Carbonic Anhydrase is Required for Statoconia Homeostasis in Organ Cultures of Statocysts from Aplysia californica

A novel organ culture system has been developed to study the regulation of statoconia production in the gravity sensing organ in Aplysia californica. Statocysts were cultured in Leibovitz (LI5) medium supplemented with salts and Aplysia haemolymph for four days at 17 C. The viability of the system was evaluated by examining four parameters: statocyst morphology, the activity of the mechanosensory cilia in the statocyst, production of new statoconia during culture and change in statoconia volume after culture. There were no morphological differences in statocysts before and after culture when ciliary beating was maintained. There was a 29% increase in the number of statoconia after four days in culture. Mean statocyst, statolith and statoconia volumes were not affected by culture conditions. The presence of carbonic anhydrase in the statocysts was shown using immunohistochemistry. When statocysts were cultured in the presence of 4.0 x 10(exp -4) M acetazolamide to inhibit the enzyme activity, there was a decrease in statoconia production and statoconia volume, indicating a role for this enzyme in statoconia homeostasis, potentially, via pH regulation. These studies are the first to report a novel system for the culture of statocysts and show that carbonic anhydrase is involved in the regulation of statoconia volume and production

Extracellular Matrix Vesicles in Endochondral Bone Development and in Healing After Injury

Matrix vesicles are fundamental components of initial mineral formation in endochondral and primary ossification. Regulation of their structure and function is multi-faceted and involves genomic and nongenomic pathways. Their function in mineralization may include matrix maturation and conveying cellular signals to the extracellular matrix as well. Intravesicular hydroxyapatite crystal formation is associated with primary mineralization; subsequent crystal formation, particularly at later stages, appears to be mediated by other mechanisms

Selective laser sintering of hydroxyapatite reinforced polyethylene composites for bioactive implants and tissue scaffold development

Selective laser sintering (SLS) has been investigated for the production of bioactive implants and tissue scaffolds using composites of high-density polyethylene (HDPE) reinforced with hydroxyapatite (HA) with the aim of achieving the rapid manufacturing of customized implants. Single-layer and multilayer block specimens made of HA-HDPE composites with 30 and 40 vol % HA were sintered successfully using a CO2 laser sintering system. Laser power and scanning speed had a significant effect on the sintering behaviour. The degree of particle fusion and porosity were influenced by the laser processing parameters, hence control can be attained by varying these parameters. Moreover, the SLS processing allowed exposure of HA particles on the surface of the composites and thereby should provide bioactive products. Pores existed in the SLS-fabricated composite parts and at certain processing parameters a significant fraction of the pores were within the optimal sizes for tissue regeneration. The results indicate that the SLS technique has the potential not only to fabricate HA-HDPE composite products but also to produce appropriate features for their application as bioactive implants and tissue scaffolds

Singular solutions of fully nonlinear elliptic equations and applications

We study the properties of solutions of fully nonlinear, positively homogeneous elliptic equations near boundary points of Lipschitz domains at which the solution may be singular. We show that these equations have two positive solutions in each cone of $\mathbb{R}^n$, and the solutions are unique in an appropriate sense. We introduce a new method for analyzing the behavior of solutions near certain Lipschitz boundary points, which permits us to classify isolated boundary singularities of solutions which are bounded from either above or below. We also obtain a sharp Phragm\'en-Lindel\"of result as well as a principle of positive singularities in certain Lipschitz domains.Comment: 41 pages, 2 figure

Anthropogenic Disturbance and Population Viability of Woodland Caribou in Ontario

One of the most challenging tasks in wildlife conservation and management is to clarify how spatial variation in land cover due to anthropogenic disturbance influences wildlife demography and long‐term viability. To evaluate this, we compared rates of survival and population growth by woodland caribou (Rangifer tarandus caribou) from 2 study sites in northern Ontario, Canada that differed in the degree of anthropogenic disturbance because of commercial logging and road development, resulting in differences in predation risk due to gray wolves (Canis lupus). We used an individual‐based model for population viability analysis (PVA) that incorporated adaptive patterns of caribou movement in relation to predation risk and food availability to predict stochastic variation in rates of caribou survival. Field estimates of annual survival rates for adult female caribou in the unlogged ( x̄ = 0.90) and logged ( x̄ = 0.76) study sites recorded during 2010–2014 did not differ significantly (P \u3e 0.05) from values predicted by the individual‐based PVA model (unlogged:  x̄ = 0.87; logged:  x̄ = 0.79). Outcomes from the individual‐based PVA model and a simpler stage‐structured matrix model suggest that substantial differences in adult survival largely due to wolf predation are likely to lead to long‐term decline of woodland caribou in the commercially logged landscape, whereas the unlogged landscape should be considerably more capable of sustaining caribou. Estimates of population growth rates (λ) for the 2010–2014 period differed little between the matrix model and the individual‐based PVA model for the unlogged (matrix model  x̄ = 1.01; individual‐based model x̄ = 0.98) and logged landscape (matrix model x̄ = 0.88; individual‐based model x̄ = 0.89). We applied the spatially explicit PVA model to assess the viability of woodland caribou across 14 woodland caribou ranges in Ontario. Outcomes of these simulations suggest that woodland caribou ranges that have experienced significant levels of commercial forestry activities in the past had annual growth rates 0.96. These differences were strongly related to regional variation in wolf densities. Our results suggest that increased wolf predation risk due to anthropogenic disturbance is of sufficient magnitude to cause appreciable risk of population decline in woodland caribou in Ontario. © 2020 The Authors. The Journal of Wildlife Management published by Wiley Periodicals, Inc. on behalf of The Wildlife Society

