Activation of valvular interstitial cells is mediated by transforming growth factor-beta1 interactions with matrix molecules.

Strategies for the tissue-engineering of living cardiac valve replacements are limited by a lack of appropriate scaffold materials that both permit cell viability and actively contribute to the growth of functional tissues. Components of the extracellular matrix can localize and modify growth factor signals, and by doing so impart instructional stimuli for direction of cell phenotype. Fibronectin, collagen I, and heparin were explored as affinity matrices for sequestering and presenting soluble signaling molecules to control differentiation of valvular interstitial cells (VICs) to myofibroblasts. VIC differentiation is commonly characterized by expression of stress fibers containing alpha smooth muscle actin (alpha-SMA), and transforming growth factor-beta1 (TGF-beta1) is a central mediator of this transition. Both fibronectin and heparin, which are known to possess TGF-beta1 binding interactions, were found to increase VIC alpha-SMA expression (120% and 258% of expression in controls), while VICs cultured on collagen I-modified substrates had diminished alpha-SMA expression (66% of control). Heparin treatment significantly stimulated VIC production of TGF-beta1 at all concentrations tested (50 to 400 mug/ml). Heparin-modified substrates were found to alter cell morphology through increased adsorption of serum proteins, specifically TGF-beta1. In sum, heparin produced alpha-SMA-positive myofibroblasts through both the de novo production of TGF-beta1, and its localization in the pericellular environment. The addition of heparin to fibronectin-modified substrates led to a synergistic increase in VIC alpha-SMA expression, produced by the reciprocal binding of fibronectin, heparin, cell-produced TGF-beta1. The characterization of molecules, both soluble and insoluble, that control VIC activation will be important for the development of tailored 3D culture environments for tissue-engineering applications.     

Domestication as a model system for niche construction theory

Niche Construction Theory (NCT) provides a powerful conceptual framework for understanding how and why humans and target species entered into domesticatory relationships that have transformed Earth’s biota, landforms, and atmosphere, and shaped the trajectory of human cultural development. NCT provides fresh perspective on how niche-constructing behaviors of humans and plants and animals promote co-evolutionary interactions that alter selection pressures and foster genetic responses in domesticates. It illuminates the role of niche-altering activities in bequeathing an ecological inheritance that perpetuates the co-evolutionary relationships leading to domestication, especially as it pertains to traditional ecological knowledge and the transmission of learned behaviors aimed at enhancing returns from local environments. NCT also provides insights into the contexts and mechanisms that promote cooperative interactions in both humans and target species needed to sustain niche-constructing activities, ensuring that these activities produce an ecological inheritance in which domesticates play an increasing role. A NCT perspective contributes to on-going debates in the social sciences over explanatory frameworks for domestication, in particular as they pertain to issues of reciprocal causation, co-evolution, and the role of human intentionality. Reciprocally, domestication provides a model system for evaluating on-going debates in evolutionary biology concerning the impact of niche construction, phenotypic plasticity, extra-genetic inheritance, and developmental bias in shaping the direction and tempo of evolutionary change.     

HuR binds to a single site on the C/EBPbeta mRNA of 3T3-L1 adipocytes

HuR is a ligand for nuclear mRNAs containing adenylate-uridylate rich elements in the 3'-untranslated region. Once bound to the mRNA, HuR is recognized by adapter proteins which then facilitate nuclear export of the complex. In the cytosol HuR is thought to function to control stability and translation of its ligand message. In the 3T3-L1 cells HuR is constitutively expressed and localized predominantly to the nucleus in the preadipocytes. However within 30 min of exposure to the differentiation stimulus, the HuR content in the cytosol increases consistent with HuR regulating the availability of relevant mRNAs for translation. Using in vitro RNA gel shifts, we have demonstrated that the C/EBPbeta message is a ligand for HuR and that the single binding site is an adenylate-uridylate rich element in the 3'-untranslated region.     

Isolation of dsRNA-associated VLPs from the strain Cryptococcus hungaricus CBS 6569

Double-stranded RNAs (dsRNAs) with molecular masses 1.7 and 5.0 kbp, respectively, were isolated from the strain Cryptococcus hungaricus CBS 6569. The dsRNAs were copurified with eicosahedric virus-like particles, 29 nm in diameter. This strain produced a protease-sensitive `toxin' which inhibited the growth of strain C. hungaricus CBS 4214. The toxin had maximum activity at pH 3.7. The highest toxin amount was attained after a culture period of four days.     

