Evaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder.

intestinal microbiota is becoming a significant marker that reflects differences between health and disease status also in terms of gut-brain axis communication. Studies show that children with autism spectrum disorder (ASD) often have a mix of gut microbes that is distinct from the neurotypical children. Various assays are being used for microbiota investigation and were considered to be universal. However, newer studies showed that protocol for preparing DNA sequencing libraries is a key factor influencing results of microbiota investigation. The choice of DNA amplification primers seems to be the crucial for the outcome of analysis. In our study, we have tested 3 primer sets to investigate differences in outcome of sequencing analysis of microbiota in children with ASD. We found out that primers detected different portion of bacteria in samples especially at phylum level; significantly higher abundance of Bacteroides and lower Firmicutes were detected using 515f/806r compared to 27f/1492r and 27f*/1495f primers. So, the question is whether a gold standard of Firmicutes/Bacteroidetes ratio is a valuable and reliable universal marker, since two primer sets towards 16S rRNA can provide opposite information. Moreover, significantly higher relative abundance of Proteobacteria was detected using 27f/1492r. The beta diversity of sample groups differed remarkably and so the number of observed bacterial genera.

Specificity of gut microbiota in children with autism spectrum disorder in Slovakia and its correlation with astrocytes activity marker and specific behavioural patterns.

Recent research suggests the involvement of bidirectional gut-brain axis in autism spectrum disorder (ASD). The aim of this study was to establish the role of changed gut microbiota in behavioural and gastrointestinal manifestations, but also in astrocyte activation in children with ASD. Distinct faecal microbiota in children with ASD was found to be more heterogeneous compared to that in neurotypical children. Different bacterial abundance and correlation with behavioural and GI manifestations revealed several bacterial genera possibly important for ASD. Microbial-neuronal cross talk could be accomplished through S100B, released by glial cells, activated by low grade inflammation. More complex studies are required to understand ASD pathogenesis.

Urinary levels of 6-sulphatoxymelatonin and their associations with sleep disorders and behavioural impairments in children with autism spectrum disorder.

BACKGROUND: Autism spectrum disorder (ASD) is a neurodevelopmental condition associated with sleep disturbances that may result from abnormalities in melatonin production. The correlations of melatonin levels with the severity of sleep disorder and/or severity of ASD were reported. OBJECTIVES: To evaluate urinary levels of the melatonin metabolite, 6-sulphatoxymelatonin (aMT6s), in children with ASD, and their associations with sleep abnormalities and behavioural impairments. METHODS: Study involved 77 children with ASD and 84 controls aged 2.5‒15.5 years. Sleep disorders were assessed by Children's Sleep Habits Questionnaire. Morning and afternoon levels of aMT6s were determined by radioimmunoassay method. Urinary creatinine levels were assessed by an enzymatic method. RESULTS: The urinary aMT6s/creatinine values indicate that the night-time melatonin levels are significantly lower in ASD than in controls, but there are no significant differences in the daytime levels. In the ASD group, on average, a 6.8-fold difference between night-time and daytime values of urinary aMT6s/creatinine was found, whereas for the controls a 12.5-fold difference was observed, indicating a lower night-time increase in melatonin levels. In ASD group, the difference in night-time-daytime aMT6s/creatinine value correlated with some types of sleep problems, but not with the severity of ASD. CONCLUSION: The results indicate that in ASD there are differences in the patterns of melatonin secretion that may be associated with sleep impairment (Tab. 4, Fig. 2, Ref. 28).

Erythrocyte deformability in children with autism spectrum disorder: correlation with clinical features.

Biomechanical properties of erythrocytes play an important role in health and disease. Deformability represents intrinsic property of erythrocytes to undergo deformation that is crucial for their passage through the narrow capillaries. The erythrocyte damage can lead to compromised tissue perfusion and consequently play a role in the pathogenesis of various diseases including neurological ones. Data available in databases indicate that erythrocytes in autism spectrum disorder (ASD) are altered. This may affect the clinical symptoms of ASD. The aim of our study was to determine erythrocyte deformability in 54 children with ASD and correlate it with clinical symptoms. We found significant negative correlation between erythrocyte deformability and score in C domain of the Autism Diagnostic Interview-Revised (ADI-R) diagnostic tool describing the measure of restrictive, repetitive, and stereotyped behaviors and interests, mainly observable in C1 and C2, but not in C3 and C4 subdomains. This supports the findings of other authors and suggest that behavioral domain C comprises of more subcategories with different underlying etiology. Our results also indicate that abnormalities in erythrocyte deformability may be involved in ASD pathomechanisms and contribute to its clinical manifestation. Further research is necessary to bring more data and identify erythrocyte deformability as prognostic biomarker in ASD.

