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Large, ethidium bromide-loaded liposomes electrically pulsed in the presence of externally added DNA display the bright fluorescence of DNA-ethidium bromide complexes. Sonication of these liposomes increases the fluorescence of trapped DNA-ethidium bromide complexes by no more than about 40%. These results are thus in agreement with a mechanism involving electropores for DNA uptake but do not support an alternative mechanism, invoking invagination and pinching-off of the lipid bilayer, through which internalized DNA is shielded from the liposome contents.  相似文献   
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Central to understanding mechanotransduction in the knee meniscus is the characterization of meniscus cell mechanics. In addition to biochemical and geometric differences, the inner and outer regions of the meniscus contain cells that are distinct in morphology and phenotype. This study investigated the regional variation in meniscus cell mechanics in comparison with articular chondrocytes and ligament cells. It was found that the meniscus contains two biomechanically distinct cell populations, with outer meniscus cells being stiffer (1.59 ± 0.19 kPa) than inner meniscus cells (1.07 ± 0.14 kPa). Additionally, it was found that both outer and inner meniscus cell stiffnesses were similar to ligament cells (1.32 ± 0.20 kPa), and articular chondrocytes showed the highest stiffness overall (2.51 ± 0.20 kPa). Comparison of compressibility characteristics of the cells showed similarities between articular chondrocytes and inner meniscus cells, as well as between outer meniscus cells and ligament cells. These results show that cellular biomechanics vary regionally in the knee meniscus and that meniscus cells are biomechanically similar to ligament cells. The mechanical properties of musculoskeletal cells determined in this study may be useful for the development of mathematical models or the design of experiments studying mechanotransduction in a variety of soft tissues.  相似文献   
135.
The dynamics of isogenic cell populations can be described by cell population balance models that account for phenotypic heterogeneity. To utilize the predictive power of these models, however, we must know the rates of single-cell reaction and division and the bivariate partition probability density function. These three intrinsic physiological state (IPS) functions can be obtained by solving an inverse problem that requires knowledge of the phenotypic distributions for the overall cell population, the dividing cell subpopulation and the newborn cell subpopulation. We present here a robust computational procedure that can accurately estimate the IPS functions for heterogeneous cell populations. A detailed parametric analysis shows how the accuracy of the inverse solution is affected by discretization parameters, the type of non-parametric estimators used, the qualitative characteristics of phenotypic distributions and the unknown partitioning probability density function. The effect of finite sampling and measurement errors on the accuracy of the recovered IPS functions is also assessed. Finally, we apply the procedure to estimate the IPS functions of an E. coli population carrying an IPTG-inducible genetic toggle network. This study completes the development of an integrated experimental and computational framework that can become a powerful tool for quantifying single-cell behavior using measurements from heterogeneous cell populations.  相似文献   
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A 78 year-old female patient underwent a total mastectomy with axillary lymph node dissection for a primary breast osteosarcoma. Microscopically the tumor was identical to grade II skeletal osteosarcoma. Immunohistochemically no reactivity was detected, either for the epithelial markers EMA, AE1/AE3, CK8, 18, 19, or for HER-2/neu, estrogen and progesterone receptors, as well as fluorescent IN SITU hybridization for HER-2/neu. The diagnosis of this tumor fulfills certain clinicopathological criteria. Mammary osteosarcoma is usually developed in phyllodes tumors or carcinosarcomas of the breast as a result of metaplasia of the epithelial component. This rare tumor of the breast is occasionally associated with prior radiation therapy or well documented trauma. Mammary osteosarcoma is a biologically aggressive neoplasm with a 38% five-year survival rate. Surgical resection is the most effective therapy to date. Adjuvant treatment -chemotherapy or radiotherapy- has shown no clear benefit. An extensive review of the literature is also presented.  相似文献   
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The way in which the nucleus experiences mechanical forces has important implications for understanding mechanotransduction. Knowledge of nuclear material properties and, specifically, their relationship to the properties of the bulk cell can help determine if the nucleus directly experiences mechanical loads, or if it is a signal transduction mechanism secondary to cell membrane deformation that leads to altered gene expression. Prior work measuring nuclear material properties using micropipette aspiration suggests that the nucleus is substantially stiffer than the bulk cell [Guilak, F., Tedrow, J.R., Burgkart, R., 2000. Viscoelastic properties of the cell nucleus. Biochem. Biophys. Res. Commun. 269, 781–786], whereas recent work with unconfined compression of single chondrocytes showed a nearly one-to-one correlation between cellular and nuclear strains [Leipzig, N.D., Athanasiou, K.A., 2008. Static compression of single chondrocytes catabolically modifies single-cell gene expression. Biophys. J. 94, 2412–2422]. In this study, a linearly elastic finite element model of the cell with a nuclear inclusion was used to simulate the unconfined compression data. Cytoplasmic and nuclear stiffnesses were varied from 1 to 7 kPa for several combinations of cytoplasmic and nuclear Poisson's ratios. It was found that the experimental data were best fit when the ratio of cytoplasmic to nuclear stiffness was 1.4, and both cytoplasm and nucleus were modeled as incompressible. The cytoplasmic to nuclear stiffness ratio is significantly lower than prior reports for isolated nuclei. These results suggest that the nucleus may behave mechanically different in situ than when isolated.  相似文献   
139.
The cytoskeleton is known to play an important role in the biomechanical nature and structure of cells, but its particular function in compressive characteristics has not yet been fully examined. This study focused on the contribution of the main three cytoskeletal elements to the bulk compressive stiffness (as measured by the compressive modulus), volumetric or apparent compressibility changes (as further indicated by apparent Poisson's ratio), and recovery behavior of individual chondrocytes. Before mechanical testing, cytochalasin D, acrylamide, or colchicine was used to disrupt actin microfilaments, intermediate filaments, or microtubules, respectively. Cells were subjected to a range of compressive strains and allowed to recover to equilibrium. Analysis of the video recording for each mechanical event yielded relevant compressive properties and recovery characteristics related to the specific cytoskeletal disrupting agent and as a function of applied axial strain. Inhibition of actin microfilaments had the greatest effect on bulk compressive stiffness (∼50% decrease compared to control). Meanwhile, intermediate filaments and microtubules were each found to play an integral role in either the diminution (compressibility) or retention (incompressibility) of original cell volume during compression. In addition, microtubule disruption had the largest effect on the “critical strain threshold” in cellular mechanical behavior (33% decrease compared to control), as well as the characteristic time for recovery (∼100% increase compared to control). Elucidating the role of the cytoskeleton in the compressive biomechanical behavior of single cells is an important step toward understanding the basis of mechanotransduction and the etiology of cellular disease processes.  相似文献   
140.
Ioannou  Kyriacos  Vlasiou  Manos C. 《Biometals》2022,35(4):639-652
BioMetals - The first appearance of SARS-CoV-2 is dated back to 2019. This new member of the coronavirus family has caused more than 5 million deaths worldwide up until the end of January 2022. At...  相似文献   
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