Influence of cultivar and concentration of selected phenolic constituents on the in vitro chemiopreventive potential of olive oil extracts

One of the main olive oil phenolic compounds, hydroxytyrosol (3,4-DHPEA), exerts in vitro chemopreventive activities (antiproliferative and pro-apoptotic) on tumor cells through the accumulation of H(2)O(2) in the culture medium. However, the phenol composition of virgin olive oil is complex, and 3,4-DHPEA is present at low concentrations when compared to other secoiridoids. In this study, the in vitro chemopreventive activities of complex virgin olive oil phenolic extracts (VOO-PE, derived from the four Italian cultivars Nocellara del Belice, Coratina, Ogliarola, and Taggiasca) were compared to each other and related to the amount of the single phenolic constituents. A great chemopreventive potential among the different VOO-PE was found following this order: Ogliarola > Coratina > Nocellara > Taggiasca. The antiproliferative and pro-apoptotic activities of VOO-PE were positively correlated to the secoiridoid content and negatively correlated to the concentration of both phenyl alcohols and lignans. All extracts induced H(2)O(2) accumulation in the culture medium, but this phenomenon was not responsible for their pro-apoptotic activity. When tested in a complex mixture, the olive oil phenols exerted a more potent chemopreventive effect compared to the isolated compounds, and this effect could be due either to a synergistic action of components or to any other unidentified extract constituent.     

Phylogeography and conservation perspectives of an endangered macaronesian endemic: Picconia azorica (Tutin) Knobl. (Oleaceae)

The Azores provide an excellent field test for research activities aimed at developing conservation strategies for endangered tree species. In this work, the urgency to promote Picconia azorica conservation programs addressed (a) insights into the biotaxonomy of the species (including an evaluation of its origin and relationships with the only congeneric species of P. azorica: P. excelsa from the Canary and Madeira islands) and (b) the evaluation of P. azorica genetic diversity. Plastid DNA sequence analysis and molecular markers (RFLP and SSR) were used for this purpose. Phylogenetic data suggest the monophyly of Picconia and support a late Miocene divergence of the two species. Three polymorphic cpSSR loci allowed the identification of five different haplotypes in P. azorica. Uniqueness and relictuality of lineages are presented and discussed. Picconia azorica intra-specific diversity patterns revealed low genetic diversity and a weak genetic structure, which could result from long-lasting ecological stability and efficient inter-island seed movement that have been severely affected in recent times. The species survival is at risk, and we suggest management practices focusing on ex situ and in situ conservation units based on eco-genetic data. Additional measures contributing to mild erosion of the genepool and to remove barriers to seed dispersal are indicated.     

Differentiation and characterization of rat adipose tissue mesenchymal stem cells into endothelial‐like cells

In this study, mesenchymal stem cells were isolated from rat adipose tissue (AD‐MSCs) to characterize and differentiate them into endothelial‐like cells. AD‐MSCs were isolated by mechanical and enzymatic treatments, and their identity was verified by colony‐forming units (CFU) test and by differentiation into cells of mesodermal lineages. The endothelial differentiation was induced by plating another aliquot of cells in EGM‐2 medium, enriched with specific endothelial growth factors. Five subcultures were performed. The expression of stemness genes (OCT4, SOX2 and NANOG) was investigated. The presence of CD90 and the absence of the CD45 were evaluated by flow cytometry. The endothelial‐like cells were characterized by the evaluation of morphological changes and gene expression analysis for endothelial markers (CD31, CD144, CD146). Characterization of AD‐MSCs showed their ability to form clones, to differentiate in vitro and the OCT‐4, SOX‐2, NANOG genes expression. Immunophenotypic characterization showed the CD90 presence and the CD45 absence. The endothelial‐like cells showed morphological changes, the expression of CD31, CD144, CD146 genes and the presence of CD31 membrane receptor. Matrigel assay showed their ability to form network and vessels‐like structures. This study lays the foundations for future evaluation of the potential AD‐MSCs pro‐angiogenic and therapeutic role.     

