Determination of organic acids in plants of Silene paradoxa L. by HPLC

According to the general behavior that organic acids steadily bind metals, a specific and highly reproducible HPLC separation method with photodiode array detection has been improved for their determination and quantification in biological materials. The separation was carried out on an Alltima C-18 reverse phase column. The mobile phase was 125 mM KH2PO4, adjusted to pH 2.5 with concentrated H3PO4, and optimum separation efficiency was obtained by using a 2 mL min(-1) flow rate. Detection wavelength for quantitative measurement was 210 nm. The run time of each sample was 20 min, with a spectra collection frequency of 5 spectra s(-1). Organic acids were identified by comparing the retention times of the samples against retention times of the standards and confirmed with spectral (190-700 nm) signature. Because organic acids could steadily bind metals in plant tissues and due to the strong matrix effect observed, the addition method was applied for quantitative analysis and its performance evaluated.     

Protective effect of vanilloids against tert-butyl hydroperoxide-induced oxidative stress in vero cells culture

This study investigated the effect of synthetic capsiate, a simplified analogue of capsiate, and vanillyl alcohol on the oxidative stress induced by tert-butyl hydroperoxide (TBH) in a line of fibroblasts derived from monkey kidney (Vero cells). In response to the TBH-mediated oxidative stress, a reduction of the levels of total unsaturated fatty acids and cholesterol was observed, and a rise in the concentrations of conjugated dienes fatty acids hydroperoxides and 7-ketocholesterol. Pretreatment with both synthetic capsiate and vanillyl alcohol preserved Vero cells from oxidative damage and showed a remarkable protective effect on the reduction of the levels of total unsaturated fatty acids and cholesterol, inhibiting the increase of MDA, conjugated dienes fatty acids hydroperoxides, and 7-ketocholesterol. Both compounds were effective against peroxidation of cell membrane lipids induced by TBH, with synthetic capsiate essentially acting as a pro-drug of vanillyl alcohol, its hydrophilic hydrolytic derivative.     

Identification and quantification of phenolic compounds in grapes by HPLC-PDA-ESI-MS on a semimicro separation scale

Reversed phase high performance liquid chromatography (RP-HPLC) on a semimicro separation scale was employed to develop a straightforward method for the simultaneous separation, identification, and quantification of phenolic compounds occurring in whole berries of Vitis vinifera, which comprise phenolic acids, flavonols, catechins, stilbenes, and anthocyanins. A C-18 narrow bore column of 150 x 2.0 mm I.D. and a semimicro photodiode array detector (PDA) cell of 2.5 microL, in conjunction with a mass spectrometry detector equipped with an electrospray ionization source (ESI-MS) to confirm peak identification, were employed. The C-18 narrow bore column was eluted by a multisegment gradient of increasing concentration of acetonitrile in water-formic acid solution that was optimized on the basis of the results of a study carried out to evaluate the influence of mobile phase composition and gradient shape on separation performance and detection sensitivity by ESI-MS. The identification of individual phenolic compounds was performed on the basis of their retention times and both UV-visible and mass spectra, acquired by a mass spectrometer (MS) equipped with an electrospray ionization (ESI) source, employed in conjunction with the PDA detector. Libraries comprising retention times, UV-visible, and mass spectra for major phenolic compounds expected in grape berries were made by subjecting solutions of each phenolic standard to the optimized RP-HPLC method. Quantification of individual compounds was performed by the external standard method using a six point regression graph of the UV-visible absorption data collected at the wavelength of maximum absorbance of each analyte determined by the PDA spectra. The RP-HPLC method was validated in terms of linearity of calibration graphs, limits of detection, limits of quantification, repeatability, and accuracy, which was evaluated by a recovery study. The developed method was successfully applied to identify the phenolic compounds occurring in the whole berries of nine red and one white grape of different varieties of Vitis vinifera, comprising some autochthonous varieties of south Italy such as Aglianico, Malvasia Nera, Uva di Troia, Negroamaro, Primitivo, and Susumaniello. Large differences in the content of phenolic compounds was found in the investigated grape varieties. As expected, only glycosilated flavonols were quantified, and the total amount of these compounds was higher in the whole berries of red grapes than in the white Moscato, where the most abundant phenolic compound was quercetin 3-O-glucoside. In almost all samples, the most and least abundant anthocyanins were malvidin 3-O-glucoside and cyanidin 3-O-glucoside, respectively, with the exception of Uva di Troia where the least abundant anthocyanin was delphinidin 3-O-glucoside.     

