Effects of Human and Porcine Adipose Extracellular Matrices Decellularized by Enzymatic or Chemical Methods on Macrophage Polarization and Immunocompetence

The decellularized extracellular matrix (ECM) obtained from human and porcine adipose tissue (AT) is currently used to prepare regenerative medicine bio-scaffolds. However, the influence of these natural biomaterials on host immune response is not yet deeply understood. Since macrophages play a key role in the inflammation/healing processes due to their high functional plasticity between M1 and M2 phenotypes, the evaluation of their response to decellularized ECM is mandatory. It is also necessary to analyze the immunocompetence of macrophages after contact with decellularized ECM materials to assess their functional role in a possible infection scenario. In this work, we studied the effect of four decellularized adipose matrices (DAMs) obtained from human and porcine AT by enzymatic or chemical methods on macrophage phenotypes and fungal phagocytosis. First, a thorough biochemical characterization of these biomaterials by quantification of remnant DNA, lipids, and proteins was performed, thus indicating the efficiency and reliability of both methods. The proteomic analysis evidenced that some proteins are differentially preserved depending on both the AT origin and the decellularization method employed. After exposure to the four DAMs, specific markers of M1 proinflammatory and M2 anti-inflammatory macrophages were analyzed. Porcine DAMs favor the M2 phenotype, independently of the decellularization method employed. Finally, a sensitive fungal phagocytosis assay allowed us to relate the macrophage phagocytosis capability with specific proteins differentially preserved in certain DAMs. The results obtained in this study highlight the close relationship between the ECM biochemical composition and the macrophage’s functional role.     

Design,Synthesis and Evaluation of 2,4-Diaminoquinazoline Derivatives as Potential Tubulin Polymerization Inhibitors

Microtubules are highly dynamic polymers composed of α- and β-tubulin proteins that have been shown to be potential therapeutic targets for the development of anticancer drugs. Currently, a wide variety of chemically diverse agents that bind to β-tubulin have been reported. Nocodazole (NZ) and colchicine (COL) are well-known tubulin-depolymerizing agents that have close binding sites in the β-tubulin. In this study, we designed and synthesized a set of nine 2,4-diaminoquinazoline derivatives that could occupy both NZ and COL binding sites. The synthesized compounds were evaluated for their antiproliferative activities against five cancer cell lines (PC-3, HCT-15, MCF-7, MDA-MB-231, and SK-LU-1), a noncancerous one (COS-7), and peripheral blood mononuclear cells (PBMC). The effect of compounds 4 e and 4 i on tubulin organization and polymerization was analyzed on the SK-LU-1 cell line by indirect immunofluorescence, western blotting, and tubulin polymerization assays. Our results demonstrated that both compounds exert their antiproliferative activity by inhibiting tubulin polymerization. Finally, a possible binding pose of 4 i in the NZ/COL binding site was determined by using molecular docking and molecular dynamics (MD) approaches. To our knowledge, this is the first report of non-N-substituted 2,4-diaminoquinazoline derivatives with the ability to inhibit tubulin polymerization.     

Anthocyanin profile of red fruits and black carrot juices,purees and concentrates by HPLC‐DAD‐ESI/MS‐QTOF

A fast and reliable method for anthocyanin extraction and identification by HPLC‐DAD‐ESI/MS‐QTOF was used to analyse the anthocyanin composition of commercial red fruit juices (blackberry, redcurrant and pomegranate), purees (strawberry, cherry and raspberry) and concentrates (elderberry, blueberry and red grape). The anthocyanin profile of black carrot juice is also reported. The extraction and analysis method allowed us to detect and quantify a wide range of individual anthocyanins in a simple and rapid way. Pelargonidin‐3‐glucoside was detected in redcurrant for the first time and petunidin‐3‐galactoside quantified for the first time in blueberries. Considering the health benefits that have been associated with anthocyanin consumption, all these fruit and vegetables processed products could appear as a good source of this group of phytochemical compounds for their direct consumption or their use as ingredients for the design of new food product or food supplements.     

Optimization of MRI Acquisition and Texture Analysis to Predict Physico-chemical Parameters of Loins by Data Mining

The main objective of this study was to configure the acquisition and analysis of low-field magnetic resonance imaging (MRI) to predict physico-chemical characteristics of Iberian loin, evaluating the use of different MRI sequences (spin echo, SE; gradient echo, GE; turbo 3D, T3D), computational texture feature methods (GLCM, NGLDM, GLRLM, GLCM + NGLDM + GLRLM), and data mining techniques (multiple linear regression, MLR; isotonic regression, IR). Moderate to very good correlation coefficients and low mean absolute error were found when applying MLR or IR on any method of computational texture features from MRI acquired with SE or GE. For T3D sequence, accurate results are only obtained by applying IR on GLCM or GLCM + NGLDM + GLRLM methods. Considering not only the accuracy of the methodology but also consumed time and required resources, the use of SE sequences for MRI acquisition, GLCM method for MRI texture analysis, and MLR could be indicated for prediction physico-chemical characteristics of loin.     

