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The galvanic deposition method was used to deposit Hydroxyapatite/Chitosan/Collagen coatings on 304 stainless steel. Galvanic deposition is an alternative and valid way to fabricate bio-coatings with high biocompatibility and good anticorrosion properties. Physical-chemical characterizations were carried out to investigate chemical composition and morphology of the samples. Coatings consist of a mixture of calcium phosphate (Brushite and Hydroxyapatite) with chitosan and collagen. Corrosion tests were performed in the simulated body fluid (SBF) after different aging times. Results show that, in comparison with bare 304 stainless steel, coating shifts corrosion potential to anodic values and reduces corrosion current density. Nevertheless, the aging in SBF led to a completely conversion of brushite into hydroxyapatite. The release of metal ions, measured after 21 days of aging in SBF solution, is very low due to the presence of coating that slow-down the corrosion rate of steel.

During the last decades, biomaterials have been deeply studied to fabricate and improve coatings for biomedical devices. Metallic materials, especially in the orthopedic field, represent the most common materials used for different type of devices thanks to their good mechanical properties. Nevertheless, low/medium resistance to corrosion and low osteointegration ability characterizes these materials. To overcome these problems, the use of biocoatings on metals substrate is largely diffused. In fact, biocoatings have a key role to confer biocompatibility features, to inhibit corrosion and thus improve the lifetime of implanted devices. In this work, the attention was focused on Hydroxyapatite-Chitosan (HA/CS) and Hydroxyapatite-Polyvinylacetate (HA/PVAc) composites, that have been studied as biocoatings for 304 SS based devices. Hydroxyapatite was selected for its osteoconductivity due to its chemical structure similar to bones. Furthermore, Chitosan and Polyvinylacetate are largely used yet in medical field (e.g. antibacterial agent or drug deliver) and in this work were used to create a synergic interaction with hydroxyapatite to increase the strength and bioactivity of coating. Despite bio-coatings were obtained by different techniques, in this work, they were fabricated by galvanic deposition process that has different advantages, among which it does not require external power supply. It is a spontaneous electrochemical reaction in which materials with different standard electrochemical potential were short-circuited and immersed in an electrolytic solution. Electrons supplied by the anodic reaction at the less noble electrode flow to cathode where they oxidize the less noblest ions in solution. SEM, EDS, XRD and RAMAN were performed for chemical-physics characterization of biocoatings. Polarization and impedance measurements have been also carried out to evaluate corrosion behavior. Besides, in-vitro cytotoxicity assays have been done for the biological features.

© 2015 Elsevier Inc.Protein kinase C (PKC) isozymes have remained elusive cancer targets despite the unambiguous tumor promoting function of their potent ligands, phorbol esters, and the prevalence of their mutations. We analyzed 8% of PKC mutations identified in human cancers and found that, surprisingly, most were loss of function and none were activating. Loss-of-function mutations occurred in all PKC subgroups and impeded second-messenger binding, phosphorylation, or catalysis. Correction of a loss-of-function PKCβ mutation by CRISPR-mediated genome editing in a patient-derived colon cancer cell line suppressed anchorage-independent growth and reduced tumor growth in a xenograft model. Hemizygous deletion promoted anchorage-independent growth, revealing that PKCβ is haploinsufficient for tumor suppression. Several mutations were dominant negative, suppressing global PKC signaling output, and bioinformatic analysis suggested that PKC mutations cooperate with co-occurring mutations in cancer drivers. These data establish that PKC isozymes generally function as tumor suppressors, indicating that therapies should focus on restoring, not inhibiting, PKC activity.

