Transitioning from electrodialysis to reverse electrodialysis stack design for energy generation from high concentration salinity gradients.

In this study, stack design for high concentration gradient reverse electrodialysis operating in recycle is addressed. High concentration gradients introduce complex transport phenomena, which are exacerbated when recycling feeds; a strategy employed to improve system level energy efficiency. This unique challenge indicates that membrane properties and spacer thickness requirements may differ considerably from reverse electrodialysis for lower concentration gradients (e.g. seawater/river water), drawing closer parallels to electrodialysis stack design. Consequently, commercially available electrodialysis and reverse electrodialysis stack design was first compared for power generation from high concentration gradients. Higher gross power densities were identified for the reverse electrodialysis stack, due to the use of thinner membranes characterised by a higher permselectivity, which improved current. However, energy efficiency of the electrodialysis stack was twice that recorded for the reverse electrodialysis stack at low current densities, which was attributed to: (i) an increased residence time provided by the larger intermembrane distance, and (ii) reduced exergy losses of the electrodialysis membranes, which provided comparatively lower water permeance. Further in-depth investigation into membrane properties and spacer thickness identified that membranes characterised by an intermediate water permeability and ohmic resistance provided the highest power density and energy efficiency (Neosepta ACS/CMS), while wider intermembrane distances up to 0.3 mm improved energy efficiency. This study confirms that reverse electrodialysis stacks for high concentration gradients in recycle therefore demand design more comparable to electrodialysis stacks to drive energy efficiency, but when selecting membrane properties, the trade-off with permselectivity must also be considered to ensure economic viability.

Scale-up of reverse electrodialysis for energy generation from high concentration salinity gradients.

Whilst reverse electrodialysis (RED) has been extensively characterised for saline gradient energy from seawater/river water (0.5 M/0.02 M), less is known about RED stack design for high concentration salinity gradients (4 M/0.02 M), important to closed loop applications (e.g. thermal-to-electrical, energy storage). This study therefore focuses on the scale-up of RED stacks for high concentration salinity gradients. Higher velocities were required to attain a maximum Open Circuit Voltage (OCV) for 4 M/0.02 M, which gives a measure of the electrochemical potential of the cell. The experimental OCV was also much below the theoretical OCV, due to the greater boundary layer resistance observed, which is distinct from 0.5 M/0.02 M. However, negative net power density (net produced electrical power divided by total membrane area) was demonstrated with 0.5 M/0.02 M for larger stacks using shorter residence times (three stack sizes tested: 10 × 10cm, 10 × 20cm and 10 × 40cm). In contrast, the highest net power density was observed at the shortest residence time for the 4 M/0.02 M concentration gradient, as the increased ionic flux compensated for the pressure drop. Whilst comparable net power densities were determined for the 10 × 10cm and 10 × 40cm stacks using the 4 M/0.02 M concentration gradient, the osmotic and ionic transport mechanisms are distinct. Increasing cell pair number improved maximum current density. This subsequently increased power density, due to the reduction in boundary layer resistance, and may therefore be used to improve thermodynamic efficiency and power density from RED for high concentrations. Although comparable power densities may be achieved for small and large stacks, large stacks maybe preferred for high concentration salinity gradients due to the comparative benefit in thermodynamic efficiency in single pass. The greater current achieved by large stacks may also be complemented by an increase in cell pair number and current density optimisation to increase power density and reduce exergy losses.

Membrane distillation for concentrated blackwater: Influence of configuration (air gap, direct contact, vacuum) on selectivity and water productivity.

