Evaluation of BBB permeable nucleolipid (NLDPU): A di-C15-ketalised palmitone appended uridine as neuro-tracer for SPECT.

Despite being in routine for onco-diagnostics for years, the applicability of nucleosidic molecular imaging probes is severely restricted in neurological applications due to their low permeability across blood-brain-barrier (BBB). For extending nucleoside tracers utility for neuro-onco early diagnostics, suitable modification which enhances their BBB permeation needs investigation. Among various modifications, lipidization of nucleosides has been reported to enhance cellular permeability. Extending the concept, the aim was to exemplify the possibility of lipidized nucleosides as potential brain tracer with capability to cross intact BBB and evaluate as metal based neuro-imaging SPECT agent. Uridine based non-lipidic (NSDAU) and di-C15-ketal appended lipidic (NLDPU) ligands were conjugated to chelator, DTPA (DTPA-NSDAU and DTPA-NLDPU) using multi-step chemistry. The ligands were evaluated in parallel for comparative physical and biological parameters. Additionally, effects of enhanced lipophilicity on UV-absorption, acid strength, fluorescence and non-specific protein binding were evaluated. Fluorescence quenching of BSA indicated appreciable interaction of DTPA-NLDPU with protein only above 10 mM without inducing conformational changes. In addition, DTPA-NLDPU was found to be haematocompatible and cytocompatible with low dose-dependent toxicity in HEK-cells. The chelator DTPA was used for 99mTc-complexation for SPECT imaging. Optimized 99mTc-radiolabeling parameters resulted in quantitative (≥97%) labeling with good stability parameters in in-vitro serum and cysteine challenge studies. We demonstrate that the nucleolipid radiotracer (99mTc-DTPA-NLDPU) was successfully able to permeate the BBB with brain uptake of 0.2% ID/g in normal mice as compared to 0.06% ID/g uptake of 99mTc-DTPA-NSDAU at 5 min. Blood kinetics indicate biphasic profile and t1/2(distribution) 46 min for 99mTc-DTPA-NLDPU. The preferential accumulation of 99mTc-DTPA-NLDPU in brain tumor intracranial xenograft indicate the targeting capability of the nucleoside. We conclude that as first-of-its-kind, this work presents the potential of the biocompatible nucleolipidic system for brain targeting and early diagnostics.

Arsenic-containing hydrocarbons: effects on gene expression, epigenetics, and biotransformation in HepG2 cells.

Arsenic-containing hydrocarbons (AsHCs), a subgroup of arsenolipids found in fish and algae, elicit substantial toxic effects in various human cell lines and have a considerable impact on cellular energy levels. The underlying mode of action, however, is still unknown. The present study analyzes the effects of two AsHCs (AsHC 332 and AsHC 360) on the expression of 44 genes covering DNA repair, stress response, cell death, autophagy, and epigenetics via RT-qPCR in human liver (HepG2) cells. Both AsHCs affected the gene expression, but to different extents. After treatment with AsHC 360, flap structure-specific endonuclease 1 (FEN1) as well as xeroderma pigmentosum group A complementing protein (XPA) and (cytosine-5)-methyltransferase 3A (DNMT3A) showed time- and concentration-dependent alterations in gene expression, thereby indicating an impact on genomic stability. In the subsequent analysis of epigenetic markers, within 72 h, neither AsHC 332 nor AsHC 360 showed an impact on the global DNA methylation level, whereas incubation with AsHC 360 increased the global DNA hydroxymethylation level. Analysis of cell extracts and cell media by HPLC-mass spectrometry revealed that both AsHCs were considerably biotransformed. The identified metabolites include not only the respective thioxo-analogs of the two AsHCs, but also several arsenic-containing fatty acids and fatty alcohols, contributing to our knowledge of biotransformation mechanisms of arsenolipids.

Accumulation of mineral oil saturated hydrocarbons (MOSH) in female Fischer 344 rats: Comparison with human data and consequences for risk assessment.

