Study on the preparation of chitosan-alginate complex membrane and the effects on adhesion and activation of endothelial cells

Biomaterials that support vascularization without activating immune competent endothelial cells are desirous in immunoisolation. The aim of the present study was to evaluate effect of chitosan-alginate membrane on adhesion and activation of endothelial cells. The AFM photographs showed that the CS/ALG membrane surface roughness values were less than CS membrane and ALG membrane. The contact angle of CS/ALG membrane was between ALG membrane and CS membrane. The surface roughness and hydrophilicity of CS/ALG membrane affect the adhesion and activation on endothelial cells. The CS/ALG membrane did not allow the majority of cells to adhere well but maintained their viability. The membrane leach-outs were nontoxic to the cells, as confirmed by tetrazolium reduction (MTT) and trypan blue assays. The growth curves of the endothelial cells desquamated from the CS/ALG membrane suggested that the endothelial cells maintain their activation. Taken together these results point out that CS/ALG membrane is compatible with endothelial cells and maintains their nonactivated status.     

A collagen based vitro model of angiogenesis designed for tissue-engineering material

Angiogenesis is central importance to tissue-engineering. Many vitro models are developed to study the mechanism of angiogenesis, making a great deal of contribution to drug development against tumor, and often may be expensive, time-consuming. Till now, few reported models have been applied to evaluating the effect of degradation fluid of tissue-engineering material to angiogenesis. In present study, we used ECV304 cell as the model cell line, type I collagen matrix that contained no stimulatory factors as a culture substratum to develop a testing model. Tube-like structure (TLS) formed within 8 h on lower density of collagen (0.2, 0.5 mg/ml), which is not found on dense collagen (1, 2 mg/ml). After ECV304 cells were seeded on the surface of collagen matrix, adherence occurred within 1 h. Soon afterwards, ECV304 cells migrated into cell aggregates, then sent out elongated cell processes to form TLS by cytoplasmic anastomosis. Proliferation was obviously perceived during the course. To investigate the efficiency of the model, we took poly(lactic acid) (PLA) degradation fluid with degradation time varying from 1 to 120 days as the testing material. TLS formation is enhanced by ECV304 cells exposed to early degradation fluid before 50-day point, and the trend of inhibition grew as the degradation time increased. Further, no formation was found in degradation fluid after 90-day point. The model is sensitive to the surrounding environment, and can demonstrate the effects of testing material quantitatively to angiogenesis. In summary, the simplicity, reproducibility and miniaturized character of the model described here may make it highly useful as a medium to test the effect of degradation fluid of tissue-engineering material to angiogenesis.     

Bilayers of neutral lipids bear a small but significant charge

Many experiments done on neutral lipid bilayers in pure water show weak repulsions. These weak forces prevent vesicles from adhering and are generally overcome by adding some salt in the aqueous medium. They also appear as stray repulsions in surface forces measurements made on lipid bilayers. Using a surface forces apparatus in pure water and in salt solution, we have measured the forces between two stearoyl-oleoyl-phosphatidyl-choline (SOPC) bilayers and between two dimiristoyl-phosphatidyl-ethanolamine (DMPE) bilayers. The results show that the repulsions are due to a small amount of negative charges coming from impurities in SOPC. This was quantitatively confirmed by electrophoretic measurements. There are 3 times less charges in the case of DMPE layers. The effect of these charges which is negligible at high salt concentration may significantly affect the adhesion energy and behaviour of neutral lipid bilayers between 0 and salt. Received 18 December 1998     

On the construction of complex networks with optimal Tsallis entropy

In this work, we first formulate the Tsallis entropy in the context of complex networks. We then propose a network construction whose topology maximizes the Tsallis entropy. The growing network model has two main ingredients: copy process and random attachment mechanism (C-R model). We show that the resulting degree distribution exactly agrees with the required degree distribution that maximizes the Tsallis entropy. We also provide another example of network model using a combination of preferential and random attachment mechanisms (P-R model) and compare it with the distribution of the Tsallis entropy. In this case, we show that by adequately identifying the exponent factor q, the degree distribution can also be written in the q-exponential form. Taken together, our findings suggest that both mechanisms, copy process and preferential attachment, play a key role for the realization of networks with maximum Tsallis entropy. Finally, we discuss the interpretation of q parameter of the Tsallis entropy in the context of complex networks.     