Plasma levels of glial cell marker S100B in children with autism.

Autism spectrum disorder (ASD) is a neurodevelopmental condition with increasing incidence. Recent evidences suggest glial cells involvement in autism pathophysiology. S100B is a calcium binding protein, mainly found in astrocytes and therefore used as a marker of their activity. In our study, children with autism had higher plasma concentrations of S100B compared to non-autistic controls. No association of S100B plasma levels with behavioral symptoms (ADI-R and ADOS-2 scales) was found. Plasma S100B concentration significantly correlated with urine serotonin, suggesting their interconnection. Correlation of plasma S100B levels with stool calprotectin concentrations was found, suggesting not only brain astrocytes, but also enteric glial cells may take part in autism pathogenesis. Based on our findings, S100B seems to have a potential to be used as a biomarker of human neurodevelopmental disorders, but more investigations are needed to clarify its exact role in pathomechanism of autism.

Fecal calprotectin levels correlate with main domains of the autism diagnostic interview-revised (ADI-R) in a sample of individuals with autism spectrum disorders from Slovakia.

Autism spectrum disorders (ASD) are neurodevelopmental disorders characterized by impaired social interaction and communication, as well as repetitive behavior and restricted interests. There is convincing evidence that the intestinal inflammation is involved in etiology of ASD. Increased levels of inflammatory markers were shown to be associated with more aberrant behaviors and communication of subjects with ASD. Calprotectin in the feces is produced by activated neutrophils and epithelial cells of the gut mucosa, and its levels reflect local inflammation of the gastrointestinal tract. Concentration of fecal calprotectin was determined by ELISA method in 87 individuals with ASD and 51 controls, of that 29 siblings of children with ASD and 22 non-related controls. In non-relatives significantly lower values of fecal calprotectin were observed than in both subjects with ASD and their siblings. In the group with ASD significant correlations of fecal calprotectin with all domains of the ADI-R diagnostic tool were found: qualitative abnormalities in reciprocal social interaction and communication, restrictive and repetitive patterns of behavior. Results suggest that low grade intestinal inflammation may be one of factors implicated in the pathophysiology of ASD.

[Intestinal microbiota and the brain: multilevel interactions in health and disease]. / Os crevo-mozog: význam interakcií medzi crevnou mikrobiotou a mozgom za fyziologických a patologických situácii.

Commensal bacteria of the digestive tract are separated from the brain by multiple barriers. Despite that bacteria residing in the intestine and the neurons of the brain interact by neural and humoral pathways. Mental processes, such as the stress response, may affect the composition and function of intestinal bacteria via the brain-gut axis. On the other hand, intestinal bacteria can influence the processes in the brain through the gut-brain axis. Disruption of these interactions may be involved in various alterations both in the function of the gastrointestinal tract and the brain function.

Increased plasma levels of the high mobility group box 1 protein (HMGB1) are associated with a higher score of gastrointestinal dysfunction in individuals with autism.

Autism is a disorder of neural development characterized by impairments in communication, social interaction, restricted interests and repetitive behavior. The etiology of autism is poorly understood, the evidence indicates that inflammation may play a key role. In autism a high prevalence of gastrointestinal disturbances is reported, that are linked to a low-grade chronic inflammation of the intestinal mucosa. High mobility group box 1 protein (HMGB1) is an intranuclear protein that can be passively released from necrotic cells or actively secreted under inflammatory conditions as alarmin or late proinflammatory cytokine. The objective of this study was to measure plasma levels of HMGB1 in individuals with autism and to analyze their association with gastrointestinal symptoms. The study involved 31 subjects with low-functioning autistic disorder aged 2-22 years and 16 healthy controls. Plasma HMGB1 levels were significantly higher in individuals with autism than in controls (13.8+/-11.7 ng/ml vs. 7.90+/-4.0 ng/ml, p<0.02). In subjects with plasma HMGB1 levels higher than 11 ng/ml severe forms of GI disorders were more prevalent (83.3 %) than in subjects with lower levels (38.9 %, p<0.04). Results of the study support the involvement of the systemic low-grade inflammation in the pathomechanisms of autism and its possible association with GI symptoms.

Selected biomarkers of age-related diseases in older subjects with different nutrition.

