Modulating the Cellular Uptake of Fluorescently Tagged Substrates of Prostate-Specific Antigen before and after Enzymatic Activation.

A series of peptides based on the prostate-specific antigen (PSA)-specific sequence histidine-serine-serine-lysine-leucine-glutamine were functionalized with an anthraquinone fluorophore at the C-terminal residue side chain using the copper(I)-catalyzed azide-alkyne cycloaddition reaction. The effect of incorporating a negatively charged N-terminal tetra-glutamic acid group into the substrate and the effect of masking the negatively charged C-terminal carboxylic acid functionality of the substrate were investigated using confocal fluorescence microscopy in two cell lines, DLD-1 and LnCaP. The addition of a tetra-glutamic acid group to the N-terminus of the intact sequence was shown to reduce cellular uptake of the intact substrate prior to activation by PSA. In contrast, masking the C-terminal carboxylic acid group of the substrate as a methyl ester was shown to improve cellular uptake of the peptide fragment after activation by PSA. The synthesized C-terminal methyl ester substrates with the anthraquinone attached to the side chain were confirmed to be cleaved by PSA in LC-MS analysis, and the cytotoxicity of the substrates was shown to increase in the presence of PSA, consistent with cleavage and uptake of the C-terminal fragment. The results indicate that C- and N-terminal functionalization of peptide substrates targeting PSA can be used to modulate the cellular uptake of peptides before and after enzymatic activation, which may thus be an important consideration in the design of tumor-activated prodrugs.

Biodistribution and Clearance of Stable Superparamagnetic Maghemite Iron Oxide Nanoparticles in Mice Following Intraperitoneal Administration.

Nanomedicine is an emerging field with great potential in disease theranostics. We generated sterically stabilized superparamagnetic iron oxide nanoparticles (s-SPIONs) with average core diameters of 10 and 25 nm and determined the in vivo biodistribution and clearance profiles. Healthy nude mice underwent an intraperitoneal injection of these s-SPIONs at a dose of 90 mg Fe/kg body weight. Tissue iron biodistribution was monitored by atomic absorption spectroscopy and Prussian blue staining. Histopathological examination was performed to assess tissue toxicity. The 10 nm s-SPIONs resulted in higher tissue-iron levels, whereas the 25 nm s-SPIONs peaked earlier and cleared faster. Increased iron levels were detected in all organs and body fluids tested except for the brain, with notable increases in the liver, spleen, and the omentum. The tissue-iron returned to control or near control levels within 7 days post-injection, except in the omentum, which had the largest and most variable accumulation of s-SPIONs. No obvious tissue changes were noted although an influx of macrophages was observed in several tissues suggesting their involvement in s-SPION sequestration and clearance. These results demonstrate that the s-SPIONs do not degrade or aggregate in vivo and intraperitoneal administration is well tolerated, with a broad and transient biodistribution. In an ovarian tumor model, s-SPIONs were shown to accumulate in the tumors, highlighting their potential use as a chemotherapy delivery agent.

Tunable and noncytotoxic PET/SPECT-MRI multimodality imaging probes using colloidally stable ligand-free superparamagnetic iron oxide nanoparticles.

Physiologically stable multimodality imaging probes for positron emission tomography/single-photon emission computed tomography (PET/SPECT)-magnetic resonance imaging (MRI) were synthesized using the superparamagnetic maghemite iron oxide (γ-Fe2O3) nanoparticles (SPIONs). The SPIONs were sterically stabilized with a finely tuned mixture of diblock copolymers with either methoxypolyethylene glycol (MPEG) or primary amine NH2 end groups. The radioisotope for PET or SPECT imaging was incorporated with the SPIONs at high temperature. 57Co2+ ions with a long half-life of 270.9 days were used as a model for the radiotracer to study the kinetics of radiolabeling, characterization, and the stability of the radiolabeled SPIONs. Radioactive 67Ga3+ and Cu2+-labeled SPIONs were also produced successfully using the optimized conditions from the 57Co2+-labeling process. No free radioisotopes were detected in the aqueous phase for the radiolabeled SPIONs 1 week after dispersion in phosphate-buffered saline (PBS). All labeled SPIONs were not only well dispersed and stable under physiological conditions but also noncytotoxic in vitro. The ability to design and produce physiologically stable radiolabeled magnetic nanoparticles with a finely controlled number of functionalizable end groups on the SPIONs enables the generation of a desirable and biologically compatible multimodality PET/SPECT-MRI agent on a single T2 contrast MRI probe.

