Different kinds of cereal flours submitted to various technological treatments were classified on the basis of their mid-infrared spectra by pattern recognition techniques. Classification in the wavelet domain was achieved by using the wavelet packet transform for efficient pattern recognition (WPTER) algorithm, which allowed singling out the most discriminant spectral regions. Principal component analysis (PCA) on the selected features showed an effective clustering of the analyzed flours. Satisfactory classification models were obtained both on training and test samples. Furthermore, mixtures of varying composition of the studied flours were distributed in the PCA space according to their composition.

Among thiamin-binding proteins that ubiquitously occur in plant seeds, that of common buckwheat became a model of extensive studies of the chemical mechanism of ligand-protein interaction. In this work, the polypeptide components of buckwheat seed thiamin-binding protein (BSTBP) are identified and characterized. We suggest that BSTBP is probably a fraction of major storage 13 S globulin (legumin), has an average molecular mass of 235 kDa and comprises hexamers of 57-kDa and 38-kDa subunits in variable combinations. Each subunit is a pair of disulfide-linked polypeptide chains, 36 kDa plus 24 kDa and two-times 22 kDa, respectively. The N-terminal sequences of 22-kDa and 24-kDa components show strict homology with those reported for "basic subunits" of buckwheat legumin. By photoaffinity labeling of BSTBP with 4-azido-2-nitrobenzoylthiamine, it is shown that the 36-kDa chain plays the major role in thiamin binding, but the other chains may also be variably involved. Putative thiamin-binding fragments are identified and sequenced.

Various one- and two-step extraction procedures with and without a short application of ultrasound at the beginning of the extraction were used to examine the effect of sonication on the extractibility of the hemicellulose components of buckwheat hulls. The polysaccharides recovered from the extracts were characterised by yield as well as composition determined by chemical methods and spectroscopic techniques. They comprised a complex of glucuronoxylan and co-extracted amylose-rich starch in various proportions contaminated with other cell wall components (protein, pectic polysaccharides). The hemicellulose fractions obtained by classical and ultrasound-assisted extraction exhibited significant immunomodulatory activities. The increased yield of ultrasonically extracted hemicelluloses, which have preserved their structural and molecular properties as well as immunological activity, confirmed the importance and great potential of ultrasound-assisted extraction of industrially important polysaccharides from different tissues of plant materials.

We evaluated the nutritional factors of underutilized cereals (spelt, emmer, einkorn, millet, foxtail millet, semiperennial rye, naked oat, and naked barley) and buckwheat. The basic food components as well as minor nutrients were determined. The analyses included dry matter, ash, protein, dietary fiber, fat, fatty acids, amino acids, minerals, and lipophilic and hydrophilic vitamins. Rutin was also determined in buckwheat. We hope to offer new recipes for the healthy food production and for special dietary use (diabetes, celiac disease, phenylketonuria diet, etc.). Use of the germinated seeds is also suggested. The examples of some healthy food products in the Czech Republic are mentioned.

Female rats were examined for the effects of feeding buckwheat protein extract (BWPE) on the development of mammary tumor caused by administration of 7,12-dimethylbenz[alpha]anthracene. The percentage of rats with palpable mammary tumors and serum estradiol were lower in the BWPE-fed animals than the casein-fed ones, implying that BWPE intake retarded the mammary carcinogenesis by lowering serum estradiol.

Sucrose density gradient centrifugation showed that approximately 30% of total buckwheat proteins migrated with a 2S sedimentation coefficient. The main part of that fraction, polypeptides in the range of molecular mass from 8 to 16 kDa, were water soluble and represented albumins. SDS-PAGE analysis in nonreducing and reducing conditions showed that these polypeptides were not linked by disulfide bonds. The albumins make 25% of total salt soluble proteins, but that content is dramatically reduced under S-deficiency conditions. Determination of amino acid composition showed high methionine (9.2%) and lysine (5.6%) contents. That characteristic offers the possibility of transfer of the genes for individual albumin polypeptides to legumes and cereals limited in those essential amino acids to improve their nutritional quality.

