Predicting the Remaining Lifespan and Cultivation-Related Loss of Osteogenic Capacity of Bone Marrow Multipotential Stromal Cells Applicable across a Broad Donor Age Range

Background and Objectives. Culture expanded multipotential stromal cells (MSCs) have considerable potential for bone regeneration therapy but their wider use is constrained by the lack of simple and predictive assays of functional potency. Extended passaging leads to loss of multipotency but speed of decline depends on MSC donor age. The aim of this study was to develop an assay predictive of MSC culture longevity applicable to a broad donor age range. Materials and Methods. Bone marrow (BM, =7) was obtained from a diverse range (2–72 years) of healthy donors. MSCs were culture expanded to senescence and their osteoprogenitor content, gene expression profiles, epigenetic signature, and telomere behaviour were measured throughout. Output data was combined for modelling purposes. Results. Regardless of donor age, cultures’ osteoprogenitor content correlated better with remaining lifespan (population doublings before senescence, PD-BS) than proliferative history (accrued PDs). Individual gene’s expression or telomere length did not predict PD-BS but methylation of individual CpG islands did, PRAMEF2 in particular ( = 0.775). Coupling the steep relationship of relative SPARC expression with PD-BS ( = −0.753) the formula SPARC × 1/PREMEF2 gave an improved correlation ( = −0.893).Conclusion. A formula based on SPARC mRNA and PRAMEF2 methylation may be used to predict remaining BM-MSC longevity and related loss of multipotentiality independent of donor age

Portable LED fluorescence instrumentation for the rapid assessment of potable water quality

Characterising the organic and microbial matrix of water are key issues in ensuring a safe potable water supply. Current techniques only confirm water quality retrospectively via laboratory analysis of discrete samples. Whilst such analysis is required for regulatory purposes, it would be highly beneficial to monitor water quality in-situ in real time, enabling rapid water quality assessment and facilitating proactive management of water supply systems. A novel LED-based instrument, detecting fluorescence peaks C and T (surrogates for organic and microbial matter, respectively), was constructed and performance assessed. Results from over 200 samples taken from source waters through to customer tap from three UK water companies are presented. Excellent correlation was observed between the new device and a research grade spectrophotometer (r 2 = 0.98 and 0.77 for peak C and peak T respectively), demonstrating the potential of providing a low cost, portable alternative fluorimeter. The peak C/TOC correlation was very good (r 2 = 0.75) at low TOC levels found in drinking water. However, correlations between peak T and regulatory measures of microbial matter (2 day/3 day heterotrophic plate counts (HPC), E. coli, and total coliforms) were poor, due to the specific nature of these regulatory measures and the general measure of peak T. A more promising correlation was obtained between peak T and total bacteria using flow cytometry. Assessment of the fluorescence of four individual bacteria isolated from drinking water was also considered and excellent correlations found with peak T (Sphingobium sp. (r 2 = 0.83); Methylobacterium sp. (r 2 = 1.0); Rhodococcus sp. (r 2 = 0.86); Xenophilus sp. (r 2 = 0.96)). It is notable that each of the bacteria studied exhibited different levels of fluorescence as a function of their number. The scope for LED based instrumentation for insitu, real time assessment of the organic and microbial matrix of potable water is clearly demonstrated

The effect of landscape topography and in-stream habitat on the distribution, growth, and survival of Lahontan cutthroat trout (Oncorhynchus clarki henshawi) in a high desert watershed

Lahontan cutthroat trout (LCT) (Oncorhynchus clarki henshawi) are currently limited in their distribution to a patchwork of small isolated populations, the result of habitat degradation and natural variation in landscape and in-stream conditions. The objectives of this study were to determine how landscape level topographic features influence LCT distribution patterns. The work was carried out in a sub-basin of the Quinn River system, McDermitt Creek, which drains the sagebrush desert of southeastern Oregon and northern Nevada. Headwater tributaries of this creek consist of alternating canyon-confined and valley bounded reaches and LCT within these systems are challenged by low discharge and high temperatures during the summer, and anchor ice during the winter. Contiguous whole stream surveys were used to look at trout distribution during the summer of 2003 and spring and fall of 2004. Our results showed that topography affected LCT distribution in all reaches. LCT numbers were highest in areas with greater numbers of nick-points (the transition zones between less confined and more confined valley segments) and greater valley confinement. Additionally, in the downstream portion of our headwater reaches, more LCT were found in nick-points than expected based on the availability of this habitat type. Our data suggest that hyporheic inputs may be high in such areas, thus providing LCT with shelter from warm water in the summer, anchor ice in the winter and shallow stream depths during all seasons. Spatial occurrence of these areas of refugia can be taken into consideration when planning land-use activities and restoration efforts. After almost 30 years of conservation effort, Lahontan cutthroat trout (LCT) (Oncorhynchus clarki henshawi) populations remain in peril throughout the Great Basin region of southeastern Oregon and northern Nevada. This is not surprising since very little research has been done on the ecology of LCT, and management has been carried out without essential information on this sub-species. The objectives of our study were to determine whether there are inter-stream or inter-reach differences in LCT habitat selection, and if those differences corresponded to differences in fish survival and growth. We relied upon contiguous whole stream surveys to look at fish distribution and in-stream habitat. The use of half duplex PIT tags allowed us to determine the growth, movement patterns, and survival rates of most adult (>100 mm) LCT within the system. We found greater trout growth (median daily change in biomass 0.46% vs. 0.07%), but lower survival (35% vs. 61% between July and November), in relatively warmer and more open reaches than in cooler reaches. Additionally, undercut banks (predation shelter) were more important to trout in cooler stream reaches than in warmer ones, where habitat choice responded first to cold water input (which was influenced by topography). Our results will be useful in identifying and describing areas of high quality LCT habitat in low order streams throughout the Great Basin, thus allowing informed management decisions to facilitate the recovery of the sub-species