Pollution Bioindicators: Statistical Analysis of a Case Study

In this paper a three-step procedure is proposed to deal with ecological data, usually very complex in their treatment. The three steps – exploratory, confirmatory, and modelling phases – reflect the different methodological approaches necessary in each phase of the study. To illustrate the methodology, a case study is proposed, concerning the suitability of plants as pollution bioindicators. Samples of differently aged Pinus pinea L. needles were collected throughout 1 year in three different locations, whose human disturbance was known to be different. In the samples some morphological and functional parameters were measured, whose relation with the stress was already known. The exploratory analysis suggested pollution with human origin, the needle’s age, and the environmental conditions as the main factors of influence of damage. The confirmatory analysis confirmed both site and age as main factors and occasionally the sampling date. On this basis, some models were estimated separately for each site: models that best described the damage as function of age resulted non-linear and some of them with seasonal fluctuations. As a result, whereas the models described well enough the pollution temporal variation, the difference of pollution in the sites was best described by the different values of the models parameters in the different sites. In short, different pollution conditions are described better by the damage trend than by the individual measures. The three-step procedure resulted of high utility in outlining the most interesting relations to investigate through the modelling, the opportunity to model the indicators variation along time separately for each site, and to introduce the seasonal variation in some models.     

Decreased levels of metalloproteinase-9 and angiogenic factors in skin lesions of patients with psoriatic arthritis after therapy with anti-TNF-α

Background

Inflammation represents an early and key event in the development of both the cutaneous psoriasis and psoriatic arthritis. Compelling evidences indicate that the production of TNF-α plays a central role in psoriasis by sustaining the inflammatory process in the skin as well as in the joints. Among the multiple effects produced by TNF-α on keratinocytes, the induction of matrix metalloproteinase-9 (MMP-9), a collagenase implicated in joint inflammatory arthritis which acts as an angiogenesis promoting factor, might represent a key mechanism in the pathogenesis of the disease. Aims of the present study were to investigate a) the role of MMP-9 in the development of psoriasis by assessing the presence of MMP-9 in lesional skin and in sera of psoriatic patients; b) the association of MMP-9 with the activity of the disease; c) the relationship between MMP-9 and TNF-α production.

Methods

Eleven psoriatic patients, clinically presenting joint symptoms associated to the cutaneous disease, were included in a therapeutic protocol based on the administration of anti-TNF-α monoclonal antibody (Infliximab). Sera and skin biopsies were collected before treatment and after 6 weeks of therapy. Tissues were kept in short term cultures and production soluble mediators such as TNF-α, MMP-9, MMP-2, VEGF and E-Selectin, which include angiogenic molecules associated to the development of plaque psoriasis, were measured in the culture supernatants by immunoenzymatic assays (ng/ml or pg/ml per mg of tissue). MMP-9 concentrations were also measured in the sera. The cutaneous activity of disease was evaluated by the Psoriasis Area and Severity Index (PASI).

Results

Clinical and laboratory assessment indicated that all but one patients had a significant improvement of the PASI score after three months of therapy. The clinical amelioration was associated to a significant decrease of MMP-9 (P = 0.017), TNF-α (P = 0.005) and E-selectin (P = 0.018) levels, spontaneously released by lesional biopsies before and after therapy. In addition, significant correlations were found between the PASI measurements and TNF-α (r² = 0.33, P = 0.005), MMP-9 (r² = 0.25, P = 0.017), E-selectin (r² = 0.24, P = 0.018) production. MMP-9 levels were significantly correlated with those of TNF-α (r² = 0.30, P = 0.008). A significant decrease of MMP-9 in the sera, associated to the clinical improvement was also found.

