Dumitriţa Rugină

Anthocyanin determination in blueberry extracts from various cultivars and their antiproliferative and apoptotic properties in B16-F10 metastatic murine melanoma cells

Blueberry consumption is associated with health benefits contributing to a reduced risk for cardiovascular disease, diabetes and cancer. The aim of this study was to determine the anthocyanin profile of blueberry extracts and to evaluate their effects on B16-F10 metastatic melanoma murine cells. Seven blueberry cultivars cultivated in Romania were used. The blueberry extracts were purified over an Amberlite XAD-7 resin and a Sephadex LH-20 column, in order to obtain the anthocyanin rich fractions (ARF). The antioxidant activity of the ARF of all cultivars was evaluated by ABTS, CUPRAC and ORAC assays. High performance liquid chromatography followed by electrospray ionization mass spectrometry (HPLC-ESI-MS) was used to identify and quantify individual anthocyanins. The anthocyanin content of tested cultivars ranged from 101.88 to 195.01 mg malvidin-3-glucoside/100g fresh weight. The anthocyanin rich-fraction obtained from cultivar Torro (ARF-T) was shown to have the highest anthocyanin content and antioxidant activity, and inhibited B16-F10 melanoma murine cells proliferation at concentrations higher than 500 μg/ml. In addition, ARF-T stimulated apoptosis and increased total LDH activity in metastatic B16-F10 melanoma murine cells. These results indicate that the anthocyanins from blueberry cultivar could be used as a chemopreventive or adjuvant treatment for metastasis control.

Antiproliferative and Antioxidant Properties of Anthocyanin Rich Extracts from Blueberry and Blackcurrant Juice

The present study was aimed at evaluating the antiproliferative potential of anthocyanin-rich fractions (ARFs) obtained from two commercially available juices (blueberry and blackcurrant juices) on three tumor cell lines; B16F10 (murine melanoma), A2780 (ovarian cancer) and HeLa (cervical cancer). Individual anthocyanin determination, identification and quantification were done using HPLC-MS. Antioxidant activity of the juices was determined through different mechanism methods such as DPPH and ORAC. For biological testing, the juices were purified through C18 cartridges in order to obtain fractions rich in anthocyanins. The major anthocyanins identified were glycosylated cyanidin derivatives. The antiproliferative activity of the fractions was tested using the MTT assay. The antiproliferative potential of ARF was found to be associated with those bioactive molecules, anthocyanins due to their antioxidant potential. The results obtained indicated that both blueberry and blackcurrants are rich sources of antioxidants including anthocyanins and therefore these fruits are highly recommended for daily consumption to prevent numerous degenerative diseases.

Carotenoid and fatty acid profiles of bilberries and cultivated blueberries from Romania

The worldwide consumption of blueberries has increased due to their potential health effects. They contain micronutrients, various phytochemicals and anti-oxidant pigments, represented especially by anthocyanins. In addition to these, carotenoids and fatty acids are also present. The aim of this study was the identification and quantification of carotenoids (by HPLC) and fatty acids (by GC with flame ionisation detection) in two bilberry genotypes (Wild 1 and 2) and three cultivars (BlueCrop, Elliot, and Duke). The main carotenoids identified were lutein, β-cryptoxanthin, and β-carotene, with the average total carotenoid content of 266 μg per 100 g of fruit. The ratio of saturated fatty acids (SFAs) to polyunsaturated fatty acids (PUFAs) was 0.098 for Wild 1 and 0.11 for Wild 2 genotypes and ranged from 0.14 to 0.28 for cultivars. The main fatty acids identified were linoleic (18:2) and linolenic (18:3) acids in all the samples analysed. In Vaccinium myrtillus the amount of carotenoids and fatty acids was higher than in Vaccinium corymbosum species.

Pharmacologically Active Plant-Derived Natural Products

Abstract A mounting body of evidence strongly supports the theory that natural bioactive compounds (e.g., polyphenols, carotenoids, and stilbenes) present in vegetables and fruits are able to prevent several chronic diseases. However, a large number of bioactive compounds that are linked to health benefits have low stability, are poorly absorbed, and own very limited bioavailability in humans, hindering positive results in clinical applications. Accordingly, a desirable strategy is to enhance their bioavailability or to increase their solubility, stability, and release, by developing efficient delivery systems for oral or topical administration. In this chapter, we focus on some relevant progresses on the development of delivery carriers designed to improve bioavailability of bioactive compounds like polyphenols, carotenoids, and stilbenes, with potential medical applications.

