Mental Versus Physical Workload

The concept of mental workload (MWL) has become increasingly important since modern semi-automated and computerized technologies may impose severe requirements on human mental or information-processing capabilities within both manufacturing and administrative tasks. Thus, especially for the domains of job analysis, evaluation of job requirements and job design, the conceptualization of mental workload has become even more important than that of traditional physical workload.

Definitions of Mental Workload

There is no agreed-upon definition of mental workload. The main reason is that there are at least two theoretically well-based approaches and definitions: (1) MWL as viewed in terms of the task requirements as an independent, external variable with which the working subjects have to cope more or less efficiently, and (2) MWL as defined in terms of an interaction between task requirements and human capabilities or resources (Hancock and Chignell 1986; Welford 1986; Wieland-Eckelmann 1992).

Although arising from different contexts, both approaches offer necessary and well-founded contributions to different problems.

The requirements resources interaction approach was developed within the context of personality-environment fit/misfit theories which try to explain interindividually differing responses to identical physical and psychosocial conditions and requirements. Thus, this approach may explain individual differences in the patterns of subjective responses to loading requirements and conditions, for example, in terms of fatigue, monotony, affective aversion, burnout or diseases (Gopher and Donchin 1986; Hancock and Meshkati 1988).

The task requirements approach was developed within those parts of occupational psychology and ergonomics which are predominantly engaged in task design, especially in the design of new and untried future tasks, or so-called prospective task design. The background here is the stress-strain concept. Task requirements constitute the stress and the working subjects try to adapt to or to cope with the demands much as they would to other forms of stress (Hancock and Chignell 1986). This task requirements approach tries to answer the question of how to design tasks in advance in order to optimize their later impact on the—often still unknown—employees who will accomplish these future tasks.

There are at least a few common characteristics of both conceptualizations of MWL.

1. MWL mainly describes the input aspects of tasks, that is to say, the requirements and demands made by the tasks on the employees, which might be used in forecasting the task outcome.
2. The mental aspects of MWL are conceptualized in terms of information processing. Information processing includes cognitive as well as motivational/volitional and emotional aspects, since the persons always will evaluate the demands which they have to cope with and, thus, will self-regulate their effort for processing.
3. Information-processing integrates mental processes, representations (for example, knowledge, or mental models of a machine) and states (for example, states of consciousness, degrees of activation and, less formally, mood).
4. MWL is a multidimensional characteristic of task requirements, since every task varies in a couple of interrelated but nevertheless distinct dimensions which separately must be dealt with in task design.
5. MWL will have a multidimensional impact which at least will determine (a) behaviour, for example, the strategies and the resulting performance, (b) perceived, subjective short-term well-being with consequences for health in the long run, and (c) psycho-physiological processes, for example, alterations of blood pressure at work, which may become long-term effects of a positive kind (promoting, say, fitness improvement) or of a negative kind (involving impairments or ill-health).
6. From the point of view of task design, MWL should not be minimized—as would be necessary in the case of carcinogenic air pollution—but optimized. The reason is that demanding mental task requirements are inevitable for well-being, health promotion and qualification since they offer the necessary activating impulses, fitness prerequisites and learning/training options. Missing demands on the contrary may result in deactivation, loss of physical fitness, de-qualification and deterioration of so-called intrinsic (task content-dependent) motivation. Findings in this area led to the technique of health and personality promoting task design (Hacker 1986).
7. MWL therefore, in any case, must be dealt with in task analysis, task requirement evaluation as well as in corrective and prospective task design.

Theoretical Approaches: Requirement-Resources Approaches

From the person-environment fit point of view, MWL and its consequences may be roughly categorized—as is shown in figure 1—into underload, properly fitting load, and overload. This categorization results from the relationships between task requirements and mental capabilities or resources. Task requirements may exceed, fit with or fail to be satisfied by the resources. Both types of misfit may result from quantitative or qualitative modes of misfit and will have qualitatively differing, but in any case negative, consequences (see figure 1).

Figure 1. Types and consequences of requirements-resources relationships

Some theories attempt to define MWL starting from the resource or capacity side of the requirements, namely, resources relationships. These resource theories might be subdivided into resource volume and resource allocation theories (Wieland-Eckelmann 1992). The amount of available capacity may come from a single source (single resource theories) which determines processing. The availability of this resource varies with arousal (Kahneman 1973). Modern multiple resource theories suppose a set of relatively independent processing resources. Thus, performance will depend on the condition whether the same resource or different resources are required simultaneously and concurrently. Different resources are, for example, encoding, processing or responding resources (Gopher and Donchin 1986; Welford 1986). The most critical problem for these types of theories is the reliable identification of one or more well-defined capacities for qualitatively different processing operations.

Resource allocation theories suppose qualitatively changing processing as a function of varying strategies. Depending on the strategies, differing mental processes and representations may be applied for task accomplishment. Thus, not the volume of stable resources but flexible allocation strategies become the key point of interest. Again, however, essential questions—especially concerning the methods of diagnosis of the strategies—remain to be answered.

Assessment of MWL: using requirement-resource approaches

A strict measurement of MWL at present would be impossible since well-defined units of measurement are lacking. But, to be sure, the conceptualization and the instruments for an assessment should meet the general quality criteria of diagnostic approaches, which have objectivity, reliability, validity and usefulness. However, as of now, only a little is known about the overall quality of proposed techniques or instruments.

There are a sizeable number of reasons for the remaining difficulties with assessing MWL according to the requirement-resource approaches (O’Donnell and Eggemeier 1986). An attempt at MWL assessment has to cope with questions like the following: is the task self-intended, following self-set goals, or is it directed with reference to an externally defined order? Which type of capacities (conscious intellectual processing, application of tacit knowledge, etc.) are required, and are they called upon simultaneously or sequentially? Are there different strategies available and, if so, which ones? Which coping mechanisms of a working person might be required?

The most often discussed approaches try to assess MWL in terms of:

1. required effort (effort assessment) approaches applying—in some versions psychophysiologically validated—scaling procedures such as those offered by Bartenwerfer (1970) or Eilers, Nachreiner and Hänicke (1986), or
2. occupied or, vice versa, residual mental capacity (mental capacity assessment) approaches applying the traditional dual task techniques as, for example, discussed by O’Donnell and Eggemeier (1986).

Both approaches are heavily dependent on the assumptions of single resource theories and consequently have to struggle with the above-mentioned questions.

Effort assessment. Such effort assessment techniques as, for example, the scaling procedure applied to a perceived correlate of the general central activation, developed and validated by Bartenwerfer (1970), offer verbal scales which may be completed by graphic ones and which grade the unidimensionally varying part of the perceived required effort during task accomplishment. The subjects are requested to describe their perceived effort by means of one of the steps of the scale provided.

