Tomislav Klapec

Daily dietary intake of selenium in eastern Croatia.

Data on the selenium content in the soil and human milk in eastern Croatia indicate a possibility of inadequate Se status of people in the area. In order to determine the daily dietary intake of Se in eastern Croatia, a 7-day duplicate diet study was performed. There were 41 participants (18-53 years old; 14 males and 27 females). Selenium measurement was carried out in a HG-AAS. The average daily Se intake in the area is 27.3 micrograms. A significant difference (P < 0.05) exists between male (32.2 micrograms) and female (24.8 micrograms) participants. Percentage of optimum Se intake shows an inverse association with the increasing age of subjects (r = 0.43; P < 0.01). Relationship between Se intake and intakes of different foods (cereal products, milk and dairy products, meat, fish, etc.) was also examined. The sub-optimal Se intake (RDA is 55 micrograms/day for women and 70 micrograms/day for men) is a reflection of low Se levels in the environment, so supplementation of fertilizers with Se should be considered.

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.).

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.

Urinary ochratoxin A and ochratoxin alpha in pregnant women

This study determined exposure of pregnant women to ochratoxin A (OTA). Forty samples of first-void urine samples from Croatian women in the third trimester of pregnancy were analyzed for OTA and its major metabolite ochratoxin alpha (OTα). The subjects filled a short food frequency questionnaire (FFQ). Analysis was performed by HPLC-FLD following liquid-liquid extraction. All samples were subjected in parallel to enzymatic treatment (β-glucuronidase/aryl sulfatase) to release OTA and OTα from the conjugates. The median urinary levels of OTA and OTα before treatment were 0.02 (range: nd-1.07) ng/mL and 0.16 (nd-1.86) ng/mL; the concentrations after enzyme hydrolysis were 0.02 (nd-1.11) ng/mL and 1.18 (0.11-7.57) ng/mL. While OTα levels increased significantly following enzymatic treatment, evidence for OTA conjugation was weak. The ratio of urinary OTα medians after and before hydrolysis was 1.5 times higher than previously reported for nonpregnant female subjects, possibly indicating upregulated metabolism and/or elimination of the mycotoxin and metabolites in pregnancy. The mean daily dietary OTA intake calculated from FFQs (1.08±0.57 ng/kg body weight) was well below the provisional tolerable daily intake and the greatest contributors to intake were cereal products, fruit juices, chocolate and coffee.

Adolescent acceptance of different foods by obesity status and by sex

This work investigated possible differences in food preference between obese and normal-weight adolescents, as well as between female and male seventh-grade pupils (average age = 13.1 years). Survey of affinities for nine food groups was determined with a facial hedonic scale. The children were divided according to their BMI into normal-weight and obese group, respectively. In this sample of adolescents, we could not find any proof for the theory of increased preference for fattening foods by obese compared to normal-weight people. In fact, our normal-weight group showed greater liking of sweets, meat, and cereals than the obese group. Normal-weight boys also preferred meat and sweets more than normal-weight girls. We consider psychological and social factors (e.g., perception of ideal body shape or social stigmatism of obesity) as the most logical explanations for the observed differences. Further research should clarify whether food preference plays a significant role in the etiology of adolescent obesity.

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.

Distribution of zearalenone in malted barley fractions dependent on Fusarium graminearum growing conditions

Zearalenone (ZON) distribution was measured in main fractions of malted barley, dependent on incubation time (17, 26, 34days), water activity (0.95 and 0.98) and temperature (20°C and 30°C). Malted samples were sterilised and inoculated with Fusarium graminearum. ZON levels were higher (p<0.01) in bran and germ than flour under almost all growing conditions. Incubation at 30°C resulted in generally lower ZON levels in germ and bran, regardless of aw and incubation time. After 34days, ZON levels in flour from samples that were incubated at the higher temperature rose significantly. At 20°C ZON concentration showed a bell-shaped concentration profile with increasing incubation time in bran and germ, whilst ZON levels in flour increased at aw 0.98 and dropped at the lower aw. The results indicate the importance of storage conditions for ZON levels in commercially relevant grain fractions of malted barley and help predict existing mycotoxin levels or manipulate storage conditions to reduce ZON content.

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.

Copper and zinc content in human milk in Croatia

The aim of this paper was to research whether there had been any statistically significant difference in the content of Cu and Zn in human milk depending on the social status of women (refugee and non-refugee), age, number of deliveries, days after delivery, weight gained by nursing women and smoking habits, as well as whether the infants had received sufficient quantities of these elements. The elements were determined by flame atomic absorption spectroscopy. The samples were collected in the Clinical Hospital Osijek and Refugee Centre Nabrde, near Osijek, Eastern Croatia. The Cu in human milk ranged from 0.27 mg/l to 1.35 mg/l, and Zn from 0.62 mg/l to 15.0 mg/l. The mean levels of Cu and Zn for each group, formed according to the results of the questionnaire are presented too. Calculated daily dietary intake of these elements accords with the RDA.

Fat and fatty acids intake of adults in eastern Croatia

The objective of this work was to determine dietary intake of total fat and fatty acids in adults in eastern Croatia. Thirty nine participants (25 women and 14 men) took part in a duplicate diet study. Total fat was extracted from the duplicated portions using a method by Bligh and Dyer, and fatty acids were determined using a GC analysis after methylation with boron trifluoride. Mean fat and fatty acids intakes for all subjects expressed as percentage of daily energy intake (% E) were: 33.4%, 9.9%, 11.8%, 9.0%, and 0.7% for total fat, saturated, monounsaturated, polyunsaturated, and trans fatty acids, respectively. The differences between sexes were not significant when expressed as % E. Also, a very low intake of linolenic acid (0.5% E), and a high linoleic acid/linolenic acid ratio (17.1), were noted. Because of the health implications of such an imbalance, further research on the prevalence of this in general population is warranted. Nevertheless, these results indicate a need for a more aggressive education of the public on health benefits of n-3 fatty acids and on their dietary sources.