Krzysztof Gulewicz

Food safety evaluation of broccoli and radish sprouts.

Three cultivars of broccoli seeds (Brassica oleracea var. italica), cv. Tiburon, cv. Belstar and cv. Lucky, and two cultivars of radish seeds (Raphanus sativus), cv. Rebel and cv. Bolide, were germinated for three and five days and safety aspects such as microbiological counts and biogenic amines were investigated. Cytotoxicity evaluation was also carried out. Broccoli and radish sprouts contained numbers of mesophilic, psychrotrophic, total and faecal coliform bacteria which are the usual counts for minimally processed germinated seeds. Putrescine, cadaverine, histamine, tyramine, spermidine and spermine increased during sprout production although these levels were below those permitted by legislation (5 mg/100 g of edible food). Broccoli and radish sprouts demonstrated no toxic effects on proliferation and viability of HL-60 cells and should be included in our diets as healthy and safe fresh foods.

Assessment of the effect of alpha-galactosides injected during embryogenesis on selected chicken traits

The effect of different doses of alpha-galactoside (RFOs) preparations from Pisum sativum L. cv. Opal, injected into eggs during embryogenesis, on maintaining a high number of bifidobacteria, selected chicken broiler traits and the lipoprotein level of blood were studied. Two independent experiments were conducted. In the first, Ringer water solution containing 1.763 mg/egg of fructooligosaccharides (FOS) (I group), 2.1158 mg of pea RFO preparation containing 20% sucrose (II group) and 0.4232 mg of sucrose (III group) were injected into Hubbard broiler breeder eggs containing 12-day old embryos. Only Ringer water solution was applied to the eggs of the control group (IV group). The number of bifidobacteria determined in faeces of two-day old chicken of groups I and II was significantly higher in comparison with the sucrose and control groups. The high level ofbifidobacteria of groups I and II was maintained during 6 weeks. The dose of both preparations had no influence on the body weight, carcass, breast muscle, leg and abdominal fat ratio, total cholesterol, HDL and LDL serum concentrations. Broiler mortality and breast muscle cholesterol concentration was highest (P < 0.05) for the control group. On the other hand, the European Production Index, as well as serum triglycerides, were the lowest for this group. The second experiment was performed on Hybro G chicken breeder eggs. 0.69, 3.43 and 6.87 mg/egg of pea RFO preparation doses containing 20% sucrose were injected into the experimental groups. The number of bifidobacteria in the caecum and selected meat traits of broilers were determined. The results of this experiment confirmed that RFO injection in ovo causes the long-time maintenance of a high level ofbifidobacteria. The dose of the preparations does not have any effect on the selected broiler meat traits, except that the highest dose increases the percent of carcase in body weight. However, this dose reduced the hatchability of the treated embryos.

Role of Nitrogen and Plant Growth Regulators in the Exudation and Accumulation of lsoflavonoids by Roots of Intact White (Lupin (Lupinus albus L.) Plants

Summary Two isoflavonoid glycosides (7-O-β-glucopyranosides of 2′-hydroxygenistein and genistein) and up to six aglycones (2′-hydroxygenistein, genistein, wighteone, luteone, lupinisoflavone A, parvisoflavone B) were identified in root exudates and/or root extracts from white lupin ( Lupinus albus L. cv. Bac) plants. Three-d-old seedlings were cultivated for 14d on 1/2 Knops nutrient solutions. Various treatments, differing in the form (No 3 − , NH 4 + , NH 4 N0 3 , urea) and concentration of nitrogen, and the presence of plant growth regulators in the medium were tested. Nitrogen, in all forms applied, affects both the exudation and accumulation of phenolics with the inhibitory activites arranged in the order: N0 3 − > NH 4 N0 3 > urea > NH 4 + . With higher concentration of N in the medium, the exudation of phenolics decreases. Processes of isoflavone exudation and accumulation respond differentially to nitrogen status. Exogenously applied plant growth regulators modify inhibitory effects of nitrate on the exudation and accumulation of isoflavones. Their activity depend on the type of growth regulator used (NAA, BAP, GA 3 ), its chemical structure (IAA, IBA, NAA) and concentration in the medium. Lupin isoflavones might be potential signals in legume — Rhizobium interactions. These data show a correlation between plant nutrient status and the accumulation and exudation of putative phenolic signal molecules, thus indicating the existence of possible additional controls in the regulation of nodulation process.

