Enzo Perri

A consensus list of microsatellite markers for olive genotyping

Cultivar identification is a primary concern for olive growers, breeders, and scientists. This study was aimed at examining the SSR markers retrieved from the literature and currently used in olive study, in order to select those most effective in characterizing the olive accessions and to make possible the comparison of data obtained by different laboratories. Olive microsatellite profiles were assessed by four independent laboratories, which analyzed 37 pre-selected SSR loci on a set of 21 cultivars. These SSR markers were initially tested for their reproducibility, power of discrimination and number of amplified loci/alleles. Independent segregation was tested for each pair of SSRs in a controlled cross and the allelic error rate was quantified. Some of them were finally selected as the most informative and reliable. Most of the alleles were sequenced and their sizes were determined. Profiles of the reference cultivars and a list of alleles with their sizes obtained by sequencing are reported. Several genetic parameters have been analysed on a larger set of cultivars allowing for a deeper characterization of the selected loci. Results of this study provide a list of recommended markers and protocols for olive genotyping as well as the allelic profile of a set of reference cultivars that would be useful for the establishment of a universal database of olive accessions.

Effects of olive oil polyphenols on fatty acid synthase gene expression and activity in human colorectal cancer cells

Oleuropein (OL) and hydroxytyrosol (HT), the main olive oil polyphenols, possess anti-proliferative effects in vitro. Fatty acid synthase, a key anabolic enzyme of biosynthesis of fatty acids, plays an important role in colon carcinoma development. Our aim was to investigate whether gene expression of FAS, as well as its enzymatic activity, is regulated by HT and OL in two human colon cancer cell lines, as HT-29 and SW620. In addition, we investigated the effects of these polyphenols on growth and apoptosis in these cells. FAS gene expression and activity in treated HT-29 and SW620 cells were evaluated by real-time PCR and radiochemical assay, respectively. Cell growth and apoptosis, after polyphenols treatment, were measured by MTT test and flow cytometry, respectively. The inhibition of proliferation, detected after HT treatment, was mediated by an inhibition of FAS expression and its enzymatic activity in SW620 cells, while the anti-proliferative effect in HT-29 cells seems to be independent from FAS. OL exerted an anti-proliferative effect only on SW620 cells with a mechanism which excluded FAS. Olive oil polyphenols used were able to induce apoptosis in both cell lines studied. The increase of apoptosis in these cells was accompanied by the block of cell cycle in the S phase. This study demonstrates that HT and OL may induce anti-proliferative and pro-apoptotic effects only in certain human colorectal cancer cell types. These effects are FAS mediated only in SW620 cells after treatment with HT.

Influence of Olive Maturity Stage and Geographical Origin on Some Minor Components in Virgin Olive Oil of the Chemlali Variety

Minor compounds such as sterols, aliphatic alcohols, tocopherols, and chlorophylls in virgin olive oil from Chemlali cultivar were analyzed during the maturity process. The study concerns oils from the following three different olive growing areas of Tunisia: Sfax, Sidi Bouzid, and Enfidha. Analytical results showed that both the maturity process and the olive provenances influence the evolution of the content of these compounds.

Virtual freezing of the hemiacetal–aldehyde equilibrium of the aglycones of oleuropein and ligstroside present in olive oils from Carolea and Coratina cultivars by ionspray ionization tandem mass spectrometry

The composition of the equilibrium between aldehydes and hemiacetals was evaluated by ionspray ionization tandem mass spectrometry. The methanolic extracts of virgin olive oil from Carolea and Coratina cultivars contain a complex mixture of hydroxytyrosol (3) and tyrosol (4) derivatives (5, 6) characterized by the presence of aldehyde moieties in equilibrium with their hydrated form (8) or with the corresponding methanol hemiacetals 9 and 10. The equilibrium was frozen by droplet evaporation, thus allowing the structure determination of each component. The formation of the decarboxylated species 11-14, reported previously, was not observed.

Direct Identification of Phenolic Glucosides from Olive Leaf Extracts by Atmospheric Pressure Ionization Tandem Mass Spectrometry

Pneumatically assisted electrospray (or ionspray) coupled with liquid chromatography was applied to the identification of the phenolic glucoside content of olive leaf directly from the crude extracts. The mass spectra of the positive ions provide insights into the composition of the phenolic constituents. Oleuropein, ligstroside and a disaccharide containing the hydroxytyrosol moiety were found in olive leaf of Olea europea L. cv. Cassanese and their structures were thoroughly determined by tandem mass spectrometry.

Study of oils from Calabrian olive cultivars by chemometric methods

Abstract A study of characterisation of a typical Italian food is performed by chemometrics. The olive oils from some cultivars of Calabria have been characterised according to their origin and olive genotype using the chemical information mainly provided by 14 chemical parameters of virgin olive oils. In particular, the models of three cultivars of Calabria (Carolea, Cassanese, Dolce di Rossano) were deeply studied. The microclimate of this region has a lower influence on fatty acid composition than the genotype. Using simple and relatively inexpensive analytical parameters as fatty acids and chemometric techniques it has been possible to characterise and classify the olive oils (60–94% prediction rate).

