%PDF-1.5 % 2 0 obj <>stream BX q 7 0 595 842 re W n 1 0 0 1 7 0 cm endstream endobj 3 0 obj <>stream BX q 0.62293 0 0 0.62293 42.75414 621.16433 cm 2 19.32311 785 218 re W n 1 1 1 rg 2 -17 786 254 re f q 2 218 786 19 re W n BT /EBYNZC 16 Tf 0 0 0 rg 1 0 0 1 34 223 Tm -0.16 Tc 0.16 Tw (CARBONIC ANHYDRASE IN MUSSEL ) Tj ET Q q 2 218 786 19 re W n BT /SXJNNL 16 Tf 0 0 0 rg 1 0 0 1 318 223 Tm 0.14733 Tc -0.14733 Tw (MYTILUS GALLOPROVINCIALIS) Tj ET Q q 2 218 786 19 re W n BT /EBYNZC 16 Tf 0 0 0 rg 1 0 0 1 562 223 Tm 97.04803 Tz 0.11808 Tw (: A PRELIMINARY STUDY) Tj ET Q q 2 183 786 34 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 263 204 Tm 96.67674 Tz 0.56193 Tw (Dijana pavicic Hamer ) Tj ET Q q 2 183 786 34 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 401 204 Tm 97.3443 Tz 0.46612 Tw (* and Bojan Hamer ) Tj ET Q q 2 183 786 34 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 526 204 Tm ( ) Tj ET Q q 2 183 786 34 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 256 187 Tm 96.20967 Tz 0.39214 Tw ( "Ruder Bo) Tj ET Q q 2 183 786 34 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 322 187 Tm 0.165 Tc (\232) Tj ET Q q 2 183 786 34 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 328 187 Tm 93.65832 Tz 0.97563 Tw (kovic" Institute ) Tj ET Q q 2 183 786 34 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 419 187 Tm 0.005 Tc (-) Tj ET Q q 2 183 786 34 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 424 187 Tm ( ) Tj ET Q q 2 183 786 34 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 428 187 Tm 95.44692 Tz (pavicic@cim.irb.hr) Tj ET Q q 2 164 786 19 re W n BT /BDOPZB 16 Tf 0 0 0 rg 1 0 0 1 2 169 Tm (\240) Tj ET Q q 2 45 786 119 re W n BT /EBYNZC 15 Tf 0 0 0 rg 1 0 0 1 2 151 Tm 0.02313 Tc (Abstract) Tj ET Q q 2 45 786 119 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 58 150 Tm ( ) Tj ET Q q 2 45 786 119 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 2 134 Tm 96.62034 Tz 0.60709 Tw (We investigated the esterase activity of carbonic anhydrase \050CA\051 on total esterase activity level in mussels sampled from 19 locations ) Tj ET Q q 2 45 786 119 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 2 117 Tm 96.74238 Tz 1.79817 Tw (along the Croatian coast of Adriatic Sea. The results of total esterase activities in winter were lower than in summer at almost all ) Tj ET Q q 2 45 786 119 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 2 100 Tm 95.84308 Tz 1.58514 Tw (investigated locations. CA activities determined in gills of mussel sampled in winter ranged from 1.75\045 to 24.65\045 of total esterase ) Tj ET Q q 2 45 786 119 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 2 83 Tm 96.67581 Tz 1.71212 Tw (activity. CA activities in summer samples were practically not detectable. Despite ) Tj ET Q q 2 45 786 119 re W n BT /DGLDHA 15 Tf 0 0 0 rg 1 0 0 1 498 83 Tm 0.14214 Tc 1.10786 Tw (in vitro) Tj ET Q q 2 45 786 119 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 544 83 Tm 96.28701 Tz 1.46708 Tw ( research showed potential application of ) Tj ET Q q 2 45 786 119 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 2 66 Tm 95.69378 Tz 1.41148 Tw (CA activity) Tj ET Q q 2 45 786 119 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 70 66 Tm 96.0518 Tz 1.36462 Tw ( in bioassay and biomarker studies for application of CA as a biomarkers of environmental pollution further research are ) Tj ET Q q 2 45 786 119 re W n BT /BDOPZB 15 Tf 0 0 0 rg 1 0 0 1 2 49 Tm -0.03286 Tc (needed.) Tj ET Q q 2 29 786 15 re W n BT /DGLDHA 13 Tf 0 0 0 rg 1 0 0 1 2 33 Tm 0.25639 Tc -0.50639 Tw (Keywords: Adriatic Sea, Ecotoxicology, Enzymes, Mollusca, Pollution) Tj ET Q 0 0 0 RG 2 J [ ] 0 d 2.5 30.5 m 382.5 30.5 l S q 395 204 268 17 re W n BT /BDOPZB 13 Tf 0 0 0 rg 1 0 0 1 395 209 Tm 92.30769 Tz (1) Tj ET Q q 520 204 268 17 re W n BT /BDOPZB 13 Tf 0 0 0 rg 1 0 0 1 520 209 Tm 92.30769 Tz (1) Tj ET Q q 2 187 503 17 re W n BT /BDOPZB 13 Tf 0 0 0 rg 1 0 0 1 250 192 Tm 92.30769 Tz (1) Tj ET Q Q EX endstream endobj 4 0 obj <>stream BX q 0.62293 0 0 0.62293 42.75414 72.16433 cm 2 19.47648 785 868 re W n 1 1 1 rg 2 -80.52352 786 967.52352 re f q 2 -80.52352 388 967.52352 re W n BT /EBYNZC 12 Tf 0 0 0 rg 1 0 0 1 2 877 Tm (INTRODUCTION ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 101 877 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 862 Tm -0.02347 Tc 0.11438 Tw (Carbonic anhydrase \050CA\051, a ubiquitous enzyme in the bacteria, plant, and animal ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 