Tandemly Duplicated Rubisco Activase Genes of Cereals Show Differential Evolution and Response to Heat Stress

Tandemly Duplicated Rubisco Activase Genes of Cereals Show Differential Evolution and Response to Heat Stress | 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 Tandemly Duplicated Rubisco Activase Genes of Cereals Show Differential Evolution and Response to Heat Stress Ragupathi Nagarajan, Kaviraj Singh Kahlon, Amita Mohan, Kulvinder S. Gill This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4676428/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 21 Dec, 2024 Read the published version in Plant Molecular Biology → Version 1 posted 6 You are reading this latest preprint version Abstract Heat stress affects various components of photosynthetic machinery of which Rubisco activation inhibition due to heat sensitive Rubisco activase (RCA) is the most prominent. Detailed comparison of RCA coding genes identified a tandem duplication event in the grass family lineage where the duplicated genes showed very different evolutionary pattern. One of the two genes showed high level of sequence conservation whereas the second copy, although present only 1.5kb away, was highly variable among various plant species because of loss of introns, alternative splicing and loss of the last exon coding redox regulated C-terminal extension domain. Gene specific expression analysis, both at the transcription as well as the protein level, showed very different expression pattern of the two RCA copies. Expression of the highly conserved copy was higher under normal plant growing conditions that decreased many folds under heat stress with substantial genotypic variation, but the variable copy showed much higher expression under heat stress conditions across all grass species. The cultivated rice has only one functional gene as the second copy became nonfunctional due to multiple deletions but Oryza brachyantha and Oryza australiensis still have two functional Rca genes. Detailed analysis of the promoter region of the two copies among various plant species showed insertion of several transposable elements harboring heat responsive elements in the heat inducible copy of the gene. The conserved RCA copy of wheat didn’t have any transposable insertions whereas in that of maize has one heat shock element and sorghum had two. It would be interesting to study if the higher level of heat stress tolerance observed in sorghum and maize is associated with the differences observed for RCA. Rubisco Activase Cereal Crops Gene Duplication Heat Stress Photosynthesis Molecular Evolution and Regulation of Gene Expression Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction The optimal temperature for photosynthesis varies among crops, from 25-30°C in wheat to 30-35°C in rice (Nagai and Makino, 2009 ). Every degree-Celsius increase over the optimal temperature may result in up to 6% reduction in grain yield of wheat and 3.2% of rice (Zhao et al., 2017 ). Although many plant metabolic pathways are affected by heat stress, the effect on photosynthesis is critical as any constraint on the process will directly impact development, growth, biomass production, and crop yield. Plant photosynthetic machinery involves complex components and processes, many of which including Rubisco activation, enzymes and proteins of photosystems, membrane permeability and ATP generation are vulnerable to high temperature (Allakhverdiev et al. 2008 ). The critical temperature and its duration affecting each photosynthetic process often differ among species (Feller et al., 1998 ). Plants exposed to temperatures above 40°C for extended periods show irreversible damage to the permeability of thylakoid membranes and electron transport chain, resulting in photosynthesis inhibition. The photosystems and electron transport are usually not affected until 40°C, whereas for many species including some important crop plants including wheat the Rubisco activation level and photosynthetic rate (Pn) start to decline even at moderately elevated temperatures (Feller et al., 1998 ). The functional state of Rubisco is maintained by Rubisco activase (RCA) that removes inhibitory sugar phosphates from the active sites of Rubisco with energy derived from ATP hydrolysis (Salvucci et al., 1985 ). RCA is a nuclear encoded, cytosol synthesized chloroplast protein, generally consisted of two immunologically related polypeptides called α isoform (45 to 46 kDa) and β isoform (41 to 43 kDa). The size difference between the two isoforms is due to the presence of a redox regulated C-terminal extension (CTE) domain that is present in the α isoform but is lacking in the β form. In most plants, both α and β isoforms of RCA are capable of activating Rubisco (Werneke et al., 1989 ; Rundle and Zielinski, 1991 ; To et al., 1999 ; Salvucci et al., 2003 ). Shown in Arabidopsis, the two cysteine (Cys) residues located in the CTE domain of RCA α isoform make a disulfide bond under oxidizing conditions. This significantly increases its sensitivity to physiological level of ADP/ATP ratio and affects both ATP hydrolysis and Rubisco activation (Zhang and Portis, 1999 ; Zhang et al., 2002 ). The major function of larger isoform is to regulate the activity of smaller RCA isoforms under low light/dark conditions in order to conserve energy. The equimolar concentrations of the larger and the smaller isoforms, completely inhibited the activity of smaller isoform under higher ADP/ATP ratio in light conditions (Zhang et al., 2001 ). Interestingly, tobacco, which expresses only the β isoform, is also sensitive to physiological levels of ADP/ATP ratio and responds similar to species with both isoforms (Carmo-Silva and Salvucci, 2013 ). Selection for genotypes with higher Rca expression levels at normal conditions in soybean, maize, and some other crops showed a positive correlation with Rubisco activity, photosynthetic rate, and seed yield (Morales et al., 1999 ; Yin et al., 2010 , 2014 ). In-vitro analysis has shown that while the RUBISCO enzyme is stable even at 50 o C, RCA is very sensitive to heat stress and its activity start to decline at temperatures over 35 o C (Crafts-Brandner and Salvucci, 2000 ). As a result, at higher temperatures RCA is unable to keep pace with faster Rubisco deactivation thus constrains photosynthesis (Crafts-Brandner and Salvucci, 2000 ). In crops including cotton and wheat, even a moderate level of heat stress decreases the Rubisco activation levels, which correlated with reduced net photosynthesis, increased RuBP level, and decreased levels of 3-phosphoglycerate in leaves (Law and Crafts-Brandner, 1999 ). Independent studies on various crops have shown that photosynthetic heat acclimation is directly associated with the acclimation of heat sensitive RCA to high temperature (Law and Crafts-Brandner, 1999 ; Yamori et al., 2006 ). This conclusion was further supported by the studies showing increase in the thermal tolerance of Arabidopsis photosynthesis by improving RCA thermostability (Kurek et al., 2007 ; Kumar et al., 2009 ). Previously, we showed that the Rca gene structure is highly conserved in higher plants, and the common grass ancestor Rca , similar to Arabidopsis AtRca1 , contained seven exons and six introns, enabling expression of the RCA α isoform with redox regulated CTE domain coded from the seventh exon (Nagarajan and Gill, 2018 ). The alternatively spliced mRNA of the same gene contains a premature stop codon and produces a shorter redox insensitive RCA β isoform (Werneke et al., 1989 ). We also reported that the Rca gene is tandemly duplicated ( Rca1 and Rca2 ) among grass species and the duplication event probably occurred in the common grass ancestor, but before the divergence of subfamilies (Nagarajan and Gill, 2018 ). Heat stress induced changes in the RCA expression pattern at protein level were reported earlier in rice (Scafaro et al., 2010 ; Wang et al., 2010 ), wheat (Feller et al., 1998 ; Law and Crafts-Brandner, 2001 ; Ristic et al., 2009 ), and maize (Crafts-Brandner and Salvucci, 2002 ; Ristic et al., 2009 ). High temperature stress altered the ratio of α and β isoform levels in rice (Wang et al., 2010 ), but induced new RCA isoform expression in wheat and maize (Law and Crafts-Brandner, 2001 ; Crafts-Brandner and Salvucci, 2002 ; Ristic et al., 2009 ). A recent study in wheat showed that heat stress altered the expression pattern of Rca genes (Scafaro et al., 2019 ). Later, (Degen et al. , 2020) showed a single amino acid substitution (M159I) in TaRCA2-β isoform could increase the thermal stability of the enzymatic activity by up to 5°C. However, the heat stress induced expression pattern of RCA isoforms in important cereal crops and key evolutionary changes causing the differential gene expression are still not well studied. Earlier, we proposed an evolutionary model for tandemly duplicated Rca genes of the grass species (Nagarajan and Gill, 2018 ). We also showed that after the duplication event, one of the gene copies, Rca1 , have lost the third, the fifth and the sixth introns, which also abolished the alternative splicing mechanism of the Rca transcripts. Other changes observed in Rca1 among various grass species included nonfunctional Rca1 in some Oryzoideae species; 2) loss of Rca1 sequences coding the redox regulated CTE domain in Pooideae including that in Brachypodium, barley, and wheat and the loss of various introns (Ragarajan and Gill, 2018). 3) loss of Rca2 sequences coding for the redox regulated CTE domain in Panicoideae species (maize, sorghum and foxtail millet), and 4) loss of various introns in both Rca1 and Rca2 of these crops. Whereas, the second gene copy, Rca2 , did not amass any major changes in its gene structure although loss of some introns and sequences coding for the redox regulated CTE domain in Panicoideae species including maize, sorghum and foxtail millet was observed. In the present study, first, we studied the origin of the tandem duplication of Rca genes. Specifically, we tracked the evolution of the rice OsRca1 by comparing the sequences of various Oryzoideae species. Second, we studied the effect of heat stress on the expression pattern of tandemly duplicated cereal Rca gene copies both at the mRNA as well as at the protein level and investigated possible causes (i.e. promoter elements) of the variation. Finally, we performed a phylogenetic study to analyze the rate of amino acid substitution and the evolutionary relationship of different monocot RCA isoforms. The information was then used to understand the role of tandemly duplicated Rca genes in Rubisco activation, and photosynthetic acclimation to heat stress in important cereal crops. Results Tandem duplication of Rca gene occurred after the divergence of Poales families With the objective to accurately determine origin of the tandem duplication in the RCA gene, we evaluated the Rca gene structure in three distantly related monocot species: banana ( Musa acuminata ), date palm ( Phoenix dactylifera ) and pineapple ( Ananus comosus; family: Bromeliaceae; order Poales). Unlike the grass family, no tandem duplication of the Rca gene was observed in any of these species. Instead, these species contained three Rca gene copies on separate chromosomes (table S1). The Rca gene copies in banana were located on chromosomes 2, 10 and 11, whereas in date palm, the copies were present on two different unplaced scaffolds NW_008246537.1 (1.6 Mbp) and NW_008246711.1 (0.506 Mbp) and were not tandemly duplicated as was seen in grasses. The gene copies in pineapple were located in the linkage groups (LG) LG14, LG18, and LG23. Otherwise, the structure of the gene copies was similar to those of other higher plants. In all three species, the exon sizes and relative order were highly conserved among the three copies as was seen in other plant species. Major variation was however observed for intron length (Fig. 2 b). Based on these observations, we concluded that the gene duplication event occurred after the divergence of the Poales families and is probably specific to the Poaceae. Rca gene structure among cereals After evaluating Rca gene structure among various grass family members, we identified and sequence confirmed Rca gene copies from seven ( O. brachyantha , O. australiensis , O. sativa , T. aestivum , Z. mays , S. bicolor , and Setaria italica ) species. Since rice showed major variation for the Rca gene structure, we also obtained sequence of seven cultivated rice accessions and two wild rice species (Oryza australiensis and Oryza brachyantha ). For each accession, both genomic and cDNA copies were cloned and sequenced. While the Rca2 sequence from one of the accessions was reported earlier (To et al., 1999 ; Scafaro et al., 2016 ), we cloned and sequenced the Rca1 gene from O. australiensis (GenBank Acc. MH115971), and O. brachyantha (GenBank Acc. MH220418), and Rca2 gene from O. brachyantha (GenBank Acc. MH660917). The structural comparison of Rca1 gene copy across various cultivated and wild rice species revealed presence of large deletions in the exons 2, 3, 4, 5, 6 and 7 which appeared to have caused frameshift mutation in the Rca1 of cultivated rice species (Fig. 3 ). In other species, between 11 to 13 independent deletions were detected in all four exons, resulting in 157 to a total loss of 218 nucleotides. These Rca1 deletions appear to have occurred multiple times, but in different time periods during the evolution. For example, O. barthi and O. glumipatula have a 15 bp deletion on the third exon, whereas O. nivara , O. sativa , and O. rufipogon showed a 39 bp deletion (Fig. 3 ; Box1). Similarly, the fourth exon of Rca1 of O. nivara , O. sativa , and O. rufipogon contains two additional deletions of 13 and 24 nucleotides (Fig. 3 ; Box 2), which were absent in other wild species. Additionally, species specific indels in the Rca1 gene were also identified. The six bp addition in the second exon of O. punctata Rca1 , a 25 bp deletion in the fourth exon of O. nivara , and a nine bp instead of the 14 bp deletion in the second exon of O. glumipatula Rca1 occurred probably after the divergence of the species. However, it is not clear how the 24 bp deletion is absent in O. sativa ssp. indica but present in most other species including O. sativa ssp. japonica . Interestingly, the intron lengths of Rca1 were determined before the major Oryza species divergence and maintained thereafter (Fig. 3 ). Alignment of RCA2 amino acid sequences of these species showed 100% similarity, except for O. brachyantha and O. punctata , which had only 95.5% and 97.4% similarities with the other Oryza species respectively. Rca1 gene structure in Oryzoideae subfamily From the comparative analysis of Rca genes in Poaceae, it was evident that both Pooideae and Panicoideae subfamilies have two functional Rca gene copies present in tandem (Fig. 1 ). The OsRca1 copy contained multiple small deletions that seem to have occurred after the divergence of these sub-families. Comparison of the two wild and seven cultivated rice species revealed the presence of a putative Rca1 gene in all these species. Structurally, the Rca1 gene in O. australiensis , O. brachyantha , and O. punctata contained four exons with slight size variation for the three introns (Fig. 3 ). The remaining five species including the two accessions of cultivated rice showed length of the three introns were remarkably conserved. The exons of these five species showed 11 to 13 deletions (Fig. 3 ). Nine of these deletions were common to all five species and the remaining were specific to one or more species. At the predicted protein level, the largest protein among these three species was predicted to be 466aa in O. punctata . The predicted protein in O. australiensis is 465aa as compared to 458 in O. brachyantha . Unlike the RCA1 of Pooideae members, these predicted peptides also contain the light sensitive CTE domain as identified in the RCA1 of the Panicoideae members. Thermostability conferring amino acids in RCA1 isoform of cereals Previous studies showed that Arabidopsis transgenic plants expressing RCA variants with certain amino acid substitutions were heat tolerant, and the modified Arabidopsis RCA showed up to 10°C increase in stability and Rubisco activation activity (Kurek et al., 2007 ). One such substitution T274R (Threonine 274 Arginine) in the variant 183H12, which alone could provide the maximal heat stability and activity, was identified in all three copies of TaRCA1 and ZmRCA2 (Fig. 6 ). Other natural variants observed in the same position were glutamine (Q) in SbRCA1, SbRCA2, and ZmRCA1, and lysine (K) in ObRCA1 and ObRCA2 of O. brachyantha , and in all of the RCA2 isoforms of rice and wheat. Based on these results, we hypothesize that the isoform expressed during heat stress may have better thermal stability than the isoforms expressed at normal conditions because of the putative thermal tolerant amino acid substitutions. The role of these natural RCA variants in thermal tolerance is not known, and further experiments are needed to identify the significance of these sequence changes in providing thermal tolerance. Phylogenetic analysis of monocot RCA Phylogenetic analysis performed with the alignment of RCA protein from distantly related monocots, using the MEGA7 maximum likelihood method, revealed different clusters of RCAs, based on the sequence similarity and evolutionary pattern (Fig. 7 ). In deep nodes, the phylogenetic relationship is unclear, and the bootstrap (BS) values are low (< 70%). The comparative analysis showed how the duplicated Rca genes evolved in cereals. Thus, we did not collapse the deep nodes with lower statistical support. Based on the phylogenetic analysis, RCA isoforms of banana, date palm and pineapple, and the cereal isoforms, formed distinct clades, with cereals further comprising of RCA1 and RCA2 from the tandemly duplicated genes. Although the RCA isoforms of banana (MaRCA1, 2 and 3), date palm (PdRCA1 and 2), and pineapple (AcRCA1, 2 and 3) were clustered together, the subgroup pattern within the cluster suggests that the genes coding these isoforms were duplicated or triplicated after the divergence of monocot orders. Similarly, the RCA1 and RCA2 of cereals formed different clades, but within each clade, the isoforms of closely related species, or species from a shared family, showed less divergence with high BS values. The branch and individual leaf lengths show the rate of substitution, the RCA1 isoforms of wild rice and Panicoideae species showed a higher rate of changes than did the RCA1 of Pooideae. Together, these results suggest that after the divergence for a common monocot ancestor, the Rca gene copy number increased either through polyploidization or tandem duplication and evolved at different rates in the studied species. Expression pattern of RCA isoforms in cereals Detailed analysis of 302 ESTs from banana ( Musa acuminata ) and 331 from pineapple ( Ananus comosus ) showed that in both banana and pineapple, only one of the three copies codes for RCA-α, whereas the other two copies code for only the β isoform (Supplementary Table S1). EST sequences showed no evidence of alternative splicing in the RCA-α coding copy, suggesting a possible loss of the splice junction during evolution. In comparison, alternate splicing was observed in date palm ( Phoenix dactylifera) based on the analysis of 305 ESTs. One of the two Rca copies, PdRca1 , codes for both RCA-α and RCA-β isoforms via alternative splicing, whereas the other gene copy ( PdRca2 ) codes only for the RCA-α isoform. Evaluating the EST data, MaRca1-α, MaRca2-β , and MaRca3-β of Banana has 101, 101 and 100 ESTs respectively; PdRca1-α, PdRca1-β and PdRca2-α of date palm has 103, 102 and 100 ESTs respectively and AcRca1-α, AcRca2-β and AcRca3-β of pineapple has 110, 115 and 106 ESTs respectively. Based on this data, the three splice variants appear to be equally expressed in these three plants. With the objective to compare expression of the gene(s) at the RNA level with that at the protein level, and to identify the relative contribution of the two tandemly duplicated Rca gene copies, we performed gene specific expression analyses using quantitative Real Time PCR (qRT-PCR) for both Rca1 and Rca2 genes of O. brachyantha , O. australiensis , O. sativa , T. aestivum , Z. mays , and S. bicolor at normal as well as at high temperature conditions (Fig. 5 ). In cultivated rice ( O. sativa ), either under heat stress or normal plant growth conditions, no expression was observed for the Rca1 copy in either of the two cultivars that were used for the analysis (Fig. 5 a). O. australiensis showed some expression of Rca1 under normal growth conditions and the expression reduced both under 35 o C and 45 o C treatment. O. brachyantha on the other hand showed a different expression pattern for Rca1 where its expression was very low under normal plant growth conditions but increased under the heat stress conditions. Rca1 expression was however significantly less than that for Rca2 . The Rca2 copy in cultivated rice showed very high expression level under normal plant growth conditions and its expression reduced dramatically under heat stress conditions. There was significant difference between the two cultivated rice lines for the expression of Rca2 copy. The two rice wild relatives showed dramatically higher expression for Rca2 under heat stress. Even between the two wild relatives, O. brachyantha showed significantly higher expression at 45 o C treatment as compared to O. australiensis . Heat stress also affected the alternative splicing mechanism, as the reduction in the levels of transcripts coding RCA2α varied during heat stress (Fig. 4 ). In rice, OsRca2 gene expresses OsRca2α and OsRca2β transcripts via alternate splicing mechanism. At normal conditions, the level of OsRca2β transcripts was many hundred-folds higher than that of the OsRca2α transcripts, but at 42°C, the OsRca2α transcript level was either equal or higher than that of the OsRca2β transcripts. Interestingly, ObRca2α transcripts were stable at all studied temperatures, however, ObRca2β showed more than two-fold reduction at higher temperatures. These results suggest that the temperature increase, at least among various rice species, favors transcripts coding RCA2α isoform. Even though the transcripts encoding OsRCA2α and OsRCA2β showed a significant reduction at 45°C in rice, the quantity of both the isoforms from the heat stressed leaves is comparable or higher to their control leaves (Fig. 4 ). The expression pattern of the Rca gene copies in wheat was very different from that observed in rice. Rca1 showed essentially no expression at normal plant growth conditions, but its expression increased dramatically under heat stress conditions (Fig. 4 b). The expression pattern was very similar among the three cultivars (Chinese Spring, Giza168 and PBW343) although significant differences were observed among cultivars. In all three cultivars, transcript level for Rca2 copy was significant at all three temperature conditions although its level at 37 o C was significantly higher than either of the other two conditions. Genotypic differences were significant among the three genotypes at each temperature condition. Between the two isoforms of Rca2 , transcript level of Rca2α isoform was much lower than the Rca2β form and stayed about the same over the three temperature treatments (Fig. 4 b). The Rca2β form showed significant increase at 37 o C treatment but reduction at 42 o C. Both in maize and sorghum, Rca1 copy made only the alpha isoform of the gene whereas the Rca2 copy made the beta form (Fig. 4 c and d). In both of these species, the beta form was made at all three temperatures although significant differences were observed both among temperatures as well as among genotypes. The alpha form showed essentially no accumulation at 25 o C, slight increase at 35 o C treatment and dramatic increase at 45 o C. In general, the genotypic differences for relative expression of the two isoforms were much higher than that observed among the wheat genotypes. Using the same plant samples as that used for real-time expression analysis, RCA protein level was also studied using immunoblot analysis with an RCA specific antibody (Fig. 5 ). Except for wheat, all of the studied cereals expressed a 42 kDa RCAβ isoform at normal as well as under heat stress conditions whereas the 46 kDa RCAα isoform was expressed only during the heat stress conditions (Fig. 5 ). In comparison to Rubisco, the expression level of wheat RCAβ was significantly higher at the normal temperature as compared to that under heat stress. Relative to RCAβ, expression of the RCAα is much lower at normal conditions but is undetectable after the heat treatment. Although it is difficult to accurately quantify, expression levels of both Rubisco, as well as RCA of rice, maize, and sorghum, increased after 45°C treatment as compared to that at the normal temperature. Wheat, on the other hand did not show the 46 kDa RCAα isoform under heat stress conditions along with a significant reduction in the RCAβ level. With the current analysis, it was not possible to determine if the 42 kDa RCAβ isoform of wheat is expressed from TaRca1 or the TaRca2 copy. Regulatory elements in the Rca genes of cereals As the two Rca gene copies of cereals showed very different expression pattern, we looked for differences in expression control elements between promoters of the two copies. We evaluated regulatory elements in ~ 1.5 kb region upstream of the transcription start site of the two Rca gene copies of the studies species along with the intronic sequences, with a focus on heat regulated elements (material and methods). Along with the core promoter elements, this analysis showed various putative cis-elements including that for circadian rhythm, light, hormone and biotic and abiotic stress responsives (Table S2). The Rca1 copy showed insertion of heat shock elements (HSE) in its promoter with wild rice ( O. brachyantha ) having three HSEs. These HSEs were missing in the Rca1 promoter of cultivated rice ( O. sativa ). Interestingly, Rca2 of O. sativa have HSEs in its promoter and in the 5th intron. Transposable element (TE) of size 366bp, 347bp and 320 bp were present in the promoter of Rca1 , promoter of Rca2 and in the 5th intron of Rca2 , respectively (Fig. 1 ). In wheat, Rca1 promoter of the A and the B genome showed one HSEs and the D genome has two. There were no HSEs present in the promoter of Rca2 of wheat but promoter of D genome showed insertion of 162 bp TE. In maize ( Z. mays ) and sorghum ( S. bicolor ), both Rca1 and Rca2 had insertion of HSEs with sorghum having maximum of 5 HSEs in the promoter of Rca1 . Promoter of Rca2 had two HSEs in sorghum and one in maize. TE were also present in the promoter of Rca1 in both maize and sorghum (Fig. 1 ) Discussion Adaptation of organisms to the changing environmental conditions via modifications in the basic biological processes is a common feature of evolution. For better adaptation, an organism often relies on new gene mutations which may develop neo-functions, however, that could also cause detrimental effects if the mutated gene is essential for the organism’s survival. Gene and genome duplications provide the base genetic material for the origin of novel genes or trait development during evolution. Tandem duplication of Rca genes was first reported in barley (Rundle and Zielinski, 1991) and later in wheat (Carmo-Silva et al. , 2015). In rice, DNA gel blot analysis with cDNA coding RCA as a probe showed the presence of more than one Rca gene in the genome (Zhang and Komatsu, 2000). We (Nagarajan and Gill, 2018) showed that the tandem duplication event occurred in the common grass ancestor and the structural changes then accumulated after the divergence of the Poaceae families. In this study we have shown that this tandem duplication of Rca gene is Poaceae-specific and probably occurred after the divergence of Poales order as one (Bromeliaceae) of the 16 families did not have the duplication. It would be possible to pinpoint the exact timeline of the duplication event once the genome sequence information for the other 14 families is available. Phylogenetic and fossil data analyses have suggested that the grass sub-families probably diverged from a common ancestor between 50-70 million years ago (MYA) (Kellogg, 2001; Salse et al. , 2008). The tandem duplication event of Rca genes might have occurred before the divergence of the grass sub-families as the Oryzoideae, Pooideae, and Panicoideae members have the duplicated genes (Fig. 2). The conserved heat induced expression pattern of Rca1 in all three sub-families suggests that the promoter elements required for heat responsive expression might have integrated into the Rca1 promoter of the grass ancestor. Later, species specific TE insertions added additional HSEs in the promoter region and further altered the gene expression. Effect of TE insertions on the neighboring gene expression is a well-documented phenomenon in plants and other organisms (see reviews, Feschotte, 2008; Lisch, 2013; De Souza et al., 2013). Similarly, integration of biotic and abiotic responsive cis-regulatory elements in the promoter regions of genes by TE was also reported in plants (Beguiristain et al. , 2001; Naito et al. , 2009), including HSEs (Ito et al. , 2013). The insertions of TEs with HSE elements in duplicated Rca gene regions of cereals could be the source for evolutionary adaptation through altered gene expression. Heat stress induced changes in the RCA expression pattern were reported earlier in rice (Scafaro et al. , 2010; Wang et al. , 2010), wheat (Feller et al. , 1998; Law and Crafts-Brandner, 2001; Ristic et al. , 2009), and maize (Crafts-Brandner and Salvucci, 2002; Ristic et al. , 2009). Our copy specific Rca transcript quantification along with immunoblot analysis have specifically revealed expression pattern of each of the Rca copies and of its splice varients. We have also shown differences and similarlities among Rca of various species for a differential response to heat stress. In comparison to maize and sorghum, the wheat Rca showed almost an opposite accumulation pattern of different Rca isoforms and splice varients (Figures 4&5). Role of these differences in explaining differences in heat stress tolerance among wheat, maize and sorghum needs to be validated. One of the important findings of this study is the identification of a functional Rca1 gene copy among wild rice species. The genus Oryza is composed of 23 recognized species, including two cultivated ( O. sativa and O. glaberrima ) and 21 wild rice species with 10 different genomes (AA, BB, CC, BBCC, CCDD, EE, FF, GG, HHJJ, and HHKK) (Ge et al. , 1999; Vaughan et al. , 2003). Our analysis on the non-functionalization of Rca1 showed that the species O. sativa , O. glaberrima , O. barthii , O. glumipatula , O. nivara , and O. rufipogon with AA genome have multiple deletions in the exonic sequences thus have non-functional Rca1 . Interestingly, O. punctata with BB genome, O. australiensis with EE, and O. brachyantha with FF genome contain a complete and functional Rca1 gene. These results suggest that the non-functionalization of Rca1 in the AA genome species occurred after the divergence of Oryza species and also specifically in the AA genome lineage. Recent research on thermal tolerance of wild rice species O. australiensis showed unaffected net photosynthetic and leaf elongation rates, even at 45°C, whereas O. meridionalis and cultivated rice O. sativa showed intermediate tolerance