598 research outputs found

    The Band Excitation Method in Scanning Probe Microscopy for Rapid Mapping of Energy Dissipation on the Nanoscale

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    Mapping energy transformation pathways and dissipation on the nanoscale and understanding the role of local structure on dissipative behavior is a challenge for imaging in areas ranging from electronics and information technologies to efficient energy production. Here we develop a novel Scanning Probe Microscopy (SPM) technique in which the cantilever is excited and the response is recorded over a band of frequencies simultaneously rather than at a single frequency as in conventional SPMs. This band excitation (BE) SPM allows very rapid acquisition of the full frequency response at each point (i.e. transfer function) in an image and in particular enables the direct measurement of energy dissipation through the determination of the Q-factor of the cantilever-sample system. The BE method is demonstrated for force-distance and voltage spectroscopies and for magnetic dissipation imaging with sensitivity close to the thermomechanical limit. The applicability of BE for various SPMs is analyzed, and the method is expected to be universally applicable to all ambient and liquid SPMs.Comment: 32 pages, 9 figures, accepted for publication in Nanotechnolog

    Resting respiratory tract dendritic cells preferentially stimulate T helper cell Type 2(Th2) responses and require obligatory cytokine signals for induction of Th1 immunity

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    Consistent with their role in host defense, mature dendritic cells (DCs) from central lymphoid organs preferentially prime for T helper cell type 1 (Th1)-polarized immunity. However, the “default” T helper response at mucosal surfaces demonstrates Th2 polarity, which is reflected in the cytokine profiles of activated T cells from mucosal lymph nodes. This study on rat respiratory tract DCs (RTDCs) provides an explanation for this paradox. We demonstrate that freshly isolated RTDCs are functionally immature as defined in vitro, being surface major histocompatibility complex (MHC) II lo, endocytosishi, and mixed lymphocyte reactionlo, and these cells produce mRNA encoding interleukin (IL)-10. After ovalbumin (OVA)-pulsing and adoptive transfer, freshly isolated RTDCs preferentially stimulated Th2-dependent OVA-specific immunoglobulin (Ig)G1 responses, and antigen-stimulated splenocytes from recipient animals produced IL-4 in vitro. However, preculture with granulocyte/macrophage colony stimulating factor increased their in vivo IgG priming capacity by 2–3 logs, inducing production of both Th1- and Th2-dependent IgG subclasses and high levels of IFN-γ by antigen-stimulated splenocytes. Associated phenotypic changes included upregulation of surface MHC II and B7 expression and IL-12 p35 mRNA, and downregulation of endocytosis, MHC II processing– associated genes, and IL-10 mRNA expression. Full expression of IL-12 p40 required additional signals, such as tumor necrosis factor α or CD40 ligand. These results suggest that the observed Th2 polarity of the resting mucosal immune system may be an inherent property of the resident DC population, and furthermore that mobilization of Th1 immunity relies absolutely on the provision of appropriate microenvironmental costimuli

    Magnetic force microscopy of single crystal magnetite (Fe3O4)(abstract)

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    The micromagnetic domain structure of a magnetite (Fe3O4) single crystal has been studied using a magnetic force microscope (MFM). The MFM responds to the perpendicular component of the stray field above the magnetite surface. The sample was polished in the (011) plane. In this case, there are two easy magnetic axes parallel to the surface. Surface domains observed near cracks and edges have a complex closure structure (see Fig. 1), while walls seen far from such boundaries have a sinusoidal structure. Of particular interest is the presence of walls with either even or odd symmetry of the perpendicular stray field component across the transition. These can be conventionally modeled as Bloch or Neel walls, respectively. Both types of walls have been modeled and compared with the experimentally observed structures. We find the Bloch domain walls to be about 300 nm wide, nearly twice the value expected from bulk wall calculations. This distinction is consistent with a surface broadening of the domain wall due to magnetostatic effects.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/70207/2/JAPIAU-75-10-6892-1.pd