The quality criteria mentioned above are met by this technique. Its limitations include the unidimensionality of the scale, covering an essential but questionable part of perceived effort; the limited or absent possibility of forecasting perceived personal task outcomes, for example, in terms of fatigue, boredom or anxiety; and especially the highly abstract or formal character of effort which will identify and explain nearly nothing of the content-dependent aspects of MWL as, for example, any possible useful applications of the qualification or the learning options.

Mental capacity assessment. The mental capacity assessment consists of the dual task techniques and a related data interpretation procedure, called the performance operating characteristic (POC). Dual task techniques cover several procedures. Their common feature is that subjects are requested to perform two tasks simultaneously. The crucial hypothesis is: the less an additional or secondary task in the dual task situation will deteriorate in comparison with the base-line single task situation, the lower the mental capacity requirements of the primary task, and vice versa. The approach is now broadened and various versions of task interference under dual task conditions are investigated. For example, the subjects are instructed to perform two tasks concurrently with graded variations of the priorities of the tasks. The POC curve graphically illustrates the effects of possible dual-task combinations arising from sharing limited resources among the concurrently performed tasks.

The critical assumptions of the approach mainly consist in the suggestions that every task will require a certain share of a stable, limited conscious (versus unconscious, automated, implicit or tacit) processing capacity, in the hypothetical additive relationship of the two capacity requirements, and in the restriction of the approach to performance data only. The latter might be misleading for several reasons. First of all there are substantial differences in the sensitivity of performance data and subjectively perceived data. Perceived load seems to be determined mainly by the amount of required resources, often operationalized in terms of working memory, whereas performance measures seem to be determined predominantly by the efficiency of the sharing of resources, depending on allocation strategies (this is dissociation theory; see Wickens and Yeh 1983). Moreover, individual differences in information processing abilities and personality traits strongly influence the indicators of MWL within the subjective (perceived), performance and psychophysiological areas.

Theoretical Approaches: Task Requirement Approaches

As has been shown, task requirements are multidimensional and, thus, may not be described sufficiently by means of only one dimension, whether it be the perceived effort or the residual conscious mental capacity. A more profound description might be a profile-like one, applying a theoretically selected pattern of graded dimensions of task characteristics. The central issue is thus the conceptualization of “task”, especially in terms of task content, and of “task accomplishment”, especially in terms of the structure and phases of goal-oriented actions. The role of the task is stressed by the fact that even the impact of contextual conditions (like temperature, noise or working hours) on the persons are task-dependent, since they are mediated by the task acting as a gate device (Fisher 1986). Various theoretical approaches sufficiently agree regarding those critical task dimensions, which offer a valid prediction of the task outcome. In any case, task outcome is twofold, since (1) the intended result must be achieved, meeting the performance-outcome criteria, and (2) a number of unintended personal short-term and cumulative long-term side effects will emerge, for example fatigue, boredom (monotony), occupational diseases or improved intrinsic motivation, knowledge or skills.

Assessment of MWL. With task requirement approaches, action-oriented approaches like those of complete versus partialized actions or the motivation potential score (for an elaboration of both see Hacker 1986), propose as indispensable task characteristics for analysis and evaluation at least the following:

• temporal and procedural autonomy regarding decisions on self-set goals and, consequently, transparency, predictability and control of the work situation

• number and variety of subtasks (especially concerning preparation, organization and checking of the results of implementation) and of actions accomplishing these subtasks (i.e., whether such actions involve cyclical completeness versus fragmentation)

• variety (“level”) of action-regulating mental processes and representations. These may be mentally automated or routinized ones, knowledge-based “if-then” ones or intellectual and problem-solving ones. (They may also be characterized by hierarchical completeness as opposed to fragmentation)

• required cooperation

• long-term learning requirements or options.

The identification of these task characteristics requires the joint procedures of job/task analysis, including document analyses, observations, interviews and group discussions, which must be integrated in a quasi-experimental design (Rudolph, Schönfelder and Hacker 1987). Task analysis instruments which may guide and assist the analysis are available. Some of them assist only the analysis (for example, NASA-TLX Task Load Index, Hart and Staveland, 1988) while others are useful for evaluation and design or redesign. An example here is the TBS-GA (Tätigkeitsbewertungs System für geistige Arbeit [Task Diagnosis Survey—Mental Work]); see Rudolph, Schönfelder and Hacker (1987).

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The word biomarker is short for biological marker, a term that refers to a measurable event occurring in a biological system, such as the human body. This event is then interpreted as a reflection, or marker, of a more general state of the organism or of life expectancy. In occupational health, a biomarker is generally used as an indicator of health status or disease risk.

Biomarkers are used for in vitro as well as in vivo studies that may include humans. Usually, three specific types of biological markers are identified. Although a few biomarkers may be difficult to classify, usually they are separated into biomarkers of exposure, biomarkers of effect or biomarkers of susceptibility (see table 1).

Table 1. Examples of biomarkers of exposure or biomarkers of effect  that are used in toxicological studies in occupational health

Given an acceptable degree of validity, biomarkers may be employed for several purposes. On an individual basis, a biomarker may be used to support or refute a diagnosis of a particular type of poisoning or other chemically-induced adverse effect. In a healthy subject, a biomarker may also reflect individual hypersusceptibility to specific chemical exposures and may therefore serve as a basis for risk prediction and counselling. In groups of exposed workers, some exposure biomarkers can be applied to assess the extent of compliance with pollution abatement regulations or the effectiveness of preventive efforts in general.

Biomarkers of Exposure

An exposure biomarker may be an exogenous compound (or a metabolite) within the body, an interactive product between the compound (or metabolite) and an endogenous component, or another event related to the exposure. Most commonly, biomarkers of exposures to stable compounds, such as metals, comprise measurements of the metal concentrations in appropriate samples, such as blood, serum or urine. With volatile chemicals, their concentration in exhaled breath (after inhalation of contamination-free air) may be assessed. If the compound is metabolized in the body, one or more metabolites may be chosen as a biomarker of the exposure; metabolites are often determined in urine samples.

Modern methods of analysis may allow separation of isomers or congeners of organic compounds, and determination of the speciation of metal compounds or isotopic ratios of certain elements. Sophisticated analyses allow determination of changes in the structure of DNA or other macromolecules caused by binding with reactive chemicals. Such advanced techniques will no doubt gain considerably in importance for applications in biomarker studies, and lower detection limits and better analytical validity are likely to make these biomarkers even more useful.

Particularly promising developments have occurred with biomarkers of exposure to mutagenic chemicals. These compounds are reactive and may form adducts with macromolecules, such as proteins or DNA. DNA adducts may be detected in white blood cells or tissue biopsies, and specific DNA fragments may be excreted in the urine. For example, exposure to ethylene oxide results in reactions with DNA bases, and, after excision of the damaged base, N-7-(2-hydroxyethyl)guanine will be eliminated in the urine. Some adducts may not refer directly to a particular exposure. For example, 8-hydroxy-2´-deoxyguanosine reflects oxidative damage to DNA, and this reaction may be triggered by several chemical compounds, most of which also induce lipid peroxidation.

