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Biomedical subjects

Peter G Green

Publications and source records attributed to Peter G Green.

12 recordsLinked to original sources

Using an integrated approach to link biomarker responses and physiological stress to growth impairment of cadmium-exposed larval topsmelt.

In this study, we used an integrated approach to determine whether key biochemical, cellular, and physiological responses were related to growth impairment of cadmium (Cd)-exposed larval topsmelt (Atherinops affinis). Food intake (Artemia franciscana nauplii), oxygen consumption rates, apoptotic DNA fragmentation (TUNEL assay), and metallothionein (MT)-like protein levels, were separately measured in relation to growth of larval topsmelt aqueously exposed to sublethal doses of Cd for 14 days. Cadmium accumulation and concentrations of abundant metals were also evaluated in a subset of fish. Fish in the highest Cd treatments (50 and 100 ppb Cd) were smaller in final mean weight and length, and consumed fewer A. franciscana nauplii than control fish. Food intake was positively correlated with final weight of larval topsmelt in Cd and control treatments; food intake increased as final weight of the fish increased. Oxygen consumption rates were positively correlated with Cd concentration and mean oxygen consumption rates were inversely correlated with final mean weight of topsmelt; the smallest fish were found in the highest Cd treatment and were respiring at higher rates than control fish. Apoptotic DNA fragmentation was concentration-dependent and was associated with diminished growth. Apoptotic DNA fragmentation was elevated in the gill of fish exposed to 50 ppb Cd, and in the gut, gill, and liver of fish exposed to 100 ppb Cd. Metallothionein (MT)-like protein levels in fish from 100 ppb Cd treatments were significantly higher than those in other treatments. Oxygen consumption rates may have increased as a compensatory response to Cd exposure. However, it is likely that the energy produced was allocated to an increased metabolic demand due to apoptosis, MT synthesis, and changes in ion regulation. This diversion of energy expenditures could contribute to growth impairment of Cd-exposed fish.

Animals↗

LC-MS analysis of carbonyl compounds and their occurrence in diesel emissions.

Liquid chromatography coupled with atmospheric pressure chemical ionization (APCI) ion trap mass spectrometry (ITMS) is applied to atmospheric aerosol relevant carbonyls. Characterization of positive and negative ion detection mass spectra are presented for 24 model compounds analyzed in their underivatized and O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) oxime forms. The addition of PFBHA derivatization enhanced the detection and sensitivity for many of the carbonyls investigated. For all but five of the carbonyls examined, a pseudomolecular (M + H)+ ion is the base peak in the APCI positive ion mass spectra of PFBHA oxime derivatives and is observed in four of the five exceptions. Application of the evaluated analysis methodology to heavy-duty diesel source emissions facilitated the quantification of 10 aliphatic carbonyls (5 C5-C9 ketones, 4 C6 unsaturated ketones, 1 C6 dicarbonyl) and 14 aromatic carbonyls (1 C9 aldehyde, 5 C8-C13 ketones, 8 C6-C14 quinones). Diesel truck engine emission factors spanning 0.55-540 microg km(-1) were measured for gas- and particle-phase carbonyls. Good agreement was observed for gas-phase emission factors with results obtained by gas chromatography with ITMS.

Chromatography, Liquid↗

Seasonal and spatial patterns of metals at a restored copper mine site. I. Stream copper and zinc.

Seasonal and spatial variations in metal concentrations and pH were found in a stream at a restored copper mine site located near a massive sulfide deposit in the Foothill copper-zinc belt of the Sierra Nevada, California. At the mouth of the stream, copper concentrations increased and pH decreased with increased streamflow after the onset of winter rain and, unexpectedly, reached extreme values 1 or 2 months after peaks in the seasonal hydrographs. In contrast, aqueous zinc and sulfate concentrations were highest during low-flow periods. Spatial variation was assessed in 400 m of reach encompassing an acidic, metal-laden seep. At this seep, pH remained low (2-3) throughout the year, and copper concentrations were highest. In contrast, the zinc concentrations increased with downstream distance. These spatial patterns were caused by immobilization of copper by hydrous ferric oxides in benthic sediments, coupled with increasing downstream supply of zinc from groundwater seepage.

California↗

Seasonal and spatial patterns of metals at a restored copper mine site II. Copper in riparian soils and Bromus carinatus shoots.

