Cre-Dependent Expression of Multiple Transgenes in Isolated Neurons of the Adult Forebrain

Background: Transgenic mice with mosaic, Golgi-staining-like expression of enhanced green fluorescent protein (EGFP) have been very useful in studying the dynamics of neuronal structure and function. In order to further investigate the molecular events regulating structural plasticity, it would be useful to express multiple proteins in the same sparse neurons, allowing co-expression of functional proteins or co-labeling of subcellular compartments with other fluorescent proteins. However, it has been difficult to obtain reproducible expression in the same subset of neurons for direct comparison of neurons expressing different functional proteins. Principal Findings: Here we describe a Cre-transgenic line that allows reproducible expression of transgenic proteins of choice in a small number of neurons of the adult cortex, hippocampus, striatum, olfactory bulb, subiculum, hypothalamus, superior colliculus and amygdala. We show that using these Cre-transgenic mice, multiple Cre-dependent transgenes can be expressed together in the same isolated neurons. We also describe a Cre-dependent transgenic line expressing a membrane associated EGFP (EGFP-F). Crossed with the Cre-transgenic line, EGFP-F expression starts in the adolescent forebrain, is present in dendrites, dendritic protrusions, axons and boutons and is strong enough for acute or chronic in vivo imaging. Significance: This triple transgenic approach will aid the morphological and functional characterization of neurons in various Cre-dependent transgenic mice

A planet within the debris disk around the pre-main-sequence star AU Microscopii

AU Microscopii (AU Mic) is the second closest pre main sequence star, at a distance of 9.79 parsecs and with an age of 22 million years. AU Mic possesses a relatively rare and spatially resolved3 edge-on debris disk extending from about 35 to 210 astronomical units from the star, and with clumps exhibiting non-Keplerian motion. Detection of newly formed planets around such a star is challenged by the presence of spots, plage, flares and other manifestations of magnetic activity on the star. Here we report observations of a planet transiting AU Mic. The transiting planet, AU Mic b, has an orbital period of 8.46 days, an orbital distance of 0.07 astronomical units, a radius of 0.4 Jupiter radii, and a mass of less than 0.18 Jupiter masses at 3 sigma confidence. Our observations of a planet co-existing with a debris disk offer the opportunity to test the predictions of current models of planet formation and evolution.Comment: Nature, published June 24th [author spelling name fix

Reconstruction and simulation of neocortical microcircuitry

We present a first-draft digital reconstruction of the microcircuitry of somatosensory cortex of juvenile rat. The reconstruction uses cellular and synaptic organizing principles to algorithmically reconstruct detailed anatomy and physiology from sparse experimental data. An objective anatomical method defines a neocortical volume of 0.29 ± 0.01 mm3 containing ∼31,000 neurons, and patch-clamp studies identify 55 layer-specific morphological and 207 morpho-electrical neuron subtypes. When digitally reconstructed neurons are positioned in the volume and synapse formation is restricted to biological bouton densities and numbers of synapses per connection, their overlapping arbors form ∼8 million connections with ∼37 million synapses. Simulations reproduce an array of in vitro and in vivo experiments without parameter tuning. Additionally, we find a spectrum of network states with a sharp transition from synchronous to asynchronous activity, modulated by physiological mechanisms. The spectrum of network states, dynamically reconfigured around this transition, supports diverse information processing strategies

Additive manufacturing methods for pharmaceutical and medical applications

Three-dimensional (3D) printing has been geared up to become a highly revolutionary technology within the manufacturing industry, especially within the pharmaceutical and medical fields. Three-dimensional printing utilizes computer-aided design (CAD) software to manufacture objects of any size and shape. Due to the potential capabilities, many industries are now making use of this manufacturing technique, from rapid prototyping in engineering to the production of personalized medicine within the medical and pharmaceutical industries. It has the potential to cause a paradigm shift and revolutionize the way medicines and medical devices are designed, manufactured, and used. This chapter provides an overview of the field of 3D printing, current medical applications, 3D-printed dosage forms, and drug delivery systems as well as the benefits and limitations of this technique. It focuses on the potential and motivations of this method of additive manufacturing, including the challenges and future possibilities it could bring to the field of medicine and pharmaceuticals.<br/

Poly(caprolactone)/lignin-based 3D-printed dressings loaded with a novel combination of bioactive agents for wound-healing applications

Curcumin (CUR) has been shown to possess significant anti-inflammatory properties and significant wound healing potential. Additionally, lignin (LIG) is a renewable biomacromolecule with well-known antioxidant and antimicrobial properties, which makes this biomacromolecule a good candidate to be included in medical materials, such as wound dressings. Although many of the wound dressings used at present have interesting features, some are limited in terms of antibacterial properties. To address these limitations, in the present work, both CUR and LIG were combined with poly(caprolactone) (PCL), a biocompatible polymer, to obtain dressings with antioxidant and antimicrobial properties for wound healing treatment. Moreover, D-Panthenol (DPA) was included in the composite materials formulation due to its skin regenerative ability by enhancing epidermal differentiation. Semi-solid extrusion (SSE) 3D printing was used to manufacture wound dressings without the use of any solvents. 3D-printed dressings provided a sustained DPA and CUR release for periods of up to 4 and 35 days, respectively. A DPPH (2,2-diphenyl-1-picrylhydrozyl) assay was performed confirming that the presence of LIG and CUR provided antioxidant properties to the 3D-printed dressings. Additionally, these 3D-printed materials showed a marked resistance to adherence of Staphylococcus aureus when compared to the PCL control 3D-printed samples, resulting in substantial reductions of up to 89.9% and 98.9% after incubation periods of 4 h and 24 h respectively. Although, all of the 3D-printed materials were able to provide a supportive environment for cellular attachment, viability and growth, the combination of both bioactive compounds CUR and DPA exhibited the most significant values for cell viability and proliferation. In vivo wound healing study performed in Wistar rats showed that dressings containing these novel two compounds CUR and DPA exhibited marked improvement at any stage of the treatment process. Finally, histological examination revealed that dressings loaded with CUR and DPA also showed the best outcomes for all the evaluated parameters: (i) epithelisation, (ii) inflammatory reaction, (iii) proliferation rate of fibroblast and (iv) neoangiogenesis.Wellcome Trust UNS4004

A framework for examining leadership in extreme contexts

In this review, we develop a framework to guide future research and to examine the execution of leadership in extreme contexts. We start by defining and distinguishing extreme contexts from crisis and other contexts. A five component typology is developed comprised of magnitude of consequences, form of threat, probability of consequences, location in time and physical or psychological–social proximity. We discuss the unique influences these components have on leadership processes in extreme contexts examining the relevance of organization types such as critical action and high reliability organizations. Further, we present a set of factors that may attenuate or intensify the dimensions comprising an extreme context, thus influencing either a team or organization\u27s ability for adaptive leadership response. Ultimately this framework seeks to develop a richer understanding of extreme contexts to advance the future development of contextualized theories of leadership for extreme contexts