Other macromolecules may also be changed by adduct formation or oxidation. Of special interest, such reactive compounds may generate haemoglobin adducts that can be determined as biomarkers of exposure to the compounds. The advantage is that ample amounts of haemoglobin can be obtained from a blood sample, and, given the four-month lifetime of red blood cells, the adducts formed with the amino acids of the protein will indicate the total exposure during this period.

Adducts may be determined by sensitive techniques such as high-performance lipid chromatography, and some immunological methods are also available. In general, the analytical methods are new, expensive and need further development and validation. Better sensitivity can be obtained by using the ³²P post labelling assay, which is a nonspecific indication that DNA damage has taken place. All of these techniques are potentially useful for biological monitoring and have been applied in a growing number of studies. However, simpler and more sensitive analytical methods are needed. Given the limited specificity of some methods at low-level exposures, tobacco smoking or other factors may impact significantly on the measurement results, thus causing difficulties in interpretation.

Exposure to mutagenic compounds, or to compounds which are metabolized into mutagens, may also be determined by assessing the mutagenicity of the urine from an exposed individual. The urine sample is incubated with a strain of bacteria in which a specific point mutation is expressed in a way that can be easily measured. If mutagenic chemicals are present in the urine sample, then an increased rate of mutations will occur in the bacteria.

Exposure biomarkers must be evaluated with regard to temporal variation in exposure and the relation to different compartments. Thus, the time frame(s) represented by the biomarker, that is, the extent to which the biomarker measurement reflects past exposure(s) and/or accumulated body burden, must be determined from toxicokinetic data in order to interpret the result. In particular, the degree to which the biomarker indicates retention in specific target organs should be considered. Although blood samples are often used for biomarker studies, peripheral blood is generally not regarded as a compartment as such, although it acts as a transport medium between compartments. The degree to which the concentration in the blood reflects levels in different organs varies widely between different chemicals, and usually also depends upon the length of the exposure as well as time since exposure.

Sometimes this type of evidence is used to classify a biomarker as an indicator of (total) absorbed dose or an indicator of effective dose (i.e., the amount that has reached the target tissue). For example, exposure to a particular solvent may be evaluated from data on the actual concentration of the solvent in the blood at a particular time following the exposure. This measurement will reflect the amount of the solvent that has been absorbed into the body. Some of the absorbed amount will be exhaled due to the vapour pressure of the solvent. While circulating in the blood, the solvent will interact with various components of the body, and it will eventually become subject to breakdown by enzymes. The outcome of the metabolic processes can be assessed by determining specific mercapturic acids produced by conjugation with glutathione. The cumulative excretion of mercapturic acids may better reflect the effective dose than will the blood concentration.

Life events, such as reproduction and senescence, may affect the distribution of a chemical. The distribution of chemicals within the body is significantly affected by pregnancy, and many chemicals may pass the placental barrier, thus causing exposure of the foetus. Lactation may result in excretion of lipid-soluble chemicals, thus leading to a decreased retention in the mother along with an increased uptake by the infant. During weight loss or development of osteoporosis, stored chemicals may be released, which can then result in a renewed and protracted “endogenous” exposure of target organs. Other factors may affect individual absorption, metabolism, retention and distribution of chemical compounds, and some biomarkers of susceptibility are available (see below).

Biomarkers of Effect

A marker of effect may be an endogenous component, or a measure of the functional capacity, or some other indicator of the state or balance of the body or organ system, as affected by the exposure. Such effect markers are generally preclinical indicators of abnormalities.

These biomarkers may be specific or non-specific. The specific biomarkers are useful because they indicate a biological effect of a particular exposure, thus providing evidence that can potentially be used for preventive purposes. The non-specific biomarkers do not point to an individual cause of the effect, but they may reflect the total, integrated effect due to a mixed exposure. Both types of biomarkers may therefore be of considerable use in occupational health.

There is not a clear distinction between exposure biomarkers and effect biomarkers. For example, adduct formation could be said to reflect an effect rather than the exposure. However, effect biomarkers usually indicate changes in the functions of cells, tissues or the total body. Some researchers include gross changes, such as an increase in liver weight of exposed laboratory animals or decreased growth in children, as biomarkers of effect. For the purpose of occupational health, effect biomarkers should be restricted to those that indicate subclinical or reversible biochemical changes, such as inhibition of enzymes. The most frequently used effect biomarker is probably inhibition of cholinesterase caused by certain insecticides, that is, organophosphates and carbamates. In most cases, this effect is entirely reversible, and the enzyme inhibition reflects the total exposure to this particular group of insecticides.

Some exposures do not result in enzyme inhibition but rather in increased activity of an enzyme. This is the case with several enzymes that belong to the P450 family (see “Genetic determinants of toxic response”). They may be induced by exposures to certain solvents and polyaromatic hydrocarbons (PAHs). Since these enzymes are mainly expressed in tissues from which a biopsy may be difficult to obtain, the enzyme activity is determined indirectly in vivo by administering a compound that is metabolized by that particular enzyme, and then the breakdown product is measured in urine or plasma.

Other exposures may induce the synthesis of a protective protein in the body. The best example is probably metallothionein, which binds cadmium and promotes the excretion of this metal; cadmium exposure is one of the factors that result in increased expression of the metallothionein gene. Similar protective proteins may exist but have not yet been explored sufficiently to become accepted as biomarkers. Among the candidates for possible use as biomarkers are the so-called stress proteins, originally referred to as heat shock proteins. These proteins are generated by a range of different organisms in response to a variety of adverse exposures.

Oxidative damage may be assessed by determining the concentration of malondialdehyde in serum or the exhalation of ethane. Similarly, the urinary excretion of proteins with a small molecular weight, such as albumin, may be used as a biomarker of early kidney damage. Several parameters routinely used in clinical practice (for example, serum hormone or enzyme levels) may also be useful as biomarkers. However, many of these parameters may not be sufficiently sensitive to detect early impairment.

Another group of effect parameters relate to genotoxic effects (changes in the structure of chromosomes). Such effects may be detected by microscopy of white blood cells that undergo cell division. Serious damage to the chromosomes—chromosomal aberrations or formation of micronuclei—can be seen in a microscope. Damage may also be revealed by adding a dye to the cells during cell division. Exposure to a genotoxic agent can then be visualized as an increased exchange of the dye between the two chromatids of each chromosome (sister chromatid exchange). Chromosomal aberrations are related to an increased risk of developing cancer, but the significance of an increased rate of sister chromatid exchange is less clear.

More sophisticated assessment of genotoxicity is based on particular point mutations in somatic cells, that is, white blood cells or epithelial cells obtained from the oral mucosa. A mutation at a specific locus may make the cells capable of growing in a culture that contains a chemical that is otherwise toxic (such as 6-thioguanine). Alternatively, a specific gene product can be assessed (e.g., serum or tissue concentrations of oncoproteins encoded by particular oncogenes). Obviously, these mutations reflect the total genotoxic damage incurred and do not necessarily indicate anything about the causative exposure. These methods are not yet ready for practical use in occupational health, but rapid progress in this line of research would suggest that such methods will become available within a few years.

