Tribocorrosion Properties of PEO Coatings Produced on AZ91 Magnesium Alloy with Silicate- or Phosphate-Based Electrolytes

In this work, the tribocorrosion behavior of plasma electrolytic oxidation (PEO)-coated AZ91 samples was studied. In particular, two different coatings were produced and compared. One was obtained with an alkaline electrolyte containing sodium phosphate, whereas the other one was produced with an alkaline electrolyte containing sodium silicate. The coatings were characterized with SEM-EDS and XRD techniques, and after the tribocorrosion tests, the wear scars were analyzed with SEM-EDS. The tribocorrosion behavior was evaluated measuring the OCP during a pin on disk test performed in an aggressive environment. Moreover, potentiodynamic polarization and electrochemical impedance spectroscopy tests were performed, to evaluate the corrosion resistance of the different samples in the absence of wear phenomena. The behavior of all the PEO-treated specimens was compared with the one of the untreated sample. A remarkable increase in the tribocorrosion performances after the PEO treatments was observed. Moreover, the samples obtained with the electrolyte containing silicates showed higher tribocorrosion performances

Validation of a Novel Three-Dimensional (3D Fusion) Gross Sampling Protocol for Clear Cell Renal Cell Carcinoma to Overcome Intratumoral Heterogeneity: The Meet-Uro 18 Study

We aimed to overcome intratumoral heterogeneity in clear cell renal cell carcinoma (clearRCC). One hundred cases of clearRCC were sampled. First, usual standard sampling was applied (1 block/cm of tumor); second, the whole tumor was sampled, and 0.6 mm cores were taken from each block to construct a tissue microarray; third, the residual tissue, mapped by taking pieces 0.5 × 0.5 cm, reconstructed the entire tumor mass. Precisely, six randomly derived pieces of tissues were placed in each cassette, with the number of cassettes being based on the diameter of the tumor (called multisite 3D fusion). Angiogenic and immune markers were tested. Routine 5231 tissue blocks were obtained. Multisite 3D fusion sections showed pattern A, homogeneous high vascular density (10%), pattern B, homogeneous low vascular density (8%) and pattern C, heterogeneous angiogenic signatures (82%). PD-L1 expression was seen as diffuse (7%), low (33%) and absent (60%). Tumor-infiltrating CD8 scored high in 25% (pattern hot), low in 65% (pattern weak) and zero in 10% of cases (pattern desert). Grading was upgraded in 26% of cases (G3-G4), necrosis and sarcomatoid/rhabdoid characters were observed in, respectively, 11 and 7% of cases after 3D fusion (p = 0.03). CD8 and PD-L1 immune expressions were higher in the undifferentiated G4/rhabdoid/sarcomatoid clearRCC subtypes (p = 0.03). Again, 22% of cases were set to intermediate to high risk of clinical recurrence due to new morphological findings of all aggressive G4, sarcomatoid/rhabdoid features by using 3D fusion compared to standard methods (p = 0.04). In conclusion, we propose an easy-to-apply multisite 3D fusion sampling that negates bias due to tumor heterogeneity

Overdentures on implants placed in bone augmented with fresh frozen bone

Introduction In the last decade several studies have been performed to evaluate the clinical outcome of one or two stage loaded implants supporting overdentures. Aim Since fresh frozen bone (FFB) has an ever-increasing number of clinical applications and few reports are available on implants inserted into FFB, we performed a retrospective study on fixtures inserted in FFB and bearing overdentures. Methods In the period between December 2003 and December 2006, 17 patients (14 females and 3 males with a median age of about 56 years) were grafted and 60 implants inserted thereafter. A total of 17 overdentures were delivered: 8 in the mandible and 9 in the maxilla. Multiple implant systems were used: 22 Double etched, 7 SLA, 9 Anodic oxidized, and 22 CaPo4 ceramic-blasted. Implant diameter ranged from 3.25 to 4.3 mm and length from 11.5 to 16.0 mm. Implants were inserted to replace 23 incisors, 9 cuspids, 20 premolars and 8 molars. Results No implants were lost (i.e. survival rate = 100%) and no differences were detected among the studied variables. Kaplan Meier algorithm and Cox regression did not reveal any statistical differences among the studied variables also as regards the success rate. Conclusion Implants inserted FFB and bearing overdentures have a high survival rate and success rates, which are comparable to those of implants inserted in non-grafted bone. FFB bone is a reliable material for alveolar ridge augmentation. No difference was detected among removable prostheses supported by 2 or more implants

Microstructural and Corrosion Properties of PEO Coated Zinc-Aluminized (ZA) Steel

