Effect of monomer treatment and polymerisation methods on the bond strength of resin teeth to denture base material

Background: The fracture between acrylic denture base material and artificial teeth is a common clinical occurrence in dental prosthodontic practice. Objective: To evaluate the bond strength between acrylic resins and resin denture teeth when submitted by two protocols of monomer liquid application on the tooth surface and using different polymerisation methods. Material and methods: Microwave‐polymerised (Onda‐Cryl), heat‐polymerised (Clássico) and autopolymerising (Jet) acrylic resins and a brand of resin denture teeth (Biotone) were used. The acrylic resins were polymerised according to the cycles: (A) microwave – fast cycle, Onda‐Cryl; (B) microwave – long cycle, Onda‐Cryl; (C) microwave – manufacturer’s cycle, Onda‐Cryl; (T) water bath – long cycle, Clássico and (Q) bench polymerisation cycle, Jet. Thirty specimens were prepared for each polymerisation method; 10 were packed with acrylic resin after 60 s of monomer liquid application on the tooth surface, 10 after 180 s and 10 without any monomer liquid application. For the purpose of the study, a shear test was used. anova and Tukey tests were performed to identify significant differences (α = 0.05). Results: The highest bond strength values were found for monomer surface treatments, regardless of the polymerisation cycles. The highest significant values were found for cycles B (15.4 ± 1.8 MPa), C (11.9 ± 4.9 MPa) and T (15.4 ± 2.6 MPa) for non‐treated and 60 s methylmethacrylate treated groups. Comparing the monomer liquid treatment, they did not differ significantly (p > 0.05), except for cycle A (p < 0.05). Conclusion: Chemical treatment using monomer on the tooth surface prior to the acrylic resin packing improved the bond strength between resin denture tooth and acrylic resin, regardless of monomer liquid treatment protocols. The microwavable resin, polymerised by fast cycle and autopolymerising resin should be avoided for processing denture and denture repairs, respectively.     

Causes of shear sensitivity of the toxic dinoflagellate Protoceratium reticulatum

Dinoflagellates have proven extremely difficult to culture because they are inhibited by low‐level shear forces. Specific growth rate of the toxic dinoflagellate Protoceratium reticulatum was greatly decreased compared with static control culture by intermittent exposure to a turbulent hydrodynamic environment with a bulk average shear rate that was as low as 0.3 s^?1. Hydrodynamic forces appeared to induce the production of reactive oxygen species (ROS) within the cells and this caused peroxidation of cellular lipids and ultimately cell damage. Exposure to damaging levels of shear rate correlated with the elevated level of lipoperoxides in the cells, but ROS levels measured directly by flow cytometry did not correlate with shear induced cell damage. This was apparently because the measured level of ROS could not distinguish between the ROS that are normally generated by photosynthesis and the additional ROS produced as a consequence of hydrodynamic shear forces. Continuously subjecting the cells to a bulk average shear rate value of about 0.3 s^?1 for 24‐h caused an elevation in the levels of chlorophyll a, peridinin and dinoxanthin, as the cells apparently attempted to counter the damaging effects of shear fields by producing pigments that are potential antioxidants. In static culture, limitation of carbon dioxide produced a small but measureable increase in ROS. The addition of ascorbic acid (0.1 mM) to the culture medium resulted in a significant protective effect on lipid peroxidation, allowing cells to grow under damaging levels of shear rates. This confirmed the use of antioxidant additives as an efficient strategy to counter the damaging effects of turbulence in photobioreactors where shear sensitive dinoflagellates are cultivated. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

Ultra scale‐down studies of the effect of shear on cell quality; Processing of a human cell line for cancer vaccine therapy

Whole cell therapy is showing potential in the clinic for the treatment of many chronic diseases. The translation of laboratory‐scale methods for cell harvesting and formulation to commercial‐scale manufacturing offers major bioprocessing challenges. This is especially the case when the cell properties determine the final product effectiveness. This study is focused on developing an ultra scale‐down method for assessing the impact of the hydrodynamic environment on human cells that constitute the therapeutic product. Small volumes of a prostate cancer cell line, currently being developed in late phase II clinical trials as an allogeneic whole cell vaccine therapy for prostate cancer, were exposed to hydrodynamic shear rates similar to those present in downstream process, formulation and vial filling operations. A small scale rotating disc shear device (20 mL) was used over a range of disc speeds to expose cells to maximum shear rates ranging from 90 × 10³ to 175 × 10³ s^‐1 (equivalent maximum power dissipation rates of 14 × 10³ to 52 × 10³ W kg^‐1). These cells were subsequently analyzed for critical cell quality attributes such as the retention of membrane integrity and cell surface marker profile and density. Three cell surface markers (CD9, CD147, and HLAA‐C) were studied. The cell markers exhibited different levels of susceptibility to hydrodynamic shear but in all cases this was less than or equal to the loss of membrane integrity. It is evident that the marker, or combination or markers, which might provide the required immunogenic response, will be affected by hydrodynamic shear environment during bioprocessing, if the engineering environment is not controlled to within the limits tolerated by the cell components. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

