Optimization of Diarylthiazole B‐Raf Inhibitors: Identification of a Compound Endowed with High Oral Antitumor Activity,Mitigated hERG Inhibition,and Low Paradoxical Effect

Aberrant activation of the mitogen‐activated protein kinase (MAPK)‐mediated pathway components, RAF‐MEK‐ERK, is frequently observed in human cancers and clearly contributes to oncogenesis. As part of a project aimed at finding inhibitors of B‐Raf, a key player in the MAPK cascade, we originally identified a thiazole derivative endowed with high potency and selectivity, optimal in vitro ADME properties, and good pharmacokinetic profiles in rodents, but that suffers from elevated hERG inhibitory activity. An optimization program was thus undertaken, focused mainly on the elaboration of the R¹ and R² groups of the scaffold. This effort ultimately led to N‐(4‐{2‐(1‐cyclopropylpiperidin‐4‐yl)‐4‐[3‐(2,5‐difluorobenzenesulfonylamino)‐2‐fluorophenyl]thiazol‐5‐yl}‐pyridin‐2‐yl)acetamide ( 20 ), which maintains favorable in vitro and in vivo properties, but lacks hERG liability. Besides exhibiting potent antiproliferative activity against only cell lines bearing B‐Raf V600E or V600D mutations, compound 20 also intriguingly shows a weaker “paradoxical” activation of MEK in non‐mutant B‐Raf cells than other known B‐Raf inhibitors. It also demonstrates very good efficacy in vivo against the A375 xenograft melanoma model (tumor volume inhibition >90 % at 10 mg kg^?1); it is therefore a suitable candidate for preclinical development.     

Acellular Dermal Matrix Used in Diabetic Foot Ulcers: Clinical Outcomes Supported by Biochemical and Histological Analyses

Diabetic foot ulcer (DFU) is a diabetes complication which greatly impacts the patient’s quality of life, often leading to amputation of the affected limb unless there is a timely and adequate management of the patient. DFUs have a high economic impact for the national health system. Data have indeed shown that DFUs are a major cause of hospitalization for patients with diabetes. Based on that, DFUs represent a very important challenge for the national health system. Especially in developed countries diabetic patients are increasing at a very high rate and as expected, also the incidence of DFUs is increasing due to longevity of diabetic patients in the western population. Herein, the surgical approach focused on the targeted use of the acellular dermal matrix has been integrated with biochemical and morphological/histological analyses to obtain evidence-based information on the mechanisms underlying tissue regeneration. In this research report, the clinical results indicated decreased postoperative wound infection levels and a short healing time, with a sound regeneration of tissues. Here we demonstrate that the key biomarkers of wound healing process are activated at gene expression level and also synthesis of collagen I, collagen III and elastin is prompted and modulated within the 28-day period of observation. These analyses were run on five patients treated with Integra^® sheet and five treated with the injectable matrix Integra^® Flowable, for cavitary lesions. In fact, clinical evaluation of improved healing was, for the first time, supported by biochemical and histological analyses. For these reasons, the present work opens a new scenario in DFUs treatment and follow-up, laying the foundation for a tailored protocol towards complete healing in severe pathological conditions.     

Solubilization and Activity Detection in Polyacrylamide Gels of a Membrane‐Bound Esterase from an Oenological Strain of Saccharomyces cerevisiae

Due to the importance of esters in determining wine aroma, the presence of different esterases in soluble and insoluble cell fractions of an oenological strain of Saccharomyces cerevisiae was studied. Cells of an oenological yeast strain were separated into three fractions corresponding to the cytoplasm, periplasm and plasma membrane, all showing esterase activity. The conditions for optimal solubilization of the membrane‐bound esterase were found, as well as those allowing its electrophoretic analysis, using the detergent disodium‐n‐lauryl‐β‐iminodipropionate (Deriphat) in the cathodic buffer. Comparison of the activity bands detected on gels revealed the presence of different esterase iso‐forms in the three sub‐cellular fractions. The method can also be used as the first electrophoretic step in two‐dimensional PAGE. Therefore, the procedures described are also a promising tool for the study of the yeast enzymes in relation to their effects during winemaking.     

Effect of growth factors and lactogenic hormones on expression of plasminogen activator-related genes and cell proliferation in a bovine mammary epithelial cell line

There is conflicting evidence in the literature as to whether up-regulation of urokinase plasminogen activator (u-PA) expression is related to bovine mammary epithelial cell growth. The role of u-PA receptor (u-PAR) and that of the plasminogen activator inhibitors type 1 and type 2 (PAI-1 and PAI-2) in bovine mammary epithelial cell proliferation is not known. The effect of growth factors and various hormones known to affect mammary function on expression of u-PA, u-PAR, PAI-1, PAI-2 and cell proliferation using the BME-UV1 bovine mammary epithelial cell line was examined. Cell proliferation was measured using the MTT assay and direct cell enumeration. Results showed that both IGF-1 and EGF increased cell proliferation but EGF was a more potent mitogen than IGF-1. Furthermore, IGF-1 increased by 2-fold expression of both u-PA and u-PAR while EGF increased by 3·8-fold the expression of only u-PAR. Both growth factors had no effect on expression of PAI-1 and PAI-2. In a manner consistent with changes in gene expression, EGF and to a lesser extent IGF-1 up-regulated total cell associated, membrane-bound and secreted u-PA activity. Thus, a strong correlation exists between u-PAR gene expression along with the activity of u-PA present on cell membranes and cell proliferation. Dexamethasone, prolactin and surprisingly insulin had no effect on cell proliferation. Dexamethasone alone and when combined with insulin or prolactin up-regulated gene expression of both PAI- and PAI-2 but not that of u-PA and u-PAR. Decreased total cell-associated, membrane-bound and secreted u-PA activity was detected in cells cultured in the presence of dexamethasone when combined with insulin or prolactin. However no such effect was observed in the presence of dexamethasone alone. Thus, dexamethasone acting synergistically with prolactin or insulin inhibits the activation of the plasmin-plasminogen system but this inhibition is not correlated with any changes in cell proliferation.     

