Differential modulation of the lipid metabolism as a model for cellular resistance to fumonisin B1-induced cytotoxic effects in vitro.

Differential sensitivity of primary hepatocytes and Chang cells to the cancer promoter fumonisin B1 (FB1)-induced cytotoxic effects were investigated in relation to changes in membrane lipid distribution. In contrast to primary hepatocytes, Chang cells were resistant to FB1-induced cytotoxic effects. This was associated with a high cholesterol (Chol) and sphingomyelin (SM) and low phosphatidylcholine (PC) content, resulting in a significant (P<0.05) decrease in phosphatidylethanolamine (PE)/PC ratio, increased Chol/total phosphoglyceride (TPG) ratios and low total polyunsaturated fatty acids (PUFA) content in PC and PE, suggesting a more rigid membrane structure. High levels of C18:1 and reduced polyunsaturated fatty acid (PUFA) levels are likely to provide selective resistance to FB1-induced oxidative stress. FB1-associated lipid changes included decreases in SM and Chol, increases in sphinganine (Sa) and PE with the increases in key saturated, monounsaturated, and PUFAs in PE as key role players in the differential responses to FB1-induced cell growth responses in cells.

Fusarium species and fumonisins in subsistence maize in the former Transkei region, South Africa: a multi-year study in rural villages.

Fumonisin occurrence was investigated in subsistence maize in four rural villages in each of Mbizana and Centane areas, South Africa. Samples (total 211) were analysed morphologically for Fusarium species and by high performance liquid chromatography for fumonisins. The mean incidence levels of Fusarium verticillioides in Centane good maize were 16% for both 1997 and 2000, but increased to 32% in 2003, whereas Mbizana good maize contained levels of 17% and 11% (2000 and 2003 seasons, respectively). The mean total fumonisin level in good maize in Centane for 1997 and 2000 was 575 and 975 µg/kg and 2150 µg/kg in 2003. In Mbizana, the mean total fumonisin level in good maize for 2000 was 950 µg/kg, but decreased to 610 µg/kg in 2003. The 2003 drought conditions led to a substantial increase in fumonisin levels in dry subhumid Centane, compared to humid subtropical Mbizana. This study emphasises the seasonal fluctuation in fumonisin levels.

Single-kernel analysis of fumonisins and other fungal metabolites in maize from South African subsistence farmers.

Fumonisins are important Fusarium mycotoxins mainly found in maize and derived products. This study analysed maize from five subsistence farmers in the former Transkei region of South Africa. Farmers had sorted kernels into good and mouldy quality. A total of 400 kernels from 10 batches were analysed; of these 100 were visually characterised as uninfected and 300 as infected. Of the 400 kernels, 15% were contaminated with 1.84-1428 mg kg(-1) fumonisins, and 4% (n=15) had a fumonisin content above 100 mg kg(-1). None of the visually uninfected maize had detectable amounts of fumonisins. The total fumonisin concentration was 0.28-1.1 mg kg(-1) for good-quality batches and 0.03-6.2 mg kg(-1) for mouldy-quality batches. The high fumonisin content in the batches was apparently caused by a small number (4%) of highly contaminated kernels, and removal of these reduced the average fumonisin content by 71%. Of the 400 kernels, 80 were screened for 186 microbial metabolites by liquid chromatography-tandem mass spectrometry, detecting 17 other fungal metabolites, including fusaric acid, equisetin, fusaproliferin, beauvericin, cyclosporins, agroclavine, chanoclavine, rugulosin and emodin. Fusaric acid in samples without fumonisins indicated the possibility of using non-toxinogenic Fusaria as biocontrol agents to reduce fumonisin exposure, as done for Aspergillus flavus. This is the first report of mycotoxin profiling in single naturally infected maize kernels.

Occurrence of the C-series fumonisins in maize from the former Transkei region of South Africa.