Open burning and open detonation PM10 mass emission factor measurements with optical remote sensing.

Emission factors (EFs) of particulate matter with aerodynamic diameter <10 microm (PM10) from the open burning/open detonation (OB/OD) of energetic materials were measured using a hybrid-optical remote sensing (hybrid-ORS) method. This method is based on the measurement of range-resolved PM backscattering values with a micropulse light detection and ranging (LIDAR; MPL) device. Field measurements were completed during March 2010 at Tooele Army Depot, Utah, which is an arid continental site. PM10 EFs were quantified for OB of M1 propellant and OD of 2,4,6-trinitrotoluene (TNT). EFs from this study are compared with previous OB/OD measurements reported in the literature that have been determined with point measurements either in enclosed or ambient environments, and with concurrent airborne point measurements. PM10 mass EFs, determined with the hybrid-ORS method, were 7.8 x 10(-3) kg PM10/kg M1 from OB of M1 propellant, and 0.20 kg PM10/kg TNT from OD of TNT. Compared with previous results reported in the literature, the hybrid-ORS method EFs were 13% larger for OB and 174% larger for OD. Compared with the concurrent airborne measurements, EF values from the hybrid-ORS method were 37% larger for OB and 54% larger for OD. For TNT, no statistically significant differences were observed for the EFs measured during the detonation of 22.7 and 45.4 kg of TNT, supporting that the total amount of detonated mass in this mass range does not have an effect on the EFs for OD of TNT.

Simultaneous removal of perchlorate and energetic compounds in munitions wastewater by zero-valent iron and perchlorate-respiring bacteria.

Ammonium perchlorate is one of the main constituents in Army's insensitive melt-pour explosive, PAX-21 in addition to RDX and 2,4-dinitroanisole (DNAN). The objective of this study is to develop an innovative treatment process to remove both perchlorate and energetic compounds simultaneously from PAX-21 production wastewater. It was hypothesized that the pretreatment of PAX-21 wastewater with zero-valent iron (ZVI) would convert energetic compounds to products that are more amenable for biological oxidation and that these products serve as electron donors for perchlorate-reducing bacteria. Results of batch ZVI reduction experiments showed that DNAN was completely reduced to 2,4-diaminoanisole and RDX was completely reduced to formaldehyde. Anaerobic batch biodegradation experiments showed that perchlorate (30 mg L(-1)) in ZVI-treated PAX-21 wastewater was decreased to an undetectable level after 5 days. Batch biodegradation experiments also confirmed that formaldehyde in ZVI-treated wastewater was the primary electron donor for perchlorate-respiring bacteria. The integrated iron-anaerobic bioreactor system was effective in completely removing energetic compounds and perchlorate from the PAX-21 wastewater without adding an exogenous electron donor. This study demonstrated that ZVI pretreatment not only removed energetic compounds, but also transformed energetic compounds to products that can serve as the source of electrons for perchlorate-respiring bacteria.

The role of waixenicin A as transient receptor potential melastatin 7 blocker.

Transient receptor potential melastatin 7 (TRPM7) plays a role in a number of physiological and pharmacological functions in variety of cells. The aim of this study was to clarify the role for TRPM7 channels and the effect of waixenicin A on the pacemaking activity of interstitial cells of Cajal (ICCs) and on the cell viability of the human gastric and breast adenocarcinoma cell lines, AGS and MCF-7, respectively. Waixenicin A decreased the amplitude of pacemaker potentials in cultured ICC clusters and inhibited TRPM7 currents, but had no effect on Ca(2+) -activated Cl(-) conductance (ANO1). Furthermore, waixenicin A was found to inhibit the growth and survival of AGS and MCF-7 cells. These findings indicate that TRPM7 channel modulates intestinal motility and regulates the pathophysiology of human gastric and breast adenocarcinoma cells. These findings suggest that TRPM7 channel be considered a potential target for the treatment of gut motor disorders and gastric and breast cancer.

Dual targeting of hypoxic and acidic tumor environments with a cobalt(III) chaperone complex.

The rational design of prodrugs for selective accumulation and activation in tumor microenvironments is one of the most promising strategies for minimizing the toxicity of anticancer drugs. Manipulation of the charge of the prodrug represents a potential mechanism to selectively deliver the prodrug to the acidic tumor microenvironment. Here we present delivery of a fluorescent coumarin using a cobalt(III) chaperone to target hypoxic regions, and charged ligands for pH selectivity. Protonation or deprotonation of the complexes over a physiologically relevant pH range resulted in pH dependent accumulation of the fluorophore in colon cancer cells. Furthermore, in a spheroid solid tumor model, the anionic complexes exhibited preferential release of the fluorophore in the acidic/hypoxic region. By fine-tuning the physicochemical properties of the cobalt-chaperone moiety, we have demonstrated selective drug release in the acidic and hypoxic tumor microenvironment.

