An ATM and ATR dependent checkpoint inactivates spindle assembly by targeting CEP63

The effects of ATM and ATR signalling induced by chromosomal breakage have been described extensively in modulating cell cycle progression up to the onset of mitosis. However, DNA damage checkpoint responses in mitotic cells are not well understood. This thesis reports on the effects of double strand breaks on the progression of mitosis. We found ATM and ATR activation can occur in mitotic Xenopus laevis egg extract and the induction of ATM and ATR by chromosomal breakages inhibits spindle assembly in both Xenopus egg extract and somatic cells. The delay in mitotic progression induced by ATM and ATR was found not to involve major spindle assembly factors activities such as, Cdk1, Plx1 and RCC1/Ran-GTP. However, normal anastral spindles formation around linear DNA coated beads, which can activate ATM and ATR, linked centrosome-driven spindle assembly to ATM and ATR dependent spindle defects. cDNA expression library screening was undertaken to identify novel ATM and ATR targets in this mitotic checkpoint pathway, through which the novel centrosomal protein XCEP63 was identified as a likely candidate. Data obtained from depletion and reconstitution of XCEP63 in Xenopus egg extract established that normal centrosome-driven spindle assembly requires XCEP63. Moreover, ATM and ATR phosphorylates XCEP63 on serine 560 and promotes delocalisation from the centrosome. ATM and ATR inhibition or addition of non-phosphorylable XCEP63 recombinant protein mutated at serine 560 prevents spindle assembly abnormalities. These findings suggest that ATM and ATR regulate mitotic events by targeting XCEP63 and centrosome-dependent spindle assembly. This pathway may provide support for DNA repair processes or regulate cell survival in the presence of mitotic DNA damage

Kinetic and thermodynamic analysis of leech-derived tryptase inhibitor interaction with bovine tryptase and bovine trypsin

The interaction of leech-derived tryptase inhibitor (LDTI) with bovine liver capsule tryptase (BLCT) and bovine trypsin has been studied using both thermodynamic and kinetic approaches. Several differences were detected: (i) the equilibrium affinity of LDTI for BLCT (K-a = 8.9 x 10(5) M-1) is about 600-fold lower than that for bovine trypsin (K-a = 5.1 x 10(8) M-1); (ii) LDTI behaves as a purely non-competitive inhibitor of BLCT, while it is a purely competitive inhibitor of bovine trypsin. These functional data are compared with those previously reported for the LDTI binding to human tryptase, where tight inhibition occurs at two of the four active sites of the tetramer (K-a = 7.1 x 10(8) M-1). Amino acid sequence alignment of BLCT, human beta II-tryptase and bovine trypsin allows us to infer some possible structural basis for the observed functional differences

Recording, monitoring and managing the conservation of historic sites : a new application for BGS Sigma

Historic Environment Scotland (HES), a non-departmental public body of the Scottish Government charged with safeguarding the nation’s historic environment, is directly responsible for 335 sites of national significance, most of which are built from stone. Similar to other heritage organisations, HES needs a system that can store and present conservation and maintenance information for historic sites; ideally, the same system could be used to plan effective programmes of maintenance and repair. To meet this need, the British Geological Survey (BGS) has worked with HES to develop an integrated digital site assessment system that provides a refined survey process for stone-built (and other) historic sites. Based on the BGS System for Integrated Geoscience Mapping (BGS▪SIGMA)—an integrated workflow underpinned by a geo-spatial platform for data capture and interpretation—the system is built on top of ESRI’s ArcGIS software, and underpinned by a relational database. Users can populate custom-built data entry forms to record maintenance issues and repair specifications for architectural elements ranging from individual blocks of stone to entire building elevations. Photographs, sketches, and digital documents can be linked to architectural elements to enhance the usability of the data. Predetermined data fields and supporting dictionaries constrain the input parameters to ensure a high degree of consistency and facilitate data extraction and querying. Presenting the data within a GIS provides a versatile planning tool for scheduling works, specifying materials, identifying skills needed for repairs, and allocating resources. The overall condition of a site can be monitored accurately over time by repeating the survey at regular intervals (e.g. every 5 years). Other datasets can be linked to the database and other geospatially referenced datasets can be superimposed in GIS, adding considerably to the scope and utility of the system. The system can be applied to any geospatially referenced object in a wide range of situations thus providing many potential applications in conservation, archaeology and related fields

How to design and evaluate interventions to improve outcomes for patients with multimorbidity

Multimorbidity is a major challenge for patients and healthcare providers. The limited evidence of the effectiveness of interventions for people with multimorbidity means that there is a need for much more research and trials of potential interventions. Here we present a consensus view from a group of international researchers working to improve care for people with multimorbidity to guide future studies of interventions. We suggest that there is a need for careful consideration of whom to include, how to target interventions that address specific problems and that do not add to treatment burden, and selecting outcomes that matter both to patients and the healthcare system. Innovative design of these interventions will be necessary as many will be introduced in service settings and it will be important to ensure methodological rigour, relevance to service delivery, and generalizability across healthcare systems

