Antiaflatoxigenic effect of fullerene C60 nanoparticles at environmentally plausible concentrations.

Increased interest in fullerene C60 and derivatives in recent years implies an intensification of their environmental spread. Yet, the potential risks for living organisms are largely unknown, including the interaction of C60 with fungal organisms. This may be especially relevant for mycotoxigenic fungi since C60 may both scavenge and produce reactive oxygen species (ROS), and oxidative stress induces mycotoxin production in fungi. Therefore, this study examined effects of environmentally plausible concentrations of C60 (0, 10, 50, and 100 ng/mL) on Aspergillus flavus growth and aflatoxin production in culture media. In addition, ROS-dependent oxidative stress biomarkers-thiobarbituric acid reactive substances (TBARS), reduced and oxidised glutathione ratio, superoxide dismutase isoenzymes, catalase, glutathione peroxidase, and glutathione reductase were determined in mycelia. Nanoparticles of fullerene C60 (nC60) did not exhibit strong antifungal activity against A. flavus. At the same time, nC60 caused an antiaflatoxigenic effect at 10-100 ng/mL, and 50 ng/mL unexpectedly enhanced aflatoxin production. The TBARS content, reduced and oxidised glutathione ratio, and copper, zinc superoxide dismutase activity suggest that 10 ng/mL nC60 exerted antioxidative action and reduced aflatoxin B1 production within fungal cells. Detected prooxidative effects of 50 ng/mL fullerene exceeded cellular defenses and consequently enhanced aflatoxin B1 production. Finally, the results obtained with 100 ng/mL nC60 point to prooxidative effects, but the absence of increase in aflatoxin output may indicate additional, presumably cytotoxic effects of nC60. Thus, a range of rather low levels of nC60 in the environment has a potential to modify aflatoxin production in A. flavus. Due to possible implications, further studies should test these results in environmental conditions.

Fullerol C60(OH)24 nanoparticles and mycotoxigenic fungi: a preliminary investigation into modulation of mycotoxin production.

Increased use of fullerols in various fields and expected increase of their environmental spread impose the necessity for testing fullerol nanoparticles (FNP) effects on microbiota. There is little information available on the interaction of mycotoxigenic fungi and FNP, despite FNP having a great potential of modifying mycotoxin production. Namely, FNP exhibit both ROS-quenching and ROS-producing properties, while oxidative stress stimulates mycotoxin synthesis in the fungi. In order to shed some light on the extent of interaction between FNP and mycotoxigenic fungi, the effects of fullerol C60(OH)24 nanoparticles (10, 100, 1000 ng/mL) on mycelial growth, aflatoxin production and oxidative stress modulation in an aflatoxigenic strain of Aspergillus flavus (NRRL 3251) during 168 h of incubation in a liquid culture medium were examined. FNP slightly reduced mycelial biomass weight, but significantly decreased aflatoxin concentration in media. Lipid peroxide content, superoxide dismutase, catalase and glutathione peroxidase activities suggest that FNP treatments hormetically reduced oxidative stress within fungal cells in turn suppressing aflatoxin production. These findings contribute to the assessment of environmental risk and application potential of fullerols.

Urinary analysis reveals high deoxynivalenol exposure in pregnant women from Croatia.

In this pilot survey the levels of various mycotoxin biomarkers were determined in third trimester pregnant women from eastern Croatia. First void urine samples were collected and analysed using a "dilute and shoot" LC-ESI-MS/MS multi biomarker method. Deoxynivalenol (DON) and its metabolites: deoxynivalenol-15-glucuronide and deoxynivalenol-3-glucuronide were detected in 97.5% of the studied samples, partly at exceptionally high levels, while ochratoxin A was found in 10% of the samples. DON exposure was primarily reflected by the presence of deoxynivalenol-15-glucuronide with a mean concentration of 120 µg L(-1), while free DON was detected with a mean concentration of 18.3 µg L(-1). Several highly contaminated urine samples contained a third DON conjugate, tentatively identified as deoxynivalenol-7-glucuronide by MS/MS scans. The levels of urinary DON and its metabolites measured in this study are the highest ever reported, and 48% of subjects were estimated to exceed the provisional maximum tolerable daily intake (1 µg kg(-1) b.w.).

