Biomedical subjects

Peter Adriaens

Publications and source records attributed to Peter Adriaens.

10 recordsLinked to original sources

PubMed2005

Comparing recovering efficiency of immunomagnetic separation and centrifugation of mycobacteria in metalworking fluids.

The accurate detection and enumeration of Mycobacterium immunogenum in metalworking fluids (MWFs) is imperative from an occupational health and industrial fluids management perspective. We report here a comparison of immunomagnetic separation (IMS) coupled to flow-cytometric enumeration, with traditional centrifugation techniques for mycobacteria in a semisynthetic MWF. This immunolabeling involves the coating of laboratory-synthesized nanometer-scale magnetic particles with protein A, to conjugate a primary antibody (Ab), specific to Mycobacterium spp. By using magnetic separation and flow-cytometric quantification, this approach enabled much higher recovery efficiency and fluorescent light intensities in comparison to the widely applied centrifugation technique. This IMS technique increased the cell recovery efficiency by one order of magnitude, and improved the fluorescence intensity of the secondary Ab conjugate by 2-fold, as compared with traditional techniques. By employing nanometer-scale magnetic particles, IMS was found to be compatible with flow cytometry (FCM), thereby increasing cell detection and enumeration speed by up to two orders of magnitude over microscopic techniques. Moreover, the use of primary Ab conjugated magnetic nanoparticles showed better correlation between epifluorescent microscopy counts and FCM analysis than that achieved using traditional centrifugation techniques. The results strongly support the applicability of the flow-cytometric IMS for microbial detection in complex matrices.

Centrifugation↗

PubMed2005

Microbial dechlorination of dioxins in estuarine enrichment cultures: effects of respiratory conditions and priming compound on community structure and dechlorination patterns.

The effect of respiratory conditions and priming compound on dechlorination patterns of heptachlorodibenzo-p-dioxins (HpCDD) was investigated using estuarine sediment-eluted cultures in the presence and absence of 20 mM sulfate, and 0.2 microM 2-bromodibenzo-p-dioxin (2-BrDD) as a priming compound. Electron balance calculations based on fatty acid turnover, hydrogen production, and electron acceptor depletion/methane formation indicated that whereas fermentative processes dominated in sulfate-free incubations, sulfate-reduction was predominant in the sulfate-amended incubations. The dechlorination of 1,2,3,4,6,7,8-HpCDD exhibited the following trends: (i) the relative yields of product formation did not exceed 30% and the presence of 2-BrDD increased the yield by up to 10%; (ii) sulfidogenic conditions resulted in a lower 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) formation, and the presence of 2-BrDD decreased the formation of 2,3,7,8-TCDD by additional 4-5-fold; (iii) the presence of 2-BrDD effected a predominance in lateral (2,3,7,8 positions) over peri (1,4,6,9 positions)-dechlorination. Denaturing gradient gel electrophoresis (DGGE) banding patterns indicated significant shifts of microbial community structure in response to terminal electron accepting processes as well as to the presence of the priming compound. The latter resulted in a similar community structure independent of dioxin spike, indicating that subsets of populations in the sediment are capable of exploiting the new niche provided by the priming compound.

Anaerobiosis↗

PubMed2004

Modified polytopic vector analysis to identify and quantify a dioxin dechlorination signature in sediments. 1. Theory.

Risk-based sediment management decisions require the characterization of contamination sources and fate processes in the field. Polytopic vector analysis (PVA) is a multivariate technique based on a linear mixing model, used to resolve chemical fingerprints and suited for forensic investigations of environmental contamination. The traditional algorithm is constrained to positive fingerprint (end-member) components and cannot resolve fingerprints with both positive and negative values required for a reactive end-member. We developed a modified algorithm (M-PVA) to resolve a dioxin dechlorination fingerprint, indicative of biotic/abiotic transformations in field samples of sediments. The new procedure isolates from the dioxin pattern net compositional changes due to dechlorination in a separate end-member. Using two artificial data sets for which the composition and sample contribution of all end-members are known, the dechlorination fingerprint was reproduced with a root mean square error of 28-41%. The dechlorination end-member contribution to the total variability (set at 4.0 and 10.0%, respectively) was overestimated 1-5-fold. The ability of M-PVA to reproduce the dechlorination pattern and its variability contribution depends on the actual contribution of dechlorination to variability. At an actual contribution of 4.0%, the model outcome deviates more strongly from the original than is the case for a contribution of 10.0%. As such, application of M-PVA to environmental data should include an uncertainty analysis to distinguish variability due to dechlorination from variability due to error. The development of the modified PVA procedure is an important step toward the field characterization of fate processes in dioxin-impacted sediments.

Algorithms↗

PubMed2004

Modified polytopic vector analysis to identify and quantify a dioxin dechlorination signature in sediments. 2. Application to the Passaic River.

