Behavioral Toxicodynamics and Kinetic profiling of PCB congeners in Swiss albino mouse dosed with Aroclor 1232 through inhalational route: A pivotal risk assessment study

Behavioral Toxicodynamics and Kinetic profiling of PCB congeners in Swiss albino mouse dosed with Aroclor 1232 through inhalational route: A pivotal risk assessment study | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Behavioral Toxicodynamics and Kinetic profiling of PCB congeners in Swiss albino mouse dosed with Aroclor 1232 through inhalational route: A pivotal risk assessment study Sivaselvakumar Muthusamy, Ramanujam Narayanan This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4715900/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Introduction: Polychlorinated Biphenyls pose hazards to humans as environmental and occupational workplace pollutants, despite which, though they are regulated in India, their use is not yet banned. Hence, the pharmacokinetics and behavioral effects of PCBs after inhalational exposures of Aroclor 1232 in Swiss albino mice were assessed in this study, with an objective to correlate these effects with monoamine levels in brain and extrapolate them to rodent and human risk assessments. Methods: A whole body inhalational chamber has been utilized to expose mice to different doses of Aroclor 1232 for single six-hour durations. The plasma and brain samples were used to estimate the corresponding concentration-time profiles of two representative congeners of PCBs (PCB 77 and PCB 180) and calculate their pharmacokinetic parameters. These variables were then correlated with mouse behavior using Irwin scale, and monoamine levels at Tmax. Results & Discussion: A definite dose-dependence of PCB 77 and PCB 180 kinetic parameters was identified in plasma and brain. PCB 77 exhibited more accumulation in brain than plasma, whereas PCB 180 accumulated more in plasma than brain. Mood, autonomic scores, and miscellaneous sub domains were significantly different from control group scores in these domains. The changes in the mood and miscellaneous domains on the Irwin scale could be ascribed to higher brain PCB 77; whereas changes in autonomic functions could be ascribed to higher plasma PCB 180 levels. Conclusion: The pharmacokinetic parameters of PCB 180 and PCB 77 after acute inhalational exposures to Aroclor 1232 were obtained and this was temporally correlated with disruption of behavioral scores and monoamine levels in dose-dependent manner. Polychlorinated Biphenyl Aroclor 1232 Inhalation toxicity Behavior Irwin scale Monoamines Mood Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Poly-Chlorinated Biphenyls (PCBs) are a chemical class of 209 congeners that are known to bio-accumulate, bio-magnify, bio-concentrate and persistacross all the levels of the food chain. Though the production, distribution, and supply of their commercial mixtures like Aroclors and Kaneclors have beenstrictly regulated in most of the member countries by the United Nations Stockholm Convention [ 1 ] their replacement by best alternatives and their properdisposal are not practically feasible in many regions regulated. This is due to their favourable thermal inertness, hydraulic and dielectric nature, because ofwhich their use in electronics, rubber and plastic industries are unavoidable; additionally, higher lipophilicity and halogenated nature make theirenvironmental elimination practically impossible [ 2 ]. The 209 chemical congeners of PCBs can be further classified based on the number of chlorine substitutions, as low chlorine (1–4 chlorinated biphenyls)and high chlorine (5–10 chlorinated biphenyls) homolog families, each family having one to many congeners differing in the specific spatial chlorinesubstitution on the biphenyl nucleus. The number and the exact site of the chlorine substitution determine the nature of the congener’s structure-activityrelationship and its metabolism in-vivo, which determine its pharmacokinetics, pharmacodynamics, cumulation, and toxicological profiles. For example,ortho-substituted PCBs are known to be highly neurotoxic [ 3 ], whereas co-planar PCBs are known to possess cardiometabolic toxicities [ 4 ]. Similarly, DioxinlikePCBs are proved to have potential to bind to Aryl-Hydrocarbon receptors in the cell nucleus and cause neuroendocrine disruption [ 5 ]. Highly chlorinatedPCB congeners are known to be highly lipophilic and persist in the environment, with long biological half-lives [ 6 ]. However, the complex individualchemistry of these congeners and the variable molar composition of their restricted commercial mixtures make the predictability of the biologicalpharmacokinetics of PCB congeners after dosing of these mixtures complicated and difficult in-vivo. Aroclor 1232 is a semi-volatile commercial PCBmixture that was selected as the test substance for its ability to cause occupational air pollution indoor and outdoor. It is low chlorinated (32% Chlorine bymolar mass) –hence, may not have long environmental half-lives despite in-vivo accumulation. It also has low vapor pressure such that it can also leachsoil from air and be carried out to water bodies [ 2 ]. PCB 77 is a Dioxin like tetra-chloro-biphenyl (Tetra CB) that is also semi-volatile but is highly potent incausing neuroendocrine toxicities after binding to Aryl Hydrocarbon receptors [ 4 ]. PCB 180 is a highly lipophilic hepta-CB congener that is known to persistin biological systems and cause cardio-metabolic toxicities [ 6 ]. PCB 180 has higher octanol-water partition coefficient than PCB 77 [ 2 ]. As a blind screening tool, the Irwin neurobehavioral scale is a validated scale to be used to assess the cross sectional behavioral, autonomic, neurological,and physiological status of the laboratory mouse, both in normal and treated animals [ 7 ]. Additionally, it can be used to delineate the pharmacologicaleffects of diverse groups of drugs acting on the nervous system. Using existing HPLC-FLD-based approaches for measurement of monoamines, the studyevaluated the behavioral changes with pharmacokinetic time-points in relation to monoamine concentration changes. This study aims to predict the pharmacokinetic variables of PCB 77 and PCB 180 after inhalation dosing of a PCB commercial mixture, viz., Aroclor 1232, in mice housed in a whole-body inhalation chamber, in an acute exposure protocol. The study then enables to study further the relationships of the PCB 77and PCB 180 levels in brain and plasma with behavior scores assessed by Irwin scale. After dosing this mixture in mice via inhalation chamber, thebehavioral effects of PCB 77 and PCB 180 have been determined using estimations of their plasma and brain concentration-ratios at geometricallyprogressive dose increments. Materials and Methods Chemicals : Aroclor 1232 for inhalation exposure was purchased from Sigma-Aldrich Inc., India. Reference standards PCB 77, PCB 180, PCB 101, 5-Hydroxytryptamine (5-HT), Dopamine (DA), Nor-adrenaline (NA) and Paracetamol were obtained from Sigma-Aldrich Inc., India. PCB 101 was used as an Internal Standard (IS). Solvents including methanol, tert-Butyl Methyl Ether (TBME) and acetonitrile (ACN) were purchased from Sigma-Aldrich Inc., India. High-purity water for chromatographic analyses was purchased from Merck-Millipore Inc., India. Animals: Institutional Animal Ethics Committee (IAEC) approval was obtained prior to the conduct of the study for utilization of Swiss albino mice, which has been recognized by the Indian Regulatory authority,the Committee for the Purpose of Control and Supervision of Experiments on Animals [8]. All mice were Females about 16-32 gm at initiation and were housed in pairs per cage. Before study initiation, all the animals were acclimatized, and allocated to the three dose levels randomly. Except during the exposure period, they were provided with water and feed ad-libitum and housed under standard circadian rhythm and housing conditions as per OECD guidance for using humane endpoints in toxicity studies as applied to inhalation toxicity [9]. Methods: Inhalation Chamber and the Exposure protocol Components of a whole-body inhalation chamber which was used for exposure of test chemical in mice consisted of five units – the inlet, the animal exposure-control unit, the outlet, the connecting lines, and the external indoor environment [10]. Exposures to Aroclor 1232 consisted of three test doses in geometric progression (2.0, 4.0 and 16.0 mg.m-3). Each dose of the test substance was given to a group of 36 mice in the inhalation chamber as per OECD guidelines for sub-acute inhalation toxicity [9]. A cohort of four animals per time point was procured out from the chamber for each time point (pre-dose, 2.0, 4.0, 6.0, 8.0, 12.0, 24.0, 36.0, 48.0 hours) for a total of nine time-points per dose level. The internal chamber environmental variables like oxygen content, relative humidity, temperature, pressure, and flow rate of air through the inlet along with the Aroclor 1232 solution were monitored and controlled for ensuring proper animal welfare. Irwin Behavior assessment It required about three to eight minutes of testing per mouse for its behavioral rating scores using Irwin scale. The scale consists of about forty-seven different functions and processes, divided into eleven domains, each affecting the central, peripheral, and somatic, and the autonomic nervous system, with effects related to cardiovascular and respiratory systems. Certain of these functions or sub domains can be tested by observation, whereas others can be assessed in the mouse after gentle maneuvers. Each sub domain has a minimum score, a normative score, and a maximum score, ranging from 0-4. The initial undisturbed observation consisted of assessment of body position, locomotor activity, exophthalmos, respiration, tremors, twitches, convulsions, exploratory activity, hostility, and social conglomeration, as the mice remained inside the chamber. The animal was scored on a scale of 0-4 for each of these sub domain functions. After completion of this initial observation, the mouse was transferred from the chamber into the cage and then gently manipulated towards the floor of the chamber for response grading to assess the transfer arousal, spatial locomotion, palpebral closure, startle response, piloerection, tremor grading, gait and limb rotation. Following these scoring steps, the animal is lifted vertically by its mid tail above a wired mesh grid, and lowered slowly for assessment of visual placing response, i.e., extension of all limbs before contact. Grip strength and positional struggle behavior were then assessed. Body tone, gross signs of hypothermia, pinna and corneal reflexes, toe-pinch response were then graded. After this, the mouse was transferred to the chamber and graded for alley progression, withdrawal responses, touch-escape response, and positional passivity. Finally, autonomic signs like diarrhea, skin / fur colour, limb tone, abdominal tone, pupillary reflex, lacrimation, salivation, pupillary size, and provoked biting response (for aggression) were observed followed by tail-pinch response, righting reflex, provoked freezing, grasp irritability, vocalization and defecation and urination assessments [7]. Sample collection Methods of Euthanasia: Each animal was euthanized with Ketamine (55-77 mg/kg intraperitoneally) at the sacrifice as per AVMA Guidelines. Methods of Anesthesia / Analgesia and Humane endpoints for animal welfare: All mice were observed for vital parameters like Heart rate, Respiratory rate and involuntary movements after ketamine overdose administration. If tachycardia (HR >2 beats / s) / tachypnoea (RR > 5 /10 sec) / involuntary movements (Flexion / extension of limbs) were present even after Ketamine overdose, the animal was given Xylazine 4 mg/kg overdose I.P. immediately to avoid suffering. As per Refinement (Russel & Burch’s 3Rs) this step alleviates animal suffering and conforms to humane endpoints of animal welfare. Anesthesia & analgesia were not used for the study as they are not compatible with the study protocol. Death was immediate with Xylazine overdose (within next 2 minutes). Specimen / sample coding : All animal samples were collected in coded sample tubes as plasma or brain in 1x PBS (ice cold). The codes were later decoded only upon statistical analysis. Plasma collection and processing: Before euthanizing PCB-exposed mice at each time point for dissection and homogenization of brain samples by high dose Ketamine (55 mg/kg I.P.) (Leary et al., 2013), their blood samples were collected by a terminal cardiac puncture (maximum 0.2 ml) in tubes premixed with 0.012 M buffered calcium disodium EDTA in the ratio of 3:1 of anticoagulant solution: blood, and these were then centrifuged at 3000 rpm for 8 minutes at 4°C immediately. The centrifuged sample volumes were then separated for their plasma component, and the plasma was stored at -80 ± 10°C, until analysis. Brain collection and processing: The euthanized mice brains were dissected out as whole; homogenized for storage in 1% of ice-cold Phosphate Buffered Saline (PBS) solution in -80 ± 10ºC until analysis. Blank brain samples were collected at 0 hours pre-dosing for calibration purposes. Each brain sample was divided into two equal portions for estimation of PCB levels and monoamine levels, respectively. Instrumentation and Chromatography conditions for PCB estimation: The Ultra-High-Performance Liquid Chromatography (UPLC) instrument consisted of a Waters Acquity H class UPLC system equipped with a quaternarypump and 96-vial autosampler coupled with a diode array UV detector (Waters, Milford, MA, USA). The chromatographic separation was performed on anAcquity UPLC BEH C18 column from Waters (2.1 mm × 100mm; 1.7μm). The column temperature was set at 35ºC and the autosampler was kept at 15ºC. The mobile phase composed of a mixture of acetonitrile (85%, v/v) and water (15%, v/v) at a flow rate of 0.3ml/min. Before analysis, the mobile phase wasfiltered through a 0.22μm membrane filter and degassed by ultra-sonication. A 5μl injection of each sample was loaded on to the system and total analysistime was 6 min. The Diode Array Detector (DAD) was set at 215 nm. The sampling needle was washed with 300μl of strong wash (ACN/water, 65/35) toreduce carry-over and 300μl of weak wash (ACN/water, 30/70). Data acquisition was done using Empower 3 software version 1.0 (Waters, USA) [11]. Parent monoamine analysis measurements: 5-HT, DA and NA stock solutions of 1mg/mL were prepared by dissolving a suitable amount of single standard in acetonitrile. Mixture of stock solutions 5-HT, DA and NA (100 μg/mL) were prepared in acetonitrile. The High-Performance Liquid Chromatography (HPLC) instrument consisted of a Waters 515HPLC system equipped with an isocratic pump and coupled with a fluorescence detector (Waters, Milford, MA, USA). The chromatographic separation wasperformed on a Gemini-C18 column from Phenomenex (4.3 mm × 50mm; 3μm).Buffer was prepared using 70 mM potassium dihydrogen phosphate, 0.1mM dipotassium EDTA salt, 1.1 mM heptane sulfonic acid and 0.4 % Trifluoracetic acid added to 1000 mL water and adjusted to a pH of 3.14 usingsaturated citric acid solution. The mobile phase composed of a mixture of methanol (10%, v/v) and buffer (90%, v/v) at a flow rate of 0.5ml/min.Paracetamol used as internal standard (IS). The mobile phase was filtered through a 0.22μm membrane filter and degassed by ultra-sonication. Thefluorescence detector was set excitation of 285 nm and emission at 345 nm respectively. Data acquisition was done using Empower 2 software version 1.0(Waters, USA) [12]. Statistical analysis: Data were represented as mean ± standard deviation. Win-Non-Lin software 5.1 was used to calculate the pharmacokinetic parameters by noncompartmentalmodel (Pharsight Corporation, CA, and USA). Pearson correlation was used to assess the relationships between behavioral scores in eachsub-domain and the time-points for each dose level. One-way Repeated Measures ANOVA was used to evaluate the significance of each correlationcoefficient obtained pair wise in Pearson correlation test. Hill model was used to model dose-response-effects versus brain : plasma ratios for each congenerestimated using Inhibitory-Dose (ID-50) modeling. Pearson Correlation, one-way Repeated Measures Analysis of Variance (RM-ANOVA) and Dose responsemodeling (Hill model, and monoamine analysis) were performed using GraphPad Prism version 8.4.2 for Windows, GraphPad Software, San Diego,California USA, www.graphpad.com. Results The pharmacokinetics of PCB 77 and PCB 180 in plasma and brain were similar with small but notable differences in a dose-dependent manner(Supplemental files 1A and 1B). These parameters could be correlated to central behavioral effects based on brain pharmacokinetics, and autonomicnervous system effects based on plasma pharmacokinetics, in dose-dependent and time-dependent manner (Supplemental files 3A and 3B). Generally,brain accumulation of PCB 77 was more significantly responsible for behavioral disruption than PCB 180 accumulation in plasma – based on brain-plasmaratios of both these congeners (Supplemental files 2A and 2B). Pharmacokinetics profile of PCB 77 at 2.0, 4.0 and 16.0 mg.m -3 in mice plasma :The mean maximum concentration attained, Cmax of PCB 77 in plasma was 88.32 ± 8.28 ng.ml -1 for the 2 mg.m -3 group. The time taken to achieve this value of Cmax in the plasma of low dose exposure group was 3.50 ± 1.00 hr. The AUC 0-t for PCB 77 in plasma was 1321.19 ± 55.72 ng.hr.ml -1. The Area Under the plasma Concentration-time (infinity) Curve, AUC 0-∞ in plasma was 1626.02 ± 64.13 ng.hr.ml -1 for PCB 77 after dosing 2 mg.m -3 of Aroclor 1232. The elimination rate constant, K el in plasma for PCB 77 after dosing mice with 2 mg.m -3 of Aroclor 1232 by inhalation route was 0.03 hr-1. Plasma half-life of PCB 77 in this exposure group was 20.17 ± 1.38 hr. The Mean plasma Residence Times (MRT) for PCB 77 in this dose level was 16.51 ± 0.42 hr. The plasma apparent volume of distribution (Vd) and clearance (CL) for PCB 77 after dosing Aroclor 1232 in this group of mice were 35858.25 ± 3073.55 and, 1231.46 ± 49.43 ml.hr -1.m-2 , respectively. In the 4 mg.m-3 exposure group, the Cmax and Tmax of PCB 77 in plasma were 245.62 ± 39.58 ng.ml -1 and 4.00 hr, respectively. The AUC 0-t and AUC 0-∞ of PCB 77 in this intermediate dosing group were 2749.28 ± 392.00 and 2957.28 ± 413.20 ng.hr.ml-1 in plasma. The Kel values were 0.06 hr-1 in plasma of themice treated with 4 mg.m -3 group, while the half-life was 12.72 ± 0.36 hr in this group. The MRT in the intermediate dose exposure groups was 14.31 ± 0.49hr in the plasma. Finally, the Vd and CL in plasma of this exposure level were 25209.01 ± 3835.06 ml.m -2 and 1375.07 ± 215.15 ml.hr -1.m-2 .In the 16 mg.m -3 group, the Cmax of PCB 77 was 727.40 ± 42.52 ng.ml-1 in plasma and Tmax was 4.00 hr. The mean AUC 0-t and AUC0-∞ in the plasma of thishigh dose group were 7565.21 ± 148.44 and 8013.39 ± 263.11 ng.hr.ml -1 . In the high dose group, the K el and t-1/2 in plasma were 0.06 hr - 1, and 11.17 ± 1.54hr, respectively. The MRT of PCB 77 in plasma of the mice treated with high dose of Aroclor 1232was 14.29 ± 0.24 hr. The high dose group Vd and CL in plasma was 32099.78 ± 3593.04 ml.m-2 and 1998.28 ± 65.96 ml.hr -1 .m -2 in plasma.The mean plasma concentration-time profile of PCB 77 in mouse plasma after inhalation of PCB mixture at three different dose levels are presented inFigure 1, inset A, and Table 1. Pharmacokinetics profile of PCB 77 at 2.0, 4.0 and 16.0 mg.m-3 in mice brain :The mean maximum concentration attained; Cmax of PCB 77 was 69.43 ± 10.54 ng.ml -1 in brains of mice treated with 2 mg.m -3 of Aroclor 1232 via inhalation route. The Tmax of PCB 77 in brain tissues of mice exposed to this dose was 3.50 ± 1.00 hr. The AUC0-t for PCB 77 in brain was 1254.97 ± 57.51ng.hr.ml -1 . The AUC0-∞ was 1469.62 ± 161.80 ng.hr.ml -1 for PCB 77. The elimination rate constant, Kel in brain for PCB 77 was 0.05 hr-1. Brain half-life ofPCB 77 in this exposure group was 16.20 ± 4.36 hr. The Mean brain Residence Times (MRT) for PCB 77 in this dose level was 17.07 ± 1.93 hr. The valuesfor the apparent volume of distribution and tissue clearance of PCB 77 in brain were 31481.64 ± 5927.66 ml.m -2 respectively, and 1373.32 ± 150.80 ml.hr -1 .m - 2, respectively.In the 4 mg.m -3 exposure group, the Cmax and Tmax of PCB 77 in brain were 175.13 ± 6.42 ng.ml -1 , and 4.00 hr, respectively. The AUC 0-t and AUC 0-∞ of PCB77 in this group were 2581.12 ± 136.46 and 2366.40 ± 142.19 ng.hr.ml-1, respectively. The K el values were 0.05 hr-1 in brain of the mice treated with 4 mg.m -3 group, while the half-lifeof PCB 77 in brain was 14.20 ± 0.58 hr in brain. The MRT in the intermediate dose exposure group was 15.05 ± 0.32 hr in brain, while the Vd and CL in brain of this exposure group were 31856.13 ± 3012. 