Environ. Sci. Technol. l W 4 , 28, 2295-2305
Determination of PCB Metabolic, Excretion, and Accumulation Rates for Use as Indicators of Biological Response and Relative Risk John F. Brown, Jr.' GE Corporate Research and Development, P.O. Box 8, Schenectady, New York 12301-0008
New and old kinetic and chromatographic data were used to calculate, for 146 PCB congeners, the relative rates of metabolism by both P4501A- and P4502B-like cytochromes and the relative human accumulations after 70yr dosing [RHA(70) values] in both normal and chloracnegenic persons. These results permitted establishment of structure-activity relationships, detection of dechlorination as a minor metabolic pathway, characterization of inter-species differences in metabolic response (e.g., P4501A induction), and rationalization of inter-Aroclor differences in chronic toxicity. They also indicated that for environmental PCB mixtures the now readily calculable RHA(70) values should provide an improved measure of relative chronic health risk, and one that should be particularly useful in the evaluation of bioremediation processes. Introduction Commercial polychlorinated biphenyl (PCB) compositions bearing such trademarks as Aroclor, Phenoclor, Clophen, Kanechlor, etc. were once used extensively in nonflammable industrial fluids, resins, and waxes and are now widely distributed in the environment. Today, PCBs are contaminants of concern at about one-third of all Superfund sites as well as at hundreds of local spill or plant sites. Determinations of both the needs for remedial action at such locations and also the relative merits of alternative remediation technologies are generally based upon assessment of presumptive risks to human health. Although a variety of possible human health effects of PCBs have been discussed over the years, site-specific risk assessments in practice generally presume that the end point of concern is cancer, that the human cancer risk may be calculated from the incidence of hepatic tumors in rats dosed with Aroclor 1260, and that all PCB compositions pose cancer risks identical to that of the heavily chlorinated Aroclor 1260. This latter presumption was apparently adopted purely as a matter of administrative convenience (1);it has been long recognized that the less heavily chlorinated and less persistent PCB compositions produce only mild and reversible toxic effects on rat liver tissue (2),whereas the more heavily chlorinated and more persistent compositions produce extensive tissue damage ultimately resulting in liver cancer (3-7). This suggests that PCB cancer risk might be correlated with persistence, i.e., tendency toward chronic accumulation in the body. Before such a concept could be actually used in practical risk assessment, however, it would be necessary to have (a) the body of pharmacokinetic data needed to calculate the relative accumulation tendency for any PCB composition, (b) an evaluation of that data body for chemical self-consistency, and (c) at least preliminary indications that the relative
* E-mail address:
[email protected];c/o Marsha Berry (secretary). 0013-936X/94/0928-2295$04.50/0
0 1994 American Chemical Society
accumulation tendency of PCB compositions did indeed correlate with chronic health risks. Accordingly, the objective of the present study was to develop and evaluate the sets of data on PCB congener clearance rates (which determine accumulation tendencies) that would be needed to implement such an approach. PCB clearance from animals occurs both by nonmetabolic routes (e.g., fecal or respiratory excretion) and by metabolism (e.g., conversion to more readily excreted oxidation products). With gill-breathing animals (e.g., fish), there is often some PCB metabolism (81, but most PCB clearance occurs by simple extraction from the (lipoidal) gill membranes into the surrounding water, so that the relative bioaccumulabilities of most PCB congeners are closely related to their octanol-water distribution coefficients (Kow values) (9). Conversely, with pulmonate (lung-breathing)animals, most PCB clearance occurs by oxidative metabolism (IO),and accumulability is inversely proportional to metabolizability. It is likely that several of the 40-60 different cytochrome P450 (CYP450) isozymes that are believed (11) to be inducible in higher animals participate in PCB metabolism, but the most commonly encountered ones may be operationally divided into two broad classes on the basis of their congener selection patterns. These classes have been designated as either P4501A-like or P4502B-like (8),based on the similarities of their selection patterns to those exhibited in vitro by purified (12) or enriched (13) preparations of either CYP4501A1 (i.e., the isozyme induced by polycyclic aromatic hydrocarbons, some dioxins, PCDFs, and non-ortho-substituted PCBs) or CYP4502B1 (i.e., the isozyme induced by phenobarbital, DDT, and most ortho-substituted PCBs), respectively. Representative examples of the PCB tissue residues remaining after the metabolism of various Aroclors by either P4502B-like or combinations of P4501A- and 2Blike cytochromes are presented in Figure 1. This figure shows that these enzymes can extensively metabolize most of the lower PCBs, many other environmental chlorocarbons, and even some of the higher PCBs. Characteristic features of a P4502B-like clearance pattern are extensive losses of all PCB congeners carrying adjacent meta and para hydrogens and much slower losses of those carrying only meta hydrogens, in both cases independently of the degree of ortho substitution, thereby leaving a residue of congeners with either para (4-1 or di-meta (33-) chlorine substitution on both rings (Figure la,c,d,f). P4501A-like patterns of PCB congener removal differ from the P4502Blike ones in that only non-ortho and mono-ortho chlorinated congeners (designated by asterisks on Figure lb,e) undergo attack and hence appear at sharply reduced levels. The differences between the clearance patterns of Figure l a vs l b or of Figure Id vs l e arise because a number of the higher non-ortho and mono-ortho PCB congeners, which are present in Aroclors 1242 and 1254 but not in 1016, can readily induce CYP4501A isozymes in the rat Environ. Sci. Technol., Vol. 28, No. 13, 1994
2295
8!
