Environmental aspects and challenges of oilseed produced biodiesel in Southeast Asia

Research on alternative fuel for the vehemently growing number of automotivesis intensified due to environmental reasons rather than turmoil in energy price and supply. From the policy and steps to emphasis the use of biofuel by governments all around the world, this can be comprehended that biofuel have placed itself as a number one substitute for fossil fuels. These phenomena made Southeast Asia a prominent exporter of biodiesel. But thrust in biodiesel production from oilseeds of palm and Jatropha curcas in Malaysia, Indonesia and Thailand is seriously threatening environmental harmony. This paper focuses on this critical issue of biodiesels environmental impacts, policy, standardization of this region as well as on the emission of biodiesel in automotive uses. To draw a bottom line on feasibilities of different feedstock of biodiesel, a critical analysis on oilseed yield rate, land use, engine emissions and oxidation stability is reviewed. Palm oil based biodiesel is clearly ahead in all these aspects of feasibility, except in the case of NOx where it lags from conventional petro diesel.

Test and evaluation of non conventional instrument transformers and sampled value process bus on Powerlink's transmission network

IEC 61850 Process Bus technology has the potential to improve cost, performance and reliability of substation design. Substantial costs associated with copper wiring (designing, documentation, construction, commissioning and troubleshooting) can be reduced with the application of digital Process Bus technology, especially those based upon international standards. An IEC 61850-9-2 based sampled value Process Bus is an enabling technology for the application of Non-Conventional Instrument Transformers (NCIT). Retaining the output of the NCIT in its native digital form, rather than conversion to an analogue output, allows for improved transient performance, dynamic range, safety, reliability and reduced cost. In this paper we report on a pilot installation using NCITs communicating across a switched Ethernet network using the UCAIug Implementation Guideline for IEC 61850-9-2 (9-2 Light Edition or 9-2LE). This system was commissioned in a 275 kV Line Reactor bay at Powerlink Queensland’s Braemar substation in 2009, with sampled value protection IEDs 'shadowing' the existing protection system. The results of commissioning tests and twelve months of service experience using a Fibre Optic Current Transformer (FOCT) from Smart Digital Optics (SDO) are presented, including the response of the system to fault conditions. A number of remaining issues to be resolved to enable wide-scale deployment of NCITs and IEC 61850-9-2 Process Bus technology are also discussed.