Blood selenium, glutathione peroxidase activity and antioxidant supplementation of subjects exposed to arsenic via drinking water.

This work investigated serum selenium (Se) and glutathione peroxidase (GPx) levels in 25 Croatian subjects exposed to high levels of As from drinking water (median As level in urine: 620.74µg/g creatinine) and 25 controls (32.98µg/g creatinine). The exposed group had lower (p<0.001) median serum Se and GPx levels (Se: 82.34µg/l vs 59.02µg/l; GPx: 45.99U/g hemoglobin vs 38.38U/g hemoglobin). A subsample of 20 exposed subjects took part in a 2-month antioxidant supplementation trial which increased median GPx activity from 30.71 to 40.98U/g hemoglobin (p=0.041) and reduced total urinary As median from 680.15 to 501.96µg/g creatinine (p=0.051). The effect of selected catalase (-262C>T) and GPx1 (-593C>T) gene polymorphisms was also examined. The low Se status and GPx activity may heighten risk of adverse health effects, especially in genetically predisposed individuals. The outcome of antioxidant treatment indicates modulation of As metabolism and oxidative stress, relevance of which needs further research.

Selenium in placenta predicts birth weight in normal but not intrauterine growth restriction pregnancy.

Placental selenium, lead and cadmium concentrations were determined in a group of pregnancies with birth weight appropriate for gestational age and in a group of intrauterine growth restriction cases. Following adjustment for a number of confounding variables, selenium was found to be a significant predictor of newborn weight only in the group of pregnancies with birth weights appropriate for gestational age. Placental lead and cadmium levels were not associated with birth weight in either group.

Selected markers of cardiovascular disease in a population exposed to arsenic from drinking water.

This study examined prevalence and serum levels of selected markers of cardiovascular disease in 34 subjects from a Croatian rural population exposed to high levels of arsenic (As) from drinking water (611.89±10.06µg/l). The prevalences of overweight and obese subjects in the population were 32% and 35%. Half the subjects had hypertension, 29% had increased fasting serum glucose level and two were diabetic. Median total cholesterol (5.82mmol/l) and triglycerides (2.15mmol/l) were above the desirable margins. The median C-reactive protein level (1.20mg/l) was slightly higher than previously reported for healthy subjects. Serum Hsp70 level was significantly higher in nonsmokers. Total urinary As levels were positively correlated with age-adjusted serum levels of cobalamin. Near significance were also serum total bilirubin, antibodies to Hsp60 and folate. Tentative investigation of risk factors among subjects classified by tumor necrosis factor-α -308G/A and interleukin-6-174G/C gene polymorphisms was also performed. Collectively, the results are in agreement with the hypothesis of As-induced and/or compounded cardiovascular disease.

Contamination of malt barley and wheat by Fusarium graminearum and Fusarium culmorum from the crop years 2001-2003 in eastern Croatia.

This study investigated infection levels with Fusarium graminearum and Fusarium culmorum in malt barley and wheat in eastern Croatia. The contamination was surveyed over three consecutive crop years (2001-2003) on five locations for barley and three wheat cultivating locations. F. graminearum loads reached levels of potentially serious threat for the commercial production of malting raw materials in both cereals (up to 29.1%). On the other hand, the mean percentage of kernels infected with F. culmorum was low to medium (up to 6.1%). The fungal invasions for years and locations were affected by meteorologic and other environmental factors and the pattern seemed to be consistent with species-specific optimal conditions reported by other authors.

High exposure to arsenic from drinking water at several localities in eastern Croatia.

This study investigated the relationship between arsenic concentrations in drinking water in four towns/villages in eastern Croatia and corresponding hair arsenic concentrations of residents. The mean arsenic concentrations in community drinking water samples were 0.14, 37.88, 171.60, and 611.89 microg/l. The corresponding mean concentrations of the element in hair samples of subjects residing in each of the localities were 0.07 (n=11), 0.26 (n=17), 1.74 (n=11), and 4.31 microg/g (n=23). Chronic exposures to arsenic levels estimated in three investigated locations could present a serious health threat to around 3% of Croatian population.