to > 50% decline in net photosynthesis and leaf growth rates (Scafaro et al. , 2016). Rubisco activation levels of these species at high temperature correlated with the net photosynthesis and leaf elongation rate. Interestingly, the thermal stability of recombinant OaRCA2 was similar to the OsRCA2 of cultivated rice in the in vitro assays, but the leaf extracted RCA of O. australiensis showed no Rubisco activation inhibition at temperatures as high as 42°C, whereas rice RCA inhibition occurred at temperatures above 34°C. The authors characterized the OaRca2 of O. australiensis and compared its thermal stability with that of rice OsRca2 . They speculated that some unknown factor(s) in the leaf extract could contribute to its higher thermal activity and stability (Scafaro et al. , 2016). Based on our Rca1 comparative analysis of Oryza sp. and heat stress induced expression of OaRca1 of O. australiensis , we propose that the presence of OaRCA1 could be one of the reasons for increased thermal stability of Rubisco activation and unaffected net photosynthetic observed in the above-mentioned study. However, further experiments are needed to characterize the OaRca1 and utilize this gene in rice breeding through alien introgression to improve photosynthetic thermal tolerance of rice cultivars. Various heat shock elements (HSEs) present in the sequences upstream of the transcription start sites (TSS) were identified in ObRca1 of O. brachyantha , OsRca2 of rice, all three copies of wheat TaRca1 , both ZmRca1 and ZmRca2 of maize, and both SbRca1 and SbRca2 of sorghum (Fig. 1). The number of HSEs also varied among species with the promoter of SbRca1 containing up to five, ObRca1 containing three, TaRca1-D and SbRca2 containing two, and the remaining ones showing only a single HSE in the upstream region. It is tempting to speculate a correlation of higher level of heat stress tolerance in sorghum and wild rice with higher number of HSE’s but additional experimentation is needed to confirm. Previous studies have shown that certain transposable elements are preferentially inserted into the regulatory sequences of various genes and play an important role in the expression control of the neighboring genes (Makarevitch et al. , 2015). In some cases, heat stress induced gene expression was acquired from TE insertion events (Ito et al. , 2011). The duplicated Rca genes of closely related yet evolutionarily diverged cereals, and their heat stress induced expression pattern may serve as an ideal model for understanding different TE insertions and their role in the stress regulated expression. To identify TEs in the genic region, we used the duplicated gene sequences, along with 2Kb sequence flanking on both sides, as the query to search the crop specific TE database. We identified independent TE insertion events in the promoter sequence of all six Rca1 copies of the studied species (Table 2). Similarly, there were TE insertions in the promoter and the fifth intron of rice OsRca2 , and in the promoter of TaRca2-D copy (Fig. 1). Interestingly, many of these TEs contain HSEs; these TEs inserted in different time periods of evolution served as a potential source for the HSEs and may have caused heat stress induced expression pattern in the Rca1 genes. A detailed list of the identified TEs, their sizes, and locations relative to the genes is given in Table 2. In conclusion, the comparative and expression analyses of RCA coding genes of important cereal crops revealed a tandem gene duplication, which occurred in the common grass ancestor subsequently led to neo-functionalization and evolved to respond differently during heat stress. One of the two copies of the tandem duplication maintained a very high level of conservation whereas the second copy showed tremendous divergence to evolve species specific function of the gene. We identified the insertion of various transposable elements harboring heat responsive elements in the promoter region. These results suggest that one of the duplicated Rca genes showing divergence, which could be responsible for the heat induced expression pattern and thermal stability of these heat tolerant cereal crops. Understanding these evolutionary changes are important as this knowledge can be used to engineer future plants with better and wider adaptability. Materials and Methods Gene model prediction and validation The genomic regions comprising Rca genes of banana, date palm, and pineapple were searched using the well characterized 474 aa sequence of AtRCA1 (AT2G39730.1) ( https://www.arabidopsis.org/ ) as the query in Phytozome 11 ( https://phytozome.jgi.doe.gov ) and NCBI database. We selected banana, date palm, and pineapple because of its position in the monocot evolution with sequenced genome at the time of analysis. Gene models were developed using the predicted transcripts for each of these genes from NCBI database (Table S1). The Rca gene models for rice (cv. Nipponbare), wheat (cv. Chinese spring), maize (inbred B73), and sorghum (inbred BTx623) were adopted from our previous study (Nagarajan and Gill, 2018 ). To identify the Rca genes of wild rice species, AtRCA1 was used as a query sequence to search 11 Oryza species (Jacquemin et al., 2013 ) of Ensembl plants DNA database with default settings ( http://plants.ensembl.org/index.html ). For the Oryza Rca1 structural analysis, we did not include O. meridionalis , O. glaberrima , and O. longistaminata , because the sequences for the region was not complete in these three species. The criteria used for searching true orthologs in this study were described elsewhere (Dhaliwal et al., 2014 ). For identification of cis-regulatory elements in the promoter region, we used the 1.5 Kb upstream sequence from ATG start codon for each gene to search in PlantCARE database (Lescot et al., 2002 ). The transposable elements for Oryza species were identified from RiTE-DB ( http://www.genome.arizona.edu/cgi-bin/rite/index.cgi ); wheat from cereal repeat database ( http://wheat.pw.usda.gov/ggpages/Repeats/ ), maize from MaizeGDB ( http://www.maizegdb.org ), and sorghum from TREP database ( http://botserv2.uzh.ch/kelldata/trep-db/index.html ). For validation of the gene models, cDNA sequences coding the mature isoform of Rca were PCR amplified using attB sites flanking transcript specific primers (Supplementary table S3) with PrimeSTAR GXL polymerase (Clontech-Takara, USA) and cloned in pDONR201 vector using BP Clonase-II enzyme (Invitrogen, USA) as per the manufacturer’s recommendation. Multiple clones per construct were sequenced and the data was validated with the previously predicted gene models. Rca gene from banana, date palm, and pineapple was included in the analysis due to their position in the monocot evolution. Gene models were developed using the predicted transcripts for each of these genes from NCBI database (Table S1). To identify the Rca genes of wild rice species, AtRCA1 was used as a query sequence to search 11 Oryza species (Jacquemin et al., 2013 ) of Ensembl plants DNA database with default settings ( http://plants.ensembl.org/index.html ). For the Oryza Rca1 structural analysis, we did not include O. meridionalis , O. glaberrima , and O. longistaminata , because the sequences for the region was not complete in these three species. Plant growth condition and heat treatment Rice cultivars Nipponbare (GSOR 100) and N22 (PI 389102), maize inbred lines B73 (PI 550473) and Mo22 (PI 558534), and sorghum genotypes BTx623 (PI 564163), SC1019 (PI 656071), SC1047 (PI 656072) and B35 (PI 147224) were received from Germplasm Resources Information Network ( https://www.ars-grin.gov ). The wild rice species Oryza brachyantha (IRGC 101232) and Oryza australiensis (IRGC 101397) used in this study were received from Gerald E. Edwards, Washington State University, Pullman, WA, USA. The wheat cultivars Chinese Spring, Giza168, and PBW343 were used in the experiment. All plant materials were grown in Conviron PGR15 growth chamber equipped with high-intensity discharge lamps capable of Photosynthetically Active Radiation (PAR) of > 600 µmol/m 2 /sec before treating at high temperatures. Except for the rice and wild rice species, all plants were planted in 4 x 9-inch-deep pots using Sunshine LC1 potting mixture (SunGro Horticulture, Bellevue, WA, USA) supplemented with 14g Osmocote 14-14-14 slow releasing fertilizer. Cultivated and wild rice species were grown as mentioned in Giuliani et al. ( 2013 ). The wheat plants were grown under controlled temperature of 24/18°C with 16/8 hours of the day/night cycle. For rice maize, and sorghum genotypes the temperature was 25/22°C with 12/12 hours day/night cycle. Heat shock treatments were given to plants with fully expanded leaves (four-five weeks old). The chamber temperature was gradually increased (5°C/hour) from morning to reach the target temperatures and maintained for four hours before collecting leaf samples. To match the circadian rhythm, samples from control sets were also collected at the same time period. Leaf samples were frozen immediately in liquid nitrogen and stored in -80°C freezers for subsequent RNA and protein extractions. Immunoblot analysis Protein extraction and immunoblot analyses were performed as described in Yamori and von Caemmerer ( 2011 ) with some modifications. Protein concentrations were measured using Bradford reagent (Bio-Rad). For Fig. 4 , 2ul of the total protein was loaded per well on 1D-SDS-PAGE. Immunoblots were probed first with cotton anti-Rubisco activase (Agrisera, Catalog # AS10-700) antibody and later with horseradish peroxidase-conjugated secondary antibody as per the manufacturers’ recommendation. Immunological bindings were detected using Pierce ECL Western Blotting Substrate (Thermo Scientific) with Kodak X-ray film developing system. RNA extraction, cDNA preparation, and Real Time (RT) PCR analysis Total RNA from the normal and heat stress treated leaf samples were extracted using the hot phenol RNA extraction method. In brief, 100–200 mg frozen tissue samples were ground in liquid nitrogen and 0.75 ml of preheated (80°C) extraction buffer containing a mix of 100 mM Tris-HCl (pH 8.0), 100 mM Lithium Chloride (LiCl), 20 mM Ethylenediaminetetraacetic acid (EDTA), 1% Sodium dodecyl sulfate (SDS) prepared in Diethyl pyrocarbonate (DEPC) treated water and equal volume of phenol (pH 4.8) was added. Before centrifugation, the homogenates were vortexed, chloroform (0.35 ml) was added and vortexed till the mix turn whitish green. The resulted supernatant was separated, precipitated with 8M LiCl in ice for two hours and spun at 4°C for 30 min. The RNA pellets were washed with 2M LiCl and 70% ethanol and air dried. The pellets were dissolved in DEPC treated water and DNAse digestion was performed using Turbo™ DNAse kit (Thermo Fisher Scientific; Catalog # AM2238) as per the manufacturer’s recommendation. Purified RNA was loaded on a 1% agarose gel to check the quality and quantity of the sample. First strand cDNA was synthesized from the purified RNA samples using M-MLV Reverse transcriptase (Promega; Catalog # M1701) with anchored-oligo (dT) 18 primer as per the manufacturer’s recommendation. The cDNA samples were diluted 50 times and used in the RT-PCR reactions. All the RT-PCR analysis were performed using the KAPA SYBR® FAST qPCR master mix (Kapa Biosystems Inc; Catalog # KK4609) on LightCycler® 480 (Roche Diagnostics), as per the manufacturer’s instructions. The sequences of all the PCR primers used for the RT-PCR are given in Table S3. The relative expression for each sample was calculated using 2 − Δ C T , where Δ C T = ( C T gene of interest – C T internal control) (Schmittgen and Livak 2008) . Phylogenetic analysis Phylogenetic studies were performed using the putative RCA amino acid sequences retrieved from the annotated gene models. The complete list of the sequences used is listed in the SI. The analysis was carried out using the Maximum likelihood method in the Molecular Evolutionary Genetics Analysis version 7 (MEGA7) (Kumar et al., 2016 ). Alignment for tree construction was done using MUSCLE with the Gonnet scoring matrix. Reliability of the obtained trees was tested using 1000 bootstrap replicates. Declarations Conflict of interest statement The authors declare no conflict of interest. Author contributions RN and KSG designed the research, RN and KSK performed the experiments and data analysis and wrote the initial draft of manuscript. KSK, AM and KSG critically revised and prepared the final draft of the manuscript. Funding This work was supported by the USDA National Institute of Food and Agriculture, Hatch (project number WNP00449) and United States Agency for International Development Feed the Future Innovation Lab-Climate Resilient Wheat (Grant number AID-OAA-A-13-00008). Author contributions: RN and KSG designed the research, RN and KSK performed the experiments and data analysis and wrote the initial draft of manuscript. KSK, AM and KSG critically revised and prepared the final draft of the manuscript. Acknowledgements We sincerely thank Dr. G.N. Mohan Kumar, Department of Horticulture, Washington State University for providing advice on protein extraction and immunoblot analyses, Sheila Fitzgerald for proofreading the manuscript, and Dr. Gerald Edwards and Dr. Rita Giuliani, School of Biological Sciences, Washington State University for providing wild rice species and protocol for growing rice plants under control conditions. References Allakhverdiev SI, Kreslavski VD, Klimov V V., Los DA, Carpentier R, Mohanty P . 2008. Heat stress: An overview of molecular responses in photosynthesis. Photosynthesis Research 98 , 541–550. Ashraf M, Harris PJC . 2013. Photosynthesis under stressful environments: An overview. Photosynthetica 51 , 163–190. Beguiristain T, Grandbastien M-A, Puigdomènech P, Casacuberta JM . 2001. Three Tnt1 subfamilies show different stress-associated patterns of expression in tobacco. Consequences for retrotransposon control and evolution in plants. Plant Physiology 127 , 212–221. Carmo-silva AE, Salvucci ME . 2013. The regulatory properties of Rubisco activase differ among species and affect photosynthetic induction during light transitions. Plant physiology 161 , 1645–55. Carmo-Silva E, Scales JC, Madgwick PJ, Parry M a J . 2015. Optimizing Rubisco and its regulation for greater resource use efficiency. Plant, cell & environment 38 , 1817–32. Chase MW, Christenhusz MJM, Fay MF, et al. 2016. An update of the Angiosperm Phylogeny Group classification for the orders and families of flowering plants: APG IV. Botanical Journal of the Linnean Society 181 , 1–20. Crafts-Brandner SJ, Salvucci ME . 2000. Rubisco activase constrains the photosynthetic potential of leaves at high temperature and CO2. Proceedings of the National Academy of Sciences of the United States of America 97 , 13430–5. Crafts-Brandner SJ, Salvucci ME . 2002. Sensitivity of photosynthesis in a C4 plant, maize, to heat stress. Plant physiology 129 , 1773–1780. Dhaliwal AK, Mohan A, Gill KS . 2014. Comparative analysis of ABCB1 reveals novel structural and functional conservation between monocots and dicots. Frontiers in Plant Science 5 , 657. Feller U, Crafts-Brandner SJ, Salvucci ME . 1998. Moderately High Temperatures Inhibit Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase (Rubisco) Activase-Mediated Activation of Rubisco. Plant physiology 116 , 539–546. Feschotte C . 2008. Transposable elements and the evolution of regulatory networks. Nature Reviews Genetics 9 , 397–405. Ge S, Sang T, Lu BR, Hong DY . 1999. Phylogeny of rice genomes with emphasis on origins of allotetraploid species. Proceedings of the National Academy of Sciences of the United States of America 96 , 14400–14405. Giuliani R, Koteyeva N, Voznesenskaya E, Evans MA, Cousins AB, Edwards GE . 2013. Coordination of Leaf Photosynthesis, Transpiration, and Structural Traits in Rice and Wild Relatives (Genus Oryza). Plant physiology 162 , 1632–51. Guo Y-L, Ge S . 2005. Molecular phylogeny of Oryzeae ( Poaceae ) based on DNA sequences from chloroplast, mitochondrial, and nuclear genomes. American journal of botany 92 , 1548–58. Ito H, Gaubert H, Bucher E, Mirouze M, Vaillant I, Paszkowski J . 2011. An siRNA pathway prevents transgenerational retrotransposition in plants subjected to stress. Nature 472 , 115–119. Ito H, Yoshida T, Tsukahara S, Kawabe A . 2013. Evolution of the ONSEN retrotransposon family activated upon heat stress in Brassicaceae. Gene 518 , 256–261. Jacquemin J, Bhatia D, Singh K, Wing RA . 2013. The International Oryza Map Alignment Project: Development of a genus-wide comparative genomics platform to help solve the 9 billion-people question. Current Opinion in Plant Biology 16 , 147–156. Kellogg EA . 2001. Evolutionary history of the grasses. Plant physiology 125 , 1198–1205. Kobza J, Edwards GE . 1987. Influences of leaf temperature on photosynthetic carbon metabolism in wheat. Plant physiology 83 , 69–74. Kumar A, Li C, Portis A . 2009. Arabidopsis thaliana expressing a thermostable chimeric Rubisco activase exhibits enhanced growth and higher rates of photosynthesis at moderately high temperatures. Photosynthesis research 100 , 143–53. Kumar S, Stecher G, Tamura K . 2016. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular biology and evolution 33 , 1870–1874. Kurek I, Chang TK, Bertain SM, Madrigal A, Liu L, Lassner MW, Zhu G . 2007. Enhanced Thermostability of Arabidopsis Rubisco activase improves photosynthesis and growth rates under moderate heat stress. The Plant cell 19 , 3230–41. Law RD, Crafts-Brandner SJ . 1999. Inhibition and acclimation of photosynthesis to heat stress is closely correlated with activation of ribulose-1,5-bisphosphate Carboxylase/Oxygenase. Plant physiology 120 , 173–82. Law RD, Crafts-Brandner SJ . 2001. High temperature stress increases the expression of wheat leaf ribulose-1,5-bisphosphate carboxylase/oxygenase activase protein. Archives of biochemistry and biophysics 386 , 261–7. Lescot M, Déhais P, Thijs G, Marchal K, Moreau Y, Van de Peer Y, Rouzé P, Rombauts S . 2002. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic acids research 30 , 325–327. Lisch D . 2013. How important are transposons for plant evolution? Nature Reviews Genetics 14 , 49–61. Makarevitch I, Waters AJ, West PT, Stitzer M, Hirsch CN, Ross-Ibarra J, Springer NM . 2015. Transposable elements contribute to activation of maize genes in response to abiotic stress. PLoS Genetics 11 . Middleton CP, Senerchia N, Stein N, Akhunov ED, Keller B, Wicker T, Kilian B . 2014. Sequencing of chloroplast genomes from wheat, barley, rye and their relatives provides a detailed insight into the evolution of the triticeae tribe. PLoS ONE 9 . Morales A, Ortega-Delgado ML, Molina-Galan J, de Jimenez ES . 1999. Importance of Rubisco activase in maize productivity based on mass selection procedure. Journal of Experimental Botany 50 , 823–829. Nagai T, Makino A . 2009. Differences between rice and wheat in temperature responses of photosynthesis and plant growth. Plant & cell physiology 50 , 744–55. Nagarajan R, Gill KS . 2018. Evolution of Rubisco activase gene in plants. Plant Molecular Biology 96 , 69–87. Naito K, Zhang F, Tsukiyama T, Saito H, Hancock CN, Richardson AO, Okumoto Y, Tanisaka T, Wessler SR . 2009. Unexpected consequences of a sudden and massive transposon amplification on rice gene expression. Nature 461 , 1130–1134. Portis AR, Salvucci ME, Ogren WL . 1986. Activation of Ribulose bisphosphate Carboxylase/Oxygenase at physiological CO2 and Ribulosebisphosphate concentrations by Rubisco activase. Plant physiology 82 , 967–971. Ristic Z, Momcilovic I, Bukovnik U, Prasad PVV, Fu J, Deridder BP, Elthon TE, Mladenov N . 2009. Rubisco activase and wheat productivity under heat-stress conditions. Journal of experimental botany 60 , 4003–14. Rundle S, Zielinski R . 1991. Organization and expression of two tandemly oriented genes encoding ribulosebisphosphate carboxylase/oxygenase activase in barley. The Journal of biological chemistry 266 , 4677–4685. Salse J, Bolot S, Throude M, Jouffe V, Piegu B, Quraishi UM, Calcagno T, Cooke R, Delseny M, Feuillet C . 2008. Identification and Characterization of Shared Duplications between Rice and Wheat Provide New Insight into Grass Genome Evolution. The plant cell 20 , 11–24. Salvucci ME, van de Loo FJ, Stecher D . 2003. Two isoforms of Rubisco activase in cotton, the products of separate genes not alternative splicing. Planta 216 , 736–44. Salvucci ME, Portis AR, Ogren WL . 1985. A soluble chloroplast protein catalyzes ribulosebisphosphate carboxylase/oxygenase activation in vivo. Photosynthesis research 7 , 193–201. Scafaro AP, Bautsoens N, den Boer B, Van Rie J, Gallé A . 2019. A Conserved Sequence from Heat-Adapted Species Improves Rubisco Activase Thermostability in Wheat. Plant physiology 181 , 43–54. Scafaro AP, Gallé A, Van Rie J, Carmo-Silva E, Salvucci ME, Atwell BJ . 2016. Heat tolerance in a wild Oryza species is attributed to maintenance of Rubisco activation by a thermally stable Rubisco activase ortholog. New Phytologist 211 , 899–911. Scafaro AP, Haynes PA, Atwell BJ . 2010. Physiological and molecular changes in Oryza meridionalis Ng., a heat-tolerant species of wild rice. Journal of Experimental Botany 61 , 191–202. De Souza FSJ, Franchini LF, Rubinstein M . 2013. Exaptation of transposable elements into novel Cis-regulatory elements: Is the evidence always strong? Molecular Biology and Evolution 30 , 1239–1251. Swigoňová Z, Lai J, Ma J, Ramakrishna W, Llaca V, Bennetzen JL, Messing J . 2004. Close split of sorghum and maize genome progenitors. Genome Research 14 , 1916–1923. To KY, Suen DF, Chen SC . 1999. Molecular characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase activase in rice leaves. Planta 209 , 66–76. Vaughan DA, Morishima H, Kadowaki K . 2003. Diversity in the Oryza genus. Current Opinion in Plant Biology 6 , 139–146. Vogel JP, Garvin DF, Mockler TC, et al. 2010. Genome sequencing and analysis of the model grass Brachypodium distachyon. Nature 463 , 763–768. Wang D, Li X-F, Zhou Z-J, Feng X-P, Yang W-J, Jiang D-A . 2010. Two Rubisco activase isoforms may play different roles in photosynthetic heat acclimation in the rice plant. Physiologia plantarum 139 , 55–67. Werneke JM, Chatfield JM, Ogren WL . 1989. Alternative mRNA splicing generates the two ribulosebisphosphate carboxylase/oxygenase activase polypeptides in spinach and Arabidopsis. The Plant cell 1 , 815–825. Yamori W, von Caemmerer S . 2011. Quantification of Rubisco activase content in leaf extracts. Methods in molecular biology (Clifton, N.J.) 684 , 383–391. Yamori W, Suzuki K, Noguchi K, Nakai M, Terashima I . 2006. Effects of Rubisco kinetics and Rubisco activation state on the temperature dependence of the photosynthetic rate in spinach leaves from contrasting growth temperatures. Plant, Cell and Environment 29 , 1659–1670. Yin Z, Meng F, Song H, Wang X, Xu X, Yu D . 2010. Expression quantitative trait loci analysis of two genes encoding rubisco activase in soybean. Plant physiology 152 , 1625–37. Yin Z, Zhang Z, Deng D, Chao M, Gao Q, Wang Y, Yang Z, Bian Y, Hao D, Xu C . 2014. Characterization of Rubisco Activase Genes in Maize: An α-Isoform Gene Functions alongside a β-Isoform Gene. Plant physiology 164 , 2096–106. Zhang N, Kallis RP, Ewy RG, Portis AR . 2002. Light modulation of Rubisco in Arabidopsis requires a capacity for redox regulation of the larger Rubisco activase isoform. Proceedings of the National Academy of Sciences of the United States of America 99 , 3330–3334. Zhang Z, Komatsu S . 2000. Molecular cloning and characterization of cDNAs encoding two isoforms of ribulose-1,5-bisphosphate carboxylase/oxygenase activase in rice (Oryza sativa L.). Journal of Biochemistry 128 , 383–389. Zhang N, Portis AR . 1999. Mechanism of light regulation of Rubisco: a specific role for the larger Rubisco activase isoform involving reductive activation by thioredoxin-f. Proceedings of the National Academy of Sciences of the United States of America 96 , 9438–43. Zhang N, Schürmann P, Portis AR . 2001. Characterization of the regulatory function of the 46-kDa isoform of Rubisco activase from Arabidopsis. Photosynthesis Research 68 , 29–37. Zhao C, Liu B, Piao S, et al. 2017. Temperature increase reduces global yields of major crops in four independent estimates. Proceedings of the National Academy of Sciences 114 , 9326–9331. Tables Table 2. List of transposable elements identified in the grass Rca genic region Gene TE type Size (bp) Region E value No. of HSEs (sequence) ObRca1 Mutator-like (MULE) 336 Promoter 0 2 (aaaaaattta; caaatttttt) OsRca2 Mutator-like (MULE) 347 Promoter 0 1 (aaaaaatttg) OsRca2 MuDR-like (MITE) 320 Intron (5) 0 1 (gtaatttttt) TaRca1-A Gypsy (LTR) 319 Promoter 3e-43 0 TaRca1-B Mariner (MITE) 239 Promoter 1e-18 0 TaRca1 -D Copia (LTR) 967 Promoter 0 0 TaRca2 -D Mariner (MITE) 162 Promoter 5e-21 0 ZmRca1 Class II/III TE 368 Promoter 0 1 (agataattcg) SbRca1 CACTA-consensus 584 Promoter 2e-97 1 (acaaaatttc) Supplementary Files TableS1.docx TableS2.docx TableS3.docx Cite Share Download PDF Status: Published Journal Publication published 21 Dec, 2024 Read the published version in Plant Molecular Biology → Version 1 posted Editorial decision: Accept 28 Sep, 2024 Reviewers agreed at journal 10 Jul, 2024 Reviewers invited by journal 09 Jul, 2024 Editor invited by journal 06 Jul, 2024 Editor assigned by journal 04 Jul, 2024 First submitted to journal 03 Jul, 2024 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-4676428","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":324769419,"identity":"7e4ea26e-d867-4a9a-a7a1-10faacf071d5","order_by":0,"name":"Ragupathi Nagarajan","email":"","orcid":"","institution":"Washington State University","correspondingAuthor":false,"prefix":"","firstName":"Ragupathi","middleName":"","lastName":"Nagarajan","suffix":""},{"id":324769421,"identity":"331ee666-328f-407a-a3e0-858df3a87378","order_by":1,"name":"Kaviraj Singh Kahlon","email":"","orcid":"","institution":"Washington State University","correspondingAuthor":false,"prefix":"","firstName":"Kaviraj","middleName":"Singh","lastName":"Kahlon","suffix":""},{"id":324769423,"identity":"b5107c77-c634-4764-bf93-28aaf831a24e","order_by":2,"name":"Amita Mohan","email":"","orcid":"","institution":"Washington State University","correspondingAuthor":false,"prefix":"","firstName":"Amita","middleName":"","lastName":"Mohan","suffix":""},{"id":324769425,"identity":"38627e5a-a187-4e40-a134-a229a5394d66","order_by":3,"name":"Kulvinder S. Gill","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAtElEQVRIiWNgGAWjYBACCSA+8KDABsLjIVpLgkEaiVoYEgwOk6BFsv/4RaAt5xPnz0hgfPC2jQgt0hI5BUAttxM33EhgNpxLjBY5CZ4EkJbcDRIJbNK8RGnhPwPSci4X6DD230RpkWZIPwDUciC34UYCGzNRWiRn5IACObl+w5mHzZJzzhGhReL88ccfPlTYGcu3Jx/88KaMCC3AuDCAMhgbiFIPBOwPiFU5CkbBKBgFIxUAAGWAOkwTn7cHAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0001-7370-6184","institution":"Washington State University","correspondingAuthor":true,"prefix":"","firstName":"Kulvinder","middleName":"S.","lastName":"Gill","suffix":""}],"badges":[],"createdAt":"2024-07-02 21:17:17","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4676428/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4676428/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11103-024-01515-z","type":"published","date":"2024-12-21T15:57:25+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":61619666,"identity":"f1059882-065b-4de8-922f-84fa3cc68e6a","added_by":"auto","created_at":"2024-08-02 04:34:46","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":302359,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEvolution of Rubisco activase gene in cereals. \u003c/strong\u003eComparison of \u003cem\u003eRca \u003c/em\u003egene models from cereals showing structural differences, accumulation of heat shock elements and insertion various transposable elements with an evolutionarily predicted timeline. The \u003cem\u003eRca\u003c/em\u003e gene evolution model for grasses is adapted from Nagarajan and Gill (2017). The color bars on and below the gray dotted lines represent gene models and its corresponding alternate mRNA splicing models. Species with multiple gray lines represent RCA coding genes on different chromosomes. The a and bsymbols in the gene models denote the type of isoforms coded from the corresponding gene copy. In gene models, the thin black bar represents introns and thick light gray bars represent UTR regions. The green, yellow, brown, sky blue, red, navy blue and purple color bars represent exons from 1 to 7 of the ancestral \u003cem\u003eRca\u003c/em\u003egene, respectively. Bars with multiple colors represent fused exons that lost introns in comparison to the ancestral \u003cem\u003eRca\u003c/em\u003e gene model. The exons of non-functional rice \u003cem\u003eOsRca1 \u003c/em\u003eis shown as white bars. Inverted triangles in orange depict transposable element insertions and asterisk symbol (*) below the gray dotted lines represent heat shock elements in the promoter region. The asterisk symbol on the inverted orange triangles show the presence of heat shock elements in the transposable element sequences. The gene model at the top indicates the putative common grass ancestor \u003cem\u003eRca\u003c/em\u003e gene and predicted evolutionary changes based on \u003cem\u003eRca\u003c/em\u003egenes of different cereal crops. The evolutionary timescale on species divergence presented on the left side of gene models were based on previous estimates - “a” (Guo and Ge, 2005), “b” (Vogel \u003cem\u003eet al.\u003c/em\u003e, 2010), (Middleton \u003cem\u003eet al.\u003c/em\u003e, 2014), “c” (Swigoňová \u003cem\u003eet al.\u003c/em\u003e, 2004). The gene models were drawn to scale.\u003c/p\u003e","description":"","filename":"Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/3f1fb03176fd160070a77235.png"},{"id":61619293,"identity":"fd9c340a-fb04-4678-96f5-a217f01a88db","added_by":"auto","created_at":"2024-08-02 04:26:46","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":489714,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eStructure of Rubisco activase genes in different monocot species. a.\u003c/strong\u003e Taxonomical classification showing different monocot species (bold letters) of Commelinids clade used in this study. Angiosperm Phylogeny Group IV (APG-IV) system (Chase \u003cem\u003eet al.\u003c/em\u003e, 2016) was followed to show the classification. \u003cstrong\u003eb.\u003c/strong\u003e RCA coding gene models of banana (\u003cem\u003eMusa acuminata\u003c/em\u003e), date palm (\u003cem\u003ePhoenix dactylifera\u003c/em\u003e), and pineapple (\u003cem\u003eAnanas comosus\u003c/em\u003e) were drawn based on the comparative analysis of genomic and cDNA sequences (see materials and methods). The color bars on and below the gray dotted lines represent gene models and its corresponding alternate mRNA splicing models. The aand bin front of each gene model denote the type of isoform coded from that gene model. Species with two or more gray dotted lines represent RCA coding genes on different genomic locations or chromosomes. In gene models, the thin black bars represent introns and light gray bars represent UTR regions. The green, yellow, brown, sky blue, red, navy blue and purple color bars represent exons 1 to 7 respectively. The gene models were drawn to scale.