Fumonisins are a group of structurally related mycotoxins produced mainly in maize by Fusarium verticillioides and F. proliferatum. The most abundant naturally occurring analogue is fumonisin B(1) (FB(1)), with lesser amounts of fumonisin B(2) (FB(2)) and fumonisin B(3) (FB(3)) occurring. The C-series fumonisins (FCs) are structurally analogous to the B-series but lack the C-1 methyl group. Good and mouldy subsistence-grown maize samples were collected from the Centane and Bizana districts in the former Transkei region of South Africa. After extraction with methanol/water and clean-up on strong anion exchange solid phase extraction cartridges, FB(1), FB(2), FB(3), FC(1), FC(3) and FC(4) were determined by reversed-phase LC-MS/MS using positive ion electrospray ionisation. FB(1) levels in both good and mouldy maize from Centane (means (±SD) 2.75 ± 2.24 and 23.4 ± 12.5 mg kg(-1), respectively) were higher than the corresponding levels in maize samples from Bizana (means 0.056 ± 0.157 and 3.71 ± 5.01 mg kg(-1), respectively). Similarly, FC(1) levels in both good and mouldy maize from Centane (means 0.107 ± 0.099 and 0.814 ± 0.391 mg kg(-1), respectively) were higher than in Bizana, where FC(1) was detected in only one (0.018 mg kg(-1)) of 19 good maize samples and occurred in mouldy maize with a mean of 0.102 ± 0.135 mg kg(-1). A significant correlation (r=0.982, p<0.01) was observed between FB(1) and FC(1) levels in all samples, with FC(1) levels at 3.3% of the corresponding FB(1) levels. FC(4) levels were similar to FC(1), whereas only low amounts of FC(3) were detected.

Simple intervention method to reduce fumonisin exposure in a subsistence maize-farming community in South Africa.

In the Centane magisterial area of South Africa, high rates of oesophageal cancer have been associated with home-grown maize contaminated with fumonisins. The aim of this study was to implement a simple intervention method to reduce fumonisin exposure in a subsistence-farming community. The hand-sorting and washing procedures, based on traditional maize-based food preparation practices, were previously customised under laboratory-controlled conditions. Home-grown maize and maize-based porridge collected at baseline were analysed for fumonisin B(1), B(2) and B(3). The geometric mean (95% confidence interval) of fumonisin contamination in the home-grown maize at baseline was 1.67 (1.21-2.32) mg kg(-1) and 1.24 (0.75-2.04) mg kg(-1) (dry weight) in the porridge. Fumonisin exposure was based on individual stiff porridge consumption and the specific fumonisin levels in the porridge (dry weight) consumed. Porridge (dry weight) consumption at baseline was 0.34 kg day(-1) and fumonisin exposure was 6.73 (3.90-11.6) µg kg(-1) body weight day(-1). Female participants (n = 22) were trained to recognise and remove visibly infected/damaged kernels and to wash the remaining maize kernels. The discarded kernels represented 3.9% by weight and the fumonisins varied from 17.1 to 76.9 mg kg(-1). The customised hand-sorting and washing procedures reduced fumonisin contamination in the maize and porridge by 84 and 65%, respectively. The intervention reduced fumonisin exposure by 62% to 2.55 (1.94-3.35) µg kg(-1) body weight day(-1). This simple intervention method has the potential to improve food safety and health in subsistence-farming communities consuming fumonisin-contaminated maize as their staple diet.

Dietary fumonisin exposure in a rural population of South Africa.

A validated culturally specific dietary assessment method was used to determine the habitual maize intakes of black Xhosa-speaking Africans living in the Centane region of the Eastern Cape Province to assess their exposure to the carcinogenic fumonisin mycotoxins. The mean total dry weight maize intakes of home-grown, commercial or combined (both maize sources) were 474, 344, 462 g day(-1), respectively. When considering the total mean levels of fumonisin in home-grown maize (1142 microg kg(-1)) and commercial maize (222 microg kg(-1)), the probable daily intakes (PDI's), expressed as microg kg(-1) body weight day(-1) were 12.1 (95%CI: 0.3-4926.5) and 1.3 (95%CI: 1.0-1.8) for men and 6.7 (95%CI: 1.0-457.8) and 1.1 (95%CI: 0.9-1.3) for women, consuming home-grown and commercial maize, respectively. Based on the different maize-based beer drinking frequencies the PDI's varied between 6.9 and 12.0 microg kg(-1)/drinking event. Depending on the maize intake patterns an exposure "window" exists where fumonisin exposure is below the recommended group provisional maximum tolerable daily intake (PMTDI) for fumonisins of 2 microg kg(-1)bw day(-1). The assessment of fumonisin exposure and development of preventative strategies depend, not only the accurate determination of total fumonisin levels in maize, but also on the distinct dietary patterns of a specific population.