To define novel pathways that regulate susceptibility to tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) in non-small cell lung cancer (NSCLC), we have performed genome-wide expression profiling of microRNAs (miRs). We show that in TRAIL-resistant NSCLC cells, levels of different miRs are increased, and in particular, miR-221 and -222. We demonstrate that these miRs impair TRAIL-dependent apoptosis by inhibiting the expression of key functional proteins. Indeed, transfection with anti-miR-221 and -222 rendered CALU-1-resistant cells sensitive to TRAIL. Conversely, H460-sensitive cells treated with -221 and -222 pre-miRs become resistant to TRAIL. miR-221 and -222 target the 3'-UTR of Kit and p27(kip1) mRNAs, but interfere with TRAIL signaling mainly through p27(kip1). In conclusion, we show that high expression levels of miR-221 and -222 are needed to maintain the TRAIL-resistant phenotype, thus making these miRs as promising therapeutic targets or diagnostic tool for TRAIL resistance in NSCLC.

Akt is a serine threonine kinase with a major role in transducing survival signals and regulating proteins involved in apoptosis. To find new interactors of Akt involved in cell survival, we performed a two-hybrid screening in yeast using human full-length Akt c-DNA as bait and a murine c-DNA library as prey. Among the 80 clones obtained, two were identified as Bcl-w. Bcl-w is a member of the Bcl-2 family that is essential for the regulation of cellular survival, and that is up-regulated in different human tumors, such as gastric and colorectal carcinomas. Direct interaction of Bcl-w with Akt was confirmed by immunoprecipitation assays. Subsequently, we addressed the function of this interaction: by interfering with the activity or amount of Akt, we have demonstrated that Akt modulates the amount of Bcl-w protein. We have found that inhibition of Akt activity may promote apoptosis through the downregulation of Bcl-w protein and the consequential reduction in interaction of Bcl-w with proapoptotic members of the Bcl-2 family. Our data provide evidence that Bcl-w is a new member of the Akt pathway and that Akt may induce anti-apoptotic signals at least in part through the regulation of the amount and activity of Bcl-w.

Aims The combined prophylactic strategy of sodium bicarbonate plus N-acetylsyteine (NAC) seems to be effective in preventing contrast induced acute kidney injury (CI-AKI) in patients at low-to-medium risk. However, in patients at high and very high risk the rate of CI-AKI is still high. In this subset of patients the anticipated advantages of the RenalGuard(tm) System should be investigated. The RenalGuard(tm) System (PLC Medical Systems, Inc., Franklin, MA, USA) is a real-time measurement and real time matched fluid replacement device designed to accommodate the RenalGuard therapy, which is based on the theory that creating and maintaining a high urine output is beneficial by allowing a quick elimination of contrast media, and, therefore, reducing its toxic effects. Methods and results The REMEDIAL II trial is a randomised, multicentre, investigator-sponsored trial addressing the hypothesis that the RenalGuard System is superior to the prophylaxis with sodium bicarbonate infusion plus NAC in preventing CI-AKI in high and very high risk patients. Consecutive patients with chronic kidney disease (CKD) and at high to very high risk for CI-AKI, referred to our institutions for coronary and/or peripheral procedures, will be randomly assigned to 1) prophylactic administration of sodium bicarbonate plus NAC (control group) and 2) RenalGuard System treatment (RenalGuard group). All enrolled patients must have an estimated glomerular filtration rate ≤ 30 ml/min/1.73 m2 and/or a contrast nephropathy risk score ≥ 11. In all cases iodixanol (an iso-osmolar, non-ionic contrast agent) will be administered. The primary endpoint is an increase of ≥ 0.3 mg/dL in the serum creatinine concentration 48 hours after the procedure. Conclusions The REMEDIAL II trial will give important answers on how to prevent CI-AKI in high and very high risk patients undergoing contrast media exposure.

