Infection Rate in Type I Open Distal Radius Fractures Surgically Treated >24 Hours Post-Injury: A Comparison Study.

PURPOSE: To determine how time to surgical debridement and fixation affects infection and complication rate in type I open distal radius fractures by comparing patients treated within 24 hours with those treated >24 hours post-injury. METHODS: A retrospective review identified 62 patients who sustained a type I open distal radius fracture that was treated surgically. Patients were stratified into groups based on time to surgical intervention. An additional analysis was performed on patients with an isolated type I open distal radius fracture treated as an inpatient or outpatient. The primary outcome measure was infection rate. Secondary outcome measures were complications, reoperations, and readmissions. RESULTS: Thirty-eight patients underwent surgery ≤24 hours post-injury at an average of 14 hours. Twenty-four patients underwent surgery >24 hours post-injury at an average of 72 hours. There were a total of 9 complications in 8 patients (14.5%). The overall infection rate was 1.6%, with only 1 deep infection occurring in the group treated ≤24 hours post-injury. There were 7 reoperations (11.3%) and 1 readmission (1.6%). No differences were found between groups in any outcome measure. In the 27 patients with an isolated fracture, there were no differences in any outcome measure when treated as an inpatient or outpatient. CONCLUSIONS: We suggest that type I open distal radius fractures could be safely treated surgically >24 hours post-injury without increased risk of infection.

In vitro and in vivo induction of fetal hemoglobin with a reversible and selective DNMT1 inhibitor.

Pharmacological induction of fetal hemoglobin (HbF) expression is an effective therapeutic strategy for the management of beta-hemoglobinopathies such as sickle cell disease. DNA methyltransferase (DNMT) inhibitors 5-azacytidine (5-aza) and 5-aza-2'-deoxycytidine (decitabine) have been shown to induce fetal hemoglobin expression in both preclinical models and clinical studies, but are not currently approved for the management of hemoglobinopathies. We report here the discovery of a novel class of orally bioavailable DNMT1-selective inhibitors as exemplified by GSK3482364. This molecule potently inhibits the methyltransferase activity of DNMT1, but not DNMT family members DNMT3A or DNMT3B. In contrast with cytidine analog DNMT inhibitors, the DNMT1 inhibitory mechanism of GSK3482364 does not require DNA incorporation and is reversible. In cultured human erythroid progenitor cells (EPCs), GSK3482364 decreased overall DNA methylation resulting in de-repression of the gamma globin genes HBG1 and HBG2 and increased HbF expression. In a transgenic mouse model of sickle cell disease, orally administered GSK3482364 caused significant increases in both HbF levels and in the percentage HbF-expressing erythrocytes, with good overall tolerability. We conclude that in these preclinical models, selective, reversible inhibition of DNMT1 is sufficient for the induction of HbF, and is well-tolerated. We anticipate that GSK3482364 will be a useful tool molecule for the further study of selective DNMT1 inhibition both in vitro and in vivo.

Non invasive imaging assessment of the biodistribution of GSK2849330, an ADCC and CDC optimized anti HER3 mAb, and its role in tumor macrophage recruitment in human tumor-bearing mice.