Female Fischer 344 rats were orally exposed to a mixture of mineral oil saturated hydrocarbons (MOSH) of broad molecular mass range at doses of 40, 400 and 4000mg/kg feed. Amounts and compositions of the MOSH were analyzed in liver, spleen, adipose tissue and the carcass after exposure during 30, 60, 90 and 120d as well as after 90d exposure followed by 30d depuration. At 40mg/kg in the feed, after 30d of exposure, 10.9% of the ingested MOSH were recovered from the animal body; after 90d plus 30d depuration it was 3.9%. In liver and spleen, the maximum retention in terms of molecular mass (simulated distillation) was at n-C29; in adipose tissue and carcass it was at n-C15/16. The differentiation between MOSH below and above n-C25 (Class I versus Class II and III oils), used for present regulation, is not supported by the present data on accumulation; structural characteristics seem more pertinent than molecular mass. Concentrations in the tissues increased far less than proportionally with the dose, rendering linear extrapolation to low doses questionable. No steady state was reached after 120d. In fact, comparing with the concentrations in human tissues at the estimated exposure, extrapolation from animal experiments seems to grossly underestimate human internal exposure. Comprehensive two-dimensional gas chromatography (GCxGC) was used to characterize the MOSH residues in the tissues with the aim of identifying the most strongly accumulated types. In the liver and spleen, the highly branched hydrocarbons dominated, whereas in the adipose tissue it was the n-alkanes and species with main n-alkyl moieties. Strong MOSH accumulation is not of concern per se, but the safety at the high concentrations in human tissues needs to be re-evaluated, possibly taking into account also end points other than granuloma formation.

Biophysical determinants for cellular uptake of hydrocarbon-stapled peptide helices.

Hydrocarbon-stapled peptides are a class of bioactive alpha-helical ligands developed to dissect and target protein interactions. While there is consensus that stapled peptides can be effective chemical tools for investigating protein regulation, their broader utility for therapeutic modulation of intracellular interactions remains an active area of study. In particular, the design principles for generating cell-permeable stapled peptides are empiric, yet consistent intracellular access is essential to in vivo application. Here, we used an unbiased statistical approach to determine which biophysical parameters dictate the uptake of stapled-peptide libraries. We found that staple placement at the amphipathic boundary combined with optimal hydrophobic and helical content are the key drivers of cellular uptake, whereas excess hydrophobicity and positive charge at isolated amino acid positions can trigger membrane lysis at elevated peptide dosing. Our results provide a design roadmap for maximizing the potential to generate cell-permeable stapled peptides with on-mechanism cellular activity.

Quantitative cancer risk assessment for occupational exposures to asphalt fumes during built-up roofing asphalt (BURA) operations.

The International Agency for Research on Cancer qualitatively characterized occupational exposure to oxidized bitumen emissions during roofing as probably carcinogenic to humans (Group 2A). We examine chemistry, exposure, epidemiology and animal toxicity data to explore quantitative risks for roofing workers applying built-up roofing asphalt (BURA). Epidemiology studies do not consistently report elevated risks, and generally do not have sufficient exposure information or adequately control for confounders, precluding their use for dose-response analysis. Dermal carcinogenicity bioassays using mice report increased tumor incidence with single high doses. In order to quantify potential cancer risks, we develop time-to-tumor model methods [consistent with U.S. Environmental Protection Agency (EPA) dose-response analysis and mixtures guidelines] using the dose-time-response shape of concurrent exposures to benzo[a]pyrene (B[a]P) as concurrent controls (which had several exposure levels) to infer presumed parallel dose-time-response curves for BURA-fume condensate. We compare EPA relative potency factor approaches, based on observed relative potency of BURA to B[a]P in similar experiments, and direct observation of the inferred BURA dose-time-response (scaled to humans) as means for characterizing a dermal unit risk factor. We apply similar approaches to limited data on asphalt-fume inhalation and respiratory cancers in rats. We also develop a method for adjusting potency estimates for asphalts that vary in composition using measured fluorescence. Overall, the various methods indicate that cancer risks to roofers from both dermal and inhalation exposure to BURA are within a range typically deemed acceptable within regulatory frameworks. The approaches developed may be useful in assessing carcinogenic potency of other complex mixtures of polycyclic aromatic compounds.

Cellular Uptake and Ultrastructural Localization Underlie the Pro-apoptotic Activity of a Hydrocarbon-stapled BIM BH3 Peptide.

