reduced in all conditions except 30 °C, 0.98 a_w, elevated CO₂ where there was a significant (5.3-fold) increase. The Tri6 and Tri16 genes were upregulated, especially in elevated CO₂ conditions. Toxin production was higher at 25 °C than 30 °C. In stored oats, at 0.98 a_w, elevated CO₂ led to a significant increase (73-fold) increase in T2/HT-2 toxin, especially at 30 °C. Nine T-2 and HT-2 related metabolites were detected, including a new dehydro T-2 toxin (which correlated with T-2 production) and the conjugate, HT-2 toxin, glucuronide. This shows that CC factors may have a significant impact on growth and mycotoxin production by F. langsethiae.     

Exogenous H(2)O(2) and catalase treatments interfere with Tri genes expression in liquid cultures of Fusarium graminearum

Effect of exogenous H(2)O(2) and catalase was tested in liquid cultures of the deoxynivalenol and 15-acetyldeoxynivalenol-producing fungus Fusarium graminearum. Accordingly to previous results, H(2)O(2) supplementation of the culture medium leads to increased toxin production. This study indicates that this event seems to be linked to a general up regulation of genes involved in the deoxynivalenol and 15-acetyldeoxynivalenol biosynthesis pathway, commonly named Tri genes. In catalase-treated cultures, toxin accumulation is reduced, and Tri genes expression is significantly down regulated. Furthermore, kinetics of expression of several Tri genes is proposed in relation to toxin accumulation. Biological meanings of these findings are discussed.     

Effects of four trichothecene mycotoxins on activation marker expression and cell proliferation of human lymphocytes in culture

Four trichothecene mycotoxins – the type A trichothecenes T2-toxin and diacetoxyscirpenol and the type B trichothecenes nivalenol and deoxynivalenol – were studied. The effects of these mycotoxins on the expression of the sequentially expressed activation markers CD69, CD25, and CD71 and on proliferation of human lymphocytes were studied in culture with a duration of up to 72 h.All the examined toxins affected activation marker expression in a similar way. After 6 h, the CD69 expression was lower in exposed cultures compared to controls. After 24 and 48 h of exposure, an increased frequency of cells expressing CD69 was found in exposed cultures, indicating a delay in downregulation of CD69 expression. Stimulation of CD25 expression was observed for doses below the IC₅₀ value, while suppression was found for higher doses. The pattern was different from that detected for CD69 expression, in that an increased expression of CD25 never occurred after exposure to the highest concentration of the toxin, and in that no stimulatory effects were found after 48 h of exposure, indicating that the response was inhibited and not delayed. The effects of toxin exposure on CD71 expression were in many respects similar to the effects on CD25 expression.We conclude that the trichothecene mycotoxins investigated in this study inhibited the cell cycle in a similar way and exert their main antiproliferative action rather early in the cell cycle, before or in conjunction with CD25 expression.