In vitro and in vivo examination of the cell surface glycoprotein CDCP1

A number of reports have demonstrated the importance of the CUB domaincontaining protein 1 (CDCP1) in facilitating cancer progression in animal models and the potential of this protein as a prognostic marker in several malignancies. CDCP1 facilitates metastasis formation in animal models by negatively regulating anoikis, a type of apoptosis triggered by the loss of attachment signalling from cell-cell contacts or cell-extra cellular matrix (ECM) contacts. Due to the important role CDCP1 plays in cancer progression in model systems, it is considered a potential drug target to prevent the metastatic spread of cancers. CDCP1 is a highly glycosylated 836 amino acid cell surface protein. It has structural features potentially facilitating protein-protein interactions including 14 N-glycosylation sites, three CUB-like domains, 20 cysteine residues likely to be involved in disulfide bond formation and five intracellular tyrosine residues. CDCP1 interacts with a variety of proteins including Src family kinases (SFKs) and protein kinase C ä (PKCä). Efforts to understand the mechanisms regulating these interactions have largely focussed on three CDCP1 tyrosine residues Y734, Y743 and Y762. CDCP1-Y734 is the site where SFKs phosphorylate and bind to CDCP1 and mediate subsequent phosphorylation of CDCP1-Y743 and -Y762 which leads to binding of PKCä at CDCP1-Y762. The resulting trimeric protein complex of SFK•CDCP1•PKCä has been proposed to mediate an anti-apoptotic cell phenotype in vitro, and to promote metastasis in vivo. The effect of mutation of the three tyrosines on interactions of CDCP1 with SFKs and PKCä and the consequences on cell phenotype in vitro and in vivo have not been examined. CDCP1 has a predicted molecular weight of ~90 kDa but is usually detected as a protein which migrates at ~135 kDa by Western blot analysis due to its high degree of glycosylation. A low molecular weight form of CDCP1 (LMWCDCP1) of ~70 kDa has been found in a variety of cancer cell lines. The mechanisms leading to the generation of LMW-CDCP1 in vivo are not well understood but an involvement of proteases in this process has been proposed. Serine proteases including plasmin and trypsin are able to proteolytically process CDCP1. In addition, the recombinant protease domain of the serine protease matriptase is also able to cleave the recombinant extracellular portion of CDCP1. Whether matriptase is able to proteolytically process CDCP1 on the cell surface has not been examined. Importantly, proteolytic processing of CDCP1 by trypsin leads to phosphorylation of its cell surface-retained portion which suggests that this event leads to initiation of an intracellular signalling cascade. This project aimed to further examine the biology of CDCP1 with a main of focus on exploring the roles played by CDCP1 tyrosine residues. To achieve this HeLa cells stably expressing CDCP1 or the CDCP1 tyrosine mutants Y734F, Y743F and Y762F were generated. These cell lines were used to examine: • The roles of the tyrosine residues Y734, Y743 and Y762 in mediating interactions of CDCP1 with binding proteins and to examine the effect of the stable expression on HeLa cell morphology. • The ability of the serine protease matriptase to proteolytically process cell surface CDCP1 and to examine the consequences of this event on HeLa cell phenotype and cell signalling in vitro. • The importance of these residues in processes associated with cancer progression in vitro including adhesion, proliferation and migration. • The role of these residues on metastatic phenotype in vivo and the ability of a function-blocking anti-CDCP1 antibody to inhibit metastasis in the chicken embryo chorioallantoic membrane (CAM) assay. Interestingly, biochemical experiments carried out in this study revealed that mutation of certain CDCP1 tyrosine residues impacts on interactions of this protein with binding proteins. For example, binding of SFKs as well as PKCä to CDCP1 was markedly decreased in HeLa-CDCP1-Y734F cells, and binding of PKCä was also reduced in HeLa-CDCP1-Y762F cells. In contrast, HeLa-CDCP1-Y743F cells did not display altered interactions with CDCP1 binding proteins. Importantly, observed differences in interactions of CDCP1 with binding partners impacted on basal phosphorylation of CDCP1. It was found that HeLa-CDCP1, HeLa-CDCP1-Y743F and -Y762F displayed strong basal levels of CDCP1 phosphorylation. In contrast, HeLa-CDCP1-Y734F cells did not display CDCP1 phosphorylation but exhibited constitutive phosphorylation of focal adhesion kinase (FAK) at tyrosine 861. Significantly, subsequent investigations to examine this observation suggested that CDCP1-Y734 and FAK-Y861 are competitive substrates for SFK-mediated phosphorylation. It appeared that SFK-mediated phosphorylation of CDCP1- Y734 and FAK-Y861 is an equilibrium which shifts depending on the level of CDCP1 expression in HeLa cells. This suggests that the level of CDCP1 expression may act as a regulatory mechanism allowing cells to switch from a FAK-Y861 mediated pathway to a CDCP1-Y734 mediated pathway. This is the first time that a link between SFKs, CDCP1 and FAK has been demonstrated. One of the most interesting observations from this work was that CDCP1 altered HeLa cell morphology causing an elongated and fibroblastic-like appearance. Importantly, this morphological change depended on CDCP1- Y734. In addition, it was observed that this change in cell morphology was accompanied by increased phosphorylation of SFK-Y416. This suggests that interactions of SFKs with CDCP1-Y734 increases SFK activity since SFKY416 is critical in regulating kinase activity of these proteins. The essential role of SFKs in mediating CDCP1-induced HeLa cell morphological