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WittigMass SpectrometryTop-down proteomicsS-nitrosylationRedox-proteomicshttps://www.ebi.ac.uk/pride/archive/projects/PXD075167LungTwo lungs were excised from each mouse in DMEM supplemented with 10% FCS. The lungs were minced into small pieces and digested in Collagenase A for 30 minutes at 37 °C in a water bath. After digestion, cells were separated from the remaining tissue using a 70 µm cell strainer and washed with MACS buffer (1× PBS, 5% BSA, 2 mM EDTA). The intact red blood cell (RBC) pellet was collected and subjected to density gradient centrifugation using Histopaque 1119 to remove other blood cells from erythrocytes. Endothelial cells were isolated from total pulmonary cells using CD31 MicroBeads and MACS columns. The isolated endothelial cells (ECs) were washed in MACS buffer (1× PBS) containing 100 mM NEM and 0.1 mM neocuproine. Twenty microliters of pEPC were resuspended in 150 µl NO–50 mM NEM-DAB denaturing buffer (8 M urea, 5 mM EDTA, 0.5% SDS, 100 mM Tris/HCl pH 8.5, 50 mM NEM, 100 µM neocuproine, protease inhibitor cocktail). After incubation for 1 h at 22 °C and 850 rpm, insoluble material was removed by centrifugation. Proteins were precipitated with ice-cold acetone, collected by centrifugation, washed, and resuspended in 200 µl ascorbate-DAB buffer (8 M urea, 5 mM EDTA, 0.5% SDS, 50 mM Tris/HCl pH 8.5, 1 mM ascorbic acid, 10 µM CuSO₄). Samples were incubated at 850 rpm for 5 min at 22 °C in the dark, followed by the addition of 200 µl BIAM-DAB buffer (8 M urea, 5 mM EDTA, 0.5% SDS, 50 mM Tris/HCl pH 8.5, 20 mM BIAM [EZ-Link™ Iodoacetyl-PEG2-Biotin, Thermo Scientific]) and incubation for 1 h at 22 °C in the dark at 850 rpm. Proteins were precipitated overnight at −20 °C with ice-cold acetone, collected by centrifugation, washed, and resuspended in 200 µl lysis buffer (5 mM EDTA, 50 mM Tris/HCl pH 8.5, 1% Triton X-100, 1% SDS). A total of 200 µg protein was subjected to affinity purification using agarose streptavidin beads overnight at 4 °C on a rotating wheel. The supernatant from the affinity purification was digested separately for further analysis of overoxidized proteins. After washing, beads were resuspended in 50 µl 6 M guanidine hydrochloride (GdmCl), 50 mM Tris/HCl pH 8.5, and incubated at 95 °C for 5 min. Samples were diluted with 25 mM Tris/HCl pH 8.5 and 10% acetonitrile to obtain a final GdmCl concentration of 0.6 M. Proteins were digested with 1 µg sequencing-grade trypsin (Promega) overnight at 37 °C under gentle agitation. Digestion was stopped by adding trifluoroacetic acid to a final concentration of 0.5%. Peptides were loaded onto multi-stop-and-go extraction tips (StageTips) containing three C18 discs and three SCX