Water recovery from concentrated blackwater has been studied using air gap (AGMD), direct contact (DCMD) and vacuum membrane distillation (VMD) to deliver decentralised sanitation. Whilst good water quality was achieved with each configuration, differences in the rejection of volatile compounds was observed. VMD exhibited the highest rejection of volatiles, specifically ammoniacal nitrogen, of all the configurations but fouling inhibited total flux. DCMD exhibited a temperature dependent volatile rejection which resulted in poor rejection at lower feed temperatures (≤40 °C). AGMD was identified as the most promising configuration for application within decentralised sanitation, since the rejection of volatiles was consistent over a range of operating temperatures with ammonia rejection directly related to solution pH. An increase in organic colloids and particles due to faecal contamination reduced COD removal due to the induction of wetting, but was shown to be offset by adoption of a smaller pore size (0.1 µm), and when complemented with upstream solid-liquid separation within a fully integrated system, will provide a robust sanitation solution. Importantly, this work has shown that AGMD can recover water from concentrated blackwater close to international discharge and reuse regulations in a single stage process; this is significant as blackwater consists of only urine and faeces, and is thus 40 times more concentrated than municipal sewage. It is proposed that the water quality produced reflects a step change to delivering safe sanitation, and is complemented by a simple method for heat recovery integration this is similarly advantageous for resource constrained environments common to decentralised sanitation solutions.

The impact of hydraulic retention time on the performance of two configurations of anaerobic pond for municipal sewage treatment.

Anaerobic ponds have the potential to contribute to low carbon wastewater treatment, however are currently restricted by long hydraulic residence time (HRT) which leads to large land requirements. A two-stage anaerobic pond (SAP) design was trialled against a single-stage control (CAP) over four HRTs down to 0.5 days, to determine the lowest HRT at which the ponds could operate effectively. No statistical differences were observed in particulate removal between the ponds over all four HRTs, suggesting solids loading is not a critical factor in AP design. Significantly higher biogas production rates were observed in the SAP than the CAP at 1.5 d and 1.0 d HRT, and microbial community profiling suggests the two-stage design may be facilitating spatial separation of the anaerobic digestion process along reactor length. Hydrogenotrophic methanogensis dominated over aceticlastic, with acetate oxidisation a likely degradation pathway. Experimental tracer studies were compared to CFD simulations, with the SAP showing greater hydraulic efficiency, and differences more pronounced at shorter HRTs. Greater flow recirculation between baffles was observed in CFD velocity profiles, demonstrating baffles can dissipate preferential flow patterns and increase effective pond volume, especially at high flow rates. The study demonstrates the potential of APs to be operated at shorter HRTs in psychrophilic conditions, presenting an opportunity for use as pre-treatments (in place of septic tanks) and primary treatment for full wastewater flows. Two-stage designs should be investigated to separate the stages of the anaerobic digestion process by creating preferential conditions along the pond length.

Managing power dissipation in closed-loop reverse electrodialysis to maximise energy recovery during thermal-to-electric conversion.

Whilst the efficiency of reverse electrodialysis (RED) for thermal-to-electrical conversion has been theoretically demonstrated for low-grade waste heat, the specific configuration and salinity required to manage power generation has been less well described. This study demonstrates that operating RED by recycling feed solutions provides the most suitable configuration for energy recovery from a fixed solution volume, providing a minimum unitary cost for energy production. For a fixed membrane area, recycling feeds achieves energy efficiency seven times higher than single pass (conventional operation), and with an improved power density. However, ionic transport, water flux and concentration polarisation introduce complex temporal effects when concentrated brines are recirculated, that are not ordinarily encountered in single pass systems. Regeneration of the concentration gradient at around 80% energy dissipation was deemed most economically pragmatic, due to the increased resistance to mass transport beyond this threshold. However, this leads to significant exergy destruction that could be improved by interventions to better control ionic build up in the dilute feed. Further improvements to energy efficiency were fostered through optimising current density for each brine concentration independently. Whilst energy efficiency was greatest at lower brine concentrations, the work produced from a fixed volume of feed solution was greatest at higher saline concentrations. Since the thermal-to-electrical conversion proposed is governed by volumetric heat utilisation (distillation to reset the concentration gradient), higher brine concentrations are therefore recommended to improve total system efficiency. Importantly, this study provides new evidence for the configuration and boundary conditions required to realise RED as a practical solution for application to sources of low-grade waste heat in industry.

