Response of hydroponically grown head lettuce on residual monomer from polyacrylamide.

The aim was to assess acrylamide monomer (AMD) uptake by hydroponically grown lettuce. Lettuce was cultivated by applying plant tissue testing in a recycled system by the use of nutrient solutions prepared with two water-soluble flocculants F3 and F4 containing 176 and 763 mg kg(-1) of AMD, respectively. The effects on growth, fresh weight and plant leaf quality were evaluated by comparing these treatments and one control standard nutrient solution typically recommended for lettuce hydroponic cultivation. To assess the nutritional status of lettuce, samples were collected and lyophilised before determination of the selected micro- and macro-element contents. An HPLC with photodiode array detector method was applied to determine AMD in both selected flocculants and dried plant samples. Results show that lettuces cultivated under the conditions described above absorb AMD from nutrient solutions into their leaves. The AMD presence in recycled nutrient solutions has a negative influence on the growth of lettuce, reducing their average fresh weight and average number of leaves. The study confirmed that the problem of AMD mobility and its accumulation risk in plants should to be an important topic with respect to safe polyacrylamide (PAM) handling in the agro food area.

Ochratoxin A and citrinin production by Penicillium verrucosum on cereal solid substrates.

Cereals are among the most vulnerable substrates to mycotoxin contamination. One of the major fungal species colonising the ecosystem of stored grain is Penicillium verrucosum because of its ability to produce ochratoxin A (OTA). Knowledge of favourable environmental conditions promoting fungal growth and mycotoxin biosynthesis seems to be a crucial step towards preventing mycotoxin formation in stored grain. The aim of the study was to examine the effect of temperature and different cereal substrates (wheat, triticale, rye, barley, maize, rice) on fungal growth and OTA and citrinin (CIT) production by a mycotoxin-producing P. verrucosum strain at one moisture content. The correlation between fungal growth and mycotoxin biosynthesis was also investigated. For this purpose, cereal substrates moistened, autoclaved and inoculated with P. verrucosum spores were stored at 10°C, 20°C and 30°C for 40 days. After the storage period, the ergosterol (ERG) content as a fungal growth biomarker and OTA and CIT levels were determined using HPLC. Fungal growth was observed on all cereals at each temperature. Maximum ERG levels were found at 30°C and the values ranged from 401 to 3280 mg kg(-1) of grain. Among tested cereals, the highest ergosterol content was found on rice at all applied temperatures. Accumulation of OTA and CIT was detected in samples stored at 20°C and 30°C, while optimal conditions for the production of both mycotoxins were observed at 20°C. The maximum concentration of OTA (31.9 ± 4.3 (SD) µg kg(-1)) and CIT (74.8 ± 5.7 µg kg(-1)) were found on rice. The ability to biosynthesise OTA and CIT at 10°C was not detected. Analysis of variance showed that single factors, temperature and cereal matrices and their interactions had significant effects on ERG, OTA and CIT levels. Optimal conditions for fungus growth did not coincide with the ones for mycotoxin formation.

Fusarium temperatum as a New Species Causing Ear Rot on Maize in Poland.

