The thrombin receptor in adrenal medullary microvascular endothelial cells is negatively coupled to adenylyl cyclase through a Gi protein

AbstractThe effects of thrombin on adenylyl cyclase activity were examined in rat adrenal medullary microvascular endothelial cells (RAMEC). Confluent RAMEC monolayers were stimulated for 5 min with cAMP-generating agents in the absence and presence of thrombin, and intracellular cAMP was measured with a radioligand binding assay. Thrombin (0.001–0.25 U/ml) dose-dependently inhibited IBMX-, isoproterenol- and forskolin-stimulated cAMP accumulation. A peptide agonist of the thrombin receptor, γ-thrombin, and the serine proteases trypsin and plasmin, also inhibited agonist-stimulated cAMP levels, while proteolytically inactive PPACK- or DIP-α-thrombins were without effect. Moreover, the thrombin inhibitor hirudin abolished the inhibitory effect of thrombin but not of the peptide agonist. These results suggest that the inhibitory action of thrombin on cAMP accumulation is mediated by a proteolytically-activated thrombin receptor. The inhibitor of Gi-proteins pertussis toxin abolished the inhibitory effect of thrombin on isoproterenol- or IBMX-stimulated cAMP production, while the phorbol ester PMA partly impaired it. The protein kinase C inhibitors staurosporine or H7 and the intracellular Ca2+ chelator BAPTA-AM were without effect. Collectively, our data suggest that the thrombin receptor in RAMEC is negatively coupled to adenylyl cyclase through a pertussis toxin-sensitive Gi-protein

Dynamic hi-resolution horizontal microscope for monitoring and manipulating real-time tissue assembly

Poster presented at Biomedical Technology Showcase 2006, Philadelphia, PA. Retrieved 18 Aug 2006 from http://www.biomed.drexel.edu/new04/Content/Biomed_Tech_Showcase/Poster_Presentations/Lelkes_4.pdf.There is an un-met clinical and technological need in regenerative medicine and tissue engineering for developing a non-invasive technique for monitoring and manipulating tissue assembly. Our goal is to create an integrated hardware/ software system to automatically diagnose, monitor, and control tissue growth in dynamic bioreactors in real-time. For this we propose to engineer and test a versatile, hi-resolution horizontal microscope that can intelligently aid in the on-line optimization of tissue growth parameters within a bioreactor. Having developed a functional prototype, we succeeded in compacting and significantly reducing the costs for the 2nd generation system, which now needs further validation

How Christians Reconcile Their Personal Political Views and the Teachings of Their Faith: Projection as a Means of Dissonance Reduction

The present study explores the dramatic projection of one\u27s own views onto those of Jesus among conservative and liberal American Christians. In a large-scale survey, the relevant views that each group attributed to a contemporary Jesus differed almost as much as their own views. Despite such dissonance-reducing projection, however, conservatives acknowledged the relevant discrepancy with regard to fellowship issues (e.g., taxation to reduce economic inequality and treatment of immigrants) and liberals acknowledged the relevant discrepancy with regard to morality issues (e.g., abortion and gay marriage). However, conservatives also claimed that a contemporary Jesus would be even more conservative than themselves on the former issues whereas liberals claimed that Jesus would be even more liberal than themselves on the latter issues. Further reducing potential dissonance, liberal and conservative Christians differed markedly in the types of issues they claimed to be more central to their faith. A concluding discussion considers the relationship between individual motivational processes and more social processes that may underlie the present findings, as well as implications for contemporary social and political conflict

MICRO CHARACTERIZATION OF MG AND MG ALLOY FOR BIODEGRADABLE ORTHOPEDIC IMPLANTS APPLICATION

ABSTRACT Magnesium as a candidate metallic biomaterial for biodegradable orthopedic implants was evaluated in-vitro in terms of degradation behavior, biocompatibility and mechanical property both in macro-and micro-scale. Micro structure of pure Mg and AZ61 after degradation in both simulated body fluid (SBF) and cell culture environment were analyzed. Different from AZ61, pure Mg degraded at a higher rate and attracted large amount of salt precipitation which formed a layer covering the surface. Much less pitting degradation and salt deposition were observed on both pure Mg and AZ61 in cell culture environment compared to in SBF. After culturing for 7 days, EAhy926 cells growing on AZ61 showed significant higher proliferation rate as of cells growing on pure Mg. Higher proliferation rates indicated that cells grew better on slow-degrading AZ61 than on fast-degrading pure Mg. Cells growing on AZ61 proliferated much better and assembled together to form a consistent tissue-like micro-structure, while cells spread and reached out on the surface of pure Mg, possibly due to low cell density and lack of cellular communication. The elastic modulus and tensile yield strength of magnesium are closer to those of natural bone than other commonly used metallic biomaterials. It was shown that Mg was biodegradable, biocompatible and had appropriate mechanical strength, thus Mg and its alloys showed great potential for deployment in a new generation of biodegradable orthopedic implants

Bone scaffold fabrication using porogen based injection molding method and biocomposite materials