Anticancer activity of the Uncaria tomentosa (Willd.) DC. preparations with different oxindole alkaloid composition.

The activity of Uncaria tomentosa preparations on cancer cells was studied using in vitro and in vivo models. IC (50) values were calculated for preparations with different quantitative and qualitative oxindole alkaloid composition: B/W(37) --bark extracted in water at 37 °C, B/W(b)--bark extracted in boiling water, B/50E(37) --bark extracted in 50% ethanol at 37 °C, B/E(b)--bark extracted in boiling 96% ethanol, B/96E(37) --bark extracted in 96% ethanol at 37 °C and B/SRT--bark extracted in water and dichloromethane. Generally, the results obtained showed a high correlation between the total oxindole alkaloid content (from 0.43% to 50.40% d.m.) and the antiproliferative activity of the preparations (IC(50) from >1000 μg/ml to 23.57 μg/ml). B/96E(37) and B/SRT were the most cytotoxic preparations, whereas the lowest toxicity was observed for B/W(37). B/96E(37) were shown to be active against Lewis lung carcinoma (LL/2) [IC(50) =25.06 μg/ml], cervical carcinoma (KB) [IC(50) =35.69 μg/ml] and colon adenocarcinoma (SW707) [IC(50) =49.06 μg/ml]. B/SRT was especially effective in inhibiting proliferation of cervical carcinoma (KB) [IC(50) =23.57 μg/ml], breast carcinoma (MCF-7) [IC(50) =29.86 μg/ml] and lung carcinoma (A-549) [IC(50) =40.03 μg/ml]. Further animal studies on mice bearing Lewis lung carcinoma showed significant inhibition of tumor growth by B/W(37) administered for 21 days at daily doses of 5 and 0.5 mg (p=0.0009). There were no significant changes in the cell cycles of tumor cells with the exception of cell decrease at the G₂/M phase after the administration of B/96E(37) at a daily dose of 0.5 mg and the G(1)/G(0) cells cycle arrest demonstrated after the B/SRT therapy at a daily-dose of 0.05 mg. All tested preparations were non-toxic and well tolerated.

Lupins as a potential source of raffinose family oligosaccharides: Preparative method for their isolation and purification

Abstract Results of studies on the content of raffinose family oligosaccharides (RFOs) in plants of 11 species of the Leguminosae family are presented. It is clearly shown that plants of the Leguminosae family, especially of the lupin species, are the richest source of these sugars among the plants tested. A simple method of preparative isolation and purification of RFOs from bitter lupin extract was developed. This method seems to be useful for efficient isolation of oligosaccharides from plant material for further nutritional tests.

Utilization of the chicken embryo model for assessment of biological activity of different oligosaccharides

The effect of different oligosaccharides--alpha-galactoside preparations from Lupinus albus seeds differing in sucrose content, raffinose and fructooligosaccharides on the growth of chicken intestine microflora and the hatchability and weight of the treated embryos were studied. The assessment of biological activity of these oligosaccharides was done in ovo on the chicken embryo model. The eggs of experimental groups containing twelve days old embryos were injected into the air cell with 0.2 ml of Ringer water solution containing 0.1763; 0.8815 and 1.763 mg/egg of an oligosaccharide preparation, while the control group was injected with 0.2 ml of Ringer water solution only. All oligosaccharide preparations in higher doses had an influence on chicken hatchability and increased bifidobacteria in the colon of two day old chicken. The number of bifidobacteria depends significantly on the kind of oligosaccharide preparation used and its dose. For all experimental groups, the number of bifidobacteria was significantly higher in comparison to the control.