Plasma Electrolytic Oxidation (PEO) is a surface treatment, similar to anodizing, that produces thick oxide films on the surface of metals. In the present work, PEO coatings were obtained on zinc-aluminized (ZA) carbon steel using a solution containing sodium silicate and potassium hydroxide as electrolyte, and working with high current densities and short treatment times in Direct Current (DC) mode. The thickness of the coating, as well as the surface morphology, were strongly influenced by the process parameters, with different dissolution grades of the ZA layer depending on the current density and treatment time. A compromise between thickness and porosity of the coating was found with low current density/long treatment time or high current density/short treatment time. The PEO layer was mainly composed of aluminum oxides and silicon compounds. The corrosion resistance increased remarkably in the samples with the PEO coating. These PEO coated samples are suitable for sealing treatments that further increase their corrosion properties or will be also an ideal substrate for commercial painting, assuring improved mechanical adhesion and protection even in the presence of damages

RASSF1 tumor suppressor gene in pancreatic ductal adenocarcinoma: correlation of expression, chromosomal status and epigenetic changes

Background: The Ras Association Domain Family Member 1 (RASSF1) is one of the most frequently reported methylation-inactivated tumor suppressor genes in primary pancreatic ductal adenocarcinomas (PDAC). Limited information is still available about the impact of RASSF1 gene silencing on the expression of its different isoforms in neoplastic cells. Methods: A series of 96 primary PDAC, with known clinico-pathological parameters, was tested for RASSF1 methylation status by methylation-specific PCR, RASSF1 locus copy number alterations by fluorescence in situ hybridization, and Rassf1a protein expression by immunohistochemistry. A further series of 14 xenografted primary PDAC and 8 PDAC-derived cell lines were tested to obtain a detailed methylation mapping of CpG islands A and C of the RASSF1 locus by pyrosequencing and to evaluate the expression of Rassf1 variants by qRT-PCR. Results: Methylation of CpG island A of the RASSF1 gene was observed in 35% of the tumors and allelic loss of RASSF1 locus was seen in 30 disomic and in 20 polysomic cases (52%). Rassf1a immunohistochemical expression was downregulated in half of primary PDAC, and this downregulation was neither correlated with methylation of RASSF1 promoter nor with RASSF1 copy number alterations. RASSF1 status did not influence patients' prognosis. The expression of the seven RASSF1 isoforms in xenografts and cell lines showed that RASSF1A, RASSF1B, and RASSF1C isoforms were present in all xenografts and cell lines, whereas RASSF1D, RASSF1E, and RASSF1F isoforms were variably expressed among samples. RASSF1G was never expressed in either xenografts or cell lines. The variable expression of RASSF1 isoforms in PDAC xenografts and cell lines was not dependent on RASSF1 methylation status of CpG islands A and C. Conclusions:RASSF1 alterations occurring in PDAC mainly consist in variations of expression of the different isoforms. Different genetic mechanisms seem to contribute to RASSF1 deregulation in this setting, but RASSF1 methylation does not seem to substantially affect RASSF1 isoforms expression

Is the Rivermead Mobility Index a suitable outcome measure in lower limb amputees?--A psychometric validation study.

Objective: To examine the internal consistency, validity, responsiveness and test scalability of the Rivermead Mobility Index. Design: Methodological research (consecutive sampling, prospective longitudinal study). Patients: 140 unilateral lower limb amputees (79 above-knee and 61 below-knee). Methods: The Rivermead Mobility Index was administered to all patients at the beginning (T0) and at the end (T2) of the prosthetic training. In 70 of the patients, the Functional Independence Measure and a timed walking test were also carried out. Results: The Cronbach's alpha of the Rivermead Mobility Index was 0.85 and the item-to-total correlation coefficients rpb ranged from 0.33 to 0.74 (p 0.0001), for the items considered, at T0; 4 correlations were not calculated due to the extremely low variability of some item responses (mode 98%). The correlation (rs) of Rivermead Mobility Index score with the motor subscale of the Functional Independence Measure was 0.83 at T0 and 0.69 at T2 (p 0.0001, for both) and that with timed walking test 0.70 (p 0.0001) at T2. The effect size was 1.35. The scalability coefficients were below the limits of acceptability. Conclusion: When applied in lower limb amputees, the Rivermead Mobility Index is an ordinal measure with adequate levels of a series of psychometric properties, which seems more useful for epidemiological studies than for clinical decision-making in single patients. Further steps should be considered to improve its item selection, response format and scaling properties

Advanced Oxidation Protein Products-Modified Albumin Induces Differentiation of RAW264.7 Macrophages into Dendritic-Like Cells Which Is Modulated by Cell Surface Thiols.