Persistent contaminants such as dioxins have been documented to undergo dechlorination reactions in the laboratory; however, little is known about the importance of these reactions in the field. Polytopic vector analysis (PVA) is a statistical pattern recognition technique for multivariate data traditionally used to identify fingerprints of contaminant sources. A modified PVA algorithm with uncertainty analysis was used to model dechlorination fingerprints and sources. The technique was applied to 351 sediment core-derived dioxin samples from the lower reach of the Passaic River, New Jersey. A dechlorination fingerprint was identified with a highly positive 2,3,7,8-tetraCDD component and a highly negative heptaCDD component. The most important industrial source of 2,3,7,8-tetraCDD is a fingerprint related to 2,4,5-trichlorophenoxyacetic acid production. The dechlorination contribution to the data variance is 3.00 +/- 1.00%, corresponding to an average of 1.2 microg/kg of 2,3,7,8-tetraCDD per sample at the expense of heptaCDD. The possible occurrence of dechlorination was validated by comparing the local dechlorination contribution in the results to the value of the ratio 2,3,7,8-tetraCDD/total 2,3,7,8-PCDD, which indicates dechlorination in the laboratory. Bootstrap uncertainty analysis yielded the same dechlorination EM in 40% of the realizations. The results indicated that bootstrapping is an important statistical tool to quantify uncertainties with respect to the dechlorination EM and some of the source EMs.

Algorithms↗

PubMed2004

Carbon tetrachloride transformation on the surface of nanoscale biogenic magnetite particles.

Iron-reducing conditions in subsurface environments promote dechlorination reactions via both biotic and abiotic pathways, the latter often mediated via biologically activated minerals formed by dissimilatory iron-reducing bacteria (DIRB). Here we report the major products and pathways associated with the abiotic transformation of carbon tetrachloride (CT) by nanoscale biogenic magnetite/maghemite particles produced by the DIRB Geobacter metallireducens. Product formation and free radical/carbene trapping studies indicate that CT transformation occurs via three parallel pathways. The first pathway (hydrogenolysis) results in the formation of chloroform (45-50%) via a trichloromethyl free radical (*CCl3) and possibly a trichloromethyl carbanion (**CCl3-). The second and third pathways involve a dichlorocarbene intermediate (**CCl2), which either hydrolyzes to form CO (approximately 38%) (carbene hydrolysis), or undergoes further reduction to yield methane (8-10%) (carbene reduction). The mechanism of methane formation from **CCl2 is not known, but is speculated to involve a sequence of surface coordinated carbenoid and free radical complexes. The large fraction of relatively benign products formed by the carbene-mediated pathways suggests that magnetite/maghemite particles may have a beneficial application in the remediation of CT contaminated environments.

Carbon Tetrachloride↗

PubMed2003

Quantitative structure-activity relationship (QSAR) analysis of aromatic effector specificity in NtrC-like transcriptional activators from aromatic oxidizing bacteria.

A quantitative structure-activity relationship (QSAR) approach was taken to provide mechanistic insights into the interaction between the chemical structure of inducing compounds and the transcriptional activation of aromatic monooxygenase operons among the XylR/DmpR subclass of bacterial NtrC-like transcriptional regulators. Compared to XylR and DmpR, a broader spectrum of effector compounds was observed for the TbuT system from Ralstonia pickettii PKO1. The results of QSAR analysis for TbuT suggested that a steric effect, rather than hydrophobic or electronic effects, may be the predominant factor in determining aromatic effector specificity, and the active site of the regulator may positively interact not only with the methyl moiety but also with the most electron-rich aryl side of an aromatic effector.

Bacterial Proteins↗

PubMed2003

Intelligent infrastructure for sustainable potable water: a roundtable for emerging transnational research and technology development needs.

PROBLEM STATEMENT: Recent commercial and residential development have substantially impacted the fluxes and quality of water that recharge the aquifers and discharges to streams, lakes and wetlands and, ultimately, is recycled for potable use. Whereas the contaminant sources may be varied in scope and composition, these issues of urban water sustainability are of public health concern at all levels of economic development worldwide, and require cheap and innovative environmental sensing capabilities and interactive monitoring networks, as well as tailored distributed water treatment technologies. To address this need, a roundtable was organized to explore the potential role of advances in biotechnology and bioengineering to aid in developing causative relationships between spatial and temporal changes in urbanization patterns and groundwater and surface water quality parameters, and to address aspects of socioeconomic constraints in implementing sustainable exploitation of water resources. WORKSHOP OUTCOMES: An interactive framework for quantitative analysis of the coupling between human and natural systems requires integrating information derived from online and offline point measurements with Geographic Information Systems (GIS)-based remote sensing imagery analysis, groundwater-surface water hydrologic fluxes and water quality data to assess the vulnerability of potable water supplies. Spatially referenced data to inform uncertainty-based dynamic models can be used to rank watershed-specific stressors and receptors to guide researchers and policymakers in the development of targeted sensing and monitoring technologies, as well as tailored control measures for risk mitigation of potable water from microbial and chemical environmental contamination. The enabling technologies encompass: (i) distributed sensing approaches for microbial and chemical contamination (e.g. pathogens, endocrine disruptors); (ii) distributed application-specific, and infrastructure-adaptive water treatment systems; (iii) geostatistical integration of monitoring data and GIS layers; and (iv) systems analysis of microbial and chemical proliferation in distribution systems. IMPACT: This operational framework is aimed at technology implementation while maximizing economic and public health benefits. The outcomes of the roundtable will further research agendas in information technology-based monitoring infrastructure development, integration of processes and spatial analysis, as well as in new educational and training platforms for students, practitioners and regulators. The potential for technology diffusion to emerging economies with limited financial resources is substantial.