In spite of growing evidence linking vitamin D(3) levels to mental health disorders, little is known about its direct targets in the brain. This study set out to investigate targets of vitamin D(3) in a human brain stem cell line. We employed arrays with antibodies directed against more than 600 structural and signalling proteins, including phospho-variants. Over 180 proteins responded to vitamin D(3), such as cyclin-dependent protein-serine kinase 1/2, epidermal growth factor receptor-tyrosine kinase, protein kinase A, protein-serine kinase Bgamma and protein-serine kinase Calpha. PEA-15 (phosphoprotein enriched in astrocytes-15 kD, also known as PED), known to be involved in various anti-proliferative and anti-apoptotic effects, was strongly up-regulated. In silico promoter analysis revealed conserved binding sites for vitamin D(3) receptor, suggesting a strong vitamin D(3) dependency of the PEA-15 promoter. PEA-15 up-regulation by vitamin D(3) could be confirmed by Western blot in two different cell lines. Analysis of mRNA and protein phosphorylation status of PEA-15 suggests that increased PEA-15 promoter activity and increased protein stabilization contribute to the overall rise of PEA-15 protein. In a functional test of this novel pathway, we demonstrated that vitamin D(3) was able to rescue cells from TRAIL-induced apoptosis through regulation of the PEA-15 expression and function. Summarized, our study presents novel targets of vitamin D(3) relevant for apoptosis and cell proliferation, and thus strongly supports a function of vitamin D(3) in the brain that impacts on processes highly relevant for major neurological disorders.

The performance of electrochemical sensors using an in situ pH control technique for detection of mercury and copper in neutral solutions is described herein. Sensors are comprised of two distinct parallel gold interdigitated microband electrodes each of which may be polarised separately. Biasing one interdigitated “protonator” electrode sufficiently positive to begin water electrolysis, resulted in the production of H+ ions, which, consequently droped the interfacial pH at the other second interdigitated “sensing” electrode. This decrease in pH permitted the electrodeposition (and consequent stripping) of metals at a sensing electrode without the need to acidify the whole test solution. In this work, the local pH could be adjusted in the acidic pH range in a stable and reproducible way just by tailoring the polarization of the protonator electrode. Using this approach, a linear range for copper 5 to 100 ppb and for mercury 1 to 75 ppb were obtained. The sensors also have an extremely high sensitivity for the metals. The in-situ pH control, coupled with electrochemical stripping, allowed metal detection in a complex water matrix, e.g., river water without the need for sample pre-treatment. The electrochemical results were confirmed by comparison to those obtained using inductively coupled plasma – optical emission spectroscopy. A very good agreement was observed between both sets of results. The electrode reproducibility was high (RSD < 10%) and the metals could be co-detected simultaneously. Thus, this work shows a fast and easy approach for in-situ pH control for multi metal detection using solid state sensors.

Nanostructured Pb electrodes consisting of nanowire arrays were obtained by electrodeposition, to be used as negative electrodes for lead–acid batteries. Reduced graphene oxide was added to improve their performances. This was achieved via the electrochemical reduction of graphene oxide directly on the surface of nanowire arrays. The electrodes with and without reduced graphene oxide were tested in a 5 M sulfuric acid solution using a commercial pasted positive plate and an absorbed glass mat separator in a zero-gap configuration. The electrodes were tested in deep cycling conditions with a very low cut-off potential. Charge–discharge tests were performed at 5C. The electrode with reduced graphene oxide outperformed the electrode without reduced graphene oxide, as it was able to work with a very high utilization of active mass and efficiency. A specific capacity of 258 mAhg−1–very close to the theoretical one–was achieved, and the electrode lasted for more than 1000 cycles. On the other hand, the electrode without reduced graphene oxide achieved a capacity close to 230 mAhg−1, which corresponds to a 90% of utilization of active mass.

In this work, composite coatings of chitosan and silver nanoparticles were presented as an antibacterial coating for orthopedic implants. Coatings were deposited on AISI 304L using the galvanic deposition method. In galvanic deposition, the difference of the electrochemical redox potential between two metals (the substrate and a sacrificial anode) has the pivotal role in the process. In the coupling of these two metals a spontaneous redox reaction occurs and thus no external power supply is necessary. Using this process, a uniform deposition on the exposed area and a good adherence of the composite coating on the metallic substrate were achieved. Physical-chemical characterizations were carried out to evaluate morphology, chemical composition, and the presence of silver nanoparticles. These characterizations have shown the deposition of coatings with homogenous and porous surface structures with silver nanoparticles incorporated and distributed into the polymeric matrix. Corrosion tests were also carried out in a simulated body fluid at 37 °C in order to simulate the same physiological conditions. Corrosion potential and corrosion current density were obtained from the polarization curves by Tafel extrapolation. The results show an improvement in protection against corrosion phenomena compared to bare AISI 304L. Furthermore, the ability of the coating to release the Ag+ was evaluated in the simulated body fluid at 37 °C and it was found that the release mechanism switches from anomalous to diffusion controlled after 3 h.

