Biomedical subjects

P Lee Ferguson

Publications and source records attributed to P Lee Ferguson.

9 recordsLinked to original sources

PubMed2006

Life-cycle effects of single-walled carbon nanotubes (SWNTs) on an estuarine meiobenthic copepod.

Single-walled carbon nanotubes (SWNT) are finding increasing use in consumer electronics and structural composites. These nanomaterials and their manufacturing byproducts may eventually reach estuarine systems through wastewater discharge. The acute and chronic toxicity of SWNTs were evaluated using full life-cycle bioassays with the estuarine copepod Amphiascus tenuiremis (ASTM method E-2317-04). A synchronous cohort of naupliar larvae was assayed by culturing individual larvae to adulthood in individual 96-well microplate wells amended with SWNTs in seawater. Copepods were exposed to "as prepared" (AP) SWNTs, electrophoretically purified SWNTs, or a fluorescent fraction of nanocarbon synthetic byproducts. Copepods ingesting purified SWNTs showed no significant effects on mortality, development, and reproduction across exposures (p < 0.05). In contrast, exposure to the more complex AP-SWNT mixture significantly increased life-cycle mortality, reduced fertilization rates, and reduced molting success in the highest exposure (10 mg x L(-1)) (p < 0.05). Exposure to small fluorescent nanocarbon byproducts caused significantly increased life-cycle mortality at 10 mg x L(-1) (p < 0.05). The fluorescent nanocarbon fraction also caused significant reduction in life-cycle molting success for all exposures (p < 0.05). These results suggest size-dependent toxicity of SWNT-based nanomaterials, with the smallest synthetic byproduct fractions causing increased mortality and delayed copepod development over the concentration ranges tested.

Animals↗

PubMed2005

Application of ion trap-MS with H/D exchange and QqTOF-MS in the identification of microbial degradates of trimethoprim in nitrifying activated sludge.

In this work, the identification of two microbial degradation products of the antimicrobial trimethoprim (290 Da) is described. The structural elucidation of the metabolites, which were produced by nitrifying activated sludge bacteria in a small-scale laboratory batch reactor, was accomplished by electrospray ionization-ion trap mass spectrometry conducting consecutive fragmentation steps (MS(n)) combined with H/D-exchange experiments. Although one metabolite corresponded to alpha-hydroxytrimethoprim (306 Da), oxidation of the aromatic ring within the diaminopyrimidine substructure was determined for the second degradate (324 Da). Accurate mass measurements of the two metabolites were provided by a hybrid quadrupole time-of-flight-mass spectrometer operated in MS/MS mode. With absolute mass errors of <5 mDa, it allowed us to confirm the proposed elemental composition for the protonated precursor ions as well as for a series of fragment ions that were previously identified by ion trap mass spectrometry. The study emphasized the potential of nitrifying activated sludge bacteria for breaking down an environmentally relevant pharmaceutical that is otherwise poorly degradable by a bacterial community encountered in conventional activated sludge.

Anti-Infective Agents, Urinary↗

PubMed2004

Proteomic analysis of arginine adducts on glyoxal-modified ribonuclease.

Accumulation of advanced glycation end-products (AGEs) on proteins is associated with the development of diabetic complications. Although the overall extent of modification of protein by AGEs is limited, localization of these modifications at a few critical sites might have a significant effect on protein structure and function. In the present study, we describe the sites of modification of RNase by glyoxal under physiological conditions. Arg39 and Arg85, which are closest to the active site of the enzyme, were identified as the primary sites of formation of the glyoxal-derived dihydroxyimidazolidine and hydroimidazolone adducts. Lower amounts of modification were detected at Arg10, while Arg33 appeared to be unmodified. We conclude that dihydroxyimidazolidine adducts are the primary products of modification of protein by glyoxal, that Arg39 and Arg85 are the primary sites of modification of RNase by glyoxal, and that modification of arginine residues during Maillard reactions of proteins is a highly selective process.

Amino Acids↗

PubMed2003

Biogeochemistry of nonylphenol ethoxylates in urban estuarine sediments.