Conclusion

Our findings show the existence of a direct relationship between MMP-9 and TNF-α production strongly suggesting that MMP-9 may play a key role in the skin inflammatory process in psoriasis.

     

Lipoxygenase involvement in ripening strawberry

The enzymatic activity, subcellular localization, and immunolocalization of plant lipoxygenase (LOX) in strawberry fruits (Fragaria x ananassa, Duch) were investigated. Chemical and enzymatic properties of LOX have been characterized, and the LOX capability of oxygenating free and esterified unsaturated fatty acids into C6 volatile aldehydes has been confirmed. Fruits at unripe, turning, and ripe stages were analyzed for LOX activity and protein localization by Western blots, two-dimensional electrophoresis, and immunolocalization analyses. The ability of strawberry tissues to in vivo metabolize linolenic acid or linoleic acid into C6 volatile aldehydes and the LOX products was also analyzed. Analysis of strawberry proteins showed that a number of LOX forms, corresponding to at least two mobility groups of approximately 100 and 98 kDa and pI values ranging between 4.4 and 6.5, were present. Confocal and electron microscopy analyses support the idea that LOX proteins are associated to lipid-protein aggregates. Both exogenously supplied linoleate and linolenate were converted into hexanal and trans-2-hexenal at the three fruit-ripening stages. Our experiments suggest the presence of different LOX isoforms in strawberry fruits and that the lipoxygenase-hydroperoxide lyase pathway plays a role in converting lipids to C6 volatiles during ripening.     

Dual Targeting of PTP1B and Aldose Reductase with Marine Drug Phosphoeleganin: A Promising Strategy for Treatment of Type 2 Diabetes

An in-depth study on the inhibitory mechanism on protein tyrosine phosphatase 1B (PTP1B) and aldose reductase (AR) enzymes, including analysis of the insulin signalling pathway, of phosphoeleganin, a marine-derived phosphorylated polyketide, was achieved. Phosphoeleganin was demonstrated to inhibit both enzymes, acting respectively as a pure non-competitive inhibitor of PTP1B and a mixed-type inhibitor of AR. In addition, in silico docking analyses to evaluate the interaction mode of phosphoeleganin with both enzymes were performed. Interestingly, this study showed that phosphoeleganin is the first example of a dual inhibitor polyketide extracted from a marine invertebrate, and it could be used as a versatile scaffold structure for the synthesis of new designed multiple ligands.     

An integrated approach for cetacean knowledge and conservation in the central Mediterranean Sea using research and social media data sources

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Site fidelity and movement patterns of short‐finned pilot whales within the Canary Islands: Evidence for resident and transient populations

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Potential effects of global climate change on fisheries in the Trasimeno Lake (Italy), with special reference to the goldfish Carassius auratus invasion and the endemic southern pike Esox cisalpinus decline

Global climate changes have led to a gradual warming of the planet, resulting in decreased precipitation and rising temperatures in Mediterranean inland waters. In Trasimeno Lake, the largest shallow lake in Italy, some non‐native fish species have probably benefited from these changes as they are thermophilic and characterised by wider habitat preferences. Fish data collected by gillnets and fyke nets between 1956 and 2016, and by electrofishing in 1993 and 2014, were used to analyse changes over time in the fish community in relation to environmental conditions. An explosion in goldfish Carassius auratus (L.), following its introduction in 1988, coupled with water level fluctuations and reduced transparency, contributed to the reduction in commercial fish catch in the lake, and to the decline of the endemic southern pike Esox cisalpinus Bianco & Delmastro, already threatened by reduced spawning habitat and interspecific competition with other non‐native predatory fishes.