Hydrocarbon stapling has been applied to restore and stabilize the α-helical structure of bioactive peptides for biochemical, structural, cellular, and in vivo studies. The peptide sequence, in addition to the composition and location of the installed staple, can dramatically influence the properties of stapled peptides. As a result, constructs that appear similar can have distinct functions and utilities. Here, we perform a side-by-side comparison of stapled peptides modeled after the pro-apoptotic BIM BH3 helix to highlight these principles. We confirm that replacing a salt-bridge with an i, i + 4 hydrocarbon staple does not impair target binding affinity and instead can yield a biologically and pharmacologically enhanced α-helical peptide ligand. Importantly, we demonstrate by electron microscopy that the pro-apoptotic activity of a stapled BIM BH3 helix correlates with its capacity to achieve cellular uptake without membrane disruption and accumulate at the organellar site of mechanistic activity.

Role of entrained droplet oil on the bioavailability of petroleum substances in aqueous exposures.

Bioavailability of petroleum substances is a complex issue that is affected by substance composition, the physicochemical properties of the individual constituents, and the exposure preparation system. The present study applies mechanistic fate and effects models to characterize the role of droplet oil on dissolved exposure and predicted effects from both neat and weathered crude oils, and refined fuel oils. The main effect from droplet oil is input of additional dissolved hydrocarbons to the exposure system following preparation of the initial stock solution. Toxicity was characterized using toxic units (TU) and shows that replenishment of bioavailable hydrocarbons by droplets in toxicity tests with low droplet content (e.g., <1mg/L) is negligible, consistent with typical exposure conditions following open ocean oil spills. Further, the use of volumetric exposure metrics (e.g., mg/L) introduces considerable variability and the bioavailability-based metrics (e.g., TUs) provide a more consistent basis for understanding oil toxicity data.

In vitro toxicological characterisation of three arsenic-containing hydrocarbons.

Arsenic-containing hydrocarbons are one group of fat-soluble organic arsenic compounds (arsenolipids) found in marine fish and other seafood. A risk assessment of arsenolipids is urgently needed, but has not been possible because of the total lack of toxicological data. In this study the cellular toxicity of three arsenic-containing hydrocarbons was investigated in cultured human bladder (UROtsa) and liver (HepG2) cells. Cytotoxicity of the arsenic-containing hydrocarbons was comparable to that of arsenite, which was applied as the toxic reference arsenical. A large cellular accumulation of arsenic, as measured by ICP-MS/MS, was observed after incubation of both cell lines with the arsenolipids. Moreover, the toxic mode of action shown by the three arsenic-containing hydrocarbons seemed to differ from that observed for arsenite. Evidence suggests that the high cytotoxic potential of the lipophilic arsenicals results from a decrease in the cellular energy level. This first in vitro based risk assessment cannot exclude a risk to human health related to the presence of arsenolipids in seafood, and indicates the urgent need for further toxicity studies in experimental animals to fully assess this possible risk.

Bioaccumulation of petroleum hydrocarbons in fiddler crabs (Uca minax) exposed to weathered MC-252 crude oil alone and in mixture with an oil dispersant.

The Deepwater Horizon accident in the Gulf of Mexico resulted in a sustained release of crude oil, and weathered oil was reported to have washed onto shorelines and marshes along the Gulf coast. One strategy to minimize effects of tarballs, slicks, and oil sheen, and subsequent risk to nearshore ecosystem resources was to use oil dispersants (primarily Corexit® 9500) at offshore surface and deepwater locations. Data have been generated reporting how Corexit® 9500 and other dispersants may alter the acute toxicity of crude oil (Louisiana sweet crude) to marine organisms. However, it remains unknown how oil dispersants may influence bioaccumulation of petroleum hydrocarbons in nearshore crustaceans. We compare bioaccumulation of petroleum hydrocarbons in fiddler crabs (Uca minax) from exposures to the water accommodated fraction (WAF) of weathered Mississippi Canyon 252 oil (~30 d post spill) and chemically-enhanced WAF when mixed with Corexit® EC9500A. Whole body total petroleum hydrocarbon (TPH) concentrations were greater than background for both treatments after 6h of exposure and reached steady state at 96 h. The modeled TPH uptake rate was greater for crabs in the oil only treatment (k(u)=2.51 mL/g/h vs. 0.76 mL/g/h). Furthermore, during the uptake phase TPH patterns in tissues varied between oil only and oil+dispersant treatments. Steady state bioaccumulation factors (BAFs) were 19.0 mL/g and 14.1 mL/g for the oil only and oil+Corexit treatments, respectively. These results suggest that the toxicokinetic mechanisms of oil may be dependent on oil dispersion (e.g., smaller droplet sizes). The results also indicate that multiple processes and functional roles of species should be considered for understanding how dispersants influence bioavailability of petroleum hydrocarbons.

