Effects of methotrexate on the viscoelastic properties of single cells probed by atomic force microscopy

Methotrexate is a commonly used anti-cancer chemotherapy drug. Cellular mechanical properties are fundamental parameters that reflect the physiological state of a cell. However, so far the role of cellular mechanical properties in the actions of methotrexate is still unclear. In recent years, probing the behaviors of single cells with the use of atomic force microscopy (AFM) has contributed much to the field of cell biomechanics. In this work, with the use of AFM, the effects of methotrexate on the viscoelastic properties of four types of cells were quantitatively investigated. The inhibitory and cytotoxic effects of methotrexate on the proliferation of cells were observed by optical and fluorescence microscopy. AFM indenting was used to measure the changes of cellular viscoelastic properties (Young’s modulus and relaxation time) by using both conical tip and spherical tip, quantitatively showing that the stimulation of methotrexate resulted in a significant decrease of both cellular Young’s modulus and relaxation times. The morphological changes of cells induced by methotrexate were visualized by AFM imaging. The study improves our understanding of methotrexate action and offers a novel way to quantify drug actions at the single-cell level by measuring cellular viscoelastic properties, which may have potential impacts on developing label-free methods for drug evaluation.     

Biofilm coupled with UV irradiation for phenol degradation and change of its community structure

The extensive use of phenol compounds and the inability to remove these compounds during wastewater treatment have resulted in the widespread occurrence of phenols in the natural environment. Phenols have been linked to serious risks to human and environmental health. Hence, the need to develop technologies that can effectively remove phenols from wastewater and source waters is a pressing challenge. In this study, light ceramic particles were immersed in activated sludge acclimated to degrade phenol, and microorganisms were allowed to attach to the particles surface to form biofilm. Then the ceramic particles with biofilm were moved into the photolytic circulating-bed biofilm reactor made of quartz glass, which was used for the degradation of phenol by three protocols: photolysis with UV light alone (P), biodegradation alone (B), and the two mechanisms operating simultaneously (photobiodegradation, P&B). The experimental results indicated that phenol removal rate was quickest by B experiment. However, P&B experiment gave more complete mineralization of phenol than that by other protocols. During P&B experiment, the microorganisms grown on porous ceramic carrier still kept the bioactivity degrading phenol, even under UV light irradiation. However, the dominant members of the bacterial community changed dramatically after the intimately coupled photobiodegradation, according to molecular biological analysis to the biofilm. Whereas Beijerinckia sp. was the dominant strain in the inoculum, it was replaced by Thauera sp. MZ1T that played a main role on degrading phenol during P&B experiment.     

Binding and Uptake of RGD-Containing Ligands to Cellular α v β 3 Integrins

The cyclic peptide, cRGDf[N(me)]V, binds to the α _v β ₃ integrin and can disrupt binding of the integrin to its natural ligands in the extracellular matrix. In this work, the ability of a water-soluble, fluorescently labeled variant of the RGD-containing peptide (cRGDfK-488) to bind to integrins on human umbilical vascular endothelial cells (HUVEC) and subsequently undergo endocytosis was characterized. This information was compared to the binding and uptake properties of an α _v β ₃ integrin-specific monoclonal antibody, LM609X. The specificity of the RGD-containing peptide is assessed by comparison with control peptide that does not bind to the α _v β ₃ integrin, cRADfK-488. Using a high purity construct, it is shown that the RGD ligand exhibits dissociation constants in the micromolar range whereas LM609X exhibits dissociation constants in the nanomolar range. However, the RGD ligand showed greater uptake following incubation at temperatures which permit endocytosis. A 7.4-fold increase in uptake of the RGD peptide was observed following a 1 h incubation with HUVEC at 37°C (an endocytosis permissive temperature), as compared to that at 4°C (an endocytosis prohibitive temperature). In contrast, only a 1.9-fold increase in cell-associated fluorescence was observed for similar incubations with LM609X. Results from fluorescence microscopy supports the notion that the RGD peptide is rapidly endocytosed at 37°C as compared to LM609X. These results are discussed with regard to previous work indicating that RGD ligands enter cells by integrin-independent pathways. These studies provide well-controlled measures of how RGD ligands stimulate endocytosis. This may be of considerable interest for intracellular delivery of ligand-associated drugs in anti-angiogenic applications.     

基于i-Tree模型的城市森林经济效益评估

城市森林的生态服务功能对改善城市环境质量具有重要作用,如何量化这些生态服务功能是使城市森林发挥最大效益的一个重要前提.随着科学的发展,对城市森林经济效益评估的量化方法已由传统的CTLA法、Burnley法、碳税法、造林成本法等发展到模型测算法.本文介绍的i-Tree模型是由美国林务局(USDA Forest Service)于2006年开发的城市森林效益评估软件,具有许多其他模型如CITY green模型等不可替代的优势,在国际城市森林研究中已得到了广泛的应用,但国内尚未推广.本文主要阐述和评价了模型中UFORE和STRATUM两个主要模块的研究及应用进展,通过作者在国内的应用实践,讨论了该模型在中国的应用前景及存在的主要问题,为城市森林管理提供科学依据.     

基于生态效率理论和TOPSIS法的工业园区循环经济发展评价

工业园区层面的生态效率评价是生态效率研究领域有待深入探索的课题。TOPSIS方法则可能是适用于生态效率评价的一种方法,它以空间统计学为基础,借助于多目标决策问题的理想解和负理想解来进行排序,能够客观地比较和评价不同样本点综合指标的优劣。本文根据生态效率和循环经济理论,结合《综合类生态工业园区标准（试行）》,建立工业园区循环经济评价指标体系,运用TOPSIS方法对苏州高新区、苏州工业园区生态工业园和无锡新区生态工业示范园区的生态效率进行综合评价,结果表明：三者的生态效率综合排名由高到低依次为：无锡新区生态工业示范园区> 苏州工业园区生态工业园> 苏州高新区,且3园区在经济发展、资源能源利用率、物质循环利用、环境污染控制等子系统有所优劣。并针对各园区的各自存在的不足提出了相应的改进和发展建议。     

Response of osteoblasts to low fluid shear stress is time dependent

The process of mechanotransduction of bone, the conversion of a mechanical stimulus into a biochemical response, is known to occur in osteoblasts in response to fluid shear stress. In order to understand the reaction of osteoblasts to various times of flow perfusion, osteoblasts were seeded on three-dimensional scaffolds, and cultured in the following conditions: continuous flow perfusion, intermittent flow perfusion, and static condition. We collected samples on day 4, 8 and 12 for analysis. Osteoblast proliferation was demonstrated by cell proliferation and scanning electron microscopy assay. Additionally, the expression of known markers of differentiation, including alkaline phosphatase and osteocalcin, were tested by qRT-PCR and alkaline phosphatase activity assay, and the deposition of calcium was used as an indicator of mineralization demonstrated by calcium content assay. The results supported that low fluid shear stress plays an important role in the activation of osteoblasts: enhance cell proliferation, increase calcium deposition, and promote the expression of osteoblastic markers. Furthermore, the continuous flow perfusion is a more favorable environment for the initiation of osteoblast activity compared with intermittent flow perfusion. Therefore, the force and time of fluid shear stress are important parameters for osteoblast activation.