BACKGROUND: The nutritionists introduce on the base of epidemiological and clinical studies that appropriately planned vegetarian diets are healthful, and may provide health benefits in the prevention and treatment of certain diseases. Aging belongs to the main risks of cardiovascular disease. METHODS: Markers of age-related diseases (cardiovascular, metabolic syndrome, diabetes) were assessed in two nutritional groups of older apparently healthy non-obese non-smoking women aged 60-70 years, 45 vegetarians (lacto-ovo-vegetarians and semi-vegetarians) and 38 non-vegetarians (control group on a traditional mixed diet, general population). RESULTS: Vegetarian values of total cholesterol, LDL-cholesterol, triacylglycerols, C-reactive protein, glucose, insulin and insulin resistance are significantly reduced. Non-vegetarian average values of total cholesterol, LDL-cholesterol and C-reactive protein are risk. Vegetarians have a better antioxidative status (significantly increased vitamin C, lipid-standardized vitamine E and beta-carotene plasma concentrations). CONCLUSION: Favourable values of cardiovascular risk markers in older vegetarian women document a beneficial effect of vegetarian nutrition in prevention of this disease as well as the vegetarian diet can be an additional factor in therapy. Vegetarians suffer from mild hyperhomocysteinemia; it is due to the lower vitamin B12 concentration. Vitamin B12 supplements are inevitable for the hyperhomocysteinemia prevention (Tab. 2, Ref. 26).

Vitamin C protective plasma value.

The relationship of plasma concentration and intake of vitamin C was measured in a randomly selected group of 368 apparently healthy adult subjects of two nutritional regimens: traditional mixed diet (general population, n=187) and vegetarians (n=181). The condition of protective plasma concentration over 50 micromol/l (50.3-89.4 micromol/l), the value of which reduces the risk of free radical disease, was found in 87 subjects from the general population group, in whom the average vitamin C intake was 124.2 mg per day in range of 92-181. The recommended dietary allowance for this group in amount of 77 mg of vitamin C daily was calculated from current Slovak recommendations being in amount of 80 mg per day for men and 75 mg for women. Previous epidemiological studies as well as the presented results show that approximately a half of Slovak general population have vitamin C deficient (below 23 micromol/l) or suboptimal (23-50 micromol/l) plasma concentrations with insufficient antioxidative protection. Significantly higher plasma vitamin C concentrations in comparison to non-vegetarians were observed in the vegetarian group. Protective concentrations were noted in 88% of vegetarians vs 46% of non-vegetarians. The significantly reduced values of products of oxidative damage of DNA (DNA breaks with oxidised purines and oxidised pyrimidines), lipids (conjugated dienes of fatty acids, malondialdehyde) and proteins (carbonyls) were found in subjects with plasma vitamin C concentrations being over 50 micromol/l vs. below 50 micromol/l. The data emphasize the role of vitamin C in free radical disease prevention under the condition of protective, antioxidative concentrations. The results of general population group document the need to revise the recommended dietary allowance for vitamin C as well as to change the nutritional habits including regular consumption of fruit and vegetables several times daily (Tab. 3, Ref. 28).

Health benefits and risks of plant proteins.

Plant proteins have a reduced content of essential amino acids in comparison to animal proteins. A significant reduction of limiting amino acids (methionine, lysine, tryptophan) means lower protein synthesis. In subjects with predominant or exclusive consumption of plant food a higher incidence of hypoproteinemia due to significant reduction of methionine and lysine intakes was observed. On the other hand, lower intake of these amino acids provides a preventive effect against cardiovascular disease via cholesterol regulation by an inhibited hepatic phospholipid metabolism. Vegetarians have a significantly higher intake of non-essential amino acids arginine and pyruvigenic amino acids glycine, alanine, serine. When plant protein is high in non-essential amino acids, down-regulation of insulin and up-regulation of glucagon is a logical consequence. The action of glucagon in the liver is mediated by stimulation of adenyl cyclase that raises cyclic-AMP (adenosine-3,5-monophosphate) concentrations. Cyclic-AMP down-regulates the synthesis of a number of enzymes required for de novo lipogenesis and cholesterol synthesis, up-regulates key gluconeogenic enzymes and the LDL receptors and decreases the IGF-1 activity (insulin-like growth factor). Cyclic-AMP thus provides a reduction of atherosclerosis risk factors as well as a retardation of cancer development. A sufficient consumption of plant proteins has the protective effects against chronic degenerative diseases (Tab. 2, Ref. 26).

The effect of selected membrane active substances on erythrocyte deformability.