Phytoassessment of a waste engine oil-polluted soil exposed to two different intervals of monitored natural attenuation using African yam bean (Sphenostylis stenocarpa).

The present study comparatively investigated the phytotoxic effects of waste engine oil (WEO)-polluted soil exposed to monitored natural attenuation up to 5 and 14 months respectively. Soil was previously polluted with WEO at 0, 1, 2.5, 5 and 10% w/w oil in soil. Although, there was significant reduction in heavy metal concentration of soil as well as total hydrocarbon contents, performance of Sphenostylis stenocarpa was greatly retarded when sown at 5 months after pollution (MAP), with death of all seedlings except in the control. However, growth and yield performances were significantly (p > 0.05) enhanced at 14 MAP. Computation of hazard quotient showed that ecological risk factor initially posed by the presence of heavy metals in the soil at 5 MAP was significantly (p > 0.05) reduced to safe levels at 14 MAP.

Evaluation and comparison of urinary metabolic biomarkers of exposure for the jet fuel JP-8.

A study of workers exposed to jet fuel propellant 8 (JP-8) was conducted at U.S. Air Force bases and included the evaluation of three biomarkers of exposure: S-benzylmercapturic acid (BMA), S-phenylmercapturic acid (PMA), and (2-methoxyethoxy)acetic acid (MEAA). Postshift urine specimens were collected from various personnel categorized as high (n = 98), moderate (n = 38) and low (n = 61) JP-8 exposure based on work activities. BMA and PMA urinary levels were determined by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), and MEAA urinary levels were determined by gas chromatography-mass spectrometry (GC-MS). The numbers of samples determined as positive for the presence of the BMA biomarker (above the test method's limit of detection [LOD = 0.5 ng/ml]) were 96 (98.0%), 37 (97.4%), and 58 (95.1%) for the high, moderate, and low (control) exposure workgroup categories, respectively. The numbers of samples determined as positive for the presence of the PMA biomarker (LOD = 0.5 ng/ml) were 33 (33.7%), 9 (23.7%), and 12 (19.7%) for the high, moderate, and low exposure categories. The numbers of samples determined as positive for the presence of the MEAA biomarker (LOD = 0.1 µ g/ml) were 92 (93.4%), 13 (34.2%), and 2 (3.3%) for the high, moderate, and low exposure categories. Statistical analysis of the mean levels of the analytes demonstrated MEAA to be the most accurate or appropriate biomarker for JP-8 exposure using urinary concentrations either adjusted or not adjusted for creatinine; mean levels of BMA and PMA were not statistically significant between workgroup categories after adjusting for creatinine.

A quantitative method for estimating dermal benzene absorption from benzene-containing hydrocarbon liquids.

This study examines a method for estimating the dermal absorption of benzene contained in hydrocarbon liquids that contact the skin. This method applies to crude oil, gasoline, organic solvents, penetrants, and oils. The flux of benzene through occluded skin as a function of the percent vol/vol benzene in the liquid is derived by fitting a curve to experimental data; the function is supralinear at benzene concentrations < or = 5% vol/vol. When a liquid other than pure benzene is on nonoccluded skin, benzene may preferentially evaporate from the liquid, which thereby decreases the benzene flux. We present a time-averaging method here for estimating the reduced dermal flux during evaporation. Example calculations are presented for benzene at 2% vol/vol in gasoline, and for benzene at 0.1% vol/vol in a less volatile liquid. We also discuss other factors affecting dermal absorption.

