The success of polyethyleneimine (PEI) as a non-viral based gene delivery vector has been attributed to its proton buffering capacity. Despite the great interest in PEI for its use in non-viral based gene delivery, the protonation behavior of PEI in solution is not well understood. Earlier experimental studies have reported inconsistent values of the protonation state of PEI. In this work, we report our investigation of the protonation behavior of a realistic linear PEI (lPEI) with computational approaches. Reported experimental pKa values of several diamine compounds are first examined. A screened coulombic interaction with a distance dependence dielectric is shown to reproduce the shifted pKa values of the model diamine compounds. Then atomistic molecular dynamics simulations of lPEI chain with 20 repeating units are performed and the results are used to provide parameters for a coarse-grained polyamine model. The screened coulombic interaction is then incorporated in the coarse-grained lPEI chain and computational titrations are performed. The obtained computational titration curves of lPEI in solutions were found to be in best agreement with experimental results by Smits et al, but the computational titration curves have too strong of a dependence on salt concentration compared to the experimental results by Smits et al. Disregarding the discrepancy in the salt dependence, our computational titrations reveal that approximately 55% of the lPEI amine groups are protonated under physiological conditions in solution with a nearly alternating arrangement of protonated and non-protonated amines. Titrations of lPEI in the presence of a polyanion are also performed to determine how the charge state of lPEI could be affected by complexation with DNA in gene therapy preparations. While the presence of the polyanion increases the degree of protonation of the PEI, many of PEI amines remain unprotonated under physiological conditions, providing evidence that PEI complexed with DNA could still have proton buffering capacity. Potential sources of error that have resulted in the inconsistency of previously reported protonation states of PEI were also discussed.
The dynamics of the exchange of chains between micelles formed by diblock copolymers in dilute solution in a selective solvent has been studied using fluorescence measurements. The samples are polystyrene-6Zoc*-poly(oxyethylene) with a single fluorescent label (either naphthalene or pyrene) covalently attached at the junction between the blocks. The critical micelle concentration (cmc) of each sample can be determined from the concentration dependence of the integrated emission intensity, provided the cmc is high enough to permit detection. In order to study the kinetics, micelles of the two differently labeled samples were first prepared at the same concentration, solvent, and temperature, but in two different containers. The contents of the containers were then mixed, and the efficiency of nonradiative singlet energy transfer from naphthalene to pyrene was measured as a function of time. The time dependence of the intensity of the emission from naphthalene can be fitted to a sum of two exponentials, with time constants that differ by at least an order of magnitude. Activation energies are somewhat smaller for the faster process than for the slower process, but in both cases they are on the order of 102 kJ mol-1 under the conditions where they can be measured. We have not been able to account for this result with a kinetic scheme that assumes the exchange of chains between the micelles takes place exclusively via the population of free chains. This difficulty suggests that an additional mechanism for the exchange of chains may be active.
The authors analyzed extensively the dynamics of polymer chains in solutions simulated with dissipative particle dynamics (DPD), with a special focus on the potential influence of a low Schmidt number of a typical DPD fluid on the simulated polymer dynamics. It has been argued that a low Schmidt number in a DPD fluid can lead to underdevelopment of the hydrodynamic interaction in polymer solutions. The authors' analyses reveal that equilibrium polymer dynamics in dilute solution, under typical DPD simulation conditions, obey the Zimm [J. Chem. Phys. 24, 269 (1956)] model very well. With a further reduction in the Schmidt number, a deviation from the Zimm model to the Rouse model is observed. This implies that the hydrodynamic interaction between monomers is reasonably developed under typical conditions of a DPD simulation. Only when the Schmidt number is further reduced, the hydrodynamic interaction within the chains becomes underdeveloped. The screening of the hydrodynamic interaction and the excluded volume interaction as the polymer volume fraction is increased are well reproduced by the DPD simulations. The use of soft interaction between polymer beads and a low Schmidt number do not produce noticeable problems for the simulated dynamics at high concentrations, except for the entanglement effect which is not captured in the simulations.
We showed that IGF-I deficiency impaired osteoclastogenesis directly and/or indirectly by altering the interaction between stromal/osteoblastic cells and osteoclast precursors, reducing RANKL and M-CSF production. These changes lead to impaired bone resorption, resulting in high BV/TV in IGF-I null mice.