Diseases caused by Fusarium spp. are probably the most important factors affecting the yield and grain quality of maize (Zea mays L.) in Poland. While Fusarium stalk rot problems were reported to be stable in all regions, ear rot tends to increase. This tendency probably results from warmer climate and conservation tillage techniques combined with maize- and wheat-dominated rotation systems increasingly practiced in these regions. Maize kernel samples were harvested from trials conducted with hybrids moderately resistant and susceptible to ear rot in three locations in 2011 (12 samples) and in seven locations in 2012 (20 samples) from regions of Poland where maize for seed production is grown for many years. For each sample, 56 kernels (1,792 total) were subjected to the investigation of fungal colonization as following: surface-disinfected in alcohol (15 sec), rinsed with distilled water, dried, placed on the water agar in petri plates, incubated 7 to 12 days at 22°C, and sub-cultured using the single spore technique on SNA. Pure cultures were grown on SNA at 22°C for 10 days to produce macroconidia of uniform size and form, as well as on PDA (1). The presence of Fusarium spp. was determined based on the SCAR-PCR markers and the translation elongation factor (tef-1α) sequence analyses (4). All samples were contaminated by Fusarium spp. at level ranging from 17.3 to 40.5% of kernels (444 strains were isolated). F. verticillioides was the most frequent species. The frequencies of F. temperatum (a recently described species closely related to F. subglutinans [3]) ranged from 2.3 to 54.3%, F. subglutinans from 0 to 37.9%, F. verticillioides 41.0 to 90.0%, and F. proliferatum from 0 to 11.5% of the total Fusarium strain number. After extraction and purification, Fusarium mycotoxins were detected using HPLC method. The majority of maize kernel samples contained fumonisins B1 to B3. Zearalenone and deoxynivalenol were not detected or their levels were below limit detections. Aggressiveness of 10 representative isolates (3 each of F. temperatum, F. verticillioides, and F. subglutinans, and 1 F. proliferatum) was evaluated under field conditions after kernel inoculation. Susceptible and moderate resistant inbreed lines were used. The experimental design consisted of a factorial arrangement with two factors and three replicates (7 plants per replicate). To produce inoculum, isolates were grown on a liquid SNA medium. After 2 weeks, cultures were filtered through cheesecloth and conidial concentrations were adjusted to ~106 spores ml-1. Inoculation of each ear was conducted 10 to 12 days after silking time using 1.5 ml of spore suspension. Control plants were inoculated with distilled water. Visual evaluation of ear rot severity was scored during harvesting time as the percentage of the ear area covering by mycelium (2). Differences of aggressiveness between genotypes, Fusarium spp., and isolates were statistically significant; ear area covered by mycelium ranged from 3.57 to 26.95% for F. temperatum, from 10.10 to 41.39% for F. subglutinans, from 5.90 to 14.33% for F. verticillioides, and from 9.57 to 17.14% for F. proliferatum. To our knowledge, this is the first report of F. temperatum causing ear rot of maize in Poland. References: (1) J. F. Leslie et al. Page 388 in: The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, 2006. (2) L. M. Reid et al. Can. J. Plant Pathol. 19:185, 1997. (3) J. Scauflaire et al. Mycologia 103:586, 2011. (4) L. Stepien et al. J. Appl. Genet. 54:367, 2013.

Estimation of dietary intake and patterns of polyphenol consumption in Polish adult population.

PURPOSE: Polyphenols play an important role in the prevention of degenerative diseases, particularly cardiovascular diseases and cancers. The purpose was to estimate dietary polyphenol intake in Polish adults, using own database of food polyphenols, and to establish main dietary sources of polyphenols. MATERIAL AND METHODS: Polish men and women (6661) aged 20-74 years were randomly selected from the Polish National Multicenter Health Survey (WOBASZ). Subjects` daily food consumption was estimated by the 24-hour recall method and was continued for 3 years. It was determined on this basis, that 96 plant foods and plant food products were consumed. The own dietary database of polyphenol contents in food was used to calculate polyphenol intakes in the subjects. The daily total polyphenol intakes were calculated for both genders in individual age categories: 20-40 years, 41-60 years and 61-74 years. RESULTS: The average polyphenol consumption for the men was 1172 mg/day, and for the women it made 1031 mg/day. Plant food categories such as beverages, vegetables, fruits and cereals were found to be significant sources of polyphenols, of which tea, coffee, potatoes, apples and white bread were the main contributors. CONCLUSIONS: The amount of polyphenol consumption in Polish adult population is similar to the polyphenol intake in other countries populations, accounting roughly for 1g of polyphenols for both genders and different age groups. Patterns of polyphenol consumption, however, vary for genders and age groups. Polyphenols are characterized by a spectrum of antioxidant capabilities, therefore future studies should focus on dietary intakes of individual polyphenols.

Kinetics of fumonisin B1 formation in maize ears inoculated with Fusarium verticillioides.

The kinetics of fumonisin B1 (FB1) biosynthesis have been examined in ears of four botanical varieties Zea mays var. indentata, Zea mays var. indurata, Zea mays var. saccharata and Zea mays var. everta inoculated with F. verticillioides isolates at silking stage. The level of mycotoxin accumulated in kernels was correlated with sample harvest time (r = 0.73) and ergosterol content (r = 0.70). FB1 biosynthesis was influenced by amylose, starch and moisture contents, which undergo dynamic changes during grain formation. A comparative analysis of ear infection rates and efficiency of FB1 biosynthesis showed that these are separate characters and their expression could be independent.

Formation of fumonisins and other secondary metabolites by Fusarium oxysporum and F. proliferatum: a comparative study.