847 Tm -0.00698 Tc 1.00698 Tw (kingdoms catalyses the reversible hydration of CO) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 256 847 Tm 0.1348 Tc 0.8652 Tw ( to produce H) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 332 847 Tm 0.05667 Tc 0.44333 Tw ( and HCO) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 390 847 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 832 Tm 0.00122 Tc 3.79878 Tw (using zinc as a cofactor. To date 15 CAs or CA) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 267 832 Tm 0.004 Tc (-) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 271 832 Tm -0.0488 Tc 3.0488 Tw (like proteins have been ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 817 Tm -0.03098 Tc 0.66735 Tw (identified in mammals. In humans, for CAII isozyme, turnover number for CO) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 390 817 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 802 Tm 0.05905 Tc 1.27429 Tw (hydration is the highest known for any enzyme, while for the other isozymes ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 787 Tm -0.02952 Tc 0.49619 Tw (activities are lower in the order CAII \076 CAIV \076 CAI \076 CAIII. There are several ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 772 Tm 0.0946 Tc 4.57207 Tw (CAs which have shown esteraze activity, enzymes known to hydrolyse ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 757 Tm 0.06525 Tc 1.26808 Tw (endogenous substrates, and the majority hydrolyse lipid ester substrates. In ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 381 757 Tm 96.40103 Tz (in ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 2 742 Tm 0.0664 Tc (vitro) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 25 742 Tm 0.02533 Tc 5.87467 Tw ( conditions, the esterase activity of CA was determined, both in ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 727 Tm -0.03149 Tc 1.19816 Tw (physiological and pathological conditions. Additionally, besides Na,K) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 344 727 Tm 0.004 Tc (-) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 348 727 Tm 0.144 Tc (ATPase, ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 712 Tm 0.01138 Tc 0.65529 Tw (CA represents a key enzyme involved in the adaptation of marine organisms to ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 697 Tm -0.06034 Tc 0.56034 Tw (environmental conditions \1331\135. Experimental studies performed on Crustacea gills ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 682 Tm -0.02133 Tc 3.13244 Tw (have shown two main isozymes of CA located in membrane) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 318 682 Tm 0.004 Tc (-) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 322 682 Tm -0.0664 Tc 3.0664 Tw (bound and in ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 667 Tm -0.05503 Tc 1.55503 Tw (cytosolic fractions, functionally similar to mammalian CAIV and CAII \1332\135. So ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 652 Tm 0.0687 Tc 0.21702 Tw (far no report has dealt with the esterase activity of CA on total esterase activity ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 637 Tm 97.10145 Tz 0.13043 Tw (level in mussel. ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 78 637 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 622 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /EBYNZC 12 Tf 0 0 0 rg 1 0 0 1 2 607 Tm 97.17222 Tz 0.12725 Tw (MATERIAL AND METHODS ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 160 607 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 592 Tm 0.07766 Tc 1.92234 Tw (In this study the esterase activity of CA on total esterase activity level was ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 577 Tm -0.04365 Tc 0.49819 Tw (investigated in mussels sampled from 19 locations along the Croatian coast. The ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 562 Tm 0.03698 Tc 2.96302 Tw (gills were the target tissue because the respiratory, ionic transport and pH ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 547 Tm -0.01733 Tc 2.10067 Tw (regulatory enzyme function of CA and it's potential usage as a biomarker of ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 532 Tm 0.00535 Tc 2.19465 Tw (environmental pollution was the main topic. Previously it was suggested the ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 517 Tm 0.03707 Tc 2.24865 Tw (possible application of CA activity inhibition