Substrate stiffness promotes vascular smooth muscle cell calcification by reducing the levels of nuclear actin monomers.

BACKGROUND: Vascular calcification (VC) is a prevalent independent risk factor for adverse cardiovascular events and is associated with diabetes, hypertension, chronic kidney disease, and atherosclerosis. However, the mechanisms regulating the osteogenic differentiation of vascular smooth muscle cells (VSMC) are not fully understood. METHODS: Using hydrogels of tuneable stiffness and lysyl oxidase-mediated stiffening of human saphenous vein ex vivo, we investigated the role of substrate stiffness in the regulation of VSMC calcification. RESULTS: We demonstrate that increased substrate stiffness enhances VSMC osteogenic differentiation and VSMC calcification. We show that the effects of substrate stiffness are mediated via a reduction in the level of actin monomer within the nucleus. We show that in cells interacting with soft substrate, elevated levels of nuclear actin monomer repress osteogenic differentiation and calcification by repressing YAP-mediated activation of both TEA Domain transcription factor (TEAD) and RUNX Family Transcription factor 2 (RUNX2). CONCLUSION: This work highlights for the first time the role of nuclear actin in mediating substrate stiffness-dependent VSMC calcification and the dual role of YAP-TEAD and YAP-RUNX2 transcriptional complexes.

Effect of Finishing and Polishing on Roughness and Gloss of Lithium Disilicate and Lithium Silicate Zirconia Reinforced Glass Ceramic for CAD/CAM Systems.

OBJECTIVE: To assess the efficacy of dedicated finishing/polishing systems on roughness and gloss of VITA Suprinity and IPS e.max CAD. METHOD: A total of 24 blocks of Suprinity and 24 of e.max were cut into a wedge shape using an InLab MC-XL milling unit. After crystallization, the 24 Suprinity wedges were divided into four subgroups: group A.1: Suprinity Polishing Set Clinical used for 30 seconds and group A.2: for 60 seconds; group A.3: VITA Akzent Plus Paste; and group A.4: spray. The 24 e.max wedges (group B) were divided into four subgroups according to the finishing procedure: group B.1: Optrafine Ceramic Polishing System for 30 seconds and group B.2: for 60 seconds; group B.3: IPS e.max CAD Crystall/Glaze paste; and group B.4: spray. After finishing/polishing, gloss was assessed with a glossmeter and roughness evaluated with a profilometer. Results were analyzed by applying a two-way analysis of variance for gloss and another for roughness (α=0.05). One specimen per each subgroup was observed with a scanning electron microscope. RESULTS: For roughness, materials and surface were significant factors ( p<0.001). Suprinity exhibited significantly lower roughness than e.max. Also the Material-Surface Treatment interaction was statistically significant ( p=0.026). For gloss, both material and surface treatment were significant factors ( p<0.001). VITA Suprinity showed significantly higher gloss than e.max. Also the Material-Surface Treatment interaction was statistically significant ( p<0.001). CONCLUSIONS: Manual finishing/polishing for 60 seconds and glazing paste are the most effective procedures in lowering the roughness of CAD/CAM silica-based glass ceramics. Manual finishing/polishing for 60 seconds allows milled silica-based glass ceramics to yield a higher gloss. VITA Suprinity displayed higher polishability than IPS e.max CAD.

Post-Retained Single Crowns versus Fixed Dental Prostheses: A 7-Year Prospective Clinical Study.