Ultrafiltration pretreatment enhances membrane distillation flux, resilience and permeate quality during water recovery from concentrated blackwater (urine/faeces).

In this study, the pretreatment of concentrated blackwater using ultrafiltration (UF) was shown to improve the permeability, selectivity and robustness of membrane distillation (MD) for application to wastewater treatment. Concentrated blackwater comprises urine and faeces, with minimal flushwater added. The faecal contribution increased the soluble organic fraction and introduced coarse and colloidal particles into the urine, which increased resistance to filtration during dead-end UF. Ultrafiltration removed the particulate and colloidal fractions (MW > 500 kDa) from the blackwater, which permitted similar permeability and robustness for MD to that observed with urine (29.9 vs 25.9 kg m-2 h-1), which comprises a lower colloidal organic concentration. Without UF pretreatment, a higher density organic layer formed on the MD surface (197 vs 70 gCOD m-2) which reduced mass transfer, and transformed the contact angle from hydrophobic to hydrophilic (144.9° to 49.8°), leading to pore wetting and a dissipation in product water quality due to breakthrough. In comparison, with UF pretreatment, MD delivered permeate water quality to standards satisfactory for discharge or reuse. This is particularly timely as the ISO standard for non-sewered sanitation has been adopted by several countries at a national level, and to date there are relatively few technologies to achieve the treatment standard. Membrane distillation provides a robust means for concentrated blackwater treatment, and since the energy required for separation is primarily heat, this advanced treatment can be delivered into areas with more fragile power networks.

Establishing the mechanisms underpinning solids breakthrough in UASB configured anaerobic membrane bioreactors to mitigate fouling.

In this study, the mechanisms for solids breakthrough in upflow anaerobic sludge blanket (UASB) configured anaerobic membrane bioreactors (AnMBRs) have been described to establish design parameters to limit membrane fouling. As the sludge blanket develops, two periods can be identified: (i) an initial progressive enhancement in solids separation provided through sludge blanket clarification, via depth filtration, which sustains downstream membrane permeability; and (ii) sludge blanket destabilisation, which imposed solids breakthrough resulting in a loss in membrane permeability. The onset of sludge blanket destabilisation was identified earlier in the flocculent AnMBR, which was ascribed to an increased gas production, caused by hydrolysis within the sludge blanket at extended solids residence time. Whilst hydrolysis also induced higher gas productivity within the granular AnMBR, solids breakthrough was not evidently observed during this period, and was instead only observed as the sludge blanket approached the UASB overflow. However, solids breakthrough was observed earlier for both reactors when treating wastewater with lower temperatures. This was explained through characterisation of the settling velocity of discrete particles from the sludge blanket of both MBRs; solids washout was evidenced to be induced by the increase in fluid viscosity with a reduction in temperature, which lowered terminal particle settling velocity. Nevertheless, particle settling velocity was comparable for particles from both sludge blankets. We therefore propose that the enhanced stability imparted by the granular AnMBR is due to the higher inertial force of the dense granular sludge. From this study, we suggest that similarly low levels of membrane fouling can be achieved within flocculent AnMBR by managing solids retention time to constrain sludge bed height and excess hydrolysis, together with adopting an upflow velocity based on particle buoyancy at the lowest expected operating temperature.

Non-isothermal drying kinetics of human feces.

The non-isothermal drying behavior and kinetics of human feces (HF) were investigated by means of thermogravimetric analysis to provide data for designing a drying unit operation. The effect of heating rate and blending with woody biomass were also evaluated on drying pattern and kinetics. At low heating rate (1 K/min), there is effective transport of moisture, but a higher heating rate would be necessary at low moisture levels to reduce drying time. Blending with wood biomass improves drying characteristics of HF. The results presented in this study are relevant for designing non-sewered sanitary systems with in-situ thermal treatment.

