Alessia Fazio

Identification of bioactive constituents of Ziziphus jujube fruit extracts exerting antiproliferative and apoptotic effects in human breast cancer cells

ETHNOPHARMACOLOGICAL RELEVANCE Ziziphus extracts have been used in Traditional Chinese Medicine for the treatment of cancer. AIM OF THE STUDY In the present study we have investigated the effects of Ziziphus jujube extracts (ZEs) on breast cancer. MATERIALS AND METHODS We evaluated the effects of increasing concentrations of ZEs on ERα positive MCF-7 and ERα negative SKBR3 breast cancer cell proliferation using MTT assays. Apoptosis was analyzed by evaluating the involvement of some pro-apoptotic proteins, including Bax, Bad, Bid and PARP cleavage by immunoblotting analysis. Moreover, the effects of ZEs treatment on apoptosis were tested by both DNA fragmentation and terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) staining. By using chromatographic techniques, we identified the constituents of the effective extracts. RESULTS ZE1, ZE2, and ZE4 exerted significant antiproliferative effects on estrogen receptor alpha (ERα) positive MCF-7 (IC(50) values of 14.42, 7.64, 1.69μg/mL) and ERα negative SKBR3 (IC(50) values of 14.06, 6.21, 3.70μg/mL) human breast cancer cells. Remarkably, ZEs did not affect cell viability of both normal human fibroblasts BJ1-hTERT and nonmalignant breast epithelial MCF-10A cells. Treatment with ZEs induced cell death by apoptosis in both malignant breast cells. We found that the most effective extracts ZE2 and ZE4 shared a number of triterpenic acids, already known for their anticancer activities. CONCLUSIONS Our data provide a rational base for the use of Ziziphus extracts in the treatment of breast cancer in Traditional Chinese Medicine.

Comparative analyses of seeds of wild fruits of Rubus and Sambucus species from Southern Italy: Fatty acid composition of the oil, total phenolic content, antioxidant and anti-inflammatory properties of the methanolic extracts

Fruit seeds are byproducts from fruit processing. Characterisation of the bioactive compounds present in seeds and evaluation of their potential biological properties is therefore of particular importance in view of a possible valorisation of seeds as a source of health beneficial components. In this work, we have analysed the seeds of Sambucus and Rubus species in order to identify their bioactive components and to determine the antioxidant and anti-inflammatory activities of the extracts. We first analysed their oil content, in order to assess the fatty acid profile and tocopherol content. Moreover, the methanolic extracts of the seeds were analysed for their total phenolic contents and antioxidant capacities. Polyphenols were identified by HPLC-ESI-MS/MS analysis. Furthermore, extracts were evaluated for their inhibitory effects on the production of LPS-induced inflammatory mediators (NO, CCL-20) in RAW 264.7 cells. Our findings show that the methanolic extracts from Rubus seeds have strong antioxidant and anti-inflammatory properties and could therefore represent an attractive source of bioactive compounds for food, cosmetic, or pharmaceutical applications.

Synthesis of 3,4-bis[(methoxycarbonyl)methyl]thiophene and bis-, ter- and pentathiophenes with alternating 3,4-bis[(methoxycarbonyl)methyl]-substituted rings

Abstract The synthesis of 3,4-bis[(methoxycarbonyl)methyl]thiophene and bis-, ter- and pentathiophenes, with alternating 3,4-bis(methoxycarbonyl)methyl-substituted rings, is reported. These new thiophene derivatives are possible precursors for the preparation of new conducting polymers useful as materials for electronics.

Palladium-catalyzed cycloisomerization of (Z)-(2-en-4-ynyl)amines: a new synthesis of substituted pyrroles

(Z)-(2-En-4-ynyl)amines 1 bearing an internal triple bond undergo smooth cycloisomerization into pyrroles 2 in the presence of catalytic amounts of PdCl2 in conjunction with KCl at 25–100°C in anhydrous N,N-dimethylacetamide. When the triple bond is terminal, spontaneous uncatalyzed cyclization to the corresponding pyrroles takes place.

A new soluble poly(bithiophene)-co-3,4-di(methoxycarbonyl)methyl thiophene for LED

The regioregular polyalkylthiophene reported in this contribution has been electrochemically synthesised starting from a 3,4-bis-methoxyacetyl-terthiophene and carefully characterised. Photoluminescence analysis gave, both in solution and in films, very appreciable quantum yield values and LED devices realised with Ca cathode provided a value of red electroluminescence comparable with those of regioregular poly(3-alkylthiophene)s in the monolayer active material architecture, while if a blend with poly-N-vinylcarbazole and 2-(4-t-butyl-phenyl)-5-(4-biphenyl)-1,3,4-oxadiazole constitutes the active layer a white emission is observed.