Biomarkers of Susceptibility

A marker of susceptibility, whether inherited or induced, is an indicator that the individual is particularly sensitive to the effect of a xenobiotic or to the effects of a group of such compounds. Most attention has been focused on genetic susceptibility, although other factors may be at least as important. Hypersusceptibility may be due to an inherited trait, the constitution of the individual, or environmental factors.

The ability to metabolize certain chemicals is variable and is genetically determined (see “Genetic determinants of toxic response”). Several relevant enzymes appear to be controlled by a single gene. For example, oxidation of foreign chemicals is mainly carried out be a family of enzymes belonging to the P450 family. Other enzymes make the metabolites more water soluble by conjugation (e.g., N-acetyltransferase and μ-glutathion-S-transferase). The activity of these enzymes is genetically controlled and varies considerably. As mentioned above, the activity can be determined by administering a small dose of a drug and then determining the amount of the metabolite in the urine. Some of the genes have now been characterized, and techniques are available to determine the genotype. Important studies suggest that a risk of developing certain cancer forms is related to the capability of metabolizing foreign compounds. Many questions still remain unanswered, thus at this time limiting the use of these potential susceptibility biomarkers in occupational health.

Other inherited traits, such as alpha₁-antitrypsin deficiency or glucose-6-phosphate dehydrogenase deficiency, also result in deficient defence mechanisms in the body, thereby causing hypersusceptibility to certain exposures.

Most research related to susceptibility has dealt with genetic predisposition. Other factors play a role as well and have been partly neglected. For example, individuals with a chronic disease may be more sensitive to an occupational exposure. Also, if a disease process or previous exposure to toxic chemicals has caused some subclinical organ damage, then the capacity to withstand a new toxic exposure is likely to be less. Biochemical indicators of organ function may in this case be used as susceptibility biomarkers. Perhaps the best example regarding hypersusceptibility relates to allergic responses. If an individual has become sensitized to a particular exposure, then specific antibodies can be detected in serum. Even if the individual has not become sensitized, other current or past exposures may add to the risk of developing an adverse effect related to an occupational exposure.

A major problem is to determine the joint effect of mixed exposures at work. In addition, personal habits and drug use may result in an increased susceptibility. For example, tobacco smoke usually contains a considerable amount of cadmium. Thus, with occupational exposure to cadmium, a heavy smoker who has accumulated substantial amounts of this metal in the body will be at increased risk of developing cadmium-related kidney disease.

Application in Occupational Health

Biomarkers are extremely useful in toxicological research, and many may be applicable in biological monitoring. Nonetheless, the limitations must also be recognized. Many biomarkers have so far been studied only in laboratory animals. Toxicokinetic patterns in other species may not necessarily reflect the situation in human beings, and extrapolation may require confirmatory studies in human volunteers. Also, account must be taken of individual variations due to genetic or constitutional factors.

In some cases, exposure biomarkers may not at all be feasible (e.g., for chemicals which are short-lived in vivo). Other chemicals may be stored in, or may affect, organs which cannot be accessed by routine procedures, such as the nervous system. The route of exposure may also affect the distribution pattern and therefore also the biomarker measurement and its interpretation. For example, direct exposure of the brain via the olfactory nerve is likely to escape detection by measurement of exposure biomarkers. As to effect biomarkers, many of them are not at all specific, and the change can be due to a variety of causes, including lifestyle factors. Perhaps in particular with the susceptibility biomarkers, interpretation must be very cautious at the moment, as many uncertainties remain about the overall health significance of individual genotypes.

In occupational health, the ideal biomarker should satisfy several requirements. First of all, sample collection and analysis must be simple and reliable. For optimal analytical quality, standardization is needed, but the specific requirements vary considerably. Major areas of concern include: preparation of the in- dividual, sampling procedure and sample handling, and measurement procedure; the latter encompasses technical factors, such as calibration and quality assurance procedures, and individual- related factors, such as education and training of operators.

For documentation of analytical validity and traceability, reference materials should be based on relevant matrices and with appropriate concentrations of toxic substances or relevant metabolites at appropriate levels. For biomarkers to be used for biological monitoring or for diagnostic purposes, the responsible laboratories must have well-documented analytical procedures with defined performance characteristics, and accessible records to allow verification of the results. At the same time, nonetheless, the economics of characterizing and using reference materials to supplement quality assurance procedures in general must be considered. Thus, the achievable quality of results, and the uses to which they are put, have to be balanced against the added costs of quality assurance, including reference materials, manpower and instrumentation.

Another requirement is that the biomarker should be specific, at least under the circumstances of the study, for a particular type of exposure, with a clear-cut relationship to the degree of exposure. Otherwise, the result of the biomarker measurement may be too difficult to interpret. For proper interpretation of the measurement result of an exposure biomarker, the diagnostic validity must be known (i.e., the translation of the biomarker value into the magnitude of possible health risks). In this area, metals serve as a paradigm for biomarker research. Recent research has demonstrated the complexity and subtlety of dose-response relationships, with considerable difficulty in identifying no-effect levels and therefore also in defining tolerable exposures. However, this kind of research has also illustrated the types of investigation and the refinement that are necessary to uncover the relevant information. For most organic compounds, quantitative associations between exposures and the corresponding adverse health effects are not yet available; in many cases, even the primary target organs are not known for sure. In addition, evaluation of toxicity data and biomarker concentrations is often complicated by exposure to mixtures of substances, rather than exposure to a single compound at the time.

Before the biomarker is applied for occupational health purposes, some additional considerations are necessary. First, the biomarker must reflect a subclinical and reversible change only. Second, given that the biomarker results can be interpreted with regard to health risks, then preventive efforts should be available and should be considered realistic in case the biomarker data suggests a need to reduce the exposure. Third, the practical use of the biomarker must be generally regarded as ethically acceptable.

Industrial hygiene measurements may be compared with applicable exposure limits. Likewise, results on exposure biomarkers or effect biomarkers may be compared to biological action limits, sometimes referred to as biological exposure indices. Such limits should be based on the best advice of clinicians and scientists from appropriate disciplines, and responsible administrators as “risk managers” should then take into account relevant ethical, social, cultural and economic factors. The scientific basis should, if possible, include dose-response relationships supplemented by information on variations in susceptibility within the population at risk. In some countries, workers and members of the general public are involved in the standard-setting process and provide important input, particularly when scientific uncertainty is considerable. One of the major uncertainties is how to define an adverse health effect that should be prevented—for example, whether adduct formation as an exposure biomarker by itself represents an adverse effect (i.e., effect biomarker) that should be prevented. Difficult questions are likely to arise when deciding whether it is ethically defensible, for the same compound, to have different limits for adventitious exposure, on the one hand, and occupational exposure, on the other.