changes was demonstrated using the SFK-selective inhibitor SU6656. This inhibitor caused reversion of HeLa-CDCP1 cell morphology to an epithelial appearance characteristic of HeLa-vector cells. Significantly, in vitro studies revealed that certain CDCP1-mediated cell phenotypes are mediated by cellular pathways dependent on CDCP1 tyrosine residues whereas others are independent of these sites. For example, CDCP1 expression caused a marked increase in HeLa cell motility that was independent of CDCP1 tyrosine residues. In contrast, CDCP1- induced decrease in HeLa cell proliferation was most prominent in HeLa- CDCP1-Y762F cells, potentially indicating a role for this site in regulating proliferation in HeLa cells. Another cellular event which was identified to require phosphorylation of a particular CDCP1 tyrosine residue is adhesion to fibronectin. It was observed that the CDCP1-mediated strong decrease in adhesion to fibronectin is mostly restored in HeLa-CDCP1-Y743F cells. This suggests a possible role for CDCP1-Y743 in causing a CDCP1-mediated decrease in adhesion. Data from in vivo experiments indicated that HeLa-CDCP1-Y734F cells are more metastic than HeLa-CDCP1 cells in vivo. This indicates that interaction of CDCP1 with SFKs and PKCä may not be required for CDCP1-mediated metastasis formation of HeLa cells in vivo. The metastatic phenotype of these cells may be caused by signalling involving FAK since HeLa-CDCP1- Y734F cells are the only CDCP1 expressing cells displaying constitutive phosphorylation of FAK-Y861. HeLa-CDCP1-Y762F cells displayed a very low metastatic ability which suggests that this CDCP1 tyrosine residue is important in mediating a pro-metastatic phenotype in HeLa cells. More detailed exploration of cellular events occurring downstream of CDCP1-Y734 and -Y762 may provide important insights into the mechanisms altering the metastatic ability of CDCP1 expressing HeLa cells. Complementing the in vivo studies, anti-CDCP1 antibodies were employed to assess whether these antibodies are able to inhibit metastasis of CDCP1 and CDCP1 tyrosine mutants expressing HeLa cells. It was found that HeLa- CDCP1-Y734F cells were the only cell line which was markedly reduced in the ability to metastasise. In contrast, the ability of HeLa-CDCP1, HeLa- CDCP1-Y743F and -Y762F cells to metastasise in vivo was not inhibited. These data suggest a possible role of interactions of CDCP1 with SFKs, occurring at CDCP1-Y734, in preventing an anti-metastatic effect of anti- CDCP1 antibodies in vivo. The proposal that SFKs may play a role in regulating anti-metastatic effects of anti-CDCP1 antibodies was supported by another experiment where differences between HeLa-CDCP1 cells and CDCP1 expressing HeLa cells (HeLa-CDCP1-S) from collaborators at the Scripps Research Institute were examined. It was found that HeLa-CDCP1-S cells express different SFKs than CDCP1 expressing HeLa cells generated for this study. This is important since HeLa-CDCP1-S cells can be inhibited in their metastatic ability using anti-CDCP1 antibodies in vivo. Importantly, these data suggest that further examinations of the roles of SFKs in facilitating anti-metastatic effects of anti-CDCP1 antibodies may give insights into how CDCP1 can be blocked to prevent metastasis in vivo. This project also explored the ability of the serine protease matriptase to proteolytically process cell surface localised CDCP1 because it is unknown whether matriptase can cleave cell surface CDCP1 as it has been reported for other proteases such as trypsin and plasmin. Furthermore, the consequences of matriptase-mediated proteolysis on cell phenotype in vitro and cell signalling were examined since recent reports suggested that proteolysis of CDCP1 leads to its phosphorylation and may initiate cell signalling and consequently alter cell phenotype. It was found that matriptase is able to proteolytically process cell surface CDCP1 at low nanomolar concentrations which suggests that cleavage of CDCP1 by matriptase may facilitate the generation of LWM-CDCP1 in vivo. To examine whether matriptase-mediated proteolysis induced cell signalling anti-phospho Erk 1/2 Western blot analysis was performed as this pathway has previously been examined to study signalling in response to proteolytic processing of cell surface proteins. It was found that matriptase-mediated proteolysis in CDCP1 expressing HeLa cells initiated intracellular signalling via Erk 1/2. Interestingly, this increase in phosphorylation of Erk 1/2 was also observed in HeLa-vector cells. This suggested that initiation of cell signalling via Erk 1/2 phosphorylation as a result of matriptase-mediated proteolysis occurs by pathways independent of CDCP1. Subsequent investigations measuring the flux of free calcium ions and by using a protease-activated receptor 2 (PAR2) agonist peptide confirmed this hypothesis. These data suggested that matriptase-mediated proteolysis results in cell signalling via a pathway induced by the activation of PAR2 rather than by CDCP1. This indicates that induction of cell signalling in HeLa cells as a consequence of matriptase-mediated proteolysis occurs via signalling pathways which do not involve phosphorylation of Erk 1/2. Consequently, it appears that future attempts should focus on the examination of cellular pathways other than Erk 1/2 to elucidate cell signalling initiated by matriptase-mediated proteolytic processing of CDCP1. The data presented in this thesis has explored in vitro and in vivo aspects of the biology of CDCP1. The observations summarised above will permit the design of future studies to more precisely determine the role of CDCP1 and its binding partners in processes relevant to cancer progression. This may contribute to further defining CDCP1 as a target for cancer treatment.