In Vitro Generation of Cartilage-Carrier-Constructs on Hydroxylapatite Ceramics with Different Surface Structures

Tissue engineering approaches for healing cartilage defects are partly limited by the inability to fix cartilage to bone during implantation. To overcome this problem, cartilage can be - already in vitro - generated on a ceramic carrier which serves as bone substitute. In this study, the influence of a hydroxylapatite carrier and its surface structure on the quality of tissue engineered cartilage was investigated. Application of the carrier reduced significantly biomechanical and biochemical properties of the generated tissue. In addition, slight changes in the quality of the formed matrix, in the adhesive strength between cartilage and biomaterial and in attachment and proliferation of a chondrocyte monolayer could be observed for commercial grade carriers, with respect to modified topographies obtained by smooth grinding/polishing. These first results demonstrated an influence of the carrier and its surface structure, but further research is needed for explaining the described effects and for optimization of cartilage-carrier-constructs

Surface modification of starch based blends using potassium permanganate-nitric acid system and its effect on the adherence and proliferation of osteoblastic-like cells

The surface modification of three starch based polymeric biomaterials, using a KMnO4/NHO3 oxidizing system, and the effect of that modification on the osteoblastic cell adhesion has been investigated. The rationale of this work is as follows—starch based polymers have been proposed for use as tissue engineering scaffolds in several publications. It is known that in biodegradable systems it is quite difficult to have both cell adhesion and proliferation. Starch based polymers have shown to perform better than poly-lactic acid based materials but there is still room for improvement. This particular work is aimed at enhancing cell adhesion and proliferation on the surface of several starch based polymer blends that are being proposed as tissue engineering scaffolds. The surface of the polymeric biomaterials was chemically modified using a KMnO4/HNO3 system. This treatment resulted in more hydrophilic surfaces, which was confirmed by contact angle measurements. The effect of the treatment on the bioactivity of the surface modified biomaterials was also studied. The bioactivity tests, performed in simulated body fluid after biomimetic coating, showed that a dense film of calcium phosphate was formed after 30 days. Finally, human osteoblast-like cells were cultured on unmodified (control) and modified materials in order to observe the effect of the presence of higher numbers of polar groups on the adhesion and proliferation of those cells. Two of the modified polymers presented changes in the adhesion behavior and a significant increase in the proliferation rate kinetics when compared to the unmodified controls.FCT (Portugal) for providing the postdoctoral grant (BPD/8491/2002)

Approximate policy iteration: A survey and some new methods

We consider the classical policy iteration method of dynamic programming (DP), where approximations and simulation are used to deal with the curse of dimensionality. We survey a number of issues: convergence and rate of convergence of approximate policy evaluation methods, singularity and susceptibility to simulation noise of policy evaluation, exploration issues, constrained and enhanced policy iteration, policy oscillation and chattering, and optimistic and distributed policy iteration. Our discussion of policy evaluation is couched in general terms and aims to unify the available methods in the light of recent research developments and to compare the two main policy evaluation approaches: projected equations and temporal differences (TD), and aggregation. In the context of these approaches, we survey two different types of simulation-based algorithms: matrix inversion methods, such as least-squares temporal difference (LSTD), and iterative methods, such as least-squares policy evaluation (LSPE) and TD (λ), and their scaled variants. We discuss a recent method, based on regression and regularization, which rectifies the unreliability of LSTD for nearly singular projected Bellman equations. An iterative version of this method belongs to the LSPE class of methods and provides the connecting link between LSTD and LSPE. Our discussion of policy improvement focuses on the role of policy oscillation and its effect on performance guarantees. We illustrate that policy evaluation when done by the projected equation/TD approach may lead to policy oscillation, but when done by aggregation it does not. This implies better error bounds and more regular performance for aggregation, at the expense of some loss of generality in cost function representation capability. Hard aggregation provides the connecting link between projected equation/TD-based and aggregation-based policy evaluation, and is characterized by favorable error bounds.National Science Foundation (U.S.) (No.ECCS-0801549)Los Alamos National Laboratory. Information Science and Technology InstituteUnited States. Air Force (No.FA9550-10-1-0412