Antitumoural activity of viniferin-enriched extracts from Vitis vinifera L. cell cultures.

The aim of this work was to evaluate the effect of stilbenes from different cultivars of Vitis vinifera on tumour proliferation. Extracts were obtained from elicited V. vinifera cell cultures and characterised by HPLC/DAD/MS. Cell growth was evaluated in four human cancer cell lines and in normal human fibroblasts. The cells were exposed to the extracts or to trans-resveratrol, used as reference molecule, for 48 h, at 1-10 µM concentrations of total stilbenoids. All the extracts exhibited antiproliferative activity, mediated by modulation of the cell cycle and induction of cytotoxicity in cancer but not in normal cell lines, and positively correlated with the content in dimeric stilbenoids. The Alphonse Lavallée extract was the most active, and the obtained stilbenoid fraction resulted 8-10 times more active than trans-resveratrol. Extracts from V. vinifera cell cultures could represent new sources of active stilbenoid compounds to be further assayed in in vivo studies for their antitumoural properties.

Stilbene production in cell cultures of Vitis vinifera L. cvs Red Globe and Michele Palieri elicited by methyl jasmonate.

Cell cultures obtained from Vitis vinifera cvs Michele Palieri and Red Globe were cultured in order to stimulate stilbene production. In the calli, stilbene production peaked at day 22 of culture for both cultivars; the main compound was trans-piceid, followed by cis-piceid. Methyl jasmonate, which was added to cell suspensions in the first half of the exponential growth phase, enhanced stilbene accumulation, producing mainly trans-piceid and epsilon-viniferin. Other stilbenoids, though in lower quantities, were identified by liquid chromatography/positive electrospray mass spectrometry. epsilon-Viniferin and trans-resveratrol were the main compounds released into the culture medium. The total quantity of stilbenes was genotype dependent, with a better response found for the cv Red Globe.

Chitosan enhances xanthone production in Hypericum perforatum subsp. angustifolium cell cultures.

Hypericum perforatum is an important medicinal plant containing numerous biologically active compounds. The effect of chitosan elicitation on xanthone biosynthesis in calli and in cell suspension cultures of H. perforatum subsp. angustifolium was evaluated. Elicited cell cultures showed an increase in xanthone production and a simultaneous decrease in flavonoid production. Chitosan also induced the production of 1,7-dihydroxyxanthone (euxanthone) and cadensin G, which were not detected in either the calli nor the non-elicited cell cultures. 1,7-Dihydroxyxanthone was in part (21%) released in the culture medium.

Anthocyanins and flavan-3-ols from grapes and wines of Vitis vinifera cv. Cesanese d'Affile.

The objective of the present study was to evaluate the amount of some potential health-promoting phenols in the grape of Vitis vinifera cv. Cesanese d'Affile and in wines made from these grapes. The analyses were performed using HPLC/DAD/MS. The accumulation of anthocyanins in the skin and flavan-3-ols in the seed was determined at different stages of ripening of the grape (i.e. green, veraison, middle stage of ripening, and complete ripening). Thirteen anthocyanins were identified in the skin at all stages of ripening, except the green stage. With regard to flavan-3-ols, (+)-catechin, (-)-epicatechin, and (-)-epicatechin gallate were detected in all of the seed samples. The highest (+)-catechin content was found in the seeds of the green grape (2 mg g(-1) DW), whereas in the seeds from the completely ripe grape the content was more than ten times lower. The highest catechin content in the seed was correlated with the lowest anthocyanin content in the skin. The wines produced in the years 2004 and 2005 showed, at wavelengths of 520 and 280 nm, almost identical quali-quantitative chromatographic profiles, with high concentrations of anthocyanin 3-O-glucosides, low concentrations of acylated anthocyanins, and trace amounts of (+)-catechin and (-)-epicatechin.

CPT accumulation in the fruit and during early phases of plant development in Camptotheca acuminata Decaisne (Nyssaceae).

We describe the dynamics of camptothecin (CPT) accumulation during the early phases of plant development in Camptotheca acuminata. TLC and HPLC analyses were performed on the entire mature fruit and all of its parts and on the organs of seedlings in different developmental stages. In the mature fruit, the CPT content was relatively high (2.83 mg g(-1) DW); it was accumulated especially in the endosperm (1.82 mg g(-1) DW) and embryo (1.10 mg g(-1) DW). Regarding seedlings, CPT was present in all of the organs, at all developmental stages. In the cotyledons, content was greatest in the early developmental stages (4.23 mg g(-1) DW) and drastically decreased once the organ reached maturity (0.34 mg g(-1) DW). A similar trend was observed for the true leaves. The results suggest that CPT is translocated from the organs at an advanced developmental stage to those at an early stage.

Triterpenoids and ellagic acid derivatives from in vitro cultures of Camptotheca acuminata Decaisne.