73 ml.m-2 and 1553.12 ± 85.97 ml.hr -1 .m -2 .In the 16 mg.m -3 group, the Cmax of PCB 77 was 526.21 ± 28.63 ng.ml -1 in brain and Tmax was 4.00 hr. The mean AUC 0-t and AUC 0-∞ in the brain of thisgroup were 6798.93 ± 340.47 ng.hr.ml -1 and 7100.56 ± 413.77ng.hr.ml -1. The K el and t-1/2 in brain were 0.07 hr-1, and 10.82 ± 1.29 hr, respectively. The MRTwas 14.39 ± 0.30 hr. The Vd and CL in brain were 35070.00 ± 2110.32 ml.m-2 and 2258.90 ± 127.06 ml.hr -1 .m -2 .The mean brain concentration-time profile of PCB 77 in mouse brain after inhalation of PCB mixture at three different dose levels are presented in Figure 1,inset B, and Table 1. Pharmacokinetics profile of PCB 180 at 2.0, 4.0 and 16.0 mg.m -3 in mice plasma : The mean Cmax of PCB 180 in plasma after 2 mg.m -3 inhalation of Aroclor 1232 was 66.49 ± 5.22 ng.ml -1 . The time taken to reach this value of Cmax in plasma was 2.00 hr. The AUC0-t and AUC0-∞ attained by PCB 180 in plasma in this dose level were 763.27 ± 38.80 and 1226.52 ± 136.72 ng.hr.ml -1 ,respectively. The Elimination rate constant of PCB 180 for its elimination from plasma at this dose level was 0.02 hr -1 . Its half-life in plasma was 28.95 ±3.39 hr. Its MRT in plasma was 13.11 ± 0.96 hr at this dose level. The plasma Vd and CL of PCB 180 after administration of 2 mg.m -3 of Aroclor 1232 for 6hr were 68083.41 ± 1151.95 ml.m -2 , and 1646.03 ± 184.36 ml.hr - 1.m -2 .The mean Cmax of PCB 180 in plasma in the 4 mg.m -3 group was 370.21 ± 42.59 ng.ml -1 , and the time taken to reach this concentration in plasma (Tmax) was 2.00 hr. The AUC 0-t and AUC 0-∞ were 4369.75 ± 730.55 and 5425.39 ± 408.95 ng.hr.ml -1 . This congener’s elimination rate constant and half-life were0.04 ± 0.01 hr -1 and 20.04 ± 6.43 hr, respectively. MRT in plasma for this group was 15.43 ± 0.85 hr; with a Vd and plasma CL of 21729.82 ± 8734.97 ml.m -2 and 740.51 ± 57.37 ml.hr -1 .m-2, both somewhat less compared to the lower dose level.In the 16 mg.m -3 exposure, the time taken to reach a plasma Cmax of 829.57 ± 31.40 ng.ml -1 was 6.00 hr. The corresponding AUC 0-t and AUC 0-∞ for theplasma-concentration-time relationship in this group were 13192.39 ± 498.68 and 15636.03 ± 1053.10 ng.hr.ml -1 . The K el and t-1/2 in this group were 0.04 ±0.01 hr-1 and 18.21 ± 1.94 hr, respectively, and the MRT was 16.24 ± 0.32 hr. The Vd and CL of PCB 180 from plasma at this dosing level were 26825.47 ± 1287.33 ml.m -2 and 1026.94 ± 72.53 ml.hr -1 .m -2 , respectively.The mean plasma concentration-time profile of PCB 180 in mouse plasma after inhalation of PCB mixture at three different dose levels are presented inFigure 2, inset A, and Table 2. Pharmacokinetics profile of PCB 180 at 2.0, 4.0 and 16.0 mg.m -3 in mice brain: The peak concentration of PCB 180 attained in the brain in the 2, 4 and 16 mg.m -3 groups were42.04 ± 4.83, 135.49 ± 4.95 and 429.35 ± 14.37 ng.ml -1 ,respectively, showing a geometrically progressive trend with dose. The Tmax of this congener in brain were 2.0 ± 0.0, 2.0 ± 0.0, and 6.0 ± 0.0 hr in the threedose levels, respectively. The AUC0-t in brain were 631.12 ± 46.60, 2449.37 ± 186.79, and 8332.99 123.54, ng.hr.ml -1 respectively with dose increments; theAUC 0-∞ for PCB 180 across the three dose levels were 915.48 ± 106.63, 3896.91 ± 367.99, and 10984.01 ± 590.37 ng.hr.ml -1 , respectively. The K el in brain were 0.03, 0.02 and 0.03 hr -1 across the three graded doses of Aroclor 1232. The half-lives of PCB 180 elimination from brain were 22.17 ± 3.24, 31.43 ±3.10, and 22.73 ± 3.57 hr, while the MRT were 13.88 ± 0.21, 19.45 ± 0.64, and 18.72 ± 0.21 hr correspondingly, in the brain. The mean apparent Vd of PCB180 in the mouse brains in the 2, 4 and 16 mg.m - 3 doses were 69838.36 ± 5306.26, 46934.13 ± 6867.21 and, 47593.68 ± 4968.74 ml.m -2 and its clearancewere 2207.04 ± 256.46, 1033.24 ± 96.14 and 1459.71 ± 75.49 ml.hr -1 .m -2 .The mean brain concentration-time profile of PCB 180 in mouse brain after inhalation of PCB mixture at three different dose levels are presented in Figure 2,inset B, and Table 2 (Supplemental files 1A and 1B). Dose dependence of plasma PCB 180 and PCB 77 : Aroclor 1232 was dosed to Swiss albino mice at three dose levels – 2.0 (low dose), 4.0 (intermediate dose) and 16.0 (high dose) mg.m -3 . PCB 77Cmax in plasma was eight times higher in high dose compared to low dose level. However, this was only three times higher in intermediate dose group compared to 2.0 mg.m -3 dose level and three times higher for high dose compared to 4.0 mg.m -3 . In plasma, PCB 180 Cmax was six times and twelve timeshigher for intermediate and high dose groups compared to 2.0 mg.m -3 dose levels. The high dose level Cmax for plasma PCB 180 was twice thanintermediate dose level. Dose-dependent translation of brain to plasma PCB 77, 180 ratios: PCB 77 brain to plasma ratios were similar for intermediate and high dose level, which reached about 1.1 times for lower dose level. The ratios for PCB 180 were three times in the intermediate dose level compared to high dose; the ratio was 5 times higher in low dose compared to the high dose level, and 1.7times high in intermediate dose compared to low dose (Figure 3, Table 3, Supplemental files 2A and 2B). Effect of brain to plasma ratio of PCB 77 and PCB 180 in Swiss albino mice: Hill model was used to quantify the Inhibitory Dose, ID-50 values for PCB 180 and PCB 77 to depress behavioral scores at the same level of brain toplasma ratios (Figure 3, and, 4). PCB 180 ID-50 was 7.645 mg.m -3 , and this was higher than PCB 77 ID-50 values (7.382 mg.m -3 ). The slopes of lines were13.89 and 12.42, and Y-Intercepts for behavioral disruption were 38.23 and 37.16 respectively for PCB 77 and PCB 180. Effects of Aroclor 1232 on behavior : There was a significant difference in total scores at 3 and 4 hours for all three doses. Significant difference in mean scores was also observed at 5 hours of single-6-hour exposures for 2 and 4 mg.m -3 doses (p< 0.05), when compared to control group. The sub scores that contributed to greater than 50%deviation from the normative values, as assessed by Mann Whitney U test were distributed across three main domains – mood, autonomic activity, andmiscellaneous functions, for all dose levels. The Cmax of 2, 4, and 16 mg.m -3 dose groups were achieved at 3.5 ± 1.00, 4.0 ± 0 and 4.0 ± 0 hr for PCB 77 inplasma and brain. The Tmax of PCB 180 plasma and brain at 2, 4, and 16 mg.m - 3 dose levels were 2.0 ± 0, 2.0 ± 0 and 6.0 ± 0 hr. A more detailed analysis ofsubdomain score across doses revealed that grooming and restlessness (mood), urination, heart and respiratory rates, skin colour and palpebral opening(autonomic), and exploratory and group clustering behaviors (miscellaneous domain) were contributing to the statistical significance in the behavioralchanges compared to control group (Supplemental files 3A and 3B). Brain monoamine concentrations at Tmaxof PCB congeners after exposures to different doses of Aroclor 1232 : The brain concentrations of Noradrenaline at 4 hr (Cmax of PCB 77 in brain) were 3.8, 1.7 and 2.0 ng/ml across 2, 4, and 16 mg.m -3 doses. Dopamine (DA)reached 4.8, 1.6 and 7.5 ng/ml in 2, 4 and 16 mg.m -3 doses at 4 hr. (Tmax of PCBs in brain).At 4 hr corresponding to Tmax of PCBs in brain, the brain 5-HTlevels were 3.0, 2.6 and 4.3 ng.ml -1 in 2, 4 and 16 mg.m - 3doses. Brain levels of Noradrenaline and Dopamine were reduced as PCB 77 concentrationsreached maximum in brain. PCB 180 levels did not correlate with either central behavioral disruption or brain levels of all three monoamines. Effects of Aroclor 1232 on brain monoamine levels: Relationship with behavioral scores: Brain monoamine levels were estimated using HPLC-FLD in the first six hours for each dose level. Brain monoamine levels followed a linearrelationship for 5-HT, exponential relationship for NA and non-monotonic or bimodular for DA in the first six hours of exposures. These could be correlatedwith behavioral scores at Cmax time points for each dose level (Figure 5, Table 4, Table 5). Generally, a decline in mood was noted due to dipping of scores for grooming and restlessness sub-domains. The mean grooming scores across all days dipped at 5 hours for higher dose levels as compared to 4 and 6 hours for 2 mg.m -3 dose. The trough in mean restlessness scores across all days was 3 hours for low dose and this shifted by one hour for each geometric increase in dose (Supplemental files 4A, 4B and 4C). In 2 mg.m -3 exposed animals, the mood was generally depressed due to reduction in grooming at 3 and 4 hours, and increase in restlessness at 3, 4, and 5-hour time points. Grooming scores was back to control values at 5 hours in this dose, whereas increased restlessness did not recover even at 6 hours in this group. In the 4 mg/cu.m dose groups, grooming scores dipped at 3, 4, and 5-hour time points. But restlessness scores increased at 3, 4 and 5-hour time points. In the 16 mg.m-3dose level, the grooming reduction was maximum at 3, 4, and 5 hr after exposure and did not recover even at 6 hr in this dose level. Restlessness was increased 3 hr, after which it came to normal at 4 hr time point in this dose level. At 5 hr after exposure, the restlessness scores increased, which did not recover back in this dose level. In this study, the mean palpebral opening was below the normative value at all time points, with the maximum dip approached at 4 hours for 2 mg.m -3 dose and at 5 hours for higher doses. Although the heart rate was below the normative score during all time points, there was a tendency for maximum bradycardia at 3-5 hours across all dose levels. Similarly, the respiratory rate tended to be below the normative always and to initially dip at about 2-3 hours and then recover to baseline value at 5 hours across all days and dose levels. Urination scores increased at 5 hours for 2 and 4 mg.m -3 doses across the day averages, whereas it reduced at 4 and 5 hours for the 16 mg.m -3 dose. Among the miscellaneous domains, group clustering (social behavior) activity increased with duration of exposure as the animals tended to reduce the surface area of their fur to avoid exposure to the potentially irritating PCBs. This was inversely proportional to the exploratory activity of mice in the chamber. Exploratory activity increased at 3, 4, and 5 hr in 2, 4, and 16 mg.m -3 dose levels. Social behavior increased at 3, 4, and 5 hr in 2 and 16 mg.m -3 dose levels. However, in the 4 mg.m -3 dose level, the social behavior scores were normal at 3 hr, then it decreased at 4 hr, after which it again recovered back to normal at 5 hr (Table 5). Relationship of sub-domain behavior scores across doses: The total behavioral subdomain scores across each time point compared pair wise during single 6-hour exposure to Aroclor 1232 exhibited dose dependence at 2.0, 4.0 and 16.0 mg.m -3 dose levels (F ratio = 18.41, p < 0.0001). The Pearson Correlation coefficients were close to 1.0 for most of the pairwise comparisons. One-way Repeated Measures ANOVA of each correlation coefficients showed that the significance was due to inter-dose variation than inter-individual variability (p = 0.0077). Dose dependence of sub-domain scores at Tmax : Likewise, the behavior scores were statistically significant at Tmax of PCB 77 and PCB 180 (3-5 hours) with p value 0.0011. Friedman test of the correlation coefficients at Tmax was significant across doses (Friedman statistic = 31.04). There was more of behavioral depression than stimulatoryeffects of PCBs at Tmax. Contribution of sub-domains to dose-dependent behavioral effects of PCBs at Tmax: Mood, which composed of five sub-domains namely grooming, restlessness, vocalization, irritability, and fearfulness, was generally depressedacross all doses of exposures. This was generally ascribed to depression in grooming behavior on Irwin scale across all three doses, although restlessnessseemed to increase dose-dependently. Concentrations of PCB 77 and PCB 180 in lung, brain and plasma significantly correlated negatively with mooddepression at 6 hours in each dose level (-0.75 <Pearson ‘r’< -0.85, Figure 6). Grooming depression was maximum at 4 hours and Restlessness wasmaximally increased at 4 hours across all doses of exposure (Figure 7). Of the eleven autonomic system functions including palpebral opening, writhing response, pupil size, exophthalmos, pupil size, urination, salivation, hypothermia, skin colour, heart rate and respiratory rate, palpebral opening, urination and skin colour increased at 5 hours of exposure, especially at 4.0 and16.0 mg.m -3 doses. However, heart rate and respiration, reduced at 3 hours maximally across all three doses. Out of the three miscellaneous domains (social aggregation, hostility, and exploratory activity), social conglomeration increased at 3-5 hours acrossall three doses, and exploratory activities increased at 3-5 hours across 2.0 and 16.0 mg.m -3 dosing groups.Generally, the peak of central behavioral disruption followed the Tmax of PCB 77, whereas PCB 180 Cmax contributed more to the peripheral effects onbehavior (Table 4, and, Table 5, Supplemental files 1A, 1B, and 5). Discussion The Inhalation dosing of mice with Aroclor 1232 in the whole-body inhalation chamber was successful in producing significant plasma and brainconcentrations of the two representative PCB congeners. The pharmacokinetic parameters of these two congeners in the two matrices were hence derivedwith high degree of correlation across the dose levels. Plasma peak concentrations of both congeners across all the three doses were generally one-andhalftimes greater than the brain levels. The time to peak absorption of both the congeners were nearly two times longer for PCB 77 than PCB 180, and this was generally independent of dose and tissue matrix. Higher (nearly twice) AUCs were obtained for PCB 77 in both the matrices than PCB 180 generally across the three dose levels. The plasma AUC was generally higher than the brain AUC for both the congeners across the three dose levels, with dose proportionality. PCBs are known to possess high orders of K ow providing them with high lipophilicity and a capacitance to traverse across biological membranes [ 7 ]. PCBs can travel across biological barriers by both passive diffusion and active transport mechanisms. Hence the individual PCB congeners would be expected to cumulate and have longer biological half-lives. However, as is apparent from the acute inhalation study of Aroclor 1232, dosing PCB mixtures can impart only intermediately long half-lives (in hours) and MRTs for certain key toxic congeners like PCB 77 and even the hepta-CB, PCB 180, in plasma and brain tissue as well. This may also explain the short elimination rate constants, which are inversely correlated with log K ow , and hence lipophilicity of congeners [ 13 ]. Though tetra-CB congeners like PCB 77 do not accumulate in aquatic fishes and other environmental reservoirs [ 14 ], certain high chlorinated congeners can attain significantly toxic concentrations and large AUCs in mouse and human matrices, as can be deduced from the large peak plasma and brain concentrations of PCB 180 and PCB 77, especially in higher doses. Such results have similarly been obtained with other investigations [ 15 ] where PCB 77 concentrations in different areas of the brain like the striatal cortex have been generically less compared to PCB 180 concentrations, after feeding rats with Aroclor 1254 in drinking water and chow. Although a dynamic equilibrium exists among the final tissue concentrations of homologs of PCBs as suggested by studies elsewhere [ 16 ] the concentrations of certain toxic congeners can significantly vary in same organism in different tissues [ 17 ]. In our study, both congeners generally have large volumes of distribution and clearance rates from both plasma and brain, which are dose-dependent and not time-of-exposure-dependent. Many PCBs especially higher chlorinated PCBs like PCB 180 can have active metabolites in plasma and other tissues like methoxy, sulphated and hydroxyl metabolites, which could shorten the elimination rate constants of such congeners in-vivo [ 18 ]. Hence, a change in rate of elimination of a congener from a tissue matrix with increments in dose usually depends on two factors – the rate of metabolism to active metabolites, and the rate of excretion versus distribution constants. Since PCB 180 is more lipophilic chlorinated and may have a greater number of potent active