a. Aroclor 1016 x Rat P45028
d. Aroclor x Mouse1254 P4502B
b. Aroclor 1242 x Rat P4501A + 2 8
e. Aroclor 1254 x Rat P4501A
i
I
c. Environmental mixture xCrabP45028
,
+ 20
t. Aroclor 1260 x Human P45028
j3 I
Flgure 1. DB-1 gas chromatograms of PCB compositions before (open peaks) and after (solid peaks) metabolism in various species with vertical scales adjusted to permit match of most persistent peak(s) present. Peaks given by P4501A-sensitive mono-ortho-congeners denoted by asterisks. (a) Aroclor 1016 X rat P4502B; rat adlpose PCBs after 3-month dosing at 50 ppm in diet (Mayes). (b) Aroclor 1242 X rat P4501A 4- 28; rat adipose PCBs after 3-month dosing at 100 ppm in diet (Mayes); (c) mixed Aroclors and other chlorocarbons X crab P4502B; blue crab (Callinecfessapidus) from Jamaica Bay, New York, NY, vs local sediment nonpolar chlorocarbons (Shephard). (d) Aroclor 1254 X mouse P45028; mouse adipose PCBs 24 weeks after single dose (redrawn from ref 17). (e) Aroclor 1254 X rat P4501A 2B; rat adipose PCBs after 3-month dosing at 100 ppm in diet (Mayes). (f) Aroclor 1260 X human P4502B; transformer worker serum 25 yr after direct occupational exposure (ref 19).
+
(14), whereas any such induction would appear to be less extensive in the mouse (Figure Id). Our reviews of published and unpublished gas chromatograms of tissue PCB residues indicate that P4501Alike clearance patterns are exhibited by about 40 94 of the teleost fish species examined to date (8) and by some porpoises (15). P4502B-like patterns are exhibited by a few species of fish (B), about half of the crustaceans examined to date (B), and many species of birds and terrestrial mammals, including Aroclor-exposed sheep (16), mice (17), and factory workers (18,19). Combinations of P4501A-like and 2B-like clearance patterns are exhibited by some wildlife species, by most Aroclor-dosed rats (21, and by the human chloracne patients (18, 20-22) who ingested PCB contaminated with the CYPlAl-inducing PCDFs (22). Conversely, published chromatograms and data for seals (23), polar bears (24), and rhesus monkeys (25) show some features that are different from those of the P4501A-likeand P4502B-like patterns as defined above (and also different from each other), suggesting that in these species contributions to PCB metabolism may arise 2296
Environ. Sci. Technol., Vol. 28,No. 13, 1994
from isozymes other than CYP4501Al and CYP4502Bl. In short, PCB clearance patterns vary somewhat with both the species and its toxicological state; in order to properly describe the relative clearance rate of an individual PCB congener, it is necessary to specify which types of metabolic activity are operative. Available data (12, 13, 18, 19, 21, 22) on the relative clearance rates of individual PCB congeners in systems showing either P4501A- and/or P4502B-like selection patterns are generally restricted to those congeners that (a) gave the stronger peaks on a gas chromatogram and (b) fell in the particular range of clearance rates that produced readily quantifiable changes under the conditions of observation. Accordingly, in order to develop sets of clearance rate constants for all of the congeners resolved by current high-resolution capillary GC procedures, it was necessary first to review the original chromatograms, to permit estimates of the clearance of minor as well as major peaks, and second to overlap the data sets from systems covering different ranges of PCB congener reactivities in order to create single sets of relative metabolizabilities
