**1. Introduction**

184 Novel Approaches and Their Applications in Risk Assessment

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Health hazard from natural and anthropogenic sources has begun to be analyzed 2-3 decades ago. Until recently this analysis was primarily applied for human safety in incidents. However, due to stochastic features of risk assessment its applicability to the diagnosis, prevention, and protective or compensatory measures in some cases can be very limited.

The proposed concept combining approaches based on risk analysis and sanogenetic analysis is aimed at increasing the efficiency of both approaches in addressing issues of risk prediction.

There are several reasons necessitating integrated approach to risk analysis:


Polysystemic Approach to Risk Assessment 187


Individual *functional sufficiency of the cardiorespiratory system* was evaluated using a Spiroarteriocardiorhythmograph instrument complex (SACR, recommended by Ministry of Health Care and Social Development of the Russian Federation for clinical use; registration certificate #29/03020703/5869-04, St. Petersburg) allowing simultaneous recording of the heart, vascular, and respiratory rhythms. The method makes it possible to calculate the relative contribution of sympathetic and parasympathetic autonomic nervous system (ANS) into heart rate and BP regulation, integrated values of cardiogram intervals, parameters of

Electrocardiogram (ECG) was recorded in standard lead I over 2 minutes. The timeamplitude parameters of PQRST complex and heart rhythm variability (HRV) were evaluated using statistic, geometric, and spectral parameters. HRV power in different frequency bands determined using Fourier-transform analysis characterizes ANS activity and the function of the central mechanisms of heart rate regulation. Three frequency bands can be distinguished in spectra: very low frequency (VLF, 0-0.04 Hz), low frequency (LF, 0.04-0.15 Hz), and high frequency (HF 0.15-0.4 Hz), which are measured in absolute values of power (msec2). These values can also be presented in standardized units (LFn, HFn) calculated as the ratio of each spectral component to their sum. Index of autonomic balance (AB=LF/HF) and index of centralization (C=(VLF+LF)/HF) were calculated from HRV

Peripheral systolic and diastolic blood pressure (SBP and DBP, respectively) and their variability were measured on middle phalanx using the method of Penaz. From the parameters of BP pulse wave, hemodynamic parameters, stroke volume, and cardiac output were calculated using phase analysis of cardiac cycle and BP. Spontaneous arterial baroreflex sensitivity (BRS=LFM/LFSBP) was also evaluated. From geometric parameters of HRV (mode, mode amplitude, amplitude of oscillations, etc.), autonomic balance index, parameter of adequacy of regulation processes, autonomic rhythm index, and regulatory

For evaluation of functional reserves of the cardiovascular system, a functional test with increased "dead space" was used (Trukhanov et al., 2007). Reactivity of the cardiovascular system was evaluated by changes in the parameters describing its function (in %) during ECG recording in spirometric mask in comparison to ECG recorded without the mask. The time of inspiration and expiration, volume rate of inspiration and expiration, and respiratory volume of quiet breathing in an averaged cycle were evaluated. Parameters of forced expiration (vital capacity of the lungs and volume of forced expiration) were also

The study of the *latent period of simple sensorimotor reaction and other psychomotor parameters* was performed using a specific instrument called "CMM" (computer movement meter), Registration Certificate # 29/03041202/5085-03. The accuracy of measuring the time of simple sensorimotor reaction was 1 ms. A subject was placed in a comfortable chair,

All tests were performed with strict adherence of general bioethical standards.

metabolic and immune processes.

lung ventilation, baroreflex parameters, etc.

system strain index (SI) were calculated.

spectral parameters.

measured.

chemical, radiation, climatic, noise, vibration, and other impacts. Hence, they do not provide a reliable argument for choosing optimal preventive measures.

7. The method of documentation of pathological consequences rules out the possibility of early prophylactic protection of the population contacting with hazardous industries, which is a priori more efficient and less costly than treatment of the realized risks.

By the wide range of effects, all existing real dangers of radiation, physical, chemical, and biological nature can be divided into two categories: 1) risks in the deterministic range of doses and concentrations (doses and concentrations far surpassing than the established thresholds), 2) risks in a stochastic range of doses and concentrations (doses and concentration near the established thresholds). In the deterministic range of doses and concentrations, the biological effects strictly depend on doses and concentrations of anthropogenic factors and can be detected by existing methods of epidemiological analysis. In the stochastic range of doses and concentrations of anthropogenic factors, the consequences strictly depend on individual sensitivity of biological objects, including humans.