\u003c/p\u003e","description":"","filename":"Fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/aef93b9f656e9dcf28b882df.png"},{"id":61619292,"identity":"ff80dd37-a6f4-47e8-b016-36242e778ad4","added_by":"auto","created_at":"2024-08-02 04:26:46","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":444218,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComparison of sequence deletions between the \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eRca1\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e gene of Oryza species. \u003c/strong\u003eComparative analysis showing nucleotide deletions and insertions in the \u003cem\u003eRca1\u003c/em\u003e gene of different Oryza species. In gene models, green, yellow, brown, sky blue, red, navy blue and purple color bars represent corresponding exonic regions from the ancestral \u003cem\u003eRca\u003c/em\u003e gene exon shown in Fig.1. Grey bars represent introns and their sizes (in basepairs) are mentioned below the double slash (//) of each gray bar. Upward black triangles above the gaps show nucleotide deletions and downward black triangles show insertions, corresponding to the \u003cem\u003eRca1\u003c/em\u003e sequence of \u003cem\u003eO.\u003c/em\u003e \u003cem\u003ebrachyantha\u003c/em\u003e. Red color triangles represent unique deletions or insertions identified in that particular species. The numbers above the triangles represent the nucleotide count on that particular addition or deletion (-) site. Box1 and Box2 represent unique deletions found in the box overlapping species.\u003c/p\u003e","description":"","filename":"Fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/576c006ed71fabe27e72b666.png"},{"id":61619301,"identity":"ec70b4d2-fef4-495e-b987-ba558045cdc7","added_by":"auto","created_at":"2024-08-02 04:26:46","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":364587,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eExpression pattern of various \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eRca\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e transcripts in cereals. \u003c/strong\u003eQuantitative Real-Time PCR (qRT-PCR) analysis showing the transcript levels of tandemly duplicated \u003cem\u003eRca\u003c/em\u003e genes of different genotypes of rice, wheat, maize and sorghum at normal (24/25°C), moderate (35/37°C) and severe heat stress (42/45°C) conditions. The \u003cem\u003eRca \u003c/em\u003etranscripts of rice, wild rice, maize, and sorghum were quantified using gene and alternate splicing specific primers. In case of wheat, the primers were specific to \u003cem\u003eRca1\u003c/em\u003e, \u003cem\u003eRca2\u003c/em\u003e and its alternatively spliced transcripts but not to the A, B and D homoeologous copies. The columns on the chart indicate the mean relative expression of \u003cem\u003eRca\u003c/em\u003e transcripts and the number on the x- axis represent different genotypes. Data represent the means and standard errors (SE) of three biological replicates (n=3). A complete list of primers is shown in Supplemental table S3. \u003cstrong\u003ea\u003c/strong\u003e. Effect of heat stress on the \u003cem\u003eRca\u003c/em\u003e genes of two rice cultivars Nipponbare and N22 (column #1 and 2) and two wild species \u003cem\u003eO. australiensis\u003c/em\u003e, and \u003cem\u003eO.\u003c/em\u003e \u003cem\u003ebrachyantha\u003c/em\u003e, (column #3 and 4). \u003cstrong\u003eb\u003c/strong\u003e. Response of wheat \u003cem\u003eTaRca1\u003c/em\u003e and \u003cem\u003eTaRca2 \u003c/em\u003eexpression to high temperature stress in cultivar Chinese Spring (column #1), Giza168 (column #2) and PBW343 (column #3). \u003cstrong\u003ec. \u003c/strong\u003eExpression pattern of \u003cem\u003eZmRca1\u003c/em\u003e and \u003cem\u003eZmRca1\u003c/em\u003e at normal and elevated temperatures in maize inbred lines B73 (column #1) and Mo22 (column #2). \u003cstrong\u003ed. \u003c/strong\u003eEffect of temperature stress on the expression of\u003cstrong\u003e \u003c/strong\u003e\u003cem\u003eSbRca1\u003c/em\u003e and \u003cem\u003eSbRca2 \u003c/em\u003eof B35 (column #1), BTx623 (column #2), SC1019 (column #3), and SC1047 (column #4).\u003c/p\u003e","description":"","filename":"Fig4.png","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/4244e2c2dfecd6938fd6e58b.png"},{"id":61619667,"identity":"3b6711ca-4067-4833-b27f-966e981059fe","added_by":"auto","created_at":"2024-08-02 04:34:46","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":584926,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eImmunoblot analysis showing expression pattern of cereal RCA isoforms at elevated temperatures. \u003c/strong\u003eProteins were extracted from control (25°C) and heat stressed (45°C) leaf samples of four weeks old rice (cv. Nipponbare), wheat (cv. Chinese spring), maize (B73 inbred) and sorghum (BTx623 inbred) lines. Plants grown under control conditions (25°C day/21°C night) were gradually exposed (5°C / 30 min) to heat stress and maintained at 45°C (45°C) for four hours. Leaf samples from control and heat-treated plants were collected at noon (six hours after the chamber lights were on) to avoid any effect of circadian rhythm on the gene expression. Samples were frozen immediately in liquid nitrogen and the total protein was extracted (see material and methods for details). An equal amount of total protein (2 mg) was loaded in 1D-SDS-PAGE for immunoblot analysis. Anti-RCA specific polyclonal antibody raised in rabbits (Agrisera, Catalog # AS10-700) was used as primary to detect RCA isoforms. The Coomassie Brilliant Blue R250 stained gel below the immunoblot shows the Rubisco large subunit.\u003c/p\u003e","description":"","filename":"Fig5.png","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/ea468bc1d60f49e3b922d7f1.png"},{"id":61619300,"identity":"a2887b28-cb88-4bbd-b8e6-a3b29317340f","added_by":"auto","created_at":"2024-08-02 04:26:46","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1259528,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eComparison of grass RCA sequences with wild and thermostable variants of Arabidopsis RCA. \u003c/strong\u003eMultiple sequence alignment showing a part of mature cereal RCA isoform (excluding chloroplast transit peptide) sequences with Arabidopsis wild type (AtRCA1) and thermostable RCA variant (AtRCA1-183H12). The amino acid substitution T274R, which provided thermostability of RCA in AtRCA1-183H12 variant, and its corresponding amino acid in cereal RCA isoforms are shown in the box. The exact Arginine (R) amino acid is marked in bold and underlined.\u003c/p\u003e","description":"","filename":"FIg6.png","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/4150a702841a9877410f8232.png"},{"id":61619295,"identity":"99e1680e-41a6-4b53-9caa-d990ac0380e6","added_by":"auto","created_at":"2024-08-02 04:26:46","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":273113,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMaximum Likelihood phylogenetic tree showing relationship between different monocot RCA proteins. \u003c/strong\u003eMonocot species used for the analysis are \u003cem\u003eAnanas comosus \u003c/em\u003e(Ac), \u003cem\u003ePhoenix dactylifera\u003c/em\u003e (Pd), \u003cem\u003eMusa acuminata\u003c/em\u003e (Ma), \u003cem\u003eTriticum aestivum\u003c/em\u003e (Ta), \u003cem\u003eHordeum vulgare\u003c/em\u003e (Hv), \u003cem\u003eBrachypodium distachyon\u003c/em\u003e (Bd), \u003cem\u003eOryza sativa\u003c/em\u003e (Os), \u003cem\u003eOryza brachyantha\u003c/em\u003e (Ob), \u003cem\u003eOryza puctata\u003c/em\u003e (Op), \u003cem\u003eSetaria italica\u003c/em\u003e(Si), \u003cem\u003eZea mays\u003c/em\u003e (Zm), and \u003cem\u003eSorghum bicolor\u003c/em\u003e (Sb). Scale bar represents the number of amino acid changes per site and the numbers at the nodes represent bootstrap values larger than 70%.\u003c/p\u003e","description":"","filename":"Fig7.png","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/0f3b86545a87b50a4430aa80.png"},{"id":72201744,"identity":"8c1ecc28-989c-4c11-ad6c-eefab89e786a","added_by":"auto","created_at":"2024-12-23 16:10:27","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":5286003,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/1fa0bb9a-54f0-4084-81fa-47b66424549b.pdf"},{"id":61619297,"identity":"50a5dbdb-572a-4288-8350-58361c21b8a7","added_by":"auto","created_at":"2024-08-02 04:26:46","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":14265,"visible":true,"origin":"","legend":"","description":"","filename":"TableS1.docx","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/381156e75d4a92947787433e.docx"},{"id":61619668,"identity":"3de79f26-5661-4020-aed4-0178f2869be8","added_by":"auto","created_at":"2024-08-02 04:34:46","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":39206,"visible":true,"origin":"","legend":"","description":"","filename":"TableS2.docx","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/e3ad15565f5a462454848645.docx"},{"id":61619299,"identity":"fbb0e5d3-87b5-4910-8a9d-b65f0c0af59c","added_by":"auto","created_at":"2024-08-02 04:26:46","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":15912,"visible":true,"origin":"","legend":"","description":"","filename":"TableS3.docx","url":"https://assets-eu.researchsquare.com/files/rs-4676428/v1/73eda10c629d7f6ddc85da8f.docx"}],"financialInterests":"","formattedTitle":"Tandemly Duplicated Rubisco Activase Genes of Cereals Show Differential Evolution and Response to Heat Stress","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe optimal temperature for photosynthesis varies among crops, from 25-30\u0026deg;C in wheat to 30-35\u0026deg;C in rice (Nagai and Makino, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Every degree-Celsius increase over the optimal temperature may result in up to 6% reduction in grain yield of wheat and 3.2% of rice (Zhao et al., \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Although many plant metabolic pathways are affected by heat stress, the effect on photosynthesis is critical as any constraint on the process will directly impact development, growth, biomass production, and crop yield. Plant photosynthetic machinery involves complex components and processes, many of which including Rubisco activation, enzymes and proteins of photosystems, membrane permeability and ATP generation are vulnerable to high temperature (Allakhverdiev et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). The critical temperature and its duration affecting each photosynthetic process often differ among species (Feller et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1998\u003c/span\u003e). Plants exposed to temperatures above 40\u0026deg;C for extended periods show irreversible damage to the permeability of thylakoid membranes and electron transport chain, resulting in photosynthesis inhibition. The photosystems and electron transport are usually not affected until 40\u0026deg;C, whereas for many species including some important crop plants including wheat the Rubisco activation level and photosynthetic rate (Pn) start to decline even at moderately elevated temperatures (Feller et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1998\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe functional state of Rubisco is maintained by Rubisco activase (RCA) that removes inhibitory sugar phosphates from the active sites of Rubisco with energy derived from ATP hydrolysis (Salvucci et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e1985\u003c/span\u003e). RCA is a nuclear encoded, cytosol synthesized chloroplast protein, generally consisted of two immunologically related polypeptides called α isoform (45 to 46 kDa) and β isoform (41 to 43 kDa). The size difference between the two isoforms is due to the presence of a redox regulated C-terminal extension (CTE) domain that is present in the α isoform but is lacking in the β form. In most plants, both α and β isoforms of RCA are capable of activating Rubisco (Werneke et al., \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1989\u003c/span\u003e; Rundle and Zielinski, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; To et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Salvucci et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). Shown in Arabidopsis, the two cysteine (Cys) residues located in the CTE domain of RCA α isoform make a disulfide bond under oxidizing conditions. This significantly increases its sensitivity to physiological level of ADP/ATP ratio and affects both ATP hydrolysis and Rubisco activation (Zhang and Portis, \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Zhang et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). The major function of larger isoform is to regulate the activity of smaller RCA isoforms under low light/dark conditions in order to conserve energy. The equimolar concentrations of the larger and the smaller isoforms, completely inhibited the activity of smaller isoform under higher ADP/ATP ratio in light conditions (Zhang et al., \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). Interestingly, tobacco, which expresses only the β isoform, is also sensitive to physiological levels of ADP/ATP ratio and responds similar to species with both isoforms (Carmo-Silva and Salvucci, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Selection for genotypes with higher \u003cem\u003eRca\u003c/em\u003e expression levels at normal conditions in soybean, maize, and some other crops showed a positive correlation with Rubisco activity, photosynthetic rate, and seed yield (Morales et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Yin et al., \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2010\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn-vitro analysis has shown that while the RUBISCO enzyme is stable even at 50\u003csup\u003eo\u003c/sup\u003eC, RCA is very sensitive to heat stress and its activity start to decline at temperatures over 35\u003csup\u003eo\u003c/sup\u003eC (Crafts-Brandner and Salvucci, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). As a result, at higher temperatures RCA is unable to keep pace with faster Rubisco deactivation thus constrains photosynthesis (Crafts-Brandner and Salvucci, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). In crops including cotton and wheat, even a moderate level of heat stress decreases the Rubisco activation levels, which correlated with reduced net photosynthesis, increased RuBP level, and decreased levels of 3-phosphoglycerate in leaves (Law and Crafts-Brandner, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1999\u003c/span\u003e). Independent studies on various crops have shown that photosynthetic heat acclimation is directly associated with the acclimation of heat sensitive RCA to high temperature (Law and Crafts-Brandner, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Yamori et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). This conclusion was further supported by the studies showing increase in the thermal tolerance of Arabidopsis photosynthesis by improving RCA thermostability (Kurek et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Kumar et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePreviously, we showed that the \u003cem\u003eRca\u003c/em\u003e gene structure is highly conserved in higher plants, and the common grass ancestor \u003cem\u003eRca\u003c/em\u003e, similar to Arabidopsis \u003cem\u003eAtRca1\u003c/em\u003e, contained seven exons and six introns, enabling expression of the RCA α isoform with redox regulated CTE domain coded from the seventh exon (Nagarajan and Gill, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The alternatively spliced mRNA of the same gene contains a premature stop codon and produces a shorter redox insensitive RCA β isoform (Werneke et al., \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1989\u003c/span\u003e). We also reported that the \u003cem\u003eRca\u003c/em\u003e gene is tandemly duplicated (\u003cem\u003eRca1\u003c/em\u003e and \u003cem\u003eRca2\u003c/em\u003e) among grass species and the duplication event probably occurred in the common grass ancestor, but before the divergence of subfamilies (Nagarajan and Gill, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Heat stress induced changes in the RCA expression pattern at protein level were reported earlier in rice (Scafaro et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), wheat (Feller et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Law and Crafts-Brandner, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Ristic et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), and maize (Crafts-Brandner and Salvucci, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Ristic et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). High temperature stress altered the ratio of α and β isoform levels in rice (Wang et al., \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), but induced new RCA isoform expression in wheat and maize (Law and Crafts-Brandner, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Crafts-Brandner and Salvucci, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Ristic et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). A recent study in wheat showed that heat stress altered the expression pattern of \u003cem\u003eRca\u003c/em\u003e genes (Scafaro et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Later, (Degen \u003cem\u003eet al.\u003c/em\u003e, 2020) showed a single amino acid substitution (M159I) in TaRCA2-β isoform could increase the thermal stability of the enzymatic activity by up to 5\u0026deg;C. However, the heat stress induced expression pattern of RCA isoforms in important cereal crops and key evolutionary changes causing the differential gene expression are still not well studied. Earlier, we proposed an evolutionary model for tandemly duplicated \u003cem\u003eRca\u003c/em\u003e genes of the grass species (Nagarajan and Gill, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). We also showed that after the duplication event, one of the gene copies, \u003cem\u003eRca1\u003c/em\u003e, have lost the third, the fifth and the sixth introns, which also abolished the alternative splicing mechanism of the \u003cem\u003eRca\u003c/em\u003e transcripts. Other changes observed in Rca1 among various grass species included nonfunctional \u003cem\u003eRca1\u003c/em\u003e in some Oryzoideae species; 2) loss of \u003cem\u003eRca1\u003c/em\u003e sequences coding the redox regulated CTE domain in Pooideae including that in Brachypodium, barley, and wheat and the loss of various introns (Ragarajan and Gill, 2018). 3) loss of \u003cem\u003eRca2\u003c/em\u003e sequences coding for the redox regulated CTE domain in Panicoideae species (maize, sorghum and foxtail millet), and 4) loss of various introns in both \u003cem\u003eRca1\u003c/em\u003e and \u003cem\u003eRca2\u003c/em\u003e of these crops. Whereas, the second gene copy, \u003cem\u003eRca2\u003c/em\u003e, did not amass any major changes in its gene structure although loss of some introns and sequences coding for the redox regulated CTE domain in Panicoideae species including maize, sorghum and foxtail millet was observed.\u003c/p\u003e \u003cp\u003eIn the present study, first, we studied the origin of the tandem duplication of \u003cem\u003eRca\u003c/em\u003e genes. Specifically, we tracked the evolution of the rice \u003cem\u003eOsRca1\u003c/em\u003e by comparing the sequences of various Oryzoideae species. Second, we studied the effect of heat stress on the expression pattern of tandemly duplicated cereal \u003cem\u003eRca\u003c/em\u003e gene copies both at the mRNA as well as at the protein level and investigated possible causes (i.e. promoter elements) of the variation. Finally, we performed a phylogenetic study to analyze the rate of amino acid substitution and the evolutionary relationship of different monocot RCA isoforms. The information was then used to understand the role of tandemly duplicated \u003cem\u003eRca\u003c/em\u003e genes in Rubisco activation, and photosynthetic acclimation to heat stress in important cereal crops.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e \u003cb\u003eTandem duplication of\u003c/b\u003e \u003cb\u003eRca\u003c/b\u003e \u003cb\u003egene occurred after the divergence of Poales families\u003c/b\u003e\u003c/p\u003e \u003cp\u003eWith the objective to accurately determine origin of the tandem duplication in the RCA gene, we evaluated the \u003cem\u003eRca\u003c/em\u003e gene structure in three distantly related monocot species: banana (\u003cem\u003eMusa acuminata\u003c/em\u003e), date palm (\u003cem\u003ePhoenix dactylifera\u003c/em\u003e) and pineapple (\u003cem\u003eAnanus comosus;\u003c/em\u003e family: Bromeliaceae; order Poales). Unlike the grass family, no tandem duplication of the \u003cem\u003eRca\u003c/em\u003e gene was observed in any of these species. Instead, these species contained three \u003cem\u003eRca\u003c/em\u003e gene copies on separate chromosomes (table S1). The \u003cem\u003eRca\u003c/em\u003e gene copies in banana were located on chromosomes 2, 10 and 11, whereas in date palm, the copies were present on two different unplaced scaffolds NW_008246537.1 (1.6 Mbp) and NW_008246711.1 (0.506 Mbp) and were not tandemly duplicated as was seen in grasses. The gene copies in pineapple were located in the linkage groups (LG) LG14, LG18, and LG23. Otherwise, the structure of the gene copies was similar to those of other higher plants. In all three species, the exon sizes and relative order were highly conserved among the three copies as was seen in other plant species. Major variation was however observed for intron length (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb). Based on these observations, we concluded that the gene duplication event occurred after the divergence of the Poales families and is probably specific to the Poaceae.\u003c/p\u003e \u003cp\u003e \u003cb\u003eRca\u003c/b\u003e \u003cb\u003egene structure among cereals\u003c/b\u003e\u003c/p\u003e \u003cp\u003eAfter evaluating Rca gene structure among various grass family members, we identified and sequence confirmed \u003cem\u003eRca\u003c/em\u003e gene copies from seven (\u003cem\u003eO. brachyantha\u003c/em\u003e, \u003cem\u003eO. australiensis\u003c/em\u003e, \u003cem\u003eO. sativa\u003c/em\u003e, \u003cem\u003eT. aestivum\u003c/em\u003e, \u003cem\u003eZ. mays\u003c/em\u003e, \u003cem\u003eS. bicolor\u003c/em\u003e, and \u003cem\u003eSetaria italica\u003c/em\u003e) species. Since rice showed major variation for the \u003cem\u003eRca\u003c/em\u003e gene structure, we also obtained sequence of seven cultivated rice accessions and two wild rice species \u003cem\u003e(Oryza australiensis\u003c/em\u003e and \u003cem\u003eOryza brachyantha\u003c/em\u003e). For each accession, both genomic and cDNA copies were cloned and sequenced. While the \u003cem\u003eRca2\u003c/em\u003e sequence from one of the accessions was reported earlier (To et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Scafaro et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), we cloned and sequenced the \u003cem\u003eRca1\u003c/em\u003e gene from \u003cem\u003eO. australiensis\u003c/em\u003e (GenBank Acc. MH115971), and \u003cem\u003eO. brachyantha\u003c/em\u003e (GenBank Acc. MH220418), and \u003cem\u003eRca2\u003c/em\u003e gene from \u003cem\u003eO. brachyantha\u003c/em\u003e (GenBank Acc. MH660917).\u003c/p\u003e \u003cp\u003eThe structural comparison of \u003cem\u003eRca1\u003c/em\u003e gene copy across various cultivated and wild rice species revealed presence of large deletions in the exons 2, 3, 4, 5, 6 and 7 which appeared to have caused frameshift mutation in the \u003cem\u003eRca1\u003c/em\u003e of cultivated rice species (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). In other species, between 11 to 13 independent deletions were detected in all four exons, resulting in 157 to a total loss of 218 nucleotides. These \u003cem\u003eRca1\u003c/em\u003e deletions appear to have occurred multiple times, but in different time periods during the evolution. For example, \u003cem\u003eO. barthi\u003c/em\u003e and \u003cem\u003eO. glumipatula\u003c/em\u003e have a 15 bp deletion on the third exon, whereas \u003cem\u003eO. nivara\u003c/em\u003e, \u003cem\u003eO. sativa\u003c/em\u003e, and \u003cem\u003eO. rufipogon\u003c/em\u003e showed a 39 bp deletion (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e; Box1). Similarly, the fourth exon of \u003cem\u003eRca1\u003c/em\u003e of \u003cem\u003eO. nivara\u003c/em\u003e, \u003cem\u003eO. sativa\u003c/em\u003e, and \u003cem\u003eO. rufipogon\u003c/em\u003e contains two additional deletions of 13 and 24 nucleotides (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e; Box 2), which were absent in other wild species. Additionally, species specific indels in the \u003cem\u003eRca1\u003c/em\u003e gene were also identified. The six bp addition in the second exon of \u003cem\u003eO. punctata Rca1\u003c/em\u003e, a 25 bp deletion in the fourth exon of \u003cem\u003eO. nivara\u003c/em\u003e, and a nine bp instead of the 14 bp deletion in the second exon of \u003cem\u003eO. glumipatula Rca1\u003c/em\u003e occurred probably after the divergence of the species. However, it is not clear how the 24 bp deletion is absent in \u003cem\u003eO. sativa\u003c/em\u003e ssp. \u003cem\u003eindica\u003c/em\u003e but present in most other species including \u003cem\u003eO. sativa\u003c/em\u003e ssp. \u003cem\u003ejaponica\u003c/em\u003e. Interestingly, the intron lengths of \u003cem\u003eRca1\u003c/em\u003e were determined before the major \u003cem\u003eOryza\u003c/em\u003e species divergence and maintained thereafter (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Alignment of RCA2 amino acid sequences of these species showed 100% similarity, except for \u003cem\u003eO. brachyantha\u003c/em\u003e and \u003cem\u003eO. punctata\u003c/em\u003e, which had only 95.5% and 97.4% similarities with the other \u003cem\u003eOryza\u003c/em\u003e species respectively.\u003c/p\u003e \u003cp\u003e \u003cb\u003eRca1\u003c/b\u003e \u003cb\u003egene structure in Oryzoideae subfamily\u003c/b\u003e\u003c/p\u003e \u003cp\u003eFrom the comparative analysis of \u003cem\u003eRca\u003c/em\u003e genes in Poaceae, it was evident that both Pooideae and Panicoideae subfamilies have two functional \u003cem\u003eRca\u003c/em\u003e gene copies present in tandem (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The \u003cem\u003eOsRca1\u003c/em\u003e copy contained multiple small deletions that seem to have occurred after the divergence of these sub-families. Comparison of the two wild and seven cultivated rice species revealed the presence of a putative \u003cem\u003eRca1\u003c/em\u003e gene in all these species. Structurally, the \u003cem\u003eRca1\u003c/em\u003e gene in \u003cem\u003eO. australiensis\u003c/em\u003e, \u003cem\u003eO. brachyantha\u003c/em\u003e, and \u003cem\u003eO. punctata\u003c/em\u003e contained four exons with slight size variation for the three introns (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The remaining five species including the two accessions of cultivated rice showed length of the three introns were remarkably conserved. The exons of these five species showed 11 to 13 deletions (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Nine of these deletions were common to all five species and the remaining were specific to one or more species. At the predicted protein level, the largest protein among these three species was predicted to be 466aa in \u003cem\u003eO. punctata\u003c/em\u003e. The predicted protein in \u003cem\u003eO. australiensis\u003c/em\u003e is 465aa as compared to 458 in \u003cem\u003eO. brachyantha\u003c/em\u003e. Unlike the RCA1 of Pooideae members, these predicted peptides also contain the light sensitive CTE domain as identified in the RCA1 of the Panicoideae members.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eThermostability conferring amino acids in RCA1 isoform of cereals\u003c/h2\u003e \u003cp\u003ePrevious studies showed that \u003cem\u003eArabidopsis\u003c/em\u003e transgenic plants expressing RCA variants with certain amino acid substitutions were heat tolerant, and the modified Arabidopsis RCA showed up to 10\u0026deg;C increase in stability and Rubisco activation activity (Kurek et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). One such substitution T274R (Threonine 274 Arginine) in the variant 183H12, which alone could provide the maximal heat stability and activity, was identified in all three copies of TaRCA1 and ZmRCA2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). Other natural variants observed in the same position were glutamine (Q) in SbRCA1, SbRCA2, and ZmRCA1, and lysine (K) in ObRCA1 and ObRCA2 of \u003cem\u003eO. brachyantha\u003c/em\u003e, and in all of the RCA2 isoforms of rice and wheat. Based on these results, we hypothesize that the isoform expressed during heat stress may have better thermal stability than the isoforms expressed at normal conditions because of the putative thermal tolerant amino acid substitutions. The role of these natural RCA variants in thermal tolerance is not known, and further experiments are needed to identify the significance of these sequence changes in providing thermal tolerance.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003ePhylogenetic analysis of monocot RCA\u003c/h2\u003e \u003cp\u003ePhylogenetic analysis performed with the alignment of RCA protein from distantly related monocots, using the MEGA7 maximum likelihood method, revealed different clusters of RCAs, based on the sequence similarity and evolutionary pattern (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e). In deep nodes, the phylogenetic relationship is unclear, and the bootstrap (BS) values are low (\u0026lt;\u0026thinsp;70%). The comparative analysis showed how the duplicated \u003cem\u003eRca\u003c/em\u003e genes evolved in cereals. Thus, we did not collapse the deep nodes with lower statistical support. Based on the phylogenetic analysis, RCA isoforms of banana, date palm and pineapple, and the cereal isoforms, formed distinct clades, with cereals further comprising of RCA1 and RCA2 from the tandemly duplicated genes. Although the RCA isoforms of banana (MaRCA1, 2 and 3), date palm (PdRCA1 and 2), and pineapple (AcRCA1, 2 and 3) were clustered together, the subgroup pattern within the cluster suggests that the genes coding these isoforms were duplicated or triplicated after the divergence of monocot orders. Similarly, the RCA1 and RCA2 of cereals formed different clades, but within each clade, the isoforms of closely related species, or species from a shared family, showed less divergence with high BS values. The branch and individual leaf lengths show the rate of substitution, the RCA1 isoforms of wild rice and Panicoideae species showed a higher rate of changes than did the RCA1 of Pooideae. Together, these results suggest that after the divergence for a common monocot ancestor, the \u003cem\u003eRca\u003c/em\u003e gene copy number increased either through polyploidization or tandem duplication and evolved at different rates in the studied species.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eExpression pattern of RCA isoforms in cereals\u003c/h2\u003e \u003cp\u003eDetailed analysis of 302 ESTs from banana (\u003cem\u003eMusa acuminata\u003c/em\u003e) and 331 from pineapple (\u003cem\u003eAnanus comosus\u003c/em\u003e) showed that in both banana and pineapple, only one of the three copies codes for RCA-α, whereas the other two copies code for only the β isoform (Supplementary Table S1). EST sequences showed no evidence of alternative splicing in the RCA-α coding copy, suggesting a possible loss of the splice junction during evolution. In comparison, alternate splicing was observed in date palm (\u003cem\u003ePhoenix dactylifera)\u003c/em\u003e based on the analysis of 305 ESTs. One of the two \u003cem\u003eRca\u003c/em\u003e copies, \u003cem\u003ePdRca1\u003c/em\u003e, codes for both RCA-α and RCA-β isoforms via alternative splicing, whereas the other gene copy (\u003cem\u003ePdRca2\u003c/em\u003e) codes only for the RCA-α isoform.