Organometallic perovskites are one of the most investigated materials for high-efficiency thin-film devices to convert solar energy and supply energy. In particular, methylammonium lead iodide has been used to realize thin-film perovskite solar cells, achieving an efficiency higher than 20%. Different fabrication procedures based on the spin-coating technique have been proposed, which do not ensure homogenous morphologies. In this work, we present a scalable process to fabricate methylammonium lead iodide thin films directly on conductive substrates, consisting of electrodeposition and two subsequent chemical conversions. A thorough investigation of the morphological, structural and compositional properties of the layer is performed after each fabrication step. It is demonstrated that this method allows fine control of the thickness of the layer by tuning the cell parameters during the electrodeposition step. X-ray diffraction patterns and energy-dispersive X-ray analysis indicate the achievement of high-purity methylammonium lead iodide layers. Micro-Raman analyses were used to demonstrate the formation of methylammonium lead iodide. Finally, ultraviolet-visible absorption spectra were acquired to determine the optical band edge of the layer ( 1.56 eV) and the absorbance of methylammonium lead iodide as a function of the film thickness. As expected, the material exploits excellent optical properties, achieving an absorption ≥ 99.9% in the entire visible range for a layer thickness of 1.3 µm. The results presented here pave the way for the application of cost-friendly solution-based processes to fabricate high-quality perovskite solar cells.

In this work, the galvanic deposition method is used to deposit coatings of brushite/hydroxyapatite/polyvinyl acetate on 304 stainless steel. Coatings are obtained at different temperatures and with different sacrificial anodes, consisting of a mixture of brushite and hydroxyapatite. Samples are aged in a simulated body fluid (SBF), where a complete conversion of brushite into hydroxyapatite with a simultaneous change in morphology and wettability occurred. The corrosion tests show that, compared with bare 304, the coating shifts Ecorr to anodic values and reduces icorr Ecorr, and icorr has different values at different aging times due to chemical interactions at the solid/liquid interface. The best performing deposits are those obtained by using Al as the sacrificial anode. The metal ion release, measured after 21 days of aging, is very low and is attributable to the presence of a coating that slows the steel corrosion. Coating cytotoxicity is investigated through cell viability assays with MC3T3-E1 osteoblastic cells. The results reveal a high cytocompatibility comparable to that of a pure cell culture medium.

AIMS: Contrast media (CM) induce a direct toxic effect on renal tubular cells. This toxic effect may have a role in the pathophysiology of contrast nephropathy. METHODS AND RESULTS: We evaluated (i) the cytotoxicity of CM [both low-osmolality (LOCM) and iso-osmolality (IOCM)], of iodine alone, and of an hyperosmolar solution (mannitol 8%) on human embryonic kidney (HEK 293), porcine proximal renal tubular (LLC-PK1), and canine Madin-Darby distal tubular renal (MDCK) cells; and (ii) the effectiveness of various antioxidant compounds [n-acetylcysteine (NAC), ascorbic acid and sodium bicarbonate] in preventing CM cytotoxicity. The cytotoxicity of CM was assessed at different time points, with different methods: cell viability, DNA laddering, flow cytometry, and caspase activation. Both LOCM and IOCM produced a concentration- and time-dependent increase in cell death as assessed by the different methods. On the contrary, iodine alone and hyperosmolar solution did not induce any significant cytotoxic effect. There was not any significant difference in the cytotoxic effect between LOCM and IOCM. Furthermore, both LOCM and IOCM caused a marked increase in caspase-3 and -9 activities and poly(ADP-ribose) fragmentation, while no effect on caspase-8/-10 was observed, thus indicating that the CM activated apoptosis mainly through the intrinsic pathway. Both CM induced an increase in protein expression levels of pro-apoptotic members of the Bcl2 family (Bim and Bad). NAC and ascorbic acid but not sodium bicarbonate had a dose-dependent protective effect on renal cells after 3 h incubation with high dose (200 mg iodine/mL) of both LOCM and IOCM. CONCLUSION: Both LOCM and IOCM induce a dose-dependent renal cell apoptosis. NAC and ascorbic acid but not sodium bicarbonate prevent this contrast-induced apoptosis.