metabolites, its volumes of distribution and elimination half-lives change more drastically with dose increments, than PCB 77. Finally, for most of the pharmacokinetic parameters calculated for both congeners in this study, the variations imparted by the inhalation route is less, and the dose-dependent nature of their linear pharmacokinetics in brain and plasma is of a geometrically graded nature. Since PCB 77 is more of a dioxin like PCB with ability to bind to AhRs and cause neuroendocrine disruption more than PCB 180, this study confirms the hypothesis that environmental reminisce of such PCBs, though seemingly low chlorinated can lead to monoamine and other neurotransmitter alterations in the brain even in rodents, via entry into the biological systems by inhalation routes. Further, such alterations could theoretically lead to neurobehavioral disruptions and neurological disorders if they accumulate in the brain as life progresses. Lipophilicity of PCB congeners depends on number of chlorine atoms on the biphenyl nucleus and absorption of PCBs onto biological membranes in turn depends on lipophilicity and steric hindrance caused by position of chlorine on biphenyl congener [ 19 , 20 ]. Ultimately absorption of PCBs in biological systems is influenced by the physical and stereo-chemical properties, and structure-activity and structure-metabolism relationships [ 21 , 22 ]. PCBs are known to sequester into tissues richly supplied by blood flow like intestine wall and this occurs by passive diffusion [ 21 , 22 ]. Since PCBs are known to get absorbed via oral, dermal, and inhalation routes, the main routes of excretion include, obviously, fecal, cutaneous, and respiratory air. PCB congeners are known to accumulate in adipocytes, irrespective of dose and species, apart from blood - hence, their distribution in vivo is determined by the blood flow and lipophilicity directly [ 23 , 24 ].Generally, ortho-substituted PCBs have higher partition coefficients as compared to non-ortho and dioxin-like PCBs which possess all forms of chlorine substitutions [ 25 , 26 ]. PCB 77 being a dioxin-like highly lipid soluble PCB sequesters to brain but cannot be removed easily due to tissue sequestration in brain tissues and cause thyroid and neuroendocrine disruption [ 27 , 28 ]. However, PCBs are known to disrupt Blood Brain Barrier and enter the brain easily on high doses [ 29 , 30 ]. Hepatic binding to cytochrome enzymes also determines their higher hepatic concentrations. PCBs are known to induce the metabolism of other xenobiotics including PCBs themselves. Further fate and accumulation of PCBs are determined by what fraction of the administered congener is metabolized and how fast this process happens. This depends on chemistry of each congener [ 31 ]. Coplanar PCBs which have favorable spatial arrangement for hindering metabolism, and mono-ortho substituted PCBs, do not get metabolized easily [ 30 , 32 ]. Unsubstituted chlorine at para-position changes the ability of the congener to form toxic arene metabolites easily causing free radical damage. [ 21 , 33 ]. Non-substituted PCBs at ortho-meta positions and possession of higher number of chlorine atoms like PCB 180, makes the congener to cumulate, especially after higher or repeated dosing [ 25 , 34 , 35 ]. Elimination kinetics of PCB congeners vary between tissue compartments and the dose-exposure protocol. For example, the half-life of 3,3',4,4'-TCB was1.07–2.60 days, since this congener sequesters in adipocytes, liver and thymocytes [ 19 ]. Congeners which do not tissue-cumulate, like 2,2',5,5'-TCB, havehalf-lives from 1.64–2.90 days. Since their concentrations were similar despite this difference in tissue sequestration, the property that decides eliminationkinetics is therefore potency of congener, apart from chirality of stereochemistry [ 32 ]. PCB 77 being highly more potent than PCB 180 in its toxic effects,makes it selectively cumulate in brain and produce behavioral depression and shorter plasma elimination half-lives [ 36 , 37 ]. The key results for behavioral effects of the PCB mixture include a decline in mood, autonomic and miscellaneous domain sub-scores, in a dose dependentand time dependent manner at each dose level, being contributed significantly by alteration in certain sub domains. The decline in grooming in this study at 2–4 hours after exposure initiation hence is not due to introduction of mice to novel environment of chamber and can be ascribed to be influenced only bythe PCB induced neurotransmitter changes in brain. The negative chronotropic effects at 2 hours of exposure to PCBs may be due to cardiac, reflex, and central depression. This is also the time when PCBs are about to redistribute from blood to the brain after being absorbed from the alveoli. Respiratory rate reduced at 4 hours of daily exposures especially in high dose exposures. Dose-responses with monoamines followed linear (5-HT), non-monotonic (DA) and exponential (NA) relationships till 6 hours of exposure. Conclusion Generally, DA reduced at Tmax for each dose level; 5-HT increased exponentially as Tmax approached for each dose level; and NA followed U-shaped dose response, the dip being correlated at Cmax. PCBs are known to affect monoamine levels in neurons inside and outside the Central Nervous System, via multiple mechanisms, including inhibition of VMAT-2 and reduction of dopamine levels in synapse [ 38 ]. In their ability to alter dopamine levels in brain and cause depression in humans, non-coplanar PCBs are more potent, as in this study wherein the decrease in dopamine and mood were maximum at PCB 77 Tmax. The probable mechanisms of these behavioral effects of PCB 77 inhalation on mood are outlined in Fig. 7, which may be relevant in causing neuropsychiatric illnesses (e.g. major depression, autism, OCD) in human patients. In conclusion, this study proves that PCBs can alter behavior and monoamine levels via inhalation route. Abbreviations 5-HT = 5-hydroxytrytamine; Acetonitrile (ACN); ANOVA= Analysis of Variance; AUC0-t = Area-Under-Plasma-Concentration-time-Curve from time = 0 to time = t; AUC0-∞ = Area-Under-Plasma- Concentration-time-Curve from time = 0 to time = infinity; CB = Chloro-biphenyl; CPCSEA = Committee for the Purpose ofControl and Supervision of Experimentation in Animals; Cmax = Maximum Concentration; CL = Clearance; DA = Dopamine; DAD = Diode Array Detector;EDTA = Ethylene Diamine Tetra Acetate; FLD = Fluorescence detector; HPLC = High-Performance Liquid Chromatography; IAEC = Institutional Animal EthicsCommittee; ID-50 = Inhibitory Dose-50%; IS = Internal Standard; K el = Elimination rate constant; K ow = Log (Octanol-water) partition coefficient; OECD =Organization for Economic Cooperation & Development; MRT = Mean Residence Time; NA = Noradrenaline; PCBs = Polychlorinated Biphenyls; PBS =Phosphate Buffered Saline; QC = Quality Control; RM-ANOVA = Repeated Measures-Analysis of Variance; t1/2 = Half-life; TBME = tertiary-Butyl Methyl Ether;Tmax = Maximum Time; UPLC = Ultra-high Performance Liquid Chromatography; UV = Ultraviolet; Vd = Volume of Distribution. Declarations Ethics approval - The study was carried out after PSG-IAEC approval from a CPCSEA-approved Animal Facility. The study was conducted in accordance with CPCSEA guidelines for laboratory animal facility, 2005. Consent to participate – Not applicable Consent for publication - It has been approved by all the authors listed as per ICJME criteria. Availability of data and materials - The manuscript includes data which is linked to the study results. This data collection can be made available on reasonable and justified request and shall be bound by the publishers’ data management policy for submitting manuscripts involving original research. This data policy does not breach any confidentiality and is based sound ethical compliance. Infact, the supplementary files have been attached for initial review and plausible publication, including review thereafter, to augment and enhance the clarity, reproducibility, and authenticity of the results discussed, with prior approval of all the authors. Preprint available at DOI: https://doi.org/10.21203/rs.3.rs-1425455/v1 Competing interests - The authors declare no conflicts of interests during the conception and inception of the paper, including any non-financial and monetary ones. All the authors have read and approved the manuscript, and all data were generated in-house and that no paper mill was used. Funding - The authors declare that the study was self-funded with no other financial funding contributions / drawing from outside by the authors. Authors' contributions - SSK contributed to manuscript editing in addition to data analysis and visualization and chromatography (new methods). NR contributed to study inception, design, data analysis, manuscript writing and editing and data analysis. Acknowledgements - The authors would like to acknowledge the facilities given by PSG Institute of Medical Sciences & Research, Coimbatore for the success of the work. The authors acknowledge the contributions of the former Dean and Professor of Pharmacology and PSG-CMMT, Dr. Ramalingam Sankaran (formerly), for his administrative and logistic facilities provision, as the Guide of the Part time PhD program of one of the authors. References Stockholm Convention - Home page. http://www.pops.int/default.aspx. Accessed 25 Jan 2022 Health UD of, Services H (1999) Agency for Toxic Substances and Disease Registry-ATSDR. Eriksson P, Fredriksson A (1996) Developmental neurotoxicity of four ortho-substituted polychlorinated biphenyls in the neonatal mouse. Environ ToxicolPharmacol 1:155–165. https://doi.org/10.1016/1382-6689(96)00015-4 Seegal RF (1996) Epidemiological and laboratory evidence of PCB-induced neurotoxicity. Crit Rev Toxicol 26:709–737. https://doi.org/10.3109/10408449609037481 Simon T, Britt JK, James RC (2007) Development of a neurotoxic equivalence scheme of relative potency for assessing the risk of PCB mixtures. RegulToxicolPharmacol 48:148–170. https://doi.org/10.1016/j.yrtph.2007.03.005 Louis C, Covaci A, Crocker DE, Debier C (2016) Lipophilicity of PCBs and fatty acids determines their mobilisation from blubber of weaned northern elephant seal pups. Sci Total Environ 541:599–602. https://doi.org/10.1016/j.scitotenv.2015.09.094 Irwin S (1968) Comprehensive observational assessment: Ia. A systematic, quantitative procedure for assessing the behavioral and physiologic state of the mouse. Psychopharmacologia 13:222–257. https://doi.org/10.1007/BF00401402 Committee for the Purpose of, Animals Supervision of Experiments on Animals (2003) CPCSEA Guidelines for laboratory animal facility. Indian J Pharmacol 35: OECD (2009) OECD guidance document on acute inhalation toxicity testing. The Environment, Health and Safety Division, Organization for Economic Cooperation & Development Narayanan R, Muthusamy S, Lakshmi SV, et al (2021) Design and development of whole-body rodent inhalation chamber for exposure to Aroclor 1232 in Swiss albino mice. Int J Environ Sci Technol. https://doi.org/10.1007/s13762-021-03515-8 Ramanujam N, Sivaselvakumar M, Ramalingam S (2017) Fast and parallel determination of PCB 77 and PCB 180 in plasma using ultra performance liquid chromatography with diode array detection: A pharmacokinetic study in Swiss albino mouse. Biomed Chromatogr31:. https://doi.org/10.1002/bmc.4000 Wang H, Walaszczyk EJ, Li K, et al (2012) High-performance liquid chromatography with fluorescence detection and ultra-performance liquid chromatography with electrospray tandem mass spectrometry method for the determination of indoleamine neurotransmitters and their metabolites in sea lamprey plasma. Anal Chim Acta 721:147–153. https://doi.org/10.1016/j.aca.2012.01.025 Grandjean P, Budtz-Jørgensen E, Barr DB, et al (2008) Elimination half-lives of polychlorinated biphenyl congeners in children. Environ Sci Technol 42:6991–6996. https://doi.org/10.1021/es800778q Carpenter DO (2015) Exposure to and health effects of volatile PCBs. Rev Environ Health 30:81–92. https://doi.org/10.1515/reveh-2014-0074 Seegal RF, Okoniewski RJ, Brosch KO, Bemis JC (2002) Polychlorinated biphenyls alter extraneuronal but not tissue dopamine concentrations in adult rat striatum: an in vivo microdialysis study. Environ Health Perspect 110:1113–1117. https://doi.org/10.1289/ehp.021101113 Matthews HB, Dedrick RL (1984) Pharmacokinetics of PCBs. Annu Rev PharmacolToxicol 24:85–103. https://doi.org/10.1146/annurev.pa.24.040184.000505 Berninger JP, Tillitt DE (2019) Polychlorinated biphenyl tissue-concentration thresholds for survival, growth, and reproduction in fish. Environ Toxicol Chem 38:712–736. https://doi.org/10.1002/etc.4335 Grimm FA, Hu D, Kania-Korwel I, et al (2015) Metabolism and metabolites of polychlorinated biphenyls. Crit Rev Toxicol 45:245–272. https://doi.org/10.3109/10408444.2014.999365 Clevenger MA, Roberts SM, Lattin DL, et al (1989) The pharmacokinetics of 2,2’,5,5’-tetrachlorobiphenyl and 3,3’,4,4’-tetrachlorobiphenyl and its relationship to toxicity. Toxicol Appl Pharmacol 100:315–327. https://doi.org/10.1016/0041-008x(89)90317-7 Lotti M (2003) Pharmacokinetics and blood levels of polychlorinated biphenyls. Toxicol Rev 22:203–215. https://doi.org/10.2165/00139709-200322040-00003 Corrigan FM, Murray L, Wyatt CL, Shore RF (1998) Diorthosubstituted polychlorinated biphenyls in caudate nucleus in Parkinson’s disease. Exp Neurol 150:339–342. https://doi.org/10.1006/exnr.1998.6776 Parham FM, Portier CJ (1998) Using structural information to create physiologically based pharmacokinetic models for all polychlorinated biphenyls. II. Rates of metabolism. Toxicol Appl Pharmacol 151:110–116. https://doi.org/10.1006/taap.1998.8441 Parham FM, Kohn MC, Matthews HB, et al (1997) Using structural information to create physiologically based pharmacokinetic models for all polychlorinated biphenyls. Toxicol Appl Pharmacol 144:340–347. https://doi.org/10.1006/taap.1997.8139 Kodavanti PR, Ward TR, Derr-Yellin EC, et al (1998) Congener-specific distribution of polychlorinated biphenyls in brain regions, blood, liver, and fat of adult rats following repeated exposure to Aroclor 1254. Toxicol Appl Pharmacol 153:199–210. https://doi.org/10.1006/taap.1998.8534 Oberg M, Sjödin A, Casabona H, et al (2002) Tissue distribution and half-lives of individual polychlorinated biphenyls and serum levels of 4-hydroxy-2,3,3’,4’,5-pentachlorobiphenyl in the rat. Toxicol Sci 70:171–182. https://doi.org/10.1093/toxsci/70.2.171 Beck V, Roelens SA, Darras VM (2006) Exposure to PCB 77 induces tissue-dependent changes in iodothyronine deiodinase activity patterns in the embryonic chicken. Gen Comp Endocrinol 148:327–335. https://doi.org/10.1016/j.ygcen.2006.04.003 Simmons SL, Cummings JA, Clemens LG, Nunez AA (2005) Exposure to PCB 77 affects the maternal behavior of rats. PhysiolBehav 84:81–86. https://doi.org/10.1016/j.physbeh.2004.10.022 Jandacek RJ, Rider T, Keller ER, Tso P (2010) The effect of olestra on the absorption, excretion and storage of 2,2’,5,5’ tetrachlorobiphenyl; 3,3’,4,4’tetrachlorobiphenyl; and perfluorooctanoic acid. Environ Int 36:880–883. https://doi.org/10.1016/j.envint.2009.06.010 Seelbach M, Chen L, Powell A, et al (2010) Polychlorinated biphenyls disrupt blood-brain barrier integrity and promote brain metastasis formation. Environ Health Perspect 118:479–484. https://doi.org/10.1289/ehp.0901334 Quinete N, Esser A, Kraus T, Schettgen T (2017) PCB 28 metabolites elimination kinetics in human plasma on a real case scenario: Study of hydroxylated polychlorinated biphenyl (OH-PCB) metabolites of PCB 28 in a highly exposed German Cohort. Toxicol Lett 276:100–107. https://doi.org/10.1016/j.toxlet.2017.05.025 Kania-Korwel I, Lehmler H-J (2016) Chiral polychlorinated biphenyls: absorption, metabolism and excretion--a review. Environ Sci Pollut Res Int 23:2042–2057. https://doi.org/10.1007/s11356-015-4150-2 Dhakal K, Gadupudi GS, Lehmler H-J, et al (2018) Sources and toxicities of phenolic polychlorinated biphenyls (OH-PCBs). Environ Sci Pollut Res Int 25:16277–16290. https://doi.org/10.1007/s11356-017-9694-x Haraguchi K, Koga N, Kato Y (2005) Comparative metabolism of polychlorinated biphenyls and tissue distribution of persistent metabolites in rats, hamsters, and Guinea pigs. Drug MetabDispos 33:373–380. https://doi.org/10.1124/dmd.104.002444 Danis B, Cotret O, Teyssié JL, et al (2004) Coplanar PCB 77 uptake kinetics in the sea star Asterias rubens and subsequent effects on reactive oxygen species (ROS) production and levels of cytochrome P450 immunopositive proteins (CYP1A-IPP). Marine Ecology Progress Series 279:117–128. https://doi.org/10.3354/meps279117 Kania-Korwel I, Lehmler H-J (2016) Toxicokinetics of chiral polychlorinated biphenyls across different species--a review. Environ Sci Pollut Res Int 23:2058–2080. https://doi.org/10.1007/s11356-015-4383-0 Tan Y, Song R, Lawrence D, Carpenter DO (2004) Ortho-substituted but not coplanar PCBs rapidly kill cerebellar granule cells. Toxicol Sci 79:147–156. https://doi.org/10.1093/toxsci/kfh108 Viluksela M, Heikkinen P, van der Ven LTM, et al (2014) Toxicological profile of ultrapure 2,2’,3,4,4’,5,5’-heptachlorbiphenyl (PCB 180) in adult rats. PLoS ONE 9:e104639. https://doi.org/10.1371/journal.pone.0104639 Bemis JC, Seegal RF (2004) PCB-induced inhibition of the vesicular monoamine transporter predicts reductions in synaptosomal dopamine content. Toxicol Sci 80:288–295. https://doi.org/10.1093/toxsci/kfh153 Tables Table 1: The Mean Pharmacokinetic Parameters of PCB 77 in Plasma and Brain after inhalation of PCBs at 2, 4 and 16 mg/m 3 dose Pharmacokinetic Parameters PCB 77 Plasma Brain 2.0 mg/cu.m (mean±SD, N=4) 4.0 mg/cu.m (mean±SD, N=4) 16.0 mg/cu.m (mean±SD, N=4) 2.0 mg/cu.m (mean±SD, N=4) 4.0 mg/cu.m (mean±SD, N=4) 16.0 mg/cu.m (mean±SD, N=4) C max (ng/mL) 88.32±8.28 245.62±39.58 727.40±42.52 69.43±10.54 175.13±6.42 526.21±28.63 T max (hr) 3.50±1.00 4.00±0.00 4.00±0.00 3.50±1.00 4.00±0.00 2.00±0.00 AUC 0-t (ng.hr/mL) 1321.19±55.72 2749.52±392.00 7565.21±148.44 1254.97±57.51 2366.40±142.19 6798.93±340.47 AUC 0-∞ ( ng.hr/mL) 1626.02±64.13 2957.28±413.20 8013.39±263.11 1469.62±161.80 2581.12±136.46 7100.56±413.72 T 1/2 (hr) 20.17±1.38 12.72±0.36 11.17±1.54 16.20±4.36 14.20±0.58 10.82±1.29 MRT last 16.51±0.42 14.31±0.49 14.29±0.24 17.07±1.93 15.05±0.32 14.39±0.30 K el (hr -1 ) 0.03± 0.00 0.06±0.00 0.06±0.01 0.05±0.01 0.05±0.00 0.07±0.00 Vd (ml.m -2 ) 35858.25±3073.55 25209.01±3835.06 32099.78±3593.04 31481.64±5927.66 31856.13±3012.73 35070.00±2110.32 CL (ml.hr -1 m -2 ) 1231.46±49.43 1375.07±215.15 1998.28±65.96 1373.32±150.80 1553.12±85.97 2258.90±127.06 Table 2: Tabulation of mean monoamine concentrations