Soil and plants were sampled throughout winter and spring near a perennial stream traversing a restored mine site in a winter-rainy climate. Within 1m of an acidic reach of the stream, soil had pH 3-5 and 50-100 microg/g "bioavailable" copper (extractable with 0.01 M CaCl2). Soil 2-3 m from the stream had pH 5-8 and lower (less than 3 microg/g) bioavailable copper. "Oxide-bound" copper (extractable with 2N HCl) was 50-100 microg/g at most locations. Copper concentrations in the shoots of field-collected Bromus carinatus declined from 20 microg/g in winter to 2 microg/g in spring at all sampling sites. A similar temporal pattern was found in plants grown under controlled conditions. Thus B. carinatus has a developmental program for control of shoot copper concentration, causing a seasonally-varying pattern of copper phytoaccumulation over a large range of copper availability in the soil.

Biodegradation, Environmental↗

Measuring the trace elemental composition of size-resolved airborne particles.

A new method to measure the trace elemental composition of size-resolved airborne particles that uses acetone extraction followed by ICPMS analysis is compared to three other established methods: copper anode XRF, molybdenum anode XRF, and an ICPMS method that uses HF digestion. The method detection limit (MDL), accuracy, and precision of each method is studied through the analysis of ambient samples collected in California. The MDLs of the new acetone-ICPMS method are similar to MDLs for the established HF-ICPMS method. Both sets of ICPMS MDLs are 1-3 orders of magnitude lower than XRF MDLs for approximately 50 elements other than the light crustal elements such as silicon, sulfur, calcium, and zinc. The accuracy of the acetone-ICPMS method was verified by comparison to measurements made using ion chromatography and the HF-ICPMS method. The acetone-ICPMS analysis method was more precise than the conventional HF-ICPMS method for collocated measurements. Both ICPMS methods were more precise than XRF for most elements. The size distribution of 21 elements contained in ambient particles collected with cascade impactors could be measured with good precision using the new acetone-ICPMS analysis method: lithium, sulfur, potassium, titanium, vanadium, manganese, iron, gallium, germanium, arsenic, selenium, bromine, rubidium, strontium, cadmium, tin, antimony, barium, thallium, lead, and bismuth. It is likely that the size distribution of an additional 9 elements could also be measured when concentrations are sufficiently high: phosphorus, molybdenum, niobium, palladium, cesium, europium, holmium, platinum, and uranium. None of the conventional methods were able to measure the size distribution of these elements with acceptable precision under the conditions studied. The new acetone-ICPMS method should provide useful data for the study of the health effects of airborne particles.

Air Pollution↗

Tidal salt marsh sediment in California, USA. Part 2: occurrence and anthropogenic input of trace metals.

Surface sediment samples (0-5 cm) from 5 tidal salt marshes along the coast in California, USA were analyzed to investigate the occurrence and anthropogenic input of trace metals. Among study areas, Stege Marsh located in the central San Francisco Bay was the most contaminated marsh. Concentrations of metals in Stege Marsh sediments were higher than San Francisco Bay ambient levels. Zinc (55.3-744 microg g(-1)) was the most abundant trace metal and was followed by lead (26.6-273 microg g(-1)). Aluminum normalized enrichment factors revealed that lead was the most anthropogenically impacted metal in all marshes. Enrichment factors of lead in Stege Marsh ranged from 8 to 49 (median=16). Sediments from reference marshes also had high enrichment factors (2-8) for lead, indicating that lead contamination is ubiquitous, possibly due to continuous input from atmospherically transported lead that was previously used as a gasoline additive. Copper, silver, and zinc in Stege Marsh were also enriched by anthropogenic input. Though nickel concentrations in Stege Marsh and reference marshes exceeded sediment quality guidelines, enrichment factors indicated nickel from anthropogenic input was negligible. Presence of nickel-rich source rock such as serpentinite in the San Francisco Bay watershed can explain high levels of nickel in this area. Coefficients of variation were significantly different between anthropogenically impacted and non-impacted metals and might be used as a less conservative indicator for anthropogenic input of metals when enrichment factors are not available.

California↗

Tidal salt marsh sediment in California, USA. Part 1: occurrence and sources of organic contaminants.

