Characterization of cell viability during bioprinting processes

Bioprinting is an emerging technology in the field of tissue engineering and regenerative medicine. The process consists of simultaneous deposition of cells, biomaterial and/or growth factors under pressure through a micro-scale nozzle. Cell viability can be controlled by varying the parameters like pressure and nozzle diameter. The process itself can be a very useful tool for evaluating an in vitro cell injury model. It is essential to understand the cell responses to process-induced mechanical disturbances because they alter cell morphology and function. We carried out analysis and quantification of the degree of cell injury induced by bioprinting process. A parametric study with different process parameters was conducted to analyze and quantify cell injury as well as to optimize the parameters for printing viable cells. A phenomenological model was developed correlating the percentage of live, apoptotic and necrotic cells to the process parameters. This study incorporates an analytical formulation to predict the cell viability through the system as a function of the maximum shear stress in the system. The study shows that dispensing pressure has a more significant effect on cell viability than the nozzle diameter. The percentage of live cells is reduced significantly (by 38.75%) when constructs are printed at 40 psi compared to those printed at 5 psi.     

豚鼠腺性膀胱炎模型的建立及尿流动力学的检查

目的探讨豚鼠腺性膀胱炎动物模型的建立方法及尿流动力学检查方法。方法将28只雌性豚鼠分三组：正常对照组、生理盐水对照组和造模组。用膀胱内灌注大肠杆菌的方法制作腺性膀胱炎动物模型,7周后行尿流动力学检查及病理检测。结果尿流动力学检查发现造模组豚鼠储尿期逼尿肌不稳定发生率较正常对照组及生理盐水对照组显著增多（P〈0.001）;正常对照组无腺性膀胱炎病变,生理盐水对照组出现腺性膀胱炎1例,造模组出现7例,造模组与另外两组相比差异有统计学意义（P〈0.05）,正常对照组与生理盐水对照组相比差异无统计学意义（P〉0.05）。结论膀胱内灌注大肠杆菌可导致腺性膀胱炎,这证实了细菌感染是腺性膀胱炎的病因之一,同时为临床上的抗感染治疗提供了理论依据。此方法可用于建立腺性膀胱炎的动物模型。同时确立了腺性膀胱炎动物模型尿流动力学检查方法。     

细胞离子在振荡电磁场作用下的受力模型分析

本文通过生物细胞模型,研究振荡电场、振荡磁场以及振荡磁场产生的感应电场对细胞离子的作用机理。模型分析结果表明,电场力和罗仑兹力对细胞膜两侧的自由离子将产生加速度,振荡离子将产生周期性电位移。该模型同时也解释了脉冲电磁场比同参教的连续场产生更多的生物效应,以及连续场在开始施加和切除时的效应最大。     

Fluidity,permeability and antioxidant behaviour of model membranes incorporated with α-tocopherol and vitamin E acetate

Magnetic resonance studies reveal a marked difference between the binding of α-tocopherol and that of the corresponding acetate (vitamin E acetate) with dipalmitoylphosphatidylcholine (DPPC) vesicles. This is reflected in differences in the phase-transition curves of the DPPC vesicles incorporated with the two compounds, as well as in the ¹³C relaxation times and line widths. A model for the incorporation of these molecules in lipid bilayers has been suggested. α-Tocopherol binds strongly with the lipids, possibly through a hydrogen bond formation between the hydroxyl group of the former and one of the oxygen atoms of the latter. The possibility of such a hydrogen bond formation is excluded in vitamin E acetate, which binds loosely through the normal hydrophobic interaction. The model for lipid-vitamin interaction explains the in vitro decomposition of H₂O₂ by α-tocopherol. α-Tocopherol in conjuction with H₂O₂ can also act as a free-radical scavenger in the lipid phase. The incorporation of α-tocopherol and vitamin E acetate in DPPC vesicles enhances the permeability of lipid bilayers for small molecules such as sodium ascorbate.     