as an ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 267 517 Tm -0.00057 Tc 2.00057 Tw (in vitro) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 304 517 Tm 0.07257 Tc 1.92743 Tw ( bioassay for the ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 502 Tm -0.03509 Tc 3.47953 Tw (detection of heavy metals in pollution monitoring using the mussel ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 355 502 Tm 97.22222 Tz (Mytilus ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 2 487 Tm -0.05929 Tc (galloprovincialis) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 83 487 Tm -0.01792 Tc 1.35126 Tw ( Lamarck, 1819 \1333,4\135. Total esterase activity was measured in ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 472 Tm -0.03853 Tc 2.03853 Tw (the cytosolic fraction of gills homogenates by colorimetric end point reaction ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 457 Tm -0.00067 Tc 1.00067 Tw (using p) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 38 457 Tm 0.004 Tc (-) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 42 457 Tm 0.07751 Tc -0.07751 Tw (nitrophenyl acetate as enzyme substrate, and CA activity was estimated ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 442 Tm -0.03869 Tc 0.03869 Tw (by the same enzymatic reaction using acetazolamide as a specific CA inhibitor. ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 383 442 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 427 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 234 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 219 Tm -0.02702 Tc 2.8452 Tw (Fig. 1. Total esterase activity in cytosolic fraction of mussel gills: \050) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 353 219 Tm 0.004 Tc (-) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 357 219 Tm 0.585 Tc (AZM\051 ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 204 Tm 0.0311 Tc -0.0311 Tw (without specific CA inhibitor and \050+AZM\051 with acetazoamide inhibition.) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 355 204 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 189 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 174 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /EBYNZC 12 Tf 0 0 0 rg 1 0 0 1 2 159 Tm -0.09771 Tc (RESULTS ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 59 159 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 144 Tm -0.02555 Tc 5.52555 Tw (The results of total esterase activities in winter \050March; average value ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 129 Tm (0.137) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 29 129 Tm 0.232 Tc (\261) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 36 129 Tm -0.02909 Tc 3.47354 Tw (0.057\051 were lower then determined for the summer season \050August; ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 114 Tm (0.153) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 29 114 Tm 0.232 Tc (\261) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 36 114 Tm -0.05159 Tc 5.49603 Tw (0.036\051 at almost all investigated locations \050Fig. 1\051. CA activities ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 99 Tm -0.06719 Tc 0.98385 Tw (determined in gills of mussels sampled in winter ranged from 1.75\045 to 24.65\045 ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 84 Tm 0.02493 Tc 0.79325 Tw (of total esterase activity. CA activities in summer samples were practically not ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 69 Tm -0.02991 Tc 4.21173 Tw (detectable. This is preliminary research; it is difficult to assume a direct ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 54 Tm 0.01619 Tc 5.35881 Tw (relationship between pollution at investigated sites with determined CA ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 39 Tm -0.01651 Tc 0.01651 Tw (activities, especially in the summer period. ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 210 39 Tm ( ) Tj ET Q q 2 -80.52352 388 967.52352 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 24 Tm ( ) Tj ET Q q 380 0 0 172 6 247 cm /LCAGPR Do Q q 251 839 5 15 re W n BT /BDOPZB 10 Tf 0 0 0 rg 1 0 0 1 251 844 Tm (2) Tj ET Q q 326 846 6 15 re W n BT /BDOPZB 10 Tf 0 0 0 rg 1 0 0 1 326 851 Tm 0.36 Tc (+) Tj ET Q q 382 839 5 15 re W n BT /BDOPZB 10 Tf 0 0 0 rg 1 0 0 1 382 844 Tm (3) Tj ET Q q 387 846 3 15 re W n BT /BDOPZB 10 Tf 0 0 0 rg 1 0 0 1 387 851 Tm 90.09009 Tz (-) Tj ET Q q 385 809 5 15 re W n BT /BDOPZB 10 Tf 0 0 0 rg 1 0 0 1 385 814 Tm (2) Tj ET Q Q EX endstream endobj 5 0 obj <>stream BX q 0.62293 0 0 0.62293 299.25414 72.16433 cm 2 147.47648 785 740 re W n 1 1 1 rg 2 130 786 857.47648 re f q 2 157 388 830.47648 re W n BT /EBYNZC 12 Tf 0 0 0 rg 1 0 0 1 2 877 Tm 0.1492 Tc -0.1492 Tw (INSTEAD OF CONCLUSIONS ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 172 877 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 862 Tm 0.19086 Tc (Inspite ) Tj ET Q q 2 157 388 830.47648 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 42 862 Tm -0.00057 Tc 3.00057 Tw (in vitro) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 80 862 Tm 0.00688 Tc 2.88201 Tw ( studies showing clear inhibition of CA activity with heavy ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 847 Tm -0.00921 Tc 1.75921 Tw (metals, for the application of CA as a biomarker of pollution further detailed ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 832 Tm -0.04611 Tc 1.04611 Tw (research on the effects of environmental and physiological factors and different ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 817 Tm 0.02006 Tc 2.60494 Tw (CA isozymes activity using different inhibitors, protein characterization and ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 802 Tm -0.0141 Tc 0.5141 Tw (sequencing are needed. With this paper we report a partial CAII sequence of ) Tj ET Q q 2 157 388 830.47648 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 377 802 Tm 0.002 Tc (M. ) Tj ET Q q 2 157 388 830.47648 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 2 787 Tm -0.05929 Tc (galloprovincialis) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 83 787 Tm -0.03322 Tc 2.48777 Tw ( \050Fig. 2\051 which is, according to the NCBI database the first ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 772 Tm 97.88862 Tz 0.77763 Tw (normal one for Mollusca, Bivalvia, besides nacrein, nacrein) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 288 772 Tm 0.004 Tc (-) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 292 772 Tm 0.01911 Tc -0.01911 Tw (like proteins and the ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 757 Tm -0.06 Tc 0.06 Tw (novel CA from ) Tj ET Q q 2 157 388 830.47648 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 77 757 Tm -0.00062 Tc 0.00062 Tw (Tridacna gigas ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 153 757 Tm 0.07812 Tc -0.07812 Tw (with two CA domains.) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 263 757 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 742 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 727 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 419 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 404 Tm -0.07393 Tc 4.27393 Tw (Fig. 2. Alignment of our partial ) Tj ET Q q 2 157 388 830.47648 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 175 404 Tm -0.05284 Tc 3.05284 Tw (M. galloprovincialis) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 275 404 Tm 0.02275 Tc 2.97725 Tw ( cDNA sequence with ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 389 Tm -0.02659 Tc 0.02659 Tw (similar human and fish CAs after NCBI blast.) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 221 389 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 374 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 359 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 344 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /EBYNZC 15 Tf 0 0 0 rg 1 0 0 1 2 326 Tm 0.504 Tc (References) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 77 327 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 312 Tm (1 ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 12 312 Tm 0.004 Tc (-) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 16 312 Tm 96.24094 Tz 1.11277 Tw ( Pavicic) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 54 312 Tm 0.004 Tc (-) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 58 312 Tm -0.02864 Tc 0.47309 Tw (Hamer D., Devescovi M.\240and Lucu C.,\240 2003. Activation of carbonic ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 297 Tm 0.01686 Tc 0.126 Tw (anydrase in brachial cavity tissues of lobsters \050) Tj ET Q q 2 157 388 830.47648 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 228 297 Tm 0.1672 Tc -0.1672 Tw (H. gammarus) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 295 297 Tm 0.04071 Tc -0.04071 Tw (\051 by dilute seawater ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 282 Tm -0.03556 Tc (exposure. ) Tj ET Q q 2 157 388 830.47648 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 51 282 Tm 96.46302 Tz (JEMBE) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 87 282 Tm -0.056 Tc 0.056 Tw ( 287: 79) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 126 282 Tm 0.004 Tc (-) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 130 282 Tm (92.) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 145 282 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 267 Tm (2 ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 11 267 Tm 0.004 Tc (-) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 15 267 Tm -0.02964 Tc 0.02964 Tw ( Henry R.P., 2001. Environmentally mediated carbonic anhydrase induction in ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 252 Tm -0.07175 Tc 0.07175 Tw (the gills of euryhaline crustaceans. ) Tj ET Q q 2 157 388 830.47648 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 169 252 Tm 96.24094 Tz 0.11277 Tw (J. Exp. Biol.) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 226 252 Tm -0.084 Tc 0.084 Tw ( 204:) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 250 252 Tm ( 991) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 271 252 Tm 0.004 Tc (-) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 275 252 Tm (1002.) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 302 252 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 237 Tm (3 ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 13 237 Tm 0.004 Tc (-) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 17 237 Tm 0.00722 Tc 2.21501 Tw ( Lionetto M.G.,\240 Caricato R.,\240 Erroi E., Giordana M.E.\240 and\240 ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 327 237 Tm 0.02767 Tc 0.97233 Tw (Schettino T., ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 222 Tm 0.01612 Tc 3.65054 Tw (2006. Potential application of carbonic anhydrase activity in bioassay and ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 207 Tm -0.03859 Tc 0.03859 Tw (biomarker studies. ) Tj ET Q q 2 157 388 830.47648 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 93 207 Tm -0.08716 Tc 0.08716 Tw (Chemistry and Ecology) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 204 207 Tm 97.4805 Tz 0.07558 Tw ( 22: S119) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 249 207 Tm 0.004 Tc (-) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 253 207 Tm 97.57155 Tz (S125.) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 280 207 Tm ( ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 192 Tm (4 ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 13 192 Tm 0.004 Tc (-) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 17 192 Tm 0.00063 Tc 1.19937 Tw ( Gilbert A.L. and\240 Guzman H.M., 2001. Bioindication potential of carbonic ) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 177 Tm -0.02584 Tc 2.42584 Tw (anhydrase activity in anemones and corals. ) Tj ET Q q 2 157 388 830.47648 re W n BT /DGLDHA 12 Tf 0 0 0 rg 1 0 0 1 222 177 Tm 96.60362 Tz 1.60189 Tw (Marine Pollution Bulletin) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 345 177 Tm -0.056 Tc 1.056 Tw ( 42: 742) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 386 177 Tm 0.004 Tc (-) Tj ET Q q 2 157 388 830.47648 re W n BT /BDOPZB 12 Tf 0 0 0 rg 1 0 0 1 2 162 Tm (744.) Tj ET Q 0 0 0 RG 2 J [ ] 0 d 386.5 718.5 m 6.5 718.5 l S 0 0 0 RG 2 J [ ] 0 d 5.5 718.5 m 5.5 432.5 l S 0 0 0 RG 2 J [ ] 0 d 386.5 718.5 m 386.5 433.5 l S 0 0 0 RG 2 J [ ] 0 d 386.5 432.5 m 5.5 432.5 l S q 380 0 0 285 6 433 cm /PVQTYX Do Q Q EX endstream endobj 6 0 obj <>stream Q EX endstream endobj 7 0 obj <>stream q BT 0 Tr /Fabc4 8 Tf 0 0 0 rg 1 0 0 1 291.5 43.6 Tm 0 Ts (751) Tj ET Q endstream endobj 8 0 obj <>stream q BT 0 Tr /Fabc1 6 Tf 0 0 0 rg 1 0 0 1 452.338 35.2 Tm 0 Ts (Rapp. Comm. int. Mer M\351dit., 39, 2010) Tj ET Q endstream endobj 9 0 obj <>stream xwVՙ)ɤܹ鹓ɝdL21MFcA "UX@E bX $"5D|9+p?gWyֳ[kwQdʔ)SH:ĭz3'7l`:7dڵQ믿&3zW9rj|MJ<$2mqN)N֯_-S(%M;Ү %KíBSөG " jB[sS\ZU]ӟdSqy=~Ҩi.j6PqE9\^, R?DS$kpJiDWD3a~EL2eʔ q 8Kb_ R(8%%*hʘA$Po[5th)D" 8,n^#lV 24e9Rl Eb)t\nЮfZJV\)4 4mǣ݃O#Nǜ90 ct#cw@L2eʔc^v R]pvMR(%b@ 宝RC`f!aeJu!wVTJR4"64\ybAYʕ|,fk~zf5N99FkD5`[uEjl0deiWJoNV?UIZyz2eʔ)Sq*yH/an͚55\.t%9(dEとIZ$P`E;!lW^Q nh`iv].@fk|ͥX)qQS)'2ݜSTW-蠓?"N$zjjJ ]ۣF$MB T: MσTEկ(>Ώ)WmK* ɔ)SL:ũX6[lvZHDaw29qR]\\,kФr Hoĉ^`%>#wl?U IVZ%(%"޸XU"ɻZ˥/8RF>6l8ٴDf6Gm@h@:8ۊYX7W>CszL8jJj(G<4ؖܮYYbKrΈΨEb XzZXL2eԆT/NLi=В%K{[x1O<yҥ>{wpXz5!~]wq KE'e n~esD￟@C!tJA?G[}D2tYA]flDP}l2zFz\]ݥrzѢE ? 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