Phosphorylation of Phosphoenolpyruvate Carboxylase Is Essential for Maximal and Sustained Dark CO2 Fixation and Core Circadian Clock Operation in the Obligate Crassulacean Acid Metabolism Species Kalanchoe fedtschenkoi

Phosphoenolpyruvate carboxylase (PPC; EC 4.1.1.31) catalyzes primary nocturnal CO2 fixation in Crassulacean acid metabolism (CAM) species. CAM PPC is regulated posttranslationally by a circadian clock-controlled protein kinase called phosphoenolpyruvate carboxylase kinase (PPCK). PPCK phosphorylates PPC during the dark period, reducing its sensitivity to feedback inhibition by malate and thus enhancing nocturnal CO2 fixation to stored malate. Here, we report the generation and characterization of transgenic RNAi lines of the obligate CAM species Kalanchoë fedtschenkoi with reduced levels of KfPPCK1 transcripts. Plants with reduced or no detectable dark phosphorylation of PPC displayed up to a 66% reduction in total dark period CO2 fixation. These perturbations paralleled reduced malate accumulation at dawn and decreased nocturnal starch turnover. Loss of oscillations in the transcript abundance of KfPPCK1 was accompanied by a loss of oscillations in the transcript abundance of many core circadian clock genes, suggesting that perturbing the only known link between CAM and the circadian clock feeds back to perturb the central circadian clock itself. This work shows that clock control of KfPPCK1 prolongs the activity of PPC throughout the dark period in K. fedtschenkoi, optimizing CAM-associated dark CO2 fixation, malate accumulation, CAM productivity, and core circadian clock robustness

Occurrence of transients in water distribution networks

The common existence of pressure transients in operational water distribution systems (WDS) requires their characterisation and assessment of their impact. This paper performs such characterisation by evidencing the occurrence and the differences in pressure transient behaviour in complex WDS. Ten samples of continuously recorded high resolution pressures from diverse networks and sources were analysed. The presented pressure traces show regular and occasional pressure transient waves in various complex networks. Histogram analysis of the rate of change of head provides some insight into transient behaviour in these sites. Although there was no distinct correlation between network characteristics (ie. length, diameter, age) and transient behaviour, network complexity was observed to change the transient characteristics. Transient characteristics were observed to be strongly influenced by likely sources, in particular commercial customers. The data highlights the need to understand, quantify and characterise transients and hence link to possible impacts, such as structural or water quality failures

Assessment of the Risks of Mixtures of Major Use Veterinary Antibiotics in European Surface Waters

Effects of single veterinary antibiotics on a range of aquatic organisms have been explored in many studies. In reality, surface waters will be exposed to mixtures of these substances. In this study, we present an approach for establishing risks of antibiotic mixtures to surface waters and illustrate this by assessing risks of mixtures of three major use antibiotics (trimethoprim, tylosin, and lincomycin) to algal and cyanobacterial species in European surface waters. Ecotoxicity tests were initially performed to assess the combined effects of the antibiotics to the cyanobacteria Anabaena flos-aquae. The results were used to evaluate two mixture prediction models: concentration addition (CA) and independent action (IA). The CA model performed best at predicting the toxicity of the mixture with the experimental 96 h EC50 for the antibiotic mixture being 0.248 μmol/L compared to the CA predicted EC50 of 0.21 μmol/L. The CA model was therefore used alongside predictions of exposure for different European scenarios and estimations of hazards obtained from species sensitivity distributions to estimate risks of mixtures of the three antibiotics. Risk quotients for the different scenarios ranged from 0.066 to 385 indicating that the combination of three substances could be causing adverse impacts on algal communities in European surface waters. This could have important implications for primary production and nutrient cycling. Tylosin contributed most to the risk followed by lincomycin and trimethoprim. While we have explored only three antibiotics, the combined experimental and modeling approach could readily be applied to the wider range of antibiotics that are in use

Understanding employee resourcing in construction organizations

In recent years the literature on employee resourcing has consistently advocated the importance of adopting a holistic, strategic approach to employee deployment decision making rather than adopting a reactive needs-based approach. This is particularly problematic in construction where the multi-project environment leads to constantly changing resource requirements and to changing demands over a project's life cycle. This can lead to inappropriate decisions, which fail to meet the longer-term needs of both construction organizations and their employees. A structured and comprehensive understanding of the current project team deployment practices within large construction organizations was developed. Project deployment practices were examined within seven case study contracting firms. The emergent themes that shaped the decision-making processes were grouped into five broad clusters comprising human resource planning, performance/career management, team deployment, employee involvement and training and development. The research confirms that a reactive and ad hoc approach to the function prevails within the firms investigated. This suggests a weak relationship between the deployment process and human resource planning, team deployment, performance management, employee involvement and training and development activities. It is suggested that strategic HR-business partnering could engender more transparent and productive relationships in this crucial area