Surface sediment samples (0-5 cm) from five tidal marshes along the coast of California, USA were analyzed for organic pollutants to investigate their relationship to land use, current distribution within marshes, and possible sources. Among the study areas, Stege Marsh, located in San Francisco Bay, was the most contaminated. Compared to San Francisco Bay, Stege Marsh had much higher levels of organic contaminants such as PCBs (polychlorinated biphenyls), DDTs, and chlordanes. At reference marshes (Tom's Point and Walker Creek in Tomales Bay), organic contaminants in sediments were very low. While PAHs (polycyclic aromatic hydrocarbons) were found at all of the study areas (22-13,600 ng g(-1)), measurable concentrations of PCBs were found only in the sediments from Stege Marsh (80-9,940 ng g(-1)). Combustion related (pyrogenic) high molecular weight PAHs were dominant in sediments from Stege and Carpinteria Marshes, while in sediments from Tom's Point and Walker Creek petroleum related (petrogenic) low molecular weight PAHs and alkyl-substituted PAHs were much more abundant than pyrogenic PAHs. PCB congener patterns in all of the Stege Marsh samples were the same and revealed that Aroclor 1248 was a predominant source. In all marshes, the sum of DDE and DDD accounted for more than 90% of total DDTs, indicating that DDT has degraded significantly. The ratios of p,p'-DDE to p,p'-DDD in sediments from Stege Marsh provide evidence of possible previous use of technical DDD. Chlordane ratios indicated that chlordanes have degraded slightly. Bis(2-ethylhexyl)phthalate (280-32,000 ng g(-1)) was the most abundant phthalate. The data indicates that Stege Marsh may be a source of contaminants that continue to be discharged into San Francisco Bay.

California↗

Mercury methylation from unexpected sources: molybdate-inhibited freshwater sediments and an iron-reducing bacterium.

Methylmercury has been thought to be produced predominantly by sulfate-reducing bacteria in anoxic sediments. Here we show that in circumneutral pH sediments (Clear Lake, CA) application of a specific inhibitor of sulfate-reducing bacteria at appropriate concentrations typically inhibited less than one-half of all anaerobic methylation of added divalent mercury. This suggests that one or more additional groups of microbes are active methylators in these sediments impacted by a nearby abandoned mercury mine. From Clear Lake sediments, we isolated the iron-reducing bacterium Geobacter sp. strain CLFeRB, which can methylate mercury at a rate comparable to Desulfobulbus propionicus strain 1pr3, a sulfate-reducing bacterium known to be an active methylator. This is the first time that an iron-reducing bacterium has been shown to methylate mercury at environmentally significant rates. We suggest that mercury methylation by iron-reducing bacteria represents a previously unidentified and potentially significant source of this environmental toxin in iron-rich freshwater sediments.

DNA, Ribosomal↗

Diversity, composition, and geographical distribution of microbial communities in California salt marsh sediments.

The Pacific Estuarine Ecosystem Indicators Research Consortium seeks to develop bioindicators of toxicant-induced stress and bioavailability for wetland biota. Within this framework, the effects of environmental and pollutant variables on microbial communities were studied at different spatial scales over a 2-year period. Six salt marshes along the California coastline were characterized using phospholipid fatty acid (PLFA) analysis and terminal restriction fragment length polymorphism (TRFLP) analysis. Additionally, 27 metals, six currently used pesticides, total polychlorinated biphenyls and polycyclic aromatic hydrocarbons, chlordanes, nonachlors, dichlorodiphenyldichloroethane, and dichlorodiphenyldichloroethylene were analyzed. Sampling was performed over large (between salt marshes), medium (stations within a marsh), and small (different channel depths) spatial scales. Regression and ordination analysis suggested that the spatial variation in microbial communities exceeded the variation attributable to pollutants. PLFA analysis and TRFLP canonical correspondence analysis (CCA) explained 74 and 43% of the variation, respectively, and both methods attributed 34% of the variation to tidal cycles, marsh, year, and latitude. After accounting for spatial variation using partial CCA, we found that metals had a greater effect on microbial community composition than organic pollutants had. Organic carbon and nitrogen contents were positively correlated with PLFA biomass, whereas total metal concentrations were positively correlated with biomass and diversity. Higher concentrations of heavy metals were negatively correlated with branched PLFAs and positively correlated with methyl- and cyclo-substituted PLFAs. The strong relationships observed between pollutant concentrations and some of the microbial indicators indicated the potential for using microbial community analyses in assessments of the ecosystem health of salt marshes.