We have examined the concentrations and distributions of nonylphenol ethoxylate (NPEO) surfactants and their primary neutral metabolites in two dated sediment cores collected in 1988 and 1996 from a depositional area proximal to a wastewater treatment plant within Jamaica Bay, NY. Total NPEO concentrations ranged from >50 microg/g near the surface (4-6 cm, deposited ca. 1990) to below detection limits (<0.1 microg/g) at 50 cm depth (deposited ca. 1940). The general decrease in NPEO concentrations with increasing depth in sediment reflected increased commercial use of these compounds over the last 50 yr. NPEO ethoxymer distributions in recent sediments were dominated by NP(0-3)EO, consistent with the increased relative input of these particular ethoxymers to the estuary following the upgrade of local biological sewage treatment processes to full activated sludge in the late 1970s. NPEO ethoxymer profiles in deeper sediments were characterized by relatively higher proportions of unmetabolized, highly ethoxylated NPEOs. Depth profiles of NP1EO and NP in the upper portion of the sediment core showed evidence for in situ diagenetic conversion of NP1EO to NP. However, comparison of NPEO concentrations in selected strata from the core collected in 1996 with those in matched strata from a core collected from the same location in 1988 provided no evidence for in situ degradation of total NPEOs during the elapsed 8 yr between collection dates.

Detergents↗

PubMed2003

Proteome analysis by mass spectrometry.

The coupling of high-performance mass spectrometry instrumentation with highly efficient chromatographic and electrophoretic separations has enabled rapid qualitative and quantitative analysis of thousands of proteins from minute samples of biological materials. Here, we review recent progress in the development and application of mass spectrometry-based techniques for the qualitative and quantitative analysis of global proteome samples derived from whole cells, tissues, or organisms. Techniques such as multidimensional peptide and protein separations coupled with mass spectrometry, accurate mass measurement of peptides from global proteome digests, and mass spectrometric characterization of intact proteins hold great promise for characterization of highly complex protein mixtures. Advances in chemical tagging and isotope labeling techniques have enabled quantitative analysis of proteomes, and highly specific isolation strategies have been developed aimed at selective analysis of posttranslationally modified proteins.

Chromatography, Affinity↗

PubMed2003

Proteome analyses using accurate mass and elution time peptide tags with capillary LC time-of-flight mass spectrometry.

We describe the application of capillary liquid chromatography (LC) time-of-flight (TOF) mass spectrometric instrumentation for the rapid characterization of microbial proteomes. Previously (Lipton et al., Proc. Natl. Acad. Sci. U.S.A. 2002, 99, 11049) the peptides from a series of growth conditions of Deinococcus radiodurans have been characterized using capillary LC MS/MS and accurate mass measurements which are captured as an accurate mass and time (AMT) tag database. Using this AMT tag database, detected peptides can be assigned using measurements obtained on a TOF due to the additional use of elution time data as a constraint. When peptide matches are obtained using AMT tags (i.e., using both constraints) unique matches of a mass spectral peak occurs 88% of the time. Not only are AMT tag matches unique in most cases, the coverage of the proteome is high; approximately 3500 unique peptide AMT tags are found on average per capillary LC run. From the results of the AMT tag database search, approximately 900 ORFs detected using LC-TOFMS, with approximately 500 ORFs covered by at least two AMT tags. These results indicate that AMT database searches with modest mass and elution time criteria can provide proteomic information for approximately one thousand proteins in a single run of <3 h. The advantage of this method over using MS/MS based techniques is the large number of identifications that occur in a single experiment as well as the basis for improved quantitation. For MS/MS experiments, the number of peptide identifications is severely restricted because of the time required to dissociate the peptides individually. These results demonstrate the utility of the AMT tag approach using capillary LC-TOF MS instruments, and also show that AMT tags developed using other instrumentation can be effectively utilized.

Amino Acid Sequence↗

PubMed2003

Degradation of nonylphenol ethoxylates in estuarine sediment under aerobic and anaerobic conditions.