“海棠”台风气流场对褐飞虱北迁路径的影响

基于GIS、GrADS软件和HYSPLIT 4.8轨迹模式,分析了0505号台风“海棠”发生期间（2005年7月19—21日）中国10个省42个虫情观测点的逐日灯诱褐飞虱虫量、850 hPa等压面的风场和20个虫情监测点的褐飞虱迁飞轨迹.结果表明：台风“海棠”登陆中国后,改变了引导褐飞虱向北迁飞的西南气流,造成风场在台风西南部的辐合和大范围的转向,阻止了褐飞虱的向北迁飞,迫使其在某些区域集中迫降;850 hPa等压面上切变线附近是褐飞虱集中降落的区域;在台风衰亡时期,台风东南部气流暖式切变区是大量降虫的区域;台风整体登陆后,西南气流的再次建立,造成褐飞虱的大量北迁.     

Down-regulation of neprilysin (EC3.4.24.11) expression in vascular endothelial cells by laminar shear stress involves NADPH oxidase-dependent ROS production

Neprilysin (NEP, neutral endopeptidase, EC3.4.24.11), a zinc metallopeptidase expressed on the surface of endothelial cells, influences vascular homeostasis primarily through regulated inactivation of natriuretic peptides and bradykinin. Earlier in vivo studies reporting on the anti-atherosclerotic effects of NEP inhibition and on the atheroprotective effects of flow-associated laminar shear stress (LSS) have lead us to hypothesize that the latter hemodynamic stimulus may serve to down-regulate NEP levels within the vascular endothelium. To address this hypothesis, we have undertaken an investigation of the effects of LSS on NEP expression in vitro in bovine aortic endothelial cells (BAECs), coupled with an examination of the signalling mechanism putatively mediating these effects. BAECs were exposed to physiological levels of LSS (10 dynes/cm², 24 h) and harvested for analysis of NEP expression using real-time PCR, Western blotting, and immunocytochemistry. Relative to unsheared controls, NEP mRNA and protein were substantially down-regulated by LSS (≥50%), events which could be prevented by treatment of BAECs with either N-acetylcysteine, superoxide dismutase, or catalase, implicating reactive oxygen species (ROS) involvement. Employing pharmacological and molecular inhibition strategies, the signal transduction pathway mediating shear-dependent NEP suppression was also examined, and roles implicated for Gβγ, Rac1, and NADPH oxidase activation in these events. Treatment of static BAECs with angiotensin-II, a potent stimulus for NADPH oxidase activation, mimicked the suppressive effects of shear on NEP expression, further supporting a role for NADPH oxidase-dependent ROS production. Interestingly, inhibition of receptor tyrosine kinase signalling had no effect. In conclusion, we confirm for the first time that NEP expression is down-regulated in vascular endothelial cells by physiological laminar shear, possibly via a mechanotransduction mechanism involving NADPH oxidase-induced ROS production.     

Effect of LDL concentration polarization on the uptake of LDL by human endothelial cells and smooth muscle cells co-cultured

To substantiate our hypothesis that concentration polarization of low-density Upoprotein （LDL） plays an important role in the localization of atherogenesis, we investigated the effects of wall shear stress and water filtration rate （or perfusion pressure） on the luminal surface LDL concentration （cw） and the LDL uptake by human vascular endothelial cells and smooth muscle cells co-cultured on a permeable membrane using a parallel-plate flow chamber technique and a flow cyto-metry method. The results indicated that the uptake of fluorescent labeled LDL （DiI-LDL） by the co-cultured cells was positively correlated with Cw in a non-linear fashion. When cw was low, the uptake increased very sharply with increasing Cw. Then the increase became gradual and the uptake was seemingly leveled out when Cw reached beyond 160 μg/ml. The present study therefore has provided further experimental evidence that concentration polarization may occur in the arterial system and have a positive correlation with the uptake of LDLs by the arterial wall, which gives support to our hypothesis regarding the localization of atherogenesis.     