The effects of drought on the water use, fruit development and oil yield from young olive trees

In Marlborough, New Zealand, olives are becoming an important crop alongside grapes. However, despite olives being drought resistant, they are generally planted on the poorer free-draining soils. Also, with the strong increase in cropping area, the demand for irrigation water has increased dramatically. In this research, we investigate the impact of short-term water stress on plant physiological processes, crop yield and oil quality in Marlborough, New Zealand. For that purpose, during the dry summer of 2000–2001, two trees were kept without irrigation for 64 days while two neighbouring trees were irrigated following standard practice. The trees were measured for transpiration (E), leaf and stem water potential (Ψ_L and Ψ_S), every other day, from dawn to dusk for three weeks from just before irrigation was started up again. All four trees were wired up for measuring stem sap flow (T) which was recorded hourly and a basic meteorological station provided weather data. Fruit and shoot development was measured weekly. It was found that under the short period of dry conditions with soil moisture (() dropping to <5%, olive trees kept functioning at a very low level with Ψ_L and Ψ_S reduced from −1 to <−4.0 MPa (T) reduced from 20 to 5 mm/h and (E) reduced from 1.5 to 1.0 mmol m⁻² s⁻¹. Within 10 days of restarting irrigation all these parameters were back to pre-drought levels. Both fruit and shoot growth came to a standstill within a week after drought was induced. During the first few days after re-watering, a high variability in Ψ_L was found between leaves from the same trees. This variability disappeared after six days. Shoot growth did not recover after re-watering but fruit growth rate, became the same as for continuously irrigated trees within days, but fruit size did not manage to recover before harvest. Yield from the dry trees was low because berry and pit weight were reduced by almost 50% at harvest, had a lower oil and percentage and were lower in phenolics. Stem sap flow was found to give a very good continuous measurement for the hydration status of the olive trees.     

Shade effect on photosynthesis and photoinhibition in olive during drought and rewatering

Olive tree (Olea europaea L.) is commonly grown under environmental conditions characterised by water deficit, high temperatures and irradiance levels typical of Mediterranean semi-arid regions. Measurement of gas exchange, chlorophyll content, chlorophyll fluorescence and photoinhibition was carried out on two-year-old olive trees (cv. ‘Coratina’) subjected to a 21-day period of water deficit followed by 23 days of rewatering. At the beginning of the experiment, plants were divided in to two groups and subjected to different light regimes: exposed plants (EP) under a mean photosynthetically active radiation (PAR) at mid-day of 1800 μmol m⁻² s⁻¹ and shaded plants (SP) under a mean PAR of 1200 μmol m⁻² s⁻¹. The effect of drought and high irradiance levels caused a reduction of gas exchange and photosystem 2 (PSII) efficiency, in terms of quantum yield of PSII (Φ_PSII) both in EP and SP. Shading conditions allowed plants to maintain a high photosynthetic activity at low values of stomatal conductance, whereas in EP the reductions in photosynthetic efficiency and intrinsic water efficiency were due to non-stomatal components of photosynthesis. The decrease in photosynthetic activity and the increase of photoinhibition under drought were more marked in EP than in SP. Full sunlight caused in EP a higher non-photochemical quenching, whereas SP showed a better photochemical efficiency. The information here obtained can be important to understand the mechanisms by which olive plants can minimize photoinhibition when subjected to simultaneous abiotic stresses.     

PDGFs and PDGFRs in canine osteosarcoma: New targets for innovative therapeutic strategies in comparative oncology

Platelet derived growth factor receptor (PDGFR)α and PDGFRβ are tyrosine kinase receptors that are overexpressed in 70–80% of human osteosarcomas (OSAs) and may be suitable therapeutic targets for specific kinase inhibitors (TKIs). Canine OSA shows histopathological and clinical features similar to human OSA, and is considered an excellent model in comparative oncology. This study investigated PDGF-A, PDGF-B, PDGFRα and PDGFRβ expression in 33 canine OSA samples by immunohistochemistry and in seven primary canine OSA cell lines by Western blot and quantitative PCR analysis.Immunohistochemical data showed that PDGF-A and PDGF-B are expressed in 42% and 60% of the OSAs analysed, respectively, while PDGFRα and PDGFRβ were expressed in 78% and 81% of cases, respectively. Quantitative PCR data showed that all canine OSA cell lines overexpressed PDGFRα, while 6/7 overexpressed PDGFRβ and PDGF-A relative to a normal osteoblastic cell line. Moreover, in vitro treatment with a specific PDGFR inhibitor, AG1296, caused a dose- and time-dependent decrease in AKT phosphorylation. Collectively, these data show that PDGFRs/PDGFs are co-expressed in canine osteosarcomas, which suggests that an autocrine and/or paracrine loop is involved and that they play an important role in the aetiology of OSA. PDGFRs may be suitable targets for the treatment of canine OSA with a specific TKI.