    Dual-Frequency Resonance-Tracking Atomic Force Microscopy

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    A dual-excitation method for resonant-frequency tracking in scanning probe microscopy based on amplitude detection is developed. This method allows the cantilever to be operated at or near resonance for techniques where standard phase locked loops are not possible. This includes techniques with non-acoustic driving where the phase of the driving force is frequency and/or position dependent. An example of the later is Piezoresponse Force Microscopy (PFM), where the resonant frequency of the cantilever is strongly dependent on the contact stiffness of the tip-surface junction and the local mechanical properties, but the spatial variability of the drive phase rules out the use of a phase locked loop. Combined with high-voltage switching and imaging, dual-frequency, resonance-tracking PFM allows reliable studies of electromechanical and elastic properties and polarization dynamics in a broad range of inorganic and biological systems, and is illustrated using lead zirconate-titanate, rat tail collagen, and native and switched ferroelectric domains in lithium niobate

    Sagitol C, a new cytotoxic pyridoacridine alkaloid from the sponge Oceanapia sp.

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    AbstractA new pyridoacridine alkaloid named sagitol C (2) together with two known compounds; kuanoniamine C (1) and sagitol (3) were isolated from the EtOAc fraction of the Indonesian sponge Oceanapia sp. Their chemical structures were established on the basis of physical and spectroscopic methods 1D and 2D NMR, in addition to mass spectrometry and comparison with literature data. Sagitol C was found to exhibit cytotoxic activity when tested against different cancer cell lines

    Impact of glycan nature on structure and viscoelastic properties of glycopeptide hydrogels

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    Mucus is a complex biological hydrogel that acts as a barrier for almost everything entering or exiting the body. It is therefore of emerging interest for biomedical and pharmaceutical applications. Besides water, the most abundant components are the large and densely glycosylated mucins, glycoproteins of up to 20 MDa and carbohydrate content of up to 80 wt%. Here, we designed and explored a library of glycosylated peptides to deconstruct the complexity of mucus. Using the well-characterized hFF03 coiled-coil system as a hydrogel-forming peptide scaffold, we systematically probed the contribution of single glycans to the secondary structure as well as the formation and viscoelastic properties of the resulting hydrogels. We show that glycan-decoration does not affect α-helix and coiled-coil formation while it alters gel stiffness. By using oscillatory macrorheology, dynamic light scattering microrheology, and fluorescence lifetime-based nanorheology, we characterized the glycopeptide materials over several length scales. Molecular simulations revealed that the glycosylated linker may extend into the solvent, but more frequently interacts with the peptide, thereby likely modifying the stability of the self-assembled fibers. This systematic study highlights the interplay between glycan structure and hydrogel properties and may guide the development of synthetic mucus mimetics

    The mycotoxin phomoxanthone A disturbs the form and function of the inner mitochondrial membrane.

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    Mitochondria are cellular organelles with crucial functions in the generation and distribution of ATP, the buffering of cytosolic Ca2+ and the initiation of apoptosis. Compounds that interfere with these functions are termed mitochondrial toxins, many of which are derived from microbes, such as antimycin A, oligomycin A, and ionomycin. Here, we identify the mycotoxin phomoxanthone A (PXA), derived from the endophytic fungus Phomopsis longicolla, as a mitochondrial toxin. We show that PXA elicits a strong release of Ca2+ from the mitochondria but not from the ER. In addition, PXA depolarises the mitochondria similarly to protonophoric uncouplers such as CCCP, yet unlike these, it does not increase but rather inhibits cellular respiration and electron transport chain activity. The respiration-dependent mitochondrial network structure rapidly collapses into fragments upon PXA treatment. Surprisingly, this fragmentation is independent from the canonical mitochondrial fission and fusion mediators DRP1 and OPA1, and exclusively affects the inner mitochondrial membrane, leading to cristae disruption, release of pro-apoptotic proteins, and apoptosis. Taken together, our results suggest that PXA is a mitochondrial toxin with a novel mode of action that might prove a useful tool for the study of mitochondrial ion homoeostasis and membrane dynamics
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