Development of an in-vitro model system to investigate the mechanism of muscle protein catabolism induced by proteolysis-inducing factor

The mechanism of muscle protein catabolism induced by proteolysis-inducing factor, produced by cachexia-inducing murine and human tumours has been studied in vitro using C2C12 myoblasts and myotubes. In both myoblasts and myotubes protein degradation was enhanced by proteolysis-inducing factor after 24 h incubation. In myoblasts this followed a bell-shaped dose-response curve with maximal effects at a proteolysis-inducing factor concentration between 2 and 4 nM, while in myotubes increased protein degradation was seen at all concentrations of proteolysis-inducing factor up to 10 nM, again with a maximum of 4 nM proteolysis-inducing factor. Protein degradation induced by proteolysis-inducing factor was completely attenuated in the presence of cycloheximide (1 μM), suggesting a requirement for new protein synthesis. In both myoblasts and myotubes protein degradation was accompanied by an increased expression of the α-type subunits of the 20S proteasome as well as functional activity of the proteasome, as determined by the 'chymotrypsin-like' enzyme activity. There was also an increased expression of the 19S regulatory complex as well as the ubiquitin-conjugating enzyme (E214k), and in myotubes a decrease in myosin expression was seen with increasing concentrations of proteolysis-inducing factor. These results show that proteolysis-inducing factor co-ordinately upregulates both ubiquitin conjugation and proteasome activity in both myoblasts and myotubes and may play an important role in the muscle wasting seen in cancer cachexia. © 2002 Cancer Research UK.

Complementary methods to investigate the development of clogging within a horizontal sub-surface flow tertiary treatment wetland

A combination of experimental methods was applied at a clogged, horizontal subsurface flow (HSSF) municipal wastewater tertiary treatment wetland (TW) in the UK, to quantify the extent of surface and subsurface clogging which had resulted in undesirable surface flow. The three dimensional hydraulic conductivity profile was determined, using a purpose made device which recreates the constant head permeameter test in-situ. The hydrodynamic pathways were investigated by performing dye tracing tests with Rhodamine WT and a novel multi-channel, data-logging, flow through Fluorimeter which allows synchronous measurements to be taken from a matrix of sampling points. Hydraulic conductivity varied in all planes, with the lowest measurement of 0.1 md1 corresponding to the surface layer at the inlet, and the maximum measurement of 1550 md1 located at a 0.4m depth at the outlet. According to dye tracing results, the region where the overland flow ceased received five times the average flow, which then vertically short-circuited below the rhizosphere. The tracer break-through curve obtained from the outlet showed that this preferential flow-path accounted for approximately 80% of the flow overall and arrived 8 h before a distinctly separate secondary flow-path. The overall volumetric efficiencyof the clogged system was 71% and the hydrology was simulated using a dual-path, dead-zone storage model. It is concluded that uneven inlet distribution, continuous surface loading and high rhizosphere resistance is responsible for the clog formation observed in this system. The average inlet hydraulic conductivity was 2 md1, suggesting that current European design guidelines, which predict that the system will reach an equilibrium hydraulic conductivity of 86 md1, do not adequately describe the hydrology of mature systems.

An X-ray micro-fluorescence study to investigate the distribution of Al, Si, P and Ca ions in the surrounding soft tissue after implantation of a calcium phosphate-mullite ceramic composite in a rabbit animal model

Synthetic calcium phosphates, despite their bioactivity, are brittle. Calcium phosphate-mullite composites have been suggested as potential dental and bone replacement materials which exhibit increased toughness. Aluminium, present in mullite, has however been linked to bone demineralisation and neurotoxicity: it is therefore important to characterise the materials fully in order to understand their in vivo behaviour. The present work reports the compositional mapping of the interfacial region of a calcium phosphate-20 wt% mullite biocomposite/soft tissue interface, obtained from the samples implanted into the long bones of healthy rabbits according to standard protocols (ISO-10993) for up to 12 weeks. X-ray micro-fluorescence was used to map simultaneously the distribution of Al, P, Si and Ca across the ceramic-soft tissue interface. A well defined and sharp interface region was present between the ceramic and the surrounding soft tissue for each time period examined. The concentration of Al in the surrounding tissue was found to fall by two orders of magnitude, to the background level, within similar to 35 mu m of the implanted ceramic.