Urinary metabolites of polycyclic aromatic hydrocarbons in workers exposed to vapours and aerosols of bitumen.

Urinary hydroxylated metabolites of polycyclic aromatic hydrocarbons (PAH) were investigated as potential biomarkers of bitumen exposure in a cross-shift study in 317 exposed and 117 non-exposed workers. Personal measurements of the airborne concentration of vapours and aerosols of bitumen during a working shift were weakly associated with post-shift concentrations of 1-hydroxypyrene (1-OHP) and 1-, 2+9-, 3- and 4-hydroxyphenanthrenes (further referred to their sum as OHPHE), but not 1- and 2-hydroxynaphthalene (OHNA). Smoking showed a strong influence on the metabolite concentrations, in particular on OHNA. Pre-shift concentrations of 1-OHP and OHPHE did not differ between the study groups (P = 0.16 and P = 0.89, respectively). During shift, PAH metabolite concentrations increased in exposed workers and non-exposed smokers. Statistical modelling of post-shift concentrations revealed a small increase in 1-OHP by a factor of 1.02 per 1 mg/m(3) bitumen (P = 0.02) and 1.04 for OHPHE (P < 0.001). A group difference was observed that was diminished in non-smokers. Exposed non-smokers had a median post-shift 1-OHP concentration of 0.42 µg/l, and non-smoking referents 0.13 µg/l. Although post-shift concentrations of 1-OHP and OHPHE were slightly higher than those in the general population, they were much lower than in coke-oven workers. The small content of PAHs in vapours and aerosols of bitumen, the increasing use of additives to asphalt mixtures, the strong impact of smoking and their weak association with airborne bitumen limit the use of PAH metabolites as specific biomarkers of bitumen exposure.

The effect of organism density on bioaccumulation of contaminants from sediment in three aquatic test species: a case for standardizing to sediment organic carbon.

Laboratory methods for measuring bioaccumulation of organic contaminants from sediment into aquatic organisms continue to improve, but some aspects are still in need of standardization. From a review of published methods, we noted that the loading density of organisms was determined inconsistently and was primarily based on either sediment volume or total organic carbon (TOC). The rationale mainly expressed for standardizing to TOC was to minimize the depletion of sediment contaminants. However, even when density was standardized to TOC, the relative amount of TOC provided (i.e., ratio of TOC to organism dry weight [dw]) was highly variable. In this study, we examined the effect of organism density (standardized to sediment TOC or volume) on bioaccumulation in three freshwater organisms. The oligochaete Lumbriculus variegatus, mayfly nymph Hexagenia spp., and fathead minnow Pimephales promelas were exposed for 28 days to two field-contaminated sediments that varied in concentration of PCBs and TOC. Densities tested were 50:1 and 27:1 ratios of TOC to organism dw and 140 ml sediment/g wet weight (ww) biomass, yielding low to high organism densities. Bioaccumulation in Hexagenia spp. was significantly higher at the lowest organism density compared with the highest organism density when exposed to site 2 sediment (1.1% TOC) but only with tissue concentrations expressed on a ww basis. Otherwise, there was no significant effect of density on bioaccumulation in organisms exposed to sediments from site 1 (12% TOC) or site 2. Survival of Hexagenia spp. was adversely affected at the highest organism density when the relative amount of TOC was low. The results of this study support the recommendation of standardizing organism density relative to a particular amount of TOC for invertebrate species. A 27:1 ratio of TOC:organism dw was selected as a standard organism density for a new bioaccumulation method because survival, growth, and bioaccumulation were not impacted relative to a 50:1 ratio, and less sediment was required. This density is recommended as an appropriate ratio for sediment bioaccumulation assessments in general.

Single and joint effects of petroleum hydrocarbons and cadmium on the polychaete Perinereis aibuhitensis Grube.