Individual fumonisin exposure and sphingoid base levels in rural populations consuming maize in South Africa.

Low and high oesophageal cancer incidence areas of the former Transkei region of South Africa have been associated with corresponding low and high levels of fumonisin contaminated home-grown maize. This is the first study in South Africa assessing fumonisin B (FB) mycotoxin exposure by quantifying individual maize consumption with weighed food records and FB levels from maize in each participant's household and concurrently evaluating sphinganine (Sa), sphingosine (So) and Sa/So ratios in plasma and urine of these participants as possible biomarkers of FB exposure. The high consumption of maize in Bizana (n=36) and Centane (n=30) of 0.41+/-0.21 and 0.39+/-0.19 kg/day, respectively, confirms the reliance on maize as the dietary staple. Mean total FB (FB(1)+FB(2)+FB(3)) levels in home-grown maize were 0.495+0.880 and 0.665+0.660 mg/kg in Bizana and Centane, respectively. Mean fumonisin exposure based on individual consumption was 3.9+/-7.3 and 4.1+/-7.6 microg/kg body weight/day, respectively, for Bizana and Centane. The mean combined sphinganine/sphingosine ratios in Bizana and Centane were similar and ranged from 0.10-0.55 in plasma (n=41) and urine (n=62). There was no association between sphingoid base levels and/or Sa/So ratios in the plasma and urine and individual fumonisin exposure, negating the sphingoid bases as potential biomarkers of fumonisin exposure in humans.

Occurrence of fumonisins in maize imported into Iran during 2001-2002.

Fumonisins, fungal toxins found primarily in maize and produced by various Fusarium species, have been shown to cause a variety of significant adverse health effects in livestock and experimental animals, and are probable human carcinogens. Thirty-three maize samples were collected at ports from bulk shipments, which were imported into Iran from six countries during 2001-2002, and analysed by HPLC for the most abundant of the naturally occurring fumonisin analogues, namely fumonisins B1 (FB1), B2 (FB2) and B3 (FB3). Of the 33 samples, 21 (64%) were found to contain FB1 (58-512 µg/kg) at levels above 10 µg/kg. The frequency of FB1 found in maize samples imported from Uruguay and Canada was 75%, followed by China and Argentina (67%), USA (60%), and Brazil (50%). The average FB1 level was 266 and 169 µg/kg for positive and all samples, respectively. Medians were 250 and 146 µg/kg for positive and all samples, respectively. FB2 levels ranged from not detected (<10 µg/kg) to 53 µg/kg, whereas no sample had an FB3 level above the detection level (10 µg/kg). This is the first report of fumonisin contamination of imported maize in Iran. Although, the level of all detected fumonisins were below the Iranian and FDA tolerance levels for foods and feeds, It is necessary to maintain the strict rules to ensure continued safety of imported maize.

Sphingoid base levels in humans consuming fumonisin-contaminated maize in rural areas of the former Transkei, South Africa: a cross-sectional study.

High incidences of oesophageal cancer are associated with the consumption of subsistence-grown maize by rural populations in the former Transkei region of Eastern Cape Province, South Africa. This cross-sectional study was conducted in the north-eastern magisterial area of Bizana (a previously low oesophageal cancer incidence area) and the south-eastern area of Centane (a previously high incidence area). Plasma and urine samples of male and female participants were analysed for the sphingoid bases, sphinganine and sphingosine. Good home-grown and visibly mouldy maize samples, collected from the households of the participants, were analysed for fumonisin B(1), B(2) and B(3). Plasma sphinganine/sphingosine ratios in males and females were significantly lower (p < 0.05) due to lower sphinganine levels in Bizana compared to Centane. In contrast, the urinary female and combined (males + females) sphinganine/sphingosine ratios were significantly higher (p < 0.05) in Bizana due to the significantly lower (p < 0.05) urinary sphingosine levels. Interestingly, urinary sphingoid base levels were significantly lower (p < 0.05) in males than females within each area. Based on the mean total fumonisin levels in good maize, the estimated mean probable daily intake (PDI) was 5.8 microg kg(-1) body weight day(-1) in Bizana during 2000 and 4.4 and 6.7 5.8 microg kg(-1) body weight day(-1) in Centane during 1997 and 2000, respectively, exceeding the maximum tolerable daily intake proposed by JECFA. However, there was no significant difference in the mean total fumonisin levels in the maize between the magisterial areas. The observed differences in plasma and urinary sphingoid base levels could not be ascribed as a biomarker of fumonisin exposure and further studies at an individual level are required.