The information generated by the use of biomarkers should generally be conveyed to the individuals examined within the physician-patient relationship. Ethical concerns must in particular be considered in connection with highly experimental biomarker analyses that cannot currently be interpreted in detail in terms of actual health risks. For the general population, for example, limited guidance exists at present with regard to interpretation of exposure biomarkers other than the blood-lead concentration. Also of importance is the confidence in the data generated (i.e., whether appropriate sampling has been done, and whether sound quality assurance procedures have been utilized in the laboratory involved). An additional area of special worry relates to individual hypersusceptibility. These issues must be taken into account when providing the feedback from the study.

All sectors of society affected by, or concerned with carrying out, a biomarker study need to be involved in the decision-making process on how to handle the information generated by the study. Specific procedures to prevent or overcome inevitable ethical conflicts should be developed within the legal and social frameworks of the region or country. However, each situation represents a different set of questions and pitfalls, and no single procedure for public involvement can be developed to cover all applications of exposure biomarkers.

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The concept of vigilance refers to a human observer’s state of alertness in tasks that demand efficient registration and processing of signals. The main characteristics of vigilance tasks are relatively long durations and the requirement to detect infrequent and unpredictable target stimuli (signals) against a background of other stimulus events.

Vigilance Tasks

The prototypical task for vigilance research was that of radar operators. Historically, their apparently unsatisfactory performance during the Second World War has been a major impetus for the extensive study of vigilance. Another major task requiring vigilance is an industrial inspection. More generally, all kinds of monitoring tasks which require the detection of relatively infrequent signals embody the risk of failures to detect and to respond to these critical events.

Vigilance tasks make up a heterogeneous set and vary on several dimensions, in spite of their common characteristics. An obviously important dimension is the overall stimulus rate as well as the rate of target stimuli. It is not always possible to define the stimulus rate unambiguously. This is the case in tasks that require the detection of target events against continuously presented background stimuli, as in detecting critical values on a set of dials in a monitoring task. A less obviously important distinction is that between successive-discrimination tasks and simultaneous-discrimination tasks. In simultaneous-discrimination tasks, both target stimuli and background stimuli are present at the same time, while in successive-discrimination tasks one is presented after the other so that some demands on memory are made. Although most vigilance tasks require the detection of visual stimuli, stimuli in other modalities have also been studied. Stimuli can be confined to a single spatial location, or there can be different sources for target stimuli. Target stimuli can differ from background stimuli by physical characteristics, but also by more conceptual ones (like a certain pattern of meter readings that can differ from other patterns). Of course, the conspicuousness of targets can vary: some can be detected easily, while others may be hard to discriminate from background stimuli. Target stimuli can be unique or there can be sets of target stimuli without well-defined boundaries to set them off from background stimuli, as is the case in many industrial inspection tasks. This list of dimensions on which vigilance tasks differ can be expanded, but even this length of the list suffices to emphasize the heterogeneity of vigilance tasks and thus the risks involved in generalizing certain observations across the full set.

Performance Variations and the Vigilance Decrement

The most frequently used performance measure in vigilance tasks is the proportion of target stimuli, for example, faulty products in industrial inspection, that have been detected; this is an estimate of the probability of so-called hits. Those target stimuli that remain unnoticed are called misses. Although the hit rate is a convenient measure, it is somewhat incomplete. There is a trivial strategy that allows one to achieve 100% hits: one only has to classify all stimuli as targets. However, the hit rate of 100% is then accompanied by a false-alarm rate of 100%, that is, not only the target stimuli are correctly detected, but the background stimuli are incorrectly “detected” as well. This line of reasoning makes it quite clear that whenever there are false alarms at all, it is important to know their proportion in addition to the hit rate. Another measure for performance in a vigilance task is the time needed to respond to target stimuli (response time).

Performance in vigilance tasks exhibits two typical attributes. The first one is the low overall level of vigilance performance. It is low in comparison with an ideal situation for the same stimuli (short observation periods, high readiness of the observer for each discrimination, etc.). The second attribute is the so-called vigilance decrement, the decline of performance in the course of the watch which can start within the first few minutes. Both these observations refer to the proportion of hits, but they have also been reported for response times. Although the vigilance decrement is typical of vigilance tasks, it is not universal.

In investigating the causes of poor overall performance and vigilance decrements, a distinction will be made among concepts that are related to the basic characteristics of the task and concepts that are related to organismic and task-unrelated situational factors. Among the task-related factors strategic and non-strategic ones can be distinguished.

Strategic processes in vigilance tasks

The detection of a signal like a faulty product is partly a matter of the observer’s strategy and partly a matter of the signal’s discriminability. This distinction is based on the theory of signal detection (TSD), and some basics of the theory need to be presented in order to highlight the distinction’s importance. Consider a hypothetical variable, defined as “evidence for the presence of a signal”. Whenever a signal is presented, this variable takes on some value, and whenever a background stimulus is presented, it takes on a value that is lower on the average. The value of the evidence variable is assumed to vary across repeated presentations of the signal. Thus it can be characterized by a so-called probability density function as is illustrated in figure 1. Another density function characterizes the values of the evidence variable upon presentation of a background stimulus. When the signals are similar to the background stimuli, the functions will overlap, so that a certain value of the evidence variable can originate either from a signal or from a background stimulus. The particular shape of the density functions of figure 1 is not essential for the argument.

Figure 1. Thresholds and discriminability

The detection response of the observer is based on the evidence variable. It is assumed that a threshold is set so that a detection response is given whenever the value of the evidence variable is above the threshold. As is illustrated in figure 1, the areas under the density functions to the right of the threshold correspond to the probabilities of hits and false alarms. In practice, estimates of the separation of the two functions and the location of the threshold can be derived. The separation of the two density functions characterizes the discriminability of the target stimuli from the background stimuli, while the location of the threshold characterizes the observer’s strategy. Variation of the threshold produces a joint variation of the proportions of hits and false alarms. With a high threshold, the proportions of hits and false alarms will be small, while with a low threshold the proportions will be large. Thus, the selection of a strategy (placement of the threshold) essentially is the selection of a certain combination of hit rate and false-alarm rate among the combinations that are possible for a certain discriminability.

Two major factors that influence the location of the threshold are payoffs and signal frequency. The threshold will be set to lower values when there is much to gain from a hit and little to lose from a false alarm, and it will be set to higher values when false alarms are costly and the benefits from hits are small. A low threshold setting can also be induced by a high proportion of signals, while a low proportion of signals tends to induce higher threshold settings. The effect of signal frequency on threshold settings is a major factor for the low overall performance in terms of the proportion of hits in vigilance tasks and for the vigilance decrement.