Table 1. Characteristics of Kinetic and Chromatographic Data Sets Used for Calculating Relative Rates of PCB Congener Metabolism relative P450 activity abbr
metabolic system
pattern type
RlAl R2B1 RMMC RMPB ATC AME MUS HYPl HYP2 HCW
rat liver P4501A1, purified (12) rat liver P4502B1, purified (12) rat liver microsomes, 3-MC-induced (13) rat liver microsomes, PB-induced (13) Atlantic tomcod, 3.6 ppm PCB (8) American eel, 100 ppm PCB (8) mouse, dosed with Aroclor 1254 (17) yucheng patients exposed to PCB/PCDF (22) yucheng patients exposed to PCB/PCDF (23) capacitor workers, exposed to Aroclors 1016, 1242, and 1254 (18) transformer worker, exposed to Aroclor 1260 (19)
HTW
kn (yr-9
Sa
sb
1A 2B 1A 2B 1A 2B 2B 1A + 2B 1A 2B 2B
0 0 0 0 (for Cl4 BP) (for C15 BP)
3.5 X lo4 0 53 0 5.3 x 1 0 4 b -0 -0
0 1.3 x 105 0 2.6 x 103
0.14 0.14 0.008
0.7 0
1.0 1.0 1.0
2B
0.008
0
1.0
+
data type
1.0
o
7.8 X lo5 b
15
Ratio of apparent first-orderclearance rate of an individual PCB congener in system S to that of same congener in reference system. * Value required to convert human (ski - kn) in yr-l to fish ki/kn.
covering all PCB congeners for each type of metabolic activity. Materials and Methods
Calculation of Kinetic Parameters. We shall assume that the uptake, clearance, and accumulation of PCBs in an animal may be described by eqs 1-6:
dAi/dt = kdi - kiAi
(mass balance)
Ai@) = (kdi/k,) [l - exp(-kit)l
+ A,(O) exp(-kit)
(3)
(integration of eq 3) (4) of eq 4 terms) (5) APCB(t)+ k ~ ~ ( t (summation ) RHA(t) = ApcB(t)/Al,,,(t)
(definition)
(6)
where ki is the observed clearance rate for individual congener PCBi in system S (yr-l); kai is the relative rate constant for P4501A-type metabolism of PCBi; kbi is the relative rate constant of P4502B-typemetabolism of PCBi; kdi or k p c is ~ the dose rate for PCBi or total PCB (g yr-l); 12, is the nonmetabolic PCB clearance rate (yr-l); sa is the relative activity pf P4501A in system S; Sb is the relative activity of P4502B in system S;ci is the weight fraction of PCBi in PCB dosed; Ai or APCBis the total accumulation of PCBi or total PCB (g); t is the time (yr);RHA(t) is the relative human PCB accumulation at time t , e.g., RHA(70) is the relative human accumulation over a 70-yr lifetime. The first four columns of Table 1identify the 11kinetic or chromatographic studies that were drawn upon as sources of relative rate data along with the metabolic clearance pattern type@) displayed by each. The next column indicates the type of relative rate indices provided by each raw data set: for the kinetic studies, the reported ki (or t1p)values were presumed to include either sakai or Sbkbi terms depending upon whether the congener selection pattern was of type 1A or 2B, respectively, and also a k, term for the in vivo studies. The chromatographic data used were from systems where the original congener distribution was known from either the Aroclor composi-