\u003c/p\u003e \u003cp\u003eEvaluating the EST data, \u003cem\u003eMaRca1-α, MaRca2-β\u003c/em\u003e, and \u003cem\u003eMaRca3-β\u003c/em\u003e of Banana has 101, 101 and 100 ESTs respectively; \u003cem\u003ePdRca1-α, PdRca1-β and PdRca2-α\u003c/em\u003e of date palm has 103, 102 and 100 ESTs respectively and \u003cem\u003eAcRca1-α, AcRca2-β and AcRca3-β\u003c/em\u003e of pineapple has 110, 115 and 106 ESTs respectively. Based on this data, the three splice variants appear to be equally expressed in these three plants. With the objective to compare expression of the gene(s) at the RNA level with that at the protein level, and to identify the relative contribution of the two tandemly duplicated \u003cem\u003eRca\u003c/em\u003e gene copies, we performed gene specific expression analyses using quantitative Real Time PCR (qRT-PCR) for both \u003cem\u003eRca1\u003c/em\u003e and \u003cem\u003eRca2\u003c/em\u003e genes of \u003cem\u003eO. brachyantha\u003c/em\u003e, \u003cem\u003eO. australiensis\u003c/em\u003e, \u003cem\u003eO. sativa\u003c/em\u003e, \u003cem\u003eT. aestivum\u003c/em\u003e, \u003cem\u003eZ. mays\u003c/em\u003e, and \u003cem\u003eS. bicolor\u003c/em\u003e at normal as well as at high temperature conditions (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). In cultivated rice (\u003cem\u003eO. sativa\u003c/em\u003e), either under heat stress or normal plant growth conditions, no expression was observed for the \u003cem\u003eRca1\u003c/em\u003e copy in either of the two cultivars that were used for the analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ea). \u003cem\u003eO. australiensis\u003c/em\u003e showed some expression of \u003cem\u003eRca1\u003c/em\u003e under normal growth conditions and the expression reduced both under 35\u003csup\u003eo\u003c/sup\u003eC and 45\u003csup\u003eo\u003c/sup\u003eC treatment. \u003cem\u003eO. brachyantha\u003c/em\u003e on the other hand showed a different expression pattern for \u003cem\u003eRca1\u003c/em\u003e where its expression was very low under normal plant growth conditions but increased under the heat stress conditions. \u003cem\u003eRca1\u003c/em\u003e expression was however significantly less than that for \u003cem\u003eRca2\u003c/em\u003e. The \u003cem\u003eRca2\u003c/em\u003e copy in cultivated rice showed very high expression level under normal plant growth conditions and its expression reduced dramatically under heat stress conditions. There was significant difference between the two cultivated rice lines for the expression of \u003cem\u003eRca2\u003c/em\u003e copy. The two rice wild relatives showed dramatically higher expression for \u003cem\u003eRca2\u003c/em\u003e under heat stress. Even between the two wild relatives, \u003cem\u003eO. brachyantha\u003c/em\u003e showed significantly higher expression at 45\u003csup\u003eo\u003c/sup\u003eC treatment as compared to \u003cem\u003eO. australiensis\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eHeat stress also affected the alternative splicing mechanism, as the reduction in the levels of transcripts coding RCA2α varied during heat stress (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). In rice, \u003cem\u003eOsRca2\u003c/em\u003e gene expresses \u003cem\u003eOsRca2α\u003c/em\u003e and \u003cem\u003eOsRca2β\u003c/em\u003e transcripts via alternate splicing mechanism. At normal conditions, the level of \u003cem\u003eOsRca2β\u003c/em\u003e transcripts was many hundred-folds higher than that of the \u003cem\u003eOsRca2α\u003c/em\u003e transcripts, but at 42\u0026deg;C, the \u003cem\u003eOsRca2α\u003c/em\u003e transcript level was either equal or higher than that of the \u003cem\u003eOsRca2β\u003c/em\u003e transcripts. Interestingly, \u003cem\u003eObRca2α\u003c/em\u003e transcripts were stable at all studied temperatures, however, \u003cem\u003eObRca2β\u003c/em\u003e showed more than two-fold reduction at higher temperatures. These results suggest that the temperature increase, at least among various rice species, favors transcripts coding RCA2α isoform. Even though the transcripts encoding OsRCA2α and OsRCA2β showed a significant reduction at 45\u0026deg;C in rice, the quantity of both the isoforms from the heat stressed leaves is comparable or higher to their control leaves (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe expression pattern of the \u003cem\u003eRca\u003c/em\u003e gene copies in wheat was very different from that observed in rice. \u003cem\u003eRca1\u003c/em\u003e showed essentially no expression at normal plant growth conditions, but its expression increased dramatically under heat stress conditions (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eb). The expression pattern was very similar among the three cultivars (Chinese Spring, Giza168 and PBW343) although significant differences were observed among cultivars. In all three cultivars, transcript level for \u003cem\u003eRca2\u003c/em\u003e copy was significant at all three temperature conditions although its level at 37\u003csup\u003eo\u003c/sup\u003eC was significantly higher than either of the other two conditions. Genotypic differences were significant among the three genotypes at each temperature condition. Between the two isoforms of \u003cem\u003eRca2\u003c/em\u003e, transcript level of \u003cem\u003eRca2α\u003c/em\u003e isoform was much lower than the \u003cem\u003eRca2β\u003c/em\u003e form and stayed about the same over the three temperature treatments (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eb). The \u003cem\u003eRca2β\u003c/em\u003e form showed significant increase at 37\u003csup\u003eo\u003c/sup\u003eC treatment but reduction at 42\u003csup\u003eo\u003c/sup\u003eC.\u003c/p\u003e \u003cp\u003eBoth in maize and sorghum, \u003cem\u003eRca1\u003c/em\u003e copy made only the alpha isoform of the gene whereas the \u003cem\u003eRca2\u003c/em\u003e copy made the beta form (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ec and d). In both of these species, the beta form was made at all three temperatures although significant differences were observed both among temperatures as well as among genotypes. The alpha form showed essentially no accumulation at 25\u003csup\u003eo\u003c/sup\u003eC, slight increase at 35\u003csup\u003eo\u003c/sup\u003eC treatment and dramatic increase at 45\u003csup\u003eo\u003c/sup\u003eC. In general, the genotypic differences for relative expression of the two isoforms were much higher than that observed among the wheat genotypes.\u003c/p\u003e \u003cp\u003eUsing the same plant samples as that used for real-time expression analysis, RCA protein level was also studied using immunoblot analysis with an RCA specific antibody (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Except for wheat, all of the studied cereals expressed a 42 kDa RCAβ isoform at normal as well as under heat stress conditions whereas the 46 kDa RCAα isoform was expressed only during the heat stress conditions (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). In comparison to Rubisco, the expression level of wheat RCAβ was significantly higher at the normal temperature as compared to that under heat stress. Relative to RCAβ, expression of the RCAα is much lower at normal conditions but is undetectable after the heat treatment. Although it is difficult to accurately quantify, expression levels of both Rubisco, as well as RCA of rice, maize, and sorghum, increased after 45\u0026deg;C treatment as compared to that at the normal temperature. Wheat, on the other hand did not show the 46 kDa RCAα isoform under heat stress conditions along with a significant reduction in the RCAβ level. With the current analysis, it was not possible to determine if the 42 kDa RCAβ isoform of wheat is expressed from \u003cem\u003eTaRca1\u003c/em\u003e or the \u003cem\u003eTaRca2\u003c/em\u003e copy.\u003c/p\u003e \u003cp\u003e \u003cb\u003eRegulatory elements in the\u003c/b\u003e \u003cb\u003eRca\u003c/b\u003e \u003cb\u003egenes of cereals\u003c/b\u003e\u003c/p\u003e \u003cp\u003eAs the two \u003cem\u003eRca\u003c/em\u003e gene copies of cereals showed very different expression pattern, we looked for differences in expression control elements between promoters of the two copies. We evaluated regulatory elements in ~\u0026thinsp;1.5 kb region upstream of the transcription start site of the two \u003cem\u003eRca\u003c/em\u003e gene copies of the studies species along with the intronic sequences, with a focus on heat regulated elements (material and methods). Along with the core promoter elements, this analysis showed various putative cis-elements including that for circadian rhythm, light, hormone and biotic and abiotic stress responsives (Table S2). The \u003cem\u003eRca1\u003c/em\u003e copy showed insertion of heat shock elements (HSE) in its promoter with wild rice (\u003cem\u003eO. brachyantha\u003c/em\u003e) having three HSEs. These HSEs were missing in the \u003cem\u003eRca1\u003c/em\u003e promoter of cultivated rice (\u003cem\u003eO. sativa\u003c/em\u003e). Interestingly, \u003cem\u003eRca2\u003c/em\u003e of \u003cem\u003eO. sativa\u003c/em\u003e have HSEs in its promoter and in the 5th intron. Transposable element (TE) of size 366bp, 347bp and 320 bp were present in the promoter of \u003cem\u003eRca1\u003c/em\u003e, promoter of \u003cem\u003eRca2\u003c/em\u003e and in the 5th intron of \u003cem\u003eRca2\u003c/em\u003e, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). In wheat, \u003cem\u003eRca1\u003c/em\u003e promoter of the \u003cem\u003eA\u003c/em\u003e and the \u003cem\u003eB\u003c/em\u003e genome showed one HSEs and the \u003cem\u003eD\u003c/em\u003e genome has two. There were no HSEs present in the promoter of \u003cem\u003eRca2\u003c/em\u003e of wheat but promoter of D genome showed insertion of 162 bp TE. In maize (\u003cem\u003eZ. mays\u003c/em\u003e) and sorghum (\u003cem\u003eS. bicolor\u003c/em\u003e), both \u003cem\u003eRca1\u003c/em\u003e and \u003cem\u003eRca2\u003c/em\u003e had insertion of HSEs with sorghum having maximum of 5 HSEs in the promoter of \u003cem\u003eRca1\u003c/em\u003e. Promoter of \u003cem\u003eRca2\u003c/em\u003e had two HSEs in sorghum and one in maize. TE were also present in the promoter of \u003cem\u003eRca1\u003c/em\u003e in both maize and sorghum (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e)\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eAdaptation of organisms to the changing environmental conditions via modifications in the basic biological processes is a common feature of evolution. For better adaptation, an organism often relies on new gene mutations which may develop neo-functions, however, that could also cause detrimental effects if the mutated gene is essential for the organism\u0026rsquo;s survival. Gene and genome duplications provide the base genetic material for the origin of novel genes or trait development during evolution.\u003c/p\u003e\n\u003cp\u003eTandem duplication of \u003cem\u003eRca\u0026nbsp;\u003c/em\u003egenes was first reported in barley\u0026nbsp;(Rundle and Zielinski, 1991)\u0026nbsp;and later in wheat\u0026nbsp;(Carmo-Silva \u003cem\u003eet al.\u003c/em\u003e, 2015). In rice, DNA gel blot analysis with cDNA coding RCA as a probe showed the presence of more than one \u003cem\u003eRca\u003c/em\u003e gene in the genome\u0026nbsp;(Zhang and Komatsu, 2000). We\u0026nbsp;(Nagarajan and Gill, 2018)\u0026nbsp;showed that the tandem duplication event occurred in the common grass ancestor and the structural changes then accumulated after the divergence of the Poaceae families. In this study we have shown that this tandem duplication of \u003cem\u003eRca\u003c/em\u003e gene is Poaceae-specific and probably occurred after the divergence of Poales order as one (Bromeliaceae) of the 16 families did not have the duplication. It would be possible to pinpoint the exact timeline of the duplication event once the genome sequence information for the other 14 families is available.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003ePhylogenetic and fossil data analyses have suggested that the grass sub-families probably diverged from a common ancestor between 50-70 million years ago (MYA)\u0026nbsp;(Kellogg, 2001; Salse \u003cem\u003eet al.\u003c/em\u003e, 2008). The tandem duplication event of \u003cem\u003eRca\u003c/em\u003e genes might have occurred before the divergence of the grass sub-families as the Oryzoideae, Pooideae, and Panicoideae members have the duplicated genes (Fig. 2). The conserved heat induced expression pattern of \u003cem\u003eRca1\u003c/em\u003e in all three sub-families suggests that the promoter elements required for heat responsive expression might have integrated into the \u003cem\u003eRca1\u003c/em\u003e promoter of the grass ancestor. Later, species specific TE insertions added additional HSEs in the promoter region and further altered the gene expression. Effect of TE insertions on the neighboring gene expression is a well-documented phenomenon in plants and other organisms\u0026nbsp;(see reviews, Feschotte, 2008; Lisch, 2013; De Souza et al., 2013). Similarly, integration of biotic and abiotic responsive cis-regulatory elements in the promoter regions of genes by TE was also reported in plants\u0026nbsp;(Beguiristain \u003cem\u003eet al.\u003c/em\u003e, 2001; Naito \u003cem\u003eet al.\u003c/em\u003e, 2009), including HSEs\u0026nbsp;(Ito \u003cem\u003eet al.\u003c/em\u003e, 2013). The insertions of TEs with HSE elements in duplicated \u003cem\u003eRca\u003c/em\u003e gene regions of cereals could be the source for evolutionary adaptation through altered gene expression. Heat stress induced changes in the RCA expression pattern were reported earlier in rice\u0026nbsp;(Scafaro \u003cem\u003eet al.\u003c/em\u003e, 2010; Wang \u003cem\u003eet al.\u003c/em\u003e, 2010), wheat\u0026nbsp;(Feller \u003cem\u003eet al.\u003c/em\u003e, 1998; Law and Crafts-Brandner, 2001; Ristic \u003cem\u003eet al.\u003c/em\u003e, 2009), and maize\u0026nbsp;(Crafts-Brandner and Salvucci, 2002; Ristic \u003cem\u003eet al.\u003c/em\u003e, 2009). Our copy specific \u003cem\u003eRca\u003c/em\u003e transcript quantification along with immunoblot analysis have specifically revealed expression pattern of each of the \u003cem\u003eRca\u003c/em\u003e copies and of its splice varients. We have also shown differences and similarlities among \u003cem\u003eRca\u003c/em\u003e of various species for a differential response to heat stress. In comparison to maize and sorghum, the wheat \u003cem\u003eRca\u003c/em\u003e showed almost an opposite accumulation pattern of different \u003cem\u003eRca\u003c/em\u003e isoforms and splice varients (Figures 4\u0026amp;5). \u0026nbsp;Role of these differences in explaining differences in heat stress tolerance among wheat, maize and sorghum needs to be validated.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eOne of the important findings of this study is the identification of a functional \u003cem\u003eRca1\u003c/em\u003e gene copy among wild rice species. The genus \u003cem\u003eOryza\u003c/em\u003e is composed of 23 recognized species, including two cultivated (\u003cem\u003eO. sativa\u003c/em\u003e and \u003cem\u003eO. glaberrima\u003c/em\u003e) and 21 wild rice species with 10 different genomes (AA, BB, CC, BBCC, CCDD, EE, FF, GG, HHJJ, and HHKK)\u0026nbsp;(Ge \u003cem\u003eet al.\u003c/em\u003e, 1999; Vaughan \u003cem\u003eet al.\u003c/em\u003e, 2003). Our analysis on the non-functionalization of \u003cem\u003eRca1\u0026nbsp;\u003c/em\u003eshowed that the species \u003cem\u003eO. sativa\u003c/em\u003e, \u003cem\u003eO. glaberrima\u003c/em\u003e, \u003cem\u003eO. barthii\u003c/em\u003e, \u003cem\u003eO. glumipatula\u003c/em\u003e, \u003cem\u003eO. nivara\u003c/em\u003e, and \u003cem\u003eO. rufipogon\u003c/em\u003e with AA genome have multiple deletions in the exonic sequences thus have non-functional \u003cem\u003eRca1\u003c/em\u003e. Interestingly, \u003cem\u003eO. punctata\u0026nbsp;\u003c/em\u003ewith BB genome, \u003cem\u003eO. australiensis\u003c/em\u003e with EE, and \u003cem\u003eO. brachyantha\u003c/em\u003e with FF genome contain a complete and functional \u003cem\u003eRca1\u003c/em\u003e gene. These results suggest that the non-functionalization of \u003cem\u003eRca1\u003c/em\u003e in the AA genome species occurred after the divergence of \u003cem\u003eOryza\u003c/em\u003e species and also specifically in the AA genome lineage. Recent research on thermal tolerance of wild rice species \u003cem\u003eO. australiensis\u0026nbsp;\u003c/em\u003eshowed unaffected net photosynthetic and leaf elongation rates, even at 45\u0026deg;C, whereas \u003cem\u003eO. meridionalis\u0026nbsp;\u003c/em\u003eand cultivated rice \u003cem\u003eO. sativa\u003c/em\u003e showed intermediate tolerance to \u0026gt; 50% decline in net photosynthesis and leaf growth rates\u0026nbsp;(Scafaro \u003cem\u003eet al.\u003c/em\u003e, 2016). Rubisco activation levels of these species at high temperature correlated with the net photosynthesis and leaf elongation rate. Interestingly, the thermal stability of recombinant OaRCA2 was similar to the OsRCA2 of cultivated rice in the \u003cem\u003ein vitro\u0026nbsp;\u003c/em\u003eassays, but the leaf extracted RCA of \u003cem\u003eO. australiensis\u003c/em\u003e showed no Rubisco activation inhibition at temperatures as high as 42\u0026deg;C, whereas rice RCA inhibition occurred at temperatures above 34\u0026deg;C. The authors characterized the \u003cem\u003eOaRca2\u003c/em\u003e of \u003cem\u003eO. australiensis\u003c/em\u003e and compared its thermal stability with that of rice \u003cem\u003eOsRca2\u003c/em\u003e. They speculated that some unknown factor(s) in the leaf extract could contribute to its higher thermal activity and stability\u0026nbsp;(Scafaro \u003cem\u003eet al.\u003c/em\u003e, 2016). Based on our \u003cem\u003eRca1\u003c/em\u003e comparative analysis of \u003cem\u003eOryza\u003c/em\u003e sp. and heat stress induced expression of \u003cem\u003eOaRca1\u003c/em\u003e of \u003cem\u003eO. australiensis\u003c/em\u003e,\u003cem\u003e\u0026nbsp;\u003c/em\u003ewe\u003cem\u003e\u0026nbsp;\u003c/em\u003epropose that the presence of OaRCA1 could be one of the reasons for increased thermal stability of Rubisco activation and unaffected net photosynthetic observed in the above-mentioned study. However, further experiments are needed to characterize the \u003cem\u003eOaRca1\u003c/em\u003e and utilize this gene in rice breeding through alien introgression to improve photosynthetic thermal tolerance of rice cultivars.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eVarious heat shock elements (HSEs) present in the sequences upstream of the transcription start sites (TSS) were identified in \u003cem\u003eObRca1\u0026nbsp;\u003c/em\u003eof \u003cem\u003eO. brachyantha\u003c/em\u003e, \u003cem\u003eOsRca2\u0026nbsp;\u003c/em\u003eof rice, all three copies of wheat \u003cem\u003eTaRca1\u003c/em\u003e, both \u003cem\u003eZmRca1\u003c/em\u003e and \u003cem\u003eZmRca2\u003c/em\u003e of maize, and both \u003cem\u003eSbRca1\u003c/em\u003e and \u003cem\u003eSbRca2\u003c/em\u003e of sorghum (Fig. 1). The number of HSEs also varied among species with the promoter of \u003cem\u003eSbRca1\u0026nbsp;\u003c/em\u003econtaining up to five, \u003cem\u003eObRca1\u003c/em\u003e containing three, \u003cem\u003eTaRca1-D\u003c/em\u003e and \u003cem\u003eSbRca2\u0026nbsp;\u003c/em\u003econtaining two, and the remaining ones showing only a single HSE in the upstream region. It is tempting to speculate a correlation of higher level of heat stress tolerance in sorghum and wild rice with higher number of HSE\u0026rsquo;s but additional experimentation is needed to confirm.\u003c/p\u003e\n\u003cp\u003ePrevious studies have shown that certain transposable elements are preferentially inserted into the regulatory sequences of various genes and play an important role in the expression control of the neighboring genes\u0026nbsp;(Makarevitch \u003cem\u003eet al.\u003c/em\u003e, 2015). In some cases, heat stress induced gene expression was acquired from TE insertion events\u0026nbsp;(Ito \u003cem\u003eet al.\u003c/em\u003e, 2011). The duplicated \u003cem\u003eRca\u003c/em\u003e genes of closely related yet evolutionarily diverged cereals, and their heat stress induced expression pattern may serve as an ideal model for understanding different TE insertions and their role in the stress regulated expression. To identify TEs in the genic region, we used the duplicated gene sequences, along with 2Kb sequence flanking on both sides, as the query to search the crop specific TE database. We identified independent TE insertion events in the promoter sequence of all six \u003cem\u003eRca1\u003c/em\u003e copies of the studied species (Table 2). Similarly, there were TE insertions in the promoter and the fifth intron of rice \u003cem\u003eOsRca2\u003c/em\u003e, and in the promoter of \u003cem\u003eTaRca2-D\u0026nbsp;\u003c/em\u003ecopy (Fig. 1). Interestingly, many of these TEs contain HSEs; these TEs inserted in different time periods of evolution served as a potential source for the HSEs and may have caused heat stress induced expression pattern in the \u003cem\u003eRca1\u003c/em\u003e genes. A detailed list of the identified TEs, their sizes, and locations relative to the genes is given in Table 2.\u003c/p\u003e\n\u003cp\u003eIn conclusion, the comparative and expression analyses of RCA coding genes of important cereal crops revealed\u0026nbsp;a tandem gene duplication, which occurred in the common grass ancestor subsequently led to neo-functionalization and evolved to respond differently during heat stress. One of the two copies of the tandem duplication maintained a very high level of conservation whereas the second copy showed tremendous divergence to evolve species specific function of the gene. We identified the insertion of various transposable elements harboring heat responsive elements in the promoter region. These results suggest that one of the duplicated \u003cem\u003eRca\u003c/em\u003e genes showing divergence, which could be responsible for the heat induced expression pattern and thermal stability of these heat tolerant cereal crops. Understanding these evolutionary changes are important as this knowledge can be used to engineer future plants with better and wider adaptability.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eGene model prediction and validation\u003c/h2\u003e \u003cp\u003eThe genomic regions comprising \u003cem\u003eRca\u003c/em\u003e genes of banana, date palm, and pineapple were searched using the well characterized 474 aa sequence of AtRCA1 (AT2G39730.1) (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.arabidopsis.org/\u003c/span\u003e\u003cspan address=\"https://www.arabidopsis.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) as the query in Phytozome 11 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://phytozome.jgi.doe.gov\u003c/span\u003e\u003cspan address=\"https://phytozome.jgi.doe.gov\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and NCBI database. We selected banana, date palm, and pineapple because of its position in the monocot evolution with sequenced genome at the time of analysis. Gene models were developed using the predicted transcripts for each of these genes from NCBI database (Table S1). The \u003cem\u003eRca\u003c/em\u003e gene models for rice (cv. Nipponbare), wheat (cv. Chinese spring), maize (inbred B73), and sorghum (inbred BTx623) were adopted from our previous study (Nagarajan and Gill, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). To identify the \u003cem\u003eRca\u003c/em\u003e genes of wild rice species, AtRCA1 was used as a query sequence to search 11 \u003cem\u003eOryza\u003c/em\u003e species (Jacquemin et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) of Ensembl plants DNA database with default settings (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://plants.ensembl.org/index.html\u003c/span\u003e\u003cspan address=\"http://plants.ensembl.org/index.html\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). For the \u003cem\u003eOryza Rca1\u003c/em\u003e structural analysis, we did not include \u003cem\u003eO. meridionalis\u003c/em\u003e, \u003cem\u003eO. glaberrima\u003c/em\u003e, and \u003cem\u003eO. longistaminata\u003c/em\u003e, because the sequences for the region was not complete in these three species. The criteria used for searching true orthologs in this study were described elsewhere (Dhaliwal et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). For identification of cis-regulatory elements in the promoter region, we used the 1.5 Kb upstream sequence from ATG start codon for each gene to search in PlantCARE database (Lescot et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). The transposable elements for \u003cem\u003eOryza\u003c/em\u003e species were identified from RiTE-DB (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.genome.arizona.edu/cgi-bin/rite/index.cgi\u003c/span\u003e\u003cspan address=\"http://www.genome.arizona.edu/cgi-bin/rite/index.cgi\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e); wheat from cereal repeat database (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://wheat.pw.usda.gov/ggpages/Repeats/\u003c/span\u003e\u003cspan address=\"http://wheat.pw.usda.gov/ggpages/Repeats/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), maize from MaizeGDB (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.maizegdb.org\u003c/span\u003e\u003cspan address=\"http://www.maizegdb.org\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), and sorghum from TREP database (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://botserv2.uzh.ch/kelldata/trep-db/index.html\u003c/span\u003e\u003cspan address=\"http://botserv2.uzh.ch/kelldata/trep-db/index.html\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). For validation of the gene models, cDNA sequences coding the mature isoform of Rca were PCR amplified using \u003cem\u003eattB\u003c/em\u003e sites flanking transcript specific primers (Supplementary table S3) with PrimeSTAR GXL polymerase (Clontech-Takara, USA) and cloned in pDONR201 vector using BP Clonase-II enzyme (Invitrogen, USA) as per the manufacturer\u0026rsquo;s recommendation. Multiple clones per construct were sequenced and the data was validated with the previously predicted gene models. \u003cem\u003eRca\u003c/em\u003e gene from banana, date palm, and pineapple was included in the analysis due to their position in the monocot evolution. Gene models were developed using the predicted transcripts for each of these genes from NCBI database (Table S1).\u003c/p\u003e \u003cp\u003eTo identify the \u003cem\u003eRca\u003c/em\u003e genes of wild rice species, AtRCA1 was used as a query sequence to search 11 \u003cem\u003eOryza\u003c/em\u003e species (Jacquemin et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) of Ensembl plants DNA database with default settings (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://plants.ensembl.org/index.html\u003c/span\u003e\u003cspan address=\"http://plants.ensembl.org/index.html\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). For the \u003cem\u003eOryza Rca1\u003c/em\u003e structural analysis, we did not include \u003cem\u003eO. meridionalis\u003c/em\u003e, \u003cem\u003eO. glaberrima\u003c/em\u003e, and \u003cem\u003eO. longistaminata\u003c/em\u003e, because the sequences for the region was not complete in these three species.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003ePlant growth condition and heat treatment\u003c/h2\u003e \u003cp\u003eRice cultivars Nipponbare (GSOR 100) and N22 (PI 389102), maize inbred lines B73 (PI 550473) and Mo22 (PI 558534), and sorghum genotypes BTx623 (PI 564163), SC1019 (PI 656071), SC1047 (PI 656072) and B35 (PI 147224) were received from Germplasm Resources Information Network (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ars-grin.gov\u003c/span\u003e\u003cspan address=\"https://www.ars-grin.gov\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). The wild rice species \u003cem\u003eOryza brachyantha\u003c/em\u003e (IRGC 101232) and \u003cem\u003eOryza australiensis\u003c/em\u003e (IRGC 101397) used in this study were received from Gerald E. Edwards, Washington State University, Pullman, WA, USA. The wheat cultivars Chinese Spring, Giza168, and PBW343 were used in the experiment. All plant materials were grown in Conviron PGR15 growth chamber equipped with high-intensity discharge lamps capable of Photosynthetically Active Radiation (PAR) of \u0026gt;\u0026thinsp;600 \u0026micro;mol/m\u003csup\u003e2\u003c/sup\u003e/sec before treating at high temperatures. Except for the rice and wild rice species, all plants were planted in 4 x 9-inch-deep pots using Sunshine LC1 potting mixture (SunGro Horticulture, Bellevue, WA, USA) supplemented with 14g Osmocote 14-14-14 slow releasing fertilizer. Cultivated and wild rice species were grown as mentioned in Giuliani et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The wheat plants were grown under controlled temperature of 24/18\u0026deg;C with 16/8 hours of the day/night cycle. For rice maize, and sorghum genotypes the temperature was 25/22\u0026deg;C with 12/12 hours day/night cycle. Heat shock treatments were given to plants with fully expanded leaves (four-five weeks old). The chamber temperature was gradually increased (5\u0026deg;C/hour) from morning to reach the target temperatures and maintained for four hours before collecting leaf samples. To match the circadian rhythm, samples from control sets were also collected at the same time period. Leaf samples were frozen immediately in liquid nitrogen and stored in -80\u0026deg;C freezers for subsequent RNA and protein extractions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eImmunoblot analysis\u003c/h2\u003e \u003cp\u003eProtein extraction and immunoblot analyses were performed as described in Yamori and von Caemmerer (\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) with some modifications. Protein concentrations were measured using Bradford reagent (Bio-Rad). For Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e, 2ul of the total protein was loaded per well on 1D-SDS-PAGE. Immunoblots were probed first with cotton anti-Rubisco activase (Agrisera, Catalog # AS10-700) antibody and later with horseradish peroxidase-conjugated secondary antibody as per the manufacturers\u0026rsquo; recommendation. Immunological bindings were detected using Pierce ECL Western Blotting Substrate (Thermo Scientific) with Kodak X-ray film developing system.