In the human body, many regulatory systems operating at different levels of organization provide sensitivity or resistance to both external and internal factors. On the results of analysis of functional adequacy of sanogenesis systems predicts the level of resistance or sensitivity to relatively tolerable doses and concentrations of anthropogenic influences. Hence, sanogenetic monitoring is more informative for the range of doses and concentrations that cause stochastic effects.

Due to individual variability of functioning of sanogenetic processes, the same anthropogenic factor in equal doses and concentrations will cause certain effects in some organisms (sensitive), will not cause in others, and will induce resistance in the third. This implies that at the population level three subpopulations should be determined at relatively low-dose and low-concentration exposures: sensitive, neutral, and super resistant. The ratio between these subpopulations can eventually serve as a criterion of population risk from this exposure.

Any stable fixation of the pathological trace is preceded by processes of dysregulation of the corresponding functions. The most probable pathological outcomes can be predicted on the basis of the results of polysystemic sanogenetic monitoring by detecting dysregulation in certain systems of the organism (cardiorespiratory, psychomotor, and metabolism systems). Monitoring is carried out using computerized measurement instrumentation and data processing systems, which provides the basis for strict quantitative assessment of the dynamics of risk for the studied populations. The risks assessment goes from the instrument of control to the rank of controlled processes, which is the basis for successful operation of potentially hazardous industries.

### **2. Methods**

Hardware base of the sanogenetic monitoring complex includes three major appliances adapted to non-invasive screening survey:


7. The method of documentation of pathological consequences rules out the possibility of early prophylactic protection of the population contacting with hazardous industries, which is a priori more efficient and less costly than treatment of the realized risks. By the wide range of effects, all existing real dangers of radiation, physical, chemical, and biological nature can be divided into two categories: 1) risks in the deterministic range of doses and concentrations (doses and concentrations far surpassing than the established thresholds), 2) risks in a stochastic range of doses and concentrations (doses and concentration near the established thresholds). In the deterministic range of doses and concentrations, the biological effects strictly depend on doses and concentrations of anthropogenic factors and can be detected by existing methods of epidemiological analysis. In the stochastic range of doses and concentrations of anthropogenic factors, the consequences strictly depend on individual

In the human body, many regulatory systems operating at different levels of organization provide sensitivity or resistance to both external and internal factors. On the results of analysis of functional adequacy of sanogenesis systems predicts the level of resistance or sensitivity to relatively tolerable doses and concentrations of anthropogenic influences. Hence, sanogenetic monitoring is more informative for the range of doses and

Due to individual variability of functioning of sanogenetic processes, the same anthropogenic factor in equal doses and concentrations will cause certain effects in some organisms (sensitive), will not cause in others, and will induce resistance in the third. This implies that at the population level three subpopulations should be determined at relatively low-dose and low-concentration exposures: sensitive, neutral, and super resistant. The ratio between these subpopulations can eventually serve as a criterion of population risk from

Any stable fixation of the pathological trace is preceded by processes of dysregulation of the corresponding functions. The most probable pathological outcomes can be predicted on the basis of the results of polysystemic sanogenetic monitoring by detecting dysregulation in certain systems of the organism (cardiorespiratory, psychomotor, and metabolism systems). Monitoring is carried out using computerized measurement instrumentation and data processing systems, which provides the basis for strict quantitative assessment of the dynamics of risk for the studied populations. The risks assessment goes from the instrument of control to the rank of controlled processes, which is the basis for successful operation of

Hardware base of the sanogenetic monitoring complex includes three major appliances


provide a reliable argument for choosing optimal preventive measures.

sensitivity of biological objects, including humans.

concentrations that cause stochastic effects.

potentially hazardous industries.

adapted to non-invasive screening survey:

transducer, and electrocardiogram;

this exposure.

**2. Methods** 

chemical, radiation, climatic, noise, vibration, and other impacts. Hence, they do not


All tests were performed with strict adherence of general bioethical standards.