We showed that the IGF-IR-null mutation in mature osteoblasts leads to less bone and decreased periosteal bone formation and impaired the stimulatory effects of PTH on osteoprogenitor cell proliferation and differentiation.Introduction: This study was carried out to examine the role of IGF-I signaling in mediating the actions of PTH on bone. Materials and Methods: Three-month-old mice with an osteoblast-specific IGF-I receptor null mutation (IGF-IR OBKO) and their normal littermates were treated with vehicle or PTH (80 g/kg body weight/d for 2 wk). Structural measurements of the proximal and midshaft of the tibia were made by CT. Trabecular and cortical bone formation was measured by bone histomorphometry. Bone marrow stromal cells (BMSCs) were obtained to assess the effects of PTH on osteoprogenitor number and differentiation. Results: The fat-free weight of bone normalized to body weight (FFW/BW), bone volume (BV/TV), and cortical thickness (C.Th) in both proximal tibia and shaft were all less in the IGF-IR OBKO mice compared with controls. PTH decreased FFW/BW of the proximal tibia more substantially in controls than in IGF-IR OBKO mice. The increase in C.Th after PTH in the proximal tibia was comparable in both control and IGF-IR OBKO mice. Although trabecular and periosteal bone formation was markedly lower in the IGF-IR OBKO mice than in the control mice, endosteal bone formation was comparable in control and IGF-IR OBKO mice. PTH stimulated endosteal bone formation only in the control animals. Compared with BMSCs from control mice, BMSCs from IGF-IR OBKO mice showed equal alkaline phosphatase (ALP) + colonies on day 14, but fewer mineralized nodules on day 28. Administration of PTH increased the number of ALP
Counterfeiting is a global epidemic that is compelling the development of new anticounterfeiting strategy. Herein, we report a novel multiple anticounterfeiting encoding strategy of invisible fluorescent quick response (QR) codes with emission color as information storage unit. The strategy requires red, green, and blue (RGB) light-emitting materials for different emission colors as encrypting information, single excitation for all of the emission for practicability, and ultraviolet (UV) excitation for invisibility under daylight. Therefore, RGB light-emitting nanoscale metal-organic frameworks (NMOFs) are designed as inks to construct the colorful light-emitting boxes for information encrypting, while three black vertex boxes were used for positioning. Full-color emissions are obtained by mixing the trichromatic NMOFs inks through inkjet printer. The encrypting information capacity is easily adjusted by the number of light-emitting boxes with the infinite emission colors. The information is decoded with specific excitation light at 275 nm, making the QR codes invisible under daylight. The composition of inks, invisibility, inkjet printing, and the abundant encrypting information all contribute to multiple anticounterfeiting. The proposed QR codes pattern holds great potential for advanced anticounterfeiting.
We showed that unloading markedly diminished the effects of IGF-I to activate its signaling pathways, and the disintegrin echistatin showed a similar block in osteoprogenitor cells. Furthermore, unloading decreased ␣V3 integrin expression. These results show that skeletal unloading induces resistance to IGF-I by inhibiting activation of the IGF-I signaling pathways at least in part through downregulation of integrin signaling. Introduction:We have previously reported that skeletal unloading induces resistance to insulin-like growth factor-I (IGF-I) with respect to bone formation. However, the underlying mechanism remains unclear. The aim of this study was to clarify how skeletal unloading induces resistance to the effects of IGF-I administration in vivo and in vitro with respect to bone formation. Materials and Methods:We first determined the response of bone to IGF-I administration in vivo during skeletal unloading. We then evaluated the response of osteoprogenitor cells isolated from unloaded bones to IGF-I treatment in vitro with respect to activation of the IGF-I signaling pathways. Finally we examined the potential role of integrins in mediating the responsiveness of osteoprogenitor cells to IGF-I. Results: IGF-I administration in vivo significantly increased proliferation of osteoblasts. Unloading markedly decreased proliferation and blocked the ability of IGF-I to increase proliferation. On a cellular level, IGF-I treatment in vitro stimulated the activation of its receptor, Ras, ERK1/2 (p44/42 MAPK), and Akt in cultured osteoprogenitor cells from normally loaded bones, but these effects were markedly diminished in cells from unloaded bones. These results were not caused by altered phosphatase activity or changes in receptor binding to IGF-I. Inhibition of the Ras/MAPK pathway was more impacted by unloading than that of Akt. The disintegrin echistatin (an antagonist of the ␣V3 integrin) blocked the ability of IGF-I to stimulate its receptor phosphorylation and osteoblast proliferation, similar to that seen in cells from unloaded bone. Furthermore, unloading significantly decreased the mRNA levels both of ␣V and 3 integrin subunits in osteoprogenitor cells. Conclusion:These results indicate that skeletal unloading induces resistance to IGF-I by inhibiting the activation of IGF-I signaling pathways, at least in part, through downregulation of integrin signaling, resulting in decreased proliferation of osteoblasts and their precursors.
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