Long-term growth in vitro of isolated, fully differentiated neurones from the central nervous system of an adult insect

A method is described for the isolation and growth in vitro of fully differentiated neurones from the thoracic ganglia of adult cockroaches. The presence of insect blood in the culture system is shown to promote growth. The morphology of the growing neurones and the plasticity of the branching processes are described and growth rates are measured. Using a fluorescent Ca2+ indicator dye, changes of intracellular calcium levels in the growing neurones in response to K+ depolarization have been measured. The results, indicating the presence of voltage-dependent Ca2+ channels on neuronal processes in vitro, show that neurones can be maintained in a functional state for several weeks by this technique. Such preparations could prove useful for studying a variety of physiological and pharmacological properties of neurones, including the mechanisms controlling growth, synapse formation and neuronal interactions with other cell types. <br/

N, P and K budgets for crop rotations on nine organic farms in the UK

On organic farms, where the importation of materials to build/maintain soil fertility is restricted, it is important that a balance between inputs and outputs of nutrients is achieved to ensure both short-term productivity and long-term sustainability. This paper considers different approaches to nutrient budgeting on organic farms and evaluates the sources of bias in the measurements and/or estimates of the nutrient inputs and outputs. The paper collates 88 nutrient budgets compiled at the farm scale in 9 temperate countries. All the nitrogen (N) budgets showed an N surplus (average 83.2 kg N ha-1 year-1). The efficiency of N use, defined as outputs/inputs, was highest (0.9) and lowest (0.2) in arable and beef systems respectively. The phosphorus (P) and potassium (K) budgets showed both surpluses and deficits (average 3.6 kg P ha-1 year-1, 14.2 kg K ha-1 year-1) with horticultural systems showing large surpluses resulting from purchased manure. The estimation of N fixation and quantities of nutrients in purchased manures may introduce significant errors in nutrient budgets. Overall, the data illustrate the diversity of management systems in place on organic farms, and suggest that used together with soil analysis, nutrient budgets are a useful tool for improving the long-term sustainability of organic systems

Abundance and movements of caribou in the oilfield complex near Prudhoe Bay, Alaska

We examined the distribution and movements of 141 radiocollared female caribou (Rangifer tarandus granti) of the Central Arctic Herd during summer, 1980-1993. Numbers of caribou locations within each of 5 quadrats along the arctic coast were totalled separately for days during which insects were active and inactive, and numbers of east-west and west-east crossings of each quadrat mid-line were determined from sequential observations. Both abundance and lateral movements of radiocollared females in the quadrat encompassing the intensively-developed Prudhoe Bay oilfield complex were significantly lower than in other quadrats (P &lt; 0.001 and P &lt; 0.00001, respectively). Avoidance of, and fewer movements within, the complex by female caribou are ostensibly in response to the dense network of production and support facilities, roads, above-ground pipelines, and the associated vehicular and human activity. Impaired access to this area constitutes a functional loss of habitat

Modelling mucociliary clearance

Mathematical modelling of the fluid mechanics of mucociliary clearance (MCC) is reviewed and future challenges for researchers are discussed. The morphology of the bronchial and tracheal airway surface liquid (ASL) and ciliated epithelium are briefly introduced. The cilia beat cycle, beat frequency and metachronal coordination are described, along with the rheology of the mucous layer. Theoretical modelling of MCC from the late 1960s onwards is reviewed, and distinctions between ‘phenomenological’, ‘slender body theory’ and recent ‘fluid–structure interaction’ models are explained.\ud \ud The ASL consists of two layers, an overlying mucous layer and underlying watery periciliary layer (PCL) which bathes the cilia. Previous models have predicted very little transport of fluid in the PCL compared with the mucous layer. Fluorescent tracer transport experiments on human airway cultures conducted by Matsui et al. [Matsui, H., Randell, S.H., Peretti, S.W., Davis, C.W., Boucher, R.C., 1998. Coordinated clearance of periciliary liquid and mucus from airway surfaces. J. Clin. Invest. 102 (6), 1125–1131] apparently showed equal transport in both the PCL and mucous layer. Recent attempts to resolve this discrepancy by the present authors are reviewed, along with associated modelling findings. These findings have suggested new insights into the interaction of cilia with mucus due to pressure gradients associated with the flat PCL/mucus interface. This phenomenon complements previously known mechanisms for ciliary propulsion. Modelling results are related to clinical findings, in particular the increased MCC observed in patients with pseudohypoaldosteronism. Recent important advances by several groups in modelling the fluid–structure interaction by which the cilia movement and fluid transport emerge from specification of internal mechanics, viscous and elastic forces are reviewed. Finally, we discuss the limitations of existing work, and the challenges for the next generation of models, which may provide further insight into this complex and vital system