Hybrid membrane distillation reverse electrodialysis configuration for water and energy recovery from human urine: An opportunity for off-grid decentralised sanitation.

The integration of membrane distillation with reverse electrodialysis has been investigated as a sustainable sanitation solution to provide clean water and electrical power from urine and waste heat. Reverse electrodialysis was integrated to provide the partial remixing of the concentrate (urine) and diluate (permeate) produced from the membrane distillation of urine. Broadly comparable power densities to those of a model salt solution (sodium chloride) were determined during evaluation of the individual and combined contribution of the various monovalent and multivalent inorganic and organic salt constituents in urine. Power densities were improved through raising feed-side temperature and increasing concentration in the concentrate, without observation of limiting behaviour imposed by non-ideal salt and water transport. A further unique contribution of this application is the limited volume of salt concentrate available, which demanded brine recycling to maximise energy recovery analogous to a battery, operating in a 'state of charge'. During recycle, around 47% of the Gibbs free energy was recoverable with up to 80% of the energy extractable before the concentration difference between the two solutions was halfway towards equilibrium which implies that energy recovery can be optimised with limited effect on permeate quality. This study has provided the first successful demonstration of an integrated MD-RED system for energy recovery from a limited resource, and evidences that the recovered power is sufficient to operate a range of low current fluid pumping technologies that could help deliver off-grid sanitation and clean water recovery at single household scale.

Sustainable removal of N2O by mediated electrocatalytic reduction at ambient temperature electro-scrubbing using electrogenerated Ni(I) electron mediator.

Direct catalysis is generally proposed for nitrous oxide (N2O) abatement but catalysis is expensive, requires high temperatures, and suffers from media fouling, which limits its lifetime. In the present study, an ambient temperature electroscrubbing method was developed, coupling wet-scrubbing with an electrogenerated Ni(I) ([Ni(I)(CN)4]3-) mediator, to enable N2O reduction in a single process stage. The initial studies of 10 ppm N2O absorption into 9 M KOH and an electrolyzed 9 M KOH solution showed no removal. However, 95% N2O removal was identified through the addition of Ni(I) to an electrolyzed 9 M KOH. A change in the oxidation/reduction potential from -850 mV to -650 mV occurred following a decrease in Ni(I) concentration from 4.6 mM to 4.0 mM, which confirmed that N2O removal was mediated by an electrocatalytic reduction (MER) pathway. Online analysis identified the reaction product to be ammonia (NH3). Increasing the feed N2O concentration increased NH3 formation, which suggests that a decrease in electrolyzed solution reactivity induced by the increased N2O load constrained the side reaction with the carrier gas. Importantly, this study outlines a new regenerable method for N2O removal to commodity product NH3 at ambient temperature that fosters process intensification, overcomes the limitations generally observed with catalysis, and permits product transformation to NH3.

Non-isothermal thermogravimetric kinetic analysis of the thermochemical conversion of human faeces.

The "Reinvent the Toilet Challenge" set by the Bill & Melinda Gates Foundation aims to bring access to adequate sanitary systems to billions of people. In response to this challenge, on-site sanitation systems are proposed and being developed globally. These systems require in-situ thermal treatment, processes that are not well understood for human faeces (HF). Thermogravimetric analysis has been used to investigate the pyrolysis, gasification and combustion of HF. The results are compared to the thermal behaviour of simulant faeces (SF) and woody biomass (WB), along with the blends of HF and WB. Kinetic analysis was conducted using non-isothermal kinetics model-free methods, and the thermogravimetric data obtained for the combustion of HF, SS and WB. The results show that the devolatilisation of HF requires higher temperatures and rates are slower those of WB. Minimum temperatures of 475 K are required for fuel ignition. HF and SF showed similar thermal behaviour under pyrolysis, but not under combustion conditions. The activation energy for HF is 157.4 kJ/mol, relatively higher than SS and WB. Reaction order for HF is lower (n = 0.4) to WB (n = 0.6). In-situ treatment of HF in on-site sanitary systems can be designed for slow progressive burn.