at T max after exposure to different doses of Aroclor 1232 5-HT (ng/ml) 2 mg/cu.m 4 mg/cu.m 16 mg/cu.m Mean SD Mean SD Mean SD 2 hour 2.07 0.07 2.11 0.13 3.08 0.23 4 hour 3.04 0.27 2.62 0.22 4.32 0.47 6 hour 4.33 0.67 3.29 0.23 4.36 0.18 DA (ng/ml) 2 mg/cu.m 4 mg/cu.m 16 mg/cu.m 2 hour 2.99 0.54 1.75 0.45 3.39 1.04 4 hour 2.42 0.64 0.83 0.13 3.73 0.48 6 hour 5.15 0.85 4.91 0.51 5.07 0.46 NA (ng/ml) 2 mg/cu.m 4 mg/cu.m 16 mg/cu.m 2 hour 0.24 0.08 0.27 0.16 0.23 0.15 4 hour 1.88 0.25 0.85 0.11 1.01 0.09 6 hour 5.28 0.67 5.13 1.35 2.15 0.15 Table 3: Behavioral effects of 4 mg/cu.m dose level of Aroclor 1232 at PCB 77 T max in brain. BEHAVIOURAL DOMAIN BEHAVIOURAL SUB-DOMAIN EFFECTS OF PCB 77 AT 4 Hrs. (4 mg/cu.m of AROCLOR 1232) AWARENESS ALERTNESS VISUAL PLACING PASSIVITY STEREOTYPY MOOD GROOMING VOCALIZATION RESTLESSNESS IRRITABILITY / AGGRESSION FEARFULNESS MOTOR ACTIVITY REACTIVITY TO ENVIRONMENT SPONTANEOUS ACTIVITY TOUCH RESPONSE PAIN RESPONSE CNS EXCITATION STARTLE RESPONSE STRAUB TAIL TREMORS TWITCHES CONVULSIONS POSTURE BODY POSTURE LIMB POSITION MOTOR INCOORDINATION STAGGERING GAIT ABDOMINAL GAIT RIGHTING REFLEX MUSCLE TONE LIMB TONE GRIP STRENGTH BODY SAG BODY TONE ABDOMINAL TONE REFLEXES PINNA CORNEAL IFR (Ipsilateral flexor Reflex) AUTONOMIC WRITHING PUPIL SIZE PALPEBRAL OPENING EXOPHTHALMOS URINATION SALIVATION PILOERECTION HYPOTHERMIA SKIN COLOUR HEART RATE RESPIRATORY RATE MISCELLANEOUS EXPLORATORY ACTIVITY HOSTILITY SOCIAL BEHAVIOR LETHALITY / DEAD ACUTE DELAYED Colour-coding: No change absolutely in all animals CNS Modulatory change in all animals CNS depression in 50% animals CNS excitation in 50% animals Mild change in 50% of animals CNS modulation in 50% animals CNS depression in all animals CNS excitation in all animals Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4715900","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":327660047,"identity":"f606d80a-67d4-4292-baca-b4a59e8ea392","order_by":0,"name":"Sivaselvakumar Muthusamy","email":"","orcid":"","institution":"PSG Institute of Medical Sciences \u0026 Research","correspondingAuthor":false,"prefix":"","firstName":"Sivaselvakumar","middleName":"","lastName":"Muthusamy","suffix":""},{"id":327660048,"identity":"1b0e0025-c319-4529-a34d-10a28fed907c","order_by":1,"name":"Ramanujam Narayanan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4klEQVRIiWNgGAWjYLCChAoJOX7+5gNApoQMcVoenLEwlpxxLAGkhYcoHYwP2yoSNxzIMQBxCGsxbz/8dENim0TizIYzn1/dqLHgYWA/fHQDPi0yZ9LMbiSckzDuZ+7dZp1zDOgwnrS0G/i0SDAkALWUScjObDi7zTiHDahFgscMvxb+599uJLBJMAL98sw45x8xWiRygLa0SSgCtTA/zm0jSsubshsJZyRAgWzGnNsnwcNG0C/86dtu/qioA0Xl488534AM9sPH8GpBBmwSYJJY5SDA/IEU1aNgFIyCUTByAAARM0ynqB+bcwAAAABJRU5ErkJggg==","orcid":"","institution":"PSG Institute of Medical Sciences \u0026 Research","correspondingAuthor":true,"prefix":"","firstName":"Ramanujam","middleName":"","lastName":"Narayanan","suffix":""}],"badges":[],"createdAt":"2024-07-10 06:08:18","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4715900/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4715900/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":61793291,"identity":"f00276e5-9d06-438c-ae3c-f59e7f18c384","added_by":"auto","created_at":"2024-08-05 16:09:26","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":230699,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"03.Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-4715900/v1/0fa1bce3a1bde2242a0a1311.png"},{"id":61793284,"identity":"1f875d0c-e38a-4501-a9ac-f2cf31cac25c","added_by":"auto","created_at":"2024-08-05 16:09:18","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":108035,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"04.Fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-4715900/v1/6be2042d4b3901b1bb04b4d3.png"},{"id":61793296,"identity":"05622f03-b3b1-42a9-8f59-f942102f9556","added_by":"auto","created_at":"2024-08-05 16:09:34","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":176234,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"05.Fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-4715900/v1/7ef22deb7f03e722bfe8c7b4.png"},{"id":61793289,"identity":"9bb8ad91-849a-4a32-99dc-b30580255c46","added_by":"auto","created_at":"2024-08-05 16:09:24","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":289291,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"06.Fig4.png","url":"https://assets-eu.researchsquare.com/files/rs-4715900/v1/f6d2b4e685ecd838be99b117.png"},{"id":61793294,"identity":"1775d2f3-1b00-4673-b583-dca828128d2e","added_by":"auto","created_at":"2024-08-05 16:09:29","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":894482,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"07.Fig5.png","url":"https://assets-eu.researchsquare.com/files/rs-4715900/v1/05b5a4e3b99d22a490ded519.png"},{"id":61793283,"identity":"0f5426de-c81d-46e4-9226-9edfa0068e90","added_by":"auto","created_at":"2024-08-05 16:09:17","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":92440,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"08.Fig6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4715900/v1/08af62a8e69ea6cf5ff7c92f.jpg"},{"id":85359601,"identity":"4aa2f9cc-cfe4-4782-88e8-731b3ca17b58","added_by":"auto","created_at":"2025-06-25 05:46:48","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3850817,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4715900/v1/f9bfbfab-b23b-4509-835b-09e1a56cdb51.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Behavioral Toxicodynamics and Kinetic profiling of PCB congeners in Swiss albino mouse dosed with Aroclor 1232 through inhalational route: A pivotal risk assessment study","fulltext":[{"header":"Introduction","content":"\u003cp\u003ePoly-Chlorinated Biphenyls (PCBs) are a chemical class of 209 congeners that are known to bio-accumulate, bio-magnify, bio-concentrate and persistacross all the levels of the food chain. Though the production, distribution, and supply of their commercial mixtures like Aroclors and Kaneclors have beenstrictly regulated in most of the member countries by the United Nations Stockholm Convention [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e] their replacement by best alternatives and their properdisposal are not practically feasible in many regions regulated. This is due to their favourable thermal inertness, hydraulic and dielectric nature, because ofwhich their use in electronics, rubber and plastic industries are unavoidable; additionally, higher lipophilicity and halogenated nature make theirenvironmental elimination practically impossible [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe 209 chemical congeners of PCBs can be further classified based on the number of chlorine substitutions, as low chlorine (1\u0026ndash;4 chlorinated biphenyls)and high chlorine (5\u0026ndash;10 chlorinated biphenyls) homolog families, each family having one to many congeners differing in the specific spatial chlorinesubstitution on the biphenyl nucleus. The number and the exact site of the chlorine substitution determine the nature of the congener\u0026rsquo;s structure-activityrelationship and its metabolism in-vivo, which determine its pharmacokinetics, pharmacodynamics, cumulation, and toxicological profiles. For example,ortho-substituted PCBs are known to be highly neurotoxic [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e], whereas co-planar PCBs are known to possess cardiometabolic toxicities [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Similarly, DioxinlikePCBs are proved to have potential to bind to Aryl-Hydrocarbon receptors in the cell nucleus and cause neuroendocrine disruption [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Highly chlorinatedPCB congeners are known to be highly lipophilic and persist in the environment, with long biological half-lives [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. However, the complex individualchemistry of these congeners and the variable molar composition of their restricted commercial mixtures make the predictability of the biologicalpharmacokinetics of PCB congeners after dosing of these mixtures complicated and difficult in-vivo. Aroclor 1232 is a semi-volatile commercial PCBmixture that was selected as the test substance for its ability to cause occupational air pollution indoor and outdoor. It is low chlorinated (32% Chlorine bymolar mass) \u0026ndash;hence, may not have long environmental half-lives despite in-vivo accumulation. It also has low vapor pressure such that it can also leachsoil from air and be carried out to water bodies [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. PCB 77 is a Dioxin like tetra-chloro-biphenyl (Tetra CB) that is also semi-volatile but is highly potent incausing neuroendocrine toxicities after binding to Aryl Hydrocarbon receptors [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. PCB 180 is a highly lipophilic hepta-CB congener that is known to persistin biological systems and cause cardio-metabolic toxicities [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. PCB 180 has higher octanol-water partition coefficient than PCB 77 [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAs a blind screening tool, the Irwin neurobehavioral scale is a validated scale to be used to assess the cross sectional behavioral, autonomic, neurological,and physiological status of the laboratory mouse, both in normal and treated animals [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Additionally, it can be used to delineate the pharmacologicaleffects of diverse groups of drugs acting on the nervous system. Using existing HPLC-FLD-based approaches for measurement of monoamines, the studyevaluated the behavioral changes with pharmacokinetic time-points in relation to monoamine concentration changes.\u003c/p\u003e \u003cp\u003eThis study aims to predict the pharmacokinetic variables of PCB 77 and PCB 180 after inhalation dosing of a PCB commercial mixture, viz., Aroclor 1232, in mice housed in a whole-body inhalation chamber, in an acute exposure protocol. The study then enables to study further the relationships of the PCB 77and PCB 180 levels in brain and plasma with behavior scores assessed by Irwin scale. After dosing this mixture in mice via inhalation chamber, thebehavioral effects of PCB 77 and PCB 180 have been determined using estimations of their plasma and brain concentration-ratios at geometricallyprogressive dose increments.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cstrong\u003eChemicals\u003c/strong\u003e: Aroclor 1232 for inhalation exposure was purchased from Sigma-Aldrich Inc., India. Reference standards PCB 77, PCB 180, PCB 101, 5-Hydroxytryptamine (5-HT), Dopamine (DA), Nor-adrenaline (NA) and Paracetamol were obtained from Sigma-Aldrich Inc., India. PCB 101 was used as an Internal Standard (IS). Solvents including methanol, tert-Butyl Methyl Ether (TBME) and acetonitrile (ACN) were purchased from Sigma-Aldrich Inc., India. High-purity water for chromatographic analyses was purchased from Merck-Millipore Inc., India.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAnimals:\u0026nbsp;\u003c/strong\u003eInstitutional Animal Ethics Committee (IAEC) approval was obtained prior to the conduct of the study for utilization of Swiss albino mice, which has been recognized by the Indian Regulatory authority,the Committee for the Purpose of Control and Supervision of Experiments on Animals [8]. All mice were\u003c/p\u003e\n\u003cp\u003eFemales about 16-32 gm at initiation and were housed in pairs per cage. Before study initiation, all the animals were acclimatized, and allocated to the three dose levels randomly. Except during the exposure period, they were provided with water and feed ad-libitum and housed under standard circadian rhythm and housing conditions as per OECD guidance for using humane endpoints in toxicity studies as applied to inhalation toxicity [9].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eInhalation Chamber and the Exposure protocol\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eComponents of a whole-body inhalation chamber which was used for exposure of test chemical in mice consisted of five units \u0026ndash; the inlet, the animal exposure-control unit, the outlet, the connecting lines, and the external indoor environment [10]. Exposures to Aroclor 1232 consisted of three test doses in geometric progression (2.0, 4.0 and 16.0 mg.m-3). Each dose of the test substance was given to a group of 36 mice in the inhalation chamber as per OECD guidelines for sub-acute inhalation toxicity [9]. A cohort of four animals per time point was procured out from the chamber for each time point (pre-dose, 2.0, 4.0, 6.0, 8.0, 12.0, 24.0, 36.0, 48.0 hours) for a total of nine time-points per dose level. The internal chamber environmental variables like oxygen content, relative humidity, temperature, pressure, and flow rate of air through the inlet along with the Aroclor 1232 solution were monitored and controlled for ensuring proper animal welfare.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eIrwin Behavior assessment\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIt required about three to eight minutes of testing per mouse for its behavioral rating scores using Irwin scale. The scale consists of about forty-seven different functions and processes, divided into eleven domains, each affecting the central, peripheral, and somatic, and the autonomic nervous system, with effects related to cardiovascular and respiratory systems. Certain of these functions or sub domains can be tested by observation, whereas others can be assessed in the mouse after gentle maneuvers. Each sub domain has a minimum score, a normative score, and a maximum score, ranging from 0-4.\u003c/p\u003e\n\u003cp\u003eThe initial undisturbed observation consisted of assessment of body position, locomotor activity, exophthalmos, respiration, tremors, twitches, convulsions, exploratory activity, hostility, and social conglomeration, as the mice remained inside the chamber. The animal was scored on a scale of 0-4 for each of these sub domain functions. After completion of this initial observation, the mouse was transferred from the chamber into the cage and then gently manipulated towards the floor of the chamber for response grading to assess the transfer arousal, spatial locomotion, palpebral closure, startle response, piloerection, tremor grading, gait and limb rotation. Following these scoring steps, the animal is lifted vertically by its mid tail above a wired mesh grid, and lowered slowly for assessment of visual placing response, i.e., extension of all limbs before contact. Grip strength and positional struggle behavior were then assessed. Body tone, gross signs of hypothermia, pinna and corneal reflexes, toe-pinch response were then graded. After this, the mouse was transferred to the chamber and graded for alley progression, withdrawal responses, touch-escape response, and positional passivity. Finally, autonomic signs like diarrhea, skin / fur colour, limb tone, abdominal tone, pupillary reflex, lacrimation, salivation, pupillary size, and provoked biting response (for\u003c/p\u003e\n\u003cp\u003eaggression) were observed followed by tail-pinch response, righting reflex, provoked freezing, grasp irritability, vocalization and defecation and urination assessments [7].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eSample collection\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods of Euthanasia:\u0026nbsp;\u003c/strong\u003eEach animal was euthanized with Ketamine (55-77 mg/kg intraperitoneally) at the sacrifice as per AVMA Guidelines.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods of Anesthesia / Analgesia and Humane endpoints for animal welfare:\u0026nbsp;\u003c/strong\u003eAll mice were observed for vital parameters like Heart rate, Respiratory rate and involuntary movements after ketamine overdose administration. If tachycardia (HR \u0026gt;2 beats / s) / tachypnoea (RR \u0026gt; 5 /10 sec) / involuntary movements (Flexion / extension of limbs) were present even after Ketamine overdose, the animal was given Xylazine 4 mg/kg overdose I.P. immediately to avoid suffering. As per Refinement (Russel \u0026amp; Burch\u0026rsquo;s 3Rs) this step alleviates animal suffering and conforms to humane endpoints of animal welfare. Anesthesia \u0026amp; analgesia were not used for the study as they are not compatible with the study protocol. Death was immediate with Xylazine overdose (within next 2 minutes).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSpecimen / sample coding\u003c/strong\u003e: All animal samples were collected in coded sample tubes as plasma or brain in 1x PBS (ice cold). The codes were later decoded only upon statistical analysis.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePlasma collection and processing:\u0026nbsp;\u003c/strong\u003eBefore euthanizing PCB-exposed mice at each time point for dissection and homogenization of brain samples by high dose Ketamine (55 mg/kg I.P.) (Leary et al., 2013), their blood samples were collected by a terminal cardiac puncture (maximum 0.2 ml) in tubes premixed with 0.012 M buffered calcium disodium EDTA in the ratio of 3:1 of anticoagulant solution: blood, and these were then centrifuged at 3000 rpm for 8 minutes at 4\u0026deg;C immediately. The centrifuged sample volumes were then separated for their plasma component, and the plasma was stored at -80 \u0026plusmn; 10\u0026deg;C, until analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBrain collection and processing:\u0026nbsp;\u003c/strong\u003eThe euthanized mice brains were dissected out as whole; homogenized for storage in 1% of ice-cold Phosphate Buffered Saline (PBS) solution in -80 \u0026plusmn; 10\u0026ordm;C until analysis. Blank brain samples were collected at 0 hours pre-dosing for calibration purposes. Each brain sample was divided into two equal portions for estimation of PCB levels and monoamine levels, respectively.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInstrumentation and Chromatography conditions for PCB estimation:\u0026nbsp;\u003c/strong\u003eThe Ultra-High-Performance Liquid Chromatography (UPLC) instrument consisted of a Waters Acquity H class UPLC system equipped with a quaternarypump and 96-vial autosampler coupled with a diode array UV detector (Waters, Milford, MA, USA). The chromatographic separation was performed on anAcquity UPLC BEH C18 column from Waters (2.1 mm \u0026times; 100mm; 1.7\u0026mu;m). The column temperature was set at 35\u0026ordm;C and the autosampler was kept at 15\u0026ordm;C.