The purpose of this work was to use various molecular imaging techniques to non-invasively assess GSK2849330 (anti HER3 ADCC and CDC enhanced 'AccretaMab' monoclonal antibody) pharmacokinetics and pharmacodynamics in human xenograft tumor-bearing mice. Immuno-PET biodistribution imaging of radiolabeled 89Zr-GSK2849330 was assessed in mice with HER3 negative (MIA-PaCa-2) and positive (CHL-1) human xenograft tumors. Dose dependency of GSK2849330 disposition was assessed using varying doses of unlabeled GSK2849330 co-injected with 89Zr-GSK2849330. In-vivo NIRF optical imaging and ex-vivo confocal microscopy were used to assess the biodistribution of GSK2849330 and the HER3 receptor occupancy in HER3 positive xenograft tumors (BxPC3, and CHL-1). Ferumoxytol (USPIO) contrast-enhanced MRI was used to investigate the effects of GSK2849330 on tumor macrophage content in CHL-1 xenograft bearing mice. Immuno-PET imaging was used to monitor the whole body drug biodistribution and CHL-1 xenograft tumor uptake up to 144 hours post injection of 89Zr-GSK2849330. Both hepatic and tumor uptake were dose dependent and saturable. The optical imaging data in the BxPC3 xenograft tumor confirmed the tumor dose response finding in the Immuno-PET study. Confocal microscopy showed a distinguished cytoplasmic punctate staining pattern within individual CHL-1 cells. GSK2849330 inhibited tumor growth and this was associated with a significant decrease in MRI signal to noise ratio after USPIO injection and with a significant increase in tumor macrophages as confirmed by a quantitative immunohistochemistry analysis. By providing both dose response and time course data from both 89Zr and fluorescently labeled GSK2849330, complementary imaging studies were used to characterize GSK2849330 biodistribution and tumor uptake in vivo. Ferumoxytol-enhanced MRI was used to monitor aspects of the immune system response to GSK2849330. Together these approaches potentially provide clinically translatable, non-invasive techniques to support dose optimization, and assess immune activation and anti-tumor responses.

High-Speed Laser Image Analysis of Plume Angles for Pressurised Metered Dose Inhalers: The Effect of Nozzle Geometry.

The aim of this study is to investigate aerosol plume geometries of pressurised metered dose inhalers (pMDIs) using a high-speed laser image system with different actuator nozzle materials and designs. Actuators made from aluminium, PET and PTFE were manufactured with four different nozzle designs: cone, flat, curved cone and curved flat. Plume angles and spans generated using the designed actuator nozzles with four solution-based pMDI formulations were imaged using Oxford Lasers EnVision system and analysed using EnVision Patternate software. Reduced plume angles for all actuator materials and nozzle designs were observed with pMDI formulations containing drug with high co-solvent concentration (ethanol) due to the reduced vapour pressure. Significantly higher plume angles were observed with the PTFE flat nozzle across all formulations, which could be a result of the nozzle geometry and material's hydrophobicity. The plume geometry of pMDI aerosols can be influenced by the vapour pressure of the formulation, nozzle geometries and actuator material physiochemical properties.

The Effect of Active Pharmaceutical Ingredients on Aerosol Electrostatic Charges from Pressurized Metered Dose Inhalers.

PURPOSE: This study investigated the effect of different active pharmaceutical ingredients (API) on aerosol electrostatic charges and aerosol performances for pressurized metered dose inhalers (pMDIs), using both insulating and conducting actuators. METHODS: Five solution-based pMDIs containing different API ingredients including: beclomethasone dipropionate (BDP), budesonide (BUD), flunisolide (FS), salbutamol base (SB) and ipratropium bromide (IPBr) were prepared using pressure filling technique. Actuator blocks made from nylon, polytetrafluoroethylene (PTFE) and aluminium were manufactured with 0.3 mm nominal orifice diameter and cone nozzle shape. Aerosol electrostatics for each pMDI formulation and actuator were evaluated using the electrical low-pressure impactor (ELPI) and drug depositions were analysed using high performance liquid chromatography (HPLC). RESULTS: All three actuator materials showed the same net charge trend across the five active drug ingredients, with BDP, BUD and FS showing positive net charges for both nylon and PTFE actuators, respectively. While SB and IPBr had significantly negative net charges across the three different actuators, which correlates to the ionic functional groups present on the drug molecule structures. CONCLUSIONS: The API present in a pMDI has a dominant effect on the electrostatic properties of the formulation, overcoming the charge effect arising from the actuator materials. Results have shown that the electrostatic charges for a solution-based pMDI could be related to the interactions of the chemical ingredients and change in the work function for the overall formulation.