Electrochemically generated bimetallic reductive mediator Cu1+[Ni2+(CN)4]1- for the degradation of CF4 to ethanol by electro-scrubbing.

Remediation of electronic gas CF4 using commercially available technologies results in another kind of greenhouse gas and corrosive side products. This investigation aimed to develop CF4 removal at room temperature with formation of useful product by attempting an electrogenerated Cu1+[Ni2+(CN)4]1- mediator. The initial electrolysis of the bimetallic complex at the anodized Ti cathode demonstrated Cu1+[Ni2+(CN)4]1- formation, which was confirmed by additional electron spin resonance results. The degradation of CF4 followed mediated electrochemical reduction by electrogenerated Cu1+[Ni2+(CN)4]1-. The removal efficiency of CF4 of 95% was achieved by this electroscrubbing process at room temperature. The spectral results of online and offline Fourier transform infrared analyzer, either in gas or in solution phase, demonstrated that the product formed during the removal of CF4 by electrogenerated Cu1+[Ni2+(CN)4]1- by electroscrubbing was ethanol (CH3CH2OH), with a small amount of trifluoroethane (CF3CH3) intermediate.

Impact of fouling, cleaning and faecal contamination on the separation of water from urine using thermally driven membrane separation.

In this study, membrane distillation is evaluated as a technology for non-sewered sanitation, using waste heat to enable separation of clean water from urine. Whilst membrane fouling was observed for urine, wetting was not evident and product water quality met the proposed discharge standard, despite concentration of the feed. Fouling was reversible using physical cleaning, which is similar to previous membrane studies operating without pressure as the driving force. High COD reduction was achieved following faecal contamination, but mass transfer was impeded and wetting occurred which compromised permeate quality, suggesting upstream intervention is demanded to limit the extent of faecal contamination. (100 words).

Tube-side mass transfer for hollow fibre membrane contactors operated in the low Graetz range.

Transformation of the tube-side mass transfer coefficient derived in hollow fibre membrane contactors (HFMC) of different characteristic length scales (equivalent diameter and fibre length) has been studied when operated in the low Graetz range (Gz<10). Within the low Gz range, mass transfer is generally described by the Graetz problem (Sh=3.67) which assumes that the concentration profile comprises a constant shape over the fibre radius. In this study, it is experimentally evidenced that this assumption over predicts mass transfer within the low Graetz range. Furthermore, within the low Gz range (below 2), a proportional relationship between the experimentally determined mass transfer coefficient (Kov ) and the Graetz number has been identified. For Gz numbers below 2, the experimental Sh number approached unity, which suggests that mass transfer is strongly dependent upon diffusion. However, within this diffusion controlled region of mass transfer, tube-side fluid velocity remained important. For Gz numbers above 2, Sh could be satisfactorily described by extension to the Lévêque solution, which can be ascribed to the constrained growth of the concentration boundary layer adjacent to the fibre wall. Importantly this study demonstrates that whilst mass transfer in the low Graetz range does not explicitly conform to either the Graetz problem or classical Lévêque solution, it is possible to transform the experimentally derived overall mass transfer coefficient (Kov ) between characteristic length scales (dh and L). T h is was corroborated by comparison of the empirical relationship determined in this study (Sh=0.36Gz) with previously published studies operated in the low Gz range. This analysis provides important insight for process design when slow tube-side flows, or low Schmidt numbers (coincident with gases) constrain operation of hollow fibre membrane contactors to the low Gz range.

Energy recovery from human faeces via gasification: A thermodynamic equilibrium modelling approach.