Role of solvent polarity on the fluorescence quenching of newly synthesized 7,8-benzo-4-azidomethyl coumarin by aniline in benzene-acetonitrile mixtures

The fluorescence quenching of 7,8-benzo-4-azidomethyl coumarin by aniline has been studied in different solvent mixtures of benzene (BN) and acetonitrile (AN) at room temperature. The quenching is found to be appreciable and a positive deviation from linearity was observed in the Stern-Volmer (S-V) plot in all the solvent mixtures. Various parameters for the quenching process have been determined by sphere of action static quenching model and finite sink approximation model. The magnitudes of these rate parameters indicate that positive deviation in the S-V plot is both due to static and dynamic processes.     

Comparative analysis of the gene co-regulatory network of normal and cancerous lung

The gene co-regulatory networks of normal and cancerous lung cells are constructed based on microarrays. In these networks, nodes represent genes and links represent co-regulatory relationships between genes. A comparison of these two networks indicates that they are significantly different in terms of both global statistical features and also in some details. The results imply at least four interesting messages: First, lung cancer is not simply a disordered system since it also has some rules in itself. Second, both of these networks can be decomposed into several functional modules, each of which is responsible for certain biological function. Third, among those functional modules, the module which is responsible for immune reactions seems to work independently rather than co-operating with others. Fourth, the phenomenon of module dysfunction is observed in cancerous lung cells.     

Speed of adaptation in structured populations

Adaptation of populations takes place with the occurrence and subsequent fixation of mutations that confer some selective advantage to the individuals which acquire it. For this reason, the study of the process of fixation of advantageous mutations has a long history in the population genetics literature. Particularly, the previous investigations aimed to find out the main evolutionary forces affecting the strength of natural selection in the populations. In the current work, we investigate the dynamics of fixation of beneficial mutations in a subdivided population. The subpopulations (demes) can exchange migrants among their neighbors, in a migration network which is assumed to have either a random graph or a scale-free topology. We have observed that the migration rate drastically affects the dynamics of mutation fixation, despite of the fact that the probability of fixation is invariant on the migration rate, accordingly to Maruyama's conjecture. In addition, we have noticed a topological dependence of the adaptive evolution of the population when clonal interference becomes effective.     

Translational order in liquid-expanded lipid monolayers functionalized with nucleosides

The monolayer behavior of a lipid containing two unsaturated alkyl chains and a nucleoside derivative as polar headgroup has been investigated by the Langmuir technique. From the surface pressure vs. molecular area isotherm, the monolayer appears as a pure liquid-expanded phase and should be then considered as a two-dimensional liquid. However, grazing incidence X-ray diffraction experiments evidence a translational order that does not exist when the lipid headgroup is a choline moiety. Since unsaturated chains are expected to induce a fluid state of the monolayer at the temperature considered, this order is likely to originate from the natural tendency nucleosides have to establish among themselves -stacking interactions between the bases. The collected X-ray data are consistent with the geometrical requirements for bases stacking. Received: 3 August 1998 / Revised: 16 September 1998 / Accepted: 21 September 1998     

Active membranes studied by X-ray scattering

In view of recent theories of “active” membranes, we have studied multilamellar phospholipid membrane stacks with reconstituted transmembrane protein bacteriorhodopsin (BR) under different illumination conditions by X-ray scattering. The light-active protein is considered as an active constituent which drives the system out of equilibrium and is predicted to change the collective fluctuation properties of the membranes. Using X-ray reflectivity, X-ray non-specular (diffuse) scattering, and grazing incidence scattering, we find no detectable change in the scattering curves when changing the illumination condition. In particular the intermembrane spacing d remains constant, after eliminating hydration-related artifacts by design of a suitable sample environment. The absence of any observable non-equilibrium effects in the experimental window is discussed in view of the relevant parameters and recent theories.     