Biomarkers of exposure to fumonisin mycotoxins: a review.

The investigation of adverse health effects associated with fungal mycotoxins requires the measurement of human exposure. Most frequently, this exposure is estimated from contamination levels of raw foodstuffs, which are the primary source of toxin exposure, and data on food consumption patterns. However, variations in food preparation methods, food intake, contamination level, intestinal absorption, toxin distribution and excretion lead to individual variations in toxin exposure that are more readily measured with a biomarker. Fumonisin biomarkers have been sought in the measurement of levels of the toxin in physiological samples such as serum, urine, faeces, hair and nails. However, due to the low bioavailability of fumonisin, these samples pose a variety of analytical challenges and also still require validation as biomarkers. The most widely researched fumonisin biomarkers have been those related to the disruption of de novo sphingolipid biosynthesis, namely elevated levels of the sphingoid base, sphinganine, or of its ratio with sphingosine. Elevation of these parameters in humans would potentially provide a biomarker of biochemical effect. A number of investigations into the possible elevation of sphinganine (or its ratio with sphingosine) in human blood and urine have generally failed to correlate with estimates of fumonisin exposure. The sphingoid bases occur naturally in human blood and urine such that their levels have normal ranges, which can be influenced by dietary factors other than fumonisin ingestion. The lower exposures from human diets, as compared with doses in experimental animals, have made detection of changes in these sphingoid biomarkers problematic.

Exposure assessment for fumonisins in the former Transkei region of South Africa.

The fumonisins are mycotoxins produced mainly by Fusarium verticillioides and F. proliferatum in maize, the predominant cereal staple for subsistence farming communities in southern Africa. In order to assess exposure to these mycotoxins in the Bizana (now known as Mbizana) and Centane magisterial areas of the former Transkei region of the Eastern Cape Province of South Africa, the actual maize consumption by different age groups in these communities was measured. In the groups 1-9 years (n = 215) and 10-17 (n = 240) years, mean consumption (+/-standard error) was 246 +/- 10.8 and 368 +/- 10.3 g per person day(-1), respectively, with no significant difference (p > 0.05) between the magisterial areas. For adults (18-65 years) mean maize consumption in Bizana (n = 229) and Centane (n = 178) were significantly different (p < 0.05) at 379 +/- 10.5 and 456 +/- 11.9 g per person day(-1), respectively. An exposure assessment was performed by combining the maize consumption distribution with previously determined levels of total fumonisin (fumonisins B(1) and B(2) combined) contamination in home-grown maize in these two areas. Assuming an individual adult body weight of 60 kg, fumonisin exposure in Bizana, an area of relatively low oesophageal cancer incidence, was 3.43 +/- 0.15 microg kg(-1) body weight day(-1), which was significantly lower (p < 0.05) than that in Centane (8.67 +/- 0.18 microg kg(-1) body weight day(-1)), an area of high oesophageal cancer incidence. Mean fumonisin exposures in all age groups in both Bizana and Centane were above the provisional maximum tolerable daily intake (PMTDI) of 2 microg kg(-1) body weight day(-1) set by the Joint FAO/WHO Expert Committee on Food Additives.

Human dietary exposure to fumonisin B1 from Iranian maize harvested during 1998-2000.