The principal aim of this study was to estimate the formation of fumonisins (FB(1) and FB(2)), moniliformin (MON), and ergosterol (ERG) by Fusarium oxysporum and Fusarium proliferatum, while the formation of beauvericin (BEA) was estimated by the latter Fusarium species only. Moreover, the effect of temperature on the biosynthesis of mycotoxins was also evaluated. Fumonisins were formed by F. proliferatum, with the highest yield at 18 degrees C (720.0-1976.6 microg g(-1) for FB(1), 74.2-670.8 microg g(-1) for FB(2)) and only by three of four F. oxysporum strains at a very low level (0.02-4.77 microg g(-1) for FB(1), 0.02-2.15 microg g(-1) for FB(2)). The amount of MON formed by F. proliferatum was the highest (p < 0.001) at 32 degrees C (3056.87 microg g(-1)), while MON biosynthesis by F. oxysporum was lower 227.54 microg g(-1) (p < 0.001). BEA was produced by F. proliferatum with the highest level at 25 degrees C (p < 0.001). ERG-recognized as an indicator of fungal biomass development and as a consequence of mycotoxin formation-was found at the highest concentration at a biosynthesis temperature of 25 degrees C for F. proliferatum and F. oxysporum (p < 0.001).

Reaction of winter wheat (Triticum aestivum L.) cultivars to infection with Fusarium spp.: mycotoxin contamination in grain and chaff.

This study compares the susceptibility of winter wheat (Triticum aestivum L.) cultivars to Fusarium head blight (FHB) and accumulation of mycotoxins in kernels and chaff under different climatic conditions in two locations-Cerekwica near Poznan (Central West Poland) and Sitaniec, near Zamosc, Lublin region (South East Poland). Very high variations were found in the concentrations of mycotoxins (zearalenone, ZEA; nivalenol, NIV; deoxynivalenol, DON; moniliformin, MON) in examined fractions: Fusarium-damaged kernels (FDK) and healthy looking kernels (HLK) and in chaff for individual cultivars in both locations. In most cases, significantly higher concentrations of investigated toxins were recorded in wheat from the area of Lublin than from Poznan (p < 0.05). The highest Fusarium infection rates and mycotoxin biosynthesis levels were observed in the Lublin location, with the percentage of the FDK fraction ranging 8.1-81.6. In this region, ZEA concentration (microg g(-1)) after inoculation with F. culmorum and F. graminearum ranged from 0.02-0.48 and 0.32-1.04, respectively. In the Poznan area, the toxin concentrations were considerably lower, ranging 0.01-0.10 and 0.03-0.13 microg g(-1) for both isolates, respectively. The concentration of DON was significantly higher than ZEA or NIV levels. The levels of MON accumulation (microg g(-1)) in the FDK fraction were between 0.14 and 1.73 (Poznan area) and ND (not detected) to 2.51 (Lublin area). F. avenaceum infection rate ranged 7-35% in samples where the toxin was detected.

Mycotoxins and mycotoxicoses under climatic conditions of Poland.

Ochratoxin A (OA), zearalenone (ZON), moniliformin (MON) as well as trichotecenes and fumonisines (FUM) are naturally occurring contaminants of cereals and animal feed. They pose a health risk not only to humans but also to livestock and, as a consequence, may cause economical losses either due to unfavorable effects on domestic animals themselves or to an increased potential for health effects in human beings consuming mycotoxin-contaminated edible animal products. At present, large-scale studies are carried out in EU countries to determine a safe, admissible concentration of these toxins in cereals and their processed products. The aim of this review is to collect and summarize information concerning the properties, occurrence and toxicity of these mycotoxins.

Zebrafish Meis functions to stabilize Pbx proteins and regulate hindbrain patterning.

Homeodomain-containing Hox proteins regulate segmental identity in Drosophila in concert with two partners known as Extradenticle (Exd) and Homothorax (Hth). These partners are themselves DNA-binding, homeodomain proteins, and probably function by revealing the intrinsic specificity of Hox proteins. Vertebrate orthologs of Exd and Hth, known as Pbx and Meis (named for a myeloid ecotropic leukemia virus integration site), respectively, are encoded by multigene families and are present in multimeric complexes together with vertebrate Hox proteins. Previous results have demonstrated that the zygotically encoded Pbx4/Lazarus (Lzr) protein is required for segmentation of the zebrafish hindbrain and proper expression and function of Hox genes. We demonstrate that Meis functions in the same pathway as Pbx in zebrafish hindbrain development, as expression of a dominant-negative mutant Meis results in phenotypes that are remarkably similar to that of lzr mutants. Surprisingly, expression of Meis protein partially rescues the lzr(-) phenotype. Lzr protein levels are increased in embryos overexpressing Meis and are reduced for lzr mutants that cannot bind to Meis. This implies a mechanism whereby Meis rescues lzr mutants by stabilizing maternally encoded Lzr. Our results define two functions of Meis during zebrafish hindbrain segmentation: that of a DNA-binding partner of Pbx proteins, and that of a post-transcriptional regulator of Pbx protein levels.