Aminoguanidine improved the erythrocyte filterability by 4%, pyridoxyliden-aminoguanidine by 11% and pyridoxal by 13% in healthy subjects. In diabetic patients the aminoguanidine effect on erythrocyte filterability was improved by 7%, PAG effect by 9% and pyridoxal effect by 15% in comparison to the control group. The other investigated haematological variables in both groups were within the range of the physiological standard. All of the tested substances demonstrated a mild protective influence on erythrocyte elasticity both in healthy subjects and diabetic patients. Significant elasticity improvement was obtained only by pyridoxal (p<0.01) in patients with diabetes mellitus. (Fig. 4, Ref. 18.).

[Nutritional status in adults on an alternative or traditional diet]. / Nutricný status dospelých na alternatívnom vs tradicnom stravovaní.

BACKGROUND: Plant food lacks vitamin B12, vitamin D and higher n-3 polyunsaturated fatty acids. Essential aminoacids methionine and lysine can be found in significantly lower amounts. On the contrary, the culinary and technologically non-processed plant food and whole-grain products contain essential nutrients in a highly condensed form. The aim of the study was to compare nutritional status of adults on alternative or on traditional diet and sequels of the diet to body metabolism. METHODS AND RESULTS: The group on alternative diet consisted of 89 lacto-ovo-vegetarians (age 38.7 +/- 0.6 years, average duration of vegetarianism 7.8 years). Control group on traditional diet (omnivores, n = 84) was formed as an average sample. Nutritional regime was determined using dietetic questionnaire on the food intake regularity. Vegetarians consume optimal amount of fat (along with recommendations of OVD) with predominance of vegetal lipids. They have low intake of cholesterol (62.8 mg), recommended ratio of saturated fatty acids (SFA), monounsaturated (MUFA), and polyunsaturated (PUFA) 6.5:10.6:8.9 energetic percent. Their ratio of linolic:alpha-linolenic acid (10.4:1) also corresponds with recommendations. In traditional diet, the content of lipids and energy usually exceeds the norm of OVD (by 33% or 19% respectively), cholesterol intake is much higher (512.2 mg, 200 mg is recommended as a maximum). Higher is the amount taken of SFA (11.2 energetic %, recommended 7%), and not sufficient is the intake of alpha-linolenic acid (68% of OVD). People on alternative diet have low plasma levels of risk lipid parameters and significantly higher levels of antisclerotic substances. As a result of significantly higher intake of fruits and vegetables, plant oil, sprouts, seeds, and whole-grain food the plasma levels of antioxidative vitamins are in vegetarians higher then threshold. It reduces the risk of the free-radical disease. On the contrary, vegetarians have deficits in methionine intake, and 15% of them have hypoproteinemia is (0% in omnivores). Low plasma levels of iron and calcium, occurrence of hyposideremia (16% versus 2%) and hypocalcemia (21% versus 8%) corresponds with intake of vegetal absorption inhibitors (fytolic acid, oxalic acid, roughage). Frequently a mild form of hyperhomocysteinemia is found (28% versus 5%), resulting vitamin B12 deficit. CONCLUSIONS: Vegetarian diet is optimal for prevention of free-radical diseases, especially those of the cardiovascular system. It may bring a risk from the point of view of low iron and calcium absorption, low intake of methionine and occurrence of mild forms of hyperhomocysteinemia. In traditional diet, total lipid content should be lowered, amount of vegetable oil with alpha-linolenic acid should be elevated as well as fruit and vegetable consummation. Whole grain food and oily seeds should be included into the daily food.

Correlation of carnitine levels to methionine and lysine intake.

Plasma carnitine levels were measured in two alternative nutrition groups--strict vegetarians (vegans) and lactoovovegetarians (vegetarians consuming limited amounts of animal products such as milk products and eggs). The results were compared to an average sample of probands on mixed nutrition (omnivores). Carnitine levels were correlated with the intake of essential amino acids, methionine and lysine (as substrates of its endogenous synthesis), since the intake of carnitine in food is negligible in the alternative nutrition groups (the highest carnitine content is in meat, lower is in milk products, while fruit, cereals and vegetables contain low or no carnitine at all). An average carnitine level in vegans was significantly reduced with hypocarnitinemia present in 52.9% of probands. Similarly, the intake of methionine and lysine was significantly lower in this group due to the exclusive consumption of plant proteins with reduced content of these amino acids. Carnitine level in lactoovovegetarians was also significantly reduced, but the incidence of values below 30 micromol/l was lower than in vegans representing 17.8% vs. 3.3% in omnivores. Intake of methionine and lysine was also significantly reduced in this group, but still higher compared to vegans (73% of protein intake covered by plant proteins). Significant positive correlation of carnitine levels with methionine and lysine intake in alternative nutrition groups indicates that a significant portion of carnitine requirement is covered by endogenous synthesis. Approximately two thirds of carnitine requirement in omnivores comes from exogenous sources. The results demonstrate the risks of alternative nutrition with respect to the intake of essential amino acids, methionine and lysine, and with respect to the intake and biosynthesis of carnitine.