Non-sewered sanitary systems (NSS) are emerging as one of the solutions to poor sanitation because of the limitations of the conventional flush toilet. These new sanitary systems are expected to safely treat faecal waste and operate without external connections to a sewer, water supply or energy source. The Nano Membrane Toilet (NMT) is a unique domestic-scale sanitary solution currently being developed to treat human waste on-site. This toilet will employ a small-scale gasifier to convert human faeces into products of high energy value. This study investigated the suitability of human faeces as a feedstock for gasification. It quantified the recoverable exergy potential from human faeces and explored the optimal routes for thermal conversion, using a thermodynamic equilibrium model. Fresh human faeces were found to have approximately 70-82 wt.% moisture and 3-6 wt.% ash. Product gas resulting from a typical dry human faeces (0 wt.% moisture) had LHV and exergy values of 17.2 MJ/kg and 24 MJ/kg respectively at optimum equivalence ratio of 0.31, values that are comparable to wood biomass. For suitable conversion of moist faecal samples, near combustion operating conditions are required, if an external energy source is not supplied. This is however at 5% loss in the exergy value of the gas, provided both thermal heat and energy of the gas are recovered. This study shows that the maximum recoverable exergy potential from an average adult moist human faeces can be up to 15 MJ/kg, when the gasifier is operated at optimum equivalence ratio of 0.57, excluding heat losses, distribution or other losses that result from operational activities.

The cost of a small membrane bioreactor.

The individual cost contributions to the mechanical components of a small membrane bioreactor (MBR) (100-2,500 m3/d flow capacity) are itemised and collated to generate overall capital and operating costs (CAPEX and OPEX) as a function of size. The outcomes are compared to those from previously published detailed cost studies provided for both very small containerised plants (<40 m3/day capacity) and larger municipal plants (2,200-19,000 m3/d). Cost curves, as a function of flow capacity, determined for OPEX, CAPEX and net present value (NPV) based on the heuristic data used indicate a logarithmic function for OPEX and a power-based one for the CAPEX. OPEX correlations were in good quantitative agreement with those reported in the literature. Disparities in the calculated CAPEX trend compared with reported data were attributed to differences in assumptions concerning cost contributions. More reasonable agreement was obtained with the reported membrane separation component CAPEX data from published studies. The heuristic approach taken appears appropriate for small-scale MBRs with minimal costs associated with installation. An overall relationship of net present value=(a tb)Q(-c lnt+d) was determined for the net present value where a=1.265, b=0.44, c=0.00385 and d=0.868 according to the dataset employed for the analysis.

Development of on-line FTIR spectroscopy for siloxane detection in biogas to enhance carbon contactor management.

Activated carbon filters are used to limit engine damage by siloxanes when biogas is utilised to provide electricity. However, carbon filter siloxane removal performance is poorly understood as until recently, it had not been possible to measure siloxanes on-line. In this study, on-line Fourier Transform Infrared (FTIR) spectroscopy was developed to measure siloxane concentration in real biogas both upstream (86.1-157.5mg m(-3)) and downstream (2.2-4.3mg m(-3)) of activated carbon filters. The FTIR provided reasonable precision upstream of the carbon vessel with a root mean square error of 10% using partial least squares analysis. However, positive interference from volatile organic carbons was observed in downstream gas measurements limiting precision at the outlet to an RMSE of 1.5mg m(-3) (47.8%). Importantly, a limit of detection of 3.2mg m(-3) was identified which is below the recommended siloxane limit and evidences the applicability of on-line FTIR for this application.

Characterization of full-scale carbon contactors for siloxane removal from biogas using online Fourier transform infrared spectroscopy.