Biomechanical integrity of endodontically treated teeth (ETT) is often compromised. Degree of hard tissue loss and type of final prosthetic restoration should be carefully considered when making a treatment plan. The objective of this prospective clinical trial was to assess the influence of the type of prosthetic restoration as well as the degree of hard tissue loss on 7-y clinical performance of ETT restored with fiber posts. Two groups ( n = 60) were defined depending on the type of prosthetic restoration needed: 1) single unit porcelain-fused-to-metal (PFM) crowns (SCs) and 2) 3- to 4-unit PFM fixed dental prostheses (FDPs), with 1 healthy and 1 endodontically treated and fiber post-restored abutment. Within each group, samples were divided into 2 subgroups ( n = 30) according to the amount of residual coronal tissues after abutment buildup and final preparation: A) >50% of coronal residual structure or B) equal to or <50% of coronal residual structure. The clinical outcome was assessed based on clinical and intraoral radiographic examinations at the recalls after 6, 12, 24, 36, 48, and 84 mo. Data were analyzed by Kaplan-Meier log-rank test and Cox regression analysis ( P < 0.05). The overall 7-y survival rate of ETT restored with fiber post and either SCs or FDPs was 69.2%. The highest 84-mo survival rate was recorded in group 1A (90%), whereas teeth in group 2B exhibited the lowest performance (56.7% survival rate). The log-rank test detected statistically significant differences in survival rates among the groups ( P = 0.048). Cox regression analysis revealed that the amount of residual coronal structure ( P = 0.041; hazard ratio [HR], 2.026; 95% confidence interval [CI] for HR, 1.031-3.982) and the interaction between the type of prosthetic restoration and the amount of residual coronal structure ( P = 0.024; HR, 1.372; 95% CI for HR, 1.042-1.806) were statistically significant factors for survival ( ClinicalTrials.gov NCT01532947).

Effect of Finishing and Polishing on the Surface Roughness and Gloss of Feldspathic Ceramic for Chairside CAD/CAM Systems.

OBJECTIVES: To evaluate surface roughness and gloss of feldspathic ceramic blocks for chairside CAD/CAM systems before and after finishing and polishing. METHODS: VITA Mark II ceramic blocks for the CEREC CAD/CAM system were cut perpendicularly in order to obtain a total of 70 specimens (14 × 18 × 3 mm). The flat surface was roughened using a grinder/polisher with dry 120-grit silicone-carbide paper. Surface roughness and gloss were measured using a digital profilometer (Ra) and a glossmeter (GU), respectively. Specimens were randomly divided into seven groups (n=10) based on the finishing/polishing system as follows: 1) Identoflex NGPorcelain Polisher (INP), 2) Identoflex Diamond Ceramic Polisher (IDP), 3) Hiluster Polishing System (HPS), 4) OptraFine (OF), 5) Identoflex Lucent (IL), 6) VITA Akzent Glaze Spray (AGS), and 7) VITA Shading Paste and Liquid (SPL). Surface analysis was repeated after the finishing/polishing treatment, and the obtained data were compared to the baseline in order to evaluate the ΔRa and ΔGU. Results were statistically analyzed. The surface morphology was observed by scanning electron microscopy. RESULTS: The mean surface roughness of polished systems increased in the order (statistical groups designated) SPLa < ILa < OFab < IDPbc < AGSbc < INPbc < HPSc and mean gloss decreased in the order AGSa > SPLa > OFab > ILabc > HPSbcd > INPcd > IDPd. CONCLUSIONS: The smoothest surface of CAD/CAM feldspathic ceramic blocks was achieved using the furnace-based glaze systems VITA Akzent Glaze Spray and VITA Shading Paste and Liquid and manual systems Identoflex Lucent and OptraFine.

Control of infectious mortality due to carbapenemase-producing Klebsiella pneumoniae in hematopoietic stem cell transplantation.

Carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) infections are an emerging cause of death after hematopoietic stem cell transplantation (HSCT). In allogeneic transplants, mortality rate may rise up to 60%. We retrospectively evaluated 540 patients receiving a transplant from an auto- or an allogeneic source between January 2011 and October 2015. After an Institutional increase in the prevalence of KPC-Kp bloodstream infections (BSI) in June 2012, from July 2012, 366 consecutive patients received the following preventive measures: (i) weekly rectal swabs for surveillance; (ii) contact precautions in carriers (iii) early-targeted therapy in neutropenic febrile carriers. Molecular typing identified KPC-Kp clone ST512 as the main clone responsible for colonization, BSI and outbreaks. After the introduction of these preventive measures, the cumulative incidence of KPC-Kp BSI (P=0.01) and septic shocks (P=0.01) at 1 year after HSCT was significantly reduced. KPC-Kp infection-mortality dropped from 62.5% (pre-intervention) to 16.6% (post-intervention). Day 100 transplant-related mortality and KPC-Kp infection-related mortality after allogeneic HSCT were reduced from 22% to 10% (P=0.001) and from 4% to 1% (P=0.04), respectively. None of the pre-HSCT carriers was excluded from transplant. These results suggest that active surveillance, contact precautions and early-targeted therapies, may efficiently control KPC-Kp spread and related mortality even after allogeneic HSCT.

