Biomarkers limitations

Due to a nascent understanding of the use and interpretation of biomarkers, implementation of biomarkers as tools of exposure in the general population is very limited. A biomarker of exposure is a xenobiotic substance or its metabolite(s), or the product of an interaction between a xenobiotic agent and some target molecule(s) or cell(s) that is measured within a compartment of an organism (NAS/NRC 1989). The preferred biomarkers of exposure are generally the substance itself or substance-specific metabolites in readily obtainable body fluid(s) or excreta. However, several factors can confound the use and... 

A biomarker of susceptibility is an indicator of an inherent or acquired limitation of an organism s ability to respond to the challenge of exposure to a specific xenobiotic substance. It can be an intrinsic genetic or other characteristic or a preexisting disease that results in an increase in absorbed dose, a decrease in the biologically effective dose, or a target tissue response. If biomarkers of susceptibility exist, they are discussed in Section 3.10 Populations That Are Unusually Susceptible. 

The purpose of this chapter is to describe the analytical methods that are available for detecting, measuring, and/or monitoring methyl parathion, its metabolites, and other biomarkers of exposure and effect to methyl parathion. The intent is not to provide an exhaustive list of analytical methods. Rather, the intention is to identify well-established methods that are used as the standard methods of analysis. Many of the analytical methods used for environmental samples are the methods approved by federal agencies and organizations such as EPA and the National Institute for Occupational Safety and Health (NIOSH). Other methods presented in this chapter are those that are approved by groups such as the Association of Official Analytical Chemists (AOAC) and the American Public Health Association (APHA). Additionally, analytical methods are included that modify previously used methods to obtain lower detection limits and/or to improve accuracy and precision. 

The advantages of combining toxicity testing with chemical analysis when dealing with complex mixtures of environmental chemicals are clearly evident. More useful information can be obtained than would be possible if one or the other were to be used alone. However, chemical analysis can be very expensive, which places a limitation on the extent to which it can be used. There has been a growing interest in the development of new, cost-effective biomarker assays for assessing the toxicity of mixtures. Of particular interest are bioassays that incorporate mechanistic... 

Quinones, lipid-soluble substances involved in electron transport, can also be used as biomarkers. Lipski et al. (155) u.sed quinone analyses, physiological tests, and fatty acid profiles to differentiate Gram-negative non-fermentative bacteria isolated from biofilters. Quinone type was found to be an efficient method to group isolates prior to the analysis of results from the physiological tests. The detection of quinones appears to be restricted to the discrimination of isolated colonies and has limited potential to the analysis of mixed populations. 

A number of biochemical markers not associated with the cell envelope allow the specific detection of individual microorganisms in environmental samples. These include secondary alcohols. For example, Mycobacterium xenopi can be detected through the hydrolysis of wax ester mycolates, which liberates 2-docosanol, a characteristic and dominant secondary alcohol, which can be detected at low levels by GC-MS. This biomarker was found to be very useful for the rapid detection of M. xenopi in drinking water (159,160). Results from the GC-MS detection of 2-docosanol were obtained within 2 days compared to the 12 weeks required for culturable detection of M. xenopi. The detection limit for this type of approach was found to be 10 colony-forming units (CFU) ml" drinking water. 

Effect. No biomarkers have been identified to characterize the effects associated with exposure to diisopropyl methylphosphonate. Toxicity data for diisopropyl methylphosphonate are limited. No specific clinical signs or symptoms in humans have been positively linked to diisopropyl methylphosphonate exposure. Further research is needed to identify the mechanism of neurological effects and the clinical symptoms of diisopropyl methylphosphonate exposure. 

Mineral Oil Hydraulic Fluid. Limited studies were located that suggest biomarkers of exposure to mineral oil hydraulic fluids. No data that indicate quantitative or qualitative biomarkers of exposure to mineral oil hydraulic fluid were located. Mineral oil (hydrocarbons containing 15-30 carbon atoms per molecule) is a major component that is common to all mineral oil hydraulic fluids. Following exposure to food-grade mineral oil, most of the administered radioactivity was excreted in the feces as mineral oil (Ebert et al. 1966). Although the presence of mineral oil is a biomarker of exposure to mineral oil hydraulic fluids, it is also a biomarker of exposure to other readily available products that contain mineral oils. 

Mineral Oil Hydraulic Fluids and Polyalphaolefin Hydraulic Fluids. The data bases regarding the toxicity of mineral oil and polyalphaolefin hydraulic fluids are very limited. Further toxicity testing may elucidate biomarkers of exposure for these classes of hydraulic fluids. 

No data are available on platelet NTE activity in exposed subjects, and little data on lymphocyte NTE activity. In one reported case of suicidal poisoning with chlorpyrifos, inhibition of lymphocyte NTE was correlated with the enzyme inhibition in peripheral nerves (Osterloh et al., 1983). In another case of attempted suicide with the same compound, inhibition of NTE in peripheral lymphocytes was associated with the development of delayed neuropathy (Lotti, 1986). However, the threshold of NTE inhibition required for delayed neuropathy remains undetermined (Lotti, 1987). Observations in occupationally exposed subjects are limited in number, and more research is needed to investigate the applicability of NTE as a biomarker of exposure to OP pesticides. 

The lack of specificity of this biomarker and the complexity of analytical procedures for CS2 determination represent major limitations to the practical use of this metabolite to monitor occupational exposure. Moreover, only few data are available to confirm the validity of CS2 as a biomarker of DTC exposure. 

Encouraged by this spectral reproducibility, we focused our efforts on the particularly challenging problem of distinguishing bacterial strains by MALDI MS. We developed a modified correlation approach22 that relies on two fundamental qualities of bacterial mass spectra. First, because different bacterial strains of the same species have substantial, if not complete, genetic overlap, most of the protein masses observed with two different strains will be identical. This feature limits the value of the biomarker approach that is commonly used to differentiate bacteria species. Second, as just noted, closely controlled sample preparation and mass analysis procedures can result in highly reproducible results.22 The modified correlation approach takes advantage of subtle, yet reproducible, differences in mass spectra obtained from dif-... 

Flaag et al. found that several species of Haemophilus bacteria can be distinguished by MALDI-TOF mass spectrometry and further showed that different strains of Haemophilus ducreyi tend to exhibit specific unique biomarker peaks between 5000 and 15000 Da.60 Lynn et al. observed a few peaks that could distinguish E. coli strains 0 157 and 7G1, though they specifically note that attempts to identify bacteria based on a limited number of biomarkers can lead to errors.61... 

Intact bacteria were first introduced into a mass spectrometer for analysis of molecular biomarkers without processing and fractionation around 1975.6 The ionization techniques available at the time limited analysis to secondary metabolites that could be volatilized, such as quinines and diglycerides, and vigorous pyrolysis of bacteria was explored as an alternative.7 Although biomarkers were destroyed in pyrolysis strategies, computer-supported cluster analysis was developed to characterize pure samples. 

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