One-step isolation and biochemical characterization of a highlyactive plant PSII monomeric core

We describe a one-step detergent solubilization protocol for isolating a highly active form of Photosystem II (PSII) from Pisum sativum L. Detailed characterization of the preparation showed that the complex was a monomer having no light harvesting proteins attached. This core reaction centre complex had, however, a range of low molecular mass intrinsic proteins as well as the chlorophyll binding proteins CP43 and CP47 and the reaction centre proteins D1 and D2. Of particular note was the presence of a stoichiometric level of PsbW, a low molecular weight protein not present in PSII of cyanobacteria. Despite the high oxygen evolution rate, the core complex did not retain the PsbQ extrinsic protein although there was close to a full complement of PsbO and PsbR and partial level of PsbP. However, reconstitution of PsbP and PsbPQ was possible. The presence of PsbP in absence of LHCII and other chlorophyll a/b binding proteins confirms that LHCII proteins are not a strict requirement for the assembly of this extrinsic polypeptide to the PSII core in contrast with the conclusion of Caffarri et al. (2009)

A proposal for a coordinated effort for the determination of brainwide neuroanatomical connectivity in model organisms at a mesoscopic scale

In this era of complete genomes, our knowledge of neuroanatomical circuitry remains surprisingly sparse. Such knowledge is however critical both for basic and clinical research into brain function. Here we advocate for a concerted effort to fill this gap, through systematic, experimental mapping of neural circuits at a mesoscopic scale of resolution suitable for comprehensive, brain-wide coverage, using injections of tracers or viral vectors. We detail the scientific and medical rationale and briefly review existing knowledge and experimental techniques. We define a set of desiderata, including brain-wide coverage; validated and extensible experimental techniques suitable for standardization and automation; centralized, open access data repository; compatibility with existing resources, and tractability with current informatics technology. We discuss a hypothetical but tractable plan for mouse, additional efforts for the macaque, and technique development for human. We estimate that the mouse connectivity project could be completed within five years with a comparatively modest budget.Comment: 41 page

Recording the Heart Beat of Cattle using a Gradiometer System of Optically Pumped Magnetometers

Monitoring of heart rate has the potential to provide excellent data for the remote monitoring of animals, and heart rate has been associated with stress, pyrexia, pain and illness in animals. However monitoring of heart rate in domesticated animals is difficult as it entails the restraint of the animal (which may in turn affect heart rate), and the application of complex monitoring equipment that is either invasive or not practical to implement under commercial farm conditions. Therefore accurate non-invasive automated remote monitoring of heart rate has not been possible in domesticated animals. Biomagnetism associated with muscle and nerve action provides a promising emerging field in medical sensing, but it is currently confined to magnetically-shielded clinical environments. In this study, we use biomagnetic sensing on commercial dairy cattle under farm conditions as a model system to show proof-of-principle for non-contact magnetocardiography (MCG) outside a controlled laboratory environment. By arranging magnetometers in a differential set-up and using purpose-built low-noise electronics, we are able to suppress common mode noise and successfully record the heart rate, the heart beat intervals and the heart beat amplitude. Comparing the MCG signal with simultaneous data recorded using a conventional electrocardiogram (ECG) allowed alignment of the two signals, and was able to match features of the ECG including the P-wave, the QRS complex and the T-wave. This study has shown the potential for MCG to be developed as a non-contact method for the assessment of heart rate and other cardiac attributes in adult dairy cattle. Whilst this study using an animal model showed the capabilities of un-shielded MCG, these techniques also suggest potentially exciting opportunities in human cardiac medicine outside hospital environments

Automatic Robust Neurite Detection and Morphological Analysis of Neuronal Cell Cultures in High-content Screening

Cell-based high content screening (HCS) is becoming an important and increasingly favored approach in therapeutic drug discovery and functional genomics. In HCS, changes in cellular morphology and biomarker distributions provide an information-rich profile of cellular responses to experimental treatments such as small molecules or gene knockdown probes. One obstacle that currently exists with such cell-based assays is the availability of image processing algorithms that are capable of reliably and automatically analyzing large HCS image sets. HCS images of primary neuronal cell cultures are particularly challenging to analyze due to complex cellular morphology. Here we present a robust method for quantifying and statistically analyzing the morphology of neuronal cells in HCS images. The major advantages of our method over existing software lie in its capability to correct non-uniform illumination using the contrast-limited adaptive histogram equalization method; segment neuromeres using Gabor-wavelet texture analysis; and detect faint neurites by a novel phase-based neurite extraction algorithm that is invariant to changes in illumination and contrast and can accurately localize neurites. Our method was successfully applied to analyze a large HCS image set generated in a morphology screen for polyglutaminemediated neuronal toxicity using primary neuronal cell cultures derived from embryos of a Drosophila Huntington’s Disease (HD) model.National Institutes of Health (U.S.) (Grant

Abstracts from the 8th International Conference on cGMP Generators, Effectors and Therapeutic Implications

This work was supported by a restricted research grant of Bayer AG