Technology strategy for re-engineering design and construction

Automation technology can provide construction firms with a number of competitive advantages. Technology strategy guides a firm's approach to all technology, including automation. Engineering management educators, researchers, and construction industry professionals need improved understanding of how technology affects results, and how to better target investments to improve competitive performance. A more formal approach to the concept of technology strategy can benefit the construction manager in his efforts to remain competitive in increasingly hostile markets. This paper recommends consideration of five specific dimensions of technology strategy within the overall parameters of market conditions, firm capabilities and goals, and stage of technology evolution. Examples of the application of this framework in the formulation of technology strategy are provided for CAD applications, co-ordinated positioning technology and advanced falsework and formwork mechanisation to support construction field operations. Results from this continuing line of research can assist managers in making complex and difficult decisions regarding reengineering construction processes in using new construction technology and benefit future researchers by providing new tools for analysis. Through managing technology to best suit the existing capabilities of their firm, and addressing the market forces, engineering managers can better face the increasingly competitive environment in which they operate.

Natural balancing of flying capacitor voltages in multicell inverter under PD carrier-based PWM

The flying capacitor multicell inverter (FCMI) possesses natural balancing property. With the phase-shifted (PS) carrier-based scheme, natural balancing can be achieved in a straightforward manner. However, to achieve natural balancing with the harmonically optimal phase-disposition (PD) carrierbased scheme, the conventional approaches require (n-1) x (n-1) trapezoidal carrier signals for an n-level inverter, which is (n-1) x (n-2) times more than that in the standard PD scheme. This paper proposes two improved natural balancing strategies for FMI under PD scheme, which use the same (n-1) carrier signals as used in the standard PD scheme. In the first scheme, on-line detection is performed of the band in which the modulation signal is located, corresponding period number of the carrier, and rising or falling half cycle of the carrier waveform to generate the switching signals based on certain rules. In the second strategy, the output voltage level selection is first processed and the switching signals are then generated according to a rule based on preferential cell selection algorithm. These methods are easy to use and can be simply implemented as compared to the other available methods. Simulation and experimental results are presented for a five-level inverter to verify these proposed schemes.