Protective activity of the Uncaria tomentosa extracts on human erythrocytes in oxidative stress induced by 2,4-dichlorophenol (2,4-DCP) and catechol

The purpose of this study was to evaluate the effect of the ethanolic and aqueous extracts of Uncaria tomentosa on human erythrocytes and additionally the assessment of protective effect of these extracts on hemolysis induction, hemoglobin oxidation, and changes in the level of reactive oxygen species (ROS) and lipid peroxidation, which were provoked by selected xenobiotics, i.e. 2,4-dichlorophenol (2,4-DCP) and catechol. All tested extracts, even at a very high concentration of 500 μg/ml were not toxic to the erythrocytes because they did not cause lipid peroxidation, increase methemoglobin and ROS levels nor provoked hemolysis. The results of this study also revealed protective effect of extracts of U. tomentosa. The extracts studied depleted the extent of hemoglobin oxidation and lipid peroxidation as well as decreased the level of ROS and hemolysis, which was provoked by 2,4-DCP. No protective activity of the extracts against catechol action, which is a precursor of semiquinones in cell was found. A difference in the effect of the extracts studied was observed. Ethanol-based extracts revealed more pronounced ability to inhibit oxidation processes in human erythrocytes.

Application of solid phase extraction for profiling quinolizidine alkaloids and phenolic compounds in Lupinus albus

Solid phase extraction on strong cation exchanger (benzene sulphonic groups) and reverse phase (octadecyl groups) cartridges connected in series was used for a one-step isolation of quinolizidine alkaloids and phenolic compounds present in extracts from lupin seedlings. Both classes of compounds were further analysed by gas chromatography-mass spectrometry. This technique was used to analyse changes in the alkaloid and isoflavonoid profiles during the defence reaction of lupin plants evoked by abiotic (CuCl2) and biotic (elicitor preparation from Colletotrichum lindemuthianumcell walls) factors.

The effect of lupin alkaloids and ethanol extracts from seeds of Lupinus angustifolius on selected bacterial strains

Summary The following pure alkaloids were isolated from lupin seeds ( L. angustifolius var. Mirela): lupanine, 13-OH-lupanine, angustifoline and a 48 % ethanol extract. They and the commercial sparteine were tested for bacteriostatic effects against pathogenic bacteria, namely, Gram-positive ( Staphylococcus aureus ATCC 25923, Bacillus subtilis ATCC 6633) and Gram-negative ( Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853); a strain of Gram-positive Bacillus thuringiensis HD-4 with sporeforming rods, isolated from soil, was also tested. The established bacteriostatic agents, e.g. antibiotics, are active at a concentration 3 to 4 orders of magnitude lower than the tested lupin alkaloids. Therefore a bacteriostatic effect is possible in plants under physiological conditions, but a pharmaceutical application of these products as bacteriostatic agents is rather unlikely. The presented data support the allelopathic function of alkaloids.

Changes in nutritional value and cytotoxicity of garden cress germinated with different selenium solutions.

The selenium supply in almost all European countries is below the recommended daily intake, and different strategies are followed to fortify foods. In the present work, the influence of germination of garden cress ( Lepidium sativum cv. Ogrodowa) in different selenium solutions (Na(2)SeO(3) and Na(2)SeO(4)) on Se uptake, total antioxidant capacity, glucosinolates, protein, and amino acids was studied. Cytotoxicity in HL-60 human leukemic cell line was also assessed. The addition of selenite (Na(2)SeO(3)) or selenate (Na(2)SeO(4)) led to a significant increment in Se uptake in garden cress sprouts, and the highest Se content was observed at 8 mg/L in both inorganic Se solutions (36-38 microg/g of dm). The Se-enriched sprouts presented a large total antioxidant capacity (142-157 mumol of Trolox/g of dm), total glucosinolate content (99-124 microg/g of dm), protein (36-37% dm), and total essential amino acid content (40-41 g/100 g of protein), and no cytotoxicity on HL-60 human leukemic cells was observed. Garden cress sprouts obtained with selenite solution at 8 mg/L presented the best nutritional qualities and might provide a substantial proportion of Se in European diets. Bearing in mind the high nutritional value of sprouts, these may serve for the production of functional foods.