discs. Peptide purification and elution were performed as described by Kulak and Mann (2014). Peptides were eluted into microtiter plates, dried, and reconstituted in 1% acetonitrile and 0.1% formic acid. Liquid chromatography–mass spectrometry (LC–MS) was performed using a Thermo Scientific™ Q Exactive Plus mass spectrometer equipped with an ultra-high-performance liquid chromatography unit (Thermo Scientific Dionex Ultimate 3000) and a Nanospray Flex ion source (Thermo Scientific). Peptides were first loaded onto a C18 reversed-phase precolumn (Thermo Scientific) and then separated on an in-house packed picotip emitter (100 µm diameter, 15 cm length; New Objectives) containing 2.4 µm Reprosil C18 resin (Dr. Maisch GmbH). Peptides were eluted using a gradient from mobile phase A (4% acetonitrile, 0.1% formic acid) to 25% mobile phase B (99% acetonitrile, 0.1% formic acid) over 30 min, followed by an increase to 50% B over 5 min. A flow rate of 300 nl/min was applied. MS data were acquired using a data-dependent acquisition Top20 method in positive ion mode, selecting the most abundant precursor ions for higher-energy collisional dissociation (HCD) fragmentation. Full MS scans were recorded over a range of 300–2000 m/z at a resolution of 70,000, with an automatic gain control (AGC) target of 3 × 10⁶ and a maximum injection time of 65 ms. Only multiply charged ions (≥2+) were selected for MS/MS analysis, which was performed at a resolution of 17,500 with an isolation window of 2 m/z, an AGC target of 1 × 10⁵, and a maximum injection time of 150 ms. Previously fragmented ions were excluded for 30 s. Full-scan spectra were acquired in profile mode, and fragment spectra were recorded in centroid mode using Xcalibur software.PrideNot available6-dioxaoctanediamine derivatized cysteineMS Data were analysed by MaxQuant (v2.0.3.0) [Cox and Mann 2008] using default settings. Proteins were identified using reviewed mouse reference proteome database UniProtKB with 71785 entries, released in 8/2023. N-terminal acetylation (+42.01) and oxidation of methionine (+15.99), N-ethylameimide on cysteines (+125.05) and biotinylated iodoacetamide (+414.19) were selected as variable modifications. Lable free quantification (LFQ) and intensity-based absolute quantification (IBAQ) values were recorded. Data were further analysed by Perseus (v. 1.6.1.3). Contaminants and reversed identification were removed. Protein identification with at least 3 valid quantification values in at least one group were further analysed. Missing values were replaced by background values from normal distribution. Student´s t-test and permutation based FDR (q value) was used to identify significant enriched proteins between experimental groups. Rappsilber J, Mann M, Ishihama Y. Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips. Nat Protoc 2, 1896-1906 (2007). Cox J, Mann M. (2008) MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat Biotechnol. 26:1367-72. Tyanova S, Temu T, Sinitcyn P, Carlson A, Hein MY, Geiger T, Mann M, Cox J. (2016)The Perseus computational platform for comprehensive analysis of (prote)omics data. Nat Methods. 13:731-40.ProteomicsProf. Dr. med. Katrin SchäferDepartment of Cardiology, University Medical Center Mainz, GermanyCOMPLETEMus Musculus (mouse)wittig@med.uni-frankfurt.de42085863 Gogiraju R, Sun B, Bochenek ML, Wittig I, Rezende F, Lopez M, Wirth A, Zhao W, Böhm EW, Guliani P, Moiko K, Molitor M, Wenzel P, Freichel M, Lurz P, Brandes RP, Schäfer K. Loss of erythrocyte arginase-1 impairs vasorelaxation due to endothelial GSNOR overexpression and denitrosylation of G protein subunits. Redox Biol. 2026 94:104201 10.1016/j.redox.2026.104201Functional Proteomics, Goethe University, Frankfurt am Main , GermanyGermany10.6019/PXD075167<h4>Introduction</h4>Overexpression of arginase-1 (ARG1) in red blood cells (RBCs) is associated with endothelial dysfunction, and short-term ARG1 inhibition or l-arginine supplementation restored endothelial function. However, the long-term consequences of ARG1 loss in RBCs and its effects on endothelial cells (ECs) are largely unknown. Here, we determined how deletion of ARG1 in RBCs affects blood pressure and vasorelaxation and the role of endothelial NO signaling via the soluble guanylyl cyclase and the S nitrosylation pathway for vascular homeostasis.<h4>Methods</h4>Vascular function was monitored and vasorelaxation studied in C57BL/6 wild-type (WT) and apolipoprotein E deficient (apoE<sup>-/-</sup>) mice lacking ARG1 in cells of the erythrocyte lineage (RBC.ARG1-knockout, KO) and in mice with inducible deletion of ARG1 in ECs (END.ARG1-KO). Primary cells were analyzed using redox proteomics, immunolabeling and fluorescence microscopy.<h4>Results</h4>Loss of ARG1 in RBCs resulted in significantly elevated plasma nitrite and lower mean and diastolic blood pressure levels, and increased phosphorylation of Vasodilator-Stimulated Phosphoprotein in smooth muscle cells indicated overactivated cyclic GMP signaling. In ECs, nitrosoglutathione reductase (GSNOR) overexpression and denitrosylation of endothelial Guanine Nucleotide Binding Proteins was observed, which may have uncoupled NO signaling from cGMP-mediated vasorelaxation in response to acetylcholine. Importantly, inhibition of GSNOR restored the impaired endothelium-dependent vasorelaxation in RBC.ARG1-KO aortas to levels comparable to RBC.ARG1-WT controls.