The content of the flavonoid rutin was determined in different milling fractions of buckwheat seeds and in buckwheat stems, leaves, and flowers. The extraction was performed by using a solvent containing 60% of ethanol and 5% of ammonia in water. The extracts were analyzed by capillary electrophoresis (running buffer of 50 mM borate (pH 9.3), 100 mM sodium dodecyl sulfate; determination at 380 nm). In bran fractions the concentration of rutin was 131-476 ppm, and in flour fractions 19-168 ppm. On average, about 300, 1000, and 46000 ppm of rutin were found in leaves, stems, and flowers, respectively. The results indicate that buckwheat could be an important nutritional source of flavonoids, especially in countries with a low mean daily flavonoid intake.

Six flavonoids have been isolated and identified in buckwheat grain. These are rutin, orientin, vitexin, quercetin, isovitexin, and isoorientin. Rutin and isovitexin are the only flavonoid components of buckwheat seeds while hulls contain all six identified compounds. The total flavonoid concentration in the seeds was 18.8 and in the hulls 74 mg/100 g of dry matter. Dehulling the grain by using different temperature regimes resulted in drastic reductions of the total flavonoid concentration in the grain (by 75% of the control) and smaller but significant (15-20%) reduction in the hulls.

Growing rats were examined for the influence of a buckwheat protein diet on muscle weight and protein. In experiment 1, the rats were fed on a diet containing either casein or a buckwheat protein extract (BWPE) as the protein source (10%, 20% or 30%) for 5 wk. The relative weights (g per kg of body wt) of the gastrocnemius, plantaris and soleus muscles were higher in the BWPE-fed animals than in the casein-fed ones, but were unaffected by the dietary level of protein. These differences were not associated with growth. In experiment 2, the rats were fed on either a casein or BWPE diet at the 20% protein level for 5 wk. BWPE intake significantly elevated the gastrocnemius muscle weight, carcass protein and water, and reduced carcass fat. These results demonstrate that BWPE consumption causes muscle hypertrophy, elevates carcass protein and water, and reduces body fat.

The effects of bio-active peptide (BP) from buckwheat pollen and its synthetic analogues on murine and human lymphocytes and HL-60 cells were examined. Both natural and synthetic BP had the same effect in the lymphocyte blast transformation test. The proliferative response of HL-60 cells, co-cultured for 72 h with BP, was measured by the MTT method. BP had a stimulatory effect on HL-60 cell proliferation. The optical density (OD) value was 0.339+/-0.015, whereas, that of the control without BP was significantly lower (0.279+/-0.037; P < 0.05). The CD25 (IL-2Ralpha chain) positive cells of BP-stimulated human lymphocytes assessed by indirect immunofluorescence were 53.97+/-7.12%(1 microg/ml) and 42.53+/-4.4%(10 microg/ml), significantly higher than the control (11.77+/-1.88%; P < 0.001). The content of sIL-2R in the supernatant after culture with BP was determined by ELISA and was 1782.3+/-78.18 U/ml, lower than control 2381.2+/-99.9 U/ml. Taken together, our results showed that BP has an immunostimulatory effect on lymphocytes. The likely mode of BP action and the possibility of developing a novel immunomodulator are discussed.

A thiamin-binding protein from buckwheat (Fagopyrum esculentum Moench) seeds gave two bands of 56-and 50-kDa in the absence of 2-mercaptoethanol and a single band of 25-kDa in the presence of 2-mercaptoethanol on sodium dodecylsulfate gel electrophoresis. These results indicate that the protein consists of polypeptides linked by disulfide bond(s). The protein isolated from buckwheat seeds did not have immunological homology with the thiamin-binding proteins from rice seeds and sesame seeds. However, the binding of the protein to thiamin was inhibited by the modification of the carboxyl residues in the protein as well as that of the thiamin-binding protein from rice seeds. These results suggest that the thiamin-binding protein from buckwheat seeds differ from those from rice seeds and sesame seeds as to subunit structure or immunological properties, but resembles them in the mechanism of binding thiamin.

A study on the physico-chemical properties and structure together with the evaluation of starch digestibility was carried out on starch isolated from buckwheat (Fagopyrum esculentum) cultivated in different Italian areas. Results showed that buckwheat samples analysed were different among them and from wheat starch used as reference. Buckwheat granules were polygonal in shape and had a smaller diameter than the wheat starch granule. The starch obtained from buckwheat had a higher swelling power than the wheat one, probably as a consequence of the wheaker but more extensive bonding forces in the granule. During cooling, buckwheat samples showed a good paste stability.