Using the concentration gradient and combined pollutant exposure method, the single and joint effects of petroleum hydrocarbons (PHCs) and cadmium (Cd) on polychaete Perinereis aibuhitensis Grube, an ecologically keystone species in estuarine and coastal environment, have been investigated. The results indicate that the toxicity of PHCs to P. aibuhitensis is stronger than that of Cd to the organism. There are positive correlations between the mortality of worms and the exposed concentration of single Cd or PHCs in solution. Similarly, the accumulation of Cd or PHCs in worms increased with increasing Cd- or PHC-exposed concentrations. All the correlation relationships can be described using unitary quadratic equations (Y or Z = aX2 + bX + c). It is calculated, on the basis of these expressions, that the median lethal dose (LC50) of P. aibuhitensis exposed to a single Cd or PHCs is 793.4-13567.3 and 28.0-119.9 microg/L, respectively. The exposed time has some stimulative effect on the two pollutants and on the mortality of the worms. Thus, even a low concentration of a single Cd or PHCs may have strong toxic effects on the worms when the exposed time becomes longer. The accumulation of Cd or PHCs in worms differs with an increase in exposure time at the given exposed concentration of a single Cd or PHCs. Noticeably, the accumulation of PHCs in worms decreases with an increase in exposure time at the given high concentration of PHCs in solution. The joint effect of PHCs and Cd on P. aibuhitensis is very complicated and changes with the exposed concentrations of the two pollutants. At the given concentration of PHCs, the joint toxicity of the two pollutants on the worms changes from synergism to antagonism with an increase in Cd concentration. The accumulation of Cd in the worms significantly decreases with the addition of PHCs to exposure solution.

JP-8 jet fuel can promote auditory impairment resulting from subsequent noise exposure in rats.

We report on the transient and persistent effects of JP-8 jet fuel exposure on auditory function in rats. JP-8 has become the standard jet fuel utilized in the United States and North Atlantic Treaty Organization countries for military use and it is closely related to Jet A fuel, which is used in U.S. domestic aviation. Rats received JP-8 fuel (1000 mg/m(3)) by nose-only inhalation for 4 h and half of them were immediately subjected to an octave band of noise ranging between 97 and 105 dB in different experiments. The noise by itself produces a small, but permanent auditory impairment. The current permissible exposure level for JP-8 is 350 mg/m(3). Additionally, a positive control group received only noise exposure, and a fourth group consisted of untreated control subjects. Exposures occurred either on 1 day or repeatedly on 5 successive days. Impairments in auditory function were assessed using distortion product otoacoustic emissions and compound action potential testing. In other rats, tissues were harvested following JP-8 exposure for assessment of hydrocarbon levels or glutathione (GSH) levels. A single JP-8 exposure by itself at 1000 mg/m(3) did not disrupt auditory function. However, exposure to JP-8 and noise produced an additive disruption in outer hair cell function. Repeated 5-day JP-8 exposure at 1000 mg/m(3) for 4 h produced impairment of outer hair cell function that was most evident at the first postexposure assessment time. Partial though not complete recovery was observed over a 4-week postexposure period. The adverse effects of repeated JP-8 exposures on auditory function were inconsistent, but combined treatment with JP-8 + noise yielded greater impairment of auditory function, and hair cell loss than did noise by itself. Qualitative comparison of outer hair cell loss suggests an increase in outer hair cell death among rats treated with JP-8 + noise for 5 days as compared to noise alone. In most instances, hydrocarbon constituents of the fuel were largely eliminated in all tissues by 1-h postexposure with the exception of fat. Finally, JP-8 exposure did result in a significant depletion of total GSH that was observable in liver with a nonsignificant trend toward depletion in the brain and lung raising the possibility that the promotion of noise-induced hearing loss by JP-8 might have resulted from oxidative stress.

Dermal exposure to jet fuel JP-8 significantly contributes to the production of urinary naphthols in fuel-cell maintenance workers.

