Regulating in vivo calcification of alginate microbeads

Alginate calcification has been previously reported clinically and during animal implantation; however no study has investigated the mechanism, extensively characterized the mineral, or evaluated multiple methods to regulate or eliminate mineralization. In the present study, alginate calcification was first studied in vitro: calcium-crosslinked alginate beads sequestered surrounding phosphate while forming traces of hydroxyapatite. Calcification in vivo was then examined in nude mice using alginate microbeads with and without adipose stem cells (ASCs). Variables included the delivery method, site of delivery, sex of the animal, time in vivo, crosslinking solution, and method of storage prior to delivery. Calciumcrosslinked alginate microbeads mineralized when injected subcutaneously or implanted intramuscularly after 1e6 months. More extensive analysis with histology, microCT, FTIR, XRD, and EDS showed calcium phosphate deposits throughout the microbeads with surface mineralization that closely matched hydroxyapatite found in bone. Incorporating 25 mM bisphosphonate reduced alginate calcification whereas using barium chloride eliminated mineralization. Buffering the crosslinking solution with HEPES at pH 7.3 while washing and storing samples in basal media prior to implantation also eliminated calcification in vivo. This study shows that alginate processing prior to implantation can significantly influence bulk hydroxyapatite formation and presents a method to regulate alginate calcificationAlginate calcification has been previously reported clinically and during animal implantation; however no study has investigated the mechanism, extensively characterized the mineral, or evaluated multiple methods to regulate or eliminate mineralization. In the present study, alginate calcification was first studied in vitro: calcium-crosslinked alginate beads sequestered surrounding phosphate while forming traces of hydroxyapatite. Calcification in vivo was then examined in nude mice using alginate microbeads with and without adipose stem cells (ASCs). Variables included the delivery method, site of delivery, sex of the animal, time in vivo, crosslinking solution, and method of storage prior to delivery. Calciumcrosslinked alginate microbeads mineralized when injected subcutaneously or implanted intramuscularly after 1e6 months. More extensive analysis with histology, microCT, FTIR, XRD, and EDS showed calcium phosphate deposits throughout the microbeads with surface mineralization that closely matched hydroxyapatite found in bone. Incorporating 25 mM bisphosphonate reduced alginate calcification whereas using barium chloride eliminated mineralization. Buffering the crosslinking solution with HEPES at pH 7.3 while washing and storing samples in basal media prior to implantation also eliminated calcification in vivo. This study shows that alginate processing prior to implantation can significantly influence bulk hydroxyapatite formation and presents a method to regulate alginate calcificatio

Metamorphosis of plasma turbulence-shear flow dynamics through a transcritical bifurcation

The structural properties of an economical model for a confined plasma turbulence governor are investigated through bifurcation and stability analyses. A close relationship is demonstrated between the underlying bifurcation framework of the model and typical behavior associated with low- to high-confinement transitions such as shear flow stabilization of turbulence and oscillatory collective action. In particular, the analysis evinces two types of discontinuous transition that are qualitatively distinct. One involves classical hysteresis, governed by viscous dissipation. The other is intrinsically oscillatory and non-hysteretic, and thus provides a model for the so-called dithering transitions that are frequently observed. This metamorphosis, or transformation, of the system dynamics is an important late side-effect of symmetry-breaking, which manifests as an unusual non-symmetric transcritical bifurcation induced by a significant shear flow drive.Comment: 17 pages, revtex text, 9 figures comprised of 16 postscript files. Submitted to Phys. Rev.

Behavioral Adaptations of Female Mice on the International Space Station

Adult female mice were sent to the International Space Station (ISS) as part of an early life science mission utilizing NASA's Rodent Habitat. Its primary purpose was to provide further insight into the influence of a microgravity environment on various aspects of mammalian physiology and well-being as part of an ongoing program of research aimed ultimately at understanding and ameliorating the deleterious influences of space on the human body. The present study took advantage of video collected from fixed, in-flight cameras within the habitat itself, to assess behavioral adaptations observed among in-flight mice aboard the ISS and differences in behavior with respect to a control group on the ground. Data collection consisted of several behavioral measures recorded by a trained observer with the assistance of interactive behavior analysis software. Specific behavioral measures included frequencies of conspecific interactionsociability, time spent feeding and conducting hygienic behavior, and relative durations of thigmotactic behavior, which is commonly used as an index of anxiety. Data were used to test tentative hypotheses that such behaviors differ significantly across mice under microgravity versus 1g conditions, and the assumption that the novel experience of microgravity itself may represent an initially anxiogenic stimulus which an animal will eventually acclimate to, perhaps through habituation

A PDZ-Binding Motif is Essential but Not Sufficient to Localize the C Terminus of CFTR to the Apical Membrane

Localization of ion channels and transporters to the correct membrane of polarized epithelia is important for vectorial ion movement. Prior studies have shown that the cytoplasmic carboxyl terminus of the cystic fibrosis transmembrane conductance regulator (CFTR) is involved in the apical localization of this protein. Here we show that the C-terminal tail alone, or when fused to the green fluorescent protein (GFP), can localize to the apical plasma membrane, despite the absence of transmembrane domains. Co-expression of the C terminus with full-length CFTR results in redistribution of CFTR from apical to basolateral membranes, indicating that both proteins interact with the same target at the apical membrane. Amino acid substitution and deletion analysis confirms the importance of a PDZ-binding motif D-T-R-L\u3e for apical localization. However, two other C-terminal regions, encompassing amino acids 1370-1394 and 1404-1425 of human CFTR, are also required for localizing to the apical plasma membrane. Based on these results, we propose a model of polarized distribution of CFTR, which includes a mechanism of selective retention of this protein in the apical plasma membrane and stresses the requirement for other C-terminal sequences in addition to a PDZ-binding motif

The hydrostatic-to-lensing mass bias from resolved X-ray and optical-IR data

An accurate reconstruction of galaxy cluster masses is key to use this population of objects as a cosmological probe. In this work we present a study on the hydrostatic-to-lensing mass scaling relation for a sample of 53 clusters whose masses were reconstructed homogeneously in a redshift range between z = 0.05 and 1.07. The M500 mass for each cluster was indeed inferred from the mass profiles extracted from the X-ray and lensing data, without using a priori observable-mass scaling relations. We assessed the systematic dispersion of the masses estimated with our reference analyses with respect to other published mass estimates. Accounting for this systematic scatter does not change our main results, but enables the propagation of the uncertainties related to the mass reconstruction method or used dataset. Our analysis gives a hydrostatic-to-lensing mass bias of (1 − b) = 0.739+−00075070 and no evidence of evolution with redshift. These results are robust against possible subsample differences

The hydrostatic-to-lensing mass bias from resolved X-ray and optical-IR data

An accurate reconstruction of galaxy cluster masses is key to use this population of objects as a cosmological probe. In this work we present a study on the hydrostatic-to-lensing mass scaling relation for a sample of 53 clusters whose masses were reconstructed homogeneously in a redshift range between $z= 0.05$ and $1.07$. The $M_{500}$ mass for each cluster was indeed inferred from the mass profiles extracted from the X-ray and lensing data, without using a priori observable-mass scaling relations. We assessed the systematic dispersion of the masses estimated with our reference analyses with respect to other published mass estimates. Accounting for this systematic scatter does not change our main results, but enables the propagation of the uncertainties related to the mass reconstruction method or used dataset. Our analysis gives a hydrostatic-to-lensing mass bias of $(1-b) =0.739^{+0.075}_{-0.070}$ and no evidence of evolution with redshift. These results are robust against possible subsample differences