Peptidomic approach for the identification of peptides with potential antioxidant and anti-hyperthensive effects derived from Asparagus by-products

Asparagus waste represents products of great interest since many compounds with high biological value are located in the lower portion of the spears. The extraction of bioactive compounds from asparagus by-products is therefore crucial for the purpose of adding value to these by-products. In this paper, bioactive peptides from asparagus waste were extracted, digested, purified and identified. In particular, Alcalase® was chosen as the enzyme to use to obtain protein hydrolysate due to its low cost and, consequently, the possibility of implementing the method on a large scale. In order to simplify the peptide extract to reach better identification, the hydrolysate was fractionated by reversed-phase chromatography in 10 fractions. Two tests were carried out for antioxidant activity (ABTS-DPPH) and one for antihypertensive activity (ACE). Fractions with a higher bioactivity score were identified by peptidomics technologies and screened for bioactivity with the use of bioinformatics. For ACE-inhibitor activity, two peptides were synthetized, PDWFLLL and ASQSIWLPGWL, which provided an EC50 value of 1.76 µmol L-1 and 4.02 µmol L-1, respectively. For the antioxidant activity, by DPPH assay, MLLFPM exhibited the lowest EC50 value at 4.14 µmol L-1, followed by FIARNFLLGW and FAPVPFDF with EC50 values of 6.76 µmol L-1 and 10.01 µmol L-1, respectively. A validation of the five identified peptides was also carried out. The obtained results showed that peptides obtained from asparagus by-products are of interest for their biological activity and are suitable for being used as functional ingredients

Pharmacoproteomic characterisation of human colon and rectal cancer

Most molecular cancer therapies act on protein targets but data on the proteome status of patients and cellular models for proteome-guided pre-clinical drug sensitivity studies are only beginning to emerge. Here, we profiled the proteomes of 65 colorectal cancer (CRC) cell lines to a depth of > 10,000 proteins using mass spectrometry. Integration with proteomes of 90 CRC patients and matched transcriptomics data defined integrated CRC subtypes, highlighting cell lines representative of each tumour subtype. Modelling the responses of 52 CRC cell lines to 577 drugs as a function of proteome profiles enabled predicting drug sensitivity for cell lines and patients. Among many novel associations, MERTK was identified as a predictive marker for resistance towards MEK1/2 inhibitors and immunohistochemistry of 1,074 CRC tumours confirmed MERTK as a prognostic survival marker. We provide the proteomic and pharmacological data as a resource to the community to, for example, facilitate the design of innovative prospective clinical trials. © 2017 The Authors. Published under the terms of the CC BY 4.0 licens

Characterization of acrosin and acrosin binding protein as novel CRISP2 interacting proteins in boar spermatozoa

Background: Previously, we reported that cysteine-rich secretory protein 2 is involved in high molecular weight complexes in boar spermatozoa. These cysteine-rich secretory protein 2protein complexes are formed at the last phase of sperm formation in the testis and play a role in sperm shaping and functioning. Objectives: This study aimed to identify cysteine-rich secretory protein 2 interacting partners. These binding partner interactions were investigated under different conditions, namely, non-capacitating conditions, after the induction of in vitro sperm capacitation and subsequently during an ionophore A23187-induced acrosome reaction. Materials and Methods: The incubated pig sperm samples were subjected to protein extraction. Extracted proteins were subjected to blue native gel electrophoresis and native immunoblots. Immunoreactive gel bands were excised and subjected to liquid chromatography–mass spectrometry (LC-MS) analysis for protein identification. Protein extracts were also subjected to CRISP2 immunoprecipitation and analyzed by LC-MS for protein identification. The most prominent cystein-rich secretory protein 2 interacting proteins that appeared in both independent LC-MS analyses were studied with a functional in situ proximity interaction assay to validate their property to interact with cystein-rich secretory protein 2 in pig sperm. Results: Blue native gel electrophoresis and native immunoblots revealed that cystein-rich secretory protein 2 was present within a ∼150 kDa protein complex under all three conditions. Interrogation of cystein-rich secretory-protein 2-immunoreactive bands from blue native gels as well as cystein-rich secretory protein 2 immunoprecipitated products using mass spectrometry consistently revealed that, beyond cystein-rich secretory protein 2, acrosin and acrosin binding protein were among the most abundant interacting proteins and did interact under all three conditions. Co-immunoprecipitation and immunoblotting indicated that cystein-rich secretory protein 2 interacted with pro-acrosin (∼53 kDa) and Aacrosin binding protein under all three conditions and additionally to acrosin (∼35 kDa) after capacitation and the acrosome reaction. The colocalization of these interacting proteins with cystein-rich secretory protein 2 was assessed via in situ proximity ligation assays. The colocalization signal of cystein-rich secretory protein 2 and acrosin in the acrosome seemed dispersed after capacitation but was consistently present in the sperm tail under all conditions. The fluorescent foci of cystein-rich secretory protein 2 and acrsin binding protein colocalization appeared to be redistributed within the sperm head from the anterior acrosome to the post-acrosomal sheath region upon capacitation. Discussion and Conclusion: These results suggest that CRISP2 may act as a scaffold for protein complex formation and dissociation to ensure the correct positioning of proteins required for the acrosome reaction and zona pellucida penetration

Approaching an investigation of multi-dimensional inequality through the lenses of variety in models of capitalism

After a synthetic presentation of the state of poverty and inequality in the world and the contradictions incurred by economic theory in this field after decades of globalization and in the midst of a persisting global crisis, in paragraphs 2. and 3. we outline the rational for our theoretical analysis, underlining two main aspects. First of all, in paragraph 2. we recall the reasons which makes inequality a multidimensional phenomenon, while in paragraph 3. we explore the reasons why the models of capitalism theory is relevant for studying multidimensional inequality. These paragraphs emphasise that inequality is a multidimensional and cumulative phenomenon and it should not be conceived only as the result of the processes of personal and functional distribution of income and wealth, which even by themselves are intrinsically multidimensional. The basic idea is that institutions, the cobweb of relations among them and their interaction with the economic structure define the model of capitalism which characterises a specific country and this, in turn, affects the level and the dynamics of inequality. This approach is consistent with the sociological approach by Rehbein and Souza (2014), based on the analytical framework developed by Pierre Bourdieu. In paragraph 4. we outline the rational for our empirical analysis, applying the notion of institutional complementarity and examining the relationship between institutional complementarity, models of capitalism and inequality. Besides, refining Amable’s analysis (2003), we provide empirical evidence on the relationship between inequality in income distribution and models of capitalism. Additionally, basing on cluster analysis, we identify six different models of capitalism in a sample of OECD countries, provide preliminary evidence on the different level of inequality which characterises each model and suggest that no evidence supports of the idea that a single model of capitalism is taking shape in this sphere in EU. In paragraph 5. we give some hints about issues in search for a new interpretation capable to fasten together the process of increasing inequality, the notion of symbolic violence and the models of capitalism theory. In the last paragraph we focus on conclusions useful for carrying on our research agenda

Occupazione e disoccupazione in Italia

RIVISTA DI ECONOMIA E POLITICA INDUSTRIAL