A biologically inspired object spectral-texture descriptor and its application to vegetation classification in power-line corridors

The use of appropriate features to represent an output class or object is critical for all classification problems. In this paper, we propose a biologically inspired object descriptor to represent the spectral-texture patterns of image-objects. The proposed feature descriptor is generated from the pulse spectral frequencies (PSF) of a pulse coupled neural network (PCNN), which is invariant to rotation, translation and small scale changes. The proposed method is first evaluated in a rotation and scale invariant texture classification using USC-SIPI texture database. It is further evaluated in an application of vegetation species classification in power line corridor monitoring using airborne multi-spectral aerial imagery. The results from the two experiments demonstrate that the PSF feature is effective to represent spectral-texture patterns of objects and it shows better results than classic color histogram and texture features.

The Kinaesthetic Fusion Effect : mechanisms and extensions

This study investigated the Kinaesthetic Fusion Effect (KFE) that was first described by Craske and Kenny in 1981. It was reported that when, without vision, participants pressed a button that resulted in a probe simultaneously touching the contralateral limb at a displaced location, they perceived an apparent change in limb length. The current study did not fully replicate these earlier findings. Participants did not perceive any reduction in the sagittal separation of the button and probe following repeated exposure to the tactile stimuli that was present on both arms. However, a localised and partial medio-lateral fusion was observed, with the touched positions seeming closer together. In addition, tactile acuity was found to decrease progressively for distal positions of the upper limb and a foreshortening effect was found which may result from a line-of-sight judgment and represent a feature of the reporting method used. A number of years have elapsed since the description of the original KFE. Although frequently cited in the literature, there has been no further investigation into the mechanisms of action. The results of the current study are considered in light of more recent literature concerning intersensory integration. Future research should focus on further clarification for the specific conditions that must be present for a fusion effect to occur. Finally, this thesis will benefit future studies that require participants to report the perceived locations of the unseen limbs.

Innovation, interaction, and inclusion : heritage luxury brands in collusion with the consumer

Many luxury heritage brands operate on the misconception that heritage is interchangeable with history rather than representative of the emotional response they originally developed in their customer. This idea of heritage as static history inhibits innovation, prevents dynamic renewal and impedes their ability to redefine, strengthen and position their brand in current and emerging marketplaces. This paper examines a number of heritage luxury brands that have successfully identified the original emotional responses they developed in their customers and, through innovative approaches in design, marketing, branding and distribution evoke these responses in contemporary consumers. Using heritage and innovation hand-in-hand, these brands have continued to grow and develop a vision of heritage that incorporates both historical and contemporary ideas to meet emerging customer needs. While what constitutes a ‘luxury’ item is constantly challenged in this era of accessible luxury products, up-scaling and aspirational spending, this paper sees consumers’ emotional needs as the key element in defining the concept of luxury. These emotional qualities consistently remain relevant due to their ability to enhance a positive sense of identity for the brand user. Luxury is about the ‘experience’ not just the product providing the consumer with a sense of enhanced status or identity through invoked feelings of exclusivity, authenticity, quality, uniqueness and culture. This paper will analyse luxury heritage brands that have successfully combined these emotional values with those of their ‘heritage’ to create an aura of authenticity and nostalgia that appeals to contemporary consumers. Like luxury, the line where clothing becomes fashion is blurred in the contemporary fashion industry; however, consumer emotion again plays an important role. For example, clothing becomes ‘fashion’ for consumers when it affects their self perception rather than fulfilling basic functions of shelter and protection. Successful luxury heritage brands can enhance consumers’ sense of self by involving them in the ‘experience’ and ‘personality’ of the brand so they see it as a reflection of their own exclusiveness, authentic uniqueness, belonging and cultural value. Innovation is a valuable tool for heritage luxury brands to successfully generate these desired emotional responses and meet the evolving needs of contemporary consumers. While traditionally fashion has been a monologue from brand to consumer, new technology has given consumers a voice to engage brands in a conversation to express their evolving needs, ideas and feedback. As a result, in this consumer-empowered era of information sharing, this paper defines innovation as the ability of heritage luxury brands to develop new design and branding strategies in response to this consumer feedback while retaining the emotional core values of their heritage. This paper analyses how luxury heritage brands can effectively position themselves in the contemporary marketplace by separating heritage from history to incorporate innovative strategies that will appeal to consumer needs of today and tomorrow.