\u003c/p\u003e\n\u003cp\u003eThe mobile phase composed of a mixture of acetonitrile (85%, v/v) and water (15%, v/v) at a flow rate of 0.3ml/min. Before analysis, the mobile phase wasfiltered through a 0.22\u0026mu;m membrane filter and degassed by ultra-sonication. A 5\u0026mu;l injection of each sample was loaded on to the system and total analysistime was 6 min. The Diode Array Detector (DAD) was set at 215 nm. The sampling needle was washed with 300\u0026mu;l of strong wash (ACN/water, 65/35) toreduce carry-over and 300\u0026mu;l of weak wash (ACN/water, 30/70). Data acquisition was done using Empower 3 software version 1.0 (Waters, USA) [11].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eParent monoamine analysis measurements:\u0026nbsp;\u003c/strong\u003e5-HT, DA and NA stock solutions of 1mg/mL were prepared by dissolving a suitable amount of single standard in acetonitrile. Mixture of stock solutions 5-HT, DA and NA (100 \u0026mu;g/mL) were prepared in acetonitrile. The High-Performance Liquid Chromatography (HPLC) instrument consisted of a Waters 515HPLC system equipped with an isocratic pump and coupled with a fluorescence detector (Waters, Milford, MA, USA). The chromatographic separation wasperformed on a Gemini-C18 column from Phenomenex (4.3 mm \u0026times; 50mm; 3\u0026mu;m).Buffer was prepared using 70 mM potassium dihydrogen phosphate, 0.1mM dipotassium EDTA salt, 1.1 mM heptane sulfonic acid and 0.4 % Trifluoracetic acid added to 1000 mL water and adjusted to a pH of 3.14 usingsaturated citric acid solution. The mobile phase composed of a mixture of methanol (10%, v/v) and buffer (90%, v/v) at a flow rate of 0.5ml/min.Paracetamol used as internal standard (IS). The mobile phase was filtered through a 0.22\u0026mu;m membrane filter and degassed by ultra-sonication. Thefluorescence detector was set excitation of 285 nm and emission at 345 nm respectively. Data acquisition was done using Empower 2 software version 1.0(Waters, USA) [12].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis:\u0026nbsp;\u003c/strong\u003eData were represented as mean \u0026plusmn; standard deviation. Win-Non-Lin software 5.1 was used to calculate the pharmacokinetic parameters by noncompartmentalmodel (Pharsight Corporation, CA, and USA). Pearson correlation was used to assess the relationships between behavioral scores in eachsub-domain and the time-points for each dose level. One-way Repeated Measures ANOVA was used to evaluate the significance of each correlationcoefficient obtained pair wise in Pearson correlation test. Hill model was used to model dose-response-effects versus brain : plasma ratios for each congenerestimated using Inhibitory-Dose (ID-50) modeling. Pearson Correlation, one-way Repeated Measures Analysis of Variance (RM-ANOVA) and Dose responsemodeling (Hill model, and monoamine analysis) were performed using GraphPad Prism version 8.4.2 for Windows, GraphPad Software, San Diego,California USA, www.graphpad.com.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThe pharmacokinetics of PCB 77 and PCB 180 in plasma and brain were similar with small but notable differences in a dose-dependent manner(Supplemental files 1A and 1B). These parameters could be correlated to central behavioral effects based on brain pharmacokinetics, and autonomicnervous system effects based on plasma pharmacokinetics, in dose-dependent and time-dependent manner (Supplemental files 3A and 3B). Generally,brain accumulation of PCB 77 was more significantly responsible for behavioral disruption than PCB 180 accumulation in plasma – based on brain-plasmaratios of both these congeners (Supplemental files 2A and 2B).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePharmacokinetics profile of PCB 77 at 2.0, 4.0 and 16.0 mg.m\u003csup\u003e-3\u003c/sup\u003e in mice plasma\u003c/strong\u003e:The mean maximum concentration attained, Cmax of PCB 77 in plasma was 88.32 ± 8.28 ng.ml\u003csup\u003e-1\u003c/sup\u003e for the 2 mg.m\u003csup\u003e-3\u003c/sup\u003e group. The time taken to achieve this value of Cmax in the plasma of low dose exposure group was 3.50 ± 1.00 hr. The AUC\u003csub\u003e0-t\u0026nbsp;\u003c/sub\u003efor PCB 77 in plasma was 1321.19 ± 55.72 ng.hr.ml\u003csup\u003e-1.\u003c/sup\u003e The Area Under the plasma Concentration-time (infinity) Curve, AUC\u003csub\u003e0-∞\u003c/sub\u003e in plasma was 1626.02 ± 64.13 ng.hr.ml\u003csup\u003e-1\u003c/sup\u003e for PCB 77 after dosing 2 mg.m\u003csup\u003e-3\u003c/sup\u003e of Aroclor 1232. The elimination rate constant, K\u003csub\u003eel\u003c/sub\u003e in plasma for PCB 77 after dosing mice with 2 mg.m\u003csup\u003e-3\u003c/sup\u003e of Aroclor 1232 by inhalation route was 0.03 hr-1. Plasma half-life of PCB 77 in this exposure group was 20.17 ± 1.38 hr. The Mean plasma Residence Times (MRT) for PCB 77 in this dose level was 16.51 ± 0.42 hr. The plasma apparent volume of distribution (Vd) and clearance (CL) for PCB 77 after dosing Aroclor 1232 in this group of mice were 35858.25 ± 3073.55 and, 1231.46 ± 49.43 ml.hr\u003csup\u003e-1.m-2\u003c/sup\u003e, respectively.\u003c/p\u003e\n\u003cp\u003eIn the 4 mg.m-3 exposure group, the Cmax and Tmax of PCB 77 in plasma were 245.62 ± 39.58 ng.ml\u003csup\u003e-1\u003c/sup\u003e and 4.00 hr, respectively. The AUC\u003csub\u003e0-t\u0026nbsp;\u003c/sub\u003eand AUC\u003csub\u003e0-∞\u003c/sub\u003e of PCB 77 in this intermediate dosing group were 2749.28 ± 392.00 and 2957.28 ± 413.20 ng.hr.ml-1 in plasma. The Kel values were 0.06 hr-1 in plasma of themice treated with 4 mg.m\u003csup\u003e-3\u003c/sup\u003e group, while the half-life was 12.72 ± 0.36 hr in this group. The MRT in the intermediate dose exposure groups was 14.31 ± 0.49hr in the plasma. Finally, the Vd and CL in plasma of this exposure level were 25209.01 ± 3835.06 ml.m\u003csup\u003e-2\u0026nbsp;\u003c/sup\u003eand 1375.07 ± 215.15 ml.hr\u003csup\u003e-1.m-2\u003c/sup\u003e.In the 16 mg.m\u003csup\u003e-3\u003c/sup\u003e group, the Cmax of PCB 77 was 727.40 ± 42.52 ng.ml-1 in plasma and Tmax was 4.00 hr. The mean AUC\u003csub\u003e0-t\u003c/sub\u003e and AUC0-∞ in the plasma of thishigh dose group were 7565.21 ± 148.44 and 8013.39 ± 263.11 ng.hr.ml\u003csup\u003e-1\u003c/sup\u003e. In the high dose group, the K\u003csub\u003eel\u0026nbsp;\u003c/sub\u003eand t-1/2 in plasma were 0.06 hr\u003csup\u003e-\u003c/sup\u003e1, and 11.17 ± 1.54hr, respectively. The MRT of PCB 77 in plasma of the mice treated with high dose of Aroclor 1232was 14.29 ± 0.24 hr. The high dose group Vd and CL in plasma was 32099.78 ± 3593.04 ml.m-2 and 1998.28 ± 65.96 ml.hr\u003csup\u003e-1\u003c/sup\u003e.m\u003csup\u003e-2\u003c/sup\u003e in plasma.The mean plasma concentration-time profile of PCB 77 in mouse plasma after inhalation of PCB mixture at three different dose levels are presented inFigure 1, inset A, and Table 1.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePharmacokinetics profile of PCB 77 at 2.0, 4.0 and 16.0 mg.m-3 in mice brain\u003c/strong\u003e:The mean maximum concentration attained; Cmax of PCB 77 was 69.43 ± 10.54 ng.ml\u003csup\u003e-1\u003c/sup\u003e in brains of mice treated with 2 mg.m\u003csup\u003e-3\u003c/sup\u003e of Aroclor 1232 via inhalation route. The Tmax of PCB 77 in brain tissues of mice exposed to this dose was 3.50 ± 1.00 hr. The AUC0-t for PCB 77 in brain was 1254.97 ± 57.51ng.hr.ml\u003csup\u003e-1\u003c/sup\u003e. The AUC0-∞ was 1469.62 ± 161.80 ng.hr.ml\u003csup\u003e-1\u003c/sup\u003e for PCB 77. The elimination rate constant, Kel in brain for PCB 77 was 0.05 hr-1. Brain half-life ofPCB 77 in this exposure group was 16.20 ± 4.36 hr. The Mean brain Residence Times (MRT) for PCB 77 in this dose level was 17.07 ± 1.93 hr. The valuesfor the apparent volume of distribution and tissue clearance of PCB 77 in brain were 31481.64 ± 5927.66 ml.m\u003csup\u003e-2\u003c/sup\u003e respectively, and 1373.32 ± 150.80 ml.hr\u003csup\u003e-1\u003c/sup\u003e.m\u003csup\u003e-\u003c/sup\u003e2, respectively.In the 4 mg.m\u003csup\u003e-3\u003c/sup\u003e exposure group, the Cmax and Tmax of PCB 77 in brain were 175.13 ± 6.42 ng.ml\u003csup\u003e-1\u003c/sup\u003e, and 4.00 hr, respectively. The AUC\u003csub\u003e0-t\u0026nbsp;\u003c/sub\u003eand AUC\u003csub\u003e0-∞\u003c/sub\u003e of PCB77 in this group were 2581.12 ± 136.46 and 2366.40 ± 142.19 ng.hr.ml-1, respectively. The K\u003csub\u003eel\u003c/sub\u003e values were 0.05 hr-1 in brain of the mice treated with 4 mg.m\u003csup\u003e-3\u003c/sup\u003e group, while the half-lifeof PCB 77 in brain was 14.20 ± 0.58 hr in brain. The MRT in the intermediate dose exposure group was 15.05 ± 0.32 hr in brain, while the Vd and CL in brain of this exposure group were 31856.13 ± 3012. 73 ml.m-2 and 1553.12 ± 85.97 ml.hr\u003csup\u003e-1\u003c/sup\u003e.m\u003csup\u003e-2\u003c/sup\u003e.In the 16 mg.m\u003csup\u003e-3\u003c/sup\u003e group, the Cmax of PCB 77 was 526.21 ± 28.63 ng.ml\u003csup\u003e-1\u003c/sup\u003e in brain and Tmax was 4.00 hr. The mean AUC\u003csub\u003e0-t\u003c/sub\u003e and AUC\u003csub\u003e0-∞\u003c/sub\u003e in the brain of thisgroup were 6798.93 ± 340.47 ng.hr.ml\u003csup\u003e-1\u003c/sup\u003eand 7100.56 ± 413.77ng.hr.ml\u003csup\u003e-1.\u003c/sup\u003e The K\u003csub\u003eel\u003c/sub\u003e and t-1/2 in brain were 0.07 hr-1, and 10.82 ± 1.29 hr, respectively. The MRTwas 14.39 ± 0.30 hr. The Vd and CL in brain were 35070.00 ± 2110.32 ml.m-2 and 2258.90 ± 127.06 ml.hr\u003csup\u003e-1\u003c/sup\u003e.m\u003csup\u003e-2\u003c/sup\u003e.The mean brain concentration-time profile of PCB 77 in mouse brain after inhalation of PCB mixture at three different dose levels are presented in Figure 1,inset B, and Table 1.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePharmacokinetics profile of PCB 180 at 2.0, 4.0 and 16.0 mg.m\u003csup\u003e-3\u003c/sup\u003e in mice plasma\u003c/strong\u003e: The mean Cmax of PCB 180 in plasma after 2 mg.m\u003csup\u003e-3\u003c/sup\u003e inhalation of Aroclor 1232 was 66.49 ± 5.22 ng.ml\u003csup\u003e-1\u003c/sup\u003e. The time taken to reach this value of Cmax in plasma was 2.00 hr. The AUC0-t and AUC0-∞ attained by PCB 180 in plasma in this dose level were 763.27 ± 38.80 and 1226.52 ± 136.72 ng.hr.ml\u003csup\u003e-1\u003c/sup\u003e,respectively. The Elimination rate constant of PCB 180 for its elimination from plasma at this dose level was 0.02 hr\u003csup\u003e-1\u003c/sup\u003e. Its half-life in plasma was 28.95 ±3.39 hr. Its MRT in plasma was 13.11 ± 0.96 hr at this dose level. The plasma Vd and CL of PCB 180 after administration of 2 mg.m\u003csup\u003e-3\u003c/sup\u003e of Aroclor 1232 for 6hr were 68083.41 ± 1151.95 ml.m\u003csup\u003e-2\u003c/sup\u003e, and 1646.03 ± 184.36 ml.hr\u003csup\u003e-\u003c/sup\u003e1.m\u003csup\u003e-2\u003c/sup\u003e.The mean Cmax of PCB 180 in plasma in the 4 mg.m\u003csup\u003e-3\u003c/sup\u003e group was 370.21 ± 42.59 ng.ml\u003csup\u003e-1\u003c/sup\u003e, and the time taken to reach this concentration in plasma (Tmax) was 2.00 hr. The AUC\u003csub\u003e0-t\u003c/sub\u003e and AUC\u003csub\u003e0-∞\u003c/sub\u003e were 4369.75 ± 730.55 and 5425.39 ± 408.95 ng.hr.ml\u003csup\u003e-1\u003c/sup\u003e. This congener’s elimination rate constant and half-life were0.04 ± 0.01 hr\u003csup\u003e-1\u003c/sup\u003e and 20.04 ± 6.43 hr, respectively. MRT in plasma for this group was 15.43 ± 0.85 hr; with a Vd and plasma CL of 21729.82 ± 8734.97 ml.m\u003csup\u003e-2\u003c/sup\u003eand 740.51 ± 57.37 ml.hr\u003csup\u003e-1\u003c/sup\u003e.m-2, both somewhat less compared to the lower dose level.In the 16 mg.m\u003csup\u003e-3\u003c/sup\u003e exposure, the time taken to reach a plasma Cmax of 829.57 ± 31.40 ng.ml\u003csup\u003e-1\u003c/sup\u003e was 6.00 hr. The corresponding AUC\u003csub\u003e0-t\u003c/sub\u003e and AUC\u003csub\u003e0-∞\u003c/sub\u003e for theplasma-concentration-time relationship in this group were 13192.39 ± 498.68 and 15636.03 ± 1053.10 ng.hr.ml\u003csup\u003e-1\u003c/sup\u003e. The K\u003csub\u003eel\u0026nbsp;\u003c/sub\u003eand t-1/2 in this group were 0.04 ±0.01 hr-1 and 18.21 ± 1.94 hr, respectively, and the MRT was 16.24 ± 0.32 hr. The Vd and CL of PCB 180 from plasma at this dosing level were 26825.47 ± 1287.33 ml.m\u003csup\u003e-2\u003c/sup\u003e and 1026.94 ± 72.53 ml.hr\u003csup\u003e-1\u003c/sup\u003e.m\u003csup\u003e-2\u003c/sup\u003e, respectively.The mean plasma concentration-time profile of PCB 180 in mouse plasma after inhalation of PCB mixture at three different dose levels are presented inFigure 2, inset A, and Table 2.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePharmacokinetics profile of PCB 180 at 2.0, 4.0 and 16.0 mg.m\u003csup\u003e-3\u003c/sup\u003e in mice brain:\u003c/strong\u003eThe peak concentration of PCB 180 attained in the brain in the 2, 4 and 16 mg.m\u003csup\u003e-3\u003c/sup\u003e groups were42.04 ± 4.83, 135.49 ± 4.95 and 429.35 ± 14.37 ng.ml\u003csup\u003e-1\u003c/sup\u003e,respectively, showing a geometrically progressive trend with dose. The Tmax of this congener in brain were 2.0 ± 0.0, 2.0 ± 0.0, and 6.0 ± 0.0 hr in the threedose levels, respectively. The AUC0-t in brain were 631.12 ± 46.60, 2449.37 ± 186.79, and 8332.99 123.54, ng.hr.ml\u003csup\u003e-1\u003c/sup\u003e respectively with dose increments; theAUC\u003csub\u003e0-∞\u003c/sub\u003e for PCB 180 across the three dose levels were 915.48 ± 106.63, 3896.91 ± 367.99, and 10984.01 ± 590.37 ng.hr.ml\u003csup\u003e-1\u003c/sup\u003e, respectively. The K\u003csub\u003eel\u0026nbsp;\u003c/sub\u003ein brain were 0.03, 0.02 and 0.03 hr\u003csup\u003e-1\u003c/sup\u003e across the three graded doses of Aroclor 1232. The half-lives of PCB 180 elimination from brain were 22.17 ± 3.24, 31.43 ±3.10, and 22.73 ± 3.57 hr, while the MRT were 13.88 ± 0.21, 19.45 ± 0.64, and 18.72 ± 0.21 hr correspondingly, in the brain. The mean apparent Vd of PCB180 in the mouse brains in the 2, 4 and 16 mg.m\u003csup\u003e-\u003c/sup\u003e3 doses were 69838.36 ± 5306.26, 46934.13 ± 6867.21 and, 47593.68 ± 4968.74 ml.m\u003csup\u003e-2\u003c/sup\u003e and its clearancewere 2207.04 ± 256.46, 1033.24 ± 96.14 and 1459.71 ± 75.49 ml.hr\u003csup\u003e-1\u003c/sup\u003e.m\u003csup\u003e-2\u003c/sup\u003e.The mean brain concentration-time profile of PCB 180 in mouse brain after inhalation of PCB mixture at three different dose levels are presented in Figure 2,inset B, and Table 2 (Supplemental files 1A and 1B).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eDose dependence of plasma PCB 180 and PCB 77\u003c/u\u003e\u003c/strong\u003e:\u003c/p\u003e\n\u003cp\u003eAroclor 1232 was dosed to Swiss albino mice at three dose levels – 2.0 (low dose), 4.0 (intermediate dose) and 16.0 (high dose) mg.m\u003csup\u003e-3\u003c/sup\u003e. PCB 77Cmax in plasma was eight times higher in high dose compared to low dose level. However, this was only three times higher in intermediate dose group compared to 2.0 mg.m\u003csup\u003e-3\u003c/sup\u003e dose level and three times higher for high dose compared to 4.0 mg.m\u003csup\u003e-3\u003c/sup\u003e. In plasma, PCB 180 Cmax was six times and twelve timeshigher for intermediate and high dose groups compared to 2.0 mg.m\u003csup\u003e-3\u003c/sup\u003e dose levels. The high dose level Cmax for plasma PCB 180 was twice thanintermediate dose level.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDose-dependent translation of brain to plasma PCB 77, 180 ratios:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePCB 77 brain to plasma ratios were similar for intermediate and high dose level, which reached about 1.1 times for lower dose level. The ratios for PCB 180 were three times in the intermediate dose level compared to high dose; the ratio was 5 times higher in low dose compared to the high dose level, and 1.7times high in intermediate dose compared to low dose (Figure 3, Table 3, Supplemental files 2A and 2B).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eEffect of brain to plasma ratio of PCB 77 and PCB 180 in Swiss albino mice:\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHill model was used to quantify the Inhibitory Dose, ID-50 values for PCB 180 and PCB 77 to depress behavioral scores at the same level of brain toplasma ratios (Figure 3, and, 4). PCB 180 ID-50 was 7.645 mg.m\u003csup\u003e-3\u003c/sup\u003e, and this was higher than PCB 77 ID-50 values (7.382 mg.m\u003csup\u003e-3\u003c/sup\u003e). The slopes of lines were13.89 and 12.42, and Y-Intercepts for behavioral disruption were 38.23 and 37.16 respectively for PCB 77 and PCB 180.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eEffects of Aroclor 1232 on behavior\u003c/u\u003e\u003c/strong\u003e:\u003c/p\u003e\n\u003cp\u003eThere was a significant difference in total scores at 3 and 4 hours for all three doses. Significant difference in mean scores was also observed at 5 hours of single-6-hour exposures for 2 and 4 mg.m\u003csup\u003e-3\u003c/sup\u003e doses (p\u0026lt; 0.05), when compared to control group. The sub scores that contributed to greater than 50%deviation from the normative values, as assessed by Mann Whitney U test were distributed across three main domains – mood, autonomic activity, andmiscellaneous functions, for all dose levels. The Cmax of 2, 4, and 16 mg.m\u003csup\u003e-3\u003c/sup\u003e dose groups were achieved at 3.5 ± 1.00, 4.0 ± 0 and 4.0 ± 0 hr for PCB 77 inplasma and brain. The Tmax of PCB 180 plasma and brain at 2, 4, and 16 mg.m\u003csup\u003e-\u003c/sup\u003e3 dose levels were 2.0 ± 0, 2.0 ± 0 and 6.0 ± 0 hr. A more detailed analysis ofsubdomain score across doses revealed that grooming and restlessness (mood), urination, heart and respiratory rates, skin colour and palpebral opening(autonomic), and exploratory and group clustering behaviors (miscellaneous domain) were contributing to the statistical significance in the behavioralchanges compared to control group (Supplemental files 3A and 3B).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eBrain monoamine concentrations at Tmaxof PCB congeners after exposures to different doses of Aroclor 1232\u003c/u\u003e\u003c/strong\u003e:\u003c/p\u003e\n\u003cp\u003eThe brain concentrations of Noradrenaline at 4 hr (Cmax of PCB 77 in brain) were 3.8, 1.7 and 2.0 ng/ml across 2, 4, and 16 mg.m\u003csup\u003e-3\u003c/sup\u003edoses. Dopamine (DA)reached 4.8, 1.6 and 7.5 ng/ml in 2, 4 and 16 mg.m\u003csup\u003e-3\u003c/sup\u003edoses at 4 hr. (Tmax of PCBs in brain).At 4 hr corresponding to Tmax of PCBs in brain, the brain 5-HTlevels were 3.0, 2.6 and 4.3 ng.ml\u003csup\u003e-1\u003c/sup\u003e in 2, 4 and 16 mg.m\u003csup\u003e-\u003c/sup\u003e3doses. Brain levels of Noradrenaline and Dopamine were reduced as PCB 77 concentrationsreached maximum in brain. PCB 180 levels did not correlate with either central behavioral disruption or brain levels of all three monoamines.