Bacteria↗

Validation of otolith growth rate analysis using cadmium-exposed larval topsmelt (Atherinops affinis).

We applied otolith growth rate analysis to an investigation of cadmium (Cd)-exposed larval topsmelt (Atherinops affinis) to determine if growth rate was a more sensitive measure than somatic growth (body wt or length). Topsmelt otoliths, calcareous concretions in the fish inner ear, formed daily increments, and otolith growth was proportional to somatic growth. Nine-day posthatch larval topsmelt were exposed to Cd (0-100 ppb) in seawater for 14 d and fed low or high ration levels in separate experiments. Whereas Cd impaired topsmelt growth and growth rates, the extent of growth reduction was dependent on the ration level. At high ration levels, otolith and somatic growth rates of fish exposed to Cd (50 and 100 ppb) were significantly reduced; however, no differences in final mean weight and only marginal differences in final mean length of Cd-exposed topsmelt were observed. At low ration levels, we detected reductions in both somatic growth as well as otolith and somatic growth rates of topsmelt exposed to Cd (50 and 100 ppb). Otolith growth rate analysis was more sensitive than growth measurements of Cd-exposed topsmelt, because it allowed the detection of small differences in growth rates even when differences in somatic growth were not observed.

Animals↗

Passive sampling screening method using thin-layer chromatography plates.

An in situ solid-phase extraction method, thin-layer chromatography (TLC) passive sampling, was investigated as a screening method for determining the presence of organic compounds in water using laboratory experiments and field applications. The TLC passive sampler developed in this research enables the spatial and temporal distributions of organic compounds to be determined qualitatively with little expense. The materials for the sampler developed here cost dollar 1.60 each. By first identifying the areas where a pollutant is present using the TLC passive sampling screening method, total analytical costs for monitoring programs may be reduced by eliminating unnecessary conventional analyses for locations where the pollutant is not present. Two organophosphorus pesticides, diazinon and chlorpyrifos, were used as a model for the development and as a potential application of the TLC passive sampling method. Passive sampler adsorption kinetics, enrichment factors, and extraction efficiencies were determined from batch experiments with exposure times ranging from 1 h to 4 weeks. In field applications, TLC passive sampling was conducted in natural and engineered systems with two silica gel extraction media, C2 and C18. Diazinon and chlorpyrifos were detected by analyzing the adsorbed compounds by high-performance liquid chromatography with ultraviolet detection.

Chromatography, High Pressure Liquid↗

Biological activity of inorganic arsenic and antimony reflects oxidation state in cultured human keratinocytes.

Sodium arsenite is much more potent than sodium arsenate in producing adverse effects in animals and in cultured cells. Although arsenate may exhibit toxicity as a phosphate analogue, its potency in vivo appears to be enhanced by reduction to arsenite. To understand the relative importance of this reduction, which is critical in evaluating the responsiveness of cell culture models to the different oxidation states and thus to elucidating the mechanism of arsenic action, present work has correlated the extent of reduction with biological activity in human keratinocytes. The results show that at biologically relevant concentrations, arsenate reduction to appreciable levels required several days, helping rationalize a previous empirical observation that it was approximately one-third as potent as arsenite. The relatively low conversion rate also emphasizes a limitation of culture; arsenate was nearly as efficacious as arsenite, but the time required for it to reach maximal effect exceeded ordinary medium change intervals. In keratinocytes, an important role for purine nucleoside phosphorylase in the reduction could not be demonstrated, indicating that another pathway is dominant in this cell type. Methylation of inorganic arsenic, uptake of methylated forms, and their reduction were all very slow. These findings suggest that the reduced methylated forms have only a small contribution to skin carcinogenesis unless they are supplied through the circulation. In parallel experiments, trivalent antimony was similar to arsenite in potency and efficacy, whereas pentavalent antimony was virtually without biological effect. Conversion of antimony in the pentavalent to the trivalent oxidation state was not detectable in keratinocytes. These findings emphasize the importance of intracellular reduction of the metalloids for biological effects.

Animals↗