Design, fabrication and perivascular implantation of bioactive scaffolds engineered with human adventitial progenitor cells for stimulation of arteriogenesis in peripheral ischemia.

Cell therapy represents a promising option for revascularization of ischemic tissues. However, injection of dispersed cells is not optimal to ensure precise homing into the recipient's vasculature. Implantation of cell-engineered scaffolds around the occluded artery may obviate these limitations. Here, we employed the synthetic polymer polycaprolactone for fabrication of 3D woodpile- or channel-shaped scaffolds by a computer-assisted writing system (pressure assisted micro-syringe square), followed by deposition of gelatin (GL) nanofibers by electro-spinning. Scaffolds were then cross-linked with natural (genipin, GP) or synthetic (3-glycidyloxy-propyl-trimethoxy-silane, GPTMS) agents to improve mechanical properties and durability in vivo. The composite scaffolds were next fixed by crown inserts in each well of a multi-well plate and seeded with adventitial progenitor cells (APCs, 3 cell lines in duplicate), which were isolated/expanded from human saphenous vein surgical leftovers. Cell density, alignment, proliferation and viability were assessed 1 week later. Data from in vitro assays showed channel-shaped/GPTMS-crosslinked scaffolds confer APCs with best alignment and survival/growth characteristics. Based on these results, channel-shaped/GPTMS-crosslinked scaffolds with or without APCs were implanted around the femoral artery of mice with unilateral limb ischemia. Perivascular implantation of scaffolds accelerated limb blood flow recovery, as assessed by laser Doppler or fluorescent microspheres, and increased arterial collaterals around the femoral artery and in limb muscles compared with non-implanted controls. Blood flow recovery and perivascular arteriogenesis were additionally incremented by APC-engineered scaffolds. In conclusion, perivascular application of human APC-engineered scaffolds may represent a novel option for targeted delivery of therapeutic cells in patients with critical limb ischemia.

Effect of repeated firings on flexural strength of veneered zirconia.

OBJECTIVE: Chipping and/or delamination represent a clinical failure of porcelain fused to zirconia (PFZ) prostheses. Causes and solutions have not been completely clarified. The present study was aimed at evaluating the effects of number of firings on the flexural strength of PFZ specimen. METHODS: Forty-five zirconia specimens in shape of bars were cut, sintered and divided in 3 groups (n=15). Group 1: veneering ceramic was layered "in bulk" and fired. Group 2: veneering ceramic was layered in three layers, individually fired. Group 3: veneering ceramic was layered in five layers, individually fired. Each layer thickness was controlled by the use of calibrated molds. The total veneering ceramic thickness for all the specimens was 1.2mm, and the total thickness of the specimen of 2.0mm. Three-point bending test was performed. Fracture load was recorded in Newton and MPa value was calculated taking into account the bi-layered nature of the specimen. Data were statistically analyzed. RESULTS: Specimens obtained with on single firing cycle obtained a statistically significant (p<0.001) lower flexural strength (54.61±8.98MPa) than specimens veneered with 3 or 5 firing cycles. The last two obtained very similar results (77.63±13.17MPa and 73.62±12.38MPa respectively) and the differences was not statistically significant. SIGNIFICANCE: In bi-layered PFZ specimen, three to five layers and firings determine higher flexural resistance when compared to a single firing. Thus, a 3-layers veneering procedure is recommended to increase flexural resistance. If a 5-layer procedure is necessary to improve esthetics, it does not decrease flexural resistance.

Incidence, risk factors and clinical outcome of leukemia relapses with loss of the mismatched HLA after partially incompatible hematopoietic stem cell transplantation.