Individual *functional sufficiency of the cardiorespiratory system* was evaluated using a Spiroarteriocardiorhythmograph instrument complex (SACR, recommended by Ministry of Health Care and Social Development of the Russian Federation for clinical use; registration certificate #29/03020703/5869-04, St. Petersburg) allowing simultaneous recording of the heart, vascular, and respiratory rhythms. The method makes it possible to calculate the relative contribution of sympathetic and parasympathetic autonomic nervous system (ANS) into heart rate and BP regulation, integrated values of cardiogram intervals, parameters of lung ventilation, baroreflex parameters, etc.

Electrocardiogram (ECG) was recorded in standard lead I over 2 minutes. The timeamplitude parameters of PQRST complex and heart rhythm variability (HRV) were evaluated using statistic, geometric, and spectral parameters. HRV power in different frequency bands determined using Fourier-transform analysis characterizes ANS activity and the function of the central mechanisms of heart rate regulation. Three frequency bands can be distinguished in spectra: very low frequency (VLF, 0-0.04 Hz), low frequency (LF, 0.04-0.15 Hz), and high frequency (HF 0.15-0.4 Hz), which are measured in absolute values of power (msec2). These values can also be presented in standardized units (LFn, HFn) calculated as the ratio of each spectral component to their sum. Index of autonomic balance (AB=LF/HF) and index of centralization (C=(VLF+LF)/HF) were calculated from HRV spectral parameters.

Peripheral systolic and diastolic blood pressure (SBP and DBP, respectively) and their variability were measured on middle phalanx using the method of Penaz. From the parameters of BP pulse wave, hemodynamic parameters, stroke volume, and cardiac output were calculated using phase analysis of cardiac cycle and BP. Spontaneous arterial baroreflex sensitivity (BRS=LFM/LFSBP) was also evaluated. From geometric parameters of HRV (mode, mode amplitude, amplitude of oscillations, etc.), autonomic balance index, parameter of adequacy of regulation processes, autonomic rhythm index, and regulatory system strain index (SI) were calculated.

For evaluation of functional reserves of the cardiovascular system, a functional test with increased "dead space" was used (Trukhanov et al., 2007). Reactivity of the cardiovascular system was evaluated by changes in the parameters describing its function (in %) during ECG recording in spirometric mask in comparison to ECG recorded without the mask. The time of inspiration and expiration, volume rate of inspiration and expiration, and respiratory volume of quiet breathing in an averaged cycle were evaluated. Parameters of forced expiration (vital capacity of the lungs and volume of forced expiration) were also measured.

The study of the *latent period of simple sensorimotor reaction and other psychomotor parameters* was performed using a specific instrument called "CMM" (computer movement meter), Registration Certificate # 29/03041202/5085-03. The accuracy of measuring the time of simple sensorimotor reaction was 1 ms. A subject was placed in a comfortable chair,

Polysystemic Approach to Risk Assessment 189

*Statistical processing* of experimental data was performed using nonparametric tests (Kruskal-Wallis, Mann-Whitney U test, Spearman correlation coefficient, and Fisher exact test), because empirical data did not conform normal distribution according to Kolmogorov-Smirnov test. The differences between the parameters within the group were evaluated using paired Wilcoxon T test. The data are presented as M±SEM. The significance level

Device complex and methodological approaches have been tested during screening

Laser correlation spectroscopy was used for the analysis of blood serum from individuals exposed to repeated or single irradiation. Changes in spectral characteristics associated with shifts in the homeostatic system were revealed (Akleyev, Kisselyov, 2000). Further studies in this field showed that even single radiation exposure leads to metabolic shifts in the organism towards predominance of catabolic processes. Redistribution in the blood serum spectrum was also directed towards accumulation of the low-molecular fraction (Akleyev &

We examined workers (n=328) employed at nuclear fuel cycle plant in Electrostal' town contacting with open (shop #1, uranium and its products, MPC level) and sealed (shop #2) sources of radiation, or exposed to combined influence of radiation (shop #3, uranium and its products, above MPC level), chemical, and other factors. Blood serum samples from

Comparison of the percentage of metabolic shifts in different shops showed that the percentage of normological spectra was low in shop #1; the percentage of anabolic shifts in

% normal catabolis anabolic mixed

control shop 1 shop 2 shop 3

Fig. 1. Incidence of different types of metabolic shifts at nuclear fuel cycle plant of

workers not contacting with radioactive materials were used as the control (n=16).

all shops was decreased in comparison with the control (Fig. 1).