Chromatographic analysis of lypophilic compounds in eggs from organically fed hens

SUMMARY Egg yolks represent a common foodstuff in the human diet and are an important source of nutrients including lipids and carotenoids. The aim of this study was to compare the carotenoid, fatty acid and vitamin E contents of the yolk of eggs from nine hen breeds (Barred Plymouth Rock, Speckled Italian, Black Italian, Red Italian, Rhode Island, Silver Laced Wyandotte, Gold Araucana, Partridge Brahma and Yellow Cochin) raised in barns in an enclosed house. Gas chromatography‐mass spectrometry (GC‐MS) was used for the fatty acid analyses, and reversed phase‐high performance liquid chromatography‐photodiode array detection (RP‐HPLC‐PDA) for carotenoids and vitamin E quantification. The major carotenoids identified were lutein and zeaxanthin, which together represented more than 93% of the total carotenoids, followed by &bgr;‐cryptoxanthin and &bgr;‐carotene. The results indicated that the total carotenoid content of the eggs ranged from 16.84‐87.31 &mgr;g/g egg yolk (average value 49 &mgr;g/g egg yolk). The most representative fatty acids found were palmitic acid, stearic acid, oleic acid and linoleic acid. Significant differences in the proportions of &agr;‐linolenic acid were observed among the samples. The &agr;‐tocopherol content was directly correlated with the total carotenoid content. The results show that the chemical composition of egg yolk varies greatly among hen breeds.

Antiproliferative and Apoptotic Potential of Cyanidin-Based Anthocyanins on Melanoma Cells

Elderberries are known for their high anthocyanins content, which have been shown to possess anti-proliferative and anti-cancer effects. Anthocyanins enriched extract (AEE) was obtained from elderberries and was characterized by LC/DAD/ESI-MS analysis. Five cyanidin-based anthocyanins were identified, among which Cy-3-O-samb was the major compound (51%). The total anthocyanins content of AEE was 495 mg Cy-3-O-samb/100 g FW. AEE inhibited proliferation of metastatic B16-F10 murine melanoma cells, in a concentration-dependent manner, with an IC50 of 264.3 μg/mL. LDH (lactate dehydrogenase), as a marker of membrane integrity, increased 74% in B16-F10 cells treated with 250 μg/mL AEE, compared to control. It was observed that apoptosis is the mechanism of melanoma cell death after AEE treatment, confirmed morphologically by acridine orange/ethidium bromide double staining and TUNEL analysis. These results indicate that elderberry-derived anthocyanins might be utilized in future applications as topical adjuvant in skin cancer therapy.

Resveratrol Modulates Oxidative Status in Rose Bengal Photosensitized Retinal Pigment Epithelial Cells

Eye exposure to high light intensities can produce a photochemical damage to retinal pigment epithelium (RPE), leading to severe pathologies. RPE D407 cells treated with Resveratrol (RSV, 100 μM, 24h) +/photosensitizer Rose Bengal (RB, 500 nM, 1h) were exposed to green LED light. Cell viability and level of intracellular reactive oxygen species (ROS) produced after exposure to green light +/RB were evaluated by MTT, respectively ROS assays. Spectrophotometric methods were used to determine GSH level and enzyme activities (CAT, SOD, GPx). Green LED light alone reduced cell viability with 30%, but a 40% reduction was observed in the presence of RB. RSV proved to have a strong positive impact, with a 20% increase in the viability of D407 cells exposed to green LED light. Viability of D407 cells exposed to green LED light +/+ RB in the presence of RSV remained unchanged comparing to corresponding control. RSV blocked the intracellular ROS production, stimulated or restored the antioxidant enzymes activities and increased the level of reduced GSH in cells exposed to green LED light +/RB. Our data came to support the potential use of RSV as protecting agent of retina’s antioxidant defence system in light-induced stress.