<h4>Conclusions</h4>Our data support the contribution of RBC-derived NO to blood pressure regulation, but also show that chronically elevated circulating NO levels induce counterregulatory mechanisms in ECs, including increased GSNOR expression and protein S-denitrosylation leading to impaired vasorelaxation despite overactivated cGMP signaling.Loss of erythrocyte arginase-1 impairs vasorelaxation due to endothelial GSNOR overexpression and denitrosylation of G protein subunits.Gogiraju Rajinikanth R, Sun Beichen B, Bochenek Magdalena L ML, Wittig Ilka I, Rezende Flávia F, Lopez Melina M, Wirth Angela A, Zhao Wenjia W, Böhm Elsa W EW, Guliani Payal P, Moiko Kateryna K, Molitor Michael M, Wenzel Philip P, Freichel Marc M, Lurz Philipp P, Brandes Ralf P RP, Schäfer Katrin KfalseProtein S-nitrosylation in lung endothelial cells from mouseLong.term consequences of loss of arginase-1 (Arg1) in red blood cells were analysed by redox-proteomics. We enriched S-Nitrosylated proteins to study nitrosylated proteins in red-bood cells using a BIAM-switch assay.2026-06-082026-03-03PXD075167NEWT:3555NEWT:71647NEWT:330879NEWT:260201NEWT:35554NCBITaxon:12939NEWT:32046NEWT:544496NEWT:59529NEWT:2042546NEWT:32049NEWT:1392696NEWT:259447NEWT:295546NCBITaxon:2719036NEWT:45351NEWT:445974NEWT:43179NEWT:180454NEWT:5722NCBITaxon:1280NEWT:112503NEWT:1129NEWT:309807NEWT:10245NEWT:55153NEWT:89184NEWT:309800NEWT:281395NEWT:1211601NEWT:876138NEWT:44271NEWT:193516NEWT:351581NEWT:43186NEWT:428146NEWT:111205NEWT:1117NEWT:498257NEWT:10036NEWT:1590NEWT:498019NEWT:1351NEWT:1438992NEWT:1352NEWT:2649997NEWT:1147161NEWT:661410NEWT:638632NEWT:43150NEWT:263737NEWT:224326NEWT:1333499NEWT:376619NCBITaxon:79857NEWT:1096976NEWT:5702NEWT:95648NEWT:1589NEWT:135622NEWT:1349NEWT:35786NEWT:1580NEWT:399784NEWT:96731NEWT:383379NEWT:1401257NEWT:418106NEWT:10029NEWT:913645NEWT:44491NEWT:641809NEWT:27933NEWT:556484NEWT:317447NEWT:4688NEWT:7955NEWT:111225NEWT:7719NEWT:7959NEWT:2261NEWT:868565NEWT:135674NEWT:94323NEWT:31156NEWT:3112NEWT:79329NEWT:4442NEWT:192875NEWT:1214915NEWT:30069NEWT:12637NEWT:59729NEWT:2164133NEWT:284218NEWT:295105NEWT:108061NEWT:60711NEWT:1316931NEWT:224308NEWT:3347NEWT:511145NEWT:160621NEWT:212790NEWT:931281NEWT:4432NEWT:5762NEWT:5763NEWT:658457NEWT:1310161NEWT:77133NEWT:295358NEWT:145481NEWT:1310165NEWT:29058NCBITaxon:79824NEWT:2246NEWT:2652724NEWT:5757NEWT:68415NCBITaxon:4563NEWT:5755NEWT:44688NEWT:44689NEWT:44447NEWT:752555NEWT:498211NEWT:5759NEWT:347256NEWT:1736231NEWT:5518NEWT:398007NEWT:1527468NEWT:1392NEWT:498217NEWT:498216NEWT:2242NEWT:11320NEWT:286NEWT:391619NEWT:246196NEWT:287NEWT:246197NEWT:145458NEWT:633149NEWT:1149786NEWT:10239NEWT:68895NEWT:44685NEWT:161934NEWT:4897NEWT:1148NEWT:5744NEWT:72901NEWT:213863NEWT:5508NEWT:3329NEWT:5507NEWT:410661NEWT:55571NEWT:35500NEWT:1235816NEWT:1140NCBITaxon:2157NEWT:1143NEWT:4896NEWT:1287689NEWT:1390NEWT:11557