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eEffects of Aroclor 1232 on brain monoamine levels: Relationship with behavioral scores:\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBrain monoamine levels were estimated using HPLC-FLD in the first six hours for each dose level. Brain monoamine levels followed a linearrelationship for 5-HT, exponential relationship for NA and non-monotonic or bimodular for DA in the first six hours of exposures. These could be correlatedwith behavioral scores at Cmax time points for each dose level (Figure 5, Table 4, Table 5).\u003c/p\u003e\n\u003cp\u003eGenerally, a decline in mood was noted due to dipping of scores for grooming and restlessness sub-domains. The mean grooming scores across all days dipped at 5 hours for higher dose levels as compared to 4 and 6 hours for 2 mg.m\u003csup\u003e-3\u003c/sup\u003e dose. The trough in mean restlessness scores across all days was 3 hours for low dose and this shifted by one hour for each geometric increase in dose (Supplemental files 4A, 4B and 4C).\u003c/p\u003e\n\u003cp\u003eIn 2 mg.m\u003csup\u003e-3\u003c/sup\u003eexposed animals, the mood was generally depressed due to reduction in grooming at 3 and 4 hours, and increase in restlessness at 3, 4, and 5-hour time points. Grooming scores was back to control values at 5 hours in this dose, whereas increased restlessness did not recover even at 6 hours in this group. In the 4 mg/cu.m dose groups, grooming scores dipped at 3, 4, and 5-hour time points. But restlessness scores increased at 3, 4 and 5-hour time points. In the 16 mg.m-3dose level, the grooming reduction was maximum at 3, 4, and 5 hr after exposure and did not recover even at 6 hr in this dose level.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eRestlessness was increased 3 hr, after which it came to normal at 4 hr time point in this dose level. At 5 hr after exposure, the restlessness scores increased, which did not recover back in this dose level.\u003c/p\u003e\n\u003cp\u003eIn this study, the mean palpebral opening was below the normative value at all time points, with the maximum dip approached at 4 hours for 2 mg.m\u003csup\u003e-3\u003c/sup\u003e dose and at 5 hours for higher doses. Although the heart rate was below the normative score during all time points, there was a tendency for maximum bradycardia at 3-5 hours across all dose levels. Similarly, the respiratory rate tended to be below the normative always and to initially dip at about 2-3 hours and then recover to baseline value at 5 hours across all days and dose levels. Urination scores increased at 5 hours for 2 and 4 mg.m\u003csup\u003e-3\u0026nbsp;\u003c/sup\u003edoses across the day averages, whereas it reduced at 4 and 5 hours for the 16 mg.m\u003csup\u003e-3\u003c/sup\u003e dose.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAmong the miscellaneous domains, group clustering (social behavior) activity increased with duration of exposure as the animals tended to reduce the surface area of their fur to avoid exposure to the potentially irritating PCBs. This was inversely proportional to the exploratory activity of mice in the chamber. Exploratory activity increased at 3, 4, and 5 hr in 2, 4, and 16 mg.m\u003csup\u003e-3\u003c/sup\u003e dose levels. Social behavior increased at 3, 4, and 5 hr in 2 and 16 mg.m\u003csup\u003e-3\u0026nbsp;\u003c/sup\u003edose levels. However, in the 4 mg.m\u003csup\u003e-3\u003c/sup\u003e dose level, the social behavior scores were normal at 3 hr, then it decreased at 4 hr, after which it again recovered back to normal at 5 hr (Table 5).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eRelationship of sub-domain behavior scores across doses:\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe total behavioral subdomain scores across each time point compared pair wise during single 6-hour exposure to Aroclor 1232 exhibited dose dependence at 2.0, 4.0 and 16.0 mg.m\u003csup\u003e-3\u003c/sup\u003e dose levels (F ratio = 18.41, p \u0026lt; 0.0001). The Pearson Correlation coefficients were close to 1.0 for most of the pairwise comparisons. One-way Repeated Measures ANOVA of each correlation coefficients showed that the significance was due to inter-dose variation than inter-individual variability (p = 0.0077).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eDose dependence of sub-domain scores at Tmax\u003c/u\u003e\u003c/strong\u003e:\u003c/p\u003e\n\u003cp\u003eLikewise, the behavior scores were statistically significant at Tmax of PCB 77 and PCB 180 (3-5 hours) with p value 0.0011. Friedman test of the correlation coefficients at Tmax was significant across doses (Friedman statistic = 31.04). There was more of behavioral depression than stimulatoryeffects of PCBs at Tmax.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cu\u003eContribution of sub-domains to dose-dependent behavioral effects of PCBs at Tmax:\u003c/u\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMood, which composed of five sub-domains namely grooming, restlessness, vocalization, irritability, and fearfulness, was generally depressedacross all doses of exposures. This was generally ascribed to depression in grooming behavior on Irwin scale across all three doses, although restlessnessseemed to increase dose-dependently. Concentrations of PCB 77 and PCB 180 in lung, brain and plasma significantly correlated negatively with mooddepression at 6 hours in each dose level (-0.75 \u0026lt;Pearson ‘r’\u0026lt; -0.85, Figure 6). Grooming depression was maximum at 4 hours and Restlessness wasmaximally increased at 4 hours across all doses of exposure (Figure 7).\u003c/p\u003e\n\u003cp\u003eOf the eleven autonomic system functions including palpebral opening, writhing response, pupil size, exophthalmos, pupil size, urination, salivation, hypothermia, skin colour, heart rate and respiratory rate, palpebral opening, urination and skin colour increased at 5 hours of exposure, especially at 4.0 and16.0 mg.m\u003csup\u003e-3\u003c/sup\u003e doses. However, heart rate and respiration, reduced at 3 hours maximally across all three doses.\u003c/p\u003e\n\u003cp\u003eOut of the three miscellaneous domains (social aggregation, hostility, and exploratory activity), social conglomeration increased at 3-5 hours acrossall three doses, and exploratory activities increased at 3-5 hours across 2.0 and 16.0 mg.m\u003csup\u003e-3\u003c/sup\u003e dosing groups.Generally, the peak of central behavioral disruption followed the Tmax of PCB 77, whereas PCB 180 Cmax contributed more to the peripheral effects onbehavior (Table 4, and, Table 5, Supplemental files 1A, 1B, and 5).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe Inhalation dosing of mice with Aroclor 1232 in the whole-body inhalation chamber was successful in producing significant plasma and brainconcentrations of the two representative PCB congeners. The pharmacokinetic parameters of these two congeners in the two matrices were hence derivedwith high degree of correlation across the dose levels. Plasma peak concentrations of both congeners across all the three doses were generally one-andhalftimes greater than the brain levels. The time to peak absorption of both the congeners were nearly two times longer for PCB 77 than PCB 180, and this was generally independent of dose and tissue matrix. Higher (nearly twice) AUCs were obtained for PCB 77 in both the matrices than PCB 180 generally across the three dose levels. The plasma AUC was generally higher than the brain AUC for both the congeners across the three dose levels, with dose proportionality.\u003c/p\u003e \u003cp\u003ePCBs are known to possess high orders of K\u003csub\u003eow\u003c/sub\u003e providing them with high lipophilicity and a capacitance to traverse across biological membranes [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. PCBs can travel across biological barriers by both passive diffusion and active transport mechanisms. Hence the individual PCB congeners would be expected to cumulate and have longer biological half-lives. However, as is apparent from the acute inhalation study of Aroclor 1232, dosing PCB mixtures can impart only intermediately long half-lives (in hours) and MRTs for certain key toxic congeners like PCB 77 and even the hepta-CB, PCB 180, in plasma and brain tissue as well. This may also explain the short elimination rate constants, which are inversely correlated with log K\u003csub\u003eow\u003c/sub\u003e, and hence lipophilicity of congeners [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Though tetra-CB congeners like PCB 77 do not accumulate in aquatic fishes and other environmental reservoirs [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], certain high chlorinated congeners can attain significantly toxic concentrations and large AUCs in mouse and human matrices, as can be deduced from the large peak plasma and brain concentrations of PCB 180 and PCB 77, especially in higher doses. Such results have similarly been obtained with other investigations [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] where PCB 77 concentrations in different areas of the brain like the striatal cortex have been generically less compared to PCB 180 concentrations, after feeding rats with Aroclor 1254 in drinking water and chow. Although a dynamic equilibrium exists among the final tissue concentrations of homologs of PCBs as suggested by studies elsewhere [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] the concentrations of certain toxic congeners can significantly vary in same organism in different tissues [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. In our study, both congeners generally have large volumes of distribution and clearance rates from both plasma and brain, which are dose-dependent and not time-of-exposure-dependent.\u003c/p\u003e \u003cp\u003eMany PCBs especially higher chlorinated PCBs like PCB 180 can have active metabolites in plasma and other tissues like methoxy, sulphated and hydroxyl metabolites, which could shorten the elimination rate constants of such congeners in-vivo [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Hence, a change in rate of elimination of a congener from a tissue matrix with increments in dose usually depends on two factors \u0026ndash; the rate of metabolism to active metabolites, and the rate of excretion versus distribution constants. Since PCB 180 is more lipophilic chlorinated and may have a greater number of potent active metabolites, its volumes of distribution and elimination half-lives change more drastically with dose increments, than PCB 77. Finally, for most of the pharmacokinetic parameters calculated for both congeners in this study, the variations imparted by the inhalation route is less, and the dose-dependent nature of their linear pharmacokinetics in brain and plasma is of a geometrically graded nature.\u003c/p\u003e \u003cp\u003eSince PCB 77 is more of a dioxin like PCB with ability to bind to AhRs and cause neuroendocrine disruption more than PCB 180, this study confirms the hypothesis that environmental reminisce of such PCBs, though seemingly low chlorinated can lead to monoamine and other neurotransmitter alterations in the brain even in rodents, via entry into the biological systems by inhalation routes. Further, such alterations could theoretically lead to neurobehavioral disruptions and neurological disorders if they accumulate in the brain as life progresses.\u003c/p\u003e \u003cp\u003eLipophilicity of PCB congeners depends on number of chlorine atoms on the biphenyl nucleus and absorption of PCBs onto biological membranes in turn depends on lipophilicity and steric hindrance caused by position of chlorine on biphenyl congener [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Ultimately absorption of PCBs in biological systems is influenced by the physical and stereo-chemical properties, and structure-activity and structure-metabolism relationships [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. PCBs are known to sequester into tissues richly supplied by blood flow like intestine wall and this occurs by passive diffusion [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Since PCBs are known to get absorbed via oral, dermal, and inhalation routes, the main routes of excretion include, obviously, fecal, cutaneous, and respiratory air.\u003c/p\u003e \u003cp\u003ePCB congeners are known to accumulate in adipocytes, irrespective of dose and species, apart from blood - hence, their distribution in vivo is determined by the blood flow and lipophilicity directly [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].Generally, ortho-substituted PCBs have higher partition coefficients as compared to non-ortho and dioxin-like PCBs which possess all forms of chlorine substitutions [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. PCB 77 being a dioxin-like highly lipid soluble PCB sequesters to brain but cannot be removed easily due to tissue sequestration in brain tissues and cause thyroid and neuroendocrine disruption [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. However, PCBs are known to disrupt Blood Brain Barrier and enter the brain easily on high doses [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Hepatic binding to cytochrome enzymes also determines their higher hepatic concentrations. PCBs are known to induce the metabolism of other xenobiotics including PCBs themselves. Further fate and accumulation of PCBs are determined by what fraction of the administered congener is metabolized and how fast this process happens. This depends on chemistry of each congener [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Coplanar PCBs which have favorable spatial arrangement for hindering metabolism, and mono-ortho substituted PCBs, do not get metabolized easily [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Unsubstituted chlorine at para-position changes the ability of the congener to form toxic arene metabolites easily causing free radical damage. [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Non-substituted PCBs at ortho-meta positions and possession of higher number of chlorine atoms like PCB 180, makes the congener to cumulate, especially after higher or repeated dosing [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eElimination kinetics of PCB congeners vary between tissue compartments and the dose-exposure protocol. For example, the half-life of 3,3',4,4'-TCB was1.07\u0026ndash;2.60 days, since this congener sequesters in adipocytes, liver and thymocytes [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Congeners which do not tissue-cumulate, like 2,2',5,5'-TCB, havehalf-lives from 1.64\u0026ndash;2.90 days. Since their concentrations were similar despite this difference in tissue sequestration, the property that decides eliminationkinetics is therefore potency of congener, apart from chirality of stereochemistry [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. PCB 77 being highly more potent than PCB 180 in its toxic effects,makes it selectively cumulate in brain and produce behavioral depression and shorter plasma elimination half-lives [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe key results for behavioral effects of the PCB mixture include a decline in mood, autonomic and miscellaneous domain sub-scores, in a dose dependentand time dependent manner at each dose level, being contributed significantly by alteration in certain sub domains. The decline in grooming in this study at 2\u0026ndash;4 hours after exposure initiation hence is not due to introduction of mice to novel environment of chamber and can be ascribed to be influenced only bythe PCB induced neurotransmitter changes in brain. The negative chronotropic effects at 2 hours of exposure to PCBs may be due to cardiac, reflex, and central depression. This is also the time when PCBs are about to redistribute from blood to the brain after being absorbed from the alveoli. Respiratory rate reduced at 4 hours of daily exposures especially in high dose exposures. Dose-responses with monoamines followed linear (5-HT), non-monotonic (DA) and exponential (NA) relationships till 6 hours of exposure.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eGenerally, DA reduced at Tmax for each dose level; 5-HT increased exponentially as Tmax approached for each dose level; and NA followed U-shaped dose response, the dip being correlated at Cmax. PCBs are known to affect monoamine levels in neurons inside and outside the Central Nervous System, via multiple mechanisms, including inhibition of VMAT-2 and reduction of dopamine levels in synapse [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. In their ability to alter dopamine levels in brain and cause depression in humans, non-coplanar PCBs are more potent, as in this study wherein the decrease in dopamine and mood were maximum at PCB 77 Tmax. The probable mechanisms of these behavioral effects of PCB 77 inhalation on mood are outlined in Fig.\u0026nbsp;7, which may be relevant in causing neuropsychiatric illnesses (e.g. major depression, autism, OCD) in human patients. In conclusion, this study proves that PCBs can alter behavior and monoamine levels via inhalation route.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003e5-HT = 5-hydroxytrytamine; Acetonitrile (ACN); ANOVA= Analysis of Variance; AUC0-t = Area-Under-Plasma-Concentration-time-Curve from time = 0 to time = t; AUC0-\u0026infin; = Area-Under-Plasma- Concentration-time-Curve from time = 0 to time = infinity; CB = Chloro-biphenyl; CPCSEA = Committee for the Purpose ofControl and Supervision of Experimentation in Animals; Cmax = Maximum Concentration; CL = Clearance; DA = Dopamine; DAD = Diode Array Detector;EDTA = Ethylene Diamine Tetra Acetate; FLD = Fluorescence detector; HPLC = High-Performance Liquid Chromatography; IAEC = Institutional Animal EthicsCommittee; ID-50 = Inhibitory Dose-50%; IS = Internal Standard; K\u003csub\u003eel\u003c/sub\u003e = Elimination rate constant; K\u003csub\u003eow\u0026nbsp;\u003c/sub\u003e= Log (Octanol-water) partition coefficient; OECD =Organization for Economic Cooperation \u0026amp; Development; MRT = Mean Residence Time; NA = Noradrenaline; PCBs = Polychlorinated Biphenyls; PBS =Phosphate Buffered Saline; QC = Quality Control; RM-ANOVA = Repeated Measures-Analysis of Variance; t1/2 = Half-life; TBME = tertiary-Butyl Methyl Ether;Tmax = Maximum Time; UPLC = Ultra-high Performance Liquid Chromatography; UV = Ultraviolet; Vd = Volume of Distribution.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval \u003c/strong\u003e- The study was carried out after PSG-IAEC approval from a CPCSEA-approved Animal Facility. The study was conducted in accordance with CPCSEA guidelines for laboratory animal facility, 2005.