The effect of actuator nozzle designs on the electrostatic charge generated in pressurised metered dose inhaler aerosols.

PURPOSE: To investigate the influence of different actuator nozzle designs on aerosol electrostatic charges and aerosol performances for pressurised metered dose inhalers (pMDIs). METHODS: Four actuator nozzle designs (flat, curved flat, cone and curved cone) were manufactured using insulating thermoplastics (PET and PTFE) and conducting metal (aluminium) materials. Aerosol electrostatic profiles of solution pMDI formulations containing propellant HFA 134a with different ethanol concentration and/or model drug beclomethasone dipropionate (BDP) were studied using a modified electrical low-pressure impactor (ELPI) for all actuator designs and materials. The mass of the deposited drug was analysed using high performance liquid chromatography (HPLC). RESULTS: Both curved nozzle designs for insulating PET and PTFE actuators significantly influenced aerosol electrostatics and aerosol performance compared with conducting aluminium actuator, where reversed charge polarity and higher throat deposition were observed with pMDI formulation containing BDP. Results are likely due to the changes in plume geometry caused by the curved edge nozzle designs and the bipolar charging nature of insulating materials. CONCLUSIONS: This study demonstrated that actuator nozzle designs could significantly influence the electrostatic charges profiles and aerosol drug deposition pattern of pMDI aerosols, especially when using insulating thermoplastic materials where bipolar charging is more dominant.

Experimental investigation into the impact of a liquid droplet onto a granular bed using three-dimensional, time-resolved, particle tracking.

An experimental investigation into the interaction that occurs between an impacting water droplet and a granular bed of loose graded sand has been carried out. High-speed imaging, three-dimensional time-resolved particle tracking, and photogrammetric surface profiling have been used to examine individual impact events. The focus of the study is the quantification and trajectory analysis of the particles ejected from the sand bed, along with measurement of the change in bed morphology. The results from the experiments have detailed two distinct mechanisms of particle ejection: the ejection of water-encapsulated particles from the edge of the wetted region and the ejection of dry sand from the periphery of the impact crater. That the process occurs by these two distinct mechanisms has hitherto been unobserved. Presented in the paper are distributions of the particle ejection velocities, angles, and transport distances for both mechanisms. The ejected water-encapsulated particles, which are few in number, are characterized by low ejection angles and high ejection velocities, leading to large transport distances; the ejected dry particles, which are much greater in number, are characterized by high ejection angles and low velocities, leading to lower transport distances. From the particle ejection data, the momentum of the individual ballistic sand particles has been calculated; it was found that only 2% of the water-droplet momentum at impact is transferred to the ballistic sand particles. In addition to the particle tracking, surface profiling of the granular bed postimpact has provided detailed information on its morphology; these data have demonstrated the consistent nature of the craters produced by the impact and suggest that particle agglomerations released from their edges make up about twice the number of particles involved in ballistic ejection. It is estimated that, overall, about 4% of the water-droplet momentum is taken up in particle movement.

The influence of actuator materials and nozzle designs on electrostatic charge of pressurised metered dose inhaler (pMDI) formulations.

PURPOSE: To investigate the influence of different actuator materials and nozzle designs on the electrostatic charge properties of a series of solution metered dose inhaler (pMDI) aerosols. METHODS: Actuators were manufactured with flat and cone nozzle designs using five different materials from the triboelectric series (Nylon, Polyethylene terephthalate, Polyethylene-High density, Polypropylene copolymer and Polytetrafluoroethylene). The electrostatic charge profiles of pMDI containing beclomethasone dipropionate (BDP) as model drug in HFA-134a propellant, with different concentrations of ethanol were studied. Electrostatic measurements were taken using a modified electrical low-pressure impactor (ELPI) and the deposited drug mass assayed chemically using HPLC. RESULTS: The charge profiles of HFA 134a alone have shown strong electronegativity with all actuator materials and nozzle designs, at an average of -1531.34 pC ± 377.34. The presence of co-solvent ethanol significantly reduced the negative charge magnitude. BDP reduced the suppressing effect of ethanol on the negative charging of the propellant. For all tested formulations, the flat nozzle design showed no significant differences in net charge between different actuator materials, whereas the charge profiles of cone designs followed the triboelectric series. CONCLUSION: The electrostatic charging profiles from a solution pMDI containing BDP and ethanol can be significantly influenced by the actuator material, nozzle design and formulation components. Ethanol concentration appears to have the most significant impact. Furthermore, BDP interactions with ethanol and HFA have an influence on the electrostatic charge of aerosols. By choosing different combinations of actuator materials and orifice design, the fine particle fractions of formulations can be altered.