Homocysteine levels in vegetarians versus omnivores.

Vitamin B(12), folate, and vitamin B(6) are the main determinants of homocysteinemia. The vegan diet provides no vitamin B(12), but also less strict forms of alternative nutrition may suffer from a deficit of this vitamin. The plasma homocysteine level was measured in alternative nutrition groups of adults (lacto- and lactoovovegetarians, n = 62; vegans, n = 32) and compared with the levels in a group consuming traditional diet (n = 59), omnivores). In the group of vegetarians the average homocysteine level is 13.18 vs. 10.19 micromol/l in omnivores; the frequency of hyperhomocysteinemia is 29 vs. 5% in omnivores. In the group of vegans the average homocysteine value is 15.79 micromol/l (53% of the individual values exceeded 15 micromol/l). Omnivores consume the recommended amount of methionine; however, in individuals consuming an alternative diet, the intake of methionine is deficient (assessed by food frequency questionnaire; lower content of methionine in plant proteins). Under conditions of lower methionine availability the remethylation pathway prevails; therefore, vitamin B(12) and folate were evaluated in relation to the homocysteine level. The serum vitamin B(12) levels are significantly lower in the alternative nutrition groups (214.8 pmol/l in vegetarians, 140.1 pmol/l in vegans vs. 344.7 pmol/l in omnivores); a deficit (<179.0 pmol/l) was found in 26% of the vegetarians and in 78% of the vegans vs. 0% in omnivores. The serum folate levels were within the range of reference values in all groups; however, they were significantly lower in omnivores. The results show that the mild hyperhomocysteinemia in alternative nutrition is a consequence of vitamin B(12) deficiency.

Traditional and alternative nutrition--levels of homocysteine and lipid parameters in adults.

Values of homocysteine and lipid parameters were measured in groups of adults consuming alternative nutrition (vegetarians/lactoovo/, vegans) and compared with a group consuming traditional diet (omnivores, general population). Frequency of hyperhomocysteinemia was 53% in the vegans group, 28% in vegetarians vs. 5% in omnivores. In conditions of lower methionine intake (reduced content in plant proteins), the remethylation pathway of homocysteine metabolism prevails and it is vitamin B12 and folate-dependent. The intake of vitamin B12 is equal to zero in vegans; vegetarians consume 124% of the RDA vs. 383% in omnivores. Serum vitamin levels are significantly lower in subjects consuming alternative nutrition with deficiency observed in 24% of vegetarians, 78% of vegans vs. 0% in omnivores. Serum folate levels are within the reference range in all groups. Mild hyperhomocysteinemia in the groups consuming alternative diet is a consequence of vitamin B12 deficiency. Vegetarians and vegans meet the RDA for energy and fat, and have a favourable proportion of saturated, mono- and polyunsaturated fatty acids on total energy intake; the ratio of linoleic/alpha-linolenic acid in their diet corresponds with the recommendations. They have low cholesterol consumption and higher vitamin E and C intake. Optimal fat intake of correct composition is reflected in lower values of atherosclerosis risk factors (cholesterol, LDL-cholesterol, atherogenic index, saturated fatty acids, triacylglycerols), and significantly higher levels of protective substances (linoleic acid, alpha-linolenic acid, HDL-cholesterol, vitamin E, vitamin E/cholesterol, vitamin C). Low lipid risk factors but higher findings of mild hyperhomocysteinemia in vegetarians mean a diminished protective effect of alternative nutrition in cardiovascular disease prevention.

Serum selenium levels in healthy Slovak children and adolescents.