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate\u003c/strong\u003e \u0026ndash; Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e - It has been approved by all the authors listed as per ICJME criteria.\u003c/p\u003e\n\u003cp\u003eAvailability of data and materials - The manuscript includes data which is linked to the study results. This data collection can be made available on reasonable and justified request and shall be bound by the publishers\u0026rsquo; data management policy for submitting manuscripts involving original research. This data policy does not breach any confidentiality and is based sound ethical compliance. Infact, the supplementary files have been attached for initial review and plausible publication, including review thereafter, to augment and enhance the clarity, reproducibility, and authenticity of the results discussed, with prior approval of all the authors. Preprint available at DOI: https://doi.org/10.21203/rs.3.rs-1425455/v1\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests \u003c/strong\u003e- The authors declare no conflicts of interests during the conception and inception of the paper, including any non-financial and monetary ones. All the authors have read and approved the manuscript, and all data were generated in-house and that no paper mill was used.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding \u003c/strong\u003e- The authors declare that the study was self-funded with no other financial funding contributions / drawing from outside by the authors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions -\u003c/strong\u003e SSK contributed to manuscript editing in addition to data analysis and visualization and chromatography (new methods). NR contributed to study inception, design, data analysis, manuscript writing and editing and data analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e - The authors would like to acknowledge the facilities given by PSG Institute of Medical Sciences \u0026amp; Research, Coimbatore for the success of the work. The authors acknowledge the contributions of the former Dean and Professor of Pharmacology and PSG-CMMT, Dr. Ramalingam Sankaran (formerly), for his administrative and logistic facilities provision, as the Guide of the Part time PhD program of one of the authors.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eStockholm Convention - Home page. http://www.pops.int/default.aspx. Accessed 25 Jan 2022\u003c/li\u003e\n\u003cli\u003eHealth UD of, Services H (1999) Agency for Toxic Substances and Disease Registry-ATSDR.\u003c/li\u003e\n\u003cli\u003eEriksson P, Fredriksson A (1996) Developmental neurotoxicity of four ortho-substituted polychlorinated biphenyls in the neonatal mouse. Environ ToxicolPharmacol 1:155\u0026ndash;165. https://doi.org/10.1016/1382-6689(96)00015-4\u003c/li\u003e\n\u003cli\u003eSeegal RF (1996) Epidemiological and laboratory evidence of PCB-induced neurotoxicity. Crit Rev Toxicol 26:709\u0026ndash;737. https://doi.org/10.3109/10408449609037481\u003c/li\u003e\n\u003cli\u003eSimon T, Britt JK, James RC (2007) Development of a neurotoxic equivalence scheme of relative potency for assessing the risk of PCB mixtures. RegulToxicolPharmacol 48:148\u0026ndash;170. https://doi.org/10.1016/j.yrtph.2007.03.005\u003c/li\u003e\n\u003cli\u003eLouis C, Covaci A, Crocker DE, Debier C (2016) Lipophilicity of PCBs and fatty acids determines their mobilisation from blubber of weaned northern elephant seal pups. Sci Total Environ 541:599\u0026ndash;602. https://doi.org/10.1016/j.scitotenv.2015.09.094\u003c/li\u003e\n\u003cli\u003eIrwin S (1968) Comprehensive observational assessment: Ia. A systematic, quantitative procedure for assessing the behavioral and physiologic state of the mouse. Psychopharmacologia 13:222\u0026ndash;257. https://doi.org/10.1007/BF00401402\u003c/li\u003e\n\u003cli\u003eCommittee for the Purpose of, Animals Supervision of Experiments on Animals (2003) CPCSEA Guidelines for laboratory animal facility. Indian J Pharmacol 35: \u003c/li\u003e\n\u003cli\u003eOECD (2009) OECD guidance document on acute inhalation toxicity testing. The Environment, Health and Safety Division, Organization for Economic Cooperation \u0026amp; Development\u003c/li\u003e\n\u003cli\u003eNarayanan R, Muthusamy S, Lakshmi SV, et al (2021) Design and development of whole-body rodent inhalation chamber for exposure to Aroclor 1232 in Swiss albino mice. Int J Environ Sci Technol. https://doi.org/10.1007/s13762-021-03515-8\u003c/li\u003e\n\u003cli\u003eRamanujam N, Sivaselvakumar M, Ramalingam S (2017) Fast and parallel determination of PCB 77 and PCB 180 in plasma using ultra performance liquid chromatography with diode array detection: A pharmacokinetic study in Swiss albino mouse. Biomed Chromatogr31:. https://doi.org/10.1002/bmc.4000\u003c/li\u003e\n\u003cli\u003eWang H, Walaszczyk EJ, Li K, et al (2012) High-performance liquid chromatography with fluorescence detection and ultra-performance liquid chromatography with electrospray tandem mass spectrometry method for the determination of indoleamine neurotransmitters and their metabolites in sea lamprey plasma. Anal Chim Acta 721:147\u0026ndash;153. https://doi.org/10.1016/j.aca.2012.01.025\u003c/li\u003e\n\u003cli\u003eGrandjean P, Budtz-J\u0026oslash;rgensen E, Barr DB, et al (2008) Elimination half-lives of polychlorinated biphenyl congeners in children. Environ Sci Technol 42:6991\u0026ndash;6996. https://doi.org/10.1021/es800778q\u003c/li\u003e\n\u003cli\u003eCarpenter DO (2015) Exposure to and health effects of volatile PCBs. Rev Environ Health 30:81\u0026ndash;92. https://doi.org/10.1515/reveh-2014-0074\u003c/li\u003e\n\u003cli\u003eSeegal RF, Okoniewski RJ, Brosch KO, Bemis JC (2002) Polychlorinated biphenyls alter extraneuronal but not tissue dopamine concentrations in adult rat striatum: an in vivo microdialysis study. Environ Health Perspect 110:1113\u0026ndash;1117. https://doi.org/10.1289/ehp.021101113\u003c/li\u003e\n\u003cli\u003eMatthews HB, Dedrick RL (1984) Pharmacokinetics of PCBs. Annu Rev PharmacolToxicol 24:85\u0026ndash;103. https://doi.org/10.1146/annurev.pa.24.040184.000505\u003c/li\u003e\n\u003cli\u003eBerninger JP, Tillitt DE (2019) Polychlorinated biphenyl tissue-concentration thresholds for survival, growth, and reproduction in fish. Environ Toxicol Chem 38:712\u0026ndash;736. https://doi.org/10.1002/etc.4335\u003c/li\u003e\n\u003cli\u003eGrimm FA, Hu D, Kania-Korwel I, et al (2015) Metabolism and metabolites of polychlorinated biphenyls. Crit Rev Toxicol 45:245\u0026ndash;272. https://doi.org/10.3109/10408444.2014.999365\u003c/li\u003e\n\u003cli\u003eClevenger MA, Roberts SM, Lattin DL, et al (1989) The pharmacokinetics of 2,2\u0026rsquo;,5,5\u0026rsquo;-tetrachlorobiphenyl and 3,3\u0026rsquo;,4,4\u0026rsquo;-tetrachlorobiphenyl and its relationship to toxicity. Toxicol Appl Pharmacol 100:315\u0026ndash;327. https://doi.org/10.1016/0041-008x(89)90317-7\u003c/li\u003e\n\u003cli\u003eLotti M (2003) Pharmacokinetics and blood levels of polychlorinated biphenyls. Toxicol Rev 22:203\u0026ndash;215. https://doi.org/10.2165/00139709-200322040-00003\u003c/li\u003e\n\u003cli\u003eCorrigan FM, Murray L, Wyatt CL, Shore RF (1998) Diorthosubstituted polychlorinated biphenyls in caudate nucleus in Parkinson\u0026rsquo;s disease. Exp Neurol 150:339\u0026ndash;342. https://doi.org/10.1006/exnr.1998.6776\u003c/li\u003e\n\u003cli\u003eParham FM, Portier CJ (1998) Using structural information to create physiologically based pharmacokinetic models for all polychlorinated biphenyls. II. Rates of metabolism. Toxicol Appl Pharmacol 151:110\u0026ndash;116. https://doi.org/10.1006/taap.1998.8441\u003c/li\u003e\n\u003cli\u003eParham FM, Kohn MC, Matthews HB, et al (1997) Using structural information to create physiologically based pharmacokinetic models for all polychlorinated biphenyls. Toxicol Appl Pharmacol 144:340\u0026ndash;347. https://doi.org/10.1006/taap.1997.8139\u003c/li\u003e\n\u003cli\u003eKodavanti PR, Ward TR, Derr-Yellin EC, et al (1998) Congener-specific distribution of polychlorinated biphenyls in brain regions, blood, liver, and fat of adult rats following repeated exposure to Aroclor 1254. Toxicol Appl Pharmacol 153:199\u0026ndash;210. https://doi.org/10.1006/taap.1998.8534\u003c/li\u003e\n\u003cli\u003eOberg M, Sj\u0026ouml;din A, Casabona H, et al (2002) Tissue distribution and half-lives of individual polychlorinated biphenyls and serum levels of 4-hydroxy-2,3,3\u0026rsquo;,4\u0026rsquo;,5-pentachlorobiphenyl in the rat. Toxicol Sci 70:171\u0026ndash;182. https://doi.org/10.1093/toxsci/70.2.171\u003c/li\u003e\n\u003cli\u003eBeck V, Roelens SA, Darras VM (2006) Exposure to PCB 77 induces tissue-dependent changes in iodothyronine deiodinase activity patterns in the embryonic chicken. Gen Comp Endocrinol 148:327\u0026ndash;335. https://doi.org/10.1016/j.ygcen.2006.04.003\u003c/li\u003e\n\u003cli\u003eSimmons SL, Cummings JA, Clemens LG, Nunez AA (2005) Exposure to PCB 77 affects the maternal behavior of rats. PhysiolBehav 84:81\u0026ndash;86. https://doi.org/10.1016/j.physbeh.2004.10.022\u003c/li\u003e\n\u003cli\u003eJandacek RJ, Rider T, Keller ER, Tso P (2010) The effect of olestra on the absorption, excretion and storage of 2,2\u0026rsquo;,5,5\u0026rsquo; tetrachlorobiphenyl; 3,3\u0026rsquo;,4,4\u0026rsquo;tetrachlorobiphenyl; and perfluorooctanoic acid. Environ Int 36:880\u0026ndash;883. https://doi.org/10.1016/j.envint.2009.06.010\u003c/li\u003e\n\u003cli\u003eSeelbach M, Chen L, Powell A, et al (2010) Polychlorinated biphenyls disrupt blood-brain barrier integrity and promote brain metastasis formation. Environ Health Perspect 118:479\u0026ndash;484. https://doi.org/10.1289/ehp.0901334\u003c/li\u003e\n\u003cli\u003eQuinete N, Esser A, Kraus T, Schettgen T (2017) PCB 28 metabolites elimination kinetics in human plasma on a real case scenario: Study of hydroxylated polychlorinated biphenyl (OH-PCB) metabolites of PCB 28 in a highly exposed German Cohort. Toxicol Lett 276:100\u0026ndash;107. https://doi.org/10.1016/j.toxlet.2017.05.025\u003c/li\u003e\n\u003cli\u003eKania-Korwel I, Lehmler H-J (2016) Chiral polychlorinated biphenyls: absorption, metabolism and excretion--a review. Environ Sci Pollut Res Int 23:2042\u0026ndash;2057. https://doi.org/10.1007/s11356-015-4150-2\u003c/li\u003e\n\u003cli\u003eDhakal K, Gadupudi GS, Lehmler H-J, et al (2018) Sources and toxicities of phenolic polychlorinated biphenyls (OH-PCBs). Environ Sci Pollut Res Int 25:16277\u0026ndash;16290. https://doi.org/10.1007/s11356-017-9694-x\u003c/li\u003e\n\u003cli\u003eHaraguchi K, Koga N, Kato Y (2005) Comparative metabolism of polychlorinated biphenyls and tissue distribution of persistent metabolites in rats, hamsters, and Guinea pigs. Drug MetabDispos 33:373\u0026ndash;380. https://doi.org/10.1124/dmd.104.002444\u003c/li\u003e\n\u003cli\u003eDanis B, Cotret O, Teyssi\u0026eacute; JL, et al (2004) Coplanar PCB 77 uptake kinetics in the sea star Asterias rubens and subsequent effects on reactive oxygen species (ROS) production and levels of cytochrome P450 immunopositive proteins (CYP1A-IPP). Marine Ecology Progress Series 279:117\u0026ndash;128. https://doi.org/10.3354/meps279117\u003c/li\u003e\n\u003cli\u003eKania-Korwel I, Lehmler H-J (2016) Toxicokinetics of chiral polychlorinated biphenyls across different species--a review. Environ Sci Pollut Res Int 23:2058\u0026ndash;2080. https://doi.org/10.1007/s11356-015-4383-0\u003c/li\u003e\n\u003cli\u003eTan Y, Song R, Lawrence D, Carpenter DO (2004) Ortho-substituted but not coplanar PCBs rapidly kill cerebellar granule cells. Toxicol Sci 79:147\u0026ndash;156. https://doi.org/10.1093/toxsci/kfh108\u003c/li\u003e\n\u003cli\u003eViluksela M, Heikkinen P, van der Ven LTM, et al (2014) Toxicological profile of ultrapure 2,2\u0026rsquo;,3,4,4\u0026rsquo;,5,5\u0026rsquo;-heptachlorbiphenyl (PCB 180) in adult rats. PLoS ONE 9:e104639. https://doi.org/10.1371/journal.pone.0104639\u003c/li\u003e\n\u003cli\u003eBemis JC, Seegal RF (2004) PCB-induced inhibition of the vesicular monoamine transporter predicts reductions in synaptosomal dopamine content. Toxicol Sci 80:288\u0026ndash;295. https://doi.org/10.1093/toxsci/kfh153\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1:\u0026nbsp;\u003c/strong\u003eThe Mean Pharmacokinetic Parameters of PCB 77 in Plasma and Brain after inhalation of PCBs at 2, 4 and 16 mg/m\u003csup\u003e3\u003c/sup\u003e dose\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"106%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.131313131313131%\" rowspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003ePharmacokinetic Parameters\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"86.86868686868686%\" colspan=\"6\"\u003e\n \u003cp\u003e\u003cstrong\u003ePCB 77\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"50%\" colspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003ePlasma\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"50%\" colspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003eBrain\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.0 mg/cu.m (mean\u0026plusmn;SD, N=4)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4.0 mg/cu.m (mean\u0026plusmn;SD, N=4)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e\u003cstrong\u003e16.0 mg/cu.m (mean\u0026plusmn;SD, N=4)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.0 mg/cu.m (mean\u0026plusmn;SD, N=4)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4.0 mg/cu.m (mean\u0026plusmn;SD, N=4)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\"\u003e\n \u003cp\u003e\u003cstrong\u003e16.0 mg/cu.m (mean\u0026plusmn;SD, N=4)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003csub\u003emax\u003c/sub\u003e(ng/mL)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e88.32\u0026plusmn;8.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e245.62\u0026plusmn;39.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e727.40\u0026plusmn;42.52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e69.43\u0026plusmn;10.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e175.13\u0026plusmn;6.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e526.21\u0026plusmn;28.63\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eT\u003csub\u003emax\u003c/sub\u003e(hr)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e3.50\u0026plusmn;1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e4.00\u0026plusmn;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e4.00\u0026plusmn;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e3.50\u0026plusmn;1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e4.00\u0026plusmn;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e2.00\u0026plusmn;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eAUC\u003csub\u003e0-t\u0026nbsp;\u003c/sub\u003e(ng.hr/mL)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e1321.19\u0026plusmn;55.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e2749.52\u0026plusmn;392.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e7565.21\u0026plusmn;148.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e1254.97\u0026plusmn;57.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e2366.40\u0026plusmn;142.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e6798.93\u0026plusmn;340.47\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eAUC\u003csub\u003e0-\u0026infin; (\u003c/sub\u003eng.hr/mL)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e1626.02\u0026plusmn;64.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e2957.28\u0026plusmn;413.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e8013.39\u0026plusmn;263.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e1469.62\u0026plusmn;161.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e2581.12\u0026plusmn;136.46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e7100.56\u0026plusmn;413.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eT\u003csub\u003e1/2\u003c/sub\u003e (hr)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e20.17\u0026plusmn;1.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e12.72\u0026plusmn;0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e11.17\u0026plusmn;1.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e16.20\u0026plusmn;4.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e14.20\u0026plusmn;0.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e10.82\u0026plusmn;1.29\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMRT\u003csub\u003elast\u003c/sub\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e16.51\u0026plusmn;0.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e14.31\u0026plusmn;0.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e14.29\u0026plusmn;0.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e17.07\u0026plusmn;1.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e15.05\u0026plusmn;0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e14.39\u0026plusmn;0.30\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eK\u003csub\u003eel\u0026nbsp;\u003c/sub\u003e(hr\u003csup\u003e-1\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e0.03\u0026plusmn; 0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e0.06\u0026plusmn;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e0.06\u0026plusmn;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e0.05\u0026plusmn;0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e0.05\u0026plusmn;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e0.07\u0026plusmn;0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eVd (ml.m\u003csup\u003e-2\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e35858.25\u0026plusmn;3073.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e25209.01\u0026plusmn;3835.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e32099.78\u0026plusmn;3593.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e31481.64\u0026plusmn;5927.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e31856.13\u0026plusmn;3012.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e35070.00\u0026plusmn;2110.32\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eCL (ml.hr\u003csup\u003e-1\u003c/sup\u003em\u003csup\u003e-2\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e1231.46\u0026plusmn;49.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e1375.07\u0026plusmn;215.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e1998.28\u0026plusmn;65.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e1373.32\u0026plusmn;150.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e1553.12\u0026plusmn;85.