[Changes in dietary habits and selected cardiovascular risk factors of the Warsaw population over the 5-year observation period (1993-97/98)]. / Zmiany sposobu zywienia i wybranych czynników ryzyka chorób ukladu krazenia mieszkanców prawobrzeznej Warszawy w okresie 5-letniej obserwacji (1993-97/98).

The aim of the study was to characterise changes in dietary habits and cardiovascular risk factors of the Warsaw cohort population over a 5-year observation period. Two Warsaw population samples (249 men and 259 women), aged 35-64, were surveyed in 1993 and resurveyed in years 1997/98. Over a five-year observation period, daily dietary cholesterol decreased from 363 mg to 319 mg in men and from 263 mg to 232 mg in women, the percentage of energy from fat decreased from 38.8% to 34.7% in men and from 37.1% to 31.2% in women, the percentage of energy from saturated fatty acids from 13.6% to 11.7% and from 12.9% to 10.5% respectively. These changes resulted in a significant reduction in atherogenicity of the average diet as expressed by Keys score. Intakes of vitamins B1 and B2, calcium, magnesium and iron was consistently below Recommended Dietary Allowances (RDA) for the Polish population. Participants of both genders consumed significantly greater amounts of total bread (35%), milk (50%) and butcher meat products (22%). In the same time increase of HDL-cholesterol level (average of 2.0 mg/dl in men and 3.1 mg/dl in women) and decrease of LDL-cholesterol by 5 mg/dl in men was noted.

Ochratoxin A in porcine blood and in consumed feed samples.

The aim of the study was to estimate occurrence of ochratoxin A (OA) in feeds and the metabolite residues in porcine blood serum in Poland. Samples were collected in the period from February to May, 1999, in the southern Wielkopolska region. Altogether 40 and 45 samples of feed and porcine blood serum, respectively, were analyzed for OA. Percentage of samples contaminated with OA, both in case of feeds and blood, collected in the winter season was considerably higher than that for the spring season. The percentages for feeds were as follows: 47.6 and 26.3 %, while for porcine serum: 66.7 and 50.0 %, respectively winter and spring. In 25 % of cases ochratoxin A was present in both types of investigated material (feed, blood), whereas in 27.5 % of samples this metabolite was detected in blood only, or in 7.5 % only in the feed. The presence of OA was found neither in the feed nor in the serum in 40 % of all cases. In subgroups (feed, blood) the concentration in the whole collective of positive samples were in the range 0.3-13.5 ng/g and 0.3-69.5 ng/ml, respectively, while median values were 2.3 ng/g and 6.0 ng/ml. Only one feed and three porcine serum samples, were found to be contaminated at concentration levels higher than 10 ng/g or 10 ng/ml.

Phosphorylation of the cap-binding protein eukaryotic translation initiation factor 4E by protein kinase Mnk1 in vivo.

Eukaryotic translation initiation factor 4E (eIF4E) binds to the mRNA 5' cap and brings the mRNA into a complex with other protein synthesis initiation factors and ribosomes. The activity of mammalian eIF4E is important for the translation of capped mRNAs and is thought to be regulated by two mechanisms. First, eIF4E is sequestered by binding proteins, such as 4EBP1, in quiescent cells. Mitogens induce the release of eIF4E by stimulating the phosphorylation of 4EBP1. Second, mitogens and stresses induce the phosphorylation of eIF4E at Ser 209, increasing the affinity of eIF4E for capped mRNA and for an associated scaffolding protein, eIF4G. We previously showed that a mitogen- and stress-activated kinase, Mnk1, phosphorylates eIF4E in vitro at the physiological site. Here we show that Mnk1 regulates eIF4E phosphorylation in vivo. Mnk1 binds directly to eIF4G and copurifies with eIF4G and eIF4E. We identified activating phosphorylation sites in Mnk1 and developed dominant-negative and activated mutants. Expression of dominant-negative Mnk1 reduces mitogen-induced eIF4E phosphorylation, while expression of activated Mnk1 increases basal eIF4E phosphorylation. Activated mutant Mnk1 also induces extensive phosphorylation of eIF4E in cells overexpressing 4EBP1. This suggests that phosphorylation of eIF4E is catalyzed by Mnk1 or a very similar kinase in cells and is independent of other mitogenic signals that release eIF4E from 4EBP1.