Structurally constrained protein evolution: results from a lattice simulation

We simulate the evolution of a protein-like sequence subject to point mutations, imposing conservation of the ground state, thermodynamic stability and fast folding. Our model is aimed at describing neutral evolution of natural proteins. We use a cubic lattice model of the protein structure and test the neutrality conditions by extensive Monte Carlo simulations. We observe that sequence space is traversed by neutral networks, i.e. sets of sequences with the same fold connected by point mutations. Typical pairs of sequences on a neutral network are nearly as different as randomly chosen sequences. The fraction of neutral neighbors has strong sequence to sequence variations, which influence the rate of neutral evolution. In this paper we study the thermodynamic stability of different protein sequences. We relate the high variability of the fraction of neutral mutations to the complex energy landscape within a neutral network, arguing that valleys in this landscape are associated to high values of the neutral mutation rate. We find that when a point mutation produces a sequence with a new ground state, this is likely to have a low stability. Thus we tentatively conjecture that neutral networks of different structures are typically well separated in sequence space. This result indicates that changing significantly a protein structure through a biologically acceptable chain of point mutations is a rare, although possible, event. Received 8 July 1999     

Folding kinetics of two-state proteins based on the model of general random walk in native contact number space

We investigated the folding kinetics of a series of two-state proteins by using the model of general random walk in native contact number space, and derive the observed linear relationship between the logarithms of the folding rate constants and the numbers of native contacts from “kinetic viewpoint”. The protein folding speed limit and stability in this model are consistent with experimental observations.     

Percolation in a network with long-range connections: Implications for cytoskeletal structure and function

Cell shape, signaling, and integrity depend on cytoskeletal organization. In this study we describe the cytoskeleton as a simple network of filamentary proteins (links) anchored by complex protein structures (nodes). The structure of this network is regulated by a distance-dependent probability of link formation as P=p/d^s, where p regulates the network density and s controls how fast the probability for link formation decays with node distance (d). It was previously shown that the regulation of the link lengths is crucial for the mechanical behavior of the cells. Here we examined the ability of the two-dimensional network to percolate (i.e. to have end-to-end connectivity), and found that the percolation threshold depends strongly on s. The system undergoes a transition around s=2. The percolation threshold of networks with s<2 decreases with increasing system size L, while the percolation threshold for networks with s>2 converges to a finite value. We speculate that s<2 may represent a condition in which cells can accommodate deformation while still preserving their mechanical integrity. Additionally, we measured the length distribution of F-actin filaments from publicly available images of a variety of cell types. In agreement with model predictions, cells originating from more deformable tissues show longer F-actin cytoskeletal filaments.     

Active Control of Cellular Orientation through In-Situ Stress in the Substrate

We investigate the orientation of cells on substrates to find possible methods for controlling the cellular orientation. The force dipole model is employed in our modelling and simulation. The elastic interaction between cells as well as the elastic interaction between the cell and in-situ stress field in the substrate are found to be the two main physical mechanisms to control the cellular orientation. The former interaction dominates the cellular orientation when the in-situ stress is small, while the later dominates when the in-situ stress is large enough. Two cells tend to align perpendicularly on a free substrate, but the cellular orientation varies with the increasing in-situ stress. Two cells tendto align in parallel when the normal stress is large enough. Their direction is perpendicular to the extension stress direction or parallel to the compression stress direction. When the positive in-situ shear stress is large enough, the two cells tend to align at -45°. Based on this theoretical simulation, it is believed that the cellular orientation on substrates can be controlled by thein-situ stresses.     

A compounded rare-earth iron garnet single crystal exhibiting stable Faraday rotation against wavelength and temperature variation in the 1.55 μm band

The Bi, Tb and Yb partially substituted iron garnet bulk single crystals of Tb_3−x−yYb_yBi_xFe₅O₁₂ were grown by using Bi₂O₃/B₂O₃ as flux and accelerated crucible rotation technique for single-crystal growth. Faraday rotation (FR) spectra showed that the specific FR of the (Tb_0.91Yb_1.38Bi_0.71)Fe₅O₁₂ crystal under magnetic field at saturation was measured to be about −1617°/cm at λ=1.55 μm, Faraday rotation wavelength coefficient (FWC, 0.009%/nm) in the wavelength range of 1.50–1.62 μm and Faraday rotation temperature coefficient (FTC, 3.92×10⁻⁵/K) at λ=1.55 μm were even smaller than that of YIG. It is proven that through combining two types of Bi-substituted rare-earth iron garnets with opposite FWC and FTC signs, the compound rare-earth iron garnets with low FWC and FTC may be obtained due to the compensation effect. The saturation magnetization of (Tb_0.91Yb_1.38Bi_0.71) Fe₅O₁₂ crystal is 0.48×10⁶ A/M and is also much smaller than that of YIG. We have found empirically that there is a simple relationship between the FR θ_f(x) and Bi content x for Tb_3−x−yYb_yBi_xFe₅O₁₂, which is given by θ_f(x)=(−2759x+400)°/cm.     