NEWT:1094343NEWT:1462472NEWT:1336795NEWT:644042NEWT:294NEWT:1773NEWT:1182590NEWT:3712NEWT:82380NEWT:3711NEWT:935293NEWT:375146NEWT:270643NEWT:2065263NEWT:177437NEWT:1772NEWT:10418NEWT:118698NEWT:1616117NEWT:263NEWT:118696NEWT:34865NEWT:52283NEWT:11988NEWT:284812NEWT:8175NEWT:400667NEWT:43330NEWT:1603293NEWT:408169NEWT:44664NEWT:47946NEWT:459758NEWT:3702NEWT:1245466NEWT:243277NEWT:1246791NEWT:7067NEWT:118694NEWT:586722NEWT:2850NEWT:118691NEWT:34871NEWT:33548NEWT:274NEWT:7070NCBITaxon:287NEWT:408172NEWT:408170NEWT:96794NEWT:3708NEWT:332648NEWT:44670NEWT:536231NEWT:376219NEWT:219813NEWT:1510NEWT:1436733NEWT:460519NEWT:1247411NEWT:1515NEWT:572307NEWT:1432138NEWT:9238NEWT:1424507NEWT:1194599NEWT:272844NEWT:1303443NEWT:9483NEWT:333760NEWT:485NEWT:34607NEWT:56636NEWT:1513458NEWT:1233681NCBITaxon:40559NEWT:506599NEWT:2853422NEWT:81077NEWT:84588NEWT:1679718NEWT:480NEWT:65349NEWT:67767NEWT:46835NEWT:109757NEWT:582580NEWT:300852NEWT:294607NEWT:1502NEWT:128017NEWT:376686NEWT:95486NEWT:508771NEWT:1883446NEWT:253NCBITaxon:2759NEWT:1233435NEWT:93061NEWT:93062NEWT:34613NCBITaxon:4932NEWT:109760NEWT:29031NEWT:943274NEWT:644223NEWT:1263082NEWT:235443NEWT:108458NEWT:5936NEWT:272623NEWT:272624NEWT:320637NEWT:886559NEWT:3750NEWT:55521NEWT:983964NEWT:118499NEWT:11706NEWT:32644NEWT:527796NEWT:1130798NEWT:55529NEWT:499175NEWT:109779NEWT:3745NEWT:1715989NCBITaxon:4751NEWT:476272NEWT:3747NEWT:195051NEWT:3988NEWT:1116234NEWT:561007NEWT:1255228NEWT:649908NEWT:410289NEWT:373153NEWT:3983NEWT:203696NEWT:472NEWT:352472NEWT:97441NEWT:1154756NEWT:1071661NEWT:1202531NEWT:1202532NEWT:360094NEWT:1408223NEWT:470NEWT:1408224NEWT:41364NEWT:5911NEWT:1313NEWT:1218097NEWT:1080348NCBITaxon:50557NEWT:1314NEWT:39251NEWT:39491NEWT:212142NCBITaxon:5811NEWT:29491NEWT:101841NEWT:153481NEWT:2014887NEWT:1499105NEWT:33952NEWT:445NEWT:153009NEWT:261756NEWT:571256NEWT:40483NEWT:63366NEWT:63367NEWT:688333NEWT:215402NCBITaxon:548681NEWT:318586NCBITaxon:5820NEWT:1547NEWT:272634NEWT:198107NEWT:9031NEWT:27292NEWT:1872122NEWT:1308NEWT:7091NEWT:1034015NEWT:760568NEWT:141262NEWT:108931NEWT:1293497NEWT:1055524NEWT:4087NEWT:9778NEWT:306NEWT:150475NEWT:303NEWT:267872NEWT:97477NEWT:7111NEWT:347515NEWT:172269NEWT:9534NEWT:6269NEWT:5180NEWT:256737NEWT:4090NEWT:4092NEWT:9541NEWT:8694NEWT:4093NEWT:33936NEWT:4096NEWT:8692NEWT:2903NEWT:185579NEWT:941442NEWT:536088NEWT:13076NEWT:1226408NEWT:7108NEWT:40479NEWT:43989NEWT:4076NEWT:142615NEWT:317NEWT:1006581NEWT:33940NEWT:885318NEWT:39488NEWT:550NEWT:4081NEWT:273068NEWT:554NEWT:98334NEWT:451516NEWT:552NEWT:231663NEWT:4084NEWT:1325291NEWT:63577NEWT:36185NEWT:1225786NEWT:7574NEWT:1715256NEWT:30640NEWT:575412NEWT:929793NEWT:1507179NEWT:29204NEWT:28112NEWT:114155NEWT:6493NEWT:6494NEWT:6491NEWT:507601NEWT:520NEWT:186441NEWT:634955NEWT:643680NEWT:214092NCBITaxon:6157NEWT:6239NEWT:13095NEWT:162425NEWT:104105NEWT:216257NEWT:102169NEWT:208893NEWT:9986NEWT:4054NEWT:8654NEWT:8658NEWT:1268063NEWT:8657NEWT:8655NEWT:5147NEWT:98351NEWT:1263854NEWT:118503NEWT:2059687NEWT:160488NEWT:63558NEWT:28104NEWT:207559NEWT:407821NCBITaxon:2NEWT:85057NEWT:985076NEWT:568708NEWT:1215323NEWT:986NEWT:52641NEWT:7159NEWT:267671NEWT:28532NEWT:7160NEWT:353152NEWT:8496NEWT:7165NEWT:1257118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T:1640346NEWT:230741NEWT:8296NEWT:117008NCBITaxon:1313NEWT:167748NEWT:1163394NEWT:44544NEWT:529818NEWT:400540NEWT:4911NEWT:645463NEWT:152480NEWT:129249NEWT:1077286NEWT:943129NEWT:1221594NEWT:990119NEWT:77216NEWT:1524260NEWT:1111708NEWT:227321NEWT:493760NEWT:93163NEWT:281677NEWT:106590NEWT:260710NEWT:257313NEWT:3827NEWT:400772NEWT:128161NEWT:930945NEWT:1162296NEWT:106592NEWT:106350NEWT:46704NEWT:482428NEWT:1187947NEWT:269796NEWT:10312NCBITaxon:1773NEWT:54571NEWT:190304NEWT:412754NEWT:9598NEWT:8030NEWT:1423772NEWT:51056NEWT:53232NEWT:1235987NCBITaxon:2509511NEWT:1639NEWT:188229NEWT:246437NEWT:3818NEWT:4909NEWT:443256NEWT:759272NEWT:432359NEWT:1182263NEWT:943146NEWT:2711NEWT:537457NEWT:479249NEWT:9103NEWT:715877NEWT:862238NEWT:29159NEWT:1155748NEWT:10306NEWT:9595NEWT:8022NEWT:6087NEWT:145943NEWT:595536NEWT:240906NEWT:593117NEWT:89920NEWT:29158NEWT:3633NEWT:319761NEWT:3635NEWT:5811NEWT:44744NEWT:3879NEWT:180923NEWT:1328881NEWT:411483NEWT:76336NEWT:3870NEWT:884019NEWT:198215NEWT:411490NEWT:200298NEWT:169963NEWT:1400870NEWT:587NEWT:588NEWT:585NEWT:343NCBITaxon:43925NEWT:84005NEWT:225117NEWT:746128NEWT:8090NEWT:5801NEWT:43665NEWT:212035NEWT:5802NCBITaxon:1653394NEWT:1266464NEWT:1205NEWT:214693NEWT:367830NEWT:178616NEWT:95145NEWT:358NEWT:375451NEWT:357NEWT:9172NEWT:2770979NEWT:2010198NEWT:1277351NEWT:1594786NEWT:457428NEWT:106178NEWT:411469NEWT:84023NEWT:1679NEWT:559292NEWT:214684NEWT:569NEWT:411464NEWT:411460NEWT:2762NEWT:1174673NEWT:1328426NEWT:562NEWT:28035NEWT:411470NEWT:1313166NEWT:2094720NCBITaxon:2697049NCBITaxon:138NEWT:28038NEWT:294750NEWT:1663NEWT:62141NEWT:1422NEWT:1423NEWT:4932NEWT:3603NEWT:2756NEWT:2759NEWT:3848NEWT:3847NEWT:38293NEWT:1428NEWT:198466NEWT:339NEWT:327159NEWT:178876NEWT:1660NEWT:2751NEWT:327160NEWT:573NEWT:571NEWT:142479NEWT:12922NEWT:163159NEWT:29595NEWT:1306413NEWT:241368NEWT:2813599NEWT:518766NEWT:42528NEWT:2664373NEWT:190802NEWT:1213463NEWT:9417NEWT:1216979NEWT:7237NEWT:264483NEWT:5059NEWT:509924NEWT:28450NEWT:115104NEWT:1121114NEWT:5061NEWT:663NEWT:8570NEWT:127569NEWT:1081927NEWT:1238993NEWT:1408434NEWT:67825NEWT:187878NEWT:1241935NEWT:220668NEWT:1821314NEWT:1249668NEWT:426430NEWT:9407NEWT:37181NEWT:9646NEWT:5286NEWT:7227NEWT:7469NEWT:9402NEWT:9644NEWT:415540NEWT:336722NEWT:675060NEWT:416870NEWT:334542NEWT:313625NEWT:51240NEWT:27592NEWT:5052NEWT:426428NEWT:227289NEWT:63459NEWT:2652157NEWT:2652158NEWT:1276815NEWT:40324NEWT:588858NEWT:9638NEWT:9639NCBITaxon:11071NEWT:242231NEWT:197221NEWT:7215NEWT:528354NEWT:51453NEWT:2172103NCBITaxon:59201NEWT:7460NEWT:504457NEWT:434271NEWT:392227NEWT:747679NEWT:66712NEWT:42752NEWT:103372NEWT:746360NEWT:862751NEWT:89764NEWT:470150NEWT:9627NEWT:543734NEWT:1009714NEWT:73239NEWT:38018NEWT:226186NEWT:9870NEWT:8782NEWT:30538NEWT:115357NEWT:435590NEWT:1902NEWT:63436NEWT:1908NEWT:1169740NEWT:13164NEWT:1601067NEWT:216129NEWT:1245012NEWT:2664179NEWT:623NEWT:7038NEWT:6192NCBITaxon:1639NCBITaxon:38727NEWT:74790NEWT:2829NEWT:1428006NEWT:62335NEWT:366581NEWT:757417NEWT:216599NEWT:405534NEWT:216595NEWT:40559NEWT:51208NEWT:216597NEWT:9689NEWT:8355NEWT:633NEWT:1607839NEWT:9685NEWT:7029NEWT:6182NEWT:1283300NEWT:9694NEWT:1218707NEWT:186990NEWT:8364NEWT:632NEWT:1204551NEWT:6183NEWT:99586NEWT:630NCBITaxon:11019NEWT:334747NEWT:3907NEWT:61235NEWT:392021NEWT:343691NEWT:1457551NEWT:15368NEWT:1491466NEWT:436486NEWT:340188NEWT:51655NEWT:1933298NEWT:170187NEWT:260707NEWT:121845NEWT:8595NEWT:7263NEWT:186763NEWT:185431NEWT:216591NEWT:29760NEWT:260704NEWT:1719NEWT:703612NEWT:260705NEWT:1202347NEWT:692672NEWT:2697049NEWT:1505907NEWT:1216981NEWT:1109443NEWT:1249885NEWT:7004NEWT:1972855NEWT:9669NCBITaxon:6073NEWT:1089448NEWT:1415167NEWT:1329349NEWT:607699NEWT:6161NEWT:208963NEWT:208964NEWT:27337NEWT:575584NEWT:181088NEWT:48498NEWT:185687NEWT:3039NEWT:669202NEWT:2706014NEWT:8727NEWT:8726NEWT:6306NEWT:10090NEWT:8729NEWT:4120NEWT:8720NEWT:5693NEWT:2186NEWT:8724NEWT:8723NEWT:283166NEWT:37334NEWT:5697NEWT:6544NCBITaxon:10357NCBITaxon:10114NEWT:2190NCBITaxon:10359NEWT:1685931NEWT:1203258NEWT:242619NEWT:49317NEWT:557723NEWT:557722NEWT:160791NEWT:1353008NEWT:34073NEWT:696811NEWT:402042NEWT:9808NEWT:373995NEWT:5689NEWT:8715NEWT:839NEWT:544404NEWT:92519NEWT:2173NEWT:943925NEWT:4111NEWT:37349NEWT:4113NEWT:837NEWT:640459NEWT:72568NEWT:36015NEWT:51641NEWT:1232801NEWT:81866NEWT:1262469NEWT:5691NEWT:48479NEWT:61435NEWT:627025NEWT:1097677NEWT:375939NEWT:61674NEWT:2072580NEWT:669465NEWT:3258NEWT:1117957NEWT:6526NEWT:112018NEWT:8709NEWT:4100NEWT:1159556NEWT:5671NEWT:1076NEWT:4102NEWT:4101NEWT:6763NEWT:8700NEWT:803NEWT:29722NEWT:1692259NEWT:1486917NEWT:419947NEWT:2847306NEWT:468911NEWT:13373NEWT:817NEWT:7604NEWT:6756NEWT:818NEWT:135588NEWT:1843183NEWT:5661NEWT:157592NEWT:1061NEWT:51866NEWT:29719NEWT:58002NEWT:13136NEWT:5665NEWT:6754NEWT:1179777NEWT:6997NEWT:452589NEWT:509068NEWT:5666NEWT:4577NEWT:813NEWT:5664NEWT:2157NEWT:146479NEWT:1042986NEWT:1911079NEWT:69373NEWT:1843192NEWT:206189NEWT:427577NEWT:264203NEWT:1480154NEWT:9859NEWT:1274414NEWT:9612NEWT:38626NEWT:1286640NEWT:38865NEWT:3077NEWT:9615NEWT:1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