An account of the vigilance decrement in terms of strategic changes (threshold changes) requires that the reduction of the proportion of hits in the course of the watch is accompanied by a reduction of the proportion of false alarms. This is, in fact, the case in many studies, and it is likely that the overall poor performance in vigilance tasks (in comparison with the optimal situation) does also result, at least partly, from a threshold adjustment. In the course of a watch, the relative frequency of detection responses comes to match the relative frequency of targets, and this adjustment implies a high threshold with a relatively small proportion of hits and a relatively small proportion of false alarms as well. Nevertheless, there are vigilance decrements that result from changes in discriminability rather than from changes in threshold settings. These have been observed mainly in successive-discrimination tasks with a relatively high rate of stimulus events.

Nonstrategic processes in vigilance tasks

Although part of the overall poor performance in vigilance tasks and many instances of the vigilance decrement can be accounted for in terms of strategic adjustments of the detection threshold to low signal rates, such an account is not complete. There are changes in the observer during a watch that can reduce the discriminability of stimuli or result in apparent threshold shifts that cannot be considered as an adaptation to the task characteristics. In the more than 40 years of vigilance research, a number of nonstrategic factors that contribute to poor overall performance and to the vigilance decrement have been identified.

A correct response to a target in a vigilance task requires a sufficiently precise sensory registration, an appropriate threshold location, and a link between the perceptual processes and the associated response-related processes. During the watch the observers have to maintain a certain task set, a certain readiness to respond to target stimuli in a certain way. This is a nontrivial requirement because without a particular task set no observer would respond to target stimuli in the way required. Two major sources of failures are thus inaccurate sensory registration and lapses in the readiness to respond to target stimuli. Major hypotheses to account for such failures will be briefly reviewed.

Detection and identification of a stimulus are faster when there is no temporal or spatial uncertainty about its appearance. Temporal and/or spatial uncertainty is likely to reduce vigilance performance. This is the essential prediction of expectancy theory. Optimal preparedness of the observer requires temporal and spatial certainty; obviously vigilance tasks are less than optimal in this respect. Although the major focus of expectancy theory is on the overall low performance, it can also serve to account for parts of the vigilance decrement. With infrequent signals at random intervals, high levels of preparedness might initially exist at times when no signal is presented; in addition, signals will be presented at low levels of preparedness. This discourages occasional high levels of preparedness in general so that whatever benefits accrue from them will vanish in the course of a watch.

Expectancy theory has a close relation to attentional theories. Variants of attentional theories of vigilance, of course, are related to dominant theories of attention in general. Consider a view of attention as “selection for processing” or “selection for action”. According to this view, stimuli are selected from the environment and processed with high efficiency whenever they serve the currently dominant action plan or task set. As already said, the selection will benefit from precise expectations about when and where such stimuli will occur. But stimuli will only be selected if the action plan—the task set—is active. (Drivers of cars, for example, respond to traffic lights, other traffic, etc.; passengers don’t do so normally, although both are in almost the same situation. The critical difference is that between the task sets of the two: only the driver’s task set requires responses to traffic lights.)

The selection of stimuli for processing will suffer when the action plan is temporarily deactivated, that is when the task set is temporarily absent. Vigilance tasks embody a number of features that discourage continuous maintenance of the task set, like short cycle times for processing stimuli, lack of feedback and little motivational challenge by apparent task difficulty. So-called blockings can be observed in almost all simple cognitive tasks with short cycle times like simple mental arithmetic or rapid serial responses to simple signals. Similar blockings occur in the maintenance of the task set in a vigilance task as well. They are not immediately recognizable as delayed responses because responses are infrequent and targets that are presented during a period of absent task set may no longer be there when the absence is over so that a miss will be observed instead of a delayed response. Blockings become more frequent with time spent on the task. This can give rise to the vigilance decrement. There may be additional reasons for temporary lapses in the availability of the appropriate task set, for example, distraction.

Certain stimuli are not selected in the service of the current action plan, but by virtue of their own characteristics. These are stimuli that are intense, novel, moving toward the observer, have an abrupt onset or for any other reason might require immediate action no matter what the current action plan of the observer is. There is little risk of not detecting such stimuli. They attract attention automatically, as is indicated, for example, by the orienting response, which includes a shift of the direction of the gaze toward the stimulus source. However, answering an alarm bell is not normally considered a vigilance task. In addition to stimuli that attract attention by their own characteristics, there are stimuli that are processed automatically as a consequence of the practice. They seem to “pop out” from the environment. This kind of automatic processing requires extended practice with a so-called consistent mapping, that is, a consistent assignment of responses to stimuli. The vigilance decrement is likely to be small or even absent once automatic processing of stimuli has been developed.

Finally, vigilance performance suffers from a lack of arousal. This concept refers in a rather global manner to the intensity of neural activity, ranging from sleep through normal wakefulness to high excitement. One of the factors that is thought to affect arousal is external stimulation, and this is fairly low and uniform in most vigilance tasks. Thus, the intensity of central nervous system activity can decline overall over the course of a watch. An important aspect of arousal theory is that it links vigilance performance to various task-unrelated situational factors and factors related to the organism.

The Influence of Situational and Organismic Factors

Low arousal contributes to poor performance in vigilance tasks. Thus performance can be enhanced by situational factors that tend to enhance arousal, and it can be reduced by all measures that reduce the level of arousal. On balance, this generalization is mostly correct for the overall performance level in vigilance tasks, but the effects on the vigilance decrement are absent or less reliably observed across different kinds of manipulation of arousal.

One way to raise the level of arousal is the introduction of additional noise. However, the vigilance decrement is generally unaffected, and with respect to overall performance the results are inconsistent: enhanced, unchanged and reduced performance levels have all been observed. Perhaps the complex nature of noise is relevant. For example, it can be affectively neutral or annoying; it cannot only be arousing, but also be distracting. More consistent are the effects of sleep deprivation, which is “de-arousing”. It generally reduces vigilance performance and has sometimes been seen to enhance the vigilance decrement. Appropriate changes of vigilance performance have also been observed with depressant drugs like benzodiazepines or alcohol and stimulant drugs like amphetamine, caffeine or nicotine.

Individual differences are a conspicuous feature of performance in vigilance tasks. Although individual differences are not consistent across all sorts of vigilance tasks, they are fairly consistent across similar ones. There is only little or no effect of sex and general intelligence. With respect to age, vigilance performance increases during childhood and tends to decline beyond the age of sixty. In addition there is a good chance that introverts will show better performance than extroverts.

The Enhancement of Vigilance Performance

The existing theories and data suggest some means to enhance vigilance performance. Depending on the amount of specificity of the suggestions, it is not difficult to compile lists of various lengths. Some rather broad suggestions are given below that have to be fitted to specific task requirements. They are related to the ease of perceptual discriminations, the appropriate strategic adjustments, the reduction of uncertainty, the avoidance of the effects of attentional lapses and the maintenance of arousal.