PED (phosphoprotein enriched in diabetes) is a death-effector domain (DED) family member with a broad anti-apoptotic action. PED inhibits the assembly of the death-inducing signalling complex (DISC) of death receptors following stimulation. Recently, we reported that the expression of PED is increased in breast cancer cells and determines the refractoriness of these cells to anticancer therapy. In the present study, we focused on the role of PED in non-small cell lung cancer (NSCLC), a tumour frequently characterized by evasion of apoptosis and drug resistance. Immunohistochemical analysis of a tissue microarray, containing 160 lung cancer samples, indicated that PED was strongly expressed in different lung tumour types. Western blotting performed with specimens from NSCLC-affected patients showed that PED was strongly up-regulated (>6 fold) in the areas of tumour compared to adjacent normal tissue. Furthermore, PED expression levels in NSCLC cell lines correlated with their resistance to tumour necrosis factor related apoptosis-inducing ligand (TRAIL)-induced cell death. The involvement of PED in the refractoriness to TRAIL-induced cell death was investigated by silencing PED expression in TRAIL-resistant NSCLC cells with small interfering (si) RNAs: transfection with PED siRNA, but not with cFLIP siRNA, sensitized cells to TRAIL-induced cell death. In conclusion, PED is specifically overexpressed in lung tumour tissue and contributes to TRAIL resistance.

Killing of tumor cells by cytotoxic therapies, such as chemotherapy or gamma-irradiation, is predominantly mediated by the activation of apoptotic pathways. Refractoriness to anticancer therapy is often due to a failure in the apoptotic pathway. The mechanisms that control the balance between survival and cell death in cancer cells are still largely unknown. Tumor cells have been shown to evade death signals through an increase in the expression of antiapoptotic molecules or loss of proapoptotic factors. We aimed to study the involvement of PED, a molecule with a broad antiapoptotic action, in human breast cancer cell resistance to chemotherapeutic drugs–induced cell death. We show that human breast cancer cells express high levels of PED and that AKT activity regulates PED protein levels. Interestingly, exogenous expression of a dominant-negative AKT cDNA or of PED antisense in human breast cancer cells induced a significant down-regulation of PED and sensitized cells to chemotherapy-induced cell death. Thus, AKT-dependent increase of PED expression levels represents a key molecular mechanism for chemoresistance in breast cancer.

B-cell chronic lymphocytic leukaemia (B-CLL) cells fail to undergo apoptosis. The mechanism underlying this resistance to cell death is still largely unknown. Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) effectively kills tumour cells but not normal cells, and thus represents an attractive tool for the treatment of cancer. Unfortunately, lymphocytes from B-CLL patients are resistant to TRAIL-mediated apoptosis. Thus, we aimed to study the involvement of PED, a DED-family member with a broad antiapoptotic action, in this resistance. We demonstrate that B lymphocytes obtained from patients with B-CLL express high levels of PED. Treatment of B-CLL cells with specific PED antisense oligonucleotides, a protein synthesis inhibitor or HDAC inhibitors, induced a significant downregulation of PED and sensitized these cells to TRAIL-induced cell death. These findings suggest a direct involvement of PED in resistance to TRAIL-induced apoptosis in B-CLL. It also identifies this DED-family member as a potential therapeutic target for this form of leukaemia.