Conservation of Natural Resources↗

PubMed2003

Activity assessment of microorganisms eluted from sediments using 5-cyano-2,3-ditolyl tetrazolium chloride: a quantitative comparison of flow cytometry to epifluorescent microscopy.

Enhanced natural recovery may be successfully implemented at contaminated sediment sites, which are often characterized by large volumes of sediments with low to moderate levels of contamination to cost-effectively reduce human and ecological risks. In order to evaluate the potential for microbial contribution to remediation strategies, physiological assessment of indigenous microorganisms is essential. We report here a method for rapid and accurate assessment of metabolically (5-cyano-2,3-ditolyl tetrazolium chloride [CTC]) active microorganisms eluted from sediment, based on flow cytometry (FCM). Microorganisms eluted from sediment and suspended in estuarine medium were stained with CTC and counterstained with the DNA stain Picogreen (PG). Optimal stain concentrations and incubation times were employed. FCM quantification of the dual-stained microorganisms was not statistically different (paired t test; alpha=0.05; df=10) from enumeration (total or active numbers) by an established method (fluorescent microscopy) over two orders of magnitude (approximately 10(4)-10(6)/ml). This research suggests that FCM, which allows the collection and analysis of multiple parameters (light scatter and fluorescence emission), is a good candidate for microbial characterization in complex environmental matrices, such as sediments, across a broad range of activity levels (approximately 2% to 84% of total). Potential applications for this FCM-based method include the rapid assessment of changes in sediment microbial activity in response to enhanced bioremediation strategies.

DNA, Bacterial↗

PubMed2003

Quantitative structure-biodegradation relationships for ortho-substituted biphenyl compounds oxidized by Methylosinus trichosporium OB3b.

Methanotrophs, bacteria that thrive in the presence of stable methane and oxygen concentrations, can cometabolically oxidize ortho-substituted biphenyls to yield a variety of hydroxylated products. Despite awareness of the susceptibility of ortho-substituted biphenyls and other aromatic compounds to methanotrophic oxidation, the molecular properties relevant for predicting rates of methanotrophic oxidation are unknown. To this end, we have developed quantitative structure-biodegradation relationships using oxygen uptake activity by the type 2 methanotroph. Methylosinus trichosporium OB3b, expressing the soluble form of methane monooxygenase and in the presence of nine ortho-substituted biphenyls. Multivariate analysis yielded the strongest correlations using the initial slope of the oxygen uptake rate versus substrate concentration curve as the dependent variable. Quantum mechanical descriptors, including the sum of carbon charges on the substituted ring, the charge on the substituted carbon, and the width of compound calculated using computationally derived bond lengths and dihedral angles, correlated more strongly with oxygen uptake activity than did empirically derived electronic descriptors. The resulting models suggest a significant influence of substituent electronic nature and size and the involvement of the substituted carbon site in the oxidation of these compounds by M. trichosporium OB3b.

Biodegradation, Environmental↗

PubMed2002

Carbon tetrachloride transformation in a model iron-reducing culture: relative kinetics of biotic and abiotic reactions.

Contributions of biotic (cell-mediated) and abiotic (mineral-mediated) reactions to carbon tetrachloride (CT) transformation were studied in a model iron-reducing system that used hydrous ferric oxide (HFO) as the electron acceptor, acetate as the substrate, and Geobacter metallireducens as a representative dissimilative iron-reducing bacteria (DIRB). Over a period of 2-3 weeks, nanoscale magnetite particles, Fe3O4, were consistently formed as a product of iron respiration in this system. CT transformation rates were measured independently in resting cell suspensions of G. metallireducens or in suspensions of washed magnetite particles recovered from spent cultures. Protein and surface area-normalized expressions were derived for the biotic and abiotic reaction rates, respectively. Using the yield of total protein and magnetite surface area formed during growth in the model system as a basis for comparison, the mineral-mediated (abiotic) reaction was estimated to be 60-260-fold faster than the biotic reaction throughout the incubation period. We conclude that G. metallireducens induces CT transformation in this system primarily through the formation of reactive mineral surfaces rather than via co-metabolic mechanisms. The findings indicate that reactive biogenic minerals could play a significant role in the natural attenuation of chlorinated solvents in iron-reducing environments. A novel approach for stimulating reductive transformation of contaminants may be to enhance the formation of reactive biogenic minerals in situ.

Biotransformation↗