Nonylphenol ethoxylate (NPEO) surfactants and their metabolites are ubiquitous contaminants of the aquatic environment. Despite considerable interest in the environmental fate of these compounds due to concerns over toxicity and estrogenic activity, the pathways of NPEO degradation in sediments have not previously been reported, in spite of the fact that sediment appears to be an important sink for these compounds in the environment. In the present work, we have examined the rates and pathways of NPEO degradation in batch sediment slurry experiments using radiolabeled NPEO mixtures. Results suggest that NPEOs are more persistent in sediments under anaerobic conditions than in the presence of oxygen. In addition, it was illustrated that NPEO degradation proceeds via separate pathways in oxic and anoxic sediment. Discernible metabolites were identified and an overall mass balance for NPEO degradation in oxic and anoxic sediment was achieved. In contrast with previous studies, no evidence was observed for net production of nonylphenol from NPEOs during aerobic or anaerobic degradation. The observed relative rates at which NPEO ethoxymers disappeared in the sediment slurry experiments were consistent with previous reports for these compounds in sediment and other environmental media, although the absolute rates measured were somewhat faster than those reported for field sediments.

Aerobiosis↗

PubMed2002

Polycyclic aromatic hydrocarbon bioaccumulation by meiobenthic copepods inhabiting a superfund site: techniques for micromass body burden and total lipid analysis.

Microtechniques for polycyclic aromatic hydrocarbon (PAH) body burden and total lipid analysis were developed and applied to determine the first lipid-normalized bioaccumulation factors for a hydrophobic organic toxicant in a meiobenthic organism (0.063-0.500 mm) living in field-contaminated sediments. The total lipid microtechnique combines the standard Bligh-Dyer extraction method with a colorimetric quantification method for analysis of samples containing I to 50 microg lipid. The microtechnique for body burden analysis quantifies PAHs from tissue samples containing as little as 10 pg PAH. Fluoranthene, benz[a]anthracene, and benzo[a]pyrene biota-sediment accumulation factors (BSAFs) were determined for the meiobenthic copepod Microarthridion littorale living in an estuarine U.S. Environmental Protection Agency Superfund site. Gravid female, nongravid female, and male BSAFs were 0.82, 0.54, and 0.36, respectively, for fluoranthene; 0.50, 0.44, and 0.40, respectively, for benz[a]anthracene; and 0.09, 0.12, and 0.15, respectively, for benzo[a]pyrene. Comparison of nonlipid-normalized bioaccumulation factors (BAFs) to BSAFs indicates that M. littorale bioaccumulated PAHs on a gram lipid basis. The BSAFs declined consistently with increasing PAH log K(ow) for all copepod sex and reproductive stages. Sex- and stage-specific comparisons of BSAFs suggest that differences in lipid content and quality may lead to differences in BSAF values depending on PAH molecular weight and/or hydrophobicity.

Animals↗

PubMedDate not supplied

Estrogenic responses of larval sunshine bass (Morone saxatilis x M. Chrysops) exposed to New York City sewage effluent.

To determine the estrogenicity of effluents from sewage treatment plants (STPs) to larval fish, 2-day-old sunshine bass were exposed to effluents from three STPs serving New York City (NYC), varying in size and treatment level. Estrogenic response was evaluated by measuring vitellogenin (VTG) and estrogen receptor (ER) expression in cytosolic fractions of whole body homogenates. Concentrations of the presumptive endocrine disruptors in the effluents were also measured. VTG and ER levels in sewage-exposed fish were 3-5 times that observed in controls. Combined concentrations of estradiol and estrone ranged from 5 to 13 ng/l and nonylphenol-ethoxylate metabolites (NPEOs: 4-nonylphenol, and 1-, 2-, and 3-nonylphenol-ethoxylates) ranged from 180 to 470 microg/l in chlorinated effluent. Results indicate that both ER and VTG can be used as biomarkers for endocrine disruption in larval fish, and that 4-day exposure to sewage effluent is sufficient to elicit significant expression of these markers in sunshine bass larvae. The extremely higher concentrations of NPEOs found in effluent relative to hormones (approximately 40,000-fold) indicates that surfactant metabolites may be contributing significantly to the estrogenic effects observed.

Animals↗