Preferential localization of a vesicular monoamine transporter to dense core vesicles in PC12 cells.

Neurons and endocrine cells have two types of secretory vesicle that undergo regulated exocytosis. Large dense core vesicles (LDCVs) store neural peptides whereas small clear synaptic vesicles store classical neurotransmitters such as acetylcholine, gamma-aminobutyric acid (GABA), glycine, and glutamate. However, monoamines differ from other classical transmitters and have been reported to appear in both LDCVs and smaller vesicles. To localize the transporter that packages monoamines into secretory vesicles, we have raised antibodies to a COOH-terminal sequence from the vesicular amine transporter expressed in the adrenal gland (VMAT1). Like synaptic vesicle proteins, the transporter occurs in endosomes of transfected CHO cells, accounting for the observed vesicular transport activity. In rat pheochromocytoma PC12 cells, the transporter occurs principally in LDCVs by both immunofluorescence and density gradient centrifugation. Synaptic-like microvesicles in PC12 cells contain relatively little VMAT1. The results appear to account for the storage of monoamines by LDCVs in the adrenal medulla and indicate that VMAT1 provides a novel membrane protein marker unique to LDCVs

Biomass burning emisison inventory with daily resolution: Application to aircraft observations of Asian outflow

We develop a daily-resolved global emission inventory for biomass burning using AVHRR satellite observations of fire activity corrected for data gaps and scan angle biases. We implemented this inventory in a global three-dimensional model (GEOSCHEM) to simulate aircraft CO observations during the TRACE-P mission over the NW Pacific in February-April 2001. Seasonal biomass burning in SE Asia was a major contributor to the outflow of Asian pollution observed in TRACE-P and shows large day-to-day fluctuations that vary depending on location. Three simulations were conducted with the same 3-month total (February-April) emissions but different temporal distributions: 2001 daily resolved, 2001 monthly resolved, and climatological monthly resolved. The effect of daily resolved versus monthly resolved 2001 emissions in the simulation of CO is less than 8 ppbv in Asian outflow over the NW Pacific but can exceed 100 ppbv over source regions. The relatively small effect in Asian outflow reflects spatial and temporal averaging of emissions during ageing in the continental boundary layer. Significant improvement in the simulation of TRACE-P observations (as diagnosed by the resolved variance) is found when using 2001 monthly versus climatological monthly emissions, but using 2001 daily emissions does not offer further improvement. Copyright 2003 by the American Geophysical Union

3D quantitative imaging of unprocessed live tissue reveals epithelial defense against bacterial adhesion and subsequent traversal requires MyD88.

While a plethora of in vivo models exist for studying infectious disease and its resolution, few enable factors involved in the maintenance of health to be studied in situ. This is due in part to a paucity of tools for studying subtleties of bacterial-host interactions at a cellular level within live organs or tissues, requiring investigators to rely on overt outcomes (e.g. pathology) in their research. Here, a suite of imaging technologies were combined to enable 3D and temporal subcellular localization and quantification of bacterial distribution within the murine cornea without the need for tissue processing or dissection. These methods were then used to demonstrate the importance of MyD88, a central adaptor protein for Toll-Like Receptor (TLR) mediated signaling, in protecting a multilayered epithelium against both adhesion and traversal by the opportunistic bacterial pathogen Pseudomonas aeruginosa ex vivo and in vivo

Algal Viruses: The (Atomic) Shape of Things to Come

This is the final version. Available on open access from MDPI via the DOI in this recordVisualization of algal viruses has been paramount to their study and understanding. The direct observation of the morphological dynamics of infection is a highly desired capability and the focus of instrument development across a variety of microscopy technologies. However, the high temporal (ms) and spatial resolution (nm) required, combined with the need to operate in physiologically relevant conditions presents a significant challenge. Here we present a short history of virus structure study and its relation to algal viruses and highlight current work, concentrating on electron microscopy and atomic force microscopy, towards the direct observation of individual algae⁻virus interactions. Finally, we make predictions towards future algal virus study direction with particular focus on the exciting opportunities offered by modern high-speed atomic force microscopy methods and instrumentation.The previously unpublished AFM images shown in this manuscript were generated in the Plymouth Marine Laboratory and University of Exeter Environmental Single Cell Genomics Facility which was supported by the UK’s Natural Environment Research Council (NERC) and The Wolfson Foundation. C.T.E. is supported by an EPSRC and Bristol Nano Dynamics Ltd PhD studentship

Improving tuberculosis diagnosis: Better tests or better healthcare?

In a Perspective accompanying Sylvia and colleagues, Carlton Evans and colleagues discuss the challenge of squaring policies around tuberculosis diagnosis with the realities of clinical practice in small villages and low-resource settings

Novel Characteristics of Valveless Pumping

This study investigates the occurrence of valveless pumping in a fluidfilled system consisting of two open tanks connected by an elastic tube. We show that directional flow can be achieved by introducing a periodic pinching applied at an asymmetrical location along the tube, and that the flow direction depends on the pumping frequency. We propose a relation between wave propagation velocity, tube length, and resonance frequencies associated with shifts in the pumping direction using numerical simulations. The eigenfrequencies of the system are estimated from the linearized system, and we show that these eigenfrequencies constitute the resonance frequencies and the horizontal slope frequencies of the system; 'horizontal slope frequency' being a new concept. A simple model is suggested, explaining the effect of the gravity driven part of the oscillation observed in response to the tank and tube diameter changes. Results are partly compared with experimental findings.Art. no. 22450

Visualising muscle anatomy using three-dimensional computer models - an example using the head and neck muscles of Sphenodon

We demonstrate how the computer-based technique of multi-body dynamics analysis (MDA) can be used to create schematic, but informative three-dimensional (3D) representations of complex muscle anatomy. As an example we provide an overview of the head and neck muscles present in Sphenodon (Diapsida: Lepidosauria: Rhynchocephalia). First a computer model based on micro-computed tomography datasets provides a detailed and anatomically correct three-dimensional (3D) framework to work from. Secondly, muscles are represented by groups of cylinders that can be colour coded as desired. This allows muscle positions, attachment areas, and 3D orientation to be visualised clearly. This method has advantages over imaging techniques such as two-dimensional drawings and permits the form and function of the muscles to be understood in a way that is not always possible with more classical visualisation techniques. Copyright: Palaeontological Association December 2009