A Conceptual Foresight Model to Investigate the Adoption of Radio Frequency Identification Technology in the English National Health Service

Radio Frequency Identification Technology (RFID) adoption in healthcare settings has the potential to reduce errors, improve patient safety, streamline operational processes and enable the sharing of information throughout supply chains. RFID adoption in the English NHS is limited to isolated pilot studies. Firstly, this study investigates the drivers and inhibitors to RFID adoption in the English NHS from the perspective of the GS1 Healthcare User Group (HUG) tasked with coordinating adoption across private and public sectors. Secondly a conceptual model has been developed and deployed, combining two of foresight’s most popular methods; scenario planning and technology roadmapping. The model addresses the weaknesses of each foresight technique as well as capitalizing on their individual, inherent strengths. Semi structured interviews, scenario planning workshops and a technology roadmapping exercise were conducted with the members of the HUG over an 18-month period. An action research mode of enquiry was utilized with a thematic analysis approach for the identification and discussion of the drivers and inhibitors of RFID adoption. The results of the conceptual model are analysed in comparison to other similar models. There are implications for managers responsible for RFID adoption in both the NHS and its commercial partners, and for foresight practitioners. Managers can leverage the insights gained from identifying the drivers and inhibitors to RFID adoption by making efforts to influence the removal of inhibitors and supporting the continuation of the drivers. The academic contribution of this aspect of the thesis is in the field of RFID adoption in healthcare settings. Drivers and inhibitors to RFID adoption in the English NHS are compared to those found in other settings. The implication for technology foresight practitioners is a proof of concept of a model combining scenario planning and technology roadmapping using a novel process. The academic contribution to the field of technology foresight is the conceptual development of foresight model that combines two popular techniques and then a deployment of the conceptual foresight model in a healthcare setting exploring the future of RFID technology.

Using simulation to assess the introduction of technology in a continuous operations process

Purpose – To investigate the role of simulation in the introduction of technology in a continuous operations process. Design/methodology/approach – A case-based research method was chosen with the aim to provide an exemplar of practice and test the proposition that the use of simulation can improve the implementation and running of conveyor systems in continuous process facilities. Findings – The research determines the optimum rate of re-introduction of inventory to a conveyor system generated during a breakdown event. Research limitations/implications – More case studies are required demonstrating the operational and strategic benefits that can be gained by using simulation to assess technology in organisations. Practical implications – A practical outcome of the study was the implementation of a policy for the manual re-introduction of inventory on a conveyor line after a breakdown event had occurred. Originality/value – The paper presents a novel example of the use of simulation to estimate the re-introduction rate of inventory after a breakdown event on a conveyor line. The paper highlights how by addressing this operational issue, ahead of implementation, the likelihood of the success of the strategic decision to acquire the technology can be improved.

Signaling pathways initiated by beta-hydroxy-beta-methylbutyrate to attenuate the depression of protein synthesis in skeletal muscle in response to cachectic stimuli

To investigate the mechanism by which beta-hydroxy-beta-methylbutyrate (HMB) attenuates the depression of protein synthesis in the skeletal muscle of cachectic mice, a study has been carried out in murine myotubes in the presence of proteolysis-inducing factor (PIF). PIF inhibited protein synthesis by 50% within 4 h, and this was effectively attenuated by HMB (25-50 muM). HMB (50 muM) alone stimulated protein synthesis, and this was attenuated by rapamycin (27 nM), an inhibitor of mammalian target of rapamycin (mTOR). Further evidence for an involvement of this pathway was shown by an increased phosphorylation of mTOR, the 70-kDa ribosomal S6 kinase (p70(S6k)), and initiation factor 4E-binding protein (4E-BP1) and an increased association of eukaryotic initiation factor 2 (eIF4E) with eIF4G. PIF alone induced a transient (1-2 h) stimulation of phosphorylation of mTOR and p70(S6k). However, in the presence of HMB, phosphorylation of mTOR, p70(S6k), and 4E-BP1 was increased, and inactive 4E-BP1-eIF4E complex was reduced, whereas the active eIF4G.eIF4E complex was increased, suggesting continual stimulation of protein synthesis. HMB alone reduced phosphorylation of elongation factor 2, but this effect was not seen in the presence of PIF. PIF induced autophosphorylation of the double-strand RNA-dependent protein kinase (PKR), leading to phosphorylation of eIF2 on the alpha-subunit, which would inhibit protein synthesis. However, in the presence of HMB, phosphorylation of PKR and eIF2alpha was attenuated, and this was also observed in skeletal muscle of cachectic mice administered HMB (0.25 g/kg). These results suggest that HMB attenuates the depression of protein synthesis by PIF in myotubes through multiple mechanisms.