The phosphorylation of eukaryotic initiation factor eIF4E in response to phorbol esters, cell stresses, and cytokines is mediated by distinct MAP kinase pathways.

Initiation factor eIF4E binds to the 5'-cap of eukaryotic mRNAs and plays a key role in the mechanism and regulation of translation. It may be regulated through its own phosphorylation and through inhibitory binding proteins (4E-BPs), which modulate its availability for initiation complex assembly. eIF4E phosphorylation is enhanced by phorbol esters. We show, using specific inhibitors, that this involves both the p38 mitogen-activated protein (MAP) kinase and Erk signaling pathways. Cell stresses such as arsenite and anisomycin and the cytokines tumor necrosis factor-alpha and interleukin-1beta also cause increased phosphorylation of eIF4E, which is abolished by the specific p38 MAP kinase inhibitor, SB203580. These changes in eIF4E phosphorylation parallel the activity of the eIF4E kinase, Mnk1. However other stresses such as heat shock, sorbitol, and H2O2, which also stimulate p38 MAP kinase and increase Mnk1 activity, do not increase phosphorylation of eIF4E. The latter stresses increase the binding of eIF4E to 4E-BP1, and we show that this blocks the phosphorylation of eIF4E by Mnk1 in vitro, which may explain the absence of an increase in eIF4E phosphorylation under these conditions.

Mitogen-activated protein kinases activate the serine/threonine kinases Mnk1 and Mnk2.

Mitogen-activated protein (MAP) kinases bind tightly to many of their physiologically relevant substrates. We have identified a new subfamily of murine serine/threonine kinases, whose members, MAP kinase-interacting kinase 1 (Mnk1) and Mnk2, bind tightly to the growth factor-regulated MAP kinases, Erk1 and Erk2. MNK1, but not Mnk2, also binds strongly to the stress-activated kinase, p38. MNK1 complexes more strongly with inactive than active Erk, implying that Mnk and Erk may dissociate after mitogen stimulation. Erk and p38 phosphorylate MNK1 and Mnk2, which stimulates their in vitro kinase activity toward a substrate, eukaryotic initiation factor-4E (eIF-4E). Initiation factor eIF-4E is a regulatory phosphoprotein whose phosphorylation is increased by insulin in an Erk-dependent manner. In vitro, MNK1 rapidly phosphorylates eIF-4E at the physiologically relevant site, Ser209. In cells, Mnk1 is post-translationally modified and enzymatically activated in response to treatment with either peptide growth factors, phorbol esters, anisomycin or UV. Mitogen- and stress-mediated MNK1 activation is blocked by inhibitors of MAP kinase kinase 1 (Mkk1) and p38, demonstrating that Mnk1 is downstream of multiple MAP kinases. MNK1 may define a convergence point between the growth factor-activated and one of the stress-activated protein kinase cascades and is a candidate to phosphorylate eIF-4E in cells.

Mitogen and stress response pathways: MAP kinase cascades and phosphatase regulation in mammals and yeast.

Evolutionarily conserved from yeast to man, mitogen-activated protein kinase (MAPK) pathways respond to a variety of disparate signals which induce differentiation, proliferation, or changes in intracellular enzyme regulation. Recent advances have identified two new mammalian MAPK relatives, JNK1 and p38, and the pathways which are responsible for their activation.

[Evaluation of dietary changes during a 10 year observation period of the local Pol-MONICA population in Warsaw with regard to risk of cardiovascular diseases]. / Ocena zmian sposobu zywienia w okresie 10-letniej obserwacji populacji miejskiej pol-MONICA Warszawa w aspekcie ryzyka chorób ukladu krazenia.