Cobalt complexes with tripodal ligands: implications for the design of drug chaperones.

Extensive research is currently being conducted into metal complexes that can selectively deliver cytotoxins to hypoxic regions in tumours. The development of pharmacologically suitable agents requires an understanding of appropriate ligand-metal systems for chaperoning cytotoxins. In this study, cobalt complexes with tripodal tren (tris-(2-aminoethyl)amine) and tpa (tris-(2-pyridylmethyl)amine) ligands were prepared with ancillary hydroxamic acid, ß-diketone and catechol ligands and several parameters, including: pK(a), reduction potential and cytotoxicity were investigated. Fluorescence studies demonstrated that only tpa complexes with ß-diketones showed any reduction by ascorbate in situ and similarly, cellular cytotoxicity results demonstrated that ligation to cobalt masked the cytotoxicity of the ancillary groups in all complexes except the tpa diketone derivative [Co(naac)tpa](ClO(4))(2) (naac = 1-methyl-3-(2-naphthyl)propane-1,3-dione). Additionally, it was shown that the hydroxamic acid complexes could be isolated in both the hydroxamate and hydroximate form and the pK(a) values (5.3-8.5) reveal that the reversible protonation/deprotonation of the complexes occurs at physiologically relevant pHs. These results have clear implications for the future design of prodrugs using cobalt moieties as chaperones, providing a basis for the design of cobalt complexes that are both more readily reduced and more readily taken up by cells in hypoxic and acidic environments.

Detoxification of PAX-21 ammunitions wastewater by zero-valent iron for microbial reduction of perchlorate.

US Army and the Department of Defense (DoD) facilities generate perchlorate (ClO(4)(-)) from munitions manufacturing and demilitarization processes. Ammonium perchlorate is one of the main constituents in Army's new main charge melt-pour energetic, PAX-21. In addition to ammonium perchlorate, hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and 2,4-dinitroanisole (DNAN) are the major constituents of PAX-21. In order to evaluate microbial perchlorate reduction as a practical option for the treatment of perchlorate in PAX-21 wastewater, we conducted biodegradation experiments using glucose as the primary sources of electrons and carbon. Batch experiments showed that negligible perchlorate was removed in microbial reactors containing PAX-21 wastewater while control bottles containing seed bacteria and glucose rapidly and completely removed perchlorate. These results suggested that the constituents in PAX-21 wastewater may be toxic to perchlorate reducing bacteria. A series of batch toxicity test was conducted to identify the toxic constituents in PAX-21 and DNAN was identified as the primary toxicant responsible for inhibiting the activity of perchlorate reducing bacteria. It was hypothesized that pretreatment of PAX-21 by zero-valent iron granules will transform toxic constituents in PAX-21 wastewater to non-toxic products. We observed complete reduction of DNAN to 2,4-diaminoanisole (DAAN) and RDX to formaldehyde in abiotic iron reduction study. After a 3-day acclimation period, perchlorate in iron-treated PAX-21 wastewater was rapidly decreased to an undetectable level in 2 days. This result demonstrated that iron treatment not only removed energetic compounds but also eliminated the toxic constituents that inhibited the subsequent microbial process.

Optical remote sensing to quantify fugitive particulate mass emissions from stationary short-term and mobile continuous sources: part I. Method and examples.

The emissions of particulate matter (PM) from anthropogenic sources raise public concern. A new method is described here that was developed to complete in situ rapid response measurements of PM mass emissions from fugitive dust sources by use of optical remote sensing (ORS) and an anemometer. The ORS system consists of one ground-based micropulse light detection and ranging (MPL) device that was mounted on a positioner, two open path-Fourier transform infrared (OP-FTIR) spectrometers, and two open path-laser transmissometers (OP-LT). An algorithm was formulated to compute PM light extinction profiles along each of the plume's cross sections that were determined with the MPL. Size-specific PM mass emission factors were then calculated by integrating the light extinction profiles with particle mass extinction efficiencies (determined with the OP-FTIRs/OP-LTs) and the wind's speed and direction. This method also quantifies the spatial and temporal variability of the plume's PM mass concentrations across each of the plume's cross sections. Example results from three field studies are also described to demonstrate how this new method is used to determine mass emission factors as well as characterize the dust plumes' horizontal and vertical dimensions and temporal variability of the PM's mass concentration.