Proceedings of the Seventh International Conference on Frontiers of Design and Manufacturing, in Guangzhou, China, June 20, 2006, 1: pp. 267-272.Drop on demand printing (DDP) is a solid freeform fabrication (SFF) technique capable of generating physical features required for scaffolds to be used in hard tissue repair. Here we report results toward the development of a reproducible manufacturing process for tissue engineering scaffolds based on injectable porogens fabricated by DDP. Thermoplastic porogens were designed using Pro/Engineer and fabricated with a commercially available DDP machine. Scaffolds composed of either pure polycaprolactone (PCL) or homogeneous composites of PCL and calcium phosphate (CaP, 10% or 20% w/w) were subsequently fabricated by injection molding of molten polymer-ceramic composites. The precisely formed scaffolds were separated from the porogens in an agitated ethanol bath. Attainable scaffold pore sizes using the porogen-based method were found to be 200 μm for pure PCL. We characterized the compressive strength of 90:10 and 80:20 PCL-CaP composite materials (19.5+/-1.4 MPa and 24.8+/-1.3MPa respectively) according to ASTM standards, as wells as pure PCL scaffolds (13+/-1.2 MPa) fabricated using our process. Initial cell-biomaterial interaction studies demonstrated that our PCL and 80:20 PCL-CaP composite scaffolds supported attachment and proliferation of human embryonic palatal mesenchymal (HEPM) cells, as evidenced by fluorescent nuclear staining and the Alamar Blue™ assay. Scanning electron microscopy (SEM) revealed that HEPM cells spread and demonstrated histiotypic mesenchymal morphology

Comparisons of depression, anxiety, well-being, and perceptions of the built environment amongst adults seeking social, intermediate and market-rent accommodation in the former London Olympic Athletes' Village.

The Examining Neighbourhood Activities in Built Living Environments in London (ENABLE London) study provides a unique opportunity to examine differences in mental health and well-being amongst adults seeking social, intermediate (affordable rent), and market-rent housing in a purpose built neighbourhood (East Village, the former London 2012 Olympic Athletes' Village), specifically designed to encourage positive health behaviours. Multi-level logistic regression models examined baseline differences in levels of depression, anxiety and well-being across the housing groups. Compared with the intermediate group, those seeking social housing were more likely to be depressed, anxious and had poorer well-being after adjustment for demographic and health status variables. Further adjustments for neighbourhood perceptions suggest that compared with the intermediate group, perceived neighbourhood characteristics may be an important determinant of depression amongst those seeking social housing, and lower levels of happiness the previous day amongst those seeking market-rent housing. These findings add to the extensive literature on inequalities in health, and provide a strong basis for future longitudinal work that will examine change in depression, anxiety and well-being after moving into East Village, where those seeking social housing potentially have the most to gain

Digalactosyl-diacylglycerol-deficiency lowers the thermal stability of thylakoid membranes

We investigated the effects of digalactosyl-diacylglycerol (DGDG) on the organization and thermal stability of thylakoid membranes, using wild-type Arabidopsis thaliana and the DGDG-deficient mutant, dgd1. Circular-dichroism measurements reveal that DGDG-deficiency hampers the formation of the chirally organized macrodomains containing the main chlorophyll a/b light-harvesting complexes. The mutation also brings about changes in the overall chlorophyll fluorescence lifetimes, measured in whole leaves as well as in isolated thylakoids. As shown by time-resolved measurements, using the lipophylic fluorescence probe Merocyanine 540 (MC540), the altered lipid composition affects the packing of lipids in the thylakoid membranes but, as revealed by flash-induced electrochromic absorbance changes, the membranes retain their ability for energization. Thermal stability measurements revealed more significant differences. The disassembly of the chiral macrodomains around 55°C, the thermal destabilization of photosystem I complex at 61°C as detected by green gel electrophoresis, as well as the sharp drop in the overall chlorophyll fluorescence lifetime above 45°C (values for the wild type—WT) occur at 4–7°C lower temperatures in dgd1. Similar differences are revealed in the temperature dependence of the lipid packing and the membrane permeability: at elevated temperatures MC540 appears to be extruded from the dgd1 membrane bilayer around 35°C, whereas in WT, it remains lipid-bound up to 45°C and dgd1 and WT membranes become leaky around 35 and 45°C, respectively. It is concluded that DGDG plays important roles in the overall organization of thylakoid membranes especially at elevated temperatures

Permian high-temperature metamorphism in the Western Alps (NW Italy)

During the late Palaeozoic, lithospheric thinning in part of the Alpine realm caused high-temperature low-to-medium pressure metamorphism and partial melting in the lower crust. Permian metamorphism and magmatism has extensively been recorded and dated in the Central, Eastern, and Southern Alps. However, Permian metamorphic ages in the Western Alps so far are constrained by very few and sparsely distributed data. The present study fills this gap. We present U/Pb ages of metamorphic zircon from several Adria-derived continental units now situated in the Western Alps, defining a range between 286 and 266 Ma. Trace element thermometry yields temperatures of 580-890°C from Ti-in-zircon and 630-850°C from Zr-in-rutile for Permian metamorphic rims. These temperature estimates, together with preserved mineral assemblages (garnet-prismatic sillimanite-biotite-plagioclase-quartz-K-feldspar-rutile), define pervasive upper-amphibolite to granulite facies conditions for Permian metamorphism. U/Pb ages from this study are similar to Permian ages reported for the Ivrea Zone in the Southern Alps and Austroalpine units in the Central and Eastern Alps. Regional comparison across the former Adriatic and European margin reveals a complex pattern of ages reported from late Palaeozoic magmatic and metamorphic rocks (and relics thereof): two late Variscan age groups (~330 and ~300 Ma) are followed seamlessly by a broad range of Permian ages (300-250 Ma). The former are associated with late-orogenic collapse; in samples from this study these are weakly represented. Clearly, dominant is the Permian group, which is related to crustal thinning, hinting to a possible initiation of continental rifting along a passive margin