Fumonisin B(1) (FB(1)) is the most abundant of the fumonisin mycotoxins, mainly produced in maize by F. verticillioides and F. proliferatum. A previous study on the FB1 contamination of maize harvested in Mazandaran and Isfahan Provinces of Iran in 1998 and 1999 demonstrated contamination in both provinces. This present study was undertaken to further investigate the variation in levels of contamination and to estimate possible levels of human exposure to fumonisins in Iran. The mean level of FB(1) in 49 visually healthy maize samples collected from Mazandaran Province during 2000 was 6.14 mg/kg, which is higher than that found during 1998 and 1999 (2.27 and 3.18 mg/kg, respectively). Although these levels are higher than the Iranian legislative limits for fumonisins in maize intended for humans, the relatively low estimated consumption of maize in Iran (3.3 g/person/day) implies that average exposures (0.011 and 0.215 microg/kg body weight/day in Isfahan and Mazandaran, respectively) are within the provisional maximum tolerable daily intake of 2 microg/kg body weight/day set by the Joint FAO/WHO Expert Committee on Food Additives. Nevertheless, certain sections of the population who may consume higher amounts of maize or who may replace all or some of their consumption of other cereals with maize, could well exceed this limit.

Episodic cervical dystonia associated with gastro-oesophageal reflux. A case of adult-onset Sandifer syndrome.

Sandifer syndrome is a dystonic movement disorder described in children with severe gastro-oesophageal reflux. We now report a patient who had the features of Sandifer syndrome first developing in adult life. Onset of dystonic episodes followed closely the occurrence of a Bell's palsy, while symptoms of peptic oesophagitis had been present for several months beforehand. Successful symptomatic treatment of gastro-oesophageal reflux was accompanied by cessation of the dystonic episodes. Possible pathophysiological mechanisms of the abnormal movements in Sandifer syndrome are discussed.

The effect of a single gavage dose of fumonisin B(2) on the sphinganine and sphingosine concentrations in vervet monkeys.

The disruption in sphinganine (Sa) and sphingosine (So) concentrations in plasma and urine of vervet monkeys (Cercopithecus aethiops) was measured following a single gavage dose of either 1 (low dose) or 10 mg (high dose) fumonisin B(2) (FB(2))/kg body weight. Blood and urine were collected over a 51-day period. In the low-dose monkeys, none of the parameters measured increased significantly above the control values. In the high-dose monkeys the plasma Sa/So ratios were significantly increased (P< 0.05) above the corresponding control ratios after 3 days and continued to be significantly raised for another 27 days, whereafter the ratios declined to control values after 51 days. The plasma aspartate transaminase (AST) activities increased significantly above their control values from day 5 to day 23 and the gamma-glutamyl transferase (GGT) activities from day 7 until the end of the study period. The plasma cholesterol concentration and alanine transaminase (ALT) and lactate dehydrogenase (LDH) activities increased transiently, but not significantly, and returned to control values within the study period. The urinary Sa/So ratio, plasma creatinine and urea values in both groups of monkeys did not increase above the control values. This study demonstrated that a single large dose of FB(2) can cause transient disruption of sphingoid metabolism in vervet monkeys.

The effect of repeated gavage doses of fumonisin B1 on the sphinganine and sphingosine levels in vervet monkeys.

Fumonisins occur widely around the world in maize products intended for human and animal consumption posing a health hazard. Direct measurement of fumonisins in physiological fluids as a biomarker of exposure is not practicable, however fumonisins disrupt sphingoid metabolism by inhibition of sphinganine N-acyltransferase. In this study the disruption in sphinganine (Sa) and sphingosine (So) levels in plasma and urine was measured in vervet monkeys (Cercopithecus aethiops) dosed with repeated gavages of 1mg fumonisin B1 (FB1)/kg body weight three times/week continuously over a 51-day period. The plasma Sa/So ratio reached a maximum (mean of 4.3) after 30 days in the dosed monkeys with a three-fold increase above the ratio of the control monkeys and then declined slowly to double the value in controls after 51 days. The lack of a clear elevation in urinary Sa/So ratios after 51 days of multiple exposure in the dosed monkeys indicates that the plasma ratio is more sensitive than urinary changes in monkeys. This is confirmed by the plasma levels of liver function enzymes of which aspartate transaminase, glutamyl-transferase and lactate dehydrogenase were increased in the dosed monkeys, while the plasma indicators of renal function were not increased above the levels in the control monkeys. Thus repeated low doses of FB1 can cause sustained disruption of sphingoid metabolism.

The effect of a single gavage dose of fumonisin B(1) on the sphinganine and sphingosine levels in vervet monkeys.