Essential oil composition of Citrus medica L. Cv. Diamante (Diamante citron) determined after using different extraction methods

In this paper, we report the qualitative and quantitative composition of the volatile and the oxygenated heterocyclic fraction of citron (Citrus medica L. cv. Diamante (Diamante citron)). The fruits selected for the extraction were of three types: green citron of little size, green citron of big size, and yellow citron after 1 month from the harvest. The essential oils were extracted using three different methods. The nine samples of oil thus obtained were analyzed by high resolution GC (HRGC)-flame ionization detection (FID), HRGC-MS, and RP-HPLC. They differ only in the quantitative composition, while the qualitative profile was the same. The volatile fraction of every sample of oil is characterized by a high content of limonene, gamma-terpinene, and monoterpene aldehydes and a lower content of alpha- and beta-pinene and myrcene, sesquiterpenes, and aliphatic aldehydes. Enantioselective (Es)-GC analysis of the extracts allowed the determination of the enantiomeric distribution of five terpenoid compounds; a prevalence of four dextrorotatory isomers was observed. Oxypeucedanin was the main component of the oxygenated heterocyclic fraction in the extracts of green fruits, while citropten was the major oxygenated compound in the oil obtained from yellow citron.

Isotope dilution method for the assay of rotenone in olive oil and river waters by liquid chromatography/multiple reaction monitoring tandem mass spectrometry

A new approach for the assay of rotenone (1) by isotope dilution mass spectrometry is presented. The extreme toxicity of rotenone, a natural phytodrug with insecticidal and piscicidal activity, recently banned by national and international environmental protection agencies, calls for the development of sensitive and accurate methods of analyses. Accordingly, the proposed protocol is based on the availability of the labeled internal standard rotenone-d3 (3) which can be conveniently prepared by consecutive and specific mono-O-demethylation, and remethylation with methyl iodide-d3. The sensitivity of the method is confirmed by the very low limit of detection (LOD) and quantitation (LOQ) values achieved in the assay of 1 in two distinct fortified matrices, and is further supported by the observed accuracy values.

Unprecedented carbon dioxide effect on a Pd-catalysed oxidative carbonylation reaction: a new synthesis of pyrrole-2-acetic esters

It has been found that carbon dioxide effectively promotes the Pd-catalysed oxidative cyclisation–alkoxycarbonylation of (Z)-(2-en-4-ynyl)amines 1 leading to pyrrol-2-acetic esters 2.

Selective action of human sera differing in fatty acids and cholesterol content on in vitro gene expression

Serum constituents might directly affect metabolic diseases pathogenesis and are commonly used as diagnostic tool. The aim of this study was to investigate the human serum effect on in vitro gene expression, related to nutrients action and involved in lipid metabolism. In detail, 40 human sera were firstly analyzed in fatty acids profile by gas‐chromatography. Then samples were tested through direct addition within culture medium on Hep G2 human hepatoma cells, comparing samples from hypercholesterolemic (average 273 mg/dl) versus normocholesterolemic male subjects (average 155 mg/dl), since this condition is a relevant disease risk factor and is typically consequent to nutritional style. Hypercholesterolemic sera produced a 0.4‐fold reduction of sterol regulatory element binding protein 1c (SREBP‐1c) mRNA (P < 0.05) and a 1.5‐fold increase of UDP‐glucuronosyltransferase 1A1 (UGT1A1) mRNA (P < 0.01). Samples with higher concentrations of n‐6 fatty acids produced a higher expression of UGT1A1 mRNA. Total fatty acids [docosahexaenoic, eicosopentanoic, arachidonic, linolenic, and linoleic acid (DHA, EPA, AA, LNA, and LA, respectively)] in each serum resulted roughly inverse with trend of SREBP‐1c mRNA expression. Serum AA, LA, and trans fatty acids were more abundant in hypercholesterolemic subjects (P < 0.01) while DHA as quota of detected fatty acids was significantly higher in normocholesterolemic subjects (P < 0.05). While it is not possible to indicate which component was responsible for the observed gene modulations, our data indicate that sera differing in lipid profiles, mainly associated with dietary behavior, differentially affect gene expression known to be involved in metabolic and nutritional related conditions. J. Cell. Biochem. 113: 815–823, 2012.

Detection of ochratoxin A based on the use of its diastereoisomer as an internal standard

The excellent separation achieved by HPLC of the diastereoisomers 3-(R) and 3-(S) of ochratoxin A and their identification by fluorimetric detection has prompted the development of a new method for the detection of ochratoxin A, one of the most abundant food-contaminating mycotoxins in the world. The new method is based on the use of N-[(5-chloro-3,4-dihydro-8-hydroxy-3-(S)-methyl-1-oxo-1H-2-benzopyran-7-yl)carbonyl]-L-phenylalanine as an internal standard. This compound was synthesized by a coupling reaction between commercially available phenylalanine and ochratoxin α. The effectiveness of the method is confirmed by the very low limit of detection and quantitation values achieved for the detection of ochratoxin A in two fortified matrixes (0.002 and 0.005 μg L−1, respectively, in wine; 0.029 and 0.072 μg kg−1, respectively, in flour) and is further supported by the observed accuracy values. The new approach meets the requirement for the detection of subpicogram amounts of contaminating toxins in complex matrices such as foods. Moreover, the method is affordable to any laboratory because it is based on relatively inexpensive separation and detection methodologies.