Optical remote sensing to quantify fugitive particulate mass emissions from stationary short-term and mobile continuous sources: part II. Field applications.

Quantification of emissions of fugitive particulate matter (PM) into the atmosphere from military training operations is of interest by the United States Department of Defense. A new range-resolved optical remote sensing (ORS) method was developed to quantify fugitive PM emissions from puff sources (i.e., artillery back blasts), ground-level mobile sources (i.e., movement of tracked vehicles), and elevated mobile sources (i.e., airborne helicopters) in desert areas that are prone to generating fugitive dust plumes. Real-time, in situ mass concentration profiles for PM mass with particle diameters <10 µm (PM(10)) and <2.5 µm (PM(2.5)) were obtained across the dust plumes that were generated by these activities with this new method. Back blasts caused during artillery firing were characterized as a stationary short-term puff source whose plumes typically dispersed to <10 m above the ground with durations of 10-30 s. Fugitive PM emissions caused by artillery back blasts were related to the zone charge and ranged from 51 to 463 g PM/firing for PM(10) and 9 to 176 g PM/firing for PM(2.5). Movement of tracked vehicles and flying helicopters was characterized as mobile continuous sources whose plumes typically dispersed 30-50 m above the ground with durations of 100-200 s. Fugitive PM emissions caused by moving tracked vehicles ranged from 8.3 to 72.5 kg PM/km for PM(10) and 1.1 to 17.2 kg PM/km for PM(2.5), and there was no obvious correlation between PM emission and vehicle speed. The emission factor for the helicopter flying at 3 m above the ground ranged from 14.5 to 114.1 kg PM/km for PM(10) and 5.0 to 39.5 kg PM/km for PM(2.5), depending on the velocity of the helicopter and type of soil it flies over. Fugitive PM emissions by an airborne helicopter were correlated with helicopter speed for a particular soil type. The results from this range-resolved ORS method were also compared with the data obtained with another path-integrated ORS method and a Flux Tower method.

Evaluation of digital optical method to determine plume opacity during nighttime.

United States Environmental Protection Agency (USEPA) set opacity standards for visual emissions from industrial sources to protect ambient air quality. USEPA developed Method 9, which is a reference method to describe how plume opacity can be quantified by human observers during daytime conditions. However, it would be beneficial to determine plume opacity with digital still cameras (DSCs) to provide graphical records of the plume and its environment during visual emission evaluation and to be able to determine plume opacity with DSCs during nighttime conditions. Digital optical method (DOM) was developed to quantify plume opacity from photographs that were provided by a DSC during daytime. Past daytime field campaigns have demonstrated that DOM provided opacity readings that met Method 9 certification requirements. In this paper, the principles and methodology of DOM to quantify plume opacity during nighttime are described. Also, results are described from a nighttime field campaign that occurred at Springfield, IL. Opacity readings provided by DOM were compared with the opacity values obtained with the reference in-stack transmissometer of the smoke generator. The average opacity errors were 2.3-3.5% for contrast model of DOM for all levels of plume opacity. The average opacity errors were 2.0-7.6% for the transmission model of DOM for plumes with opacity 0-50%. These results are encouraging and indicate that DOM has the potential to quantify plume opacity during nighttime.

Effect of gas empty bed contact time on performances of various types of rotating drum biofilters for removal of VOCs.

The effects of gas empty bed contact time (EBCT), biofilter configuration, and types of volatile organic compounds (VOCs) were evaluated to assess the performance of rotating drum biofilters (RDBs), especially at low EBCT values. Three types of pilot-scale RDBs, a single-layer RDB, a multi-layer RDB, and a hybrid RDB, were examined at various gas EBCTs but at a constant VOC loading rate. Diethyl ether, toluene, and hexane were used separately as model VOC. When EBCT increased from 5.0 to 60s at a constant VOC loading rate of 2.0kgCOD/(m(3)day), ether removal efficiency increased from 73.1% to 97.6%, from 81.6% to 99.9%, and from 84.0% to 99.9% for the single-layer RDB, the multi-layer RDB, and the hybrid RDB, respectively, and toluene removal efficiency increased from 76.4% to 99.9% and from 84.8% to 99.9% for the multi-layer RDB and the hybrid RDB, respectively. When hexane was used as the model VOC at a constant loading rate of 0.25kgCOD/(m(3)day), hexane removal efficiency increased from 31.1% to 57.0% and from 29.5% to 50.0% for the multi-layer RDB and hybrid RDB, respectively. The single-layer, multi-layer, and hybrid RDBs exhibited, respectively, the lowest, middle, and highest removal efficiencies, when operated under similar operational loading conditions. Hexane exhibited the lowest removal efficiency, while diethyl ether displayed the highest removal efficiency. The data collected at the various EBCT values correlated reasonably well with a saturation model. The sensitivity of removal efficiency to EBCT varied significantly with EBCT values, VOC properties, and biofilter configurations. Process selection and design for RDB processes should consider these factors.