Quantifying soil organic carbon in complex landscapes: an example of grassland undergoing encroachment of woody plants

The invasion of woody plants into grass‐dominated ecosystems has occurred worldwide during the past century with potentially significant impacts on soil organic carbon (SOC) storage, ecosystem carbon sequestration and global climate warming. To date, most studies of tree and shrub encroachment impacts on SOC have been conducted at small scales and results are equivocal. To quantify the effects of woody plant proliferation on SOC at broad spatial scales and to potentially resolve inconsistencies reported from studies conducted at fine spatial scales, information regarding spatial variability and uncertainty of SOC is essential. We used sequential indicator simulation (SIS) to quantify spatial uncertainty of SOC in a grassland undergoing shrub encroachment in the Southern Great Plains, USA. Results showed that both SOC pool size and its spatial uncertainty increased with the development of woody communities in grasslands. Higher uncertainty of SOC in new shrub‐dominated communities may be the result of their relatively recent development, their more complex above‐ and belowground architecture, stronger within‐community gradients, and a greater degree of faunal disturbance. Simulations of alternative sampling designs demonstrated the effects of spatial uncertainty on the accuracy of SOC estimates and enabled us to evaluate the efficiency of sampling strategies aimed at quantifying landscape‐scale SOC pools. An approach combining stratified random sampling with unequal point densities and transect sampling of landscape elements exhibiting strong internal gradients yielded the best estimates. Complete random sampling was less effective and required much higher sampling densities. Results provide novel insights into spatial uncertainty of SOC and its effects on estimates of carbon sequestration in terrestrial ecosystem and suggest effective protocol for the estimating of soil attributes in landscapes with complex vegetation patterns.     

Formation of heterocyclic aromatic amines in model systems

Heterocyclic aromatic amines (HAs) are mutagenic and carcinogenic substances that are formed in significant amounts during heating of meat or fish at temperatures of at least 150 degrees C. To investigate the chemistry lying behind the formation of these harmful substances model systems were established. The first aim was to identify the naturally occurring precursors, namely creatinine, amino acids and carbohydrates. Later these model systems were used to develop strategies for a reduction of the content of the heterocyclic aromatic amines and for the evaluation of the reaction mechanisms that lead to the formation of these substances. All these aspects are discussed in this review.     

Drosophila melanogaster as a model system to study long-chain fatty acid amide metabolism

Long-chain fatty acid amides are cell-signaling lipids identified in mammals and, recently, in invertebrates, as well. Many details regarding fatty acid amide metabolism remain unclear. Herein, we demonstrate that Drosophila melanogaster is an excellent model system for the study long-chain fatty acid amide metabolism as we have quantified the endogenous levels of N-acylglycines, N-acyldopamines, N-acylethanolamines, and primary fatty acid amides by LC/QTOF-MS. Growth of D. melanogaster on media supplemented with [1-¹³C]-palmitate lead to a family of ¹³C-palmitate-labeled fatty acid amides in the fly heads. The [1-¹³C]-palmitate feeding studies provide insight into the biosynthesis of the fatty acid amides.     

Biophysical studies of the interactions between 14-mer and 21-mer model amphipathic peptides and membranes: Insights on their modes of action

We have investigated the interactions between synthetic amphipathic peptides and zwitterionic model membranes. Peptides with 14 and 21 amino acids composed of leucines and phenylalanines modified by the addition of crown ethers have been synthesized. The 14-mer and 21-mer peptides both possess a helical amphipathic structure as revealed by circular dichroism. To shed light on their mechanism of membrane interaction, different complementary biophysical techniques have been used such as circular dichroism, fluorescence, membrane conductivity measurement and NMR spectroscopy. Results obtained by these different techniques show that the 14-mer peptide is a membrane perturbator that facilitate the leakage of species such as calcein and Na ions, while the 21-mer peptide acts as an ion channel. ³¹P solid-state NMR experiments on multilamellar vesicles reveal that the dynamics and/or orientation of the polar headgroups are greatly affected by the presence of the peptides. Similar results have also been obtained in mechanically oriented DLPC and DMPC bilayers where different acyl chain lengths seem to play a role in the interaction. On the other hand, ²H NMR experiments on multilamellar vesicles demonstrate that the acyl chain order is affected differently by the two peptides. Based on these studies, mechanisms of action are proposed for the 14-mer and 21-mer peptides with zwitterionic membranes.