Quantification of pharmaceuticals, personal care products, and perfluoroalkyl substances in the marine sediments of Puget Sound, Washington, USA.

Concentrations of 119 pharmaceuticals and personal care products (PPCPs) and 13 perfluoroalkyl substances (PFASs) in marine sediments measured throughout Puget Sound (n = 10) and Bellingham Bay (n = 30), Washington, USA, are reported. These data are among the first measurements of PPCPs and PFASs in marine sediments from the Pacific Northwest and provide a comparison to previous measurements of these chemicals in influent, effluent, and biosolids from municipal wastewater treatment plants throughout the region. The concentrations of both PPCPs and PFASs in sediments from Puget Sound and Bellingham Bay ranged from very low to non-detectable for most compounds. Only 14 of the 119 PPCPs and 3 of 13 PFASs were quantifiable in sediments. Diphenhydramine (an antihistamine) was most frequently detected (87.5% of samples), with a maximum concentration of 4.81 ng/g dry weight and an estimated mean detected concentration of 1.68 ng/g. Triclocarban (an antibacterial) was detected in 35.0% of the samples, with a maximum concentration of 16.6 ng/g dry weight. Perfluoroalkyl substances were detected in 2.5% of analyses. Perfluorobutanoate, perfluorooctane sulfonate, and perfluorooctane sulfonamide were detected in 7, 5, and 1 sample(s) each, respectively, with the highest concentrations observed for perfluorooctane sulfonate (1.5 ng/g). Detected concentrations were often highest within the industrial harbor in Bellingham Bay and near the cities of Seattle and Bremerton. Environ Toxicol Chem 2013;32:1701-1710. © 2013 SETAC.

In vitro and in vivo characterization of a novel soluble epoxide hydrolase inhibitor.

Soluble epoxide hydrolase (sEH, EPHX2) metabolizes eicosanoid epoxides, including epoxyeicosatrienoic acids (EETs) to the corresponding dihydroxyeicosatrienoic acids (DHETs), and leukotoxin (LTX) to leukotoxin diol (LTX diol). EETs, endothelium-derived hyperpolarizing factors, exhibit potentially beneficial properties, including anti-inflammatory effects and vasodilation. A novel, potent, selective inhibitor of recombinant human, rat and mouse sEH, GSK2256294A, exhibited potent cell-based activity, a concentration-dependent inhibition of the conversion of 14,15-EET to 14,15-DHET in human, rat and mouse whole blood in vitro, and a dose-dependent increase in the LTX/LTX diol ratio in rat plasma following oral administration. Mice receiving 10 days of cigarette smoke exposure concomitant with oral administration of GSK2256294A exhibited significant, dose-dependent reductions in pulmonary leukocytes and keratinocyte chemoattractant (KC, CXCL1) levels. Mice receiving oral administration of GSK2256294A following 10 days of cigarette smoke exposure exhibited significant reductions in pulmonary leukocytes compared to vehicle-treated mice. These data indicate that GSK2256294A attenuates cigarette smoke-induced inflammation by both inhibiting its initiation and/or maintenance and promoting its resolution. Collectively, these data indicate that GSK2256294A would be an appropriate agent to evaluate the role of sEH in clinical studies, for example in diseases where cigarette smoke is a risk factor, such as chronic obstructive pulmonary disease (COPD) and cardiovascular disease.