In the frame of the Warsaw Pol-MONICA Project three population screenings were performed in randomly selected 35-64 year men and women in 1984 (2571 participants), in 1988 (1397) and 1993 (1485). Twenty-four-hour dietary recalls were used for assessment of nutritional value of daily food ration and food products consumption. Statistical analyses was performed by linear regression method (SAS REG procedure). Substantial changes in food consumption pattern were noted during 10 years, particularly in the last 4 years. In 1993 participants of both sexes consumed significantly less cereals, butter, eggs (trend p < 0.01) and more fats of vegetable origin (trend p < 0.01). This changes reflected in nutritional values of daily food ration from 1984 to 1993: dietary cholesterol decreased from 630 mg to 433 mg in men and from 447 to 284 in women, percent of energy from saturated fatty acids decreased from 15.7% to 14.7% in men and from 15.9% to 13.9% in women, percent of energy from polyunsaturated fatty acids increased from 4.1% to 5.4% and from 4.3% to 5.5% respectively. Above mentioned changes resulted in a significant reduction of atherogenicity level of diet as expressed by Keys score. A period covered by the study was a period of economical changes in Poland associated with an introduction of a free food-market. Changes in food consumption and nutrient intake reflect the mixture of industry, mass communication patterns, and commercial advertising.

Evolutionary conservation of Xenopus laevis mitogen-activated protein kinase activation and function.

Saccharomyces cerevisiae possesses at least four mitogen-activated protein (MAP) kinase family members, encoded by the FUS3, KSS1, HOG1, and MPK1 genes, that participate in three distinct signaling pathways. We have tested whether a MAP kinase from Xenopus laevis (Xp42) can function in budding yeast, by expressing wild-type and mutant forms of Xp42 in different strains of S. cerevisiae. In Xenopus cells, Xp42 is phosphorylated on threonine188 and tyrosine190 when activated by a MAP kinase kinase (MAPKK). In S. cerevisiae, Xp42 is constitutively phosphorylated on tyrosine190. Since a kinase-inactive mutant of Xp42 is also phosphorylated, this phosphorylation is presumably due to activation by an endogenous MAPKK. Xp42 phosphorylation and kinase activity are dependent on yeast Bck1p, a putative MAPKK kinase (MAPKKK) and indirect upstream activator of Mpk1p. The loss of either Ste7p or Pbs2p, the upstream activators of Fus3p, Kss1p, and Hog1p, does not decrease the phosphorylation stoichiometry of Xp42. We also show that expression of Xenopus MAP kinase permits an mpk1::TRP1 deletion strain to grow at 37 degrees C. We conclude that S. cerevisiae and X. laevis possess evolutionarily conserved cascades, where biochemical activation and substrate specificity of MAP kinase have been maintained.

[Daily fat consumption by the population of Warsaw]. / Spozycie tluszczu w codziennej diecie ludnosci Warszawy.

Fat intake with the diet has been assessed within screening Pol-MONICA--Warsaw Project in randomly selected sample of Warsaw inhabitants aged between 35 and 64 years. Altogether 2571 individuals have been screened. Fat intake in daily diet has been estimated by the 24-hour recall technique. In men, fat intake covered 38.4% of the total energy supply with the diet whereas in female population - 37.7%. Mean contribution of SFA and P/S ratio in both sex groups have been 15.8% and 0.285, respectively. Daily cholesterol intake has been 641 mg (224 mg/1000 kcal) in male and 456 mg (227/1000 kcal) in female population. These values did not change significantly with the age of the screened individuals. Such a diet may be considered atherogenic.

Dietary habits of the middle-aged Warsaw population in 1984 relative to nutritional guidelines.

The dietary habits of 1265 men and 1309 women aged 35-64 years were assessed by food frequencies and 24-h recall. The consumption of fat was high relative to Recommended Dietary Allowances (mean 137% of RDA in the diet of men and 108% in women). The diet was deficient in carbohydrates (80 and 67% respectively), calcium (81 and 64%), vitamin A (84% for both sexes), vitamin B1 (91 and 74%), and vitamin C (41 and 30%). Energy, protein, dietary fibre, iron and vitamin B2 were at the RDA or exceeded it in the men's diet but were insufficient in the women's diet. Fat accounted for 38.5% (men) and 37.7% (women) of energy, with 15.8% of the energy derived from saturated fatty acids and 4.2% from polyunsaturates. The amount of cholesterol consumed was 641 and 452 mg, respectively. This type of diet may contribute to increased incidence of ischaemic heart disease. Only three meals a day were taken by over 83% of the respondents, with over 5-h intervals between meals which may, in addition, contribute to obesity.