Effect of carrier size on the performance of a three-phase circulating-bed biofilm reactor for removing toluene in gas stream.

A series of steady-state and short-term experiments on a three-phase circulating-bed biofilm reactor (CBBR) for removing toluene from gas streams were conducted to investigate the effect of macroporous-carrier size (1-mm cubes versus 4-mm cubes, which have the same total surface area) on CBBR performance. Experimental conditions were identical, except for the carrier size. The CBBR with 1-mm carriers (the 1-mm CBBR) overcame the performance limitation observed with the CBBR with 4-mm carriers (the 4-mm CBBR): oxygen depletion inside the biofilm. The 1-mm CBBR consistently had the superior removal efficiencies of toluene and COD, higher than 93% for all, and the advantage was greatest for the highest toluene loading, 0.12 M/m2-day. The 1-mm carriers achieved superior performance by minimizing the negative effects of oxygen depletion, because they had 4.7 to 6.8 times thinner biofilm depths. The 1-mm carriers continued to provide protection from excess biomass detachment and inhibition from toluene. Finally, the 1-mm CBBR achieved volumetric removal capacities up to 300 times greater than demonstrated by other biofilters treating toluene and related volatile hydrocarbons.

Concomitant adsorption and desorption of organic vapor in dry and humid air streams using microwave and direct electrothermal swing adsorption.

Industrial gas streams can contain highly variable organic vapor concentrations that need to be processed before they are emitted to the atmosphere. Fluctuations in organic vapor concentrations make it more difficult to operate a biofilter when compared to a constant vapor concentration. Hence, there is a need to stabilize the concentration of rapidly fluctuating gas streams for optimum operation of biofilters. This paper describes new concomitant adsorption desorption (CAD) systems used with variable organic vapor concentration gas streams to provide the same gas stream, but at a user-selected constant vapor concentration that can then be more readily processed by a secondary air pollution control device such as a biofilter. The systems adsorb organic vapor from gas streams and simultaneously heat the adsorbent using microwave or direct electrothermal energy to desorb the organic vapor at a user-selected set-point concentration. Both systems depicted a high degree of concentration stabilization with a mean relative deviation between set-point and stabilized concentration of 0.3-0.4%. The direct electrothermal CAD system was also evaluated to treat a humid gas stream (relative humidity = 85%) that contained a variable organic vapor concentration. The high humidity did not interfere with CAD operation as water vapor did not adsorb but penetrated through the adsorbent These results are important because they demonstrate the ability of CAD to effectively dampen concentration fluctuation in gas streams.

Steady-state and dynamic desorption of organic vapor from activated carbon with electrothermal swing adsorption.

A new method to achieve steady-state and dynamic-tracking desorption of organic compounds from activated carbon was developed and tested with a bench-scale system. Activated carbon fiber cloth (ACFC) was used to adsorb methyl ethyl ketone (MEK) from air streams. Direct electrothermal heating was then used to desorb the vapor to generate select vapor concentrations at 500 ppmv and 5000 ppmv in air. Dynamic-tracking desorption was also achieved with carefully controlled yet variable vapor concentrations between 250 ppmv and 5000 ppmv, while also allowing the flow rate of the carrier gas to change by 100%. These results were also compared to conditions when recovering MEK as a liquid, and using microwaves as the source of energy to regenerate the adsorbent to provide MEK as a vapor or a liquid.

Field evaluation of digital optical method to quantify the visual opacity of plumes.