Design of compact coupled-line couplers

The size of rat-race and branch-line couplers can be reduced by using periodic loading or artificial transmission lines. The objective of this work is to extend the idea of size reduction through periodic loading to coupled-line 90° hybrids. A procedure for the extraction of the characteristic parameters of a coupled-line 4-port from a single set of S-parameters is described. This method can be employed to design of coupled artificial transmission line couplers of arbitrary geometry. The procedure is illustrated through the design a broadside-coupled stripline hybrid, periodically loaded with stubs. Measured results for a prototype coupler confirm the validity of the theory.

Innovation, interaction, and inclusion : heritage luxury brands in collusion with the consumer

Many luxury heritage brands operate on the misconception that heritage is interchangeable with history rather than representative of the emotional response they originally developed in their customer. This idea of heritage as static history inhibits innovation, prevents dynamic renewal and impedes their ability to redefine, strengthen and position their brand in current and emerging marketplaces. This paper examines a number of heritage luxury brands that have successfully identified the original emotional responses they developed in their customers and, through innovative approaches in design, marketing, branding and distribution evoke these responses in contemporary consumers. Using heritage and innovation hand-in-hand, these brands have continued to grow and develop a vision of heritage that incorporates both historical and contemporary ideas to meet emerging customer needs. While what constitutes a ‘luxury’ item is constantly challenged in this era of accessible luxury products, up scaling and aspirational spending, this paper sees consumers’ emotional needs as the key element in defining the concept of luxury. These emotional qualities consistently remain relevant due to their ability to enhance a positive sense of identity for the brand user. Luxury is about the ‘experience’ not just the product providing the consumer with a sense of enhanced status or identity through invoked feelings of exclusivity, authenticity, quality, uniqueness and culture. This paper will analyse luxury heritage brands that have successfully combined these emotional values with those of their ‘heritage’ to create an aura of authenticity and nostalgia that appeals to contemporary consumers. Like luxury, the line where clothing becomes fashion is blurred in the contemporary fashion industry; however, consumer emotion again plays an important role. For example, clothing becomes ‘fashion’ for consumers when it affects their self perception rather than fulfilling basic functions of shelter and protection. Successful luxury heritage brands can enhance consumers’ sense of self by involving them in the ‘experience’ and ‘personality’ of the brand so they see it as a reflection of their own exclusiveness, authentic uniqueness, belonging and cultural value. Innovation is a valuable tool for heritage luxury brands to successfully generate these desired emotional responses and meet the evolving needs of contemporary consumers. While traditionally fashion has been a monologue from brand to consumer, new technology has given consumers a voice to engage brands in a conversation to express their evolving needs, ideas and feedback. As a result, in this consumer-empowered era of information sharing, this paper defines innovation as the ability of heritage luxury brands to develop new design and branding strategies in response to this consumer feedback while retaining the emotional core values of their heritage. This paper analyses how luxury heritage brands can effectively position themselves in the contemporary marketplace by separating heritage from history to incorporate innovative strategies that will appeal to consumer needs of today and tomorrow.

Measurement of total body water (TBW) and total energy expenditure (TEE) using stable isotopes