Blood serum selenium levels were measured in 891 healthy children and adolescents (aged 11-18 yr, 450 girls and 441 boys) residing in both rural and urban areas from eight regions of Slovakia. Subjects were divided into four age groups (11-12 y, 13-14 y, 15-16 y, and 17-18 y). Serum selenium concentration was determined by the electrothermal atomic absorption spectrometric method. The mean (+/-SD) serum selenium concentrations were 0.750+/-0.255 micromol/L in girls and 0.773+/-0.235 micromol/L in boys. A large proportion of the individuals (25.7% in girls, 18.1% in boys) exhibited serum selenium levels under 0.57 micromol/L (45 microg/L). An increasing trend of the serum selenium values with age has been observed in both boys (p < 0.01) and girls (p < 0.05). Boys had higher serum selenium levels in the all age groups but the differences were not statistically significant.

[The plasma profile of fatty acids in vegetarians]. / Plazmatický profil mastných kyselín u vegetariánov.

The plasma profile of fatty acids has been observed in 27 vegetarian adults, their age ranging from 20 to 63 years (13 men, 14 women) with the average period of vegetarian dietary pattern being 8.8 years. A third of the men and a half of the women were lacto-vegetarians, the rest of them were lacto-ovo-vegetarians. The contents of saturated and monounsaturated fatty acids insignificantly decreased in vegetarians when compared with non-vegetarians (n = 26, 12 men, 14 women, average group of non-smokers with mixed nutrition). The results of parameters regarded as positive factors in vegetarian diet and effective in prevention of atherosclerosis are as follows: significantly higher values of linoliec acid C 18:2, a significantly higher ratio of linoliec/leic acids C 18:2/C18:1 and insignificantly higher value of linolenic acid C 18:3, together with an unchanged, or moderately decreased contents of poly-unsaturated fatty acids C 20:4 (arachidonic acid) and C 22:6 (n3; docosahexaenoic acid). Further prooxidative-antioxidative parameters were selected in regard to the theoretical anticipation of risk brought about by increased lipoperoxidation in vegetarians (poly-unsaturated fatty acids-substrate of the process) which is significant in the etiology of cardio-vascular and oncologic diseases. Their favourable values (significantly lower plasma contents of conjugated dienes of fatty acids, significantly higher plasma value of vitamin C, Beta-carotene, vitamin E/cholesterol-protection of LDL, vitamin E/triacylglycerols-protection of fatty acids, 3.5 times more significant positive linear correlation of vitamin E/triacylglycerol versus plasma contents of C 18:2) eliminate the risk of increased lipoperoxidation in vegetarians. (Tab. 3, Fig. 1, Ref. 29.)

[Serum levels of vitamins A, C and E in the population of Slovakia]. / Sérové hladiny vitamínov A, C a E u obyvatel'ov Slovenska.

BACKGROUND: Significant forms of vitamin deficiencies are rare in Slovak Republic, however mild hypovitaminoses are still relatively frequent and they have negative consequences on the functional status. Vitamins A, C and E represent an important part of the antioxidant defense system acting against free-radical damage. Imbalance between oxidative stress and antioxidant capacity which leads to free radical damage is considered to be an aetiological factor of ischaemic heart disease and cancer. OBJECTIVES: The study was focused on assessment of serum levels of vitamins A, C and E in the Slovak population older than 35 y and frequency of decreased levels. Relationship between high prevalence of chronic non-communicable diseases and high frequency of suboptimal levels of antioxidant vitamins is discussed. METHODS: Serum levels of vitamin A and E were assessed by the HPLC method, the levels of vitamin C by the colorimetric method. RESULTS: Mean levels of vitamin A were 1.85 mumol/l in both sexes (95% c.i. 1.75-1.89 in males, 1.77-1.92 in females). Mean level of vitamin C was 31.36 mumol/l in males (95% c.i. 30.06-32.72), the value observed in females was significantly higher (43.05 mumol/l, 95% c.i. 41.75-44.40). Mean level of vitamin E observed in males (29.2 mumol/l, 95% c.i. 27.62-30.87) was only slightly, not significantly higher than in females (28.11 mumol/l, 95% c.i. 26.45-29.87). Decreased levels of vitamin E were observed only in a small proportion of the sample, however almost one third of males had decreased level of vitamin C and more than 20% had low level of vitamin A. We observed high prevalence of serum levels of vitamins A, C and E suboptimal from the aspect of risk of coronary heart disease and cancer (vitamin A: males 78%, females 75.5%, vitamin E: 41.9 and 49.6%, respectively, vitamin C: 61.8% and 41.1%). CONCLUSION: Results indicate that sizeable proportion of population has decreased serum levels of vitamins, males especially of vitamin C and females of vitamin A. According to serum levels of vitamin A, C and E high percentage of population can be ranked into the group with increased risk of CHD and cancer. (Tab. 6, Ref. 29.)