Visual Determination of the Opacity of Emissions from Stationary Sources (Method 9) is a reference method established by U.S. Environmental Protection Agency (EPA) to quantify plume opacity. However, Method 9 relies on observations from humans, which introduces subjectivity. In addition, it is expensive to teach and certify personnel to evaluate plume opacity on a semiannual basis. In this study, field tests were completed during a "smoke school" and a 4-month monitoring program of plumes emitted from stationary sources with a Method 9 qualified observer to evaluate the use of digital photography and two computer algorithms as an alternative to Method 9. This Digital Optical Method (DOM) improves objectivity, costs less to implement than Method 9, and provides archival photographic records of the plumes. Results from "smoke school" tests indicate that DOM passed six of eight tests when the sun was located in the 140 degrees sector behind one of the three cameras, with the individual opacity errors of 15% or less and average opacity errors of 7.5% or less. DOM also passed seven of the eight tests when the sun was located in the 216 degrees sector behind another camera. However, DOM passed only one of the eight tests when the sun was located in the 116 degrees sector in front of the third camera. Certification to read plume opacity by a "smoke reader" for 6 months requires that the "smoke reader" pass one of the smoke school tests during smoke school. The average opacity errors and percentage of observations with individual opacity errors above 15% for the results obtained with DOM were lower than those obtained by the smoke school trainees with the sun was located behind the camera, whereas they were higher than the smoke school trainee results with the sun located in front of the camera. In addition, the difference between plume opacity values obtained by DOM and a Method 9 qualified observer, as measured in the field for two industrial sources, were 2.2%. These encouraging results demonstrate that DOM is able to meet Method 9 requirements under a wide variety of field conditions and, therefore, has potential to be used as an alternative to Method 9.

Rapid response concentration-controlled desorption of activated carbon to dampen concentration fluctuations.

Fluctuations in concentration of organic vapors in gas streams that are treated by devices such as biofilters or oxidizers make it challenging to remove the vapors from the gas streams in an efficient and economic manner. Combining adsorption with concentration-controlled desorption provides an active buffer between the source of vapors and the control device for better control of concentration and flow rate of the gas stream that is treated by the secondary control device, hence further enhancing the performance or reducing the size of the devices. Activated carbon fiber cloth is used with microwave swing adsorption to remove methyl ethyl ketone (MEK) from air streams and then provide a readily controllable feed stream of that vapor in air at a specified concentration and gas flow rate with steady-state tracking desorption. MEK was captured with >99.8% efficiency during the adsorption cycle. The MEK concentration during the regeneration cycle was readily controlled at concentration set-points between 170 and 5000 ppmv, within relative standard deviations of 1.8 and 4.9%, respectively, and at 20% of the gas flow rate that was treated during the adsorption cycle. Such capability of the system allows the secondary control device to be optimized for select constant concentrations and low gas flow rates that is not possible without such pretreatment.

Quantification of plume opacity by digital photography.

The United States Environmental Protection Agency (USEPA) developed Method 9 to describe how plume opacity can be quantified by humans. However, use of observations by humans introduces subjectivity, and is expensive due to semiannual certification requirements of the observers. The Digital Opacity Method (DOM) was developed to quantify plume opacity at lower cost, with improved objectivity, and to provide a digital record. Photographs of plumes were taken with a calibrated digital camera under specified conditions. Pixel values from those photographs were then interpreted to quantify the plume's opacity using a contrast model and a transmission model. The contrast model determines plume opacity based on pixel values that are related to the change in contrast between two backgrounds that are located behind and next to the plume. The transmission model determines the plume's opacity based on pixel values that are related to radiances from the plume and its background. DOM was field tested with a smoke generator. The individual and average opacity errors of DOM were within the USEPA Method 9 acceptable error limits for both field campaigns. Such results are encouraging and support the use of DOM as an alternative to Method 9.

Microbial reduction of perchlorate with zero-valent iron.

Microbial reduction of perchlorate in the presence of zero-valent iron was examined in both batch and column reactors to assess the potential of iron as the electron donor for biological perchlorate reduction process. Iron-supported mixed cultures completely removed 65 mg/L of perchlorate in batch reactors in 8 days. The removal rate was similar to that observed with hydrogen gas (5%) and acetate (173 mg/L) as electron donors. Repeated spiking of perchlorate to batch reactors containing iron granules and microorganisms showed that complete perchlorate reduction by the iron-supported culture was sustained over a long period. Complete removal of perchlorate by iron-supported anaerobic culture was also achieved in a bench-scale iron column with a hydraulic residence time of 2 days. This study demonstrated the potential applicability of zero-valent iron as a source of electrons for biological perchlorate reduction. Use of zero-valent iron may eliminate the need to continually supply electron donors such as organic substrates or explosive hydrogen gas. In addition, iron is inexpensive, safe to handle, and does not leave organic residuals in the treated water.