In this study, online Fourier transform infrared (FTIR) spectroscopy has been used to generate the first comprehensive characterization of full-scale carbon contactors for siloxane removal from biogas. Using FTIR, two clear operational regions within the exhaustion cycle were evidenced: an initial period of pseudo-steady state where the outlet siloxane concentration was consistently below the proposed siloxane limits; and a second period characterized by a progressive rise in outlet siloxane concentration during and after breakthrough. Due to the sharp breakthrough front identified, existing detection methods (which comprise field sampling coupled with laboratory-based chromatographic determination) are insufficiently responsive to define breakthrough, thus carbon contactors currently remain in service while providing limited protection to the combined heat and power engine. Integration of the exhaustion cycle to breakthrough identified average specific media capacities of 8.5-21.5 gsiloxane kg(-1)GAC, which are lower than that has been reported for vapour phase granular activated carbon (GAC). Further speciation of the biogas phase identified co-separation of organic compounds (alkanes and aromatics), which will inevitably reduce siloxane capacity. However, comparison of the five full-scale contactors identified that greater media capacity was accessible through operating contactors at velocities sufficient to diminish axial dispersion effects. In addition to enabling significant insight into gas phase GAC contactors, the use of FTIR for online control of GAC for siloxane removal is also presented.

Reflux composition influences the level of NF-κB activation and upstream kinase preference in oesophageal adenocarcinoma cells.

Oesophageal adenocarcinoma (OA) incidence is rising and prognosis is poor. Understanding the molecular basis of this malignancy is key to finding new prevention and treatment strategies. Gastroesophageal reflux disease is the primary cause of OA, usually managed with acid suppression therapy. However, this often does little to control carcinogenic bile acid reflux. The transcription factor nuclear factor kappa B (NF-κB) plays a key role in the pathogenesis of OA and its activity is associated with a poor response to chemotherapy, making it an attractive therapeutic target. We sought to decipher the role of different bile acids in NF-κB activation in oesophageal cell lines using short, physiologically relevant exposure times. The effect of an acidic or neutral extracellular pH was investigated concurrently, to mimic in vivo conditions associated with or without acid suppression. We found that some bile acids activated NF-κB to a greater extent when combined with acid, whereas others did so in its absence, at neutral pH. The precise composition of an individual's reflux, coupled with whether they are taking acid suppressants may therefore dictate the extent of NF-κB activation in the oesophagus, and hence the likelihood of histological progression and chemotherapy success. Regardless of pH, the kinase inhibitor of κB kinase was pivotal in mediating reflux induced NF-κB activation. Its importance was confirmed further as its increased activation was associated with histological progression in patient samples. We identified further kinases important in acid or bile induced NF-κB signalling in oesophageal cells, which may provide suitable targets for therapeutic intervention.

Inducible nitric oxide synthase (iNOS) and nitric oxide (NO) are important mediators of reflux-induced cell signalling in esophageal cells.

Nitric oxide (NO) produced by inducible nitric oxide synthase (iNOS) has been implicated in both DNA damage induction and aberrant cell signalling in various tissue and cell backgrounds. We investigated here the role of iNOS and NO in DNA damage induction and nuclear factor-kappa B (NF-κB) signalling in esophageal cells in vitro. As esophageal adenocarcinoma develops in a background of Barrett's esophagus secondary to reflux disease, it is possible that inflammatory mediators like NO may be important in esophageal cancer development. We show that reflux components like stomach acid and bile acids [deoxycholic acid (DCA)] can induce iNOS gene and protein expression and produce NO generation in esophageal cells, using real-time PCR, western blotting and NO sensitive fluorescent probes, respectively. This up-regulation of iNOS expression was not dependent on NF-κB activity. DCA-induced DNA damage was independent of NF-κB and only partially dependent on iNOS and NO, as measured by the micronucleus assay. These same reflux constituents also activated the oncogenic transcription factor NF-κB, as measured by transcription factor enzyme-linked immunosorbent assay and gene expression studies with NF-κB linked genes (e.g. interleukin-8). Importantly, we show here for the first time that basal levels of NF-κB activity (and possibly acid and DCA-induced NF-κB) are dependent on iNOS/NO and this may lead to a positive feedback loop whereby induced iNOS is upstream of NF-κB, hence prolonging and potentially amplifying this signalling, presumably through NO activation of NF-κB. Furthermore, we confirm increased protein levels of iNOS in esophageal adenocarcinoma and, therefore, in neoplastic development in the esophagus.