Vigilance tasks require discriminations under non-optimal conditions. Thus one is well advised in making the discriminations as easy as possible, or the signals as conspicuous as possible. Measures related to this general goal can be straightforward (like appropriate lighting or longer inspection times per product) or more sophisticated, including special devices to enhance the conspicuousness of targets. Simultaneous comparisons are easier than successive ones, so the availability of a reference standard can be helpful. By means of technical devices, it is sometimes possible to present the standard and the object to be examined in rapid alternation, so that differences will appear as motions in the display or other changes for which the visual system is particularly sensitive.

To counteract the strategic changes of the threshold that lead to a relatively low proportion of correct detections of targets (and for making the task less boring in terms of the frequency of actions to be taken) the suggestion has been made to introduce fake targets. However, this seems not to be a good recommendation. Fake targets will increase the proportion of hits overall but at the cost of more frequent false alarms. In addition, the proportion of undetected targets among all stimuli that are not responded to (the outgoing faulty material in an industrial inspection task) will not necessarily be reduced. Better suited seems to be explicit knowledge about the relative importance of hits and false alarms and perhaps other measures to obtain an appropriate placement of the threshold for deciding between “good” and “bad”.

Temporal and spatial uncertainty are important determinants of poor vigilance performance. For some tasks, spatial uncertainty can be reduced by way of defining a certain position of the object to be inspected. However, little can be done about temporal uncertainty: the observer would be unnecessary in a vigilance task if the occurrence of a target could be signaled in advance of its presentation. One thing that can be done in principle, however, is to mix objects to be inspected if faults tend to occur in bunches; this serves to avoid very long intervals without targets as well as very short intervals.

There are some obvious suggestions for the reduction of attentional lapses or at least their impact on performance. By proper training, some kind of automatic processing of targets can perhaps be obtained provided that the background and target stimuli are not too variable. The requirement for sustained maintenance of the task set can be avoided by means of frequent short breaks, job rotation, job enlargement or job enrichment. Introduction of variety can be as simple as having the inspector himself or herself getting the material to be inspected from a box or other location. This also introduces self-pacing, which may help in avoiding signal presentations during temporary deactivations of the task set. Sustained maintenance of task set can be supported by means of feedback, indicated interest by supervisors and operator’s awareness of the importance of the task. Of course, accurate feedback of performance level is not possible in typical vigilance tasks; however, even inaccurate or incomplete feedback can be helpful as far as the observer’s motivation is concerned.

There are some measures that can be taken to maintain a sufficient level of arousal. Continuous use of drugs may exist in practice but is never found among recommendations. Some background music can be useful, but can also have an opposite effect. Social isolation during vigilance tasks should mostly be avoided, and during times of day with low levels of arousal like the late hours of the night, supportive measures such as short watches are particularly important.

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Genetic toxicity assessment is the evaluation of agents for their ability to induce any of three general types of changes (mutations) in the genetic material (DNA): gene, chromosomal and genomic. In organisms such as humans, the genes are composed of DNA, which consists of individual units called nucleotide bases. The genes are arranged in discrete physical structures called chromosomes. Genotoxicity can result in significant and irreversible effects upon human health. Genotoxic damage is a critical step in the induction of cancer and it can also be involved in the induction of birth defects and foetal death. The three classes of mutations mentioned above can occur within either of the two types of tissues possessed by organisms such as humans: sperm or eggs (germ cells) and the remaining tissue (somatic cells).

Assays that measure gene mutation are those that detect the substitution, addition or deletion of nucleotides within a gene. Assays that measure chromosomal mutation are those that detect breaks or chromosomal rearrangements involving one or more chromosomes. Assays that measure genomic mutation are those that detect changes in the number of chromosomes, a condition called aneuploidy. Genetic toxicity assessment has changed considerably since the development by Herman Muller in 1927 of the first assay to detect genotoxic (mutagenic) agents. Since then, more than 200 assays have been developed that measure mutations in DNA; however, fewer than ten assays are used commonly today for genetic toxicity assessment. This article reviews these assays, describes what they measure, and explores the role of these assays in toxicity assessment.

Identification of Cancer HazardsPrior to the Development of the Fieldof Genetic Toxicology

Genetic toxicology has become an integral part of the overall risk assessment process and has gained in stature in recent times as a reliable predictor for carcinogenic activity. However, prior to the development of genetic toxicology (before 1970), other methods were and are still being used to identify potential cancer hazards to humans. There are six major categories of methods currently used for identifying human cancer risks: epidemiological studies, long-term in vivo bioassays, mid-term in vivo bioassays, short-term in vivo and in vitro bioassays, artificial intelligence (structure-activity), and mechanism-based inference.

Table 1 gives advantages and disadvantages for these methods.

Table 1. Advantages and disadvantages of current methods for identifying human cancer risks

Rationale and Conceptual Basisfor Genetic Toxicology Assays

Although the exact types and numbers of assays used for genetic toxicity assessment are constantly evolving and vary from country to country, the most common ones include assays for (1) gene mutation in bacteria and/or cultured mammalian cells and (2) chromosomal mutation in cultured mammalian cells and/or bone marrow within living mice. Some of the assays within this second category can also detect aneuploidy. Although these assays do not detect mutations in germ cells, they are used primarily because of the extra cost and complexity of performing germ-cell assays. Nonetheless, germ-cell assays in mice are used when information about germ-cell effects is desired.

Systematic studies over a 25-year period (1970-1995), especially at the US National Toxicology Program in North Carolina, have resulted in the use of a discrete number of assays for detecting the mutagenic activity of agents. The rationale for evaluating the usefulness of the assays was based on their ability to detect agents that cause cancer in rodents and that are suspected of causing cancer in humans (i.e., carcinogens). This is because studies during the past several decades have indicated that cancer cells contain mutations in certain genes and that many carcinogens are also mutagens. Thus, cancer cells are viewed as containing somatic-cell mutations, and carcinogenesis is viewed as a type of somatic-cell mutagenesis.

The genetic toxicity assays used most commonly today have been selected not only because of their large database, relatively low cost, and ease of performance, but because they have been shown to detect many rodent and, presumptively, human carcinogens. Consequently, genetic toxicity assays are used to predict the potential carcinogenicity of agents.

An important conceptual and practical development in the field of genetic toxicology was the recognition that many carcinogens were modified by enzymes within the body, creating altered forms (metabolites) that were frequently the ultimate carcinogenic and mutagenic form of the parent chemical. To duplicate this metabolism in a petri dish, Heinrich Malling showed that the inclusion of a preparation from rodent liver contained many of the enzymes necessary to perform this metabolic conversion or activation. Thus, many genetic toxicity assays performed in dishes or tubes (in vitro) employ the addition of similar enzyme preparations. Simple preparations are called S9 mix, and purified preparations are called microsomes. Some bacterial and mammalian cells have now been genetically engineered to contain some of the genes from rodents or humans that produce these enzymes, reducing the need to add S9 mix or microsomes.

