Elucidating structure-function relationships from molecule-to-cell-to-tissue: from research modalities to clinical realities

The National Academy of Engineering selected ‘Imaging’ as one of the greatest engineering achievements of the 20th century (Greatest Engineering Achievements of the 20th Century. 2009 (cited 2008, November 10); available from: http://www.greatachievements.org/). The combination of different imaging modalities and technologies for mapping bimolecular and/or biological processes within single cells or even whole organs has extraordinary potential for revolutionizing the diagnosis and treatment of pathophysiological disorders, and thus for mitigating the significant social and economic costs associated with the clinical management of disease. Such integrated imaging approaches will eventually lead to individualized programs for disease prevention through advanced diagnosis, risk stratification and targeted cell therapies resulting in more successful and efficient health care. The goal of this article is to provide readers with a current update of selected of state-of-the-art imaging modalities which would likely to lead to improved clinical outcomes if employed in an integrated approach, including use of ultramicrosensors to detect reactive oxygen/nitrogen species in a single cell, use of electron tomography to visualize and characterize cellular organization in three dimensions (3D), and molecular imaging strategies to assess naturally occurring and therapeutic peripheral and myocardial angiogenesis using targeted radiolabeled tracers

Subnuclear localization, rates and effectiveness of UVC-induced unscheduled DNA synthesis visualized by fluorescence widefield, confocal and super-resolution microscopy

Unscheduled DNA synthesis (UDS) is the final stage of the process of repair of DNA lesions induced by UVC. We detected UDS using a DNA precursor, 5-ethynyl-2′-deoxyuridine (EdU). Using wide-field, confocal and super-resolution fluorescence microscopy and normal human fibroblasts, derived from healthy subjects, we demonstrate that the sub-nuclear pattern of UDS detected via incorporation of EdU is different from that when BrdU is used as DNA precursor. EdU incorporation occurs evenly throughout chromatin, as opposed to just a few small and large repair foci detected by BrdU. We attribute this difference to the fact that BrdU antibody is of much larger size than EdU, and its accessibility to the incorporated precursor requires the presence of denatured sections of DNA. It appears that under the standard conditions of immunocytochemical detection of BrdU only fragments of DNA of various length are being denatured. We argue that, compared with BrdU, the UDS pattern visualized by EdU constitutes a more faithful representation of sub-nuclear distribution of the final stage of nucleotide excision repair induced by UVC. Using the optimized integrated EdU detection procedure we also measured the relative amount of the DNA precursor incorporated by cells during UDS following exposure to various doses of UVC. Also described is the high degree of heterogeneity in terms of the UVC-induced EdU incorporation per cell, presumably reflecting various DNA repair efficiencies or differences in the level of endogenous dT competing with EdU within a population of normal human fibroblasts

Investigating the Optimal Size of Anticancer Nanomedicine

Nanomedicines (NMs) offer new solutions for cancer diagnosis and therapy. However, extension of progression-free interval and overall survival time achieved by Food and Drug Administration-approved NMs remain modest. To develop next generation NMs to achieve superior anticancer activities, it is crucial to investigate and understand the correlation between the physicochemical properties of NMs (particle size in particular) and their interactions with biological systems to establish criteria for NM optimization. Here, we systematically evaluated the size-dependent biological profiles of three monodisperse drug–silica nanoconjugates (NCs; 20, 50, and 200 nm) through both experiments and mathematical modeling and aimed to identify the optimal size for the most effective anticancer drug delivery. Among the three NCs investigated, the 50-nm NC shows the highest tumor tissue retention integrated over time, which is the collective outcome of deep tumor tissue penetration and efficient cancer cell internalization as well as slow tumor clearance, and thus, the highest efficacy against both primary and metastatic tumors in vivo

Heterochromatin protein 1 is recruited to various types of DNA damage

Heterochromatin protein 1 (HP1) family members are chromatin-associated proteins involved in transcription, replication, and chromatin organization. We show that HP1 isoforms HP1-α, HP1-β, and HP1-γ are recruited to ultraviolet (UV)-induced DNA damage and double-strand breaks (DSBs) in human cells. This response to DNA damage requires the chromo shadow domain of HP1 and is independent of H3K9 trimethylation and proteins that detect UV damage and DSBs. Loss of HP1 results in high sensitivity to UV light and ionizing radiation in the nematode Caenorhabditis elegans, indicating that HP1 proteins are essential components of DNA damage response (DDR) systems. Analysis of single and double HP1 mutants in nematodes suggests that HP1 homologues have both unique and overlapping functions in the DDR. Our results show that HP1 proteins are important for DNA repair and may function to reorganize chromatin in response to damage

Distortion in Electrodynamic Loudspeakers Caused by Force Factor Variations

The non linearities in the motor of an electrodynamic loudspeaker are still a discussed topic. This paper studies the influence of the force factor variation with the coil displacement on the harmonic and inter-modulation distortions. The real variation is described at least by a linear and a quadratic term. The effect of each term is studied separately, as they don’t influence the same kind of frequencies, harmonics or inter-modulation. Both terms considered together result in enhanced effects. The dissymmetry of the Bl variation with regard to the coil centered position has also peculiar effects. This paper presents the method developed to calculate the power of each harmonic and inter-modulation frequency. This allows to compare the obtained values and thus the induced nonlinearities