Application of Numerical Methods to Elasticity Imaging

Elasticity imaging can be understood as the intersection of the study of biomechanical properties, imaging sciences, and physics. It was mainly motivated by the fact that pathological tissue presents an increased stiffness when compared to surrounding normal tissue. In the last two decades, research on elasticity imaging has been an international and interdisciplinary pursuit aiming to map the viscoelastic properties of tissue in order to provide clinically useful information. As a result, several modalities of elasticity imaging, mostly based on ultrasound but also on magnetic resonance imaging and optical coherence tomography, have been proposed and applied to a number of clinical applications: cancer diagnosis (prostate, breast, liver), hepatic cirrhosis, renal disease, thyroiditis, arterial plaque evaluation, wall stiffness in arteries, evaluation of thrombosis in veins, and many others. In this context, numerical methods are applied to solve forward and inverse problems implicit in the algorithms in order to estimate viscoelastic linear and nonlinear parameters, especially for quantitative elasticity imaging modalities. In this work, an introduction to elasticity imaging modalities is presented. The working principle of qualitative modalities (sonoelasticity, strain elastography, acoustic radiation force impulse) and quantitative modalities (Crawling Waves Sonoelastography, Spatially Modulated Ultrasound Radiation Force (SMURF), Supersonic Imaging) will be explained. Subsequently, the areas in which numerical methods can be applied to elasticity imaging are highlighted and discussed. Finally, we present a detailed example of applying total variation and AM-FM techniques to the estimation of elasticity.     

Signaling mechanisms in red blood cells: A view through the protein phosphorylation and deformability

Intracellular signaling mechanisms in red blood cells (RBCs) involve various protein kinases and phosphatases and enable rapid adaptive responses to hypoxia, metabolic requirements, oxidative stress, or shear stress by regulating the physiological properties of the cell. Protein phosphorylation is a ubiquitous mechanism for intracellular signal transduction, volume regulation, and cytoskeletal organization in RBCs. Spectrin-based cytoskeleton connects integral membrane proteins, band 3 and glycophorin C to junctional proteins, ankyrin and Protein 4.1. Phosphorylation leads to a conformational change in the protein structure, weakening the interactions between proteins in the cytoskeletal network that confers a more flexible nature for the RBC membrane. The structural organization of the membrane and the cytoskeleton determines RBC deformability that allows cells to change their ability to deform under shear stress to pass through narrow capillaries. The shear stress sensing mechanisms and oxygenation-deoxygenation transitions regulate cell volume and mechanical properties of the membrane through the activation of ion transporters and specific phosphorylation events mediated by signal transduction. In this review, we summarize the roles of Protein kinase C, cAMP-Protein kinase A, cGMP-nitric oxide, RhoGTPase, and MAP/ERK pathways in the modulation of RBC deformability in both healthy and disease states. We emphasize that targeting signaling elements may be a therapeutic strategy for the treatment of hemoglobinopathies or channelopathies. We expect the present review will provide additional insights into RBC responses to shear stress and hypoxia via signaling mechanisms and shed light on the current and novel treatment options for pathophysiological conditions.     

Arterial Shear Stress Reduces Eph-B4 Expression in Adult Human Veins

Vein graft adaptation to the arterial environment is characterized by loss of venous identity, with reduced Ephrin type-B receptor 4 (Eph-B4) expression but without increased Ephrin-B2 expression. We examined changes of vessel identity of human saphenous veins in a flow circuit in which shear stress could be precisely controlled. Medium circulated at arterial or venous magnitudes of laminar shear stress for 24 hours; histologic, protein, and RNA analyses of vein segments were performed. Vein endothelium remained viable and functional, with platelet endothelial cell adhesion molecule (PECAM)-expressing cells on the luminal surface. Venous Eph-B4 expression diminished (p = .002), Ephrin-B2 expression was not induced (p = .268), and expression of osteopontin (p = .002) was increased with exposure to arterial magnitudes of shear stress. Similar changes were not found in veins placed under venous flow or static conditions. These data show that human saphenous veins remain viable during ex vivo application of shear stress in a bioreactor, without loss of the venous endothelium. Arterial magnitudes of shear stress cause loss of venous identity without gain of arterial identity in human veins perfused ex vivo. Shear stress alone, without immunologic or hormonal influence, is capable of inducing changes in vessel identity and, specifically, loss of venous identity.