**3.1 Screening examination of workers of the nuclear fuel plant** 

examination of workers of the nuclear fuel plant.

was 5%.

**3. Results** 

Kisselyov, 2000).

Electrostal' town.

while his/her hand was placed on a special handle and lever, which, in turn, may revolve around a vertical axis. The fulcrum of the rotating segment was treated as the upper third of the forearm. In this position the forearm and wrist could commit abduction and adduction. Mechanical resistance to rotational movement of the forearm was insignificant and therefore ignored in the calculations. Initially, the lever with forearm and wrist resting on it was fixed on the zero position by electromagnetic stoppers. Participants were instructed to focus on the cross in the centre of the screen, and to adduct the handle of the lever with their forearm and wrist as quickly as possible in response to the visual signals started at random intervals (4–8 seconds). The visual signals (vivid light) were slightly peripheral to the central visual field, in order to potentially speed up sensor motor reaction. At the instance of switching on light the electromagnetic stoppers were simultaneously removed and the lever was able to move freely. The latent period of the motor response (reaction time – RT) was measured from the moment of switching on light until the angular displacement of the the handle with forearm at 1 degree was recorded by a computer. The subjects were not limited in the amplitude of translation of the lever. The study was performed for both dominant and subdominant hands. Each subject received 16 signals for reaction with each hand. Training before the experiment included reaction for 10 visual stimuli which were randomly distributed in time.

*Subfractional composition of blood serum* was analyzed using laser correlation spectrometer (LCS, certificate of Committee on New Medical Instrumentation, Ministry of Health Care and Social Development of the Russian Federation, RU.C. 39.003.A N 5381, St. Petersburg). The method is based on changes in spectral characteristics of monochromatic coherent helium-neon laser radiation due to light scatter in disperse system (blood serum, urine, and other biological fluids) (Karganov et al., 2011). The degree of this scatter is proportional to particle speed, which depends on its hydrodynamic radius. The spectra of blood serum samples (0.2 ml) were recorded and processed routinely (Karganov et al., 2011b).

*Evaluation of individual sensitivity to ionizing radiation* was carried out on mitogen-stimulated peripheral blood lymphocytes. The cells were cultured in glass flasks in a medium containing sterile embryonic calf serum ("Perbio-HyClone", USA), RPMI-1640 medium with 25 mM HEPES and sodium bicarbonate, phytohemagglutinin ("Sigma", USA), glutamine, and antibiotics.

The cells were irradiated on a "Luch-1" γ-apparatus (Medical Radiological Research Center, Russian Academy of Medical Science, Obninsk) at 0.25 Gy/min radiation power and 65 cm distance. The adapting dose (AdD) was 0.05 Gy and the damaging dose (DD) was 0.5 Gy. Three samples for each examinee were irradiated according to the following schemes: 1) AdD at G0 stage of the cell cycle without DD (for evaluation of the effect of AdD); 2) DD at G2 stage of the cell cycle 48 hours after AdD; 3) DD without AdD (for evaluation of DD). The cells were incubated and fixed using standard methods (Hungerford D.A., 1965). The preparations for routine analysis were stained with azure and eosin and examined under a microscope in transmitted light under oil immersion at ×1000. The following chromosome aberrations were counted: chromatid and isochromatid fragments and symmetrical and asymmetrical chromatid exchanges. For each donor, 100 metaphase plates per term were analyzed. The RAR coefficient at different irradiation doses was calculated by the ratio of the number of chromosome aberrations: RAR=control + DD/(AdD+DD); RAR≥1.5 indicates the presence of the adaptive response.

*Statistical processing* of experimental data was performed using nonparametric tests (Kruskal-Wallis, Mann-Whitney U test, Spearman correlation coefficient, and Fisher exact test), because empirical data did not conform normal distribution according to Kolmogorov-Smirnov test. The differences between the parameters within the group were evaluated using paired Wilcoxon T test. The data are presented as M±SEM. The significance level was 5%.