Using 3D fluid-structure interaction model to analyse the biomechanical properties of erythrocyte

This Letter presents a newly developed three-dimensional fluid-structure interaction model of the red blood cell (RBC). The model consists of a deformable liquid capsule modelled as Newtonian fluid enclosed by a hyperelastic membrane with viscoelastic property. Numerical results show that viscosity in the cytoplasm affects the deformed shape of RBC under loading. This observation is contrary to the earlier belief that viscosity of the cytoplasm can be neglected. Numerical simulations carried out to investigate large deformation induced on the RBC model using direct tensile forces show significant improvement in terms of correlation with experimental results. The membrane shear modulus estimated from the model ranges between 3.7 to compares well with results obtained from micropipette aspiration experiments.     

A study of Barstar folding events using boundary value simulations

From extensive biophysical studies of protein folding, two competing mechanisms emerged: hydrophobic collapse and the framework model. Our protein of choice is Barstar—a barnase inhibitor. The approximation algorithm we used to study Barstar folding trajectories is called SDEL—stochastic difference equation in length. Using the native structure as the final boundary value and a collection of unfolded structures as the varying initial boundary value, SDEL calculates an ensemble of least action pathways between these boundaries. The results are atomically detailed folding pathways, with as many intermediate structures as you request in the input. We generated 12 pathways, starting from a structurally wide selection of unfolded conformations. Using the protein's radius of gyration as our primary reaction coordinate, we tracked H-bonds, dihedral angles, native and non-native contacts, and energy along the folding pathways. This paper will follow our findings, with special emphasis on pinpointing hydrophobic collapse as a more appropriate mechanism for Barstar. Comparison with pathway predictions for Barstar using experimental techniques will also be discussed.     

Influence of surface treatments on the shear bond strength of orthodontic brackets to porcelain

The purpose of this study was to investigate the effect of various surface treatments after different storage time and thermocycling on the shear bond strength of orthodontic brackets to the feldspathic porcelain surfaces. 128 disc-shaped porcelain specimens were randomly assigned to the following surface treatments: 9.6% HFA, 9.6% HFA combined with silane, 50 μ aluminum trioxide sandblasting followed by silane and application of silane after 37% phosphoric acid. Metal or ceramic brackets were bonded onto each treated porcelain facet with light cured resin. The samples were stored in 37 °C water 1 day or 7 days, thermocycled 500 times from 5 to 55 °C. The shear bond strengths were measured (1 mm/min), and statistically analyzed. The bond failure sites were classified according to ARI system. The surface of the glazed, sandblasted, hydrofluoric and phosphoric acid etched porcelain were examined with SEM. All groups achieved reasonable bond strengths to withstand the application of orthodontic forces. Water storage for 7 days caused lower shear bond strength than that of 1 day. But there is no statistically significant difference between the two groups. The mean shear bond strength provided by ceramic bracket with mechanical retention had no statistical difference with that of metal bracket. Therefore, the optimal treatment for orthodontic brackets bonding to feldspathic porcelain was to apply phosphoric acid combined with silane.     

Evolution of canalizing Boolean networks

Boolean networks with canalizing functions are used to model gene regulatory networks. In order to learn how such networks may behave under evolutionary forces, we simulate the evolution of a single Boolean network by means of an adaptive walk, which allows us to explore the fitness landscape. Mutations change the connections and the functions of the nodes. Our fitness criterion is the robustness of the dynamical attractors against small perturbations. We find that with this fitness criterion the global maximum is always reached and that there is a huge neutral space of 100% fitness. Furthermore, in spite of having such a high degree of robustness, the evolved networks still share many features with “chaotic” networks.