Genomic loss of the mismatched human leukocyte antigen (HLA) is a recently described mechanism of leukemia immune escape and relapse after allogeneic hematopoietic stem cell transplantation (HSCT). Here we first evaluated its incidence, risk factors and outcome in 233 consecutive transplants from partially HLA-mismatched related and unrelated donors (MMRD and MMUD, respectively). We documented 84 relapses, 23 of which with HLA loss. All the HLA loss relapses occurred after MMRD HSCT, and 20/23 in patients with acute myeloid leukemia. Upon MMRD HSCT, HLA loss variants accounted for 33% of the relapses (23/69), occurring later than their 'classical' counterparts (median: 307 vs 88 days, P<0.0001). Active disease at HSCT increased the risk of HLA loss (hazard ratio (HR): 10.16; confidence interval (CI): 2.65-38.92; P=0.001), whereas older patient ages had a protective role (HR: 0.16; CI: 0.05-0.46; P=0.001). A weaker association with HLA loss was observed for graft T-cell dose and occurrence of chronic graft-versus-host disease. Outcome after 'classical' and HLA loss relapses was similarly poor, and second transplantation from a different donor appeared to provide a slight advantage for survival. In conclusion, HLA loss is a frequent mechanism of evasion from T-cell alloreactivity and relapse in patients with myeloid malignancies transplanted from MMRDs, warranting routine screening in this transplantation setting.

Sirolimus-based graft-versus-host disease prophylaxis promotes the in vivo expansion of regulatory T cells and permits peripheral blood stem cell transplantation from haploidentical donors.

Hematopoietic stem cell transplantation (HSCT) from human leukocyte antigen (HLA) haploidentical family donors is a promising therapeutic option for high-risk hematologic malignancies. Here we explored in 121 patients, mostly with advanced stage diseases, a sirolimus-based, calcineurin-inhibitor-free prophylaxis of graft-versus-host disease (GvHD) to allow the infusion of unmanipulated peripheral blood stem cell (PBSC) grafts from partially HLA-matched family donors (TrRaMM study, Eudract 2007-5477-54). Conditioning regimen was based on treosulfan and fludarabine, and GvHD prophylaxis on antithymocyte globulin Fresenius (ATG-F), rituximab and oral administration of sirolimus and mycophenolate. Neutrophil and platelet engraftment occurred in median at 17 and 19 days after HSCT, respectively, and full donor chimerism was documented in patients' bone marrow since the first post-transplant evaluation. T-cell immune reconstitution was rapid, and high frequencies of circulating functional T-regulatory cells (Treg) were documented during sirolimus prophylaxis. Incidence of acute GvHD grade II-IV was 35%, and occurrence and severity correlated negatively with Treg frequency. Chronic GvHD incidence was 47%. At 3 years after HSCT, transpant-related mortality was 31%, relapse incidence 48% and overall survival 25%. In conclusion, GvHD prophylaxis with sirolimus-mycophenolate-ATG-F-rituximab promotes a rapid immune reconstitution skewed toward Tregs, allowing the infusion of unmanipulated haploidentical PBSC grafts.

Early molecular diagnosis of aspergillosis in a patient with acute myeloid leukaemia.

Diagnosis of invasive fungal infection remains challenging. Here we report a case of early diagnosis of invasive aspergillosis in a neutropenic patient affected by acute myeloid leukaemia, achieved through the detection of Aspergillus fumigatus species-specific ribonucleic acid sequences by a sensitive multiplex real-time polymerase chain reaction-based molecular assay. Thanks to the early diagnosis, targeted therapy was promptly established and the severe fungal infection controlled, allowing the patient to subsequently receive allogeneic hematopoietic stem cell transplantation from a haploidentical donor, her only curative option. Also in this instance, targeted secondary antifungal prophylaxis with voriconazole avoided any other fungal infection afterwards. This report suggests how the implementation of molecular assays in combination with routine diagnostic procedures, can improve microbiological diagnosis in sepsis, particularly in case of fungal infection, difficult to detect with standard microbiological culture methods.

Bond strength to unground enamel and sealing ability in pits and fissures of a new self-adhering flowable resin composite.

OBJECTIVE: To evaluate the applicability as a sealant of a new self-adhering flowable resin composite (Vertise Flow, Kerr, VF) by assessing shear bond strength (SBS) to unground enamel and microleakage (microLKG) in sealed pits and fissures. STUDY DESIGN: Marketed sealants to be used in combination with phosphoric acid (Guardian Seal, Kerr, GS) or with a self-etch adhesive (Adper Prompt-L-Pop/Clinpro Sealant, 3M ESPE, CS) were compared to VF. For SBS testing on unground enamel 10 molars per group were used. For microLKG assessment, pits and fissures sealing was performed in 12 molars per group. The sealed teeth were immersed in a 50% weight silver nitrate solution for 24 hours and the extent of interfacial leakage was measured. Between-group differences in SBS were assessed using One-Way Analysis of Variance (ANOVA), followed by Tukey test (p < 0.05). microLKG data were analyzed with Kruskall-Wallis ANOVA (p > 0.05). RESULTS: SBS of VF was statistically similar to that measured by CS and higher than that of GS. Interfacial leakage was similar in the three groups. CONCLUSIONS: The finding of satisfactory bond strength and sealing ability of VF when compared to the marketed sealants encourages the use of VF in pit and fissure sealing.