T cell depletion protects against alveolar destruction due to chronic cigarette smoke exposure in mice.

The role of T cells in chronic obstructive pulmonary disease (COPD) is not well understood. We have previously demonstrated that chronic cigarette smoke exposure can lead to the accumulation of CD4(+) and CD8(+) T cells in the alveolar airspaces in a mouse model of COPD, implicating these cells in disease pathogenesis. However, whether specific inhibition of T cell responses represents a therapeutic strategy has not been fully investigated. In this study inhibition of T cell responses through specific depleting antibodies, or the T cell immunosuppressant drug cyclosporin A, prevented airspace enlargement and neutrophil infiltration in a mouse model of chronic cigarette smoke exposure. Furthermore, individual inhibition of either CD4(+) T helper or CD8(+) T cytotoxic cells prevented airspace enlargement to a similar degree, implicating both T cell subsets as critical mediators of the adaptive immune response induced by cigarette smoke exposure. Importantly, T cell depletion resulted in significantly decreased levels of the Th17-associated cytokine IL-17A, and of caspase 3 and caspase 7 gene expression and activity, induced by cigarette smoke exposure. Finally, inhibition of T cell responses in a therapeutic manner also inhibited cigarette smoke-induced airspace enlargement, IL-17A expression, and neutrophil influx in mice. Together these data demonstrate for the first time that therapeutic inhibition of T cell responses may be efficacious in the treatment of COPD. Given that broad immunosuppression may be undesirable in COPD patients, this study provides proof-of-concept for more targeted approaches to inhibiting the role of T cells in emphysema development.

Revision of sediment quality triad indicators in Puget Sound (Washington, USA): I. a Sediment Chemistry Index and targets for mixtures of toxicants.

The Washington State Department of Ecology annually conducts sediment quality monitoring in Puget Sound as a component of the Puget Sound Ecosystem Monitoring Program. Sediment samples are analyzed to determine the concentrations of about 170 chemical and physical variables. A Sediment Chemistry Index (SCI) was derived using the State of Washington Sediment Management Standards to account for the presence and concentrations of mixtures of toxicants. Mean Sediment Quality Standard quotients (mSQSq) were calculated as the basis for the SCI and compared to the incidence and degree of toxicity in laboratory tests and to metrics of the diversity and abundance of resident benthic assemblages in a database consisting of as many as 664 samples. These data were evaluated with co-occurrence analyses to identify "cut points" (i.e., thresholds) in the index below which the frequency and magnitude of biological effects were relatively low and above which they occurred with increasing frequency or magnitude. Iterative trials of different sets of cut points established the final cut points in mSQSq of 0.1, 0.3, and 0.5. They defined 4 ranges in chemical exposure: Minimum (<0.1), Low (0.1- < 0.3), Moderate (0.3- < 0.5), and Maximum (≥0.5). Across these 4 exposure ranges both the incidence and magnitude of toxicity in some laboratory tests increased, the abundance of most stress-sensitive benthic taxa decreased, and the abundance of most stress-tolerant taxa increased. The mSQSq cut point of 0.1 appears to be the target value for protection of benthic resources, the value below which the probability and magnitude of adverse effects either in the laboratory or the field are the lowest. The mSQSq values are rescaled from 0 to 100 to form the SCI, used by the Puget Sound Partnership and environmental managers as a Dashboard Indicator, with biologically relevant targets selected to monitor ecosystem recovery.

Toxicity of sediment pore water in Puget Sound (Washington, USA): a review of spatial status and temporal trends.