Genetic Toxicology Assays and Techniques

The primary bacterial systems used for genetic toxicity screening are the Salmonella (Ames) mutagenicity assay and, to a much lesser extent, strain WP2 of Escherichia coli. Studies in the mid-1980s indicated that the use of only two strains of the Salmonella system (TA98 and TA100) were sufficient to detect approximately 90% of the known Salmonella mutagens. Thus, these two strains are used for most screening purposes; however, various other strains are available for more extensive testing.

These assays are performed in a variety of ways, but two general procedures are the plate-incorporation and liquid-suspension assays. In the plate-incorporation assay, the cells, the test chemical and (when desired) the S9 are added together into a liquefied agar and poured onto the surface of an agar petri plate. The top agar hardens within a few minutes, and the plates are incubated for two to three days, after which time mutant cells have grown to form visually detectable clusters of cells called colonies, which are then counted. The agar medium contains selective agents or is composed of ingredients such that only the newly mutated cells will grow. The liquid-incubation assay is similar, except the cells, test agent, and S9 are incubated together in liquid that does not contain liquefied agar, and then the cells are washed free of the test agent and S9 and seeded onto the agar.

Mutations in cultured mammalian cells are detected primarily in one of two genes: hprt and tk. Similar to the bacterial assays, mammalian cell lines (developed from rodent or human cells) are exposed to the test agent in plastic culture dishes or tubes and then are seeded into culture dishes that contain medium with a selective agent that permits only mutant cells to grow. The assays used for this purpose include the CHO/HPRT, the TK6, and the mouse lymphoma L5178Y/TK^+/- assays. Other cell lines containing various DNA repair mutations as well as containing some human genes involved in metabolism are also used. These systems permit the recovery of mutations within the gene (gene mutation) as well as mutations involving regions of the chromosome flanking the gene (chromosomal mutation). However, this latter type of mutation is recovered to a much greater extent by the tk gene systems than by the hprt gene systems due to the location of the tk gene.

Similar to the liquid-incubation assay for bacterial mutagenicity, mammalian cell mutagenicity assays generally involve the exposure of the cells in culture dishes or tubes in the presence of the test agent and S9 for several hours. The cells are then washed, cultured for several more days to allow the normal (wild-type) gene products to be degraded and the newly mutant gene products to be expressed and accumulate, and then they are seeded into medium containing a selective agent that permits only the mutant cells to grow. Like the bacterial assays, the mutant cells grow into visually detectable colonies that are then counted.

Chromosomal mutation is identified primarily by cytogenetic assays, which involve exposing rodents and/or rodent or human cells in culture dishes to a test chemical, allowing one or more cell divisions to occur, staining the chromosomes, and then visually examining the chromosomes through a microscope to detect alterations in the structure or number of chromosomes. Although a variety of endpoints can be examined, the two that are currently accepted by regulatory agencies as being the most meaningful are chromosomal aberrations and a subcategory called micronuclei.

Considerable training and expertise are required to score cells for the presence of chromosomal aberrations, making this a costly procedure in terms of time and money. In contrast, micronuclei require little training, and their detection can be automated. Micronuclei appear as small dots within the cell that are distinct from the nucleus, which contains the chromosomes. Micronuclei result from either chromosome breakage or from aneuploidy. Because of the ease of scoring micronuclei compared to chromosomal aberrations, and because recent studies indicate that agents that induce chromosomal aberrations in the bone marrow of living mice generally induce micronuclei in this tissue, micronuclei are now commonly measured as an indication of the ability of an agent to induce chromosomal mutation.

Although germ-cell assays are used far less frequently than the other assays described above, they are indispensable in determining whether an agent poses a risk to the germ cells, mutations in which can lead to health effects in succeeding generations. The most commonly used germ-cell assays are in mice, and involve systems that detect (1) heritable translocations (exchanges) among chromosomes (heritable translocation assay), (2) gene or chromosomal mutations involving specific genes (visible or biochemical specific-locus assays), and (3) mutations that affect viability (dominant lethal assay). As with the somatic-cell assays, the working assumption with the germ-cell assays is that agents positive in these assays are presumed to be potential human germ-cell mutagens.

Current Status and Future Prospects

Recent studies have indicated that only three pieces of information were necessary to detect approximately 90% of a set of 41 rodent carcinogens (i.e., presumptive human carcinogens and somatic-cell mutagens). These included (1) knowledge of the chemical structure of agent, especially if it contains electrophilic moieties (see section on structure-activity relationships); (2) Salmonella mutagenicity data; and (3) data from a 90-day chronic toxicity assay in rodents (mice and rats). Indeed, essentially all of the IARC-declared human carcinogens are detectable as mutagens using just the Salmonella assay and the mouse-bone marrow micronucleus assay. The use of these mutagenicity assays for detecting potential human carcinogens is supported further by the finding that most human carcinogens are carcinogenic in both rats and mice (trans-species carcinogens) and that most trans- species carcinogens are mutagenic in Salmonella and/or induce micronuclei in mouse bone marrow.

With advances in DNA technology, the human genome project, and an improved understanding of the role of mutation in cancer, new genotoxicity assays are being developed that will likely be incorporated into standard screening procedures. Among these are the use of transgenic cells and rodents. Transgenic systems are those in which a gene from another species has been introduced into a cell or organism. For example, transgenic mice are now in experimental use that permit the detection of mutation in any organ or tissue of the animal, based on the introduction of a bacterial gene into the mouse. Bacterial cells, such as Salmonella, and mammalian cells (including human cell lines) are now available that contain genes involved in the metabolism of carcinogenic/mutagenic agents, such as the P450 genes. Molecular analysis of the actual mutations induced in the trans-gene within transgenic rodents, or within native genes such as hprt, or the target genes within Salmonella can now be performed, so that the exact nature of the mutations induced by the chemicals can be determined, providing insights into the mechanism of action of the chemical and allowing comparisons to mutations in humans presumptively exposed to the agent.

Molecular advances in cytogenetics now permit more detailed evaluation of chromosomal mutations. These include the use of probes (small pieces of DNA) that attach (hybridize) to specific genes. Rearrangements of genes on the chromosome can then be revealed by the altered location of the probes, which are fluorescent and easily visualized as colored sectors on the chromosomes. The single-cell gel electrophoresis assay for DNA breakage (commonly called the “comet” assay) permits the detection of DNA breaks within single cells and may become an extremely useful tool in combination with cytogenetic techniques for detecting chromosomal damage.

After many years of use and the generation of a large and systematically developed database, genetic toxicity assessment can now be done with just a few assays for relatively small cost in a short period of time (a few weeks). The data produced can be used to predict the ability of an agent to be a rodent and, presumptively, human carcinogen/somatic-cell mutagen. Such an ability makes it possible to limit the introduction into the environment of mutagenic and carcinogenic agents and to develop alternative, nonmutagenic agents. Future studies should lead to even better methods with greater predictivity than the current assays.

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