Characterization of cassanese olive cultivar through the identification of new trace components by ionspray tandem mass spectrometry

Pneumatically assisted electrospray (ionspray) coupled with liquid chromatography was applied in the identification of antioxidants present in traces in the leaves of Olea europea L. cv. Cassanese, a typical olive tree from the northeastern part of Calabria region in Italy. The structures of these potential biomarkers were evaluated with reference to established mass spectrometric rules or, in one case, by matching the tandem mass spectrum of the unknown with that of an authentic sample obtained by synthesis. Copyright

The effects of a polyphenol present in olive oil, oleuropein aglycone, in an experimental model of spinal cord injury in mice.

Several olive oil phenolic compounds, such us oleuropein have attracted considerable attention because of their antioxidant activity, anti-atherosclerotic and anti-inflammatory properties. The aim of this experimental study was to determine the effect of oleuropein aglycone, a hydrolysis product of oleuropein, in the inflammatory response, in particular in the secondary injury associated with the mouse model of spinal cord trauma. The injury was induced by application of vascular clips to the dura via a four-level T5-T8 laminectomy in mice. Oleuropein aglycone was administered in mice (100 μg/kg, 40 μg/kg, 20 μg/kg, 10% ethanol, i.p.) 1h and 6h after the trauma. The treatment with oleuropein aglycone significantly decreased: (1) histological damage, (2) motor recovery, (3) nuclear factor (NF)-κB expression and IKB-α degradation, (4) protein kinase A (PKA) activity and expression, (5) pro-inflammatory cytokines production such as tumor necrosis factor (TNF-α) and interleukin-1β (IL-1β), 6) inducible nitric oxide synthase (iNOS) expression, (7) neutrophil infiltration, (8) lipid peroxidation, (9) nitrotyrosine and poly-ADP-ribose (PAR) formation, (10) glial cell-derived neurotrophic factor (GDNF) levels, (11) apoptosis (TUNEL staining, FAS ligand expression, Caspase 3, Bax and Bcl-2 expression). Thus, we propose that olive oil phenolic constituents such as oleuropein aglycone may be useful in the treatment of various inflammatory diseases.

Qualitative evaluation of olive oils from new olive selections and effects of genotype and environment on oil quality

In the dynamically changing olive oil industry, new cultivars with greateradaptability to modern cul - tivation techniques and with high quality oils are needed. In this paper, oil quality, in terms of fatty acid compo - sition and content in phenolic compounds, was analysed for many new genotypes previously selected in a bree- ding program and cultivated in three different locations of central and southern Italy. Data on some cultivars commonly grown in each area were included for comparison. The availability of data from many genotypes cul - tivated in all three locations allowed quantitative analyses of the genetic and environmental effects on the oil qua - lity traits studied. The acidic composition varied greatly both with genotype and with environment and so did the concentration in phenols, though the effect of genotype on phenols was not significant (P=0.09). The fatty acid composition, particularly the oleic/(palmitic+linoleic) ratio, appeared predominantly under genetic control, with a broad sense heritability of 0.58 while the environmental effect explained 0.31 of the total variance. The oil con - tent in phenolic compounds, instead, had lower heritability (0.29) and was more affected by the environment, which explained 0.50 of the total variance. Few genotypes were selected as the best for each location, but none performed best in all locations. This suggests that, in olive, the highest oil quality is difficult to achieve with a sin - gle genotype in different environments, due to a strong or even predominant effect of the environment on some quality traits. More likely, combinations of genotypes and territories can produce oils with high and typical qua - lity. In this paper, we provide information on potential new genotypes with promising performances in different Italian regions. Further testing of oil quality, together with agronomic performance, is necessary prior to any release of new cultivars.

Effects of a polyphenol present in olive oil, oleuropein aglycone, in a murine model of intestinal ischemia/reperfusion injury.

Dietary olive oil supplementation and more recently, olive oil phenols have been recommended as important therapeutic interventions in preventive medicine. Ole has several pharmacological properties, including antioxidant, anti‐inflammatory, antiatherogenic, anticancer, antimicrobial, and antiviral and for these reasons, is becoming an important subject of study in recent years. The aim of this study was to investigate the effects of Ole aglycone on the modulation of the secondary events in mice subjected to intestinal IRI. This was induced in mice by clamping the superior mesenteric artery and the celiac trunk for 30 min, followed by release of the clamp, allowing reperfusion for 1 h. After 60 min of reperfusion, animals were killed for histological examination of the ileum tissue and immunohistochemical localization of proinflammatory cytokines (TNF‐α and IL‐1β) and adhesion molecules (ICAM‐1 and P‐sel); moreover, by Western blot analysis, we investigated the activation of NF‐κB and IκBα. In addition, we evaluated the apoptosis process, as shown by TUNEL staining and Bax/Bcl‐2 expressions. The results obtained by the histological and molecular examinations showed in Ole aglycone‐treated mice, a decrease of inflammation and apoptosis pathway versus SAO‐shocked mice. In conclusion, we propose that the olive oil compounds, in particular, the Ole aglycone, could represent a possible treatment against secondary events of intestinal IRI.