tion or that of the local environmental background, so that changes in relative peak heights could be determined. The reference peaks used for pulmonate subjects undergoing simple clearance (kdi = 0) were the most slowly metabolized of the readily measured peaks, since the parameter calculable from the observed changes in selected peak heights by eq 4 is the difference in metabolic rate constants. The reference peaks used for the fish were those of nonmetabolizable isomers having about the same KO,value and, hence, the same k, as the measured peak. If the fish were presumed to be at equilibrium (dAi/dt = 0), the ratio of the original peak-to-reference ratio to the observed peak-to-reference ratio minus 1can be shown to be equal to the ratio of the metabolic to nonmetabolic rate constants. The metabolic rates for several moderately slowly cleared PCB congeners in the yucheng patient group HYP-1 that were not listed in the original paper were estimated from the published chromatogram (21). All data on metabolic rates of the more slowly cleared congeners came from our own studies of PCB-exposed capacitor (18, 19, 26, 27) and transformer (19) workers. The ki values for the capacitor workers represent geometric mean values for the first 6 years’ clearance in a group of 38 workers, who had previously had 17 yr (average) exposure to Aroclors 1254, 1242, and 1016 and who had apparently reached steady-state levels before the clearance period that followed discontinuance of PCB use (26). The generally modest individual variations from such population mean values as functions of age, sex, smoking, body fat, and PCB body burden will be described elsewhere. Several of the long-service transformer workers who had had early exposure to Aroclor 1260 exhibited chromatograms indicating extensive 1260 metabolism. For determinations of sbkbi values, however, we elected to use just the one (Figure If) from an individual with a serum PCB level of 334 ppb who had had a direct exposure to Aroclor 1260 about 25 yr previously and no significant PCB exposure subsequently, so that simple clearance (eq 4, k d i = 0) could be assumed. The composition of the PCB mixture found in the feces of chronically Aroclor 1254-dosedmonkeys is substantially identical to that of the blood (25), indicating that the k, values for nonmetabolic elimination via fecal fat must be substantially identical for all PCB congeners and presumably likewise for other highly lipophilic xenobiotics as well. Accordingly, this k n for normal humans, i.e., those not Envlron. Scl. Technol., Vol. 28, No. 13, 1994
2297
Table 2. Congeneric Composition, Metabolic Rates, and Relative Accumulabilities of Analytically Resolvable PCB Components
SE-30 DB-1 peak peak and RRT Clnos. 1-0 11 2-1 3-1 4-1 16 5-2 6-2 7-2 21 8-2 9-2 10-3 11-3 12-2 13-2 14-2 28 14-3 28 15-3 16-3 32 17-3 18-3 19-3 19-4 20-3 21-3 22-3 23-3 37 37 24-3 24-4 25-3 40a 25-4 26-3 40b 26-4 27-4 28-3 29-4 30-3 31-4 47 32-4 47 33-4 47 34-4 47 35-4 36-3 37-4 54 37-5 38-3 38-4 54 39-4 58 40141-4 40141-5 42-4 43-4 43-5 44-4 44-5 45-4 46-4 70 46-5 70 47-4 70 70 48-4 70 48-5 49-4 49-5 78 50-4 50-4 78 51-5 84 52-5 53-5 84 54-5 84 55-5 55-6 56-5 96 57-5 2298
PCB congeners present” 000 (biphenyl) 001 002 003 @,010 007,009 006 005 008 014’019 030 011 012,013 -;015 018;017 0.24,m 016;032 023 034;-;054 029 026 025 031 028 050 020,021,033 053 022 051 045 036 046 039 052,073 049,043
major chlorination pattern
2342-2 24-, 252-3 2-4 3526-2 2463-3 3-4 4-4 25-2 24-2 26-3 23-2,26-4 23535-2 26-26 24525-3 24-3 25-4 24-4 246-2 34-2 25-26 23-4 24-26 236-2 35-3 23-26 35-4 25-25 24-25 24-24 047 048,075 245-2 065,062 235634-3 035 -;044 23-25 104;246-26 34-4 037;-;%,059 23-24 041,064,071,072 236-4,26-34 068 24-35 096 236-26 040 23-23 057 235-3 103 246-25 067 245-3 100 246-24 063,058 235-4 -; 0 2 , 0 6 1 245-4 094;235-26 070 25-34 066,076;24-34 098’ 095 093,102 236-25 234-3 055’-’ 091 236-24 056 23-34 060 234-4 084 092 236-23,235-25 234-26 089’101,090 245-25 099 245-24 119 246-34 150 236-246 083,112; 109 235-23 097,086 245-23