Understanding the relationship between diet, physical activity and health in humans requires accurate measurement of body composition and daily energy expenditure. Stable isotopes provide a means of measuring total body water and daily energy expenditure under free-living conditions. While the use of isotope ratio mass spectrometry (IRMS) for the analysis of 2H (Deuterium) and 18O (Oxygen-18) is well established in the field of human energy metabolism research, numerous questions remain regarding the factors which influence analytical and measurement error using this methodology. This thesis was comprised of four studies with the following emphases. The aim of Study 1 was to determine the analytical and measurement error of the IRMS with regard to sample handling under certain conditions. Study 2 involved the comparison of TEE (Total daily energy expenditure) using two commonly employed equations. Further, saliva and urine samples, collected at different times, were used to determine if clinically significant differences would occur. Study 3 was undertaken to determine the appropriate collection times for TBW estimates and derived body composition values. Finally, Study 4, a single case study to investigate if TEE measures are affected when the human condition changes due to altered exercise and water intake. The aim of Study 1 was to validate laboratory approaches to measure isotopic enrichment to ensure accurate (to international standards), precise (reproducibility of three replicate samples) and linear (isotope ratio was constant over the expected concentration range) results. This established the machine variability for the IRMS equipment in use at Queensland University for both TBW and TEE. Using either 0.4mL or 0.5mL sample volumes for both oxygen-18 and deuterium were statistically acceptable (p>0.05) and showed a within analytical variance of 5.8 Delta VSOW units for deuterium, 0.41 Delta VSOW units for oxygen-18. This variance was used as “within analytical noise” to determine sample deviations. It was also found that there was no influence of equilibration time on oxygen-18 or deuterium values when comparing the minimum (oxygen-18: 24hr; deuterium: 3 days) and maximum (oxygen-18: and deuterium: 14 days) equilibration times. With regard to preparation using the vacuum line, any order of preparation is suitable as the TEE values fall within 8% of each other regardless of preparation order. An 8% variation is acceptable for the TEE values due to biological and technical errors (Schoeller, 1988). However, for the automated line, deuterium must be assessed first followed by oxygen-18 as the automated machine line does not evacuate tubes but merely refills them with an injection of gas for a predetermined time. Any fractionation (which may occur for both isotopes), would cause a slight elevation in the values and hence a lower TEE. The purpose of the second and third study was to investigate the use of IRMS to measure the TEE and TBW of and to validate the current IRMS practices in use with regard to sample collection times of urine and saliva, the use of two TEE equations from different research centers and the body composition values derived from these TEE and TBW values. Following the collection of a fasting baseline urine and saliva sample, 10 people (8 women, 2 men) were dosed with a doubly labeled water does comprised of 1.25g 10% oxygen-18 and 0.1 g 100% deuterium/kg body weight. The samples were collected hourly for 12 hrs on the first day and then morning, midday, and evening samples were collected for the next 14 days. The samples were analyzed using an isotope ratio mass spectrometer. For the TBW, time to equilibration was determined using three commonly employed data analysis approaches. Isotopic equilibration was reached in 90% of the sample by hour 6, and in 100% of the sample by hour 7. With regard to the TBW estimations, the optimal time for urine collection was found to be between hours 4 and 10 as to where there was no significant difference between values. In contrast, statistically significant differences in TBW estimations were found between hours 1-3 and from 11-12 when compared with hours 4-10. Most of the individuals in this study were in equilibrium after 7 hours. The TEE equations of Prof Dale Scholler (Chicago, USA, IAEA) and Prof K.Westerterp were compared with that of Prof. Andrew Coward (Dunn Nutrition Centre). When comparing values derived from samples collected in the morning and evening there was no effect of time or equation on resulting TEE values. The fourth study was a pilot study (n=1) to test the variability in TEE as a result of manipulations in fluid consumption and level of physical activity; the magnitude of change which may be expected in a sedentary adult. Physical activity levels were manipulated by increasing the number of steps per day to mimic the increases that may result when a sedentary individual commences an activity program. The study was comprised of three sub-studies completed on the same individual over a period of 8 months. There were no significant changes in TBW across all studies, even though the elimination rates changed with the supplemented water intake and additional physical activity. The extra activity may not have sufficiently strenuous enough and the water intake high enough to cause a significant change in the TBW and hence the CO2 production and TEE values. The TEE values measured show good agreement based on the estimated values calculated on an RMR of 1455 kcal/day, a DIT of 10% of TEE and activity based on measured steps. The covariance values tracked when plotting the residuals were found to be representative of “well-behaved” data and are indicative of the analytical accuracy. The ratio and product plots were found to reflect the water turnover and CO2 production and thus could, with further investigation, be employed to identify the changes in physical activity.