Quality assessment of commercial paprikas

Several quality parameters of smoked, oven‐dried and sun‐dried paprikas were studied. Smoked paprikas showed the highest American Spice Trade Association (ASTA) units and pigment concentrations, whereas oven‐dried paprikas showed the best browning index and red/yellow pigment ratio, and sun‐dried paprikas had the highest redness for reflected colour. The colour stability results showed drastic overall colour change (ΔE) and decreased redness and ASTA units of the sun‐dried and oven‐dried paprikas after two and 5 days of UV exposure. Degradation of the smoked paprika colour parameters was progressive, and they showed less degradation even after 30 days. The evaluation of the antioxidant activity showed higher DPPH radical‐scavenging capacity in the smoked samples. The profiles of volatile phenolic compounds of the smoked samples are responsible for the differences in the antioxidant capacity. The great differences in colour stability and antioxidant activity make smoked paprika the best choice for use in the food industry.     

Neuroprotective Role of Liver Growth Factor “LGF” in an Experimental Model of Cerebellar Ataxia

Cerebellar ataxias (CA) comprise a heterogeneous group of neurodegenerative diseases characterized by a lack of motor coordination. They are caused by disturbances in the cerebellum and its associated circuitries, so the major therapeutic goal is to correct cerebellar dysfunction. Neurotrophic factors enhance the survival and differentiation of selected types of neurons. Liver growth factor (LGF) is a hepatic mitogen that shows biological activity in neuroregenerative therapies. We investigate the potential therapeutic activity of LGF in the 3-acetylpiridine (3-AP) rat model of CA. This model of CA consists in the lesion of the inferior olive-induced by 3-AP (40 mg/kg). Ataxic rats were treated with 5 µg/rat LGF or vehicle during 3 weeks, analyzing: (a) motor coordination by using the rota-rod test; and (b) the immunohistochemical and biochemical evolution of several parameters related with the olivo-cerebellar function. Motor coordination improved in 3-AP-lesioned rats that received LGF treatment. LGF up-regulated NeuN and Bcl-2 protein levels in the brainstem, and increased calbindin expression and the number of neurons receiving calbindin-positive projections in the cerebellum. LGF also reduced extracellular glutamate and GABA concentrations and microglia activation in the cerebellum. In view of these results, we propose LGF as a potential therapeutic agent in cerebellar ataxias.     

Increasing espresso coffee brew antioxidant capacity using phenolic extract recovered from hazelnut skin waste

The simultaneous consumption of different classes of phytochemical antioxidants in the diet can result in more beneficial effects than when consumed alone. In the present study, the in vitro and in vivo antioxidative effects of espresso coffee brew (EC) (rich in chlorogenic acids) with added crude hazelnut skin phenolic extract (HSPE) from hazelnut skin waste (rich in flavonoids) were studied. Both post-brewing and pre-brewing phenolic-enriched espresso coffees (PE-ECs) were analysed for total phenols and screened for their in vitro antiradical ability. Moreover, the in vivo biological effect on the antioxidant potential of plasma in rats was evaluated. The PE-ECs showed increased both in vitro and in vivo antiradical activity proportional to the added HSPE. The in vivo experiments suggested that HSPE was much more antioxidant active than the phenolic fraction naturally contained in EC. Moreover, evidence of possible synergic effects of EC and HSPE phenolics was observed in vivo.     

Modeling Vehicle Miles Traveled on Local Roads Using Classification Roadway Spatial Structure

This paper models the relationship between vehicle miles traveled (VMT) on local and collector roads with an objective to predict local road VMT by using collector road VMT. Through a continuous approximation method typically used for vehicle routing, it first analytically reveals this relationship mainly as a function of roadway density ratios between multiple roadway classifications. This structural relationship suggests regression equations using density ratios or logarithmic values of them as the explanatory variables. The use of regression equations enables to account for varying spatial distributions of roadways and demand through parameter calibration. The proposed regression equations are proved good fits through computer simulation using distinct community road network topologies. In addition, practical data from Hennepin County of Minnesota, U.S.A. that encompasses Minneapolis indicates that our developed regression equations can work well.