Environ. Sci. Technol., Vol. 28, No. 13, 1994
coeluting congener ratios
0.3b 0.7
0.5b
0.5
systems providing data used (see Table 1)
ka
SAR -3000 SAR -100 SAR -1400 SAR -1400 RlA1, RMMC, R2B1, RMPB -2 RlA1, RMMC, ATC, R2B2, RMPB 100 RMMC, ATC, RMPB 40 RlA1, RMMC, ATC, R2B2, RMPB 60 RlA1, R2B1 500 RMMC, ATC, RMPB 0 SAR 0 RlAI, R2B1 1100 RlA1, (ATC), R2B1 240 RlA1, (ATC), R2B1 250 (RMPB), SAR 0 RMPB, SAR 0 RMPB, SAR 0 RMPB, SAR 0 -50 SAR SAR -50 SAR 0 -60 SAR RMMC, RMPB 40 40 (RMMC), RMPB (RMMC), SAR -50 RMMC, ATC, HCW 10 SAR 10 60 RMPB, SAR 0 ATC,SAR 190 RMMC, ATC, RMPB 0 ATC RMPB 0 -100 SAR 0 RMMC, (ATC), RMPB, (AME) -100 SAR 0.0 RMPB, AME 0.0 RMPB, AME 0.0 HCW 0.0 (RMPB), SAR 0.0 SAR 100 (ATC), SAR 0.0 (RMMC), ATC, RMPB, AME SAR 0 100 (ATC), SAR 0.0 RMPB,AME 0.0 RMPB -3 SAR 0 SAR 0.0 RMPB 50 (HCW), SAR 0.0 (HCW), SAR 30 (HCW), SAR 0.0 (HCW), SAR 3 HYP1, HCW 0.5 HYP1, 2; HCW 0.0 SAR 50 RMMC, ATC, RMPB, AME 3 ATC. HYP1, HCW 0.0 AME 50 SAR 0.0 RMPB,AME 50 (RMPB), (AME), SAR -2 (RMMC), HYP1, HCW 0.0 RMPB, AME, HYPl 0.0 SAR 0.0 RMPB, AME, HYPl 0.0 MUS. HYP1. HCW 0.0 SAR 0.0 SAR 0.0 (HCW), SAR 0.0 RMPB, AME
-
0.9gb 0.01 0.95b 0.05 0.97b 0.03
LOOb
0.00 0.6* 0.4
0.5* 0.5 0.gb 0.1
1.00b 0.00 0.8b 0.2 O.lb 0.9 0.7b 0.3
LOc 0.0 l.Oc
relative accumulative RHA(70) chloracne normal patient human kb -200 0.000 0.000 -200 0.001 0.000 -60 0.000 0.001 -80 0.000 0.001 200 0.001 0.000 60 0.002 0.001 60 0.003 0.001 80 0.002 0.001 6 0.001 0.01 100 0.003 0.001 -80 0.003 0.001 10 0.000 0.006 10 0.001 0.006 0.5 0.001 0.12 -20 0.014 0.003 40 0.007 0.001 34 0.008 0.002 12 0.023 0.005 -6 0.006 0.01 -6 0.006 0.01 -60 0.005 0.001 -2 0.005 0.03 3.6 0.007 0.016 2.4 0.007 0.024 -3 0.006 0.019 0.48 0.028 0.12 -40 0.006 0.001 5.2 0.005 0.011 -20 0.014 0.003 1.8 0.001 0.03 40 0.007 0.001 36 0.008 0.002 -2 0.003 0.03 -20 0.014 0.003 -0.8 0.003 0.07 6.5 0.042 0.009 5.8 0.048 0,010 2 0.14 0.03 2 0.14 0.03 6 0.05 0.01 6 0.003 0.01 6.4 0.042 0.009 0.007 0.001 40 0.8 0.003 0.07 3.4 0.08 0.017 7 0.04 0.008 1 0.07 0.06 40 0.007 0.001 0.041 0.009 6.6 2 0.006 0.03 2 0.14 0.03 2 0.009 0.03 1 0.24 0.06 0.6 0.07 0.1 0.27 0.21 0.35 0.05 0.01 6 0.006 0.05 1.2 0.07 0.07 0.8 0.05 0.01 6 0.006 0.01 6 0.05 0.01 6 0.006 0.03 -2 -1 0.14 0.06 0.17 0.04 1.5 0.05 0.01 6 0.13 0.03 2 1.2 0.28 0.20 0.05 0.01 6 0.05 0.01 6 0.3 0.06 1 0.3 0.06 1
PCB-exposed human
-
-
-
-
Table 2 (Continued)
SE-30 peak RRT 96
104 117
125 125
146 146 146 160
174 174
203 203 203 232 232 244 244
280
332 372 372 372 448 528
DB-1 peak and C1 nos. 57-6 58-5 59-5 60-6 61-4 61-5 62-6 63-5 64-6 65-5 65-6 66-6 67-5 67-6 68-5 69-5 69-6 70-6 71-5 71-6 72-5 72-6 72-6 73-6 74-5 74-6 75-6 76-6 77-6 78-7 79-6 80-6 81-7 82-6 82-6 83-6 84-5 84-6 85-7 86-6 87-7 88-7 89-6 90-7 94-6 92-7 93-7 94-7 95-6 95-7 96-8 97-6 98-7 99-8 100-7 100-8 101-8 102-7 103-7 104-7 105-8 106-7 107-7 108-8 109-8 110-8 110-8 111-7 112-8 113-9 114-9 115-8
PCB congeners presenta 152 087,111; 115 085,116, DDE 136 077; -; 110 154 082 151 124 135 144 107,108 147 123 118,106 149,139 140 114 134,143 122 131,142 133 146 105 132,161 153 168 141 179 137 130 176 138,164,160 163 158,160 126 129 178 166 175 187,182 128 183 167 185 174,181 177 156 171 202 157 173 200 172 204 197 180 193 191 199 170 190 198 201 196 203 189 195 208 207 194 _.