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.43298969072165%\"\u003e\n \u003cp\u003e2258.90\u0026plusmn;127.06\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2:\u0026nbsp;\u003c/strong\u003eTabulation of mean monoamine concentrations at T\u003csub\u003emax\u003c/sub\u003e after exposure to different doses of Aroclor 1232\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"851\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.336075205640423%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5-HT (ng/ml)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.554641598119858%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e2 mg/cu.m\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.554641598119858%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e4 mg/cu.m\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.554641598119858%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e16 mg/cu.m\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\u0026nbsp;\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003eSD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003eSD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003eSD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2 hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.07\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.07\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.11\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.13\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.08\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.23\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4 hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.04\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.27\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.62\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.22\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4.32\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.47\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e6 hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4.33\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.67\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.29\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.23\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4.36\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.18\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.336075205640423%\"\u003e\n \u003cp\u003e\u003cstrong\u003eDA (ng/ml)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.554641598119858%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e2 mg/cu.m\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.554641598119858%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e4 mg/cu.m\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.554641598119858%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e16 mg/cu.m\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2 hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.99\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.54\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.75\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.45\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.39\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.04\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4 hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.42\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.64\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.83\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.13\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.73\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.48\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e6 hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5.15\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.85\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4.91\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.51\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5.07\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.46\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.336075205640423%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNA (ng/ml)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.554641598119858%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e2 mg/cu.m\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.554641598119858%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e4 mg/cu.m\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"28.554641598119858%\" colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e16 mg/cu.m\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2 hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.24\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.08\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.27\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.16\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.23\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.15\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4 hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.88\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.25\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.85\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.11\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.09\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e6 hour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5.28\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.67\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5.13\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.35\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.15\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.15\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003eTable 3: Behavioral effects of 4 mg/cu.m dose level of Aroclor 1232 at PCB 77 T\u003csub\u003emax\u0026nbsp;\u003c/sub\u003ein brain.\u003c/p\u003e\n\u003ctable\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:192.2pt;border:solid windowtext 1.0pt;padding:0in 5.4pt 0in 5.4pt;height:8.2pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003eBEHAVIOURAL DOMAIN\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:239.9pt;border:solid windowtext 1.0pt;border-left: none;padding:0in 5.4pt 0in 5.4pt;height:8.2pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003eBEHAVIOURAL SUB-DOMAIN\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:266.3pt;border:solid windowtext 1.0pt;border-left: none;padding:0in 5.4pt 0in 5.4pt;height:8.2pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003eEFFECTS OF PCB 77 AT 4 Hrs. (4 mg/cu.m of AROCLOR 1232)\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eAWARENESS\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(192, 0, 0);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;color:#C00000;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eALERTNESS\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(192, 0, 0);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eVISUAL PLACING\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003ePASSIVITY\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eSTEREOTYPY\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eMOOD\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(192, 0, 0);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eGROOMING\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: red;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eVOCALIZATION\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eRESTLESSNESS\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: red;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eIRRITABILITY / AGGRESSION\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(255, 192, 0);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eFEARFULNESS\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eMOTOR ACTIVITY\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 112, 192);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eREACTIVITY TO ENVIRONMENT\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: red;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eSPONTANEOUS ACTIVITY\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: red;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eTOUCH RESPONSE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003ePAIN RESPONSE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eCNS EXCITATION\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eSTARTLE RESPONSE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eSTRAUB TAIL\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eTREMORS\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eTWITCHES\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eCONVULSIONS\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003ePOSTURE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eBODY POSTURE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eLIMB POSITION\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eMOTOR INCOORDINATION\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eSTAGGERING GAIT\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eABDOMINAL GAIT\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eRIGHTING REFLEX\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eMUSCLE TONE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eLIMB TONE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eGRIP STRENGTH\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eBODY SAG\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eBODY TONE\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eABDOMINAL TONE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eREFLEXES\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003ePINNA\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eCORNEAL\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eIFR (Ipsilateral flexor Reflex)\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eAUTONOMIC\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: yellow;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eWRITHING\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003ePUPIL SIZE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(146, 208, 80);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003ePALPEBRAL OPENING\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(146, 208, 80);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eEXOPHTHALMOS\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(146, 208, 80);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eURINATION\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: red;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eSALIVATION\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003ePILOERECTION\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eHYPOTHERMIA\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 240);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eSKIN COLOUR\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: yellow;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eHEART RATE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(192, 0, 0);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eRESPIRATORY RATE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(192, 0, 0);padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eMISCELLANEOUS\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(192, 0, 0);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eEXPLORATORY ACTIVITY\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: red;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eHOSTILITY\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: red;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eSOCIAL BEHAVIOR\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: red;padding: 0in 5.4pt;height: 8.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eLETHALITY / DEAD\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eACUTE\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 112, 192);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 192.2pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 239.9pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Times New Roman\",serif;'\u003eDELAYED\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 266.3pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 112, 192);padding: 0in 5.4pt;height: 8.2pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:right;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family: \"Times New Roman\",serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;'\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;'\u003e\u003cem\u003e\u003cu\u003eColour-coding:\u003c/u\u003e\u003c/em\u003e\u003c/p\u003e\n\u003ctable\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 45.55pt;border: 1pt solid windowtext;background: rgb(0, 112, 192);padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83.95pt;border-top: 1pt solid windowtext;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-image: initial;border-left: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003eNo change absolutely in all animals\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 45.55pt;border-top: 1pt solid windowtext;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-image: initial;border-left: none;background: yellow;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 67.05pt;border-top: 1pt solid windowtext;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-image: initial;border-left: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003eCNS Modulatory change in all animals\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 45.55pt;border-top: 1pt solid windowtext;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-image: initial;border-left: none;background: red;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65.45pt;border-top: 1pt solid windowtext;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-image: initial;border-left: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003eCNS depression in 50% animals\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 45.55pt;border-top: 1pt solid windowtext;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-image: initial;border-left: none;background: rgb(146, 208, 80);padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52.15pt;border-top: 1pt solid windowtext;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-image: initial;border-left: none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003eCNS excitation in 50% animals\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 45.55pt;border-right: 1pt solid windowtext;border-bottom: 1pt solid windowtext;border-left: 1pt solid windowtext;border-image: initial;border-top: none;background: rgb(0, 176, 240);padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83.95pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003eMild change in 50% of animals\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 45.55pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(255, 192, 0);padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 67.05pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003eCNS modulation in 50% animals\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 45.55pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(192, 0, 0);padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65.45pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003eCNS depression in all animals\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 45.55pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;background: rgb(0, 176, 80);padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003e\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 52.15pt;border-top: none;border-left: none;border-bottom: 1pt solid windowtext;border-right: 1pt solid windowtext;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style=\"font-size:13px;\"\u003eCNS excitation in all animals\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Polychlorinated Biphenyl, Aroclor 1232, Inhalation toxicity, Behavior, Irwin scale, Monoamines, Mood","lastPublishedDoi":"10.21203/rs.3.rs-4715900/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4715900/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIntroduction: Polychlorinated Biphenyls pose hazards to humans as environmental and occupational workplace pollutants, despite which, though they are regulated in India, their use is not yet banned. Hence, the pharmacokinetics and behavioral effects of PCBs after inhalational exposures of Aroclor 1232 in Swiss albino mice were assessed in this study, with an objective to correlate these effects with monoamine levels in brain and extrapolate them to rodent and human risk assessments.\u003c/p\u003e \u003cp\u003eMethods: A whole body inhalational chamber has been utilized to expose mice to different doses of Aroclor 1232 for single six-hour durations. The plasma and brain samples were used to estimate the corresponding concentration-time profiles of two representative congeners of PCBs (PCB 77 and PCB 180) and calculate their pharmacokinetic parameters. These variables were then correlated with mouse behavior using Irwin scale, and monoamine levels at Tmax.\u003c/p\u003e \u003cp\u003eResults \u0026amp; Discussion: A definite dose-dependence of PCB 77 and PCB 180 kinetic parameters was identified in plasma and brain. PCB 77 exhibited more accumulation in brain than plasma, whereas PCB 180 accumulated more in plasma than brain. Mood, autonomic scores, and miscellaneous sub domains were significantly different from control group scores in these domains. The changes in the mood and miscellaneous domains on the Irwin scale could be ascribed to higher brain PCB 77; whereas changes in autonomic functions could be ascribed to higher plasma PCB 180 levels.\u003c/p\u003e \u003cp\u003eConclusion: The pharmacokinetic parameters of PCB 180 and PCB 77 after acute inhalational exposures to Aroclor 1232 were obtained and this was temporally correlated with disruption of behavioral scores and monoamine levels in dose-dependent manner.\u003c/p\u003e","manuscriptTitle":"Behavioral Toxicodynamics and Kinetic profiling of PCB congeners in Swiss albino mouse dosed with Aroclor 1232 through inhalational route: A pivotal risk assessment study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-05 15:58:29","doi":"10.21203/rs.3.rs-4715900/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"3c516fed-0194-4216-b01e-21b24169da3b","owner":[],"postedDate":"August 5th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-06-25T05:38:39+00:00","versionOfRecord":[],"versionCreatedAt":"2024-08-05 15:58:29","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4715900","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4715900","identity":"rs-4715900","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}