An investigation to determine the influence of illumination sources on colour discrimination in normal subjects and smokers

Accurate colour vision testing requires using the correct illumination. With the plethora of 'daylight' lamps available, is there a cost-effective alternative to the discontinued MacBeth Easel lamp? Smoking is a known risk factor for macula degeneration. As the macula is responsible for colour discrimination, any toxin that affects it has the potential to influence colour discrimination. Aims: To find a costeffective light source for colour vision testing. To investigate the effect of smoking on colour discrimination. To explore how deuteranomalous trichromats compare with normal trichromats. Methods: Using the Ishihara colour vision test subjects were classified into the groups: 'Normal/Control', 'Smoker/Test', and 'Case Study' (subjects who failed the screening test and did not smoke). They completed the Farnsworth Munsell 100 Hue test under each of the three light sources: Phillips EcoHalo Twist (tungsten halogen - THL), Kosnic KCF07ALU/GU10-865 (compact fluorescent- CFL), and Deal Guardian Ltd. GU103X2WA4B-60 (light-emitting diode - LED) Results: 42 subjects took part in the study: 18 in the Normal/Control group, 18 in the Smoker/Test group, and 6 in the Case Study group. For the Normal/Control group the total error scores (TESs) were significantly lower with the CFL than with the THL (p = 0.017) as it was for the Case Study group (p = 0.009). No significant differences were found between the Normal/Control group and the Smoker/Test group for each light source. Decision tree analysis found pack years to be a significant variable for TES. Discussion: All three light sources were comparable with previous studies. The CFL provided better colour discrimination than the LED despite them both being 6500 K. Deuteranomalous trichromats showed a greatest deviation than normal trichromats using the LED. Conclusions: The Kosnic KCF07ALU/GU10-865 is a cost-effective alternative for colour vision testing. Smoking appears to have an effect on colour vision, but requires further investigation.

Using an in vitro human airway model to study the toxic effects of components of e-cigarettes

Cigarette smoke is a complex mixture of more than 4000 hazardous chemicals including the carcinogenic benzopyrenes. Nicotine, the most potent component of tobacco, is responsible for the addictive nature of cigarettes and is a major component of e-cigarette cartridges. Our study aims to investigate the toxicity of nicotine with special emphasis on the replacement of animals. Furthermore, we intend to study the effect of nicotine, cigarette smoke and e-cigarette vapours on human airways. In our current work, the BEAS 2B human bronchial epithelial cell line was used to analyse the effect of nicotine in isolation, on cell viability. Concentrations of nicotine from 1.1µM to 75µM were added to 5x105 cells per well in a 96 well plate and incubated for 24 hours. Cell titre blue results showed that all the nicotine treated cells were more metabolically active than the control wells (cells alone). These data indicate that, under these conditions, nicotine does not affect cell viability and in fact, suggests that there is a stimulatory effect of nicotine on metabolism. We are now furthering this finding by investigating the pro-inflammatory response of these cells to nicotine by measuring cytokine secretion via ELISA. Further work includes analysing nicotine exposure at different time points and on other epithelial cells lines like Calu-3.

Barriers to capturing the value of advanced services and digitisation in the road transport industry

Servitization and digitization together provide significant opportunities to raise the performance and profitability of the road transport industry. To date these opportunities are only sparsely captured and the potential economic, social and environmental value is forgone. We set out to investigate the barriers to capturing the value of servitization and digitization in the road transport industry.

Analysis of organisational climate to improve the adequacy of primary health care in the State of Kuwait

This study covers two areas of contribution to the knowledge, firstly it tried to investigate rigourously the relationships of a number of factors believed that they may affect the climate perception, classified into three types to arrive to prove a hypothesis of the important role that qualification and personal factors play in shaping the climate perception, this is in contrast with situational factors. Secondly, the study tries to recluster the items of a wide-range applied scale for the measurement of climate named HAY in order to overcome the cross-cultural differences between the Kuwaiti and the American society, and to achieve a modified dimensions of climate for a civil service organisation in Kuwait. Furthermore, the study attempts to carry out a diagnostic test for the climate of the Ministry of Public Health in Kuwait, aiming to diagnose the perceived characteristics of the MoPH organisation, and suggests a number of areas to be given attention if an improvement is to be introduced. The study used extensively the statistical and the computer facilities to make the analysis more representing the field data, on the other hand this study is characterised by the very highly responsive rate of the main survey which would affect the findings reliability. Three main field studies are included, the first one was to conduct the main questionnaire where the second was to measure the "should be" climate by the experts of MoPH using the DELPHI technique, and the third was to conduct an extensive meeting with the very top management team in MoPH. Results of the first stage were subject to CLUSTER analysis for the reconstruction of the HAY tool, whereas comparative analysis was carried on between the results of the second and third stages on one side, the first from the other.