Data from toxicity tests of the pore water extracted from Puget Sound sediments were compiled from surveys conducted from 1997 to 2009. Tests were performed on 664 samples collected throughout all of the eight monitoring regions in the Sound, an area encompassing 2,294.1 km(2). Tests were performed with the gametes of the Pacific purple sea urchin, Strongylocentrotus purpuratus, to measure percent fertilization success as an indicator of relative sediment quality. Data were evaluated to determine the incidence, degree of response, geographic patterns, spatial extent, and temporal changes in toxicity. This is the first survey of this kind and magnitude in Puget Sound. In the initial round of surveys of the eight regions, 40 of 381 samples were toxic for an incidence of 10.5 %. Stations classified as toxic represented an estimated total of 107.1 km(2), equivalent to 4.7 % of the total area. Percent sea urchin fertilization ranged from >100 % of the nontoxic, negative controls to 0 %. Toxicity was most prevalent and pervasive in the industrialized harbors and lowest in the deep basins. Conditions were intermediate in deep-water passages, urban bays, and rural bays. A second round of testing in four regions and three selected urban bays was completed 5-10 years following the first round. The incidence and spatial extent of toxicity decreased in two of the regions and two of the bays and increased in the other two regions and the third bay; however, only the latter change was statistically significant. Both the incidence and spatial extent of toxicity were lower in the Sound than in most other US estuaries and marine bays.

Airway infiltration of CD4+ CCR6+ Th17 type cells associated with chronic cigarette smoke induced airspace enlargement.

Recently, patients with tobacco smoke induced emphysema have been shown to exhibit classical signs of T cell mediated autoimmunity characterized by autoantibody production and Th1 type responses. As the recently described Th17 type subset has been found to play a role in the pathogenesis of a number of autoimmune diseases previously considered to be Th1 driven, we sought to examine whether a Th17 type response was associated with airspace enlargement in a murine model of emphysema. Six to eight months exposure of mice to inhalation of mainstream cigarette smoke led to progressive airspace enlargement as defined by morphometric analysis. Flow cytometric analysis of the bronchoalveolar lavage (BAL) from these mice demonstrated a significant increase in the overall number of both CD4+ and CD8+ T cells present. These cells were subsequently examined for skewing towards a Th1, Th2 or Th17 phenotype by intracellular cytokine analysis. Distinct populations of BAL CD4+ T cells were found to express IFN-gamma or IL-17 demonstrating the presence of both a Th1 and Th17 type response. No expression of the Th2 associated cytokine IL-4 was detected. Further analysis of this Th17 subset demonstrated that the majority of cells with this effector phenotype express the chemokine receptor CCR6. Together these data identify a novel T cell subset associated with pulmonary inflammation as a result of cigarette smoke exposure. Given the reported roles of CCR6 and IL-17 in promoting pulmonary inflammation, this subset may play an important role in the pathogenesis of cigarette smoke induced autoimmunity.

RecA-mediated SOS induction requires an extended filament conformation but no ATP hydrolysis.

The Escherichia coli SOS response to DNA damage is modulated by the RecA protein, a recombinase that forms an extended filament on single-stranded DNA and hydrolyzes ATP. The RecA K72R (recA2201) mutation eliminates the ATPase activity of RecA protein. The mutation also limits the capacity of RecA to form long filaments in the presence of ATP. Strains with this mutation do not undergo SOS induction in vivo. We have combined the K72R variant of RecA with another mutation, RecA E38K (recA730). In vitro, the double mutant RecA E38K/K72R (recA730,2201) mimics the K72R mutant protein in that it has no ATPase activity. The double mutant protein will form long extended filaments on ssDNA and facilitate LexA cleavage almost as well as wild-type, and do so in the presence of ATP. Unlike recA K72R, the recA E38K/K72R double mutant promotes SOS induction in vivo after UV treatment. Thus, SOS induction does not require ATP hydrolysis by the RecA protein, but does require formation of extended RecA filaments. The RecA E38K/K72R protein represents an improved reagent for studies of the function of ATP hydrolysis by RecA in vivo and in vitro.