Local accumulation of Advanced Oxidation Protein Products (AOPP) induces pro-inflammatory and pro-fibrotic processes in kidneys and is an independent predictor of renal fibrosis and of rapid decline of eGFR in patients with chronic kidney disease (CKD). In addition to kidney damage, circulating AOPP may be regarded as mediators of systemic oxidative stress and, in this capacity, they might play a role in the progression of atherosclerotic damage of arterial walls. Atherosclerosis is a chronic inflammatory disease that involves activation of innate and adaptive immunity. Dendritic cells (DCs) are key cells in this process, due to their role in antigen presentation, inflammation resolution and T cell activation. AOPP consist in oxidative modifications of proteins (such as albumin and fibrinogen) that mainly occur through myeloperoxidase (MPO)-derived hypochlorite (HOCl). HOCl modified proteins have been found in atherosclerotic lesions. The oxidizing environment and the shifts in cellular redox equilibrium trigger inflammation, activate immune cells and induce immune responses. Thus, surface thiol groups contribute to the regulation of immune functions. The aims of this work are: (1) to evaluate whether AOPP-proteins induce activation and differentiation of mature macrophages into dendritic cells in vitro; and (2) to define the role of cell surface thiol groups and of free radicals in this process. AOPP-proteins were prepared by in vitro incubation of human serum albumin (HSA) with HOCl. Mouse macrophage-like RAW264.7 were treated with various concentrations of AOPP-HSA with or without the antioxidant N-acetyl cysteine (NAC). Following 48 h of HSA-AOPP treatment, RAW264.7 morphological changes were evaluated by microscopic observation, while markers of dendritic lineage and activation (CD40, CD86, and MHC class II) and allogeneic T cell proliferation were evaluated by flow cytometry. Cell surface thiols were measured by AlexaFluor-maleimide binding, and ROS production was assessed as DCF fluorescence by flow cytometry. HSA-AOPP induced the differentiation of RAW264.7 cells into a dendritic-like phenotype, as shown by morphological changes, by increased CD40, CD86 and MHC class II surface expression and by induction of T cell proliferation. The cell surface thiols dose dependently decreased following HSA-AOPP treatment, while ROS production increased. NAC pre-treatment enhanced the amount of cell surface thiols and prevented their reduction due to treatment with AOPP. Both ROS production and RAW264.7 differentiation into DC-like cells induced by HSA-AOPP were reduced by NAC. Our results highlight that oxidized plasma proteins modulate specific immune responses of macrophages through a process involving changes in the thiol redox equilibrium. We suggest that this mechanism may play a role in determining the rapid progression of the atherosclerotic process observed in CKD patients

Biological heterogeneity of putative bladder cancer stem-like cell populations from human bladder transitional cell carcinoma samples.

Transitional cell carcinoma (TCC) is the most common type of bladder cancer. Emerging evidence has suggested that the capability of a tumor to grow and propagate is dependent on a small subset of cells, the cancer stem-like cells (CSCs) or tumor initiating cells. We report on the isolation and biological characterization of putative bladder CSC populations from primary TCCs. Isolated cells were induced to proliferate in stem cell culture conditions (serum-free medium containing mitogenic growth factors). The proliferating cells formed spheroids (urospheres) and their abilities for extensive proliferation and self-renewal were assayed. Their positivity for several stem cell markers (CD133, Oct-3/4, nestin, and cytokeratins) was also assessed by immunofluorescence tests and they could have the potential to differentiate in the presence of serum. In stem cell culture conditions they gradually showed loss of proliferation, adherence to the substrate, and morphological changes, which might reflect their progressive acquisition of differentiative capacity and loss of self-renewal ability. To evaluate if effective cell selection occurred after isolation, conventional cytogenetic studies on fresh chromosome spreads immediately after isolation and after culture were carried out. In addition, a molecular cytogenetic study by UroVysion assay was carried out on paraffin-embedded tissue sections and on fresh and after culture nuclei preparations. The data collected indicated important karyotype changes and a positive selection for hypo- or near-diploid cells, losing the complexity present in fresh tumors

H2O2 stress damage is reversed by melatonin in a spinal cord organotypic model

Spinal cord injury (SCI) is characterized to be a two-step process: the primary lesion consisting of the initial trauma; the secondary damage, characterized by multiple processes including inflammation, oxidative stress and cell death that lead to a significant expansion of the original damage and to an increase of the functional deficit (1). Among the aforementioned processes, the oxidative stress plays a significant role in pathophysiology of SCI. In this study, we evaluated the role of the melatonin, an indoleamine recognized as a potent antioxidant and immunomodulator (2, 3 )Reiter et al., 1995, Favero et al., 2015), on the oxidative stress, the tissue vitality and the neuritic plasticity in an experimental model of organotypic cultures of Sprague Dawley rat spinal cord slice (SPS) treated with hydrogen peroxide (H2O2) and/or melatonin. Five experimental protocols were performed: 1) control; 2) H2O2 exposure (50 μM); 3) melatonin treatment (5-10M for 24 hours); 4) H2O2 exposure and post-treatment with melatonin; 5) H2O2 exposure after pre-treatment with melatonin. Cellular death was investigated by propidium iodide (PI) assay and the vitality by MTT assay. The total thiols (SH) levels, contrasting the oxidative stress, the neuronal specific nuclear protein (NeuN) and the synaptophysin (Syp) immunopositivity were also evaluated. Melatonin significantly decreases the number of dead cells and increases slice vitality, mainly in slices treated before H2O2 exposure. Moreover, melatonin attenuates total thiols decrease and NeuN and Syp immunopositivity reduction. Overall, these findings suggest that melatonin may exert a potential beneficial effect upon the progression of SCI secondary damage, protecting the tissue from a further degeneration.This work was supported by grants from Giorgio Brunelli Foundation for Spinal Cord Injuries Research