Color correspondence of different batches of TriLuxe CAD/CAM ceramic blocks.

TriLuxe ceramic blocks for chairside CAD/CAM procedures are color layered to allow natural esthetics, and only require subsequent glazing. The purpose of this study was to compare color repeatability of different batches of TriLuxe blocks. The three commercially available shades (1M2C, 2M2C, 3M2C) of TriLuxe blocks for the Cerec CAD/CAM system were examined. For each of the three colors, three different batches were tested, 5 blocks each. The measurements were made using a spectrophotometer equipped with an integrating sphere using the CIELab* colorimetric system. One-way ANOVA showed that the factor "Production Batch" was not statistically significant. Regarding deltaE, none of the 315 color comparisons (neither within the same shade, nor between specimens of the same batch, nor between specimens from different batches) exceeded the proposed deltaE = 3.3 threshold for clinical acceptability. All the different batches of the different shades of VITA TriLuxe blocks for the Cerec system showed the high degree of color correspondence necessary in industrially prefabricated CAD/CAM blocks.

In etanercept-treated psoriatic arthritis patients clinical improvement correlated with an increase of serum cortisol relative to other adrenal hormones.

OBJECTIVE: In patients with rheumatoid arthritis (RA), long-term therapy with anti-tumor necrosis factor (TNF) antibodies sensitizes the pituitary gland and improves adrenal androgen secretion in prednisolone-naïve patients. However, whether this is similar in psoriatic arthritis (PsA) is not known. The aim of this study was to assess the effect of 12 weeks of etanercept treatment upon the function of the HPA axis in patients with PsA. METHODS: Eleven prednisolone-naïve patients (mean age 47.3+/-8.9 years) with PsA were included. We measured serum levels of adrenocorticotropic hormone (ACTH), 17-hydroxyprogesterone (17OHP), cortisol, and androstenedione (ASD), at baseline and at 4 and 12 weeks after initiation of anti-TNF therapy (etanercept, 50 mg every week as a single dose by sc. injection). Clinical improvement was assessed using the Disease Activity Score-28 (DAS-28). RESULTS: Mean levels of serum ACTH, serum cortisol, serum 17OHP and serum ASD did not markedly change during 12 weeks of etanercept treatment. Similarly, the ratio of serum cortisol divided by serum ACTH did not change during 12 weeks of anti-TNF treatment. However, an increase of serum cortisol relative to serum 17OHP or ASD was related to clinical improvement. This indicates that improvement was linked to higher serum cortisol levels relative to others adrenal hormones. CONCLUSION: This is the first study to demonstrate baseline serum levels and the course of HPA axis-related hormones in patients with PsA. An increase of serum cortisol relative to others adrenocortical hormones (i.e., androstenedione and ACTH) was accompanied by clinical improvement.

Unusual presentation of large B cell lymphoma: a case report and review of literature.

Diffuse large B cell lymphoma (DLBCL) is the largest subtype of non-Hodgkin's lymphomas (NHLs) and is characterized by relatively frequent extranodal presentation. In these cases, the most common extranodal localizations are stomach, CNS, bone, testis and liver. Simultaneous detection of multiple extranodal involvement at presentation is quite uncommon, with the majority of these cases characterized by gastric or intestinal disease localization. Retrospective analysis concerning multifocal extranodal NHLs never pointed out disease features such as those described here. We report a patient with an unusual presentation of DLBCL, characterized by adrenal and renal involvement, associated with symptoms and signs of the cold agglutinin disease and a hypercoagulable state. Subsequently, computed tomography (CT) and fluorodeoxyglucose-positron emission tomography (FDG-PET) scanning disclosed a rapidly extensive spread to nodes and bones. Cytofluorimetric analysis of a renal specimen showed medium-to-large lympho-monocytoid elements positive for CD20 with monoclonal expression of immunoglobulin kappa light chain. Histopathological examination confirmed a renal CD20 positive DLBCL localization.