major chlorination pattern 2356-26 234-25 234-24 236-236 34-34 236-34 245-246 234-23 2356-25 345-25 235-236 2346-25 235-34 2356-24 345-24 245-34 236-245 234-246 2345-4 2356-23 345-23 2346-23 235-235 235-245 234-34 234-236 245-245 246-345 2345-25 2356-236 2345-24 234-235 2346-236 234-245 2356-34 2346-34 345-34 2345-23 2356-235 23456-4 2346-235 2356-245 234-234 2346-245 245-345 23456-25 2345-236 2356-234 2345-34 2346-234 2356-2356 234-345 23456-23 2346-2356 2345-235 23456-246 2346-2346 2345-245 2356-345 2346-345 23456-236 2345-234 23456-34 23456-235 2345-2356 2345-2346 23456-245 2345-345 23456-234 23456-2356 23456-2346 2345-2345
coeluting congener ratios 0.0
0.1c 0.9
0.3c 0.7 0.5c 0.5 0.8c 0.2 0.7c 0.3 0.7c 0.2 0.1 0.7c 0.3
0.7d 0.3 O.OOd
1.00
0.3d 0.7
1.00d 0.00
0.5d 0.5
PCB-exposed human
systems providing data used (see Table 1)
kb
ka
SAR RMPB, AME SAR RMPB, AME (ATC), (HCW) RMPB, AME SAR RMPB, AME RMPB, AME, HYPl (RMPB), (AME), SAR (RMPB), (AME), SAR (AME), SAR HYPl SAR SAR ATC, HYP1,2, MUS, HCW (RMPB), AME, SAR SAR HYP1. HCW RMPB SAR SAR (HTW), SAR HTW ATC, HYP1, MUS, HCW AME, SAR MUS, HTW, HYPS, HCW HTW (RMPB), AME, HYPl (RMPB), (AME), SAR HTW SAR (RMPB), (AME), SAR HYP1, HCW, MUS HTW (HTW), SAR HYP1, SAR (RMPB), SAR HTW SAR SAR HTW (MUS), SAR HTW HTW RMPB, AME RMPB, AME HTW (MUS), (HYP1,2), HTW HTW HTW SAR SAR SAR HTW SAR SAR (MUS), (HYPZ), HTW HTW (HTW), SAR RMPB (MUS), (HYPB), HTW HTW HTW HTW IlTW HTW HTW HTW HTW (HTW), SAR HTW
0.0 0.0
0.0 0.0 50 0.0 0.0 0.0 0.0 20 0.0 0.0 -2 0.0 -1 0.60 0.0
-
-
-
0.0 0.15 0.0 20 0.0 0.0 0.0 0.93 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
0.0 0.0 -1 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
0.0 0.0 0.0
0.0 0.0 0.0 0.0 0.0
0.0 0.0 0.0 0.0 0.0
0.0 0.0 0.0
0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
2 3.6 0.8 7.0 >0.2 4.2 0.2 3.4 1.8 2 3 2 0.14 0.8 0.07 0.11 1.4 -0.8 0.04 2 2 2 0.0 0.026 0.17 2 0.046 0.070 1 2 0.024 0.1 2 0.08 -0.03 -0.14 -0.1 1
-
