Radosław Pilarski

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.

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.

Products and Biopreparations from Alkaloid-rich Lupin in Animal Nutrition and Ecological Agriculture

(Suppl.): 59-66.This paper presents a new approach to utilization of alkaloid-rich lupin as a high protein andecologicalplant.Afterprocessingofbitterlupinseeds,manyvaluableproductsareobtained,i.e. protein concentrate, high dietary fiber product, raffinose family oligosaccharides(RFOs), and lupin extract. The described debittering process is fully ecological withoutwaste and all obtained products may be utilized in different domains. Therefore, (i) the lupinprotein concentrate may be a soybean substitute for feeding animals. (ii) high dietary fiberproduct improves rheological properties of dough and quality of bakery products, (iii) RFOsare prebiotics, (iv) lupin extract may be used for ecological plant cultivation, as a plantprotection preparation and as a medium for the production of yeast, gibberellic acid (GA

Influence of Germination with Different Selenium Solutions on Nutritional Value and Cytotoxicity of Lupin Seeds

The effect of different selenium solutions during germination of lupin seeds (Lupinus angustifolius L. cv. Zapaton) on the content of total selenium, protein, amino acids, soluble carbohydrates, total antioxidant activity, and cytotoxicity on HL-60 human leukemic cell line has been studied. Seeds were germinated in the presence of selenite (Na2SeO3) or selenate (Na2SeO4) solutions at different concentrations (0, 2, 4, 6, and 8 mg/L) for 5 days at either 20 or 25 degrees C. The addition of inorganic Se forms significantly increased Se content in lupin sprouts in a dose-dependent manner. The highest Se content in lupin sprouts was observed when germination was carried out with selenate solutions at 20 degrees C (11 microg/g of dw) or 25 degrees C (14 microg/g of dw). The Se-enriched sprouts presented an improvement in antioxidant activity (up to 117.8 and 103.5 micromol of Trolox/g of dw) as well as in essential amino acid content, and no cytotoxicity was observed on HL-60 human leukemic cells. Lupin seeds germinated with 8 mg/L selenate solutions for 5 days at 20 degrees C exhibited a higher germination rate (>90%) and a higher concentration of some essential amino acids than those obtained in selenite solutions in the same germination conditions. Therefore, the employment of selenate solutions at a concentration of 8 mg/L and germination for 5 days at 20 degrees C may be suggested for the production of Se-enriched lupin sprouts.

Effect of Alkaloid-Free and Alkaloid-Rich preparations from Uncaria tomentosa bark on mitotic activity and chromosome morphology evaluated by Allium Test

ETHNOPHARMACOLOGICAL RELEVANCE Uncaria tomentosa (Willd.) DC. is the most popular Peruvian plant, used in folk medicine for different purposes. It contains thousands of active compounds with great content of alkaloids. AIM OF STUDY Two different fractions of Alkaloid-Rich and Alkaloid-Free were researched on chromosome morphology, mitotic activity and phases indexes. MATERIALS AND METHODS Cells of Allium Test (meristematic cells of root tips) were incubated up to 24h in different concentrations of Alkaloid-Free and Alkaloid-Rich fraction obtained from Uncaria tomentosa bark followed by 48 h of postincubation in water. The chromosome morphology was analyzed and the content of mitotic and phase indexes were done. Individual compounds, oxindole alkaloids, phenolic compounds and sugars were determined. RESULTS In Alkaloid-Rich and Alkaloid-Free fractions (different in chemical composition) we observed condensation and contraction of chromosomes (more in Alkaloid-Rich) with retardation and/or inhibition of mitoses and changed mitotic phases. Postincubation reversed results in the highest concentration which was lethal (in mostly Alkaloid-Rich fraction). CONCLUSIONS Our studies indicate that different action can depend on different groups of active compounds in a preparation either containing alkaloids or not. Other fraction analysis may be useful in the future.

Studies on the Influence of Different Nitrogen Forms on the Chemical Composition of Various Cultivars of Lupinus albus L.

The effect of nitrogen (N) forms on the chemical composition of three cultivars of Lupinus albus L. has been studied. The N form had a significant effect on accumulation of alkaloids, polyamines, and sugars dependent upon the cultivar. In general, the ammonium and nitrate (NH4 ++NO3 −) and amine (–NH2) forms in all cultivars caused visible increases in alkaloid content. In the case of cv. ‘Butan,’ the (NH4 + + NO3 −) and (NO3 −) forms caused significant increases in polyamine contents, whereas Bradyrhizobium lupini plus (NH4 + + NO3 −), N deficiency (Nd), and (NH4 +) forms caused a decrease of polyamine levels. The N forms did not influence polyamine contents as much in cvs. ‘Marta’ and ‘Multolupa.’ The Bradyrhizobium lupini plus (NH4 + + NO3 −), (NO3 −), (NH4 +), (NH4 + + NO3 −), and (–NH2) caused a decrease of total sugars in seeds of ‘Butan’ and ‘Marta’ cvs.