Inhibition of invariant chain processing, antigen-induced proliferative responses, and the development of collagen-induced arthritis and experimental autoimmune encephalomyelitis by a small molecule cysteine protease inhibitor.

Members of the papain family of cysteine proteases (cathepsins) mediate late stage processing of MHC class II-bound invariant chain (Ii), enabling dissociation of Ii, and binding of antigenic peptide to class II molecules. Recognition of cell surface class II/Ag complexes by CD4(+) T cells then leads to T cell activation. Herein, we demonstrate that a pan-active cathepsin inhibitor, SB-331750, attenuated the processing of whole cell Ii p10 to CLIP by Raji cells, and DBA/1, SJL/J, and C57BL/6 splenocytes. In Raji cells and C57BL/6 splenocytes, SB-331750 inhibited class II-associated Ii processing and reduced surface class II/CLIP expression, whereas in SB-331750-treated DBA/1 and SJL/J splenocytes, class II-associated Ii processing intermediates were undetectable. Incubation of lymph node cells/splenocytes from collagen-primed DBA/1 mice and myelin basic protein-primed SJL/J mice with Ag in the presence of SB-331750 resulted in concentration-dependent inhibition of Ag-induced proliferation. In vivo administration of SB-331750 to DBA/1, SJL/J, and C57BL/6 mice inhibited splenocyte processing of whole cell Ii p10 to CLIP. Prophylactic administration of SB-331750 to collagen-immunized/boosted DBA/1 mice delayed the onset and reduced the severity of collagen-induced arthritis (CIA), and reduced paw tissue levels of IL-1beta and TNF-alpha. Similarly, treatment of myelin basic protein-primed SJL/J lymph node cells with SB-331750 delayed the onset and reduced the severity of adoptively transferred experimental autoimmune encephalomyelitis (EAE). Therapeutic administration of SB-331750 reduced the severity of mild/moderate CIA and EAE. These results indicate that pharmacological inhibition of cathepsins attenuates CIA and EAE, potentially via inhibition of Ii processing, and subsequent Ag-induced T cell activation.

Sediment quality triad assessment of an industrialized estuary of the northeastern USA.

The lower Passaic River in northern New Jersey (USA) has been heavily industrialized since the mid-nineteenth century and its shoreline and aquatic habitats degraded or destroyed. Similar to other urban systems, Passaic River sediments, both surface and buried, historically have contained elevated levels of numerous contaminants that may pose risks to ecological receptors and humans. Sediments from 15 stations in the lower Passaic River and 3 reference stations in the Mullica River in southern New Jersey were sampled in 1999 and characterized for chemical contamination, toxicity, and impairment of the benthic community. The objective of this study was to determine the incidence, degree, and nature of degraded surficial sediments in the area to support subsequent plans for restoration of the system. Results demonstrated that Passaic River sediments had concentrations of many organic and inorganic contaminants at levels significantly greater than the reference area and effect-based guidelines. Sediments were toxic to marine amphipods at 11 stations and the benthic assemblages were impaired relative to the reference area at all stations. The weight-of-evidence of this sediment quality triad (SQT) assessment indicates that impacts from multiple contaminants are occurring throughout the lower Passaic River and, that these impacts must be evaluated further and addressed as part of ongoing restoration initiatives for the river.

Evaluating consistency of best professional judgment in the application of a multiple lines of evidence sediment quality triad.