Vacuum-packed ripened sausages: evolution of volatile compounds during storage

BACKGROUND: Various studies have shown the ability of volatile compounds to influence the quality of charcuterie products because of their effect on sensorial properties. To the best of our knowledge, there are no data in the scientific literature concerning the evolution of volatile compounds of vacuum‐packed ripened sausages over time. The aim of this work was to evaluate the evolution of volatile compounds in vacuum‐packed ripened sausages over time, considering that this kind of packaging is the most commonly used to prolong the shelf‐life of these products. RESULTS: Sixty‐nine volatile compounds were identified. Those derived from spices represented the prevailing compounds at the beginning of storage and significantly decreased (P < 0.001) during 5 months of storage, as a result of the well‐known antioxidant activity. Conversely, compounds derived from the autoxidation of lipids, which are among those chiefly responsible of sensorial properties of meat products, significantly increased (P < 0.001). CONCLUSION: The research demonstrated that vacuum packaging influenced the evolution of volatile compounds during storage of ripened sausages. In particular, the increase of volatile compounds derived from autoxidation of the lipid fraction indicated that oxidative processes involved the fatty fraction of ripened sausages also in the scarcity of oxygen. Copyright © 2011 Society of Chemical Industry     

Characterisation of Photocathodes Based on Pb Thin Film Deposited by UV Pulsed Laser Ablation简

This work deals with the deposition of lead(Pb) thin films by the U V pulsed laser ablation technique,for their further use as photocathode devices in superconducting radio frequency guns.Scanning electron microscopy and atomic force microscopy analyses were performed to study the morphological features of Pb thin films deposited on Si(100) and Nb substrates.The films showed a granular structure with a nearly fully covered surface only for that one deposited on Nb substrata X-ray diffraction measurements indicate the growth of polycrystalline Pb thin films with a preferential orientation along(111) planes.Results of the photoemission performance of Pb thin film deposited on Nb substrate showed a very encouraging average value of quantum efficiency of 6×10~(-5) through a single-photon absorption process,promoting further studies in the realisation of Pb photocathodes by this techniqua     

Assessment of the Influence of the Decanter Set-up During Continuous Processing of Olives at Different Pigmentation Index

New generation decanters allow operators to make real-time adjustments during the virgin olive oil extraction process in order to gain the best performance possible. However, the opportunity to act in line requires a deep understanding of the consequences of changing machine parameters. To this purpose, an experiment was carried out at industrial scale. The decanter feed rate (Fr), ranging from 4075 to 5820 kg h^?1, the bowl/screw conveyor differential speed (?n), set at 18 and 22, and two ripening degrees of the olives were considered as process variables. Two combinations Fr-?n, namely 4620 kg h^?1 at ?n-18 and 5210 kg h^?1 at ?n-22, were found to similarly maximize the process efficiency, regardless the raw material features. After pointing out the best working settings, the corresponding virgin olive oils were compared. The analysis of variance showed that peroxide value, K ₂₃₂, K ₂₇₀, phenols, chlorophylls, β-tocopherol, fruity and bitter notes, and C6 volatile compounds were significantly affected by the machine parameters. An inverse proportionality was observed between the combination Fr-?n and the phenolic compounds. On the whole, the sampling factor exerted a larger influence on the product quality than the decanter set-up.     

Influence of the exposure to light on extra virgin olive oil quality during storage

An experimental investigation was carried out to study the influence that exposure to light has on the quality of extra virgin oil during a 12-month storage period by comparing it with the quality of extra virgin oil stored in the dark. The results showed that the oils stored in the light had significantly lower tocopherol, carotenoid and chlorophyll contents than did the same oils kept in the dark. Moreover, the oils stored in the dark mainly contained primary oxidation products, while the oils kept in the light contained secondary oxidation products as confirmed by the K₂₇₀ values which exceeded the legal limits even after purification by means of alumina. Overall, the results obtained showed that the shelf life of the oils exposed to light is shorter than that of oils kept in the dark, and that after only 2 months of exposure to light the oils examined could no longer be considered as extra virgin.     

Photoresponsive Adhesives Based on Arylazoisoxazoles-Containing Polymers

Adhesives exist in many forms and are essential in the daily life, in industrial processes, and in healthcare, calling for continuous improvements in terms of application fields, strength, biocompatibility, recyclability, and reversibility. Regarding the last two points, embedding photoresponsive molecules into the adhesive allows the use of light to attach or detach reversibly different surfaces. Photoresponsive adhesives can be removed without the use of high temperature or solvents, which is ideal for delicate environments. To date, azobenzenes are the most explored chromophore to prepare smart adhesive formulations. Here, the synthesis of polymer containing arylazoisoxazoles (AIZs) — azobenzene analogs containing a heterocycle — and their application as adhesives is described. Mechanical tests demonstrate an enhancement of the adhesion strength from 0.39 to 2.79 MPa, passing from low molecular weight monomer to selected AIZs acrylate polymer. Irradiation with UV induces the formation of the cis isomer with a consequent drop of more than 70% in the adhesion strength. The different compounds result as possible candidates as adhesives on the basis of the increased load that can be held, together with the possibility to obtain joints that can be either stable or detachable under light irradiation.