The metabolic profile of secondary products in calli and cell suspension cultures of Camptotheca acuminata Decaisne was investigated and compared to that of the leaves and roots taken from the plant. Neither in vitro system produced the anticancer quinoline alkaloid camptothecin (CPT); they accumulated discrete quantities of polyhydroxylated triterpenoids, different from those found in the plant organs, and ellagic acid derivatives. Nine ellagic acid derivatives (1a-1d and 2a-2e) and eight triterpenoid acids (3a-3e and 4a-4c) were isolated and characterised. All the identified triterpenes were related to ursane- or oleanane-type skeletons and their concentrations rose to 4.5% in suspended cells.

Serum cystatin C as a reliable marker of changes in glomerular filtration rate in children with urinary tract malformations.

PURPOSE: Cystatin C has been suggested as a simple method of estimating GFR more accurately than creatinine in children. We compared the diagnostic accuracy of cystatin C with serum creatinine and the Schwartz formula for estimating GFR in patients with UTMs. MATERIALS AND METHODS: We prospectively compared 72 patients with UTMs (20 days to 36 months old, 58 males and 14 females) with a group of 72 healthy controls (10 days to 48 months old, 53 males and 19 females). All patients underwent nuclear medicine clearance investigations with (99m)Tc DTPA. RESULTS: Serum concentration of cystatin C revealed a higher correlation with (99m)Tc DTPA (r = 0.62, p <0.001) than serum concentration of creatinine (r = 0.30, p <0.01) or Schwartz formula (r = 0.51, p <0.001). These results were more evident in patients with uropathy (19) with mild renal impairment. Agreement between methods was assessed using Bland Altman analysis. Mean differences between GFR calculated with (99m)Tc DTPA and cystatin C based GFR estimation or Schwartz formula were -2.6% +/- 46.7% and -73.4% +/- 53.6%, respectively. Diagnostic accuracy in identifying decreased GFR measured as AUC was always highest for cystatin C but hardly sufficient for the 3 variables. Cystatin C performed better in the 0 to 6-month-olds (0.70 +/- 0.08 for cystatin C, 0.58 +/- 0.07 for Schwartz estimate) and patients older than 12 months (0.82 +/- 0.09 for cystatin C, 0.65 +/- 0.11 for Schwartz estimate). CONCLUSIONS: Cystatin C proved to be a superior marker rate over serum creatinine in estimating glomerular filtration in children younger than 3 years with UTMs and mild renal impairment, thus, offering a more specific and practical measure for monitoring GFR.

Accumulation of essential oils in relation to root differentiation in Angelica archangelica L.

The accumulation of essential oils in Angelica archangelica subsp. archangelica roots at different developmental stages was investigated through histochemical and chemical analyses. Roots less than 1 mm in diameter showed a primary diarch structure and two primary secretory ducts in the pericycle. These ducts were ephemeral and probably became dysfunctional early on. Oil accumulation was observed only in the secondary secretory ducts formed by cambium activity and located in the secondary phloem. Gas chromatographic analyses revealed that only taproots exceeding 5 mm in diameter contained a high concentration of alpha- and beta-phellandrene, which appreciably influence the oil's aroma.

Histological study of callus formation and optimization of cell growth in Taxus baccata.

Callus growth of Taxus baccata was optimized when Murashige and Skoog or B5 media were used with 2,4-D/kinetin ratios of 1:0.1, 2:0.1, and 5:0.1, when statistically equivalent results were obtained. A cell suspension culture was successfully achieved with a 3-fold increase in fresh weight after 40 days. Histological examination showed that in the leaf explants both the epidermis and mesophyll tissues divided to produce callus. In the stem explants callus formation was due to the cell division of the cortical parenchyma and of the cambium.

Comparison between metabolite productions in cell culture and in whole plant of Maclura pomifera.

Plant tissue cultures of Maclura pomifera showed a metabolite accumulation pattern which was both quantitatively and qualitatively different from that of the parent plant. Triterpenes and flavonoids were isolated from callus and cell cultures, however, xanthones and stilbenes, which have been reported in the whole plant, were not found. Among the flavonoids, flavones and flavanones were produced preferentially by the suspended cells, but with the prenyl substituents exclusively on ring A, while the isoflavones did not show the 3',4'-dihydroxyl substitution pattern found in the products isolated from fruits. A new prenylated 6'-deoxychalcone was also isolated from the callus and cell cultures.

Metabolites from in vitro cultures of Cassia didymobotrya.

From suspension cultures of Cassia didymobotrya 7-acetylchrysophanol, chrysophanol-physcion-l0,l0'-bianthrone, (E)- and (Z)-3'-hydroxy-3,4,5'-trimethoxystilbene, (E)-4,3'-dihydroxy-3,5'-dimethoxystilbene and 7,4'-dihydroxy-3,5,3'-trimethoxyflavone have been isolated along with several known metabolites.