Jet propulsion fuel 8 (JP-8) is the major jet fuel used worldwide and has been recognized as a major source of chemical exposure, both inhalation and dermal, for fuel-cell maintenance workers. We investigated the contributions of dermal and inhalation exposure to JP-8 to the total body dose of U.S. Air Force fuel-cell maintenance workers using naphthalene as a surrogate for JP-8 exposure. Dermal, breathing zone, and exhaled breath measurements of naphthalene were obtained using tape-strip sampling, passive monitoring, and glass bulbs, respectively. Levels of urinary 1- and 2-naphthols were determined in urine samples and used as biomarkers of JP-8 exposure. Multiple linear regression analyses were conducted to investigate the relative contributions of dermal and inhalation exposure to JP-8, and demographic and work-related covariates, to the levels of urinary naphthols. Our results show that both inhalation exposure and smoking significantly contributed to urinary 1-naphthol levels. The contribution of dermal exposure was significantly associated with levels of urinary 2-naphthol but not with urinary 1-naphthol among fuel-cell maintenance workers who wore supplied-air respirators. We conclude that dermal exposure to JP-8 significantly contributes to the systemic dose and affects the levels of urinary naphthalene metabolites. Future work on dermal xenobiotic metabolism and toxicokinetic studies are warranted in order to gain additional knowledge on naphthalene metabolism in the skin and the contribution to systemic exposure.

Selecting plants and nitrogen rates to vegetate crude-oil-contaminated soil.

Phytoremediation can be effective for remediating contaminated soils in situ and generally requires the addition of nitrogen (N) to increase plant growth. Our research objectives were to evaluate seedling emergence and survival of plant species and to determine the effects of N additions on plant growth in crude-oil-contaminated soil. From a preliminary survival study, three warm-season grasses--pearlmillet (Pennisetum glaucum [L.] R. Br.), sudangrass (Sorghum sudanense [Piper] Stapf [Piper]), and browntop millet (Brachiaria ramosa L.)--and one warm-season legume--jointvetch (Aeschynomene americana L.)--were chosen to determine the influence of the N application rate on plant growth in soil contaminated with weathered crude oil. Nitrogen was added based on total petroleum hydrocarbon-C:added N ratios (TPH-C:TN) ranging from 44:1 to 11:1. Plant species were grown for 7 wk. Root and shoot biomass were determined and root length and surface area were analyzed. Pearlmillet and sudangrass had higher shoot and root biomass when grown at a TPH-C:TN (inorganic) ratio of 11:1 and pearlmillet had higher root length and surface area when grown at 11:1 compared with the other species. By selecting appropriate plant species and determining optimum N application rates, increased plant root growth and an extended rhizosphere influence should lead to enhanced phytoremediation of crude-oil-contaminated soil.

Biocatalytic desulfurization of diesel oil in an air-lift reactor with immobilized Gordonia nitida CYKS1 cells.

A new type of air-lift reactor with immobilized Gordonia nitida CYKS1 cells on a fibrous support was designed and used for the biocatalytic desulfurization (BDS) of diesel oil. Its performance was evaluated at different phase ratios of the oil to the aqueous medium (or oil phase fractions) and different sucrose concentrations. When the reaction mixture contained 10% diesel oil (v/v), 61-67% of sulfur was removed as the sulfur content decreased from 202-250 to 76-90 mg L(-1) in 72 h. The sulfur content did not decrease any further because the remaining sulfur compounds were recalcitrant to BDS. During the desulfurization, the strain CYKS1 consumed hydrocarbons in the diesel oil, mainly n-alkanes with 10-26 carbons, as carbon source even though an easily available carbon source, sucrose, was supplied.

The metabolism of nonane, a JP-8 jet fuel component, by human liver microsomes, P450 isoforms and alcohol dehydrogenase and inhibition of human P450 isoforms by JP-8.

Nonane, a component of jet-propulsion fuel 8 (JP-8), is metabolized to 2-nonanol and 2-nonanone by pooled human liver microsomes (pHLM). Cytochrome P450 (CYP) isoforms 1A2, 2B6 and 2E1 metabolize nonane to 2-nonanol, whereas alcohol dehydrogenase, CYPs 2B6 and 2E1 metabolize 2-nonanol to 2-nonanone. Nonane and 2-nonanol showed no significant effect on the metabolism of testosterone, estradiol or N,N-diethyl-m-toluamide (DEET), but did inhibit carbaryl metabolism. JP-8 showed modest inhibition of testosterone, estradiol and carbaryl metabolism, but had a more significant effect on the metabolism of DEET. JP-8 was shown to inhibit CYPs 1A2 and 2B6 mediated metabolism of DEET, suggesting that at least some of the components of JP-8 might be metabolized by CYPs 1A2and/or 2B6.