In vivo behavior of copper-64-labeled methanephosphonate tetraaza macrocyclic ligands

Copper-64 ( T(1/2)=12.7 h; beta(+): 0.653 MeV, 17.4%; beta(-): 0.578 MeV, 39%) is produced in a biomedical cyclotron and has applications in both imaging and therapy. Macrocyclic chelators are widely used as bifunctional chelators to bind copper radionuclides to antibodies and peptides owing to their relatively high kinetic stability. In this paper, we evaluated three tetraaza macrocyclic ligands with two, three, and four pendant methanephosphonate functional groups. DO2P [1,4,7,10-tetraazacyclododecane-1,7-di(methanephosphonic acid)], DO3P [1,4,7,10-tetraazacyclododecane-1,4,7-tri(methanephosphonic acid)], and DOTP [1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(methanephosphonic acid)] were all radiolabeled with (64)Cu in high radiochemical yields. Copper-64-labeled DO2P and DOTP were highly stable in rat serum out to 24 h, while (64)Cu-DO3P remained 73% intact, with the remainder possibly forming a (64)Cu(.)2DO3P dimer by 24 h. The biodistribution experiments were performed in normal Sprague-Dawley rats. Of the three complexes, (64)Cu-DO2P demonstrated the most optimal clearance through the blood and liver. Copper-64-DO3P and (64)Cu-DOTP exhibited higher liver uptake and longer retention of liver activity, possibly because of the large negative charge of the complexes under physiological conditions. All three (64)Cu-labeled complexes showed high accumulation in bone, likely due to the binding of the methanephosphonate groups to hydroxyapatite. These results suggest that this series of methanephosphonate macrocyclic ligands may be useful as potential bone-imaging agents. The thermodynamic stability constants of the Cu(II) complexes with these three ligands were determined, and were found to be significantly higher than those of their acetate analogues. The Cu(II)-DO2P complex exhibited the highest stability constant among divalent transition metal ion DO2P complexes. Metabolism studies of (64)Cu-DO2P in rat liver suggest that the DO2P ligand may be used as a bifunctional chelator for copper radionuclides in radiodiagnostic or radiotherapeutic studies.     

Fire and flood: Soil‐stored seed bank and germination ecology in the endangered Carrington Falls Grevillea (Grevillea rivularis,Proteaceae)

Abstract Seed germination is dependent on the interaction between the dormancy state of a seed and the presence of favourable environmental conditions. Thus, the spectacular pulse of seedling recruitment in many Australian vegetation communities following disturbances such as fire can be attributed to changes in microsite conditions and/or the dormancy‐breaking effect of the disturbance on accumulated seed banks. Grevillea rivularis is a threatened species endemic to the area immediately above Carrington Falls in the NSW Southern Highlands. Most of the population is confined to the riparian vegetation zone in woodland and heath, and is therefore subject to periodic disturbance from fire and flood. For this species, a pulse of seedling recruitment has been recorded after fire, flood and mechanical soil disturbance. The aims of this study were to examine the density and vertical distribution of the soil‐stored seed bank and to investigate the role of heat and scarification as cues for germination of fresh and soil‐stored seed. There was a large seed bank under the canopies of established individuals (194 ± 73 seeds m^?2) and most seeds were found in the 0–2 cm and leaf‐litter layers of the soil profile. The germination response of soil‐stored and fresh seed was examined using a hierarchical series of laboratory experiments. Seeds of G. rivularis showed marked dormancy polymorphism. Thirty‐six percent of soil‐stored seed germinated without treatment, whereas no untreated fresh seeds germinated. Scarification or heating caused significant germination of dormant soil‐stored seed, but only scarification resulted in germination of dormant fresh seeds. These results highlight important differences in the dormancy state of soil‐stored and fresh seed. Thus, being a riparian species in a fire‐prone environment, the dormancy mechanisms in seeds of G. rivularis suit this species to disturbance by both fire and flood.     

Modeling Cesium Partitioning In the Rhizosphere: A Focus on the Role of Root Exudates

A conceptual model is being developed for the prediction of cesium partitioning between bound, aqueous, and phytoextracted phases in the rhizosphere. The model categorizes the processes that impact cesium partitioning into six submodels: geochemistry, physical factors, root density, microorganisms, nutrients, and root exudates. A seventh submodel (Cs fate) describes Cs movement between the three phases. Functional relationships and parametric values within and between the submodels are developed based on literature, field characterization, and laboratory experiments. Sensitivity analyses were first conducted to evaluate the effects of root exudates on Cs partitioning. The model was also used to test the sensitivity of root density, microbial population, potassium requirement and concentration, and moisture content on the concentration of root exudates and consequently on Cs partitioning. An increase in the concentration of root exudates results in a greater decrease in the bound concentration and greater increase in the aqueous and phytoextracted concentrations. In general, the other parameters affect Cs partitioning according to how they affect the fate of root exudates. However, the comprehensive nature of the system renders a complex overall effect on Cs partitioning. In summary, the model provides a framework for better understanding the complex interaction of processes that control Cs fate in the rhizosphere.