This is the first report of sphinganine (Sa) and sphingosine (So) levels determined in serum and urine of vervet monkeys (Cercopithecus aethiops) dosed with pure fumonisin B(1) (FB(1)). Initially, experimental vervet monkeys were given a single gavage dose of either 1 or 10 mg FB(1) /kg body weight. Blood and urine were sampled daily and on day seven the monkeys were terminated and the kidneys and livers harvested. In a subsequent experiment, other vervet monkeys were similarly dosed and blood and urine samples were collected over a 50-day period. In the high-dose monkeys the serum Sa/So ratio, as well as levels of serum cholesterol and liver function enzymes, increased during the first week after dosing and remained elevated for several weeks thereafter. The urinary Sa/So ratio and the serum renal function indicators showed a more rapid response and a correspondingly more rapid return to pre-dosing levels. In the low-dose monkeys serum Sa and the Sa/So ratio were the only parameters to increase above the control levels. The Sa/So ratio in liver and kidney tissue showed an elevation over controls in a dose-dependent manner. The serum Sa/So ratio was exclusively elevated above the control levels in the low- and high-dose monkeys and seems more relevant as a marker for fumonisin exposure than any of the other indicators.

Multiple infarcts in a patient with cerebral phaeohyphomycosis: CT and MRI.

Phaeohyphomycosis is an uncommon disorder caused by a variety of saprophytic fungi having distinctive morphologic features. Central nervous system infection typically occurs in the absence of predisposing factors and usually manifest symptoms and signs of abscess formation. We describe an otherwise healthy young man whose presentation with cerebral phaeohyphomycosis was subacute meningitis and stroke. Neuroimaging studies revealed multiple parenchymal lesions having the characteristics of recent infarcts; several vascular territories were involved. The nature of these lesions was confirmed histologically at autopsy. To our knowledge, such radiologic appearances have not previously been reported in this condition.

Expertise and performance on the emotional Stroop task.

40 undergraduate students participated in an emotional Stroop task to test the hypothesis that individuals with high interest in and formal knowledge of interpersonal relationships will react with significantly longer latencies to social-related threat words than individuals without such an interest or formal knowledge. The results did not support the hypothesis; however, an association between interference scores and order of card presentation was found.

Sphinganine/sphingosine ratio in plasma and urine as a possible biomarker for fumonisin exposure in humans in rural areas of Africa.

This study was conducted in the Transkei region of the Eastern Cape and KwaZulu-Natal province, South Africa and in the Bomet district, western Kenya. The sphinganine (Sa)/sphingosine (So) ratios in the plasma and urine of male and female volunteers consuming a staple diet of home-grown maize in Transkei, were 0.34 +/- 0.36 (mean +/- standard deviation) (n = 154) and 0.41 +/- 0.72 (n = 153), respectively and in plasma samples from KwaZulu-Natal it was 0.44 +/- 0.23 (n = 26). In Kenya, the ratios in plasma and urine were 0.28 +/- 0.07 (n = 29) and 0.34 +/- 0.20 (n = 27), respectively. Mean total fumonisin level in home-grown maize, randomly collected in Transkei from the same region where the human volunteers lived, was 580 ng/g (n = 40), as compared to the KwaZulu-Natal province, where no fumonisin (n = 17) were detected (< 10 ng/g) in the home-grown maize. In Kenya, only one of seven samples was contaminated with 60 ng/g fumonisins. No significant differences were found in the Sa/So ratios between males and females within the regions nor between the different regions (P > 0.05). It is possible that the ratio is not sensitive enough to act as a biomarker for fumonisin exposure in humans at these levels of contamination in maize. This is the first report on Sa/So ratios determined in rural populations in Africa consuming home-grown maize as their staple diet.

Liquid chromatographic determination of the sphinganine/sphingosine ratio in serum.

The fumonisin mycotoxins, which are worldwide contaminants of corn, inhibit de novo sphingolipid biosynthesis leading to elevation in the ratio of the sphingoid bases, sphinganine and sphingosine, in the serum of animals exposed to fumonisins. A new HPLC method for the determination of the ratio of these bases in serum has been developed involving lipid extraction, clean-up on a silica minicolumn and alkaline hydrolysis prior to precolumn o-phthaldialdehyde derivatisation and HPLC separation and quantification by fluorescence detection. Based on serum from both normal and fumonisin-exposed vervet monkeys, the method was shown to be reproducible (R.S.D.<10%).