The bioavailability of sediment-associated contaminants is poorly understood. Often, a triad of chemical concentration measurements, laboratory sediment toxicity tests, and benthic infaunal community condition is used to assess whether contaminants are present at levels of ecological concern. Integration of these 3 lines of evidence is typically based on best professional judgment by experts; however, the level of consistency among expert approach and interpretation has not been determined. In this study, we compared the assessments of 6 experts who were independently provided data from 25 California embayment sites and asked to rank the relative condition of each site from best to worst. The experts were also asked to place each site into 1 of 6 predetermined categories of absolute condition. We provided no guidance regarding assessment approach or interpretation of supplied data. The relative ranking of the sites was highly correlated among the experts, with an average correlation coefficient of 0.92. Although the experts' relative rankings were highly correlated, the categorical assessments were much less consistent, with only 1 site out of 25 assigned to the same absolute condition category by all 6 experts. Most of the observed categorical differences were small in magnitude and involved the weighing of different lines of evidence in individual assessment approaches, rather than interpretation of signals within a line of evidence. We attribute categorical differences to the experts' use of individual best professional judgment and consider these differences to be indicative of potential uncertainty in the evaluation of sediment quality. The results of our study suggest that specifying key aspects of the assessment approach a priori and aligning the approach to the study objectives can reduce this uncertainty.

Decreased pCO(2) accumulation by eliminating bicarbonate addition to high cell-density cultures.

High-density perfusion cultivation of mammalian cells can result in elevated bioreactor CO(2) partial pressure (pCO(2)), a condition that can negatively influence growth, metabolism, productivity, and protein glycosylation. For BHK cells in a perfusion culture at 20 x 10(6) cells/mL, the bioreactor pCO(2) exceeded 225 mm Hg with approximate contributions of 25% from cellular respiration, 35% from medium NaHCO(3), and 40% from NaHCO(3) added for pH control. Recognizing the limitations to the practicality of gas sparging for CO(2) removal in perfusion systems, a strategy based on CO(2) reduction at the source was investigated. The NaHCO(3) in the medium was replaced with a MOPS-Histidine buffer, while Na(2)CO(3) replaced NaHCO(3) for pH control. These changes resulted in 63-70% pCO(2) reductions in multiple 15 L perfusion bioreactors, and were reproducible at the manufacturing-scale. Bioreactor pCO(2) values after these modifications were in the 68-85 mm Hg range, pCO(2) reductions consistent with those theoretically expected. Low bioreactor pCO(2) was accompanied by both 68-123% increased growth rates and 58-92% increased specific productivity. Bioreactor pCO(2) reduction and the resulting positive implications for cell growth and productivity were brought about by process changes that were readily implemented and robust. This philosophy of pCO(2) reduction at the source through medium and base modification should be readily applicable to large-scale fed-batch cultivation of mammalian cells.

Evaluation of potential toxicity and bioavailability of chromium in sediments associated with chromite ore processing residue.

Recent studies by researchers at the U.S. Environmental Protection Agency and U.S. Geological Survey have evaluated the toxicity of Cr in freshwater and marine sediments, primarily during laboratory studies in which clean sediments were spiked with Cr. Results of those studies showed that Cr is relatively insoluble and nontoxic when present in the trivalent form, Cr(III), rather than in the more soluble and more toxic hexavalent form, Cr(VI). The studies concluded that Cr toxicity should be low in sediments with measurable concentrations of acid-volatile sulfide (AVS), because AVS is formed only in anoxic sediments and Cr(VI) is thermodynamically unstable under such conditions. The present study evaluates the toxicity and bioavailability of Cr in sediments associated with chromite ore processing residue (COPR). Ten stations were sampled in the Hackensack River (NJ, USA) to represent a wide range of total Cr concentrations (199-3,970 mg/kg) with minimal interference from potentially toxic, co-occurring chemicals. Sediment toxicity was evaluated using two amphipod tests: The 10-d Ampelisca abdita test (survival as endpoint), and the 28-d Leptocheirus plumulosus test (survival and biomass as endpoints). Measurable concentrations of AVS were present at eight stations, and nearly all Cr was present as Cr(III). In addition, results of electron-microprobe analyses showed that most Cr was associated with phases in which Cr has limited bioavailability (i.e., chromite and iron oxide). Sediment toxicity showed no correlation with concentrations of total Cr, and the maximum no-effect concentration for total Cr was estimated as 1,310 mg/ kg. These results indicate that Cr can be present in sediments associated with COPR at highly elevated concentrations without causing sediment toxicity.