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eRNA extraction, cDNA preparation, and Real Time (RT) PCR analysis\u003c/h2\u003e \u003cp\u003eTotal RNA from the normal and heat stress treated leaf samples were extracted using the hot phenol RNA extraction method. In brief, 100\u0026ndash;200 mg frozen tissue samples were ground in liquid nitrogen and 0.75 ml of preheated (80\u0026deg;C) extraction buffer containing a mix of 100 mM Tris-HCl (pH 8.0), 100 mM Lithium Chloride (LiCl), 20 mM Ethylenediaminetetraacetic acid (EDTA), 1% Sodium dodecyl sulfate (SDS) prepared in Diethyl pyrocarbonate (DEPC) treated water and equal volume of phenol (pH 4.8) was added. Before centrifugation, the homogenates were vortexed, chloroform (0.35 ml) was added and vortexed till the mix turn whitish green. The resulted supernatant was separated, precipitated with 8M LiCl in ice for two hours and spun at 4\u0026deg;C for 30 min. The RNA pellets were washed with 2M LiCl and 70% ethanol and air dried. The pellets were dissolved in DEPC treated water and DNAse digestion was performed using Turbo\u0026trade; DNAse kit (Thermo Fisher Scientific; Catalog # AM2238) as per the manufacturer\u0026rsquo;s recommendation. Purified RNA was loaded on a 1% agarose gel to check the quality and quantity of the sample. First strand cDNA was synthesized from the purified RNA samples using M-MLV Reverse transcriptase (Promega; Catalog # M1701) with anchored-oligo (dT)\u003csub\u003e18\u003c/sub\u003e primer as per the manufacturer\u0026rsquo;s recommendation. The cDNA samples were diluted 50 times and used in the RT-PCR reactions. All the RT-PCR analysis were performed using the KAPA SYBR\u0026reg; FAST qPCR master mix (Kapa Biosystems Inc; Catalog # KK4609) on LightCycler\u0026reg; 480 (Roche Diagnostics), as per the manufacturer\u0026rsquo;s instructions. The sequences of all the PCR primers used for the RT-PCR are given in Table S3. The relative expression for each sample was calculated using 2\u003csup\u003e\u0026minus;\u0026thinsp;Δ\u003cem\u003eC\u003c/em\u003eT\u003c/sup\u003e, where Δ\u003cem\u003eC\u003c/em\u003e\u003csub\u003eT\u003c/sub\u003e = (\u003cem\u003eC\u003c/em\u003e\u003csub\u003eT\u003c/sub\u003e gene of interest \u0026ndash; \u003cem\u003eC\u003c/em\u003e\u003csub\u003eT\u003c/sub\u003e internal control) (Schmittgen and Livak 2008) .\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003ePhylogenetic analysis\u003c/h2\u003e \u003cp\u003ePhylogenetic studies were performed using the putative RCA amino acid sequences retrieved from the annotated gene models. The complete list of the sequences used is listed in the SI. The analysis was carried out using the Maximum likelihood method in the Molecular Evolutionary Genetics Analysis version 7 (MEGA7) (Kumar et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Alignment for tree construction was done using MUSCLE with the Gonnet scoring matrix. Reliability of the obtained trees was tested using 1000 bootstrap replicates.\u003c/p\u003e \u003c/div\u003e "},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eConflict of interest statement\u003c/h2\u003e \u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e \u003ch2\u003eAuthor contributions\u003c/h2\u003e \u003cp\u003eRN and KSG designed the research, RN and KSK performed the experiments and data analysis and wrote the initial draft of manuscript. KSK, AM and KSG critically revised and prepared the final draft of the manuscript.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis work was supported by the USDA National Institute of Food and Agriculture, Hatch (project number WNP00449) and United States Agency for International Development Feed the Future Innovation Lab-Climate Resilient Wheat (Grant number AID-OAA-A-13-00008).\u003c/p\u003e\u003ch2\u003eAuthor contributions:\u003c/h2\u003e \u003cp\u003eRN and KSG designed the research, RN and KSK performed the experiments and data analysis and wrote the initial draft of manuscript. KSK, AM and KSG critically revised and prepared the final draft of the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e \u003cp\u003eWe sincerely thank Dr. G.N. Mohan Kumar, Department of Horticulture, Washington State University for providing advice on protein extraction and immunoblot analyses, Sheila Fitzgerald for proofreading the manuscript, and Dr. Gerald Edwards and Dr. Rita Giuliani, School of Biological Sciences, Washington State University for providing wild rice species and protocol for growing rice plants under control conditions.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003e\u003cstrong\u003eAllakhverdiev SI, Kreslavski VD, Klimov V V., Los DA, Carpentier R, Mohanty P\u003c/strong\u003e. 2008. Heat stress: An overview of molecular responses in photosynthesis. Photosynthesis Research \u003cstrong\u003e98\u003c/strong\u003e, 541\u0026ndash;550.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eAshraf M, Harris PJC\u003c/strong\u003e. 2013. Photosynthesis under stressful environments: An overview. Photosynthetica \u003cstrong\u003e51\u003c/strong\u003e, 163\u0026ndash;190.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eBeguiristain T, Grandbastien M-A, Puigdom\u0026egrave;nech P, Casacuberta JM\u003c/strong\u003e. 2001. Three Tnt1 subfamilies show different stress-associated patterns of expression in tobacco. Consequences for retrotransposon control and evolution in plants. Plant Physiology \u003cstrong\u003e127\u003c/strong\u003e, 212\u0026ndash;221.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eCarmo-silva AE, Salvucci ME\u003c/strong\u003e. 2013. The regulatory properties of Rubisco activase differ among species and affect photosynthetic induction during light transitions. Plant physiology \u003cstrong\u003e161\u003c/strong\u003e, 1645\u0026ndash;55.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eCarmo-Silva E, Scales JC, Madgwick PJ, Parry M a J\u003c/strong\u003e. 2015. Optimizing Rubisco and its regulation for greater resource use efficiency. Plant, cell \u0026amp; environment \u003cstrong\u003e38\u003c/strong\u003e, 1817\u0026ndash;32.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eChase MW, Christenhusz MJM, Fay MF, \u003cem\u003eet al.\u003c/em\u003e\u003c/strong\u003e 2016. An update of the Angiosperm Phylogeny Group classification for the orders and families of flowering plants: APG IV. Botanical Journal of the Linnean Society \u003cstrong\u003e181\u003c/strong\u003e, 1\u0026ndash;20.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eCrafts-Brandner SJ, Salvucci ME\u003c/strong\u003e. 2000. Rubisco activase constrains the photosynthetic potential of leaves at high temperature and CO2. Proceedings of the National Academy of Sciences of the United States of America \u003cstrong\u003e97\u003c/strong\u003e, 13430\u0026ndash;5.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eCrafts-Brandner SJ, Salvucci ME\u003c/strong\u003e. 2002. Sensitivity of photosynthesis in a C4 plant, maize, to heat stress. Plant physiology \u003cstrong\u003e129\u003c/strong\u003e, 1773\u0026ndash;1780.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eDhaliwal AK, Mohan A, Gill KS\u003c/strong\u003e. 2014. Comparative analysis of ABCB1 reveals novel structural and functional conservation between monocots and dicots. Frontiers in Plant Science \u003cstrong\u003e5\u003c/strong\u003e, 657.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eFeller U, Crafts-Brandner SJ, Salvucci ME\u003c/strong\u003e. 1998. Moderately High Temperatures Inhibit Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase (Rubisco) Activase-Mediated Activation of Rubisco. Plant physiology \u003cstrong\u003e116\u003c/strong\u003e, 539\u0026ndash;546.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eFeschotte C\u003c/strong\u003e. 2008. Transposable elements and the evolution of regulatory networks. Nature Reviews Genetics \u003cstrong\u003e9\u003c/strong\u003e, 397\u0026ndash;405.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eGe S, Sang T, Lu BR, Hong DY\u003c/strong\u003e. 1999. Phylogeny of rice genomes with emphasis on origins of allotetraploid species. Proceedings of the National Academy of Sciences of the United States of America \u003cstrong\u003e96\u003c/strong\u003e, 14400\u0026ndash;14405.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eGiuliani R, Koteyeva N, Voznesenskaya E, Evans MA, Cousins AB, Edwards GE\u003c/strong\u003e. 2013. Coordination of Leaf Photosynthesis, Transpiration, and Structural Traits in Rice and Wild Relatives (Genus Oryza). Plant physiology \u003cstrong\u003e162\u003c/strong\u003e, 1632\u0026ndash;51.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eGuo Y-L, Ge S\u003c/strong\u003e. 2005. Molecular phylogeny of \u003cem\u003eOryzeae\u003c/em\u003e (\u003cem\u003ePoaceae\u003c/em\u003e) based on DNA sequences from chloroplast, mitochondrial, and nuclear genomes. American journal of botany \u003cstrong\u003e92\u003c/strong\u003e, 1548\u0026ndash;58.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eIto H, Gaubert H, Bucher E, Mirouze M, Vaillant I, Paszkowski J\u003c/strong\u003e. 2011. An siRNA pathway prevents transgenerational retrotransposition in plants subjected to stress. Nature \u003cstrong\u003e472\u003c/strong\u003e, 115\u0026ndash;119.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eIto H, Yoshida T, Tsukahara S, Kawabe A\u003c/strong\u003e. 2013. Evolution of the ONSEN retrotransposon family activated upon heat stress in Brassicaceae. Gene \u003cstrong\u003e518\u003c/strong\u003e, 256\u0026ndash;261.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eJacquemin J, Bhatia D, Singh K, Wing RA\u003c/strong\u003e. 2013. The International Oryza Map Alignment Project: Development of a genus-wide comparative genomics platform to help solve the 9 billion-people question. Current Opinion in Plant Biology \u003cstrong\u003e16\u003c/strong\u003e, 147\u0026ndash;156.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eKellogg EA\u003c/strong\u003e. 2001. Evolutionary history of the grasses. Plant physiology \u003cstrong\u003e125\u003c/strong\u003e, 1198\u0026ndash;1205.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eKobza J, Edwards GE\u003c/strong\u003e. 1987. Influences of leaf temperature on photosynthetic carbon metabolism in wheat. Plant physiology \u003cstrong\u003e83\u003c/strong\u003e, 69\u0026ndash;74.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eKumar A, Li C, Portis A\u003c/strong\u003e. 2009. Arabidopsis thaliana expressing a thermostable chimeric Rubisco activase exhibits enhanced growth and higher rates of photosynthesis at moderately high temperatures. Photosynthesis research \u003cstrong\u003e100\u003c/strong\u003e, 143\u0026ndash;53.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eKumar S, Stecher G, Tamura K\u003c/strong\u003e. 2016. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular biology and evolution \u003cstrong\u003e33\u003c/strong\u003e, 1870\u0026ndash;1874.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eKurek I, Chang TK, Bertain SM, Madrigal A, Liu L, Lassner MW, Zhu G\u003c/strong\u003e. 2007. Enhanced Thermostability of Arabidopsis Rubisco activase improves photosynthesis and growth rates under moderate heat stress. The Plant cell \u003cstrong\u003e19\u003c/strong\u003e, 3230\u0026ndash;41.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eLaw RD, Crafts-Brandner SJ\u003c/strong\u003e. 1999. Inhibition and acclimation of photosynthesis to heat stress is closely correlated with activation of ribulose-1,5-bisphosphate Carboxylase/Oxygenase. Plant physiology \u003cstrong\u003e120\u003c/strong\u003e, 173\u0026ndash;82.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eLaw RD, Crafts-Brandner SJ\u003c/strong\u003e. 2001. High temperature stress increases the expression of wheat leaf ribulose-1,5-bisphosphate carboxylase/oxygenase activase protein. Archives of biochemistry and biophysics \u003cstrong\u003e386\u003c/strong\u003e, 261\u0026ndash;7.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eLescot M, D\u0026eacute;hais P, Thijs G, Marchal K, Moreau Y, Van de Peer Y, Rouz\u0026eacute; P, Rombauts S\u003c/strong\u003e. 2002. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic acids research \u003cstrong\u003e30\u003c/strong\u003e, 325\u0026ndash;327.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eLisch D\u003c/strong\u003e. 2013. How important are transposons for plant evolution? Nature Reviews Genetics \u003cstrong\u003e14\u003c/strong\u003e, 49\u0026ndash;61.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eMakarevitch I, Waters AJ, West PT, Stitzer M, Hirsch CN, Ross-Ibarra J, Springer NM\u003c/strong\u003e. 2015. Transposable elements contribute to activation of maize genes in response to abiotic stress. PLoS Genetics \u003cstrong\u003e11\u003c/strong\u003e.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eMiddleton CP, Senerchia N, Stein N, Akhunov ED, Keller B, Wicker T, Kilian B\u003c/strong\u003e. 2014. Sequencing of chloroplast genomes from wheat, barley, rye and their relatives provides a detailed insight into the evolution of the triticeae tribe. PLoS ONE \u003cstrong\u003e9\u003c/strong\u003e.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eMorales A, Ortega-Delgado ML, Molina-Galan J, de Jimenez ES\u003c/strong\u003e. 1999. Importance of Rubisco activase in maize productivity based on mass selection procedure. Journal of Experimental Botany \u003cstrong\u003e50\u003c/strong\u003e, 823\u0026ndash;829.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eNagai T, Makino A\u003c/strong\u003e. 2009. Differences between rice and wheat in temperature responses of photosynthesis and plant growth. Plant \u0026amp; cell physiology \u003cstrong\u003e50\u003c/strong\u003e, 744\u0026ndash;55.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eNagarajan R, Gill KS\u003c/strong\u003e. 2018. Evolution of Rubisco activase gene in plants. Plant Molecular Biology \u003cstrong\u003e96\u003c/strong\u003e, 69\u0026ndash;87.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eNaito K, Zhang F, Tsukiyama T, Saito H, Hancock CN, Richardson AO, Okumoto Y, Tanisaka T, Wessler SR\u003c/strong\u003e. 2009. Unexpected consequences of a sudden and massive transposon amplification on rice gene expression. Nature \u003cstrong\u003e461\u003c/strong\u003e, 1130\u0026ndash;1134.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003ePortis AR, Salvucci ME, Ogren WL\u003c/strong\u003e. 1986. Activation of Ribulose bisphosphate Carboxylase/Oxygenase at physiological CO2 and Ribulosebisphosphate concentrations by Rubisco activase. Plant physiology \u003cstrong\u003e82\u003c/strong\u003e, 967\u0026ndash;971.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eRistic Z, Momcilovic I, Bukovnik U, Prasad PVV, Fu J, Deridder BP, Elthon TE, Mladenov N\u003c/strong\u003e. 2009. Rubisco activase and wheat productivity under heat-stress conditions. Journal of experimental botany \u003cstrong\u003e60\u003c/strong\u003e, 4003\u0026ndash;14.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eRundle S, Zielinski R\u003c/strong\u003e. 1991. Organization and expression of two tandemly oriented genes encoding ribulosebisphosphate carboxylase/oxygenase activase in barley. The Journal of biological chemistry \u003cstrong\u003e266\u003c/strong\u003e, 4677\u0026ndash;4685.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eSalse J, Bolot S, Throude M, Jouffe V, Piegu B, Quraishi UM, Calcagno T, Cooke R, Delseny M, Feuillet C\u003c/strong\u003e. 2008. Identification and Characterization of Shared Duplications between Rice and Wheat Provide New Insight into Grass Genome Evolution. The plant cell \u003cstrong\u003e20\u003c/strong\u003e, 11\u0026ndash;24.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eSalvucci ME, van de Loo FJ, Stecher D\u003c/strong\u003e. 2003. Two isoforms of Rubisco activase in cotton, the products of separate genes not alternative splicing. Planta \u003cstrong\u003e216\u003c/strong\u003e, 736\u0026ndash;44.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eSalvucci ME, Portis AR, Ogren WL\u003c/strong\u003e. 1985. A soluble chloroplast protein catalyzes ribulosebisphosphate carboxylase/oxygenase activation in vivo. Photosynthesis research \u003cstrong\u003e7\u003c/strong\u003e, 193\u0026ndash;201.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eScafaro AP, Bautsoens N, den Boer B, Van Rie J, Gall\u0026eacute; A\u003c/strong\u003e. 2019. A Conserved Sequence from Heat-Adapted Species Improves Rubisco Activase Thermostability in Wheat. Plant physiology \u003cstrong\u003e181\u003c/strong\u003e, 43\u0026ndash;54.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eScafaro AP, Gall\u0026eacute; A, Van Rie J, Carmo-Silva E, Salvucci ME, Atwell BJ\u003c/strong\u003e. 2016. Heat tolerance in a wild Oryza species is attributed to maintenance of Rubisco activation by a thermally stable Rubisco activase ortholog. New Phytologist \u003cstrong\u003e211\u003c/strong\u003e, 899\u0026ndash;911.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eScafaro AP, Haynes PA, Atwell BJ\u003c/strong\u003e. 2010. Physiological and molecular changes in Oryza meridionalis Ng., a heat-tolerant species of wild rice. Journal of Experimental Botany \u003cstrong\u003e61\u003c/strong\u003e, 191\u0026ndash;202.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eDe Souza FSJ, Franchini LF, Rubinstein M\u003c/strong\u003e. 2013. Exaptation of transposable elements into novel Cis-regulatory elements: Is the evidence always strong? Molecular Biology and Evolution \u003cstrong\u003e30\u003c/strong\u003e, 1239\u0026ndash;1251.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eSwigoňov\u0026aacute; Z, Lai J, Ma J, Ramakrishna W, Llaca V, Bennetzen JL, Messing J\u003c/strong\u003e. 2004. Close split of sorghum and maize genome progenitors. Genome Research \u003cstrong\u003e14\u003c/strong\u003e, 1916\u0026ndash;1923.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eTo KY, Suen DF, Chen SC\u003c/strong\u003e. 1999. Molecular characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase activase in rice leaves. Planta \u003cstrong\u003e209\u003c/strong\u003e, 66\u0026ndash;76.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eVaughan DA, Morishima H, Kadowaki K\u003c/strong\u003e. 2003. Diversity in the Oryza genus. Current Opinion in Plant Biology \u003cstrong\u003e6\u003c/strong\u003e, 139\u0026ndash;146.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eVogel JP, Garvin DF, Mockler TC, \u003cem\u003eet al.\u003c/em\u003e\u003c/strong\u003e 2010. Genome sequencing and analysis of the model grass Brachypodium distachyon. Nature \u003cstrong\u003e463\u003c/strong\u003e, 763\u0026ndash;768.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eWang D, Li X-F, Zhou Z-J, Feng X-P, Yang W-J, Jiang D-A\u003c/strong\u003e. 2010. Two Rubisco activase isoforms may play different roles in photosynthetic heat acclimation in the rice plant. Physiologia plantarum \u003cstrong\u003e139\u003c/strong\u003e, 55\u0026ndash;67.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eWerneke JM, Chatfield JM, Ogren WL\u003c/strong\u003e. 1989. Alternative mRNA splicing generates the two ribulosebisphosphate carboxylase/oxygenase activase polypeptides in spinach and Arabidopsis. The Plant cell \u003cstrong\u003e1\u003c/strong\u003e, 815\u0026ndash;825.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eYamori W, von Caemmerer S\u003c/strong\u003e. 2011. Quantification of Rubisco activase content in leaf extracts. Methods in molecular biology (Clifton, N.J.) \u003cstrong\u003e684\u003c/strong\u003e, 383\u0026ndash;391.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eYamori W, Suzuki K, Noguchi K, Nakai M, Terashima I\u003c/strong\u003e. 2006. Effects of Rubisco kinetics and Rubisco activation state on the temperature dependence of the photosynthetic rate in spinach leaves from contrasting growth temperatures. Plant, Cell and Environment \u003cstrong\u003e29\u003c/strong\u003e, 1659\u0026ndash;1670.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eYin Z, Meng F, Song H, Wang X, Xu X, Yu D\u003c/strong\u003e. 2010. Expression quantitative trait loci analysis of two genes encoding rubisco activase in soybean. Plant physiology \u003cstrong\u003e152\u003c/strong\u003e, 1625\u0026ndash;37.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eYin Z, Zhang Z, Deng D, Chao M, Gao Q, Wang Y, Yang Z, Bian Y, Hao D, Xu C\u003c/strong\u003e. 2014. Characterization of Rubisco Activase Genes in Maize: An \u0026alpha;-Isoform Gene Functions alongside a \u0026beta;-Isoform Gene. Plant physiology \u003cstrong\u003e164\u003c/strong\u003e, 2096\u0026ndash;106.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eZhang N, Kallis RP, Ewy RG, Portis AR\u003c/strong\u003e. 2002. Light modulation of Rubisco in Arabidopsis requires a capacity for redox regulation of the larger Rubisco activase isoform. Proceedings of the National Academy of Sciences of the United States of America \u003cstrong\u003e99\u003c/strong\u003e, 3330\u0026ndash;3334.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eZhang Z, Komatsu S\u003c/strong\u003e. 2000. Molecular cloning and characterization of cDNAs encoding two isoforms of ribulose-1,5-bisphosphate carboxylase/oxygenase activase in rice (Oryza sativa L.). Journal of Biochemistry \u003cstrong\u003e128\u003c/strong\u003e, 383\u0026ndash;389.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eZhang N, Portis AR\u003c/strong\u003e. 1999. Mechanism of light regulation of Rubisco: a specific role for the larger Rubisco activase isoform involving reductive activation by thioredoxin-f. Proceedings of the National Academy of Sciences of the United States of America \u003cstrong\u003e96\u003c/strong\u003e, 9438\u0026ndash;43.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eZhang N, Sch\u0026uuml;rmann P, Portis AR\u003c/strong\u003e. 2001. Characterization of the regulatory function of the 46-kDa isoform of Rubisco activase from Arabidopsis. Photosynthesis Research \u003cstrong\u003e68\u003c/strong\u003e, 29\u0026ndash;37.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eZhao C, Liu B, Piao S, \u003cem\u003eet al.\u003c/em\u003e\u003c/strong\u003e 2017. Temperature increase reduces global yields of major crops in four independent estimates. Proceedings of the National Academy of Sciences \u003cstrong\u003e114\u003c/strong\u003e, 9326\u0026ndash;9331.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAkYAAAE9CAYAAADu0K3zAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAP+lSURBVHhe7J0FoJTFFscP3Vy6u5FupEEBMRDFxkJ9dheiAoIKCAqCgtIlioSAEtLdEtLd3XHp+N78zt3B9XpRNi4XdP6+fdz9dvebb2bOnPM/Z2bOxPMMxMHBwcHBwcHBQeL7/nVwcHBwcHBw+M/DESMHBwcHBwcHBx8cMXJwcHBwcHBw8MERIwcHBwcHBwcHH2KVGEVGnpALl/66tvvUyUg5d+Gi793VgTXiV7tK/NzpU3LSvMClf/Ha8jOnT8rZc+d9764/XLxwRk6ciJRLvvexgXNnTsve3Xsk8tRZ3xUvoD6PPHHcyMrpqDeXosukuY8XO09//uwZU+4Z/fvMyRNy6ox9/n8PTp04Knv27pezZy/4rvwZF86fk5OnTuu4Pnf2pP4dm6C8yDCXcfbcH/0YOjyjMyPl4qXYHDE3Hi5eOK+25GIMtsTBITaQ4CMD399/i9+mj5ERY6fIlo2rZPLkqbJ6zWZZvniezFm5VcoULyIJ4sfzfdPg4jmZP3WUvN+yjZSq00gyJE/o+8CTdb9Nk3ebvSuZilWRPJnS+K5fGUd3b5eRwwfJ8s0HpWCRQpLIr5i/wpOl0yfK9yN+luWrVsnCWdPk4MXkkjdXVknwt7+78bB2yVxp1aKNxEtbSIrky+y7en1h84Kf5PVWX0ut2++WFFYEwogzp47L0CHfy7iff5b+P46SYhXrSMZkl2Thglky5MefZM26DbJ+9VKZNX+ppIzIIhnTpfL9UmTXtjUyZPCPsmTpclm/YZ0sX73GvLZIKSPLFsf2bpOJk8ZLmpxFJGXiBL6roWP/rg3S8eOPZOneS1KtbFH5pt2rMnXNcalZqZTvGzc+Dm5dLT8MHyajhgyWCXPXyc1VK0kyvzY8c3yPfNmutUxftU+q3VxWpn7fUXqMXCh1bqkmsSAqcvr4Iend5WMZMXuV1K1RVfzVVbA4tGuTdPzkfVm6+4xUK1fCdzVIXLog8yb+LO81/0Iq1L1F0qZI6vvAYcvKufL+Rx9LsXLVJX1ECt9VB4fYw1VHjKaMmyQJ0mSWDIlOSqfPO8rvuy9KnpyZZMKwKXL8ZHSPKZ4k8U7JkiWL5dR5fy88niSNf1HWrvxd9h876bv29zgbeVTmGpL1w5gZ8k8xpj0blkr7dh0kfd6Scvst1WX90nkyYdp8SWiU4OY1v8uCBbN93wwe21bMkmkrtqmXG5dIkiy5rDUEYN/+SN+V6w/pc5eR++5tKMnDxyn+hCWTh8r3vyyTl957V4pmi5DjRI0SJpb0aVJJvy/ayuINOyRX9qwypn8H+d9r78uWg1FyunvLCvnfU8/Kuj2npHb9BlK9SkU5snGp9Bk1VT+3WLdyjjz63PuyYtVG35XwIEHCJLJpy2rZtG23vq9U+16pUamk/v1vQa8uHWXnhazy0nNNJUOqBHLy9J8jYkkSJ5QjR/bI72s2aUSxcNlbpf4tVeVKojJhxizTXrt8764e08cOk91HDkuipCnk8O5tsnrtaglXEDlh4mSya8NK2bh1q+/KX3Hy4E6ZOnu2+ETvyoiXUBKeP2aczcUSefr6jQJfxsUTMnvmFNmx97DvQuwhXZa8cvfdjSV96uS+Kw4OsYurihgxjZUhex5pcEsNyZ0hmfT5YZQ0euY9ubd+VSlTsoBkzZxejh46IDu275AzF+NJqtSpJEvqRDJo6Gip0aCRnN6/Uy4YPzBVimSSxnjt40aPluI17pLiuTNLZORx2b5tqxw4fFxSp46QBNFCOynTZ5ZTR3fI4i0n5cF76kti87F34Yz5zQ5JYJRdkkR/qNItaxfLNz0HSuMnX5RK5UpKmTyZ5UzSNJIze0bp8MH7stp46FWrl5fIw4ck8mSkHDiwXw4djZRL58+JFy+BJIh3SQ4fOmQImCF2iRPJxQsXZO/OHbJzz35JnCyFJDh/UFq88aLsiZ9DyhTKJadOHpcLl+JJQvMIRw4flnOm7gnMrw8cOCSnT52UPXt2iZcwmXnmi7Jzx3bZtWefJE2Zyjzzn33is6dOyLat2+TwsROG8KSUeJfOyf59B+TcOfPvnp1ywJDIiDQR6uUyHUB7eYmTyMIpQyVf+UZSrngO352icOTIQdm2bZucOHlWUkekllPHj8jBI8ckXvwE4l06LwcPHjF/x5czp0/J4SMnTflHZfPWHZIkZYQkiueZvtwvxyLPyIG9O+X0eU9SpUwme3Zukx079xhrkFRSJEsiF86dkZ2mbQ6Y+17w4plrSeXw/r2mjnvk9JnzkihhIjHWSPLmyikpze/p1ZPHDpvf7JTDJ04ZGUlNjFzl5tipc3Ls4F7ZufegXk+Y4M/m8aKp815z3927dku8REkkedIkcv7MSRna7VvZeipCnnjiHrml1i2SM2NKY2DiS5o0GWTaqB5S6Z5npfEdDaRwhkvStscQqV23vhTKmVk+feMJWXs+u3zV5XPJmzWdpEufSSpXqyJ7ft8iVWpXMu0cT7yL52VQ17by25p1ciFpbrnj1opX8CIuyZ5d22W3kRFJlFifjcjl0YMHTF13y7GT541cp9S+8y5dlL27dxjD58mKFYvES51H7qxVXlKlzSBZMmWWBN5FI0cH5MSpC3Jo307Zd+i4pIkw/W5+fPH8Wdll2u7AwUPaj+fOXZDEiRJJPPOsFmeMPO4w/X70xGlJaLyB8+fNdxInlgOmX7Zt2y5nTV+mTJlcDptnizTyecLIxdbtuyR5ytTmXgnNODd1Mc+3fccuY6eTRPXz2TOmXNPPh47KRfo5+Z8jGedORcqOHebzw0clafIUktgMhnOHt8hX334r2cvUlnq33iL1bqkmqZInURmgbQ7t321k/Yx51rWy5XB8adTgVklq5D5P9mySLInpW2TL1OPIMSOzCZLL7lXTpVnLtlKoRAUpkDu7GVumnpR55Lh59lRyyXwfmT9p2m3f7u1y6MQZSZcmtezcuESef76FFKhQTYrkzSkHd66XtXsjpdFtdY3cHZIzZ89JoiSmPhfPmTY5JJcMOUlsCBs4a8bGkaNH5NixozpVmyB5aolvxs6OHVuNjB+QlKkiJFVEGuN8zZTjiTPJnbWryfmzp8zY3Gba6ogkMvVJkiiejOzTXvqNmiuVa9wiGVMlkmOmf7ds2SqnznmSxsgFOH86Urbu2G3uf1jGT1gkDZs0kSxpokjApUtGFg4ckFOnz8jx40dlq9F7yVOllJPHD8mGjVskcfKUps0S63ePHz6o4+vYqTMSYcb9hXNndXwhT4dNm+85cFSvJzBjH3nat9tc27dPvARWbpnSPGX0OHU4KsmMXCRKEN8830mVvf1Gx6EH0V/Tx/4obTr9IBUqVpEcZgxdhpGhg/v26BQq+id50oRyyLQt04O7tm2Wg8dOSepUPEM8I8NnjV7cJrt275fkKZBBxr1n2nunucZzJdLnumTumT1XPkmd0uhgMwjRT5uNvoQ/RqQiguTJvl1mvOw/YGT8kiRNmuzPMxgODgHiqogRyjdDpoz69/lju+XbfsOkUt0HpXyhjOZ6Blk7b7Q89VpzOetdkBZvfiAFKjeS/GlOSfc+38m6LTtkULcO8kW3wVKh9l2GWCWQwYMGSZl690vupCel09ddZIPxnL/9rLks2Blf6hpDEd1rXPHbDFm2/Yw84CNGRzcslcceflBSl6wrxXJl8H1LJKkxAgunjZYW7TvLpVQZpUylqlKtQinZvXKmdPvqG9llCEayRBfl+287Suv2PWTSpMny3dhfZeKgTjJ3V3ypVTqHPPfwbTJq8X5peGcd+anHFzJo1BxZMm2IfGqeP3fObNKn7yDZe+yCUaanpEvL12XE9ANy163F5bXH75eeI1ZLmdzJ5cWmT8r3o8fLqFE/yrGLCWTXqjkyZcFKGTesn/QbPkXq1KsvqZIa4gDOH5EXHrtHFu4+K+P7dZW5689J3uQn5fFHHpZf5y6T2ZPGyMftu0jGQhWkdMEc8m2792To+GWyYcU0GTJ2idRv2ETKFssedS+DHaasjoYc7tm+ST5u+b5EpsovJ9ZPlbq31pcNR5LLkWU/Satuv0jhovlkSLdP5dmWXWT35qXSollz+XXZNqle4Sb54Nkn5fMeQ2XCqP6y5ZAxoEcOyITx42T+9PHStksfqX7LrTJ9ZDf5Zdoy+X3uOJn0+24pmu6ctGrfT47t3ya9vx0gWfOXkp97fiBvtO0uDzz1lJzduULeeqeF7DREqm+XjrJk03EpVSSLvPHEQ/LNd+Nl/ZJZ0uLjthKZJLvUrlTcVxsDY4wGfdlOBv88Q7atmi+ffN5bSlatI0c3LZTu/QbKtv1EA5JI0SKFTf9HGTXPOytDeneVjEWrSpFsEdKpw2dyLqKQPPfM05I2/iF56cl35JYmL0rjW8ro9xXxE0vFqqUlkSEbYPfG+TJy7j558YG68mX3vnL/449JWmPc/wxPhvfoKoPGTJclM0bJ+5/1kLvve1C2LJosH7fragzaXvniozZyPFU2qVyioEwZ3ls6dB0sxw9tlgG9v5eiVRtI2ayePNTwDll9MpGUzppcHrv3bhlmjOPyOePl3ZbtJJOpQ5nCWeWbT1vKsOlLZavp3/dafSo7DopUr1JGiQg4eXiHvPXqq7Jp3ykZ8k0n6TVwmKTLmV8uHlorXQaNlD3rF8v75ncpMmSTH7q1kVad+8u69auly8cfyaq956Rh/RoydcS3MmDib7LBjOf3P+sjNQyRHDWgl0yc/bssmzlORk7cIo3uqqrlgbPH90hr43D8tmm3TBo+0Mj8fKl1S3WZP/pH6T9ktBw/e0ESJEwjpYvl85Eikd+mjZHWn35liMwB6d/lC8lSqpbUKZVdXm96n/w0f4Pce9dt0rvDh/Lrov2yeEx3mbkumUTuXCg/jJ1qmjuepM2QRTp+9Ios3bRHfun1lczfHk/ypz1l2rChTJq3RuZP+UmaffSFlKxeT/Yu+VUGGCfugjHkOfMXlePbV8jK3SekboUi8vQ9t8uH34yQ+x55TM5tmSsPNn5aUuYuKyUKRzkZq0wfNn34Yflh3Ez56bu+cjpjYVk780eZvWq7jOnXXvpPWid3311PFk/+SY4kymKIUWVp+cojMnrJTlk8/gcZPmm9keNC8k23bjLr981GvNJIjhSR0rVPX+MErZfm730giTIVl7IFUslH774p81bskZlTf5VxU9bIK++8bEhUFAE9c8T064vPSKf+P8l687sv27WSX6cvkE1rlkvnjm2NjlgnDRvfJQfWzZN3mn8iB44fk67t2sr244klX+bE8uyD98h3P8+WVQsmSfNWbSW5qWOl4rllUPcORm5nyeaV86V9x15SrGI1yRZxST5p/qEs3bxDenZsLhNWHJZGZoy0b9lcZq3aIrPGDpW+P0yRqrUqy6j+PWTKtLnG4b0kZatUuzzVvHbxdHmrVQ9JfHG3vPNef6lSNrU8++gTMn7iAhkz4gdp80lbuZgqj1QtU1AG9PhMZizfJpO+7yTfjF4idzW8TaZ931N6Df9VVi6YIM0+6SJ1Gtwho7p/JK+2/lLqmr9P714pHXv0le1b1kqz5i0kbd6KcnbbLGn/zXg5smWh9Bs8VcpXrytpU7p9RQ7BIyzSc+LkObnljofl0YcbS+qLe+X35UsM5UpgmL7IPU+8ItNnT5MKuUQ69xxsrlNkPEmcwJPJY4bK9HmbjFErLVVKlpDfF8yVHXv+eWooVY7C0q5LT6lT0tzUDxFZ8kj7bwfLa03ukHZvPCt173xYxs1cKQXK1ZTbq1eQOvUfkCcffVLuqF9dzqbIJJ2++tZ4dN2kTrWKxiM7KxEZcsld9eoYr+Oi7N+0Qr4dOFZea/2xfN62tTQw3yldtZbxkEpKw4efkpeeeVJqlTckznhTqdJkkvq1a0nSMyclf+myUrloFslRvKb8MPQnqZgjpQw098met4jUrl5VNq5ZLavXb/Y9MfzKeL1ZSkmzN1+S4vkzyypDEHLdVEIqGQWdOU8p+W7oUHmoQl5Z+dsq2bpsgvQdsUBafNZOPvmso1TOl854lP4LNT3p36uHbN93SUrcVEZK5c0m0ybPllsfeFm+av++LJs6TOZuuCS9enWRmpXKS4VCBY1hjS8vffC5jB/aQzbPHiNLtsUz7VFaUqaMkAEjJ8qT9SpIl3ZfS8YCZaR8+Zsl4bGtMnXhapk+abYcOiXy1oft5OEGtWTjKkOSNq2SkjUbSNs270nOXLmNsa0tCY13msR0ee8On8iFdGXlg7ffki5t3pCR/TvLyv1JpE7VMsbjTW6MtyGfrz1oSOgcMY7mZexat0C6D50sT7/bSlp17CqlUx+Wdh2/lOJVbpOH6tSRUjXukleaPiIRfmsy4pn/LhnhW7t0gTz35CMybUdq6d/9CymYOamcPRspx84lkGQ+b90fSZL+cY+RfXpLnUaPSf26tSVXiqMycMQs3yd/4Oj6RfJl31/lzfc/kg9bfSwPGnIReWiPdP26uxS/tZG83fx9ee/VB6Rbu/by25IFhlT2kgdeaSlvNmst99SvqguwsxQoJ5VKFTD877xkK1RayhXOKekz55I+Q0bJK40qyrxpv8mBLatl0C+zpEmTJ+WNV14SuXBBSlYsIyksuTZYN3uULNp8QF5//w15+5l6cuDwXqllDFjXzzrIhfgZpEyJ8pIvbWJZYAxwvbrV5Vy81NLKEJSeHd+WFatXyF7jhbdt/7VkzlZUKpapLCnOH5JJE2fKprULZc/pS/Lsux/L/+6r5istCuO+7y2Lt4p89MF78mX71rJv5XjpPWKe1H+yqVQuX0Sa/O9FefL+W3zfhkgZoti1m9xs2vWNt9+Vl198Sk6fPi0ZchaQu26pJBcvRk2Wz5ryi5xMkdIQw0+l8R03S507G0qJsmXl+Zdfl9K5U0vugmXlGVNGiTxpZPWSaVLYjO+iWc31ImVl0Ihxcn/5zDJrxlq5487bpFj+4vKC+V3ZQjlNs10wr/OSKWdhaflRS0mb6LScuXBRjh49K3c0eUrurF9JywfFK9aRXHkyGaNfT4aP+kXKpNgnvQb9KgULlpQaFW+WnWsWy4p1hyQRxJSo3SWjt1LmlXfeeEXKl8hvCOxcuZQsnTzcpJHcVK6avP/WY/Jj726GuMaXqlVqSuG0yWX2zFkycnBPWbglkXQwDk/rt1+UEvkyy7nzfyxWT5Yut+mvKhJ5IZW0+ehzGdi1haxdskMaP9tCfhnSTQ6sXidbDBn+tu3Hkr5IXWn21tvyRfOnpeeXn8o+ySbVyhSS1GkzS6/vR8g7TWrLoukLZf3S2TJ46HT53wtvSpv2n0mlnBek/Zd9ZPyg7+S3nfGk9fvvSZuPP5Vy5lkmDB8kM5cfkw8Nkfvys0/k/M650mfYArn3gfulnNHZrxlHOHPKqIgV2LtuvqzavkPKN3hMPmv5kpQsUUmKGQfIy1BAhk+cLO8+c6f07dtfFswcLz37j5H8BWjPqnJo62qZNvIX+arvGHnx7Q/kg5Zt5OHb60jixMmk9m13S5bU8eXC+bPS0zjSu44kkmpGFxdIkUhmzJgly5YulA0Hd0v9B56U9978nyQ3zq+DQygICzEqVKaKnN+1TH788VeJFz+enL14zFw15CdxIilTvJAxQtml7m21ZO+BPXod79G7cFb27TpgCMNNUqJ4YXnktVYy+Nt2kj1tdK/8r0iYIrWUr1JFMkSbcz4RGSkp0mWRL77pJzOnjJSMlzbIU2+9LwcPHxPPu2gUbxSJ8Iyxzp03txQoll9y5cou58+dvzwlgXKOHy++IUa/SaTxhzKkiy+Js5aQjz/5SLKljprCuOhTXN4l/o36nWc8J8/8ndAQv/gJk0hBc/8cWbMJQYb4idNJqdKlpHr9xjLml6FS8aa8+huQICKX1ChfQL7p8JkcPHZJLnrHJL4hjYmMQihYIJ/5O4lkypRai5k/a46pfBbJGcEvk0mWtNG679Jx2bv/jBQrUUIKFy8u7xhC83XrFyVpovjS6L6HJKcx8PN3x5N82dLq15MnSizpDSkrnjOTlKx9l5TOHiG7t2+TJMlMnxYuLbmzZZU0huDGS5FKShQtKBVr1ZXBI8fJQ/WryCOPPSbzR/eRCpXvkB2HT0nVeo2kVJqL8tDdt0v7/qMltVGWiVOk0Gmf+HJOpvy+xbRllDeep1INyZUkkaxZtV6SJY8v+fKXEHo9IlUySZnowp/WgOxYMVVOxk8kGdObNjCN0KBhHdm6Y4eYauo0w8UYFoywfzG+6YcSFavLnVUryPlDW+SsZyplED9BcmMQT+l06l+g/SlyatdymbJyh6xZOMYQkukSkS69DOnXXaL2Of6B9Rs2yF5JKIhshmwFpPUnH0uG5Bdl5ZrfJWPGqAXxBUtXkiSJImXJ9LGGSHqSN39OvZ4qe0ZTnSjZSZLCyHHUn6Y9xBiLqEXY6dKYNjRtlyp1esma+oLMWrFGDh05KhGp00nOTJn0OxbpsuWV0wcPyBLz3IeOnZL0OUtJGu+IITzxpVy5ElKwZDnp0GOQtH7lPp1GzZMvt2ROn0SSZ8po2lHk5IEdcvR8BqlS8SYpWvkWYziHy+P315P7HmgsS37qYYzzbbLu2DlfaVFYYPovXuKMGuFNkeMmKZk3j6xZvIyOMQTkopw3ZM8fB/fvkw1btkqO7FFtkyxDetNZpq/M30zR0HP8/cwLr8iUr9+X2g++IAeOHYmaSjSk5tLFC5I6c04pmC2l9Ok5QI6dNkTn0mmhlFSpE0r+fFELoDOkTWR6JWq8X7x0Xs5f/LOMXDBvKzS4WyrlSiFdOnWTWatWSeZCBSS1n+qhOxJHpDP6Ka/kyJ1Lkp89JknTFZTyZQpLrfv+J6OH9peyBdLL2Qum8WjA+BFSp3pZGdilg2zdc9pogxOm/HNyweiWi+a5aYode/ZKmZKVJF+e/NK+x3fSpeWTMnfe75IqfR6td0Sa5JL8L1FJ0ypGTvIXzCtpUyaQREnTScGc2SV9pvQSL2FySRE/ocQ7c1Amr9ghWTJGyVbR2rdKxnNnZOOGrZIiZfzL8pQ2IqmkTHBetmxbL1uPGrk15NNIn1SoUk32rlgpU+bNl4QZo+SqUu275d3Xn5Plq1bK2fjpjCMbTxJlyCelCxeRtYuXaLteNH3s/ckxEylT934pleqw3FqhvAyb/ZtpxOSmLRNJ9bosvk8stereIQkSnpIdm9dLooj8UrFcYanS8HEZO+pHyZj0hGw6dVHSJY0vaTLmko8++UQKZo0w9UwoCZMkVnnavfeAlCll2jBvAfmi3/fy0euPSoN7H5XUu5dIvQYNZdjkeZIq+R9EzcEhGARMjBIZsoPRSWAGZBQ8+a5jG1kTmVbeeuN5SZ8qkfF4zsu50yyujmc+jcL+PfukaKE8psR4+vskSZNLKjNQly5ZJOfjpZD8ubPIit+Xyo79R32/+AMJTZkJzODw2RFDqs6pkj3r51mBrRvWyc8/j5WLCRJLpVoNpW2bduId3iPHI08riYmfMOpp4pt7oSt973QqxuesypGjhwxximcMTT7ZsW2XjB5jBrfBnCk/y7w5GyWxIRlWFyRKlkS8BFE/PBF5TC6ZekGwEiU0/+qSUkM+kiSXTcYj33fwhDGK+WXbprWybuMW/QxsnT9e2nQaLM83bykVS+bSFjth2o458ni+xoOoJTSKKWPaVHL02EZZszOqbU+dZWu6n3cUP6kkT3FRZs1eIBEZc0j29EllzoLF5tlOyYw5i6RA1Tsk+YFp0mXAWP26pkC4YCN0Z0yfJZDs2Q1p00c3Ss/8f/KICDm1c60s23pI8uXLJ+f2rpNpS1aLpM4rM42CfLhWXvm4+duyeNsJaflFDxnZq4NMHD5Qhk2aJamNwceoJTTmPX+KZLJ0xQxubAo+J5eMksyVr5BEPb6voqYvEiRk3YzvrUH6nMVl347dsnrVBn1/5swFyZY+jZiqabvEtx3nDy+h6W8jY8lTyVMtP5eKhVLIcy+8IPuPn5UkqbPIgw/fLmN//klW7fxD1k6fOCyDvx9kqLDI0NGj5bamb8v7774jz730hvT5sq2c2bVCvvtlcdSXfcicMaMk3LNIfpppyIDB7FHfydKtJyR1omSyatU6vXbWjIUMaQ2RyZNbjh09Jmt/X6XXSSlxEQttwJqmBL51VT63wfc38hRfkqbPJvVuqSCrJo6QESNnSvM2HaROhQL6HYuMBSoYByOPjBrSV34/kkr69e1uxk0SSWhI2cxZiyVbnnySPtl5mTbHPKshRraR40NcTdnJI9KbZ9opCxZvNEa4oCQ8e0DGj58uKY2nP/m3pdKkdkGdulmz7w9ylDdNetm4Zrac0A0Wl+TspYuSp1ARc9MEkjChIdSXdUQUkrBe5OQx2bBhe9SF82eNgY16DiIvCc24RPQSGNlavHyp1CuYWl5v2daQD0Oyzl3UNlo2a7x83WO0NH3xFalwU1Zdb0iAEafkj3Yz7ZkoUZTRRsDiGVk2H7F+MWEC1lLxraTy4uvPytCuH8vs33ZL7Zq19LcW3A0nz+4QT5I6pWzd9LvsPnBG8hfKLzs3rZKVG3dL0sQJdCPECaajW3eVe194W26tVkwD48dPnjBl0TYJ5Sxh0EtnNHKYIXtuQ3qTyZxZ8ySZIYSb1i6Sk+Zr50wdLxjd5tmB70PCREZfGZIXdTWB1lVbLV5CIzueJEiZVvImji9LVkVFNT3TrvGMU5I9V35Tfb7tu59nJMrIRKb0WeXsnjWy9WDUYnZSDmQxRKN4nmyydu4vsmr3cbl46qj8asaxxE8vuzbMkyNnotIcnL14PqqPzS3hhN65U3IuStUpjuzaLR/1Hys/fPOR/DqwgwyZvlbXu506EzVOTxu5z5IxgyF2mWTH1hWydVekac+CsmfrKtl18KxEHF0hI6ZH6dy5Pw+WOWt2+/SfkQ/W1F08JYuX/SaZc+Y2zlciWTB7lmzZd1a++3mCdGnWVHp07yrzVq6Ts6ci5ejxq9vg4+AQHVe9XR+cOXNKpv4yTL4bPs54U1mkasXiukBz5axf5cfxk+XUhdMyfewU2XMmqVSvVEJmT5sim42wR+5aJfMWb5a333lPTm1eLN179pek2W6S22pXNYOns/T+cbTMmTlNJGUOqV+v6p+25J86uFW+HzRQFqzeJeXLV5VcWdLI4XXL5bmnn5KIEjWlULY/Fv4d2btZvun2hWzaf0HOHN0ro38aLfnL1pJHGt0qKxdNksHjZhpSlFrWL5kqE+atlHJlq0uBnBll18bVMqjvENlzZL8smTdTthw6K3UaPigX1k6T7r0HyTJDAFZsOyL/e+4RmTl2pIweO1Oy5skj2VKflQHffy8HDJmav2CBrNtzXArljpBfRw+RDZGXpHbd2yVP9kzGix4vn3TsYUjgfNl58LTxbG6XNCmivJqT+3ZIvwH9ZfexE7Jm2e8yf9F6SZIikcyfOVGOSnIpnT+bDOj/jeyMTCBNHr9fJpv2Hz1ujpyN3CnDjTJIlCmn1K5SVheLo4DTJPOkxxdt5KeJ02X65KmS19TxyJopps1HyUddBkji9ZOk7dc9JHeZGpL86A75btgwSZ2zoCyZMlx2SnZ58X8N5Mce38qvyzZL9Tq1pUDRwnJg9zpp1fwDWbjkd5m1bJs0alBLendvJRuOJJMSOZPKnkvpJYsckj79B0v+kuXlxKGDUrFqZVk/d5KMmLJAKtdtJNXL5JIf+vaVfQkidMrucIpC8vyjNWXAN91l4YbDUqNaaRlvnmXK0s1yS716kiVD1Nb6iEzZZeuiCTJx7lJJldiTkePnSZNnXpEcqc/Lt12/kDXbd0npyjUlqyFL2PtLZ4/LnBljpEPnIXIhcWq5pU5NuevWyurJj5u7XPIWvkluv6uBLJo+Rn78eaJGIrZuWCtTp82QVPnKS6I9S6TZp72lYPHKUrNiMY0AHj++X/p1/kaWrtkoFY0BzZZBQ3aSJksm2WhksWPbjrJmzSpZsvG4PP14Y8ma4oL06PedJEoRIbOmTpLcxarKE088Ietn/ywDfxwhx8+dl6njfpLdJxJL5VJ5ZUjvb2T7qWRSvVwe6fttP9lxKqHcXDqPfN9nkKw6cNZ434VlQJ/BsuvkBTF2TTZtWCnn4ieTQvlzRxlIgyPblsnHbbpIxuw5dCfn8hUrJVP+slIk/UVp27qlTJm7QKZOminlzPMvnTFSZizaKhUqV5FFU4bKuGm/S527HpYsF7fIRx+3lQVLfjNEeoU0vNfIwrftZdG2s1KsUDY5cOCUNH7wXjH2SJEtW3qZ+fNQWbTHkJMdv8mSHWeMc/SsHF05X3r1HyR7T6aUMiWL6C5BkDx5Sjm2a7V83XOgnDx1yhC2abJx+xEpXby4IdODZPGWg1Knfl354o2n5IQhR+kSXpJLhsjeXbuMjDD3W7dth0REpJGpI4bLjhOnZNu61bJo5VZJbkjLmJ/Gy4VUGaVknlTS65v+sv9iSqlX3cjU2B9k7toDki5VOvlt8nCZuHCj1L7lVsli5CV73sKy0BCtvBUbyr11SuszWuzYslq6fvmFHDJ9dGeDupI9Rw5ZOLa/fNlzkCyaP1N2HvakRo0yRq46yrrD8aR66SJm3A+TtTv3GXlaLzOmL5VMhUtKvnSJ5Ntu/eW0l0TurltOun/RXkaMmy5z5s2WHCVqS4MqeWVgr26ycMVu2bllpUyaOV2yFighZcsY2eNBzh83uqmXzFi8VW6uWl0WTB8pw6cslkqVqsnulTOkx9CxUrx0dalX2dyn3yA5nSKdLJ80QuLlriKP3lVcunf+WtYfuiTVKhaSYQMHy8JNh+XBBx+Uc7uXydDpyyUi0RmZNG2uNH7qWWnYoJIM6t1Dho/4RRYZfXYmeX558YkGMm/cCJm5+ajEO7Ba5q07IK+9/bKkvHRYBvbvKZtMn5esWkXS+lKyzP/5G/m071SpXr2cbDLOTMP7Gsv6RVNk5PTVki9bahk9aqSUr/2APHDvXbJy8mD5/Ju+8tuC2aZvj8kzrz4vh3etk3at28madWtk0brD5nv1Zem00dJvyBgpW/0WqWGcxw6ffiyjfp0pc+fPk6JVbpONk/rLTzOWSsECheTYsUip26Ce9Pj4LZm1/KTcWstvHaGDw1UioNP1z549ZYz3Utm+54AkM95i2TJlNa/Esf1bZfyE6ZIiawFJG/+U8X5SSo3q5WXbhuWyeOUGSZUqtRQuUVoK584uu7esM97gGkmdMZtUrlRedm9cLpOnzzUeWWa57fbbJVNE1JSHxckje2XRkuVy4qwnhYuWkEJ5s8kZQ2DGT5gipWo2kHxZ/8iFdOzoEVm9YrEcPH5OLhovyJgRqVm7pmQ0inn/zg0yZdYiKVGmkhzbt1UOHjsphYqUkaKFcsrJA/vk1wnj5ESC1FIkVxbZf/SElK9cVVJfOiHjJk2Qw5Hxpe4dd0j+7Blk6/qVMv+35VK+Vn3JmTq+jBs/To6fTyD5c2SXYyciJWfOzLJn5x5juJJI+YqVJWu6lHJg1w6ZPGWyHD0jhizdJkXyZvU9sYEhk7OnGwK5/7QaugO79krOvLll954dkihpSimcJ5esNkoiXsIUUsWQjSO71srEGQsldbrMkjjeWcmQo6AxLDdJSkNQfTeUZYvmypwFyyRr3mK6Rmb78tmy0yj46tUqyb51S2Tx+i2Sq3BpOT5/jDzfY5J0+fgN9RrLV64hGZN7RvHPN0btgpQpW97UK7OcOXFYpvw6QTYfOCE317hFyhbLJ4vmTpW9h09K4vgJpGCZmyXi0jGZM3+hJDZkIGXajFLJGIoVi3+TXYeOS9GSZaRQnhyybN50Wb1tv0SkSW3aprqkSXJe5puyjp+LL8UL55fdW7fIgVPnpFy5ypLTb6fL0cO7ZfLk2bqDKH3WPEbJl5TD+3fJ4iVLjLeaUPIWLC6F8+dUL907f1JWr1op6zbvVUNcpmJ5yZwuQjasWixLV2+VAsVKStmbCsqRg/tkhiHux854kjZ1aslsvOvyxohvN/27bN1myZK7sFQqU1RDqnu3rZdlv6+W05fiSdFSFUz/ZYt6MIPII3tkwrgJ5rk9qXXr7eazzEb2ThpCNFX2R16QVBHppXKVSpI2RRI5vn+HjB03UU7GTyWZ0iSWpKkzSb4cGcw42SAXk0WYvs4k69dtkEumrwvnyS6bN26ScwmTSe60iWTwjz9KpnylJCJZfCO/W2TaqiPSs3NbyZg2aq3U8qmjpG3fmXLPXeXllCEdyxf/LknSFJT2n7wqc6aNlyUrN0qhkpWkZpUSsnjOTDl8wpNCxYrK4d2bZP+h01K8TAXJnTmpTBg/XrbsOiTla9STysVyybL5c2XjvhO6nil3oWJyk28q0GLrmmUyZ/FqiUiXRgoVLWXGZ3bZsW6VLF+3UeIniZDipYpLzix/bJCINGN3ysQJcvBkPEmXOZ0x/vGl2E1FTRuskVMX40m5ihVl6+8LZO+Jc5IocVIpUbqM5MqcVhbPnyWb9hmjd0tNWbVgmpHFc1KiaH7ZvGWP5Mtlxtz+vZIsdTrJmymdrN1gyk6eWm6uUF62rlssq7ccMYa6imxfu0z2Hj8vpcqWlbzZM8ulC+elxfMvyIOtv5CS2aPIrsWBvdvlt98WG18jQireXNnoueTGQdgkEyZNl5NmvN/W8F7JkOSMzJ+zUM4kTCkVyxWT3YawLl23WwoWLSJHd+6Q/KVvllwZE8uUCRMkde5iUq1MEdOeM2T+srWSs4BxDG+rYzSUJ0sXzZEFi1dItixZohaKZ88jZUqV0DQjF85Gyty58+TYyYtyU4mSqsf2H46UQoWLy8Vje2TD7n2Sr0AxKV7EjMlZk2Tr/khDHlNJ5aq1JfGFwzJvwW9y3pDoovlzyVYjTycuilSuXN18dkQmTZkpCVOkkqzZchonsZSWt2XtUpk6fZ4kTZdD7rj9DkmTMoHs3LhKZsxfJqnTppH8yEDBPHLhzAmZbYjt0Yvp5E7jzCaMH0XR921eIbOWbJIURt5TpM8mNSoUkDeaPiQHUhWVJ+6uK5fiJTXjweiLZAnliNHD4ydNk2Nm7NS9o6EUMPr1lHFCJprxtOfYOalRp4EUK5hNVi+ZJ+u2HpCCJUrITWbs/TZ3uixcvsE4OSWkQd2ast3I4OKV6yVZygjJmC2HlCtZVGZPHCdeytxSs2qI+aUc/pMIiBg5/Lvw0+ctpMWPxuueP0ySwyocrktMGN5NPvzyF+nweQcpmDOdbFo+VyauOSzvPP+UpPGtp3jvsVqyNVUtafP2E8bYXpBZ48bLqWTZ5bmnG+vnDn/gzNED0r/fANm4a6ucS3WTdG31ou8Th7Ajcp889dj9kqXWM/Lpa49fjnA6OFzPCGgqzeHfgzORR2XJ6rWGGl+SnPkKSXbjvTtcn0ifNpNE7tokY8dPkoVLF8neM4nl1aef+NPmg0zZssvKBdNlxuy5MmfOHMl6U1V56vG7NeLl8GecOnlCpkwcI3tOJJAP339LUif7Y3efQ3ixe9tmWbd5p7E0KaVEqdKSIumf1505OFyPcBEjBwcHBwcHBwcfnEPp4ODg4ODg4OCDI0YODg4ODg4ODj44YuTg4ODg4ODg4EPIa4xee+01Wbx4sR5W6eDg4ODg4OAQlyBlycMPP6z8JBiETIyOHTumqdr9T/l2cHBwcHBwcIgLQGuSJUsmKVJw1FDgcLvSHBwcHBwcHBx8iHNidODAAc1+zOGtZcqUkdy5c8uuXbske/bsvm84/JeBeJ44cUJPRo8fn3PqLukLMH2bOjWHy8YeCMkeOnRIkiRJouXxPJRvhw3lI8P8mypV1NEX4UJkZKScO3dOEiVK9Jd7M0Z4VaxY0XclbrF//345e/aspEyZUqPHjGfOPqPtOGPvasDvFy1aJKVKlQp7Wx49elTb094XT5Ln+yfs3r1bkiZNKunSuTxfDv8MO2bRVYwBZCfYqMU/4cyZM7Js2TIpVKjQFeVz/fr10rt3bz2SqFixYr6rVwdmg9B1jGf+TZ48udbnaoHOnjx5so65ChUq3FDLbeJs8TWN9tlnn8nNN98s06ZNU+PXqlUrKVmypEycONH3LYf/OhiUDG7Od8qTJ4+8++678uOPP8pjjz0mNWrW9H0rdrB3714tD+Uz+ufRkjdvXrnlllukf//+0r17d5Xd77//Xo3nW2+9JRs3bvT9MjxYuXKlOguNGjWSI0eO+K5G4Y033pDatWvrs/0TUE6xia+//lo6duwomzdv1nn9HDlyyPvvv6+k6OOPP5YBAwb4vvn3mDp1qtx+++3y1Vdf+a6EBzzX//73P1mwYIH+W6lSJSVgVwMM3euvvy6//RZ1sCk4fPiwbNu2TU6edIeUOvwZjNnGjRurg9+mTRu59dZbpXz58ip7gQL5ImhwpdjFvHnzVAcNHTrUd+WvQGeOHj1ak77u2bNHydrVgmeuX7++3qNDhw5qo68WOCK0wzfffCNPNn1Sj84KN3bu3Km6N5A6XTVMo8cJPv30U3rba9mype+K5xmy5L344oveF1984bsSdc00su/dn3H69GnPNIrvXRRM53nG8/S9ixnnz5/3DNv2vYsCvzHGx/fur+C+Fv5/A56BZ/ynch2ChyECKi+DBg3yjMLwjGHy7rnnHt+nV4bxdDzj+fje/RXcKyYgI/fee6/34Ycf6vutW7d6xgv0atWq5e3fv1/vaQiS169fP/28a9euniFNVyzLGNgrfkZZyLI/eM+zGwXrlS5d2jMkw/dJFL777jvPGGzPGGnflSgg19zPYtKkSV7RokV97/4A9/b/Hu3AM8YExmD057MwxFDvv2/fPn1vCKL2U8+ePfW9MRSecXa8cePG6Xt/2L4xHra+X7t2rdZpxowZ+v5K4DlPnTrlexcF2of78fLH8ePHtc/atWun75955hkvY8aM3uzZs7Ve0b8PaEP/sTxy5EjPeLyeUcTemjVrPGPsPENWPWP4fN9wcIgCsvzUU0/pGFiyZIlnnBL9u06dOn+yOdgM//GGvPnrIt4zFt577z3flajxgp2x42X79u1a1rz58/S9Bd/zt1GFCxf27rzzTq9cuXKqD64WjM2GDRvq869atcp39a9jzdo/+1zAkDEvbdq03ieffOKNHTtWxw7gntFtNuBe6CN7Twv/NmHM29/OnDlTx2D16tW9iRMn6rVwIk6m0mB6ppOUCY8YMUJM5XyfiBgFKzt27LjMso0h1BBc8eLFNUpgSJOsW79O0qdLL6azNMLUvHlzSZMmjXqaW7Zs0ZA8q9EJMVqYhpQffvhBw3pGoJTRfvDBB1r2qFGjZPXq1Vpu0mRJpdm7zcQYAjGdKqZz5djxY7Jl8xapWrWqmI6TWbNmiRF0/T0Rjc8//1yM4dTplmbNmkmWLFl8pTqEC0RIvvzySxk2bJiGdCMiInS6lbbu06ePekT0d86cObV/X3jhBfX2O3bsIFmzZlNviXvQn507d9b+JRRNlMcMfnn11Ve1Ly2MMZSXX35ZvTH6nQhBgQIFVF569eqlMvPcc8+JMbySLVs2WbFihdx9993a/1y3oAwiTEQZiO5UrlxZyxo/frzWJXPmzHpv0Oy9ZlKubDn55ZdfVFbTpksnfU3dCIETRbXhcqatevTooXXgGQ2R0GgLY4DoGnKIp7Zhw3q57777daqPyE2mTJnUy8yZK6fEM//RZtSPsYFXyZQd017ck8+I9FAm5eEtvvnWm1Kzxh9ROq4RtWKsfvvtt3rt7bff1jFKnxilrdf+98wzsnHTJpkwYcLlcDoR427duqnHRz2IxBmlp/UgOsg015gxY7TuRLyYmiCiTH/xrExxGmKsHvkXnTrJSdPXPA/XavpFEonsMY4XLlyokSzkwpBZ7Sv0RMGCBbVtDFHS6B9TBfQZ/fX0M0/LXXfepWMeWSMaZoiy6pZdu3YaWcslffv2lQQJEvhKc/ivwxh3ef7551Uuli5dqjLGmK9WrZq0bt1axxQRyw0bNmhE1RB2+e23RUbWx+pYY9qNKDVR2BYtWqh8vvLKK/L444/rNWwUY4FryCxj9+mnn9bfErVFLzJesaMvvviiyjljqEuXLjrWkVV0w9UA3YbN/fnnn9WuMgawdYw9xtp9992n9WL8YE+ZQrz//vt1KpwoK3qvXr162ha0A8+HjmLsYzsNSVJdkSBhAilapKg+H+OLKBnLBNBj6FWeHx1rSJ1G7YlevfPOO5I+fXq54447JEOGDGLIn++pw4M4mUr7/fffdWoAI2W8N71Gw6GQMBRMH2DIDFvWcKLxSNXYQF5Q8v379VcFRoMYz1SWL1+uSrd9+/ZqxAxTV8HxB983Xqt2MuHB6dOni/G6VYAQWASV8OeXnb/U72FgMTgIcp3addTYdu3aVY0jz0qnQ4YwCChflDnhdoTEIfYwePBgadu2rZJqQrz0K+SY/sfg8W/ZsmWVAH/66aeqOBg8kOw2H7dRpUK4G2LBvDfTZRhxBr4/6Hfu7b8+ht+imBiUU4wBZz0NAxYgO5A05vP9gSxijLNmzarEi3IpC/mC9CN3KBdI0sifRspZo1yaNm2qcv/cs88qCYzuu0BwIOc4AihiFBLPy7PRFtzrp59+MuOorOTPn18VDAqLz5DtLz7/QpUPinmTISwoP8YSoW/qhoJC0XBPnh8yCEnp26ev7wmiQJoOxkWRIkV8V2JGzly5dLz5TwcePHhQlScEhOeCHFFnxhPPhqItXLiQKmf6FWWPIqVtqD9tDQljjHY1Sp+2LVyksGzfsd1XQhRQpoxXSBGgLTEOKGz0DGSbsYws4aTRrqzHwOl69513tVz6HceM8c39+IzyT506re3v4BAT0EUsF4EMoAMgFpB8iA/OEE4BtpAxxzjHwcBpG/HTCNVhkBzGBlPpyCiOAboMQoTcQtjRNyuMPkO/8B3GPtPROBgDBw7U7zA2GHuML/RUMMBpws5SLk4LTii6k/HKM+MgEfBAl5w+c1qqVKmidULHsv4Sssgz4YQyzrDtEBscHWwuziC2mGdlDEMCGWPoSe4PEWMMsowCvYPuxZbDF9Bd4UacrTGiAVBSVumjbOh4lBINgcJbu3atKlAUPoTk3PlzSqYAXmERQ5hQejDQKVOmaMfjldOgbNXzB0KGgaDDIE90CkoXA/rkk09qQ69bt06/y33ofBodxo43C2vlN8zplihRQlkz36OTuQ/Pz7NhbBxiD3gjDBAGPIAUYLgxshhpiDZeEkYUUlSwUGHJb2SHfpw7Z65GkRioEAaiDRAfZIj7+AOZgxghAxbIKp4ZxDqtUXYA2QPck/7nGfyBwX3WEByIPkSa7/PCuANksUGDBvo3YwI5RoZyGTKBUkQubV39gSwDFBblIu9169ZVhcpn1J9/+QwigKcFOaAMxhKKCLJIPSEnNxn5ZiE3ZUEAUGqgdOnSqpi5h62rBaSBMWAXNF8J1AFQlgXPhnJFsaPoGc+MUSJKkA3a5I477pROnTqp4mPNEh64jSgz/jAGPDt9hEOyZPESHasW9CvtQN9Y0IfUDRmCkFInFC33oG1QxKpbDNmjz3DGAPKDLkJOojzY7IYwxdM2cHCICTbiggw98sgjOk7QKUSMcDogScjP9h07pYQZ64wHZHPK5CnqcPFdZBydxpo4xhpOHboDWYVwKYxMM8bsmIKkYL+Q1fNGvm+77TYd6+gUbF0wQC8wPhmLnDsPoatXv54GFxizkDpsI4GLRQsX6ThCN/EvUWy+j91Ex6CLIHBEx+zYxEkkOsTYS54suT4rOgC9xP2ZYSptHJmTZtyzZpDIPLq1ZcuWOi7DjTghRjQg5IUKoXwACh2FCMulQ2kYvEgEhdA1BqNUyVKXDRgKzhoMhITv8h6lRSP/XbjQkjE8Qf4mhE/n2XAc16PD/oay+Jt/edlnKFa8mD47TNch9nDTTTdpiBiFgVLBqAKmUMCvv/6qAwfFQN/EN31kOswoiPPaz1yzfQn8+9IfCc2g5rv+EQHeo9zuvPPOywoGxYDsgZiiBwxqZBbPCI/RlsWzAO55mXCY6/Ye9nOL6M/nj+j1AVaGub/9rf3Mn8jYa3zDlsm/0Z/PvwwLFF9MhAn4P6/9nO9boGSJ6BG1wUgQjeVzyrG/JTJIRI0pOfQDhJGoMsqZha04JShSvE/GLiRr0MBB+luADPCKqV8ox9bJyoo/qJc/IGLcBwWNfmFKAe8VPeXgEBNw0NBL6CqAvCFnONh2LKgMmlcCM14vmDHH54w9f/kE1u7VqlVL7QyEILqMMm78f6fvzf2QVV5M22F3gwHPBFljqgvyxUzNd98NliSGvPEckDiIoP+z+78A5A1dyYtnu2SuU1+cOuuIAa4DWx8L/du8KANySBSb6e9wT6OBOCFGeN1EhwjvwRxhwsAqbCqNEYFtshYDtsy/GDyrwFBSCAvv+ReFRbiNtSFEmpgO8AfKmQa10QHe81s8fLxROpX7A/7le4Tz6USUIgrZlkXUwd6rRo0aWo/vBn2nUSOE1iH8sMaNuWcGAoORqRTINQMGgsJ7+o7pKgYqMkF4l37cvWu3ehxEUOhD7kf/8S+yEN24lyheXL+HBwT4Ln1O+fxNpGfosKG61ggSgvwgfxBzfyC7hLgZ/EQ3kB0ik/5kinLAOXPfihUrqNIk1L5i5Qr9Ht+xY8SCOiOPyKWVZf7mXlZOAcrl4oWL6o1BMvid/72IhFDenr17L08nQj6t4qbO9l4XzX39wTjF66NOFjwDoC0s7DU77QgIwxPRg1zg+VFfWw8UI58xhUZE780339TIER4yXjHThL8t+k36mXZduGih9gffJepDhMgCGeE3jF8LyrDtuWfPXiVoTKtRR/qRsYy8IEf8FiMA0BMYFWvkHBxiArLFC/iPC8B1xoId04CoMDpJI7eG+COrRDIZp8gjU93oCzulz7Q/doYp3eg6hBfj1d6bv3kFC3tPgO7iOXjPWGPsHzHX6tSurdfQLYwZGxGjHoxlxhmRIuw50S70JzobgnfRjEV+698e4KxpI/QHY/G8+dzqM9oO3RBdV8cGEnxEXCwOgKdYs1ZN3cY3x3j9GzZs1LUFNOpDDz2kCpsQIo1JtAgFxYJniBQNhKeGIqfhmTdlCo4wO4aI0DiL1egMi7nz5snSJUtU4CBgdCTGig4i8sA9CQfilSKECCtpBGC5KESmF+h0FCnPhNLFCDZt2lQFgnIRHBbWQvwcwgfWxTBfT38xsOhfBidGsEmTJuo10P5VDFkhAkl/MGVKZIfFxkQDGVgs2jNMQX4ePVoHIp4bfQZ5QB6ROQsMPguguQ+hX8qDJGGYId54K8OHDVdizBQWyopF2ZAzlJ0FcsUUEM+OfG3bvk0/R+ExtUY5+/bvU2VBJOT2Brfrc1BnyAwEBSXE1I9dH4ASZb4eIKf8FsNNdAqSwDjgXoSiKX/turWqTJB9wvoYfKYkkWu+R/0mG0VLuJ3fQETGjh2rbUOom/ujGJFrFpLa8DdjkHbAALA+iWdifDJmUOoQWMYwxNCOURvJ4nmI8NAe1JF0B4TbIbJJTD34F/LEc7Luh+dm3SCkjfdTp02Vexo1kuLFiksvcx++T1uycPxPXrHp22nmuW73LdKk72gzZIT6PvDAAzrVSf0wOihj1lKg2Fm0TeifZyV0j15iWsLB4UpgCQh2A1sBwWZKFj0D5s6dqy/GD1PUkGycBf5Fv1m9hawhy4xzZJGpb6biWJOEw0D0+dFHH9VysF18l/KwS9g29ICdicGe+i8HCARjjA5YtHCh3hPygj39aeRI2WWeizHOuiHGPbqMyD1khjVV1JloLuOX37LUoVbtWqqjf1/+u44/nFd0Fo4l7YF+4Hfcg6AG+hqHac2aNVo2ddhsykHXEDC52bQTkd7YwnWR+RpljvIhnIaRig4eMabrMcFW52q/Hy7Aaq3Sd7i+gHxBaAIBcsSUHX3K7ql/AmFq1rfZ6FCouNZyDFFEhgNtJ5QzzgDkFM8wOiAxkB42KEAMo4NxTxsHMnZ4TtrFv224T0yKEo+ZaU+UMdN1FtHLxejg6DDNDyn11zkQpuHDh2s02G4WcXAIJ5BT5NpGaq+EK8n5tURMz8A1xkv06T1/UD++B3G73nFdWHKUMaz3SkbgStdjAt8N5PvhQiCK3eHaIlBjD5AhvB+8uejTstEBOSByyY6NcJAicK3lGIUcTDux0JO1QpAGyIU/ThjvjvU4fB4TKQIo2EDHDt+P3jZXMhYoaqbhUMpMx1tEL5eIMN4okSde9v42EgypcqTIIbaAnP4TKQJxTYpATM/Atb8jRYDxdiOQInBdRIwcHK5XMKVCNIVw75WICgaVYWTXyP3XAOlgjRWk0J9c4QUzNcUUVlyDfsRbtdOA0UEInzqg3Iku2XrQt/Q7jpuDg8N/A44YOTg4ODg4ODj44IjRdQy79ooQJF45L7xXwpZ4tP4RDBbAsWj8SlGNQECEhEW7lIN44Pmz2M7eO5xlWeDRU1b0uvpP8fA5CQdZzMjCvFBBGUyB2XpSbxbX8zcLn4mA+C/IDhb0Ie1JNIJ7Uy7/8p662RA00RUWipPQ9EqRjUBAmZTN/fmXtmSRJlEQ6sdCZuobLFggyYv7+/eZrZf/VBWfsWiehG+hwranvT/ySUTH/z0L3q80fRcM6C9kFFAOZXCN+jI9YKcIuMazWdDmMU09XC0oh3L9xyKRSdoT/QD4m361chQM6Efu518O4H30vgQHDx2UbVu3qQyFMn3sP+4Zf9zLf0qJhf8sSmbDAnUMBtzfv+/8ZdW/7+g3vmtBe16LqR/amk0lgaR+YGE2C5KvxfP9HWgvptDRybQpfefff7Qp/Yoc8bmVp38CupD+uZyr6Rrjz9LucF2BXVDscmJ3zzPPPKPrNdgxxLoOdvAAdi2wA++uu+76y3bqYMEuAHLLsBOKhHnsUEJIKYuEZWQv9Vcg4QC7BNm1VKRoEXnt9de0fMpiWzxg1xD1ZGs1Bs9/IW2wwMCyNoZFt5RD0kN2unHQKDspeJFKP1SgHFiczA6rW+veqs/OeiQWK1NPwCJldoSx64nEk6xrCRVsgecYC3Z7kDme3Wrs/GQnJn1InWn3YMGuG/IJQeQoh7Yk9QXtxu4cf3AcADvGUI6hAiPOrhfSMbC1mS3/1pAD1oaxgy2cMor8Q8pJdkm/ffDhB7pTkXa89957fd+KWoQPmWYXLcaco0hCAbJDDhramF25tl39y+EFEQwFOAjkYKOchnc3lJ69euraMN5PnTbN9y3RnZTI0ltvvqXjJ1iyYsFuThKmMgY52gmZAjt27tRdT+Swow2uxpheCVZ3cYQF5XD0Dpsk6DcySQN2hbFTjB1gjBf6jhQgsQ3aHR3PGrdAwO5Kktra9oorIIfsxsVG4PQwzv0BaWIsYsPY/UnWb8YtO4SRnysBueB3EMA4gVEeDtcxunfvjnb3WrRooe83bdrkffDBB97UqVO9c+fP6d/Zs2f3UqZM6V2I4XC+YLF//34tt1KlSp4RTj3Azxg9L2vWrF7q1Kk9Q8J83wwfWrVqpWV+9dVXemjpli1bvLp16+pnhljoAaUcrGi8JD0I1BBH/SwUGAWjZVauXFkPHB0/frwezGiUlV43CtLbuHGj79vBwxAuzyh3r0yZMp7xgLUsQ468Tp066efGKHhjxozxGjdurOXa66HiySefvNymgEObR40a5T366KN6vX379no9WBhSovd5+umn9b0hY3pgK9ctOOQxIiLCM0YnxgMkgwHymDZtWs+Qgz8dcPnrr796qVKl8gxZCFtZ/mjSpInW1xAzzxg1zxhczxhX/ezs2bNexYoVVWYNYfIeeuihsBwsTftRpnF+9OBbZIf716hRQ681bdpUx2uoQM4pxxB0HVvG6HrGkGldAbrHEAuVHUNw9Vo4QD0YG8OHD9f327dv94zx9AzZ/sthwaHAODxaP3QJfbd69WrPOCL6GYdFG8LrGadTD21OkSKFt2zZMv0stoB8vvjSS96rr776J1mNjIzUfvYH73lm/3ZHH9MX4ZCxYID+Mk6QHv46b948L3PmzFqX6LB63R4Yz0HxvDfER98D6h+93oaYa39w/VrDRYyucxB6BIRMWSA6c9YsPTQQbzFRwkS6pZyjHv6OfQcDG5bnX8LPhLjteT/hLssCzw4wbcduILxTdhQB8gURnSAxKBEJYL8fCmhf6sdUDNEMvB6iREQdyKpsBmxYyrFTEbQnoWY8Vs4vIwIBiATivVY39SRETtnhgA1r26k5zmUiIy/RB/Ij1ahZQ68HCyuflEMkh8NuyS1FpmpAH7LVPRxt6A8rn5Rv68iUC5Ekygp3eRb2vkxnUVfkgwzdgN2JLOAm0kreGSJK4ZjqsG1MmdSbvG6bfNNLRFXDtWPOyijlMf1BxIQoH1E50KlzZ/XkkR3yShn7oddDhe1Dxj1gnFNOpcqVZP2G9XotHPDvO7uTlHEOiMKTh4sXmdTJ/0PkKDZB9K9Xz54abaXtmUYnrQX9ia5luhsQ5eU6x95wsCzTboAIOrnWiHRfazDWSGNCbi/6j+fnhbyTW4k+JJIE/G0JIArIFBmHQTPzQT4jjgHjQFmiYCzVAOgR/iYSfa3hiNENAkKtCGGkIUf23CuAMQ/H9MTVAEXIK7aMjgVJA1ECTOlhZACEjDxXrIMgCScJzmzitHCBenFPFDTkDwXEVKLNOhsqUAxMCaJ4SYhGcjf6ErATiqnSz9q31zVANuN2uGCNGHVDIZG8kDA8WbHDAUL7KHhC5yRgY0qCvkI5QhIgSrSpNb7hBP3GGCBhJESBZJ2xWRaA2DL1xHsICqBdaQeUPM/BKzacCKaFVq1cqeVjPDCQyFW4QLoCDBTkh76EQDOlMcgYKPqVY3cYFxwgGm7QXiQJpVySIbJEwDpH4QLT2kxlIzO27yB7dvckpInp5lCm764GZK/GYbGkFiLNqfNM9zFuIB44wxzDAQnn+Zi64iBtgM4gmSn3gaBfSyDnTMPb3GVWv0CMSMrIekJLLKMTaKZ+IadM7fMiDxq6j3Wj/DtkyBD9Hkkf+S3j6lrDEaMbBAwA5vutkOHR/VvBIkRIg62rv3EhIoBxJztsbIK1P0RuwlkO9SEyRbTPrs1AoXEd0mSzaOPJWm8rNlClShWNAhDhIfliOIDShsT6kxEUHEoSsskiWuqFpxlu0HaUhceNsbBl2SNOwgkrk5BlDJp9j5FlLdB+Q2iRHYwbR5cQLQsXKItykMkDBw9cTqrJMUisfwoX8OaRURupApBcomGs72tvyDuLmTHQZDcOF5AdnBHIAOtQ+vfrLxfOX1ASGs5yMMCQPdt3/qQC+WS9FuMwNkF7Iq8QQJsKgmg16wshHFxDjgEZ/CEJOB30vyXBRPEhqkRVkPdrCZuzzEZrAeOQkwbQKTiuNrL/d6A+rDEjwzxrItFJ9L8F45mF2Hbx/LWCI0Y3CIiYoHgZ1CciT8icOXPU24/t6A33j+0yogMDy9QSyRUZ8PagWDzlkaNGSZeuXXWgoFhiA2SwhoBBTo6fOC5bt231fRIaUAJ4VByKirIDLM4l9AxQfpRJ/VEQVnGHGxh0vH2mC8K1eBPy07p1azXUkScjZd78edpveP0QPqZGiIKFYzG7hW0fDCoheY5LoCy8WQzpV199pZ/HBli4/tJLLymJoCwiukQ0MxqCSAQHssJYDafBsuOQSEaG9Bk0Qjf4+8GSv0B+NR7hAkaZha/WsLExgOgtwElhJyNHw2DAwu2gUTYGFuKA4ad9iaCGc5Exck/9OOIG8uG/QJ7zCbkOMYxN0JfIrXWMAM4vmz+OHD2iz8DntAW6n80YRFmAdT74nW3/2NIVV4KVRf9yuQZJY/wRkbtS1JvpdWQHBxhiRwSJKCXHmCBbtn7Ats+1rp8jRtc5bLSE6SOA4u/wWQf1ahIk/GObtJFKCafo6D0NGHj2GRBSf+8q3LDCT10JcTNwWn3USs/Twlsl5LrM1PuTjz9Wj87/wNBgYdvPeil4Y+wSwQNnPc49je6RdWtD3yFG3VAcKHx22bC+iBA48+ecccS0CwYI4sT0GsrbKp9QYBUnhg1PlOMxICsYHcL4lBsKrGzQF9SRKZYe3/aQiRMm6g41jtdgTQzbuvHSWV8QDtBntA/lUzemfshsPXnyZN0Bg5FBXsINfxkl6kZU5aPWH6l8QjbZPYriz5gpo+TPn1+naEKFHXO0MXVGZpo2baoRsYjUEVKxQkWNJocKWzecDgwy69/Y8Td8xHCNCNK3TK8xVvgOZYZjbRN9iCNAG1JOqdKl9DwwjCWApIRjvY+tH2McYkfkGf1izxXj8/HjxuvutHBP00cH45w6UTY6gXGK04BOSJ8uvT4j7YHzBwH3j5rY6Bm/IXIDAbdRp2sFeyC01S8AOWXnHw4SOobpen9Ym8JaIogua2UhRm+88YaSKSL01JH6WSJMG9BP17p+jhhdx0BYWKCHEE6fPl2VLus1WEsBywZ9+/XTgYXR6fI3LD0QMO/LvD6eCh7vyJE/qTIcYBQyAwHlRVk21BsOYDwhBhg0juBg8SHkYdzYcVK2TFnp2bOnRh1QKEQGeA4UQijAE2XxIvXE0PUfMEAXnFIOIWIiHngwdh1QsKCdaE/6iHZkHQFRBerIgMc7pO4YctaMYIDY1hoqOCQV40m5Yw1BWfb77xoNo1zKZxF2KOVAAFj7QPuxUJRIGCF0FB/XaDfWFrHmB3nFwIVjvRZGgnU8EDuIAmtSiCww/UJ5lMUrVPnwB0qdKCLkkkgmh2CynRxDsGzpMp0647k4FJMx+tqrr0mbNq3DIjussaBMvHGm5zAeLLjlAE/Wn7zw4guX1+IFC9YQYZjpN6Y0iNjSl6yHy5Y16qBTptAwUvxL9IKNEKFs14eIID8YP8bAcNO+RE8HDhiofcu2esilXdweLCiHNsQBoR7oT9t38+fN174DGGOMcv3b6uv72AakGZmB3KBvkFnIAdN5TFmikyAbRCfpcyIpjB8bQUOX0G+kMPGf9rwWgNThgKxes0bfo1eoA+MDwkxqEmT0U9N3OJnUCyeJdbL0Mf2BAwqoK8+PrUGOIcXIBPW3BP1awyV4dHC4DoDhRTFgaPzn7cMNhjuKFYIZ24tL/4sg+gFppx/DEfG7EiiDspImM+WY/64liNARcYxNMB5w+JBR/6mV2ARjg8jVtRoX9B/rcRiPkLVA5QVSDInFaYzt/ogJOAc4k+MN4cExCTfIo4UTjDPgIkYODv9BoPzxwmOTFAGUL+U4UhQ7wPPFSMUmKQLIiZZzjUkRuBZGmPFAOdeKFAH67FqOC2SFbe1ExZiyDAQkpCSawrrTuCBF4NnnnpUH7r8/VnYnkq6FhfisubzWpAiEHDEizwWVwAN1cHBwcHBwuDpAjogYsXyBqd+rMceQRabQiNLw8l/ncy3BczDVx3R9uFKaAO5rl4QwBUdkLVAw9ciyBKaEg0HIxIhwp11U5eDg4ODg4HD1gAgQrQpkYwvRIkx3XNtenptnCYa8/B2CaRN/0DZEVYONwLs1Rg4ODg4ODg4OPsQKMSK0x2p72CxsknCfP3NjRT1blima3QdxMYfo4ODg4ODg4BAdsbKyDWJEtlK2HzL3aFOYA859YXse21jZtmzzFTg4ODg4ODg4xDVihRiRAZZD+TQLbmSknn1lk1ORvZiFY5AnkhWSN+PvwO8gT8w12vlUok0szoo+v8p25+hEi3wIvPzBlkx+H31elO85oubg4ODg4PDfRaytMSI5HjkI2MrHtjuOsCCTMdEjkhWS7IkzhRo2bKhJ5ngMVpGTs4DEVSRwI1kUByWS4BAiRYSJhIdk1SQzKASpQ4cO+jlnEpGGH7JD5lLOE+JenJbM92677TZ54IEH9JA6EieSZArSxqnFnMdCkji2QFIOib84vNDBwcHBwcHhv4VYSxJh1w+RDZfIDhmFyY4JiWELHuA664843gJSw1ojzoQheywRIhJIkZmYLJr8jmgORxqQGp9rZM+EJJE2HXLFIXzcm0ypnJ3UtGlTzbZL2nrSk3ONrK1sLyTDJgdPQoZ4Lo4U4CA7snaSkdOmo3dwcHBwcHD47yBWs2ex4Jp03hwhQaImyBFp7NniD2ywioRzbM2DGNl8SJAmpuKI4EBaIDeQoxYtWuhUHYSI77Ctj6MPiABxCOcbb74hv4z5RTOCEhEi6kQUiTNZyBnxyisvK2Ej9xJl8ixM75GWHcLEdBrkiiiTg4ODg4ODw38LsUqMICSQm3vuuUfX9XBODany7dogiIkFf/u/5/uQKqbEiBRxZg/EBbLDv5Akm6WU+0FkFixcKHnz5JVaNWtdzn/AIm+iRX369JGcOXPK9u07NBoEUbKZVS1B4+DA5s2ba/SKaT8HBwcHBweH/xZijRhxABwnB7Ndn3VEkA5OCQaQHmBPRydSBLlh/dD27duVrIC+ffvqIm7OTOEwRaJCn332maYC4P4szCbaQ8ZQ7t+nd2/p16+fTpdlzJRJiRO747gv026cvM17DqljATi/Z+qOg0s5aLPrV131ME/KdVNpDg4ODg4O/z0k+MjA93fYAHEZOnSorF69WiMzVapUUfLCydcsyl68ePHlVOZElLjOOiOIUkSaCIlIHaFb/RMmSihDfxyqEZ0777xT7r33Xj3RG7LFPSFYLJx+7LHH9P5Mm7GOiIXWDRo00AgVJyoz1cYOuPr16+vCa6buSpYsKUmSJpG0adLKc889p8Rq/br18ttvvymRq1Spkq82Dg4ODg4ODv8V3BCZr+1aomBA9fyn6BwcHBwcHBwcrgR3JIiDg4ODg4ODgw+xuvjawcHBwcHBweFGgiNGDg4ODg4ODg4+xCoxYms9C6rtLjR/sL3eZq92cHBwcHBwcLgeEKvEiCSMZJPu2rWr70oUNm7aJDfffLMe8bF3717f1WsPtuu/+eabmhcpGED4bL4kBwcHBwcHhxsfsUqMOKKD4zrIOWRBFKnZu+/KJkOOSNrINvu4AnmO2KZPosdAQZ6jn3/++fLhuA4ODg4ODg43PmIlj5EFyRLXrFmjR3CQGwj8+OOP0rdfXylTuoy8+uqrei0mQDggHr/++qsmZGS7Pkd1MDXHPebPny8RERGaC6lbt256HAjvOayWjXbkI+I7/M0Bs+RWIus2h9qS34hs2hxkW65cOU3uSPJHkkXyrNyPI0g4yoRUAWPHjtXnWLFihT4Hh9y++OKLen9yK3FECb/n0FoIH/eOS8Ln4ODg4ODgEBxiNWLE6fovvPiCZpeGLMybN0+JUqmSpTSj9ZXA+WVkuOZUfojLhx9+qOuReL3wwgtKZPbt26cn8HN8xzfffKPvyY4Nz+N3ZNMmA/ZXX32lBIjs2ZAXSM+0adP0qBDObps0aZJGjbg2cOBAPfF/zJgxSnoAx4lwrhoJIdu1ayebN2/W7zMFyCn8kKeePXvqvUkq+fHHHyuRc3BwcHBwcLjxEGvE6OTJk7r+JnOmzHL69Gldx8Np9lWrVtVjP2wEKSZwuv6SJUvk+eefV+KRPXt2nZZ75ZVXNCM2BITDaIna2BP1ifCQ4ZpIE9m2OQwWEvPII4/o50SMit50kx7/QbZtvgdZu/vuu7VMno+s2rfffrtGgChn+vTpsnDhQo1sEZHiWSBI/I66cZYbmbZ79+6tB93yHablbrnlFr2ng4ODg4ODw42FWCNGTKOVL19eiQhHeBB5efLJJ5WwQDw4BDYmMPXFeWf333+/nrT/5ZdfSunSpZXYcGzI008/rd8rXry4vpjegpAwxcWaJcokosT02aOPPqpHh/AsL7/8spw1BI0T9lkQTjTLEiCiWER5Pv/8c41W8bw834gRI/QoESJEvXr1khw5cijpYmrtJkOy+M7XX3+tR5BQT6bp+K6bRnNwcHBwcLgxEWvEaMqUKUpYWM8DGSpWrJhGW1irw440CARreti55o9Lhhhx5hnrhYgwsW4HosE0HFEapsv4l2kvprOY8sqfP7+SGNYHHTl8WDZv2aLTX0x1cTYbi6w5D23ipEk6xcY6JNYgEWWifP7mOWvWrKkRIMgZ57xBljJkzCATJkzQ6TLWKBGFgmhR1kJTxpatW/VZuQaJgzAR7XJwcHBwcHC48RB2YgSpYCoMIsO6HfDGG2/Ia6+9puQCMkT0h8NdWZfDGiEWUFskiB9fp6JYzwMBKlOmjE5dcYjsPffco/dhiz3TWryYJqMsIkX58uWTzFmySOSJE1K4cGG59dZblcg0adJE750oUSJ96QGyhiyxW65IkSL6PA8++KASH/7mM6JR9erVk86dOusUW/Xq1TTCBJliavDY8WOSL29eqVe3rgwaNEiWLVum5bEOCkLo4ODg4ODgcOPhuj0rjceK6fBXdolx/Xo6GPZKz+rg4ODg4OBwY8EdIuvg4ODg4ODg4EOsbtd3cHBwcHBwcLiR4IiRg4ODg4ODg4MPsUqMOCCWF4uoo4O1QuQ6iumAWcCCbHINsdCZLfShgnK2bdsWlnvFFWhHFq37g2u0Ee1Mm4YK7sWi9OgzrCyaj564kkXx5HqiXUMFGcOp24ULfyzEB+wSZJdidJBigeNmQsWFCxc0DQRJSP3rzPUZM2boc4UDyDNJSP2BLC5evFgX7McE8n3xebBAHijDjkP/scamBFJdhENmAEfkTJ06VcuJDsqgLMoMFewWZScpcmpB29o68opeJ3bFskuVrPmhAp2ETFhZ8W9f/rZlUxZlHj78x3FIwYJ7cy82gcQEduqSfiSUVRGM+Zhkcf/+/dqvlGFBOYwX+oJxEiqQc3YX07bRQZsy1sMB6sD9/EHb7tmzx/fuz+B5yI3HhpxrAdrBtnNMY9f2LzrfXvffuBSXYIxhB6Lbep6bzVEHDhzwXfkD1C8mmeM+6P1z5+LOVscqMWKwsgWe/D7+IDM1O7jIO8TfMQFhZTcZCRpbtmwp9evX18EYDBjAJHIkK/aViNiNALKBs8PPwu6mI/0BO+j2H9jv+yQ4kGaA+33xxRd/UrLkoCI9QpcuXXTHIYAokXCT/mvbtq3uzAsWZC4nxxVJP9u2bafXGGgk8CSJJrsUKQNlwXOxY7Fnr56a2Zz8UsGC+73brJn8NHKk1pH6AYw8+bIwgByAzPOFAlJXUAdya1lQv5dfeVlGjx6tZUHOLFCOtC27OFNHpPZdDRy0DTnAyMVF7q5nn31WZaZ///6a44tdnmSVDxVDhw6V1q1b65ilH/3HKakwuEaKDNJahAKyyg8ePFhJFuk3OHdx7dq1KjeMgerVq+uOU5sChP4lzxhjBicsmDMRowN5eP/993WzBfVEr1A2eq5s2bLanzwTbf3YY49J48b3Ba23AMcPsRP31wkT5K233pIffvjB90kUIWRcoB/J+RbsBhD65q677tLjjdDLpB4BGLQWLVqoPPIvThCg/tSbRLvUMxTjzJigXzltgFMNopMUymoV4qlV6Kj//e9/mqgXObDAKFNv9F109OnTR2XsROQJSZ4iue9q7AGdQ/szXgD61H/sNm3a9DKB6Nixo54IwWfI2/WATp07qy7xlwVS82C/d+/erTu6/XEl/cdpFOgL9P7773/wFyJ7zWAMTazBeCFYV6/5++/7rnieqaj3+OOPe6lSpfJWrVrlu/pXmAb2zMDzjBB4RgF6RoA9IxyeUXa+b1w9DBnyjCHwWrRs4bty48EoK69o0aKeUb6+K55nBo9nDKhnDIW3cuVKz7B13yfB4dTp054hX9o/9l6//PKLZwahd/ToUW/y5Mle1apVPaNoPGMIPaOo9TvTpk/3jCAHVf6+ffs8Q5xVLvr27evlzJlT+5uyPv/8c/2bfzNnzuytX7/eM+TNM4bIW79hg2c8Sa9UqVKeIb6+uwWGYcOGeYZQ6t+G6Hn58+f3jFHzunfv7j308MN6fcaMGVo3o5T0faCgTYxX7dWpU8czBsB31fM6deqk8g2++uqry38bheFVq1bNM8pa3wcL2vXTTz/1jLfv7dixwzMG1Hv6mae9Y8eOaXmMI/q6SJEiQY0pC8YWbTjkxx/1/TvvvOMZ4qp/z5s/T2Vn0qRJ+j4UGMPhGdKj/4I77rjDmzZtmmeUr9bDkFh9Tzsag6ffafVRK6/h3XdrncMBQwy8ggULenXr1tX3hqRpPxri4y1btkz7jbE4btw4b/bs2SozxYoVU/kNFl9++aVnHET9mzFRuHBhb8uWLfreGHrvUfPZqVOn9H0wYNwZh8CbMGGCZwiQV6lSJR3/Z86c8QzpuSyHhmTrczD2mzdvruNyyJAhXpo0abyJEyfqdwIFcklbzp8/X8fJ/fff733wwQe+Tz1v/PjxXo4cOTzjFPuuBA9DdLwqVapoHS2ow/PPP+89/sQTvitRaNOmjZYZ7JgPFNT97bffVluJ7Jw2epixS39v375dZazJo1EygP577PHHtM3QWeGS7VBgCLSXPXt21ZM8OzCE16tQvrzq05gQXf8xnmlv4wxqnUaOHKn33Lhxo37nWiNWI0ZEFYwxl0N+YVjyFpEwETZMMkRAEkYiEhwOa5kjUR68B1g0UR68FmM0L3tFhF5JAMlvLUtlKobjOTh81no3AK8L1lq+fAXflRsLeDwkuiQbuGXeMGnOfzNKSZk5STDJ/h0KkiVNqlnDjVxcbuc5pn1z5Y46oJdEmkQc8IDJ1UQ7k6tq6pQp6tUEUz7l4Znh8ZI0kygheaTIS4UnQUJOvDrkCG/PEDXJljWrFCxQQDOPk3sKzzYYkHuKMgHHxKRMmVKnz2jLhQsWaHQMDw45NURevxcoaBMyqadNm1b/tSAhKZEGQOZ0ogzIvFEYWk88wvXrN2jUIxjQX7QrZxLSTshLzRo1tb5kgad/aXvygwUbaQBkkqdu3xkPl4ijMXaabd4oe3mvWTNN7JrKlBlTKD0QUB/kokOHDtpH5CNDH1AWec3IYUY9OAORqNGcuXNkQP8BWj+O+wklqgGYBkX2jFHS/GmA44M4tojs+UydUW6uXLk0ksLRRzwjRxYRTQoWyKgtj7rSj+hVQxpUlz740EMqO8FGwpEvosREnEm6S9SGKSTkFvkgQsexSLQ59WCMEKFiXPI7+pcpnWCATiYKgkxSHmOeaBXTc0RrOeOSyKb/uAkWyGj0iCF1oM5G4fmuREU4+xu5ufvuhjqlY8iT75PYAxFXdC56gClidBqROKLMnLbAtVo1a+l3iRiOHTNW+4V2s/orrkCSZZ7/iSceV13AiylBIsh3N2qkkaGYxn50/YfuY6qdyB51Qs6wd8hgXCBWiRHkhCkBDCkNxCBf9vsyVSD2ENnvv/9ej9hIly6dGnrmFsGqVat03hvji4FE4TBoAeHjMWPHqGDTAXQE4WAOjGWgEmocb8iCBQMQxVXeGJsbDSiuYcOGifFGlUxaBY8i6d69u4YqaTf/KbZQwAAF1lgeNkQouU+hYJgYtLQnShHCAGlBiYVaPsSPqVWUAIoQQ2i8Uf2MtU0YPOOt699pjawAvsdAjPQLjwcCZA6ZtAOXuqPkMWzGk9WpJuQKGaaccIK1GxwjA6grRIK1FBxqTL0xRI888rAqnWBAX6nSN0B5cW9C7wAjyrhjajkUUgT4vfHyVbHVqVNHatWqpeUwjb527TqtC6QJgsJ3ggWKkilUHJ9nnnlGnnrqKSXqEFbbNxg1CBLvfxrxkz4bmfDfeecdnY4K1oADpjUhDxyAzRhkXFI3a2wh5xAka6gYE0wF47jx7MGC+7G+CLKCfPKifj98/4PKzPYd2+Xtt9/WsRMMGM8kyQUQkuUrlkvdenWV1EGg06ZLq9Nr1NMaLX4D0NUQjipVq+j7QEHfoeOtzqc+9BmOATqPKVL6k+vhgNVt/oh+jfF34sRx4wAelebNm2vb4ljHFrCNOPmQe3/ybskw7UOwgLEFcBw5ugpnGOJwpfVR1wqc9kDwIk+evNpPyCZEmunuCxcv6HQ+z4yN9kd0/YdOwkYDAifYE5tsOS4Qa8SIimLkYLwIFlEPzkCrXKmyGlY6Fc+nb9++quTIcI1nSyZqAKl6+OGHteMRDpgyDcmcJML7ysuvqAfzjBmsRBtQftwHFsrgYk2SxS9jftEzzrj/jYZJkydrVAbCt27dWjlhPBgrZJBLvDeibQhTbCwSpC3tgEXweUFIEGKiRgxYPLsBAwbod4IFBpU1GdzHPwJE3+NdMO8MEebIGKsoraEI1rRTHnVg7cFLL72kHiIHFtsFpXhtkHbWG8QGbD0wsrQp5RMZZb0T/Yp8B2vw/EFEAWNmM7IzXiB+Dz30kLa3f3Q1GCB3REjIFs+zQ16pB2WyfoS1YPQT5CIUEF2AiBM1hdShXC1wvFDG1oAQJaKO7777rq6PwdCisIMBpB0CTSSRtkQmIT4WvGdMEh2yoK2RWQwFUcCYjPLVAHLC+MNQPPvcs0ruIFqHDh/SdScvvvCi8dafUKLLOpVQwEkEhw4ekiefeFLfQ6izZsmqfcs44HMLngOjxzOkTxdFwAMFUWDGGI5smzZtdN0gbcx6I/qSNqSd0TV7zLPY8RJboJ1Zd8S4YG0TMkZkBucoNnDK2Md2ZmyUKlVK7SP15Dgr/3VQbDBBhiCIgO/S5kTWsLH+fXKtge1mTEKskX/+hughN6z3w0Yz/ogEsYA/Ovz1nz+w4dSRoAi2Pi4Qa8SIjiPCQSgM7wOFCRmiM1HMnE3Gwju+QyRg6rSpaoQhAChVWDIDkkFPGJsGB0xvYETpDLyXBx94QKfg8B4rVaqkkaTo0aG5c+besCfe7zACR31QkAMHDpKNGzZcnm6xwFhg0AmzhwtWkeMRWqKA4uAFQWVBMkQF4cUADBo0KCTvhegNZIApJLvzjvpgTAm12mhHfqMgbHibZ+HZ+G0w4HgXlDARN0gz5BnvhwW74NsePeTzL77QSKS/EQ4HkH8UCeDeeOQoP8aK9ZKQ8/iGMF2MpjgCBX1EvfxB3Ym84pCE0m+MZ9oPJ4YoLcYMLxLjTV0gR8gJ/ReK5w1ppBw2cnxvnCRkYKhfNM0aUOoD/NsRA8wRQXweDIheo5MYgxgDFnd369bN96nobjHqhi6zwJhB0nhmiCLELRigGzF+OH5p06TVBd60JcaEdgWUQ3Qw2PoBCAHlMFWJfub+OAQYKabtiJbRrwDdACli6iu6XAUKojIQT+qUKmUqady4se52hSgQccMpIqLUzYzJcOq3mGAj1fQdQB8QFccZjQ0cPHBAPNPOOLZEppCTnkbnEGW0oG1iamPsH/bU3w5caxDFpG8YF4xFZAjdyZhn7PGiDbHLVtdZRNd/ODs2goRdJ4KGY0PwJC4Qq8QIhYTBxqOkI5kjZ4cO/9IQhL75HIH48IMPVSgxdIT9UTT83obLLTPGGEKgwO/Lf5fpM2bIRqMUMY4oz49afaRK0BpXFCavW+vequ9vNGBw8HYRPKIaDNQOxsNCediz6PgXb92GX0MBbWvbFxCVwgNgEOC9YFBpXxS9ZfyQV/ooGK+Ygc2AAtyTQYWiRw5QmpBijAO7FfAe7rjzDvWM2cLPehbKxDsJFpSHQYFwQ96pO8/Ei1a48447tL6WHAYL2sq/ffBKmW4CrBPDIWAnBmVB9MF2U7+bK1fW8wODReTJSFVeNpKCcWELMs+CYuK5KlSsqJ8FA+5D+zEuUxkyxJoQ/kYZYmCYJgAYOsZ9sED+uC/9UsyMAfrcvz1pQ6Z8LPgbHcD3iVzzLDYaHSjYDYYTx4vIDUa8VatWvk+jptG4NzqNZ2KcUF+A/EIu0G3BAkOB143BhMwCCKCVH8YBY59oSzBYvmKFRmxY4rB1yxYlQETcIL22jZlGs393Np8jQ4xLSBNT+qGAWQUIJxFSprGJFlid9/jjj+v6RZZM2PWVwYBnjx6ZAP4yxPIECOD8+Wwv36+ECMcwb5Dt+k/A9tHWkD9IKPaOCLGd7Th95rTqRuvUI/+0N7ofok70EvIQV6CfWHfHWjdmaxjz6FCmlJF/ZB87TnQR+0Rklcgj71kC4q//bICEdYroWjvtbZfcXGsk+Ih4YRiBoDHdhWcHYyT0hwcC+yeqgIHD02Fg8122SCdKnEg95rNnzupaEq7RqDQwDUbjMnBoOAY/3grKCMXe+N7GukYE1k1j4jkhMAwoBLtFy5ZKjEqXKq2Cd6OBtqO9aB88e5Tu4489pgMGVo1Hcfr0KV3nATkJBUTlevT41ijhLRq94AUxpX0JKTOdgHdAn/IZQo43DQmGwBHtCRT0DWt4UA7UicGGJ8rUAEbATqUxeO5/4H6pcnMVVdhEeiCErB0JplyAQWF6l3oQOreDELljeolwPnVD+WCI/QljIMABoP0gfnj8GDGIEA4BhAyih2GC3CPvGGCM+ratW3UaCOUcLAhho/CJugHKeuCBB3QN09x5c+XZ/z0rFUx7BwsIB2QZI8YUOW0GkcDYZc+RXb7p/o3KCEaAaRMiOcGAMiiLVAOMb/oCnWLX+HTu3FnLtWOg6E1FZeWqlTJ50mQlFEQ1KxuSGQx4ZsYg+ow6YpTuu+8+fQaMFToN752oI0STCCOGHCWPMX/99dcvR68CBeOd6CzrjJiShLwAyBayyTQPnzEdjH4MFMgk0Sj0KVF6SDl1YqxxP3Q2DidRMvQN68Q+MqQQw8c6F8Yha4wYs4ECsssSAKbSMKpMu6DvbHvTtxA0bAHtGywgx5A35JAF+xhpgL7DbtCn6DT6jzqjF6jbooULdYG9/7KMcIJxSR15UW/0GbMk6B8wc8ZM7Ys7jHOGrOHUsN6WPlq2bKmSVZ47rmBtE/1FP0HYWBeGzYCEosOJ+ODMs84IeSUiiT7FuUC2rf5j/SBRYGQMmUNn4ij6OzvXErFyVpr/Lf2NCdd57/8v7NCGLnmPsET/nv0M8B7DSMPj7VtwjQ7iOt/hPiCme/1bEFM7hArbzsC/vfC6E5mBkDDBHztEKJvrVpkFCwY/g556WFmw8O93f0CCCX2HUi7tR9kYr5hkA4Uaat2Av+z5/0378QyUYeUVcJ36YUypYyigfpTn366MOdo7SdIkpj//3N7BgnJ4oeT925JrlEffxtTGgYLntjLvfz+MbEwRBbuYnzaODdCfVnYtuEa59GmoYxM54H6WAPqD63yOfEYfN4HAyiT/Av92pe+QUSuL9rvA/+9gYPUH9w7l+a8G/nW8muenXfksWEIbDsQ0di9euhjVH4mT/ElnXI9gXNDGdgzQ39TJtmlM+s9Gi/hNbMvE38EdIuvg4ODg4ODg4MP1TTkdHBwcHBwcHK4hHDFycHBwcHBwcPAhVokR84d251JM4HPmFGMC1+36hCt952rB71noxeKwGxm0JYuV/cFMKHO1vMI5K8q9WDgXU24UFr+Ge/s6CyBZwBddXligzGLI6GCXTkzXAwXtxoJSdkLytwWyST3DJTO0Z/Qtq8g35V4puy6L7VkwGgrYEcoOouhlc29kyb/OoQB5YAca7RYT6MfoSd5CgZV5+/IH76PXN1ygfiw8tmPtSs/B+jTkJxyZkymTnYXRxz56Dflgg0KoYIMLi5GjPy/tiPxETzfgX+dw6p3/Mtj9669vWORPn/jLFWAxOmMN/XG9gDV+PJd9Vv5lzFOnvwPyxZoif1i54hVXshWrxIjteOTYYeePPxjI7OpgJ1VMhhdgKFmBz4sdHux6YndEoKDD2ElF5l3KDOYe1wvY6ffmW2/53kWd4M02SXZMsXsi1ENk/cGBtWzFZFcUO4EACpptygMHDZRm772nO5vCAXb1sGuB3Rc2PwyLMu3WVbbQ+m8J5m++1+bjNpe3tgcLDj60W07JowQgXcgbW4fJfs1Oq1BAO3HoJjsILTCczd9vLt8N/k6PlbB5ugDGieeijqEoBnYS0YYQP1If2PwotBk709j1QRbdUEGCTza3YqTZKeY/xiAHXGP3UbgWQKMb2L1INnjSECD/7PoD7CakXux2iQ2wI5a+YVEs+YtIu0D5PAc7bdjhyHV0DvlnSCIais5BTshxgwyxM5ccZgC54Dk4Fgn9iMwGC3bP8azIO3LKjiDALlTujWwyNtj9BtgJSp3ZxckWe57PITRAIJCVhb42Zpcgu86wWy+8+MJlwsouQHawMtY4CSKczkYo6NGjh8oIZAa7hIygt7FLPHN04ACz+wydT9DCgt1pjGtkixepYOICsUqM2EpI3gWYowUDGuOq26/vv/9yNt7oYOskjJnt+ygZ/qbRAgXGgMHOdmK8WmvkbzSwlZaTvbf7KVm2YrOzqH279qokM6QPPY8RwHAuWbpEtz9DWNgmjJdMX7I9+I3X35B6devqdvpQs23jZfCif2wWXMgsRp2dOBgECC0ZlIkooRDsCczPP/e8bv8M1ijQptSJ7dRsWeZvIikM5Mb3Nda8SQxgjMOVIiH/BBQFGZFpJ/9EitRh+e/L5ZOPP9Htw6QLAOzkYFywIwNSarcWBwrGGQ4JW2UhEaQBoI1pWww3ZJBxgbENhXzhtUJUIQU4MeTcoRzAdnn6CQVHDi6bwC1UkMGaBHGcio5OYEcL6QAAbY3j9U+eajCgXPLOWGNkc7XxHMgnni96i+gOxBPCCKn2J8SBgi3c3IPt+LwggDgNtPliQ2gol7ZAfoLpR/oPMknfIe/IKA4Y1zl+he3tpD5hGztpVPiczyDCtAX6Oa7Os/o3gTYnV99F3wwJZAHZ5vrPo39WkgpI3QAZpb9Iokq+p7gG8s4YYMwxFrG52bJnU1uB/oLI7d6z2/ftKLDDkbHDkT02gSYOD7aAtA3YOpvaJC4Qq8QIQsTA8Tee5DiiM1HYJO7C86LTbfpv6zkjGAxCEsaRKwEDZc/OAiuWr5Dx48Zrp9jpFwwkOWCIVFmvB9aKJ4RhoKFDyQkTVyCPEIYUT9ieu2SjNwvmL5BNWzapQQh1a7cFpITtoGyZxKihePFMIQwctkpOGdqSaB9TQaGAxJ8QMO5JbiRIEKQAUk2kgXw4eFLkxiDMzDOgiJEfsqryXQxUMCDnDYSb42Z0MBujxpZvokSN7m6k94c8MI0AwQkGKAruST39t59SJnl1kH/KQ46RfRQCCqJBg9vMsx0KmrRwX2SdHFD0EQkxUUQ8S7NmzXQs8Z5IB98NFsgc9WJcM9WD82Fz7bz15lvabyjHcEwpWUDCIALk8kE3kBOKxJiA98hHuMaCBXKCbiG/D30JyA2FEqcfaQPGIPoOmUHn8BwYMf+jQgIFfUjfIffIKDlucBjQlyT/BNyfqRWMSqDACEPEiXzRV9QHYom8Uw79StQP+SRfGNeIWpGLhlx0tAtj1iF44KQwbrBVHBOCLBEpIWkoL3I82eSdyDV9z1jDLqCP4xJMhRH5wRFgGz7OATmtcFqxGzhGOHvnz/112g/dnjoi4rJuJe0EJAo7h/7n3oyluECsEiOyXL/62quq8FGM/Ltw0UIpXqK4GlzAgMfo8jkhWYwysHPneEwkAGRAMuUAvu7WTYb8OESOHD2iUwSEqiFXsFa+h6dtvVYLogN4OjT6DQVjFyGSEDsEjjpYUM/qNarLa6++phGjcAGjSdgc8oVh54UChHjiQQJIEwLNQA4H8K6pD4OLKRc8BcgSwENHaUOckAtr+FAgDKZj0dY/XC0gPnjhyBdkgcgQGYqpo82Jg7EhyVio+WiiY7ch/RyMDEjsSL9CYFCStP84Q/o5RJZ2CRYQS0gd05EQLA5fBZTF1CHHVdg2DhbIBiQFwwwJgCRQHkacNWoRpi5fff21HvUQfY1MsKC9iEwBHCDqgBKOTRB9g4hVqlhJxwWyT7kQMYDeQkat80ZUjigk7QLhDhZVqlTRtsM5gDTbiCr60rYBBB9yG0yUDH1pxzR1wiGtXae2jkMi9Ze8S0qecT4ZK9TPjgXqBkmM7bb/NwM5Gf/reLV9kFR/FwUdDEE6d/6cZMwUNU4Za4wtjgsiGSRkKi6BTJKZGyeLcYEcIhM2AIHuxm5ZXRcd/o4f48nKNFwB3cKYigvEGjFikDHXmC9vPmV+GBjmIcuWKSsHD0R5GXgiNCzpwW1kwmb9JAoAW8QQEnlCkcOaCTcOM6SH+VfmLxsZb5tBzJwm8+Q2SyiD2YKyCUHymyt10PWKWXNma1tgnKkHjJx2hRjg6XXt0lXPp4Fccj0cIEqHwnzuueeUNKD0IWa0qz8xC2UKJjpKliqpMsDZaKx5sGBRKJ46a34gLZSJAgH8zSv4eEfUKdZELol+kbHYv04ofsrH6wk7/OrBWKE/mSrEC4Tk8sJbJBNsKCCyh8KhDW1kDTLJdRQr7W2dkWCBAqMN6Z/uZpwx7UrUi3JaGrLb2nfga6iHyEYHU0roBzzq2ATT/jhWrH2gXqz7oZ8sGHcYOP/IUJasWdRgQGDQccECQoJnjQdNXWlDG+Gz07t8JxxjETlABp9q+pS+5/7ob4gd0202Cm9BdmhkKKbEmg7/DNqXNX41a9TU/kOeDxgH1M6AFCpcWCOP06ZOk/79opaAMNaQ95SGDDOdGsraslDBkgtsO2OfIIZdgG3BjA/LPdBhVmavFkzDYe+R/bhArBEjWK2ddsHIMl+KJ0nHAiqNskb5E3qfO2+uCgdhcRqb+XwMJesjaHS7SJs04yz84/d4L88assMcPqQKRQRx4vs2/T9Ki3UU9957j7LsGw3LfVMshCo59gDlSHTDf5cI3iReH4IZDrCeACOKQMP+6SOIJpEaiAJgEGPMk5u+DQdyZM+haxYwAOyCASgI1jmgCOyRFqwloWwAseBlvYxAgaJHuRDZxHixxoi/AVFIyCZeMvUONzi6xvYhU8144Rgg2pQ2h1RA8iGiwRwiC9kiIkTbUU/GFXW1MsLaPhwVpiQx6sGC52O9CTLIFHaihAlV2TPli8NCdAcdwFj2V5rhAIqXnYlEcmITkAKcOBwvIjboGP+z0jBO9CVk0yJpkqT6fdbAEc0Odi0eEUPK4xmIqDJtQVQHmbdkyO7qsVN8wYD6oY/bf9Ze+w75YfE/Rg/5YVwiTxbUFwIc11M5NzIg2byY6cDx4xigdm3bKuEEEaaPWWbgv6GBqSaiSLPMWINs2MX4cQHGHvqSpQAQdsiMnbnA7qLP2RyE7gkE6BTkMdj1leFArBIjogwwPogKCh9ihMFFkUGW8MiZHmGAffjBh2os+B4RIjwtBh1rB/guigHQaHadzbr165RArTRkAYOJV8piRMJ6eD4oIwweUYEUKVJqJ4bDs7qWIHrDDhvYOaFtDJkNp1sPDiLBtFA4F6pZ5ci6L9oU43Zbg9u0TIgJZI02hzSFApQ6gwBwX/qaNSQAMr19+zZDhKvpbh9kil0aW7dt1f5l6gDyhNcaDPD6qSMggom84cUhRxyGCGFH+XDA48FDB/V7ocDfayLaZyM4GD7IKNcYDyNGDNfr1I8xFMwhsszrM/3JdCPlMg3Nv7bvAGOM8UYZwYL2o71weBi/KHHIM+tPuGZPx6atMbLhBLKIfqHc6LAOWDiAI8IYhDjYQ2SZdrXgOeg3e04b0wc2okRbMJUQ7NpGyKSVGzYn2Gk5nECiWADihewGu7gdfUt9cCZPnTwlAwYMUKOHrNidhKw9wgmx+pMyIb6BGj2HP4D+hHzi2OO8o9vtTtytW7ZqZBL9hs1DvgDygGyj69mVFq6lDMGAqXn0MrM5LAbHCWM3M8/NvxERqY28ZtVgBraagAc6zzpnyDV1iR5LYvyg+wiAxBVi5RBZ1g3BZPGC8TQwfigUPHAEIUXKFBp2hlXaMDONyUBEAGDQrEeAMaJgaSQYKYMQD5SV+gzMHTt2qhCx4r1vn76XBYmBznU8ZKbQMAQoNubrEb4bCbQhUxQoXQgkhoYpLkglRgiPksHCWivaJhyw0RIEmqk0piwB05zskGGnGOWzmwsSGgrwfqkPCwohePfdf58ansHfD9bFuyxA5llYZ0XqB6J+hw8d1u3fkDabriAYEGVjYSuRIubzUVR4P8zjs0aNNT8DBw5UWWzySBMlbcGAqUGiMyweR3lgyGk3ZJhpGoghW10xbBBNdh5xkC/RQZ4lmIgVESd+Rx/ileJA4GkiS0SimDYhwkMkkrVBwQLiRbvQPxAx2hGZgeghs4x3xiPTLexMw5iGC4xvptut0QCQFLbUo1to63CMCTsGcRbsNCEOC4odo0V5EGvkCfKA/oJoMFbRR4wTuxYpUBDZY6wxNugvSCwyil5kDOAwMoYoL5iIEcTtlVdflYnmXsgppGjf/n3ywP0P6HjQ90a/MOaJ3ts1HxhE+tSuFXUIHDgljBGif4whdDmReeQZR4Yd1LQ7fc50PmMNWYSgQ7w5dPs103fh2u0ZKHgWZm2QA/Qoz/Tggw+qTWd2A+eVTUPI7j2N7tHvEPVkXRSOAuspIYVwBmwz9wM4wESD43JRf6yclYa3ikKg4/E4KAJmCEHibxQKipu/GZgIBZ+jVFCc/J73vPg9Ximkid9wEv+xo8f0/jZcz30wOtzHKisUkS3PPgsNzz1vVFAPXrQJ/xLOps0QTtomXKDdYPW0Jy9/UB7l0pbhWFtA3yID9C/9yT3pIytD9J+VGSsn/AbDh6KwXnqw4F7IDvdFQVEOz8K/lA9ob+s5BwPaDJkE3AdZBFyz8u/fzrR/5MlISZE86oTxUICzwP24D21Lfek/ImNcC7X9AP1D9In6RK+LlVHalv4KJ+g3CAt9ZXGltg4XaD/kwo436o4s+j8H8kO9+Q4yHYrsAOpk5d3KKKCebDxIaXSdNSqBgue3epK68S861T6zys/ZqAONU6aIitQD9ANjJthyHf4K+hN5pV0ZT7xoX/qca+DvxlpcgjGB/CCj6G4rS/zLOKAe6BzkCXuFDPOezwHyZuuIPHI9HPYlWLhDZB0cHBwcHBwcfLhxwycODg4ODg4ODmGGI0YODg4ODg4ODj44YuTg4ODg4ODg4EPYd6X5g4VWLLBioVX0RZBcZ+Epn8W0KJMFXCzw43v+ix2DAQvV2JVjF9jeqGARJlt0/TPp+rcT7WgXsIUKu2iX+9J+3JcXC//YKcFn4czvww4stmmyDdXKCgvw2NHAItvoZXFEAbugQn0G2pRdP+TJIheWbT9SPSAzLAoMxwJl2ott7f73YpEuu7gog0W60cFuOHYcBZvkjLrZMciL9kRGGHPsFGNXHDupwiEz5PLh4Et2yETfeWblFtmJaWt9sKA8ZIYyrcwwFrhO24YzfYUFOou6cm/a8UrjhBQl7JBjcWywO9IsqAs5YtjuHD3LNHJKnUNdqMrOX/oo+vOyA48t1sitv4yyNJXdarR7ODd+/JfBeEeGbDuTX4xdwOw29LeRfA+9iCyEurA/XEBHo0PZhID8I5PoT+zTlcY8Mof+Yyz56wx26TLG2LkWZ7LF4uvYwoQJE7xixYp5nTp18l2JwpYtW7wKFSp4L7zwgv4dE4zC82677Tbv9ttv995++22vatWqnjGGvk8Dw/vvv+898cQTXpEiRbx169b5rt54+OCDD7waNWr43nmeUb7eHXfcoW188803e3v27vF9EhqMgvSqV6/uFS9e3CtbtqyXJ08eb9SoUZ4RVu/555/3Bg8e7L3zzjtely5dfL8IDZ07d9bycuXK5b377rt6zRgBr2nTpl7evHk9Y/i0D42B9QxJ9j5s0cJ7/Y039PNQnsEYM5WLL7/80vv888/1foD616lTR58HmZk2bZpeDwaGjHiTJk3yypUrp+VYUL9HH33Ua926tfahMea+Tzxvx44d3jPPPON9+OGHVxwfVwPaJl++fF6ZMmW8QoUKeQ8++KBnDKnXr18/79lnn9U25RrPEgoGDBjgPfXUU96PP/7o3XPPPZ5R5r5PPM+QaC2jV69eWnY4YIyHyh965euvv/Zef/117/jx454hyt4bRi46dOjgNW7c2OvWrZvvF+FDCyN7d999t/69fPlyr2bNmn8aJ8OGDdPr9957r3fTTTd5FStW1PfBgjo98MADXp8+fbTst956y/dJVP/St8hXKJg6dapXvnx5fX7kxN4PuadfGfvI4/jx4/W6MWj6/s4779S/HUIH4xxdM3bcOH0/zvyL/cuTO4/awIOHDup1xu7TTz99eaytXLlSr8clLpnXO0ZvI6eMTUOmvYceekjlCR3au3fvqC/6AVli3PAdxgk6F7Rr107fZ8+e3bvllls8Q771+rVGrBIjDDfc67333vNd8TzjYXn3mwY0HohnvFbf1b/i3PlzKhhdu3ZVA4YQPP7442poAoHx8DzjdXuGzaoSmTJliu+TGws8NwSorDGwFggcRoF2hLSEC999950aMgz0qlWrVPkjoD169FBjDmbPnu01aNBA2zUUcN/+/fvr399//72XO3duLzIyUuuLYYMIde/e3TNek2c8CW/hggVKArds3eoZz8krWbJk0IbHeF1e6dKlvQtmMAOMA3WmvMlTJnvG61EjSD3PnDmj3wkUPD+oV6+e98knn+jfoH379mpYAG1966236ncZM5UqVdJnCAW0a8eOHbWOGFf+htRynftbhVqrVi2vdZvW+ncwYGzy7ChqwFhv27at/j112lQlDDNnztT34cLcuXPVQThw4IDqk9q1a3tLlixRooSzBebMmeMVLlL4TyQtVEAYChcurHoJDBkyxPv2229VZtasWaNkmn8nTpzoLV68WOUHnUPbB4uffvpJDQQ4ZsgfBBsZARjTunXrej///LO+DwbINeRy1qxZ2pbIQ5MmTfTZIT84DABihrGzGDFihMrRnj3hccb+y2Dcv/baa16iRIm8MWPHKrkYOXKkjlvGabZs2dTBAMgC5BvgRH722Wf6d1yCZ8uZM6faaJxXSF2v3r30MxyYEiVKaKDDH82aNVO7jD4iSPLkk096+/fvV0cRB2rhwoVejhw5VDfGBWJ1jRGhWRI8Hjr4R9bgn0b+pOnMyQZrk4WRkZqkjWQYJkQIVq1cpUmeyHRMmI5MrCSNI3xtnlsTm3GwI1mDCWMDEl6RXIqTy0naBwhLcuI1SbE4HfpGzNRKSJWkmSTPsnkrCK9z3ACJEQcOHBjWaQMSKRoSqlM4JLSjD8l8zbQVZ998990g7bNixYuFnFyMe5J4EBCW52gJcl6QAJQkeoRlyfJLvzFl8YtpB54rT+7cWmfaw8pMoCCBHVMR7T/7TBMhksCShIAkPaxerbrem+SRVuaCgZ2mYirC/g1IOGqPsiBJHuFxEkqSHJCpJ2TdJggMBuQKITOtPcOMvDr2WA6mtOyUMp9TTrAg1I0MkICVKRdjrLUcxiTZ7MkezhQechQuMJXMtBJHj5B1l1A9cjTblG8TOpIkL3my5Nqm4QBTSiTgI9EebQtIfMp75BFdRfJYZIqT5zkehL5j7PA+WKC77Blp3w0apP1l68hzBJtR2x/IO+fAMR1HfZjOYYqGaVZ0LLqWZKr2WB6APNsccg6hAf3N9CV6Dh3HtBmHnSPX9AM6yB6PwbSaHWvYBXtKQFwBuxyV+fopo98S6NQZyRoffuhh/RyZQUdElxPONkXHMW5J0orux1ZzBApyjf7nPujCuECsEiMGFMdYrF23ThOUMee4etVqVZY2YypkiBP0aSTOVrHHFaxcsVKVH0dhYDj5DSnQaWCyZ06bPk1y5MwhxvPWdQYYarILc1+y7SI4YLvpOH7HGS58Fsp5QnEBBGP4iOF6iCxtcMEoLYCw2czUZACmjuECRtPOXbO2AYUPOUCAOf/pySebasbm95u/f3ltR6ignzl2gLUZlIXitcabOWcMHcQFZYChB5SNEgk2LT5GjIzBZJ3mHCiyTGMcrEJC1sgCTr3DnciOekI2AfWEPHBWEmMhbdp0OlYgiSjNYMDzWwPOXD4EAQVL3SApZFEGrJGJHy80NcDYYgxCAFDuHC3B/cl0ixGHBKIgWdMUDpANum27tnroL31HlmD6Mpe5jrzStiho6hYuw43jRsZpMpYzJukvSw4A65o4vohrgDVcEFP6058QBwqyHpMxnPGNLFBff4RaP57fyiFkluzXED7GFrqbQ7c5RQAZtQ6MRbja9r8M1hAx7jkZAkL6R6rSqIPUOSZo1+5dl2WIsYbzQfZoXvZM0LgCgQj0VNas2cyYuKi6G/1s16mhd8isH/3wdv+1cqzF46w4//VtOIo4FuiTuECsESMiGnQ0RISFWBgYjjrgXCsWaeGlEOEhusOp9xg9lJs9YmL6jOkatUC5cS8IFEqdowdYLPvaq69J5UqV5VXDPDGOGDUUCGcG0TmcSgxyGm8Oz5IoCOWzePJGAgcKTp40WdsRpUX0bI0hCihmImAcN8CRGXZxZjhBdIEX0T2AV3zyZKT2F4uCIaLhAsqYA1uRB0tqAUYdw4qRYeBcMsrYRlFQzLSDvzIJBNyHBdF4yeeMrKKcMKgWRB0hGAza2ICtB3VA8TEu6MN2xugjz0RLoxvCYLB69Wo12CgjXnhrjKe2bdvqcRaMq1DA2MZzxfHguYlwEIlDyXHmHJFNDCsHAocDtNeSxUvUUENYOX4AHcH4R+dgPDgiB4JE1CNUoDcgdZBm/kUm/aNsHIUA2cTLtSAajjwR0SEKyDMHA6J7yCR1pZ2bm/aNLRARox2J1AKILaSJ6AVjkKi1Q/gQaWwb50EiK8gTY59zPxk7gIjyBx98ICuWr9DxCpABiAbjDRnHJsQVcBbsmYDICrbC3/lBD2DjOYz8SuCMOGy+DZQAHA+uQ8gJmMQFYo0YYdwwqHbHCAeRwiwhJhg4Bpyd3sIDJALB95gywVix6wjBgBwRxqaRAcaK87LwRFH2DQ3hwZjSuIStCetRBn8Dy2AxvNYA3UhAufPMbB6kbgyMfkZoUGAWsGoMHkYhnMDrRSlDNgHn2kREpNHz5xjQQ4cO1QERDtCXDCBkxt4TQk3YGE+dc69AYUOgiT4CPFwGUbDTiHj5RKpQOhCEdYbsMTULUFRMG0I8bRQgnMBgW6UC4cTock4aRMwSpspG/hMbjz6Y0/X9wXRr48aN9W/GA2QLY8/4whlhXAYL+oADI/FsfzLyybj7+uuvlQgxnnkR3cQhYpogHGAnCwYBGUQmiSjSVxykjH7gIE7GPOPC/8T7YAHpoZ5EF9FZlE201oIoJgTG/zBe2hmZ5RR+CC+fBwPICB41hoKI+ozp0zWaGG7gWOFwEoGnz5BBTvJHRhgjnFvFuHdRovDhqCFAjBPGJ+2OHA01bW3JDg4/4wrCb51enAyWVEBUmV5jtiCugF3CHjEuGHfYaM5GA9jsvn37qRN2pV21cASiX0yf+dtlzudjupxzMOMKscYSCCEzP04FMWQwPw4+hAAx2PD0ID8YNRq0xYct1MPEGBJe5DdEkSBNeF1ETgCG0Hr1i35bpIaMMD6/JYoBgeB3MFfYKsaP72Po8MBtqO5GAYMAJQz5I5zOtBaDCKPKQY4AocQAhHOdEeCgUUiRnVaDWLJWBWDomDoIFRgMDg2E1LH9HtKMQaPPICVcy5AhvSrlsePG6gGoe/dETQ2xpZMBFew8O2VgAJC5WsYLu8mQa+QSOYI8FCxYwJDDDWrciBKEAuqH7FoQhbLTWcg206TUrXjxYjJkyA96ffOmTVK9WrWgTte3OHb8mIbqcSYsaDP6FGLBQcTM5YcCCA+R3WSm7TCg3J8xDiFiOh2Ecz0Eckif4Rwgg+UMmbbmGhKLgWf8o7Cjpw4IBkQyicriCKDomZL0j+RBKnDo7Job2pupAMAaDL6PAQwGyKh1eBo2bKhTEnbrNu2A/FoiHSwgWjiuUevaftcoH2OL+1oy+9BDD2m/QvgAn9kpRYfggG3s2LGjyhWOGc79u82a6UGy2C50PO2Pc2zXGF0ea0ZP4uyEQ76DBUELHJMRI0Zo1J2IKUsSeG6itqlTp5QTJ46rnmE8QpaoK8+PzmdJDPWirkRVd+7aqU4qpI+lE+PHj1O5jAsyHiun648ePVqZI52GwrcLQfFAqDSLrLiOEcK71AhSpow60PLkyaMhOqI+hBjxQFEO/JbGQhFy6jANSwMzYAk9Qh7w6iBSRBweeeQR9bYwanhcLFrkGfyZ6Y0AlCDthCEjkoaBgSxBKgmzDh8+TL9n18eECwxAIlR4LDZXEP0xddpUWbY0inQyxw1BCgUYENYWMbggr88+96yUKllKfhz6o3Tv1l09JfoRkvZIk0ekQvkK6tFyMjMy8M4771yOaAUKZA0CTtkYU2SStTAYBsqDfP388y8ql0z9BquEkD8UCPKKPENgMaREIpgaRvFRJsazXLnyMnbMWFm6bKmGplEewRpVMGP6DJULO1ePQWNBLVEA1k4xLWqNXTCgL4h+UQ9IAG1GnzD1i6FF4SOrOCtPP/20fj9U2GgbbYrCzZ07ly72JDKDR73ZeKGdOnW6vLkjVNgxyL/0Cc4BU/O0G6SFKBKEkOg47csUMwaCMZI2XVp5/bXXlSQGA9oNos7iZ/KHsdaQ6DfAyIwzzhHTwEyvWAcmEFAXSB5jj9eoUaNVH1MfnDAIIWOUKB2OCpFpdAP9ynNRZxuddwgMyA86hReyhQ6qZvQpEZZmhiAxTulzdBJ2DhDhRd4IBrAeF91p1xJea/DMyAo2lefBWeFZsd/oVK4RDSMijjwRHcLeE0lF5iDfRMf4DvdB3tCDECicxvHjf5XyFcqbNqnmK/Ha4YY8RNZ6SP5K9krTZHzX39P5N4Guo97hMDYxAaUfk0KHqHI9nCQTUss9r7afAv3+3wHiw31iqx3/DpRNuf71oF9t24daP+Sfe9i+4t5EWlBE4Wg7C8rhFd0426hGTHIUKmg76mPvTZvxPhiCEApiGifhlinK4F7hHHNXg0veJblw/kLYxprD1QMdB/mI3udXGmvXOxibPDd1ut7hTtd3cHBwcHBwcPDh2rofDg4ODg4ODg7XMRwxcnBwcHBwcHDw4ZoQIxZlMV8a22BWkIWC5PrxB2XzYn7TwcHBwcHBweFKuCbEiARs5BmJbbDqnR1bLVq2+NM2UlIHsGWWTK52u7mDg4ODg4ODQ3TEOjEigsP2erZ3slPjSvAnMux6ir4m3Oac8QffYbeGBVtqCxUuJPny5ru8g4LvkF8BQkQGYP/vs+3Uvxz+jqkc4J9Q0cHBwcHBweHfiVjflUbGVvKXQFrI50L+FkBeDBITklyO/Bkk+nv77bc1YSOHz5J7hdwN5CzhkFCuM0VG6nDyeCxdukRzbqSOiJCUKVJo9l7yOsyYOUOKFyuu+T7IhUDeJJK+kSCQfDfkV+BZyPxLfgUy5JLfiOk+8i+R14Z8KCSs4vvkVCCXB5+TR4mIVFxs63ZwcHBwcHCIfcRqxAiSARmCeBCdiZ6+nOSLnCdEsrkJEyZo0j7SgJMBlOSCZMMkCR4ZNsl9wJQYyeNIJAeJWrBwgWTLmk2T4EF4IE4fvP+BJoMkESK/g+Rw1AKfEUXiNWDgQL03xIfMmoO++04PpHz7nbc1GylEiGclwsVxBxAmjht47rnn9BkcHBwcHBwc/p2IVWJERIhMqRzoSgSI6A1nqwB7fgqZrDnagogQmWLJAgwZIhso0Rqm4chszSnCkBfOQVq9ZrUhXIulapWqcvPNlS+fu0a2XSJAkCjS8lMWx4Bw1hZZcJmuY6rsZ0N0WIzNWWBk4V63dq0UKVpE6tSuoxm3+Yx/eQ6eC4IE0eK+27ZHPb+Dg4ODg4PDvw+xRowgIRCjhIkSavp+UttDdDjzyx+kAeeIBDLxcmQC4Ew1CFHXrl3lvCEl/B4Sw/lLTJmdO3tO1wpBVDhgkxdTXcwK2pc9tJG/yRxqM9PyXDZ6xAG2RIA4pbhkiZL6OUdBkGKde1MGBImpM84pIvrV+N6owzgdHBwcHBwc/n2INWLEOUZMVb3X7D2dgrJHsnGdtT0xHRHAOTDARmkgImUNSQIQJSJKTLeRCp0pL6JLpw0hOsnJ1uZ7ECCIEKSHM9MAB9QSOWL6jc8gOZAxFoVzrhNkjPtyxhJncpUoWVKn/Tijy64r4jwXSB1nv3CSt4ODg4ODg8O/E2E/RBZAOFgDBOng1Pd06dLpCdScdM8hjJAT/mULP1EZDrhkCoyTo9k5BnlicXbdunX1UDqICgfNQZiIPHEyOd9nXRFb9JMlSy6ffdZeD+EjSgWpsicPc5gtxId7QKiYkuOU71mzZikR4sBZDrhjLRKLvpmWi2+IFddZ/A2JolzKZ4qNwyNDOdTTwcHBwcHB4fpFrOxK45Z2+z3RG/9Ijv9nECT+5jovwOdsrWfnmP9p5kyN8Z0UKVL4roicPBWVOJJdZwn97uUPpsIgRLz8nwNCRkSItU/+02z6e/O/Sxf/OJwVUsXaJNYj2WsODg4ODg4O/z64Q2QdHBwcHBwcHHyI9QSPDg4ODg4ODg43ChwxcnBwcHBwcHDwIVaJEet4mKnjiA3WAgUCchexK43dY+EEz8JibHarsW4oJmgG7Rkzrvi5g4ODg4ODw78TsUqM2IXGjq9XXnlFevbsKQ888MBVZ45moTRZp9lVFk6weJqEku3bt1eSROqAfv366UJsQMZtdqB169btmhx86+Dg4ODg4HD9IFaJEUdxEPUpWbKk/O9//9OcQC1btpSDBw/6vnFlsCONRIyVKlfyXbl6TJkyRY4cPeJ791dwX5JKsiONY0uIIIH9+/dL79699Sw2/uVcNgcHBwcHB4f/DmKVGG3evFm3uHMkCETHbr+PvqXeH+Q3gtgM+m6QnD13VooWKarXmVIjuSOZqZkGA2yv//333zWyQ+JFoj5k1ua8tXlz58n06dNl7969Op1HXiSiQ0zprV+/XurUqaO5i5iy+/DDD/VvzmHjPckhyVVEUsdff/1VczCRk4k8ScuWLdNn5IBaUgEAnpeyiIbZyJODg4ODg4PDjYdYJUYkUAScM/b111/reWZMq6VPn16vRwcJFiEpEI/Ro0ZLgfwF9HR9iEfnzp2ViHDoLAkgQafOnWTYsGGa5PGFF15QwkWZnM/GlNmXX36pB8ByvVevXvqZPYuNU/0hMS+//LKSLb7PQbYcEwKGDh2qv+E4Er5z/MRxmThxorRt21Zat24tL774ot6HA2whahAlMnwTjXJwcHBwcHC4MRGrxIhITpWqVZXoMHUFSWGdUUxgSuvTTz+VihUrXs5sDUk5ffq0rjXiIFgyUbOgu1KlSrpOaNTIUfLmm29q9KdJkyaaqJEo0osvvSj169fXLNdEh0D58uU1EsTvOFAWckYZZMmGJHHESJYsWeStt97SI0Qo8/7779fPIWepUqbSexMxoqxx48bJpEmTlPS9+uqr+ntO4OeeDg4ODg4ODjcmYo0YEUVhLdErL7+sZAhSA3G4EtgFdvTYUf3eiuUrZP2G9UpYBg0apKSKY0MgIkR/6tWrJz169JB7771XjxvJmzevPPPMM7Jx0yadOiterLjek8Ngec80GlNh/I4jQ+644w79nIXWkB4IEdEiiFWePHm0TDJlMzXG74YMGaIECTLEWqmqhuyRmZsIFFEiwLEhkDH/zNwODg4ODg4ONxZijRhBFNKmTSvFihXzXYkCRGPhwoUaCfIHp+fnyJ5D1wC1bNVSp7Y4ioN1PaVLl9ZDYN9//33JnDmzfpd1Q5AiAPHhfLaNGzbo2iAOgAWce0a0ijPVnnjiCSVJRJAyZcqkESrWDnGSPxEi7kGECXAkCdNxRI84VZ97s4WfnXKPPPKIfof1R4DngaTNnj1bt/dDoBwcHBwcHBxuTISdGEFYWBPEafisCVq8ZInvkyhAQph64qR6f7BzDTLF1BhTXqwFYtqM66xP6t2nt0RERJEerteqVUs+//xzvdfkKZOVJPHZ8uXLL+cfggDx992N7tbPiWKxuJo1SSwEh2xBlCA8/I5F4oDID+ud7rzzTl1nxFlsRLRKlCgh2bNn1+9AuiBBTz391OVddtzbnbDi4ODg4OBw4yJWzkoj0kPkh2kvCAjTUhbkDmKBMmtxEiZM6LsatcMMYsN0FtNRkCp7ij3EA3LCo/J7rvN9S0iYMqMMymVBNe/57oCBAyRb1mx6Qj+AyBAN4v48lz2slu365C1igTeEB0ByuB/PybZ+/rb1seAZiXxBuvib+/rX1cHBwcHBweHGwr/yEFkWTkOySpYqKU0ebqKn5ccEtvqzfgjiw5QaESIHBwcHBweH/y7+lcSI3ENElogUsVbpSmBHGeudChYsqFN2f/ddBwcHBwcHh38//pXEyMHBwcHBwcEhGMRqHiMHBwcHB4eYwDpU1m7GhH1798qe3Xv0b9aTclxTZGTUSQPBgvWn0XcN8z4yMtL37s/g+Ti9gZ3U1wKsvb1WZYUbtBU7vekrfzBzw6kVrOeNDvqeDVD89npDrBIjBJEXnY3wxRSc4jr5gtj1RSPyXX7D9XBsfd+wYYPe798A2ocEk/6gjWyb8Xm4gICTzoA0BdHBgnPSIIQTlEPd2HFogbywMJ7jXqJj5cqVmkYhWFA/jojxbzPKY3ci6SCig8/YUcluyWDA4Kc8f6AYFixYoNnVYwK7J0kjEQook6zs0TOyk7qC/g3HGAO0DZnhYxprVm5jaterAW1PG/kbUfqPafCY7klbUz9/0G+0Q6iGh9+zk9XqMp6Ja7wYF1ae2LzBIdpXcy7klUDfRJcZC8aLf5/S/rTHlYz8PwHiQWqU6M9LO9Kv/uOdurPDl7HiP16vFrQTu5Y5BQHi4Q/6iUS/PwwZYupyQtuzS5cumgvvvvvu193CgQLZ4Z5t2rT5U/ugP15//XXp3r2778ofIDkx6WGWr1h+ua9jE6yJ5YSFhb7xTs4+UsVwCHvjxo31jFHb3+TZu+uuu+TBBx/UZSPXA77/4QdtY395QEbefffdGPUM8kpOQPq1WbNmlz/nsHnyE1JvPnv77bf1+rVGrBIjlD4HsZJ8kW3vNWrUkKHDhvk+jQK70Ei6+MUXX+gaH75HPqFPPvlEs123atXK983AwABA6OvfdttfjMKNCtrkjTff9L2LGkwI0M0336zJKw8cjNnABgqEG2HlqBSUScfPO+p1jBFJMatXr66KI1z46quvVEYaNWqk5QG8j+eff17rR3JPjmGhfF58p0/fPtKhY0f55ptv9PuBAKKFsuE+3M+C42hQ2FwfOHCg72oU8HpIPsqC/UBB+oqaNWvqkTgWyCTH2Iz/dbzWncOWLTBCfNanT58rHp9zNaDc9957T2bNmiVPPvmkKiqAAeVcQBwSxohNbxEsvv/+e/n444/VqD3++OOyevVq3yei5OCxxx5TXRBMXTDMyAGJVK3XiaEjLxmGgbQdHY0cWGBQaGt7bBBAMZPFnja9++67Y/RerxadOnVSg8kOVerJhg0S0fIcpUqV0uz+XH/22Wf1zEZk2r89rhb0EWlDvv32W9+VPwBh5t6WNCOzGErGCslro5ONfwJOKb9HvtHXnPsIkBuOPGLHLYaZdCoA2bFlNW3aNGCPHz2fK1cuTZjrT6SRe2SF9Cn0b8FChZRQQ6TIFYdesvohELCLmTKpl78jxG5iCGH0/hk8eLCR5zYqMw1ua6DnfcY2BgwYoOVCtKmn1Rn0M4evQ0LZ9UzfQvCxjRAnDmqPa6AbW3/0kepVq08Z7+g1bAUEx+YctPjxxx+1j9FP6EXqiz2DnKPv0VfsRI8z223YcKzBeE2e6VRvyJAh+t4YMq9ChQqe8YL0vYVpBK9du3b6t1GynulszwiHN3ToUC979uzerl279LNA0bt3b69u3bqeUYS+KzcuzKD2ihYt6pUtW9Z3xfOMQvRefe1VzwikZ7xYzwil75PQQLsZJal/G+XrFSxY0DOEQN+bgavPYAisvg8VRhlqeUZheUY5eEZheobUemageF9++aVnjLZnPEYvc+bMnvGSPUMgPEMEvU2bN2udjTHyVq5c6bvb1cOQIM8oF88odX1Pecgd741B8EqXLu3t3r1bP+MZHn7kYZVF46HptasF9eJ16623eoY8+K56njHmnjEs+rchd54htvo3Y4b6UedQYBSKZzwvb9iwYfr+rbfe8gxp8YyR02fp1q2bXjcG0fv888/172BAezHGjKLT98ZD9Nq2bat/047IiiFg+j5YoD8qVarkGZKk70eNGuUZwqD9YrxUL1u2bJ4hC/oZbV2nTp3LzwD4Ljpn48aNKss///yz75PAMGXqFK9QoUKX+wpdZUi9lo0MGkPmGfKpMmIMg+qdkiVLBty+dhw/+uij3ksvvaR/W5w8edIzTotnHEpvwoQJKi/IFXVDDxQoUMAzDpTv2/8MxnPXrl094+jo81arVs0z5MQzZMQzHv3lZ2e8N2nSxNu5c6dnyJL+zpBPzxg8fY5AwX0Yu4xnC+plnGdvzZo1l3U2/Wnbg3p26NBB/w4UPCtycfDgQd+VKHzwwQc6Lixow2LFinlff/2VfpfyYxuGmGqbG0KgzwmsPAP0YOfOnfVv+hYd2bdvX8+QSr0Wl0BXG3KjsmIIkI5JQ3A8Q4g840CoDkVWouPChQv6L/qjQYMGqtsZ38a50uvAECvPODq+d9cWsRoxWvTbImXbNWvV1Pc7tm9X9s41Ijp45Yb8qHdExAPg3REBIccRHgT/kmsIcGgr3im/4W9ABup+/frJd999p9EMvBsLWDbRJ5jnlUBUhGjVl507y/oNG5S1w8ivJzAPO/rn0eohJ0ueXK/hWbRv317GjR2nmb3J1I0XGw7QbnhtgDPmSGpJOB2Qx4k+tJ+HCpJwPv3003o/0iYYY6BlUC5eA3mh2F140003qef4yy+/6PPky5tXs44jG/OMlxEoyHVFtNLCGFspXLiwXjPkXduSY2IAXnupkqXUgwt06ol68SIflv+uRyIayDkwxEKnfInoEJEgglOiRHH1ZI1i1u8ECjwtY3y0noBxQL8S8dq8ZYsYwqLXqTNjLlgwPsn9ZRS1eomEyIkM005EVkiKSptGn9oKBLSd1QGA+5OBHtkgasFRQzaEb9vaXz6JNnHsD3LL7lP6N1Dw/GPHjFUv10a+KBuvmGOEiGIVL15ccuTIoZEeznxEhmhnznIMBHYc03f+ud6AIXU61tGX6ADagCkYzn+kL3kmrgcC9AqRIvQkY5HfUy7lMG1DVn+ii3jytKEhEzpGiVoYZy0sa0SIhBBFoP2wC0QemS6kHxn36H2uW7kNF4wN9P0VBSKO2JQDBw5qpJLcduGo35WAjRv8/WCNwvEs9nmQKcCULMsJrAzR3kSKmGIiWhdK9DMcIP8fYwp5R28hj+gToo3xE8TXyBt1IxrkD3Qhu8Jfe+01HbvodPS5jSwRFeV+7BiPC8QqMfp1/K9auQXzF2hIlgHWslUrnY8nZIpCXbz4N20kGpfvEjplYDKtQY4hslvTWCj15s2b62DFSBOyZj78o48+0rPUmIpAiO1ZZSgqQnscOHslMB3EVALhyBw5c0oLUybPGawxig3wLIQaa9WMCtVf8BkA2ozppKefeVqJHfP14QJkBcHGuBLqRnD9CUF0ZRIODBs2TOejGegoQxQwBg6wpoPz8BgkKFBr7GkDFHhM61r+CdHrgLxYg4fS597IKcQZ5YS8EiYOF/lEdmlnQF3pZ4gQIWWO0pkzZ64eVvzDDz/odwIFJBOFDlkB585HKXfqGd/UwSYipWycCeoaDGgPDCVrfTCcvDhLEOcCw0Zf4cjcfvvtsmLFCt+vQgPtY50dyCSEhGz5FjHJJ/qD9kR+opONqwHkGNLBEUeMB2SBtrOEjSnDQoUK6TVAezCdxjSVbetAEb0erInB6GBMGCO0Pf1sxwN9i1xBzK4WyLqVQ+oFca5/W31tI3RK5iyZlYThzGKIaXd+A5YsWSLp0qfT/g4VEE/kFccIQoLuwfFD79DGjE10O8sJAiV+VwvKYrxzAsJLL72k04VM4TO9ExtAhlhGcFv925QQUv7ZaCQMp5hlBdhHwPob1lyx/IRntdObcQHWNyFzTMHSJ+gw/uVUC/ICPtrkUf0Mwoteiw70O/WG+LFswB/IOdOtvOICsUqMmNO9u1EjFWw8EpRljerVdf0KDBMBTJQosSpTBiJKHCPEvCpkAEOJcDJIUL7cg9fjxmvjXz7HW2R+nDLwBK2niOeKQKE0AQyU8jH4fEYnMm/OfVB4lMMAwHvn2vWCCYYEIlQQAwwLxIE5XVDl5iryXrP3NFqGRxedlQcLIjV4JihGiCPtEoyXHQhQvhgS6uIfwWANANEVZAIjcMkYC0tcMRxKVvRd6PAnfxAjlBIGkYOGaXO8O4yT//dCga0HdaA8FIpdfIpjQJsg98EApwOvknHEOr12bdtJtmzZtA0p19YBY4iBDYXwYdRYS1DROCGMKRwS6kFZGDISrmJMWVgZTtA/GAaiHP9EPog4Is+MEZytQACxw2GDQLDoH5mkjhaQESLY6DQLxuuLL76oxpwoYKjOBE4j+g6DwzNgkCCe6EsLot1EjapVq+a7EhiIkKJfnnj8CX2PnsycKYoYEanyX+iLM4Ixe/qpp5WohgraB0PJehqMIQbVbpxBp0P2kGXGiF0rF1uAZDN+iNLgjNHfsQHqgs2DfN16661KNN995x1tawv+xj75g/ZA1p548okrLs6/FmCNG30CMWZMEWhgHR+6jOgP/VitWnVdf0d0KDqQG6LKLMK20XmAfkLGcULC5YgGilgjRrBAiMkzRmkh1AgZXgeeFQKPoYOsEClgQSQg6pMvX16pXLmSGmLr7eLtMWBtOBFyhWKnYwiTo4QJgdqzzgAkCNJkPUmMLRElGDrlQDIQTLxIAPng+SBt1xP2GsGHhaMUYd54vOyi8PfwqTdGL1whX8KaY8aM0QW13Jcpn7+LvIUDKEUW4jGFZo0O9caYQnytV1rQ9KmdLrWRrFAWKFtQZxuWRm4ZnNwXQ4qMYvDZEcPmAAxGqKA8DCqAWEAiiDgw7WTJPQYOh+GiIU7BANlmgSsLgJMlS6YheDxPiBgvwJQ2bU/5wQCZa9eunU7HMM2JYWaMEVlAQUJYcFqoC1Mi4QLP3bt3b13YeTXTK7QjcgSJso7F1YLvI3OMQaZ00DuUbYHSR3Y47NqCulNnFgvbz0MBugqSBUl74403dIzwLBhTwDICdALTFsEAXYvOhMTSVsgHi7oZ+0wz03acWQn4jHHJ9KzV3cEAObeyjp6mfnanKe1lDaMlJnwXfRTsgmPqFd3Qcs2Cv5mmXbBgvpaPbDM27DFR4QazH2weoR8ZM4wdHFF7hBVkceasmZftGjqJ9kHvMZb27d0X6w7r3wFCw8wNziwRdcYhMzhEgXBasMvoZ+oB4UWfQ3gg+eg86gEYK9a2A36LExoswQ8HYo0Y4V0wxUWD+IPGQvkziPFkUZwMbjwDBmatWrUNQUmlkRsGpBVQGooGplEhMSh87sMLgwph4F40NgOXUKN/iJcQLcoDxcKOJDqJ+zHQ8L7oUAYBgyNcIf9w4OGHH9ZnhhThgeL5dujQQQ22XfeD18w0WzgHMO2KMqatiWBAGi0YoNawhgrkAKXM/YgMYkSJINLnKHnIQ86cOXQNCzKF94R3vn7deiUoKDqiFYGC8qiHBTtQmLJD3oiQIUsYUbxwprMwhBgCplpZzxEoorcZ/Ur0EtCPRDYxPvTvjz8O0evUDxlO4DMewcCuE2Ecsj6GMYkCwzkAyLpVxMGCPmRcpjDkCk+fetJWyBDT5wBZClbRcT+rYAGkiOkkysyQIb2SVf9oRvS2pq6WhB4w/0IUAwHGCnKMd0y5yJv/blmmNOg3ZJdnxHhRX8B6FfoQ3RIoqIeVUWQOMoIRZU0hfchzMVZG//yz9DRtgA6YO3eORukCmV6G8LcxThB1WLdurZIj9Kl1PABReouOn38uCxcukly5cqph7Nq1q++TqwckmvFGpIByMKYcIE4UivutWbNaCSA2gXHx1ltvyVdff6V1s8slrhboeuQDB9vuuAPUEfnk+kpfFIqZA57l008/USLIFHBsGWiiqDjvtDvjHxkh0mijn+iHHNlz6PMA5J0xfN99jdWBZ82jf5TyWoN1ezha6BYcPRxJrhEIIcqIY82OUT6nHelLIkoQXZwMdC62G6eNaVoL+gOyRKQ+zmAGclhhFJLuyqhSpYpnlKM3b9483ydRYPcEu2LuvPNOXW1frlw5zyhu74svvtDV9qxuN0ZRdwcYgfSMIvKMx6Wr9g2r9Aw50HvyuREmz3iquqOtTJkyutvEePu6gt8oEu/DFh/qqvmYQBnPPvus99RTT3nGAGr5hjDpynp2TFyPoH7GQ9O/DWHROtOWrP435E6vhwNGYXhdv+rqmUGou9L8YUiYtjufGeXvuxo8Vq9e7dWqVcszA0d34NgdTMYL8QzR88zA0/LoT2OA9DNjILyXX35Zd5PwPIGCHUpGAeoOSdrRgp0eyANywS4dfxjSpmUGs8Nq4cKFnlFgnjHIKrfAGC7PGFe9J/KHjINNmzapTLPz56WXX/K2bwuujY2x0ToYRaQ7efzHgSFDOp6o54emHMZsKBgzZozuoGInHeUxxoExfiqv7PwxxjbG3Sn/BHZHvfrqq7pTCD0B2GVmlLDuakI2/HdNGpKt/cqOPHa6AKOAdccYfUsfG2Ov14MBdUVObZuhR1555RXPOHf6nnZv2bKljk10WfS2v1ogJ4b0qM5bvHix7+ofYPwZw6nywvgxBlXbg7ZgB+LV1hF9yQ5UdC9jjd8b0q673YyBUlmk7+gD2pjxxu5NXnyff9lVGijQV+gPYyQv7/zi3x07dmidDJHVazwfO/0M2dRdS8HIKm3BmKc86mWBbKHrefn3kXGO9BkYk3znWoC60yboBQvj/P5pFx1137xls+6ApC623a4H0H48q+0f6kEbsruOnbCAvtywYYOOEfoZeeJ99Dam3shFXCJOjgTBCyIsGuj8IVEEvFD7Ox6dF/fi30DvB5g2gbnj7eEdmY69zNivd9COPC9tEk7gwXFv2uRagDrQt7S7Da3/E/CeqPfVfv9qQd25Jx5LbAOZpTx/mQZcp36BtEd0cA/C7dzDf7rAQmXdfCdxmGQHeaEujCX/unCdsuyC3bgAbUHUgHaOqS1CBX0VvX7IM+0Q7rF5rXGlfnVw+DfDnZXm4ODg4ODg4OBDrK0xcnBwcHBwcHC40eCIkYODg4ODg4ODD44YOTg4ODg4ODj4EOfEiCVO18MyJxaIstDQ4Q9cL30TE2Lrua51nf+uPK6HQyZZ3B4TwllP7nWlcgCf/d3nDtcvXL9dG5B1nBw+FmxbJ8WMfxJPQLJRToUghc31Ap6F58KOAtLJkG7HpgWJCeTiIg0NOY0sSP3BUUa8SMtCKp24QKwuviaxGUkc2eFDAjkajZwNNm0+IHfNwIEDtUHIaWAzy7Ijih0knBlDFs1AQVnkqSAbJ/kSrpQEkEFPjotly5bKqVOnNWdFqHldYgvktSHvBxmiAXUkkSaCxe4jkpMFmuPjSti0aZMmzaTfyEFB/htEhb4hTws5N8jCGw7w/OSIIYkeZ2DZpJyUQ94W+o7r9ugCcsOQuwlwunn0k5v/CcgagxLZo83sziEGIzmFyKtBviR/OWVXE0kvydtBIsZAQO4u6oI8k9/HAsVHDhwyxCJzdgca7cy44XPan5wgwYBdadNnzJAN69frc5N/h7HIjimOqiCPDHlwgk2YZ0HuJ/qDLLzk0iEpoN3VR1lkKiZLM2Of5wgE/J7npM9JMmrzAdE25Jci1wnJJe34Jl8Rbc14sElJ2VVFBmESM5LkskqVKno9GHBsC7mfKJPxR/9hBOg7dBYZ48nlQvuSA4ysv2Q1DnSXI7tlqQf1R0ajg3FBDhvy4GA4yW2EvmWsUu9AykNOyPtGPjFy45D5nzHAbjtyz5CQlwzQPAe70zCANk8TMo1+LleunL53CA6MEdq9b79+cqfRPehD7CLjhgzRI0eNlKxZssqXX34pq4wM3nvPPZoUkvx7cZnLCDC+OMYFEgORQzbIQYUNgTCRgZ+8Rf6A2LVt21YdP8YPufrQTyR+JrcUCWLRK+QZZCxdc0CMYgvkbzHGTHOmcGpwpcqVvTffeusv+RdMw10+zZ18QsYIe926d9NcJOTpMQNUPwsE/IZ8I0ZJXT6VOyYYhaInARvS4RnSoflm/HNdXC8wCtKrX7++5iqxIE+LGUx6OjF5eQ4d/uNk4lBA/on77r/PM8rSe/PNN71G9zTSXBP0G6cdZ82aNajcJTGB/BfvvPOOlkPbUx9jcDS/EXVCdgoXLqwnmpMHg74h31Snzp28Vh99pLmMzMD03e3qQG4N2tL/dH3uW7deXa9X717e66+/rvl4/OXUDFwvTZo0AZ+ub4i3N3/+fC9fvnx6ErzF0qVLvRo1a3hG4XkN72p4+bR7YAy+5jwyilFPUw8WnOD/3HPPeWPHjtU8PpxSDyjrkYcf1jxDtANtHQo4FZ0xTN4bQyi90aNH63Xar3Xr1jq2OUGcfg0U5HahDuQmsuPYeKHad/Q97Upb2XYi1xU5fchtZtGjRw/NKcRzGEWreWCCAfltqB/6AowbN07ztRmypTKJruPkf66Tm4oX18g/FijINUUuJHIkRQfj0hBE7VfA6eaUTy4wrgd6InmfPn1UDozz4RlC5H399dd6nXYj1xV6hvxx6Gb60BAk/T5lMjbJe+QQPNBfyCd998uYMd4FM27Ir4ZuJKdcnjx5dPwAclahiwD9To6puMa4cWO9ggULqL6mLsa59IYNH6afofPIQWbztFm0adPG27Vrl44p8nU1adJEc0198803an/Qy+Ry69Spk+8X1xaxSoxI0mS8a2/69On6vk/fvmrYSU7lDww7idcABpfEjoCBT+IxDFkwwACgDK+UWA6jhaKzjd+vXz9VesEQsdgGAgNJJGmmBQacJIDhJnLGC1FCAhBe+swaO0AboUzDAZJ/YVABpIP6cY3EkjaZovFaVTlAJngOyj906JAaSp4tUEMASLxH4j9LjKgPhgggbzfddNPl5JYop7sb3a1yidELBMgYwHj7EyPjYV1uYwg59aAfKbNAgQJmzASeSNIfKBnIpCVyjAXkh8RqRYsW9UYYRwWQmBEHIljQflWrVvV6++TBeIbe559/rn936NhBP9u9e7e+Dxa0OX1uiRFGwib7hCSgI9asWaPvAUSlffv2vnfe5QSJjGv6OHryzqtFr169lMxCCAB6DZIGaFeSIfIv8mI8ZTUSPEv37t31O1cLKzOQcxIr+gMDQ9JbyB99i46FjEEMcV4oDyfjaoHxZUwxzgHJIbkHBgv5xDgB5AdZwYGk3nbcNG3aVA2hQ/Ag0TFEHjs43Djq/kDn0Q+W+EOgcCBx6ElGDIGOS5CIFHlkvBHI4DlJ7mhtKAlBy5cvr+PCH/426+OPP1ZZh3RbW41TxXiKnmD4WiFW1xiN/nm0pEyVUooVL6ZTMAMHDNDpF8LfhMvebdZM3njjdZ0+Y+oCcNSHMRIa3iX8zTSDTYnOlBdnVnEYJCFewOGphO04Dfnll1++PB9rBq6G4Gz4NyYwdUP4mVAlIUpS4RPeIwx/PWHW7Fn6nEwJxvNNtxiFpmH8EcNH6HQI4e1wgTbkZeRDQ5mEypn/tgh0WuDvwFEjJUqU0CMMqA9TFFwjNG+nEDg9HJlgmpCQP9MHTJ/xnqmmNX4Hel4tmPrwT1jHlCtp+QHTt9yXc/1oAw4Bff6551UWmXoNBLYM2sy/POTWTvUwvUQ4makT45XrMReE1jlHiXByMDCKSdcrEOYGHA1CqJspKKaA8vimKzmCgCln+71AwTQk0zdtWrfWA28pg0OIWR/Qs0dPPf9p0KBBOpYDbTsL2s0/0SXT3XZKEtlAp/gfWRNdPjk+g/oxtcVUl/9RQVcLZJMpvWf/96w+C3VBPu05bZzFyBQw0720Kc/FmOG7nIgeCK4kM8ZY6PllnJ1ldSTHAKE7kVem8tCVyO/Vgikz7sc4B0yZIStMZXCdQ5RZ4mCcVJ0OYQxQb/odnc40OOcbOgQHjh9i7LM8gnH5R29HHcJuHHuduj1yNMqucegqut+Qcz02y+qsuABjCvngaBp7kDDyikxZG8pSD/SDteEWjA/APWiDxx9/XOXd2mqmvdHxgchyOBGrxGjK5CmSIH4CNd4QDowdBATj9sGHH0rz5s11Pp41GygU1hGgpFEmDEqMJXOPzGNzVtUAQ6wYnJnNbzZu3KhrYDAekCUannUZtoO4F/Obf7emAXKFMuEAO856QdlBlq4nsD5lwq8T9NwulOAl32JcDAFrAqZNn6brKzhDh4EVDmCwaRvOQWJNE33iv0YLshBujBs/Tg8jZK0E6yv8jSEkAkOA0j5pZMIaC/udYJ4n+m8wOrY8YMvgmapXr355XYr/d0IBsmYNOPdExiFDkKPy5SsYecyhss35UcGANWGMK84R+3357zJn7hw1ZigiyrYkhXJxIngFC84hxLDiWNBHyMrChQt1DRckhPfG09XzxsIB2sv2D7LBmiZ/xRuTPLBWo0uXLqoz/m5BaEygHiwShfhBQKwM8Az2OThvDIJmDQLE0J44bs/ECxTR68HYYNzXr19f5dVmpqcPAUSMsdOwYUN9fzWgLnaNHc4QxhgjBVi/R1n0K3rVrq2zdUaPs77FrvtyCAzIVdt27bT/WL+IzODM2/VbrE/r0aOHEtAB/QfoNUgEhOgJ43y8/c47Kv9xBc6041lxyrFF2F/shQXOBLbj1Vdf9V35KzgQHf3qv/YS4JRCitBjcYFYI0Z0Okz3pZdf1oWlkCC8YYw3h6B+YJhvWsMa16xZezlaREMySDks9eTJk3p4IN4Li7ggRUSFUAxPNW2qg5bDTSEEDFoW07J4zQLvjkFPNAKglBnwLO6yChovjxOc8YLsAm2e4XoCxJDn5RBVDCULIRFIG0koUriIfoYStQdlhgo8XJQhh/sSpWNRMoM0NnFf4/t0QR4eKFFDCwYchIEoIUApW4PBv8hTfJ+iDgX+9wWQFmtMiXbgNTNYIfYosFBhjQvQOhhZxRNkkTcHlXJqOeOAtuDojkBBNI0+JLo2fdp0dVJwEnBEIEW2rpSHMbfGMRjQb4zVL01btW/fXmWW+jEuH3vsMSX1OB8Y0nACI44D9Nxzz/muXBl41ugQyAuLVgMBEUMIDvoGD5lot//BqURzcWC4twVRI76Lx0+56LNQQPSPQzmJruOhQ8TQB3jWgH5ED/hvXggU1khZJ4B+xLNnITCbEtDb/qBsokWWKDoEBiJ+xY39Qc9AjIi2ou/QtxYshsepsPLDwbYQ336mr+sYe9evXz+9HhfAGaLv0Y/YemSU5wcELiDyBDKwq575LzqIQqLzsOvAOmtcY0z5j6drjViTaEgH4fzGxsgyUO3OMgYTDBlPA+8NZVmhQgX9DOYJUWGXDDtx2O4HaGQUuT3VnA7hPpAvSBXRDV7+u3cw6DSs3VnEPSFTfN+SJTxHvE2Ap4VgEvq/nsCzEmYlrIiSwoARXbNeIsBbpp7WgwwVhDPZZYBQHzXtwsnH/u2C0YsNZYgcYMhtPVAYRGw+/PBDHVwokpo1aujuPBQFL2Qs0F1iFv51QB7xeAD3Z3ASAqbupUuX1mgdbU4/ECoOBv7lUVeMHID44XXjCUL87XVIDPU2wq/vAwV9hjOCN0ZECJICQaAeyDvAG2U8BTt9jEGGGBEZetkoOJQ44xZPj745dz5qio4yAt096I/o8oYi/umnnzSKg+z7R3qRz+jTaQAdVMsYExtVvlpQNxwwfs8YpP0YgxY4K+in6LqDPsWI8Sz+4zUQ2HozJiBETAXiLDJGqbedQoS0QT6RWfrWGpmrBSSLukHGkX2i9Rg5xgXTNp07d1YdS58C6kz0kedxCA6M79atW+v0KGQd+XnjzTeVmEJEcfbZ6Qn5txEVximOGTLOlKa/HF5rEExAP1tCjr1t0aKFBjJY3pIwYQLjXC9WR++3Rb+pfoXYYbfZvcbYZUxBvJs1a6ZkCPB76gxJjysk+Ij9fmEG3hMdzgB7+OGH/7SFHKNDQzCHjSfFQGPwM/DbtWunkQk6nOsMRrxcvss6BQSJRiWcSGSBsCPRnm7duimpImIECWM6guk1DI0lPnQApIstwyhtyBYeHVM0GB+8Qsrhd9GVcFyC+jFQUM4QAyJhRDGYqiAiQLsR5mfdhfX0QoVl7H369JGDhiyidGkbwPQkUROUM2UGq/AtGOQYb/qbrdCE81944QUlRUSumLrYuHGD6Z/OamAgvxBqymXqgDYhmhRoxGPw4MFaHlO2GDDkhqlDZIHwNIYHQ4RCol2RE6ZEMJAQpUBAdA+FwDNivFBuKDQ8Kogg287pX+rL2Pnll19UyaA8kGkrw4EAQ01fMS6Q7SaPNlEDR9m0MU4BZI9oJEqJqeRggILGCUJmkFXaFAeGtWJTp02VTRs3aTnU5ZlnntH1hYEC5wlDwXjmOZF96hK1FmulkZ9OupaxeLHi2tZEc+g/29ZEOlC2EOnZpg95DuT3akEfIQO8IJiQScYEdYeAEGnhM3QV79lKj27ib6JayKx1xq4WlNOzZ0/Vd0xnobNYz4ScMO5w/JBF5PM548TgfVNX2pnyIWRXOyY6duyoY5BxMHHiBOnVq7fel7FJf6I3IULAfy0oEWr0u0NwsAQee8ML3c4yBmxVp86dZPiw4bpGBwKCngKQJ/oekoENZCt8uFK0BAqe2ep/ggrUBdnEUUK/HDhwUObOnaf1ZLwSCUPnVahYQccMuhvyBwFnPBJdBTg92DXuFVcIOzFCITOY8CRRBhgc/3lCvEYaFK+GEDsK7IEHHtBOxviiOPFqMRx4MDQexoGoCI14/sJ5VRR43Cg6FDJKGGWM0uD+CBNlIkSEtGNSEBg/Og+ixOI3vC8MRFwJ2dUAwWPQoCDx6CAPEDze33ffffp5OICQM42G0L/zzjtRUQsfKI8+o59o21DbC08fgsNcNP2E0aIPIbcMKOQAmWLgICcYRrxUFAMeK95IoJEIyAekAdm00UxkjzVwyA5yR8Qs+toJIkXItH97XA1wBjCSEAfaDFmj/YiiEB3C8BIVpP/w0DGKhKX5Dp4YBi9QsAYFRYuMQ7gaNWrk+yRqWglytMm0OZ/ZRe7BgLHMdApKjzGNPLD4mr6sXq269hMGlGhtMIoOwozM0XboAPqLCBt9Qb8TTOM67QSJhtAjL1Y+aWvaAPmCZDBOQgnRU19kELKAfNLO1I8NEMgQZfO8EFJ0E4bOkolAgMdsySb1iC7jlIO80j67TNvzHeQHWcEIodeuBuhQ9DXtijGKFy++1gVvnXZCF0AEL3mXdMrSRkvRmbQBv3MIHYwX7Br9jd696867pEmTJjpFbWdKALJOVBD7R36s68VeYWux09QDUo1tZ+kKEXeIHeOQujH+MqTPoDLKsha+w3epuwX1tRGyuMJ/9nR9Qn0oBRa3oeAcHP5tYGg72XZwcHAIDNfPnNE1BF4QHi6hO2c4HP6tcLLt4ODgEDj+sxEjBwcHBwcHB4fo+E9GjBwcHBwcHBwcYkKs7EoLBCxeZKE2/7J4MJzhfxa8/tvWWbDgOFyLrAMFbcmCUBYHsgjVgnZmMTjP5X89EHAPXtF/T44fyo1+HZkBwZYHKC8m2YhJZlgkzrVQ2j6m8lg8y7WY6kFfs/kgmMXX1CGmesR0LRzgWZEBnjWm+9Nf9KXdxRIsWJhOm0XfUMH1v7t/OOuNrroauQvXWKVdeX7uxb+UT13C3Y82n5Y/uMaYp739y7vSeL0a8DsWdNsNFnbHqwWfkaWcTQncn4Xe7IJjQwg7HKN//59AP7DDjvLYWGHHEzLJBgV2LLKo2WLzls2acoKt8nYx9LUAOy/pWzZ3sEGETSlswuDZWQ/LAmYrT3ZXNp+x8D6uQRuSW4tND/YZ6Wc2THHANCkyeP7oYBcpv1u4aKHWw7Y1fY4M2NQU1xqxSozY7UDiKv5llwXbbNld4d9Adisz201ZmU5nsx1729ZtmrEXBJOrAaEi2zbbZclhFH2H0Y0IFAMJvUj6BTAQI38aqduZ+YydAewsCRYowVEjR2kWanbykMXY7kKh/9jOzo4gdg3YcthaSfkMVAZFMEYc+WC78E8/jZBs2bJf3krNgEI8SajHva0c0LfDRwzXZJzsKAt0Vxq7iMjQyvEYtJkdyOxoYqspJMgqG4wSskkOF+rOMwQqS+zMsgnc7A6TSxcvyYSJE/TYHJQHO+2ssUc5UrepU6bKyVMnJV/efHr9asBvqRfPzP1oHwDBYlssaQrY/m8NGqkz6D9SYJAbDAUcqKJFedNu/J42xKDZutCWtDX3J+cOhi1YsNOL7b7IOsaTtqQea9et1ZQHKFju75+niOSYpJegj4NNSeAPUhOQToHdg8gG9cag0baUz844dgqxZZ50I4cPH5KbbioWlHFlTCE3K1aukAwZM+iYQ1dyXz5DZqzRwDCRDwe5gSSyW+xqiRNGmDxG3Jt2pf+pB7sMkQ/qxk47xiD6gPekK0BuqGug8oKMkYqAHa/s2GN3pj/Y/cbYZxcqdSF9B1mQGeekU0H/xWRkrwTGHWWRmoJ7Wp3GOCHlAeTC7tpExrAbpF4YOmyoLFi4QOrX+2PHVGyB5LGkdbi1bl3Ny0YKBdLXIOuDBg3SBKM8I/0CaSNfEPqZHYTB6Nxwgv6kzxgb7Dpj7EOm27Rpo+kj2FlHnaLLIzty2QnODlnSS/jrcewB/UBW/UCJcDgQON0PAHg3ZAxGKVK5AQMHarI+DLoFDJ6BSeMAGqtVq1bCoTEwRrb70fCBgPuTowgvgKycb731lu+TGxcQSDKEDjeGwQKD2m9AP1VaKMZQPUju07tvb/3X5i0BpAVA8PGgbN4fgPJ607Qtypkt3zbZXCBgCzVkD89s9+49ul0fD49t81yHuKAcuG632ZNFPV3adKokODIjUPng/iQmI/+TP7g3OaIWGAJvMXToUJUhtpfyrOTE8ZfffwJeE8/HIMegWMyeM1uv0W6Qd46PsMDwkYsnV+5cUqN6YFvpqRv9xDjzz6DLc5CiApJwwZAnQBoBFBBb2fkueasYc4GA+6LA6RuUIkkXeQHIGElCMaBsv73aLeQxgTYkr1S+fPnkdmNMyeEFKafcd95+R8kQBg+DgbdpMdj080etW+v4CBXIB4rcyg31gmhCDGk/nol/ceyITpAWoXnz99U4BArGH/Vdt3ad1KxRU3LlzKXkBf0IIUJ2MJ4W9COf0Sa0VSC6AHIHScCBhEBAWgDHyUD27Hld5MNC9qknxIvjIPgs0Lb9P3tnARjV0bXhNyHBCe7u7lajUC91o260tJQKdS91p9ShBQoUd3cnQHB3t+Bx9+w/z2QvDXzQJkHb/7798rF7d/fKzJH3nJk5g53A4Z2O3KCXBC4QXv4gZ9wDZJ4dFHC4BCjZAfXv0F8cNj7JAWVOOCcy7IBnrm6IPbL85utvaMrkKTZIPJ+Ii40Ttfi4T+6PNqYvsAuQbvqkWfNmtgQNtdQgpeyZRhvmxOaeaxAo7N+/z96LI3fYSWQKG3666uj4M4qJQkAJfjMHLfCF2XNmWyKeub8uJM4rMcKB8kfFXWoM3WeEE0YOgwc0DhEugkfxNkBDUB+D+iqwSIyR831ABIrBdYSZ33MejAECxXGiMwQHI4bjxHCdCfyea/DHb0mvOsM0lxIoiEUmpWCmuhUIX6mSpWwUhTPNThR1OmBoAwoFWOJBJVYYPO1NrSD6BAV1MhCQpdffeN3WoaCGDAQ3J8CZOZsAs6cOGRmiRKJjroejgyTg7HDkOG4i9bvvvtvWx8A4E01lB7QjjtrJajhAgflLS/1rzzlkhyiaqJbImEid+8sqMAg4SJxZZuOAAcb4XX7Z5dZAQMCQQZwqDvCdd962OnHqPf4TaDeiSIxsZudIZE9Eij46ukMwQmYDo4zTw9mhq9kB58IAInsEIhg4CCT48KMPrYP/6uuvrfycDeh/7AD3X6N6dSs3PB+O44jpkztNNA1phxw4BBSHumHDepUz/e1kBXMKCOfQYUOtTjgZDrIYtB3yyQbWZDIw9JBd5BbbQ62Z7BBpwPcJDumz9z9470StIOSPaxGJoyvICiDLzjPfdfdd6j+gf7bqJkE06BvOi4OiGjFZJ/QeMoR9JAvHH4QbXYB0YyPIZKBLmQl/VkHfnZpFIxNMn2FvkCt8AXYFhwtJ/PDDblaP2rbLyJhnB+je6WQg8zHa3QaY3vfsyUnwRUbpfKL7d92t7NAP2BYCTewbbc49EQC3a5uxJRN9X9wQJOweGTD652ICEoP/fuqpp62OkL0ikKUMzmWGzA4aNMgGuaeCatnIEsOG+DYnMYLtIEB75eWMjWkv1vOdV2LkbHqI8iD0GPwbjHHGgBKlEPXgFEi/U7QKEIWRiieKRfFIWWOAEBBSxRhwUqvOfloIBwqNYSItCklC2GGbgPNQAPJ0QPlROBQdg/C76UyMK87iUgLp9KTkJEsUnX2zMGgQBASTInK029kAsgm5pJ+Izpwol2tDRkqWLGGjf+c4bZQQn6C9pn0hNiszZVmyA4wAjgYCAjlgOBWHgJFwKkwzxIfDQYm4F+SDDCTOEcOF/GQXGObMxMGBJQ2ZjkMmMJY33HC9Ro4cYdsgJ6ldDHPm6+FMKNAHeB7kFgMIIYV8zpo125L7zFmf7ODUCA3gaDJHYFTzdoZBcIL8hgxddoBzo6AjWSMK0gGye8zdoHIvustnPBcyllOgz9gIMhSQN0gj8gHxKm3IdD5DCLE1GFP6C93Gftx3XwdLEs/WwDIcGBsTawMBZIQ/+s8ZGmWolaynM7TL5+gkhC67BR7RQfQOeX/vvfetrnH/7J+HvDtw9i8kO0U/dHq6kx55+JETxDQroM1wyk4WGN1DBpBxAh6G0mhzng87Sz+QaQGQcAgxcnS2wDFyLYr+QQSx4fzxvARs6Mt33/WwspYnd87mnJwu+5D5GLYI24dvwq7i4CHj53OoCj9G2xPkWRn13o9zTbJnZMgonoiNYDpKNaOzefLk1hsmMGWHiYsF+oy2Qi64X/oLG0KgRHKhjNGFw4cP2aE0htkzg76G+JEFw4cjY/wGgs62Qo5enc5GXwicV2JEJA/jx0DQEB07dtQXn39uDRZM8tNPPzOGs7Jxeg2tgkFiYMI0GGQGJYQMAQgRkQyRWDNjkHDizCtgLgUEizFMHGjmzAWfIUxU1jwd6FCMCoSNIZSev/5qO4c06qUCCANGqfOzna2x8uC4DTBokBQ+I0NAlAV5zCkgGQg548QIMmQHRcVZ07atL7vMGP1SdkiNCIEoALJyj4lsaLMXX3rJRpQ5Bf2AISKSIHPiAMNFVoAMDwaTPrbk5QIB0okDWrt2nfbu3Wed7LkA5i/zc+DYaAOyYpBzrkkmAMJ/vpDZ6CBLEI+cGCKyImS/mIMGcDAQLYg7zrzNVVeZqPg7O0xyNoBw0U6QBuQAEKUip46D4/6RR4IyKjfTf0Thp2YnsgPIKdswMC0Aw38q6aQfcV6ZhwwIXBhqxsE6ez5mFZB/SBakHN1jy5vMgQ/z/cisMJQCqNIOcSDSZj5edtvZ6XOyoQzVYKcBQSl9S8ACIE2ZwZZOVDMni3O2wJbzjPQt2W/sDn4CYgkxJHCFPECc8BPnC1SgZ3gN38FzI8vZnUOVVZAxwa8RRDDEzRQGhmnRHQfID4ET/hHCC+mGnEP4ydLgE8/G7p8NyJZC4pm2AsHjNZlwZB97TaD06quvWZJzalaRIVI+JzOG76VPSVAwLIeu4ZNpB859tkFNTnDeiBHOjWgOhw3RYFgLBec4qTNICZvMrVq1Wnfccaf9DRPQEABnSwEiEwgARgeFwQBgeNhdH4VFmHDURC44avZGckAKffbsWTbq5px0GJERv0OhIRx0JhkrjCYGFgOKI+L7lwoglBAWiAoEEGEhje1EhQw9EY1DmoguzhYoHoaJyNcZNqIfrrryKtMvHa2SMlyBY6IsfYOGDW2/cj9nY7BwAkQOGGmMkgOGCzAgDkHI7OBwhijNqY7qXII5JKmpKZo5c8aJYT8c/tnCz0RXjjNH9ngGRwYxGBgWZBu5Pt9EkGsiVzndkw0jho7TVswpwJjRTwQpZADZDuQ6o5sQvZyCNicogjij05+bAIs5Uzhrpx1pJ9oQkKWCIDC3BPvBEER25085wHlhKyAiXJfsEffggAwVDjzzHnpkQdEjhryQaxxBVsG5GH7FVnFNiMfadevsZzhBbBntzXcyg4CGDDgRe3adCW3IcxK5O+elrcn2E+2vM0SMNnRAEEO7ovs51T903SFl2BWeFztCW9GPZBgPHjpoCQN+hMnhZBicIcTs4nT36VzfAc9Lv6Hn2FP2goP0nw9wbUYB8Df0Oc8Mqcis75BqZBjwPfqV7wAydTxT5uHACwkypgQf9BmJCvqL+8Nnk7FFVrEBZI1PbfuMUYID9jUBL58zDI+tILDh95yTv1P76ELgvHkUxwgSheJsaSzAHCM6mBn1RAIwXhoOrF6z2jjhpnaCJelnCAEgi0SDO2P7AOOEQSdiwfDArJ09n5j/MnLkKGMkXrUKTwdxPhSftCxRLMSIbJKzfxMGj/tCCOmUSwUMGXDfzLki+iWbRkYBx5PZQZOOPRtCR1s6Cgc4F6l8+o4sBm1IH/IHGeOPia/AEeqcDDE5/QOIzDBMzkQ8sldkIZhvwXdIa+NoibBI49JPKFFOU/mnZhFQQHvM62gBJLBJk6bGCLSwzpZ7yCkBzXw9SLwzqRPHg8EneqLdifpBvvz57TyHnOB0BtNJdZ9qZtAd2t0Zfs4O6BciPJwpesVQY+DChSpfoYKVKUeXIDBkQXIKdBrbQVaGuWes0KHdcFpcA6OMQ8WQYqwJmsgusAoJGWbvr5w6OIgqE47JYl9xxRV2KoAzJxKQ3SA4c+wYDo7Agf7mfhnec+xfVoDDYSUjskaAWMLofIBXtwgqkRVnf0l0kGd2iBB9jD5kx1lyr5999pntw7vvudvaW5wdz8XegNxPH0PGIM882y4T8DKHpOvLXW3b5iQgIutFwMO/gMDo66+/tjoGuaOvyPSXLFHSPh+jD/Qzw3yQqOyC69BetKkDZIZAHblxMpDYVIbm8RPYWoLQ8wXOD3FGthgSw/493anTCXt2wBCHQ4cP2SAJYOOxfwTL3Dc+luzWqXbsQgE/hG/Cd6L7EHiyuswnhtwxeRwbh41A/yDSJEiQWZIczkpf7AdyBhnF1pMAgAhzPo6dSqouBM7LJrJEUJwWR0sUk3n5LMoAIUIYcQA4OIwmJIXJxC1aZGyOhwIy3ogDxuBAkhBa0toYDQSZzAIGYNr0jDQeww+cH3LDcWa2r1+33johnDzECgeAEcOQ4HQRNhwwf3QgzJ37zUzCLiZ4flbzMOeGe2NpO4LI/ZLap/1IQfPMDG3lFDhHnBpCi6BCJDkf7UA7015EorQRY8D0CcOczAFg3JshJqLY7AoxRojoG6JKFAoxIGMIYSZ6pR/nzZtrFDAji4Vh4F4hJ2QYOYYDzM51eYY//vjDGnrG7jFQAAPPceZxMTyZN0+eE3MfkBuen/bI7qRohhhJFyOzyChthnMlC4pRY9UbxNeZM8JcJq5DlgIHmF3iR5YJ3cHQMhzCNeg/zsc1ce6QFKfNyKAiZ7RFdkH/4EAhLkSHXBvDeIsxmiwzx/jRVvQnE+y5Tk5Atg45Q7+xDRBWhlZwGsgBxyFK6DnygFHF0fOHnEJqcrpbN32B/qGHEAZsEA4TIk270rfIBNfkPaSBIRG+y5AXfUiwl1UQGJANO3TooEJDwqxu4EjoU2wk80sGDx5kM2IEgzgnbACZFa6J3jhzhv4J3C/P0rtPH0tARo4YqdFjRuv666639pv7QLdxcDxDJRPd32qiemR644aNNnu1/8B+3XzTzd4zZg3YL+QSeYDsZHbu9C9tS9/yHBB2+pY+po3JKmQH+A1kB9uCHjvzV5hUja9CbiHy3At9y6pK+hwnzVDahQDPa0l9kyb2HgE6RQIB0gD4DrrLPdPX3C+ZdI5fbBAEQdjJmjp9il7iT0iO0JcE2OgtmWkCBmwHNjUqKtIEO2+dFNgjl9gNvofuX2icly1BnOiOB8cwZ44UEVLmjPAvjYWTw2AhpGQCcHQ4JRoapeQcOAwMLnMwSpcpbYd1IFM4LBqvUeNGVkkRGjrCicRRMAwHynQ6MO7JNWl4OgD2SlQCyciO47tQQLBoW6IrFJisGc/YtGkTc/+tTFvlnFlj6CBEGRmYRqbfWp8wVpAx/oh6cRBOZoG+w1jxPdo5O1GxA/oLo448lK9Q3ioNS/FRGPoR542SIAeQbBwfxJi+4jiykVUn4AASjsPGEDEc6DgtHBAywURC7gPCROTDXBHUBAOEkXIMV1bBeTESEHLnvAAyj4HDMENKeFaux5AP/VCwUEFdcfkV2TbOED6HxJFlhIhBBtEhiCjGluFR2pTnwmlwT9lx3pkBYSfIof+5f/oEmYiIjLC1mGhPovwz6WFW4fQP1yFTgc7SZsgDz+ZvrtPK9Oep/UP/8czO6q6zAdkU7gNbgzwgg9gp7oV+5PkhgRAibBlzhDIPsWUVXIdz+BqdrlWzlnU0DKsguxBsrgNhwemQTaHPeW4icyfjmhWQNaF96EOehT/aCn1GBufNn2f7kme99pprrfzQB8492OMm8DwXbXspwHGHlwLZcHHx4O6V5sKFCxcuXLhw4cWFH7xz4cKFCxcuXLi4ROESIxcuXLhw4cKFCy9cYuTChQsXLly4cOHFeSFGTFu6FKYunXoPTLZ1lmWeCUxSZeK3CxcuXLhw4eL/H87L5GuWebI0mFVFLMFjtQZ1JFgmyVLSUyuong+wYoJVW6zYoN4CqzooEMdqC5aDnw6sSGOpL6tqWB7Lqh3AyqHBgwfb1TBUHWW1iQsXLly4cOHiv4fzkjGCQMC3qMjMknqW41OHguJPHLsQYBkvS8GpYkqhKep7QJRYIn0mkC1iSSylBjKDugxOmfLM21W4cOHChQsXLv5bOG9zjCguR/0SqiFTx4PS54C6QYA6K2wZQl2MUaNG2aqk1LKBgFCEjiwNgNBQo4T6PdQLoTgkBa7IRg0aNEg7zHEH1Nrhc6eOEYSGGkXU5qD+DbtjO1tLkNVy9ubhfIDvzZw1yxY6JFvE/ZD5ogYNWS6OOVkk6uhwLe4NQASpPQL5ciqTAjJXFLXiGHVJXLhw4cKFCxeXLs4bMYIQkLWhACAVWqmuS5E3hrEgOwxJUeCRf8kkUf2XjQI//PBDWwGZAmqBCwNtIUOGvygiRkVZdtCnqBmvGe7q9MwzdgiM8ujsuk0RRIbDGEJzSAwFyiiad8P1GdtrMIcI8kMZdogN11octNiSL/Z04juQJMq1Q6Q4N9+j6BfnpDgg9821eB4yUmy+SNG8NWvX2C0L+B3XYV8f7gsCyDYGEDEXLly4cOHCxaWJC7IqjW0VqNhKlWZeM+eIUv1UXYU0kLmhmjL7G1FaH2JDBqZ/v4xNDKkeS/XsHj162H17mED9xhtv2K1DmLtEduaLL76w12AbCwgJGw5CYjgPxIhy/lRIJotFtophMSpIcy1IENdydlHmd2Sw2K2a+2HvFirbQvY4F4SKjBPXYviNc7FFBUOIVOWmNDrVcRnK6z9ggL0fqhDzHcrNu3DhwoULFy4uTZxXYuQMYbEDM+SBDAv7zxwwpAHCwlAXW4awt5KzrQMTm9k6gf3KID0QD8DnHGd7A4bn+JetEiArnIstLSBekCYIFJkn3jtwvkfWh1L3wLmWMzncuRbfcYbCnK1BMm+Q6lyL52MO0+uvv24382PTQzZBhDRBoLjmAfPM3BNka/jw4SeG8ly4cOHChQsXlx7OGzGCCEAwnD1nICWA47l8fe1x9jbKY4gJgKAAsj4sq2dDTzbahPxAQJzfO+fkj2MM1/EakkU2ivcMu0FceM0f13S+52teZ74WJIa9lpxrAb7v7GfFdwDnNiewryFU7C3E97gW/7LDP2SMfdb4LddnjhL3xXAix9gPy3kOFy5cuHDhwsWlh3O+uz6A2LCjNfN6yJYwhMRqNDY3fOutN+1cI4a+Dh86ZCdms3s4xAEiwU7ukBEmX3/33Xd2GIrvQo7YwHTAgAH2/AyvseM+54ZwQFDIyDBBm5VnbGrIRG/mLjFxGkLC/B4I0qOPPmrJjHMtSBjXYviMuVAQKHbjZi7U2LFj7aRuNkxNTkqy86JYzs+Eca7FCjZKAUB+OB/PC9liU0dW4zEExzmYa8QkbIbr2KTRhQsXLly4cHHp4bzUMYLMQDaYFA0YomJICrLCjtAQIHbKZ6iJOUQQJsgIJIXVXmRbOEbdIzI1EBx+T6aGIS9IDbtrM4eHc3BeMjOQEXbpZ/iOidDM84GwkCFidRrnYo4RRIZrQFQgMmSL2FmcITIme0Oe2Amcc1NmgHtzhvq4X0gcZIlrMZ8I8gOJgswx5Ebm6cjRI6pZo6b9Hd/lvrl/iCDnd+HChQsXLlxcenB313fhwoULFy5cuPDCTV24cOHChQsXLlx44RIjFy5cuHDhwoULLy4oMWLOjgsXLly4cOHCxaWKC0aMmPjMthguXLhw4cKFCxeXKi4YMfrss8/s1houXLhw4cKFCxeXKi4IMaL4YZ8+fexS938Cy+dDQkLsUn9qFznHWJpPnSEW0fGez6OiomxpAGoZ8V1ek5lyhuz4l0KM/I7l+YDfc36OUw+J3/Ga3wL+dV6zBJ9zU0bAXbznwoULFy5c/PdxXgo8ZgYbwvIHwWBvMbb/OBPYhgMCxW72gwYPtnuPUQzyjz/+sMUiJ06caLcVqVOnjj3G3mrQlc7PPmtrEbFZ7U8//WQLK7KlyIA/B2ja1Gl2Q1nqEbVt29a+/vnnn20RSI5xXy+88IKtr8R5neagbtK7775rayhRuLFNmzYnKmO7cOHChQsXLv6bOK8ZIypAUwGbbTIgKpCPM4EikGzOGhYeZne+r1+vnnIbckJF6vHjx9v9yNh9n8rX7JNGFmft2rWKi4u1e51BivjXyRZBsIYMHqJXXnnFbjjL5zt37tSCwAV2H7M777zTkh32TaNgJIUYySxBgtjwFQLFViHPPPOMrXTtFmV04cKFCxcu/vs4r94eUsOQ17Bhw2yG5u+2wiB7s2LFCnV6upOtVA2ZcTJDZHSoKL1t2zZLfCAzZJXYuPXKK660BOb2229X3Xp1bUVsKlOzueuTTz5pX0Oi+B1DcB3u62BJEr+H8LDNCCCb1b17d/t7jt166622WjXbkdx00012WxEXLly4cOHCxX8b540YsS8Z22WQ4SE7w7AY+4QxV4f5Qs78IQcMabENSMWKFU/MM2LLDr5/yy232O/07dtXl19+ud22A9LChO7Vq1fbLUFuufUWTZk8xWZ7mCPE1h9khAAE7eqrr7YZLIbcvv/+e0vWOAd7qbFXG1uUsP8Zw23Lly+3vx80aJDNHEHIXLhw4cKFCxf/fZxzYgSRYTNXhr7YF4xMzbRp0ywJWbR4kSUgHTt2tHOFMgNCs23rNnX/trt69+5t5xSxVxrDbyNHjrRkhuGsr776SosWLbLZJPY/mzNnju644w4VL1bczgdijzYmZfNbhsV+++03+/7LL7+0G8TynnuEDJGFYk+zDRs2WPJDpgrS1q9fP5s94h7ZBy0gIMB7ly5cuHDhwoWL/zLO+V5pEKFVq1bZ+TqtWrWyc4vI6kA4yAiR3WGneiZIOxuzArI8S5YuUXhYuEoUL6FWrVtZokLWiEnZTIZm81c2YmWuEHOSIEdM7CYL5MwL4nOGwhhe48/ZEJbjECDug2wQ91KlShWt37Be+/bus9kl7pvvcV0IFoSI++f3bCTrwoULFy5cuPhvw91E1oULFy5cuHDhwgt3qZULFy5cuHDhwoUXLjFy4cKFCxcuXLjw4j9PjJy6Rv8VnLqa71zhdCOqtN2ZRlo5fqbfxMfHn/F3ZwLfP11fnekeaIfsXiMzsvtsp7u37OJ056Ai+5nOzepM2vJswHNQ3uJ0OJv2OxXcK5Xkz3RO5gTyd7bg/Kdrr3PRP1lFVq91rnQ1c5vy+kyyf7ZtcLrfc63T9euZ+iE7ON3v0YczyfzZXu90bfZ34PsJCQlWti8EeL6s3OM/6drFwpnu50xtyPfPtk/PF8575WsHLJWfNWuWatWq5T3yvzh06JBdwbZ9+3bt3r1bBw4csMv3KcCYXezbt08jRoyw231QPfu/ACaPUzDTKUPAhHXKIrAyz6kTld3q3NR4WrBggZ14Xq1aNTvJHWGlyjg1nFhFSPuxgs8B1+3fv7/9HpPYHaxbt05z5s7V8WPH7LmyWhST1YRUMeeanI8J8GDhwoX69NNPFRQUZCfKFy9e3BpOVhv26NFD+/fvU/36Dexk+uxg5syZdtUhiwDYpoZaV5lBnStWMrJQgLIRrJLkj5pclJzIbhuzupFrHj9+3LYLoO3mz5+vCRMm2MUELCyg7QHOiLpeLCbgPpzfZBfI/thx4zTb6B2rQlmpSXFTHA9tysrPevXq2WNnA87Ntj+UuUCHuV/nnBjw+eY5lixdaleRlipVyh7PCTg/RWAXL15s+4ZyILQjNoNVp5s3bz5JVlkAMs48P/JN2/PZ2RZqRWaQP1arYvApPks/scAE/aT4LDXPKP2BTEdHR6lOnbo5buOxY8dae8gCEGSflbW9evWyWx81bNjQPg92kmsht8hO5vbPCjgXK3F79uxpF6cg4/QV8s7zISssZGGRCwtTeNZvvvnGyjRtXalSJe+ZsgYWtrDKGPsMuF9sPHpCfyH73BPn5Tg6iKxu2bLF+o/s9iG+B1tCnTzkxtFfZALZYUUyepAZtOmMGTNs22OT6NfzCWwqK6bpNxYS7dq1y5aUoS2wzSxAwhdS1gbZQhfQu+z29fkCdQHx7w0aNDjhr7Fd+CfkBTub2Yfw2S+//KLff//d2iPsH8+BfuEHeG4KOCMTp/bNhcAFyxjhiBDCvwOOAYWbPHmyFYTnnnvOKkpOEBwcrM8//9wK+H8BKDeFLkeMHOk9Iqu0CBaCBynJCYjiKa9AjSiHvU+fPt2WSMBADh06VC+//LJVXAfUfnr11VcteXWAEuO0MCDUgsqqskLMuDaFNSEITz/9tHUAKAXPhpHAET711FPWaKMws2fPtor01VdfW8KSHUDCKP6JgUTOKB2BA3DAtV566SVLCAGOCNIJKUMecT7ZAW2KEfjoo4+s8XDAa2KS1pe1tg6Ge3KAg6L2FgSR+ls5Bdvj0I5t27WzbUxdLkCkBpHBwdLWZwPORQkNCNFdd91lZQmyB5AtttWBnDVr1uxEMdWcAFmnmj3ygPGkmr5T6BUZpT8JGt5++21LWAByPGbMGCszOOOzJUU46HfeecdeD+A4aVcIGU6b/oRAIJ+sloXMv/76GyfaI7tAp1988UUtWbLEvkfH0BGcDLsDIDeALY64N1bOPv744yeOZxUQFK5B26LrtDPg2XDA9913n3XOkC/sAAETq3XRkQ4dOthnziqQN0gR9gz5eOSRR6zO8frrr7+2xI7CvRA12hfQn8gqfZmTPuSeIXg44szyjj1BRyYb550ZyAvtAPGgCDC6f77Bs3bt2lXHDFkDFDamHyFIyA9tD4GgjSD81AZEDiERFxuQtdffeMP6avoP0HbPP/+8bctrr73WylZm0O7YYoJSnhs7y28JeNlsHn1H3mmHiwJj2M47jCP1GLboMY7Te8TjMQ7DYzraY4yb94jHs2/fPo+JgjybNm2yn7dv395jnJT3U4/HKIj94zMHRug9Jir1GIG3fw6M0/aYDvGYKMF75N+Nb7/91mOUwXP5FVd4j3g8xgh6DGHwGBLpPZIzDBgwwLa1ISn2/fz58237AWNQPIYgnbiGEWbPQw8/5Lnhhhs8RjHtMRMpewxR8RgFtu+zA2TAkFf72kQcnuuvv95jjKY9pzHG9jj/mmjfY4y0vUejiPZ4ly5dTpKprMA4AI9xYvb1xo0b7bMZR2vfG0PpMUrqufzyyz0mevMYwuQxkeqJdnnzzTc9xhGcJLNZxd133+0xDsX7LqPvOnXqZF+baNZjIi2PiWDt9UwUeNZya4iK57rrrrNtCoyj8Vx55ZUeE4nb93PmzLF9HhkZad/nFOgc5zVkzr43hNljHLV9bUiR1cHw8HD7/mwQtCTI9hegbSpUqOAxxtPKhiGu9riJpD116tTxmKDIYwyzxxB0jyGcHuN07ednA57TOCdP+1vae+6//357jPvgHoAhB55HH33Utjv9aMiwtU18l7bPLsJCwzyvvPqK58abbvS8//779pghdx7jPOxrQx48xhHZa6GvPCOfPfTQQx7j1O13sgJkHFlw+ujTTz/13HrrrVbmkVH6ExiHZW2NieCtHgKujb127EBWgK4bx+dJTU217YNN+/DDDz3G2XuuMLaNPgSGMHkMQbOvAd/hfpznzy7Gjh1rZdGxHQ5oW87rgM/5nglkvEfOPw4ePOi55557PI0bN/ZMmDjRtgt9yr/AkAPPB90+sK9pIxMw2tf086+//mpfXyykpaVbvbjuums9DzzwgLWN9O2TTz7peeKJJ048w6kwgfeJzwyB8pigynPkyBGPIVL29+C1116z37sYOO8ZI6IpJ8VNGhaY69oUcffvutssiBOFE50wFkmqk4rZRCRG0e1npKuJVIhAe5hoGpAC7datm9577z2ZTrFpOwdck5Te2USplwqIfGPjYm2UYBrPHiPigqkzTEDNJ4bTzgaZ6zQRJZGiBURoVBuncjkROpHbM52esWl1+hHA8qlJRVROYU/+zSpI2ZMiNkphIyO2YmEogmGC5s2b2+9wbxTkZBiGdCxZKWSF/jfOyH4nq2jdurUdlgDcM8MxzvAumRqiVYZJeFae+ZprrjmRAub71M7KyWbCtGPmNibTwXUA1yerhLwTSfGc9CsZCNolJ0CPqBDPUBYgwic757w/9X5yCjKD9AHZWbJvROSG8NlIl+xRlapVjZ5/ZzN7tGlOcflll5/QZa5J1gQ5QUbYzgfQX8gSeyYyTEIm0AQUuv6G621bnA3QL4YmOz7Z0WYB+WvZsuUJ2WGohqwVWT6iYNqXzBFtTgHa7GLgoIFq3qy52lzV5kQUTgbXOBNrO9EBsolciwwtw8lck9pr6E5WQZsR0SMfgLZjSB6Zx6bSh4Y42GFfQ07s8Bb16QDfQSeycz3sBlMBsM2MEHC/9Cvtdtttt9kMGfLD8z388MPeX51sn3KCM/3esWEOGA4iy8r9kZXjuc8n0FOyQDwzes972gUbzL/YebKP1117nf0+GTYy82TQkUFDPuzxi4Xx48dZPXvwwYesfmMbGXolC4m8MhUCGXJk2IEJGE5MHaCvscsUc0afaXt0jSw0dv9i4LwSI1KgpMgwDAi+M7+gX//+9s9E4LYhwkwjAJwdjclWH6RSaVycMoQHh/Hss89achARGWnToAx53HDDDTZlZ6LJk0gQaUjOTaP/m3Ho8GEtCAxU52c7WyOEMgAMGnMdSEOzXxwGBaN5LkF6G0LKPnS0I8NrDIlQnBM4xoZ7wDlh8DZv2WKHnDIPT2UF69avswrFcAhOLTMwVjghhiYAKWVkgvS+M6yRVeA8HJBaZz89nAJpYJxL+/btbRs75Md5Rsa6mUOCYToXwBw7fQlw9hAj5sYh05dddplNryPjOQH6BvEiBU8fTp4y2T6fY4xOdQhnA8gkzhT5gOjyR1vRT7cZotvS3AfEidR4TkE/OH0BaXzwwQdPmt+GrBJAYCPQDQwqQ8zjJ4xXRHiEHcrMKSCn6BoBGEY7swwB+hH7g27wOYg0NoqhWoaoGHrODtAD9AfCybkzXw+HAeHij3sCTrtgP7G5BBfZgfN79A6Ch06AK664wsri4MGDrXxCmjKDoVm+k505IJn7EdKBPF53XYbTx09Ur1Hdzl+E+DqB9IUCbY1sMU/GISa0BX17vkDbQgqwqehkRsv8BfoE20M7A+wTSQZ8IzbZ0eeLAYbCGNb+4IMPLPFxpk9wnICzRcsWhkhXtDaM4bHTgSCaYTMnwHVkAx0gyCHYuCgwnXHeYJiwTe1+9913HmPEbAqWVDMpQ4Y0gCFFnuSUFJvCJCXP0A1pedL8pE0ZOiO166RV4+LjPQmJiZ5u3bqdGIog3cbQjpOCI0VnhM1jDJN9/28GQwEmSvM89thjHkMOPCZK9nxgnt0ZEgG8btq0qW3bnIChtFtuueXEkJEDY/hsah0wNMEwBWnm++67z1OuXDk7/EOfmqjZ9htgaIP7NVGXfZ8dMExnyK29HwcMLRnS52G4NDPoa9LIPDep6OyC4d033njDDpGEhIRY2bvmmms8xuF6jDLaIQKGKgBy+Oprr+ZoqNDBvffe6/nyyy+97zx26LFXr172NUM/1atX9xjyb48zZAJoB4aMuH5OQJ8xBEsq3pAkT+fOnU+ci3alz892KM04EztkxvDZqNGjPQEBAZ6+ffvatkIunGEs0uoMNZ0tGGIgxc6wZ2YwvNC9e3fvu5PBb2688Ubvu+zj9ddft8ObDO/QjoZIe4wz8H7q8ezcudMOMzn95oBnf+edd+y1GVrLKtAxE9R5Hn74YY8hnFbXnCEmwHAFOsGQrwN01wQknkWLF3mPZB/c67Dhw7zvMmwPOmKclL0HXjtg+LLL8108wQeDvUeyB+w98rhiZcaQMfrN0DjDjoYs2KkXY8aMsZ8BhrbOZigNXaZdTx1KM2T3xFAafoMhLYbtAHaF9h8zdqx9f67BsCJ9iO9iqL1kyZKe5ka+TCDj/UbGELgzrMez00cmoLM+r5DRNWcI+2KAYUh8AnKKHeb+33rrLWuX8d/4bvqVYV+GYk8FUyaQqWnTpnmP/IWPP/74xJDhxcB5yxgRDRDpwP5hw7B/VsUwKZrI48RKBsOS/Q3TZCIvqTgyD2wau2v3LpsRIBIkLQ+Lt183rD49NdVGYqQfyRwx+ZBozUnVE62S0ifa+beDoRwmt7LDP3vPMUmZaAZ27jwvmQCySUTKOQEsnUjXYeuAaJRIiayeM1GSqJ8ULvdCREdGjgwffblly2b7O1Lj3Af3k1XQV4CsA6l2sh2AVSQMG3zyScbCSSYdGuNg/7hfhhBI558ayf4TiAqJYEiV81wMYzFsRXRIdoznIQpmyI0oks9atWxlI1rk12n37MLJJgD6EPkGpMrpV7KhtBuyDWhHhkpyCvqI/qtUuZLNJNB3TvaBf50I72xA+yEr6G2H++6zQ6lz582zky35zBm6Qy7o27MB2wMxaZyJ/7SNM8xIRoahshdeeN5OQo6JjbF95GRQyWBlV0Yyg7Q/QwJkYpB5Uv5O1hQwpIsOogvAkVGOEenTpwz7ZRXGWdhndLIDDNdlHq4im9nQtDcLEABtzJQCJqSTPSbDwPWzCmScVX1c58EHHrBDGAwBMh2BzBvPwFAoQ0zoKllNhtRf7vqySpcqne0FLpybDB5DZS1Me3I9dJupAdhxsgdkGchGOMA2na28nmrjQOZzkn1BjpEzhrToP2xR3hwMnWcFyAXZFoYq8VXYALKrzvAsQLYNWbOvkWl0jT5BJl995RVryy4WkDcWO2C3kE9GhHgOMvD0J3JJVov3jj9w7AFyhN+mvZFzZ+gbsPCKydtkzS8WzstyfYY5cDoMqTBXhRUBpJVNVG6NCil10uwYMVK3NB4EipUdjjGYP2++XZVhmKgd38ch0Zg4EUgWqWTIFL9hyIHr1ESgDNHi2qyUYNgjp2ThUgHthdAhQKTnGW7BiOE8Se1DWmhbnDZtlV1gFEnnMucFEoYzxfmwOgxnQl8yh4hhGRwD9wFhgFiYSNgaTZzeqFGjrRGZPHmSIXBNM+ZDZQEYWYw6ikR/Y/Rx4AwLoPwYroULF9mVLKRn6XNWzWAkuAdIGsY0q+A3DMPgtJctW6rff+9tDSDHMDi0NdfmeZFFVqcwZMD1SF+PHj3GKj/PnFWg6AwFm0DEEiLkHeJAv/Ea/WBFF0MKGORRo0ZZcoYRxPhAiLML+hUdYBiN/n3xpRd1w/UZgQJ6BOFk6Omyyy8/q3Q1zgYHSkocw45+MjTAfJG169ZaOYWYoOddunSx8pwTQJJpC8gOr1nazLUhAcxTxN/NnDnLzhNrauRv9pzZdlUVpJcgjeH2nF6bITvkHtnAqePITbRsnSwEhCFZ+g4yzXuuy9Asht65b2xaVkGAwPX4I8hDr7BlzEXB7vFMDBOio8ghhN6ZAzV48CDbr3fffXeWhlm4X5wz7UnbjhkzWsOHj7S2gOswTM7zQ4ogtgS32AFIKYSQZ0UnGU7NCghkIT7IBfafqRbIKLrOtWjb/IZEIkcMzdetU8eSenSeAJl2duZDZQfoLnLAvCwnKIeIMXSPfqLrkFfsH32H/jDEQx8/9thjVobPNbB1Tl/zL7YWsoj+gI2bNmrO7Dl21RaBDPIOacNm0ga0ET41J3b/XIC2wjYxhYXAhHZkDjC2jbZGfjiOHXrmmWesncBnNW/RXJ99/pn69+tv25c+wAYiQwS62F/kg+khFwvnnBghUDQGrJcOg8TQQMwVglFaA9PIGJioaKvUNAb/4qhwGkRdGDCiIX6LwOD8+Nw3l69VFibA8j0M/PXXX28zTMw/aWuEHsXD0aFAjFFmJ3NxqQOhwfjStigKhgyjQZvgrJ1sQHbA7+kziAAOkgwF7J1+gmzShvQdxtCJrrgOSsF3UFCMPt+DpPEvhiSr0R1EAGcDUabfISg8J0aLPoWEYZghP5Atrk2NE4wE8oUDzg4w6GQfUWYUuGWrlrrr7rtOiuh5JpwgxpA2JnLBmNI+99x7t/19dtoameQZ+B3ncQwwckxbM3+KZc8YCZ6T5cE4Ve4B2c5JpIzDI+rCWOF02l7d1vtJRpaHyI3+q2jup/wpS2mzA+6Z++d6yJFtIxPh0j5Xt7nath96CvHjejkFxBmnTL/hoPkXwsFz0K7oBG2MPSCbgw1Aprg/vsf3zwUItNBBZJNzQ3Z5VnTQcZ7oATYPx8dx/nIKZIF2Q17QE86N7CMXOCWCJZ4VGaVdqlSpqgcfevDEfLx/ArJAO/F7flOhQkWrZ8gh1/XP7a/EpEQVL1Fcjz/2uJVXrsPn6A/6+dDDD6lY0WLeM/49uF/kH+fPebAx3G+1qtXs82CPkKVGjRtZIk/bogu0LddEf7JLjHhG/hwb5xBk2pL+JCAqY9qXQJDgi/fOZxBM2v58gz51CLiT3YyMiLQT3bGpAHnjPTYTXcMHcn/ZsUXnC9wz9goZwn5jE5x+IwuP7tIH+PR6deud8Fv0Bc+ROcDFZnAuh8BeDLibyLpw4cKFCxcuXHhx8ammCxcuXLhw4cLFJQKXGLlw4cKFCxcuXHjhEqN/ORhr5u9cgXMxxn+p41w+88UEz3HqaDbvz2UfXIj+5DmY+3AmnGu5+qfrnQpnXoaL7OGfZDG7/eDCxelwqcnQBdtElolXrDr4u0mBTBRlNjorTZjAzaRVJgNnd3IZk7dY2UNFbSZ/XcxJXOcSrPxhRZ9TeZZVIVSLZnUCEz2dApo5BfswOedjMiAT4zB8bODI0komXjKJzpnYykoCVhPyvnKVynZyYE7AShT2xKGQY+bJlazIYUUcK2KQG2clGCsUx40fpzVr16hihYrZWgoNmBTIM7KikcmYPCtglQwrZJggysRQwPNzPeQSGc5JG9NuLD1m4rcjizgc7oH25jgT2JFzVr9RjHHWzFn2enw/p+3KxGQKndrK6bGx9lkz6xLLrFkV5KyCySmYDM8KO1b8cU0mizrX4Rl4TlalccwpxZBToNvIA9eiDZFRisGx6o/VL8inMwkaMGkbu0PpCb7LKsCzAedhBRiTZJlUzuob9oNitSirs9BDrkG7s9cf91u9evY3r2WFGSvaeCanDACTr9lvkntAdrGRmUsB8Bv2E8NmMgE2q6DPWIlFUV0mxSJz6DR9x3OwOhL5YaK0I4s4MlYZsVyc+/qv2NiLBcinUwA1s45gZ/FjTAqnr/GRrASkX5j8fiH2ccsK2E9y0aJFdhEE9hR7SXFZfBa2LbNOZga2n9+x2o6FVqwERKaQLV6jv478X1AY43JBQJFHCnj9HUwjeYzBsUX2KAhlFNEW4ssuBg8e7Hn66ac9111/nccIzom9uP7NMA7T7iPWuHFj+55CZVdccYXHKIhnzty5nssuu+ys9kyLiorytGnTxhbFDFy40NPanI/Ch5z/3nvvtQXsjGPxvPveu7bQmFFiu48SxdEMiTixr1l2YUiK3YOMwmUUWeQejMOx++WxLxlFPBs0aGD3/aIYoTH+thgaBQU/+/xzW1zMOB/v2bIG42xsATLOk/m3RiGtzH311VfeIxntPnHiRI8hRCcVnMsqaCuKYBqHfdJeaatXr/ZcffXVnilTp9h9gngeYOIUW2COdqdIaU7k3wEFVtE59ktr166dxxhY7ycZe39xXe7rbGCIo+2/t99+2+5Bd8cdd5wo9mmcpy2ESGFEigTSFmeLzz77zO7JRF8B9rlDPp955hmPIWSeZzs/e6KopAkcbB/zG0Nu7bGzAfJHsTpkFKArvKbgKYVsjUOzsmKIhOfZZ5+1e0BR9M5pj+wA+afII4UcHXzzzTeeq666ytpHnovn5RkdUMwTOTUBhfdI1kBBzttuu83TsWNHj3Fgdu8rQAFS9g40xNIWlx04aKA9Tj9S/PH++++3z4osuTg7oKO448lTpniPZOBr0+dly5Y9sZ8jdpKCqtgGEyDbYxcbFBi+1thnCsYiG+x3llknseHY0VNhCJH93k/G9oWFhVk5QpduuOGGE8VUKcp5MXBBiNHs2bNttViMfmZgVPlzADHC8TtOloZ+4YUX7GtAo2f+PuD9qQYXA+38i+HfsDF7huJSBNVOqaALAQI4VqqOHjlyxL5HSUzEal/nBBhiqi6zkS+ApNB+Jjo+0Z4YXoTVsHhb5RcHxB99hoHMCUwkeuL8kAGuC1ni+RYaggYwzFWqVLHVtHEyOCMIEvfBtflddsGzQLwyEyMUE6IF4XLgyGizZs3sRpTZhSOvkJDMxAjHSTVoQBvjBCEPkMCD3kreVJalGndOgFwgL07bUEUWcgQBBlTDxsk3adLEvs8paL8rTJ/hXAEkyalyS6Vv+op+Ohf48acfrUNw2gdgXFd4N9wdOHDgiU1keU7INKToXIB+7N27t+0PjDdAPpFJwDUhR/y7f/9+G1TQNvQ7GxLnBJBaiIkDiLmj7yYit87DAc7opptusve3bt0679F/BvoHwXPOC8G9/fbb7QaxyLwjP+w8ACnj++hgvXp1jd5utp+5ODsQ5CJTdYz9HZ+puj6yTbV4bIJDjJy+hYDXr1//nAQbZwtk44orLj+xiSyy6dwvwRh+5dSq8OhH06ZNrV9zwHfmz59vK2YDZJEkx8XAeZ9jRCqNCr/UaMicTmN4hj3RKOLEkAygsJNxTqJuBKld0nNO1U/S1WyeRyVfUtSAVDIjga+88ootCkg6HzibhFJLhHpIJUvkvHrwpYAFgQtsTRqjPCdS2aTyaVNqtFC8jjo4pLpzCtqMVCbno08oeEZa1DD2E+1JOpe+IU1PepN0OsNffI86FDkBwwCcn3Q8KVSekfoh1Chyqgvznhon/MuwEKlZhtv4LcMWpFyzC9rx1CGqMx0DznBbdpH595nPzVCQMyRKHR6G7xjyYSiDoQ1A7R2G8YxTtu+zA4ZBGCrjX8AQILWhGKpbvWa1tm3fZmtG5fS5HFBE8K4777SVodFNhtU6duxo75vhUWTjj3797BCPMeLeX2UfyH/v33vbIq4DBw2yBfuwFRS4o2YKYAiJ4TLkkyFKhppNo8sQXZuaPxvQN9gyCtXRj/SJIX0nim8yHMBwEsMg/IusYvf4nmPDsguG3zLLDOfh+YBxjCf2GKPNsYlUIaaeTHbaGf2hlpxzXoasGQotYO6fNkceAW3MkDD9SqHJatWqi4KaTrFVFznHl199aWuNVTZt7/QdQ0wMrVEkmeFZB9h9/Br2g8+yO0R7rmGIs5368vjjT1hbw/1T984Znkcn0YlTh3apnO7IFlXQKVqJ7FGAk1pHTHdgGJlK8xcD57VVaSjmoFB8EGVzxkPZbJKd8l959RXVMsTFcWzMw6AhqSr7zTffiB32ccw4TLajoPor1Zb5Po6UDSMpCoUjxzBlJgY0LIL1xJNPnlD6fyOY9zJ71mxb5RZi4jhJiv4hUMzfoIoxDuhsgIJxPpwJTgfykdmZM8eAOQ+0uTNPA8FlbgfVeCG6ZwN+j/NENjD0XNe5NvM4cEDcE0bYOc6/3Pe5LMRlggXvq/ML+tExajwHJJM+hHC+8847trIxc1kwijkxfugRhQhxmKtWr7KGlHkjbMA8aeIkPf3U01YvzoasOGAXdvSbOQWQAkgrpAAdhTxUMASeLR6Y95dTECRB7NoYslzVGFC2zRhkCJIjC5Ak2gtSxlwMCDvzxK4yxOmwcTJUFM7u1hUOmOtINXjsEkb7dP1B1XZskVNtmrkRGH9sF/PWzgWcZ2XeCXrqECP6mOrXzK9ydCKr4PtOAVHmKKFrjz/xuPKbIBbCzjPQl8xzoV2ZC8I8FyreUwySKvzYchc5A/auTu06NrC0NsHbx8gbRJg+4LgTwPAaewvxR/4hqxcLJCYgRgRFADnKLHtO4IxO4rsygwAXIkTwt87oNvoJ4XN+z7xBAv+LNXct+xY3G2CSGCQGxdqyZYs11Bi37t2768MPP1RAoQC7vxJGE2dIxojy/lS9JMKFUEGumPjLaxwHFVpx4CgsjU2VVs6NYSY7BHBuP/zwg/3u/R062GP/VuBM+GOrA7YeoD3ZHgMjT7QGmbjBkMVnDGFh8mdOAfGB5EAm7zYKyfYNtKsDlJD2z1xpGqHm+0QIKPLZgAquGAmcEBNMHbBVCU7A2WUeQ+6QF/7FsTvG5N8Ex8kBDAjPAZmhbyGAPDdZUohFTgB5JbhAl5YtXaaFixbayr+BxlHTZ8gOE9uRGfbiQ/9yCpwmW9Igo999952GDx9unw+C9NBDD+mJJ56wW0ywdU9OAfEh6sSAck4ymhhXB8gOjpvgCUCg0X+q6bPlDFk4JvPnBPQJpIvzoIOQTNrOAQECpCLz1gwYfbbZYPsLtmRBv84VuD7nh/jxmuCTidNsx4ANxVERGGYXdmuT669T61YZ0T73T8TOZHf27ELfyUjhvAhG77vvXtvebGXhIvsgI4fOQB7YYxDSCcnFb/76669WjxgNIZsC+YSIQJDIqhMMo7PI5cUC94n8sa/f0KFDrc2CKDvAb5E5RQdOBfpINXzsQg9jM/BnBBcOCAwh+tldWHOucN6IEdkfjAXKxRANysSDYohpLI4B9lfi4TmOMlOOHSO7Y8cOq5AYOMiAUy4cgx9gDCQZBtLa/IZr8LnjMDHOkCScuDMs8W8Fz8VeOU4pe2ebFNqG1Sls1dHvjz9sFM3Kn5wCQeSPtv/dtCcOlegRYBTJXGEMMfDJyRmbcwKcOSXfMzv67MLpNxxLZmVg+AOFg0Tj9OhLFAlHxH3wh3ywtUZOcLrI2kZm3vvJDJ7vdN/PDjL/HqdNFgTQj2RTGVKk3b/99lu74oQIDMOYU0CwGPZAhminxx59zJJY9n9DnrgG8kSWKnO6PjvgvJBingFe93AAAJQdSURBVIdsIoSaIW3kgqAmySsrpNNPjRqzA/ofueN6gMCJP4CcIis4l4xrJhlSVsRmrABZHAKnM62M+SeQtea5kE2GkrFBtJ8DVnMCx6Y5wM6xdQeZOv5ygtPJHOSHTAKfEVHjmOhj7ol+dP7NDiBF9NGzzzxrh3VpR+QHR0dAy/loAyeT7AxNljUyRN+4yD6QR9qU4WCILu/pA4bLCATxhegm7c2/mUkCGXQ+z2P092KB0RuCFHwtuoheIB+ATCl2jcyuo7fYeQIcwPM4I0XYCuTLeT6SJ5BAZ6rBxcB5Wa4Pi6RB6HQajfeMidOQKBERK42I4cLpEgkS8RHRwSJpNKJBxvRReOYbMR5J4zFkRnQNU+b3EAQMM0LCcm6iO7IoCBlZDgTuYqXjzgUw6IzX4nggBhBGMm44TdoIgUTwiDYY0kCxcgKMLGlR+of2RaiZb0R745xxNLQ5WSUEloiFTCDkl7Qn81Vo6+yCOQtkNiA5mzZvss/4XOfnrOHl+vTjZnOcfqX/kSGGJkgpE2HwL3OsnOGArIDfEMWTEWN+hbM3EcvmWb7OubiOc07kkGEilJ6+cNLaWQWOBsfD7yB2PAf9xNw6+pehS4ZF+KM9nI1f0SGITE7AMxKRQnbJuD7Z8UmbXeW69B/yRKYK/SE6zcqGo6cDcoOjJnolg4HcoPPMeYOok+0j+CGrAHFyyiBkF8gW8sb9EiBgN5hXSB8y1yJ//nx2iTCZ4oYNGuqqq9rYoXj0A+KCse3UqZNt++yCIXr6nT8MPMNkzI+ELNPO9C2ZPUgt77E76ApOgEwd5IhsS3bAddjRnnZFFrGXAIeB3JBhhaSRlWP+D/cGISJqf+2116xcZRWQH+QbXWNpPhko5ANHxvnQ+c6dO5/QFfRh4cJAow81Nd7YZrJ42X0+FxnECJmhrSFDZMoZEmV0hGP0KbqEv2SjX3SHvkVvsfdsqvuaIcU51d2zBX6VIVXkD50geCazheyjkwRdyA866YwWMdrDs6FTBL38S2DDd/kNIFMOZziboPBscc6JEU4Vh4UxpMHofAwTHRxQOMCm8zFydCog4wFTJJVG2pbPUGoanWwQwzdEbDgMZ84CJIgImwlqpMr5w+BzPSaCcS2MFg4AZ5MTY3gpAgVg3JVogfZF2GhbxmZ5ThQpp4AQoZwQL9q5bdu2tq9g9fQFBhjhZXIcEQ4ZPlK8Tko0pw6c+RJcA4OfP19+PdXxKWt8d+3epXx589msBv0HWSaFjDPACNP39DFZLIfYZBU4G8gKRIdnczIPzJ2AvCB//DkTBnlWsh3IJn+0Q3aA7NJ3GAF+D4HHyOHIaG8yRTh5ACFENxiXdyaf5wToIfeNAYVg3nbr/262yz3RnpmHgLIL+gA5Ya4D5BY5IXUOqURW6CfIM1lFDOLZAN2HvHI+nDRGGeKF7jtRJzqAPaBtq1arqsOHDltHgtE9FzVfaDP6kCwlz04789wQXjJEkAlsGTINubn8sstztDABR8N9IzM8C+0K0BfsJc95qkPkfnhGnG3m7MLfAf2F6GBnkUvsNfdLX2FbeA7mujB06QBdJ1A9dvyYtcV33XnXaTNbLrIO+o4+oz0zZ1ZpV4f8olMEBmHhYfY4mcJLJVtHRhRZRY6wX+gjdhWddPwwWUh8FX4e30WAg91HjplC45B/bDPZVyf7dDHgbiLrwoULFy5cuHDhhUvzXbhw4cKFCxcuvHCJkQsXLly4cOHChRcuMXLhwoULFy5cuPDighKjCzmdidUhTPb6ryHzUvnzAVbSMKH0QoP+cpZyZgbHmIR6Kpj8ebrv5wRMaOXvVDBR0Fkefi5wunblOU73fIC+dpacZxfoGm16Kk537FyAe2XS5JnOzyTf07VxTnHqdZgAfaq+8x3u61zbnTP116k4W11Fvv/u3pHPzDJ1pj7PKs6kfyxWOF9y4+IvIFeZ+5PXp5Mh5Bxdu5D+NCs4VUa495zIDb85V7Y9p7hgu+ujxCwFZTb6mcDqHYpCsTKH1SasiKB2SHaWYgMaluX/LNdFqVnFxKz/fzuoPE0hN1bAABSE52QpNEscWd2U3RVTmUH9GZZTssSSFQbOKh6cN4XHWIbJSidWHrAKgVUrLCunQBn9y/GctDNlGliyT5kFVio4S7pZvkoNI2pisEqMzwBL0EePGa3FQUF29cPZ1Mfh3ik8yFJwnDfPjUJTjoDl5qwCYbWRs3ItJ6DNOB/LVZ2VFhg99IE2Z4k7euHUusFAslyVWl3cCyvysgrO6+zaz4oQp83oQ8oqUI6AVYVOyQF0heenZABtwAovVjxmB6xOZEkxq1FZZYccOCumIHaOfLLqif46W1DOgHtmdSbPwXuW79NmPC/yQF+yJQlLzVltyEqtU1dx5QTIBG3LyizIGPKPnNK2tB8ri1hdQztTT40lzNif7FybPmfJPzpBX6IPmWUcu0ibUs8LeULn0FlkiVVK2dUH7pGSDrQVJR6cWmLYTuwL9+AU0eXZKL1CCRVKeXBdVtD9nV138c8gAGNZOzrLymNWc1JGA7vLCjRWWvMvfU7hR0fXWL11LuT6bMGSe8rHsMIVecSWU2gU2eEeT7dyGLtBoVns+9FjR62e8EzozdChQ+xqvCpVLsLO+gYXLGNEfYN/Ko2P0iEIGBkMPHv/IATZBY3L71mK/NZbb9klvv92ECFQ9GuCMUoOMFbDRwzXsaPHTiw1zymoRYPBZwk0r6lL5EQr1InC6LO0EgLkLM0dP2G8JWsYUxxRTkgRz4VzQSk4N7VuuC6OlL5nmSqOgGKeEAuMMFszlC1TVsWM4lDUDweVE1DagWrBLCVt266dlTfuAbI01Dh6asVAVii4mNNsB8SD58Ho0YYOcLCcl5oltDevHVCnCj3AGLa5OnuVryFVODTq7GBcHRBdolcUQ03LFMURjFAvh+X29G9OghCKSMaa/qeOFiQBEghoM+quYPhYau/suXc2oP/fePNNS4xwCMgfbYvcstS8a9eXrC7wOaSMpcFsJ3S2W9YA7AgyAikAOAKeFZnhWpALrk0AgWxBngg0nO9nFdRjgnxQMoNAkT3oHF2kLZF5cgUsocdmEl1zb9hYdCS7gBSzxBpdo7o38gBwWlu3brV1irCp1BJDz6nb5Og8NZWof+bi7ACpZSug0LAw+x6iT0AGKab/nYKaVKmPN0EOe6tBjp19Ri8msB3cI/qA/WBfQZ4HnST4ox7bqZXYsRvIuOM/CPIpzIovoDyFj4+v8QWdL57vNgbzvMOwR4+JQDyGQXqPZOxWbQy43QXbkBh7zCin3UnYOEKPMfB2t+iXX37Zfgb4rnGkJ+2IbpTTExYW5jFG2O70Dgz7tv9OnjLZc+2119qdfP/tMEbP7gZvhM17xGPfs8P2gQMHvEdyDmNwT5zHOBGPUUjbrrR3q1atPMZgntiZHaxevdrTrHkzT69evU60d07Ab40zsa8NIfHceuutJ3bc37Bhgz2+fv16u2s61zSKY3cQN4p2YodmQ+js97ILo5SeWrVqWbnjms2aNfMYB+fp2bOnbWdkkHsyjuGkZ88J7rnnHrvbvAN2Yu/cubN9zW7UJuK2O5wbYuGpVq3aiV3bcwL0oEmTJvZcmcGO1zVq1PDEGV1x8MEHH9hd+Ldt3+Y9kj0Yp2x3XXd21Gc3eGQCvPLKKx5D/M667RzQH998843n5ptvtu2H3TCOwu5CD5BfQ7A9EydNtP3I94EhGNYGnQ1oU0NY7W757CIO6CMT9drXO3fu9Dz++OO2D7Fr2CV+w30OGjTIfierMI7FY0itfR0YGGhlHxnlnI0bN/YYEm0/ywxDiDwNGza0v80O0KNFixbZ9gK0LzrI+yeeeOJE2xry6zFBi8eQZ/usDmiLpUuXet+5yAn27Nlj7UOLFi08hujYY/QnwDZif1evWm3fY5d+/PFH+/rFF1/0mADKvr5YQAeRx1tuaW9lISEhwbNu3TqPIXL2c0OQ7O76u3fvtu8d4OOxvUOGDjmxmz764vgCbJUh/h4TuNn3FxrnPWPEtgcMJWQe5iGbM3DQQPXs1dNWt2TIC8AeiX6IjthDhsJjxtjYz2DHpNgY+SPFDBjmICIjUqL4H+wUwE7t5oaff2Gj2bMZarkUYAy9YNgUw+RfQJaEDA0pVYq9EWGcDYhOSZUDIxf2NUMfFKuDwf/555+26BupUWCcnS3ISLr0lltvsYX0cgJSxwxT7dubMRxDMU/S+M72GMAojH1NP5J1ZLiLaIoMARHzfnOPOQGFCdlageKD7OdF1ENRRyoVO5+TtaLqL21xNjg1m0b2iKrugKELZJboHLlnOIvMC/dDujknOF32jn7lLzPIGFEw8YrLr7BDKdkFESIbHJPxoto176kwzVA40SwFNNndnr3FyDCcDZAPsqIUHGToAV3AXmAneE8fMfwbGRFp+xEdYUiLrF/mPf5yAqJh5JDildgvrs1QHroBGFZyioUybJCSnKKJEybaaJpilNkBQ50UNgVkxTgvMoJssJcgbUAGlc1EHdDfOcnYokcUEnWKmdJWDIvx/rHHHrN2hQKhZLGwvQzzOMPBDOvQ7u4wWs6BTH3b/Vtb4LCokRsnm4scky1hOBhbaIJQexxdI2uErmE7Hf94seCM6Nxzz71WFrAvFKl0ZILRBQqinlpcFfuALL3+2ut2Gyr0BNlDfw4dPGSrYmN/nWkVFxrnlRiRSsehMiSBU8Pgg99+/13jx423w1w4W2eiFUMKCAHEh+Eb0tQ4D/5lbg1bP6CkGCZS6qTp2eaA4RdSwRgqgNOOio6yxzD6/2bg9BctXGSfEcLgODYMGuP/zKuAMDn7RJ0LMOyAouLkICSk8yEkzHVhWIL5KsxzwhkwTIAg//Rz9p1qZuzeu9sSAwxB5vQpz8uwCMMSOAjHKTng8+y7gwzgSGhXlJL0r9N+KDLOAjliDsXZzNs6E+hFnsW+Ns9AW2MsICoQNbaBQB+o7H0+gX4yP4AqugQT3EN2QaVpDCFDKhhHHDepf14TsFxnyC7DL6TJcwrmYfGHnGMvuAZtRj8xR435GBhTAgXHCGN/2LORYUvkKqcgOICEMIyGzDhDyQ7oR54XQ+58Fp8QryVLl9ghYeZb5BQMfb7UNWMDZXSddkb3sKeQI+b5nSswpIEeUHUdNG3W1AYqDHnQ5qfOZYEsXnHFFWcdNPx/xgCjf22vbmuHMbEJmW0ZvssZVmV4Hzi6hk0kgeD4g4sBquqT+CCAYwj/1Kkc2FOSFVSdP7VCN2TImRvIED5+xcGBgwesrrM5NUO4FwPnlRgRJRLhQHJoQDqURujTu7cdy/b387eb0BGRMwGQjBGMmOwFmQBIEk4YowYBYmIl5cX5LXv7MC8Fw0j0TRYBxwmIrj7+6GOrtGSV/s0YY9oOAgQpZN6II4jMIwE4AdoCwcvJ/IJTwbUgAhDQzCBKpN0RfufaAMJCBINDPxtcd+11Vk6IGsiWOMAAcD2HIGQ2zhAknNKpCplVIItkSXA4RGCMhZN9wAkyVwSlrmrkCvKU03lMZ4KfuWfHqGHgeAaehdfM1cHJ4vhw+pmJ4PkAJJu9mDDOEIvsgHtjbg1zHkYaAo0xw5lDWpBJHCsZE/5ljlNOAelx9gjr1auXJe/oBCQBwsV2Q8zzIYp2Jqszx5B7Q3Zw4jkFc5jIWCMjZE2QT4I6B8gMTswJzADXxv5B5AgeINnZBc/ZqHGjE9u5QPSwaUxw7fbhh5akORncswX9+P0P3+vRxx61dhf07dPXRvXYUOYzkh12gOxii85mO5n/74BwDzTEiH5GtuhLRkBmeTOB+Df0CR8HuQcEGMz/HGHsNEGNMx/sYoBsEbKNThKYEGBl3t8Mv808SbaMOhPw5yywQZYcsI0OwTiyx7zXi4HzRoxwaBhGUtlMEiyQv4CqV6tuJ4Sy6gXDARhCQCkxLjgFhjaIlhlCgChxzFkJBTAOGEDIEMMfZBcgBmSWMD585jgShlouViruXIFhHYSOtDZ7T0EOGUrA8dB2gGiO9P2prDy7QLCJsHE4RPuQHYipk0khKqhUuZIdQmNYhO8AjGROVxvxW+c8DN+RVXQILhNNcUgfftjNPitZQmRp+47t1hHR19xfZoeUHSBDnBMDhMFh9QQOgKgZ2cFB/GicIvcXnokM5hT0mYPrr7/eyjfgXzIA9Cv9CFEDdlWZ0ZOcDJGQuTiVMPKe487Z0C3HYdO3yA/6lx1wDkgKbdXeECAms9N+FU0fZj4/7ekMO+UEDGFBwCGpZJk5F+0FmDzf4b4ONgJ9+eWXbQCGTmAHINLYBrKtOQUbe2Jj2M8OgkcQRsTrAJkhkHOej+emPWnrRg0b2b51hqqyikGDBtlAhw2VyeQg7zyHE+jlM8ELbeoMuYHT9XlWwP1C+Bo3amxJGPaWYIRsFG1J1rhP3772OR1bQETP9fjcRc6AH0SmGflggju2ncUYjRs1svKDbcQesIclfQ0cXbvV+AUymGS0Lxa4Z6aqoJNORp8MMSBLSnBJogM/EhEZoZTUv0o/YLuRO4DMotMAW+v4kwrlK5zwBRca53y5Pg9FozDDnkwQDwbrnTJ1il0hwnuyEjQaCg9LptOJMknjIxiQIBgyTpqIhHQiwyxE7fyLsJBaJLpluAxH4iznJrojOoRokeJlDPZ8DIVcKCAgbFjJH9k3UvaffPKJjb7JHJFpmzVrponYH8i2U8sMCAjki6wFbUskQpaI68HoQ0NDbF+RuaN/UAj69eDBYO3YsdOSKWejy+yAeQpE3/zL+cmA0WeBCwP18EMP2+/MnDnL9muFihXsBpcQCQgxckG/47gw0tkFzgxnioKSPUImORfHIfbIGQ4Ap8YcFbJZOQHnxrHirMmG4qTRAzJStDf/kiUiMsRIsEqIfuceHn7ooWz3K+0D0eMazD+DkGFo6WOuRZSGc0d3mA9E22NgkTFnflVWwf3iSEmrk3li6Igszk0muNm2fZvmzplriR0klOHZnBJo5h7ghCGv6DZyj2OAMGMryCih/xAzyBDzNhiuJxDDTmCgnTl02QXnpW2IfiEo9Ccyy3PRrrQ1csl36E9sGbrBd4mEyYozPJtVkAl42UTekH76i+CSgOTuu+7WiJEj7DXZ3b5w4SLq0qWL/Q02kFVk3CN2Nqv6wLl4lr4muo+KjLRZAKJ07DBOihV2ELsVy5dbe4D8AoZACI4ccuoi+8CeOBsSQ6onT55s7UDjJo0zAoxFi61PRI5IAkCkIKsQVGwIukY/OXMxLzQy6ySJDOwnPolsT9euXY1dTTI2Z7L1Ja1btba6iM4iQ2RT4QFkzfAxkEPkCd7Ae+wJ7fPII4+cIIUXEud8E1nYIEaQRiDti7OkA5nHgWFCaRke2Lhpo8qULqNGjRvL3xhuh/niBDDoGG0aByaJgvI5Dorfk23C2PE574lK+Q7Xw1liqBE0hiNwbv8VkN2gXXCgCCEZlZSUZBtRNGjQyBjDnM62kSUlCDeG3Ym0SYHu279fC43RLWCcHpGMk7kjcsXoQz65H5QkJ6DvICcoBalTomIyF4wtr1+3Xj7mmbgnHDCZAeQJx8QSfu6TiaoQmZwCJ4uxQQmRO4ZhAbWScLo4PyK0nD4foK84Hxkgns8hkDwjssrzIueOM1u8OMjIc4IKFiqoZk2bZZuQOcaT7BvZNAIDSBn6QvQGKULPaHOCCAIOjhH1nTqPJCvg9wQxTj+gh4DzQ1o4J041O+Tg74CcOhkU/oW80kbcvwPsALqCzUA/nH49W0DI0UNnCAnZxDbRzhBDzCm6hI2DBHJdZ2gqq4DYIuOcmz8cIjpG+9K3yKyfv58ua33ZiUwYzoTnJUPO82a1H8kAkeGjbIZzPe6ZYdWkxCQtXbbUyqVvLl87xOFkPXHYHHfsgYuzA+2O38RvITdkhpAz+h4/BjkH2CTmHp6qaxcblKcgcEf2kH1kFEC8CXaRX7JFPCNkDlvkEDxGQrCv2BEIPufBFzCJG1J+MXDWxIhoEwOYOd2PkUKBUDo6HIeAY6ORMNB8zve5NO8B3wG853soNn+cAwXkfM73+TzzOTkXf0T+nJvvO9/l+v8V0Ab8EUnyL+3Dc/OMTloyJ6CtaE/OmRlchzamTSEhXMfpL67tfJ9r57Sd6SuuzfmB02ecn+tmBuTX+Yzf8ez0f06vDZw2dNTAeT7nmbkO5z+bazhtyDVoK+dZnWvz5wxR8Jr24Hq85n6ce8sqOC9/nIPr8XvOxTH6jHPyxzEnm8o9Od/NLmgj53o8q3Me53jmvj1bcF7OyXWcNuOZMl8XOHYAcF/n4trAuZZzbeA8u/OcXJvvcIz7ye61+X1mXXT6hn/5zAHn5w9wD/yhD3w3q3DkzWkrwDl5Pj7jerzneXgO5xl5z+dcz8XZw2lrx57RJxwDHHPamXbnj37g35zq7LlEZp3kfnidWX75HNvN8/CZ8x2+D3jPMyODmX2B0xY5AeeAVDlTdrKLsyZGzsSpzArrwoULFy5cuHBxMUBQz4gHUwVygnM+lObChQsXLly4cPFvRfZnrLpw4cKFCxcuXPxH4RIjFy5cuHDhwoULL1xi5MKFCxcuXLhw4YVLjFy4cOHChQsXLrxwiZELFy5cuHDhwoUXLjFy4cKFCxcuXLjwIkvL9dNTkxQaFimfXP62Sm9u/4zCTAmxMYqJi6c6lfIXCFDBAv/erTdcuHDhwoULFy6ylDHypCZq5qShatekvl7+tJeSvFQqPiZEn7/zgt75uq+i4xMyDp4GEZFRiv+bz124cOHChQsXLi4FZIkY5cpbWI89/ozuv7G5fv/8DfXsN84eL162mm67vLWuv72DypX8awPRk3JQSZH64deftXJLxt4pGXBrSrpw4cKFCxcuLj1kfXf91Djt23dQPh5fDR8xRGWbXatGVctoz9pl8qncRHXKFda+zcvVv+9QBc6boyXr96pZs4aaNOAHffX970pNz6Vq9ZsrKnithgwdqdFjRmnv8Vg1a1xPEwf+rKETVqtdu5bK+TaoLly4cOHChQsXZ4csT75OT09TLr/c6vrRd3qwdXW983wnrd97XHlz+xt65afU0P16/d0vVLbJ5brnlms0Z/jP+rrvJNVo0lrlqlfXlW3bqXBamPr89qeKVW+mdi1q6YM339Ss5ZuVJ4+/klJ83JngLly4cOHChYuLiixzETI56Wmp8sldVD8OGaYa+cPV5fk3tPNQlPLny6cVy+dpV0S8rm7dVLVbtdXdN7TShPEjVLh4KQUULqxq1WsqLf6oFs0M0oF9u3UsWurY6SkVL5hP7R/som+/6JJxIRcuXLhw4cKFi4uEbCdpPOnp8i1YVn/276fEHQv01W/97UlSkhMVcTREKWlp9nulK1ZQ/vx5lWa+7/GkW2aVnpYsvwKldPs9D+rpzs/rzZefU9nC7q78Lly4cOHChYtLA1kmRqmpaYpPTFRicpJ9X6VJO/3w/ceKTQhXVGy8GjRspZI+MRoyeZ79fNPm7Wp7WTs71BYbEqX9u3foQIghR3Hb9NrLbyooaLH6/PaztgZHaV3QDA0aNsv+zoULFy5cuHDh4mIhy8QoaNECTV+yRrPmzVR0QrI91vbOp/Tj+y/L3y9NxWs0108/fqIFw/vo9TfeU2TBBnrzpYdVrkx5Xd+ypn795ReVqtNU3/7QQzG7lujZ51+Rp2hd3XB5fS2ZP1NTpsxz16q5cOHChQsXLi4qslTg8Z+Qnu6Rr2/GerL0lGQlpqQpf/589v3pkJqSotQ0KW9e/4wD6elKkY/8vedw4cKFCxcuXLi4GDgnxMiFCxcuXLhw4eK/gGxPvnbhwoULFy5cuPivwiVGLly4cOHChQsXXvyniVFKUqIiIyMVExNr/42IiDB/kYqLT/R+4yzhSdPG1Uu1Ydt+7wEDj0dxsdGKio5WTHSUoqNjvB+cjDXL5mvQoBHatuug98jJCN69QUtXblBquhQdflQLFgcpMvb87TfnSUvW6mWLtH3vUe+RDBw7tE8LgpYrOSXVe+TSRcjBnQpaskpxiWc3Ohxy9ICmTVuoxBTT+NnEgZ1btHjJau+784O929do2ZotVjbOJ5LjohW4MEiHQiK9R3KOtJQELQ9aqL0Hw7xHXFxMBO/ZqiUrNtjXe3Zs1PyF65SSUWnlvwdjp9evXKZN2zJvSwXStWLRdG3be8j7/t+LhOhQLV6wQMfCM1aNXygc271Vw4YM1sLla5RyGrObFBupBYFBOhoe7T3y78B/nBjFaejPX+rqq9vo0x9+M0RkkH75/it1eaqTQs+Bn9+8YpFeeOJJjZ++1nvEwMdHB7evVKdH79KN9z6hVTv/l/jMGfKrJs5fq8Ob5uuFV97XkfA47ycZiAndq7dee0Vf9x2nuKRUDfzhQ73T7UMj9FHeb5x7LJszVc89/rTmBu30HmGT4FD99MnL+uS775VwiVvN1PgQfdztHX3w3R+KPEvbcHDjSn327U/mPCneI3+PFEMa4xMSFWqMxMevvqrvfxnh/eTcI/zwNr3a9SV9P3CqUs/z7MCJw3/TS2+8rR37TybLOcGcscPV+cnntHz96QOBSxXxCQlKSs6aHPxbEBVyQB++9IJ6/DzSvl85b7J+GTBKCZd+7JMjbF06Xy8+1UXjZyzzHvHCk65BvX/UjMV/H8gkmAAhJf1Stn8J+rPvz3r21Q+0M+TCbdYevGOFuv0yVOlJ0Xq+y8uav3yb9xMpLj5eqampGjv4F7301nvaczDE+8m/A/9pYpQ/oLjat22gmKgI3frgs3r5pRf15tvd9MAd19lVcWcHj+q3bqcrGpVWatLJGajaza9VpWJFVaJEA7VtVtd71EGyBo4Zr3KNr9MrH3ytH79+R0UK5fd+loFCJarqruvaKj0sWn75/HTdbR1UOJ9xwN7imeceHl1+091qUjWfUjI5gfyFSqjddTcrj2+aLdR5KcMvf0l1uP1m+UUnKN2Q07NB0xvu1ZJ5o1WmYB7vkb9H4Jzx+nXoRJWoXle3X1dficnnKCN5GhQrV0e3XXWZ0iPiTLx7fnHFtbcaGS6QrWzh6bmaRzfc97DqlvEYEnkxHEzOGGRC5CH1/e0Xbd593Hvkv4HCJSvp1luulF9KlOJNEPHAc+9p3IAvFJA1cf/XoW6ry3Tj5VX/d6TAx0+/DJmpro/e4T1wOqTq2w/f1/5jl7IM5FOHB+5V0YQUpSmX99j5x4LJ4xUXm1sdHn1Wowf3UsMaZe3xyMNb9PMvv+tYRLLa3Xi7ihUzfiWnen+R1oZlfRPZSwjBO9dp6OjxWha0UGt3hqhu3TrKfQZ5iDm0WYMnzdd9T72iSkVzK2b/enlqtFGNYidbgU0r5mr4xJlaGjhfwdG+KpUnXcOGDtH6TTu0ZP5UDRs3XcWr1lH54gGKOrpb/fsN0+qNazR5yhRVa3KLrrmynvdMwEcrp05QjKes7r2n7YmNcT2pKQoaP1R9Bo9Uqn9+lSxdUa3rVdCY0SO1dOlSzVu4XJVr1FZAgbzasWqxNuxL1H3336TYY8GatXCxbrzlHpUqUkhBc6dr5rxAzZw2TSpQSiXzpar3rz9rxdZDKlrAV6MH9Vfgmm2qUr2qdi1fpKGTZ6t+44Zavni2ps+YpSlTpqtQqSoqV7KIjuzeqIGDR2v5qiWaPnuumra5T62bVPHesbR/xwYtXL1BcUeCNXj4eCX5FlCFYr765aeftGTtTlWrVVupccf1c8/flJ6/uCqXLen9pbRwxhhNnBWowPnzlehfQtXKl/B+Im1bu1iDR03SvuCdGjPkTwWt2aE65lwrFkzQ2KnzVKN+I21aOl3DRk9SyUo1tXrueE0IXKfwAyv0Q8+Bik3zVeju5fqmR08diU5T84Z1dHTPZs2es1xR0Yc0dvQoxecqoNrVKyslIUJjRwzSjNnztXzdFlWvUVNLZ47R2DnLtNb07ezVO1W3URMV8PdRUkKsFswYp+FTl6tB7QqaOGKglq7fo1XzpqnfsDEqWLqSqpT76xl3rlmo17q+oIMR6apdr77igzdq0YZQY0+PqN+fw5SvQi1VLV1UIYd2avSIIZo5a7a2H4lVwzq1lCtTWBIdfkRjR41WYOBsrdqyVzXN8/vEHNPwEaO0bOkSBS5Zoxp16qpA3tzaGDRXu8PzqG3LShrS/zcdifdTheJ5NPCPX7XneJLy+CSaa43UjuD9mmv6YOzkBQookl+ThvXVwJFTVKZGHXPu4xpq5GTL7hDNGjtIo6YvVJ3GzYz85PXeERnDcE2eNVcBuT2aMGqEtu4NVf0GjY2upWrezLGaMHWGZs4PUoXqDZQ7JdToy2AtX75ci829Vq1TTwXNve7esFzDTT8vXrJAcxcu01U3PaRGtcvY8yfERmr2hJGasGClVs2ZpEVbj6qOMa5jx47SgrkztXj1dlUsF6BJo8Zo145dmjt1koYYeQgoWV55PFEaOXSYgoxszhg7QtH5zbGUoxoxdqJmTJ6go/H+ql+zsoLmzNb8wIWaNXOu0bliqlKhhBbOGqtx3Pu8RSpVrpqO71qtPv1GKzomRH1+/lkbDkWpYYMaGvzLN+rec5QKFiyg+k0bK39uP3vfYOPKhZo4dbrmzJyuuNzFVaNCKW1aFmhkd5YWL5yrw9FSrSplNHPMcM1euMLYkFmaNmOmDgfv1dIVmzRn8iit2xmsmjUqmecbqNmzZ2vpxr1q0MDI0PEdGjVmkoKWLNam/ceUK/G4Bo2Zowhjd3r//ptWbjuoWkYWCub11+qgeZpq+mj29GmK8ymkyiXya/pk0zdTFunQgc3q1auvEgqWVr0q5eRJitSwP4doxYaNmjRxtFLzVVP7a5to9NA/tHFfjCqVLqRRQ/pr3badClo4W4OGjFdA8fKqVL6UsXkHNGLoSK00/bt83WYdPXbUBJ5FVbjgX0HdioVTNXnaHPM3VXmLllNqzCH1+vl7bQ6OFOa2f59+SsldwvRBSW1dFajxxqbOnzNPRxNyqXqF4po3ZbTGzF2u47tX6Ode/ZToE2BsQuUT9hNsWROk0RMna9GC+dob7qMKRXJp5KB+Wrv9iBZNGaVB42aofM26KlMsQGGHdmmIsW2r1yw3dmS66jRvZ+x0Y++ZpF0blmjwsFFKy1fEBKGHNGTYYO00JHjC8H6avmSDmrdqrgWj++qDr3orb5ESSooN04zxY7VzX7B6Dxim/MUrKG7/BtPns43ezlVkaj5VKVtYk4YP1MKVm7VxaaD6mPbGLtauWsF7Veno/u0aMXyEVm3fqWWzxmvY2JmqXqeJju5cZmzGYJWo2kBxhzepd98/lLtENR3evFjDJwcqKmS7evXuZ/Q6RL6JR/Vjjx+0cvM+1WnQSPl94jSs7zDzLL6aMmqQdh6NUt2GDU1Qm27uebCmzQ7UrHkLVKFaHe3bsFAjJszS+hWLNWHGAuUrWkSrFs7UwkWLtWzNZjVo1lTGFJ7AoV3rNcrY/qBFi7Rxz3GjG7V1fO86fdujl3YfjFbxEoV0Wdurre1IT43V193e15+j56p48SIqW6G0Zs8PVJHcaRo9bKh2Gd2q36Ch/JVsZHaMJk03ehi4VBWr1VOxQn/ZnrSkOE2dOF6Ll6/QwsBABR88pBQVUt5ccRo3cqimTp+lTbsPqkrl8po7cYQmBa3XwU2B6tl7sHIXrqDqlcso6thejZwwXrOmTdLa3aGqV6e2cvtlLRf0L8wYpWjI7921LjhKNauWU/8vvzLG3TiiM8DH11cJcTH6/fvP9e3XX+qpzl209/jJQ1dKCVP3Lz5WYsGypulj1fvrnxUem65tgWP0Q+/+KlqxlnYvnqWfepJ69qjbm68oPL2UrmhSS4mJuU6bfTodz/XJZRxY5UoqHFBANWvVUo1KJdT7u0+1aPMxXXHlZdq7fJa6vPJlxndPU9PJzy+Xdq6YoR96DVTVhq1Upaj0QsfntXZPpKIOrlG/fn8qf5lKOrJ5mX7vO0aFihXV/q1LDXHIpc1BMzVh8lxj4FspekegXnr3ax06dkzd3n5D6YWq6/LG1RUT6/s/9+1r2m//zp2Kzl1UPgkH1emRJ7XhQITijJL+9OdYxaf6Kj4qVGtWrJBP7sLeXxleELZDH370sco0MNfbuUa//zxYyZnaKZdhBQN7faVh45epnlHsUb/30LQl61U6X7J69vpN+yI8Kpk3WX16/6a95vWhvRv1+Ref6UhcUZUplKzXOj+n5TviVa9ycX35xTeGbIQrT548Ct6+yTRUPvlGBuvZp57Xkg17NOnPXtp03E9tWjdUvx5f6OchExR6dI+6d/tA28OSFX08RAnee/PPk08he7fq917DpNx5tGvNSn38/gfGQJVXavgufflNH8VkCjwLm+PFKlQyBqCSKpYpIX9DGg4YMu5b0CjmrjX64uu+5ltJ+vTTn1SwbG1VNATl89ee1cw1fw1ZKi1Zf3z7hbYd8+jKptX1xccfat7i5erb41OtORCvK69qpY1zxunFt773/iADBQsV0u618zV44jzzyIUVsnOl+hniI/+CmjSyt3oOmGQMbgutnjNYr77xrUpUa6zwHUv0UY8/VbREEc0wRObHnn1Uo2EjQ0zG6Lc/Z3jPnAFf31yKDT2srfuiVbtGRfU2bbdi8y5tXThZw2evtUPU62YO1+vdumvWhFEaNW2DatSsrZEj+2vB6m3yxB3Se+9+oPyl66lV/cqKOGUkOE++Atq7c626f/ix9hlymxgVrl+//lRHkwqoRd3KGvnbV/px0CztWDtNXd/+Skm5ArR39VQ93/UDHTAEcM74Aeo3cIyiE0yb792pn7/+VsWqNla1Qgl6/bXXNX/REnXvM1jFqtRUgH9e7dm8W7tXTtfAacvVpk0bbV0wTl3e/MoQ52iN6t9dk4M2GWJWUb0+/1arTZBRyxjS8kVyq2rNOiqQx1tzzeCYaedPP/5VNZpcoZB9a/RRty+0e8d6fW5Ievma9VS9ZGF98fKLmrlqp+IiQ/ThG28qODRMcXFx2rNlo95/+13FpqQq6vh+/fpLH4Uk5lODmtU04Kt3NGDiXI0c2Edbj0SqTYtaWrduo+IjIzT4p27qPXaWSpcoqp8/eFW/Dpmm4G0rjTz+qNK1mql+5QC91qmLZq/cr8T9a/X9j9/oeHJ+FUwKMTr+rb3v/j9/qbmrD+rylk1UKG8BxcZ7lD9fbm1eaxzyqLnGPvnrwI7leuuTXvILqKDkvSvUo9cfIl/46/cfavmeULU1xKLf959q6oJVhqT/5ch2rwnU4HELVLthU6WG7NGzL7xqdMpPCUe36KPv+mjGLEN8t4SoUsUyiti9Tq9+0F2VGrVQ/fIF9PEbz2v6ygMKObxDX33ysfaE5VXJ3En6ytj2o2Gx3isYm2r05LcfPtGhBH9V8YvRF0bu0gsW0oqZ0/TFl9+orCFEBzct1Pc9RysxPko9vvrCEGQ/tWjRUunx5ilOySQjf+P6/2wCny0qVqyQBv/wvQaNnaSGTRtq4qBeGjFlnWoYB1u8UGFVNcFUlTJGrwx5+6b3OOOgfRU0e6y++OVP1W7SXOXz+ehdbNKu4wo9sEcfv/2Oon0LqohftL744kcdjvgr65q3QEETEAzQD78OU+mqDbVp3jj1NDapbNG8Gj2kn5ZtP64ShfJo+rghWrjhgBKNfP7c4zPNW3NcjepW0Y8fv6E/Ri1Vs2ZNNLJfT01asFr58udVxLHDCg0JMzKbVx+88rIGjV+g1TNHaMryYF1xeUutnTtRr3z4jeJN8PfbVx8qcFOw0qLDNWLQH1pkAtPGjRvryI41islkp1NCg/XWO58qV8nKalGvigb+8JF+GTpbxUqXV4WSxVWqQmVVr179RJ7K16+Aqlc19rBoAVWrWUt5/f0UHRKsbcaO1apSVr2MnVuzfZ/Wzx+vsYGbrA1ZMfFPvfzed94zZGDxtBHqNWCy6jZoqqOb5uiL34YrMd1Xw377ScGJhXVli9r65v23NGTaIgXvXK0vPvlSMb5llSs6WF98+6MSYqI0ZOAAhSb4qVnDWvr2nVc0ZPJC79n/Gf9CYuSvdrc/rVtaNdD+vbsVnXxMYRFnnjzHPm258+bXXY8+rS7PPaf77r5FBXKf8tj+Abrv8VfVrHwhBR86qNCYvYZ15jNRfW01bnKdHu5wr5667yrFh4Zq+/IpmrM0RJ263KdGLa/Vdc0rGKHI4lCDUcwqhsWXNk6pUavLVcw3WiPGBOqmW24wkWJTPdrxIa2ZPla7w2Ple4oS+5j/0lOTNXbwEBUoU1fXt26kOx95RkXTDmnB0o16udtnSo84pm3741S7QR3FhO3Rpi1HtDPUoxefbK/taxcb4d+jbZu2qlSVFrrh8sZaPv5Prd7to6eeaq8WbW5Xm/qGaZ0yZJaenqaqdRuqa5fn9MuP3+vy+vkVuC1KXbq+q3I+R0z0uUWHgoNVt/FNuqp+Oe+vDDcpUk6dnntbpX1jdCwiVEfCtiol9a92qlC1tqo3aKBb775LDxiy1fSyBsYIRqlO7brKTWTu66eq1WoqnyE7+QsEqFqdOqrZqLkefug+PXPv/SpVuqbuefAhPX7ffSpfoIji45OUatqncqMr1fG5rvrm51/UsqSPVq9erlHGeR47bBz8jv26pv3tqly+gjlfXVVqcYW6vfeOvvn8HVUslCETkIHm9RopwERR+YxBbGgIY6tWV+vRxx/RI/ferrzRhxUXn1H5HWAYypYtaZxnPZU1/ZqW6lH1Gq31+P33qMsjNykl5JgObFxlIvCl2r13j8KTfHXLbbcqT66/UpxxEfs1cdJyXX/X/Wp69b2aP2uaGhRL0dCxS3XnXTerQcOWevSJ+7R47DAdM+wS2aD8WICJYqtVriKP6aM8hsTVqF7N9ldpQ9Kq1qmpq6+7Ubffcqseuq69ildpqPvvuVv3X3udIo9FKU/BkmpYr4zaXXuP7r73Qd3TtqEiDwZ77ygDnCtv4ZK69/4O5vmfNHJQUaHhEZo0aYx27DimrUaWGjVtq2bVy6lm67Z66tH2Jkrfr8PHonXseKTmjx6kAzHF9dj97dTmhnt1WY185px/UW9f44hr1K2nKq3a6IvPPlO35+80UWuQdm43cronTG2vvUlNTb9Xr1RFV9x8k15/40X98mtPRRtnG5bkoxqNGurKm+9Qjx9/0C01pTHz1hnHvt1E7sV0983XGj3PpaLph9TtI0MKazfS7be30WgTbW7bbvRk0xYTUV+pqxrVUPW61VWmRE11uP9Rder0sJpVKaTI6ERVq1VXpUxw0bRpa+Xz/6u/5owyAUGpOrq2VSN9/G0fDe3zjZbNHKvQ6AC1u/wq3fHY42rauLQGD5qpOg3rqWWzRurY5T31+P57tW5URZdddqVee/8bvfzY/Vo2bYp2HzigfUcidO2tt6tUkcKGJ3s04o8+WrQtXs899bjqV6+h0mUa6KlOL+qtDz7Tm10e0KIpczS8bx+l5KuqO65upls6PK1qRRM0e94yNWnZSOXKNFXHJx7Qi0/frXxJUdq/f6uGD5unOx++W/XrN9ZN11ymonmSlK9gEbUy91jE2MkCRYqqTt3a5vet9NTjD+jRTg8oNCJSMSnSlhXLVKpiBdVqcqVqlyqoYuUqqFjhAt4WkVYtWqB1y9Zp85YtKly2unHErVSibFV1++Y3XV0qRO99P1a9en2hquWKatLEwYrOW1o3X91S7Z94US3K+Wr4hCk2a1vTBG4dOz6mBx59WP4F/OxcFQc+Rjc7PNxZzWtW1t6jRxUXtUGp+QPUuGFFXXlle3XocL8eaN9G6YbMb9+wXAsWbtett92lZq1a6s7rG//PkHDFGg1Vr25Vq08B5p4b1imlG9s/pLvvf0I3Nymj4wdDVK16FRULKKbGzZqrVqPLVaZUCd1h7FWP7l+oQMwBxaWUNgFXcz3U5RljV3JpxMhFhljVVfOmLfTQI0/p8UceUonkEEVH/fUcRUwwVaNBPV1h9PO+e+/RzXder0MhISpWubqKmv5PN7pdsFwllSxRwtqjKobwValbX/eY7z553wNqaojw5UY3HnrkEbWsVktRETFWVwuXrqz7O3bV65/2UKf2zbVq4TyNHzNFB3bt0wYj7/VbXKUGpn8rV6+jCk2aGbv/tL7q0V3XNKqukQP6a+r8jXqkYxdD5rw3arBsySxtOharW65vq8vb36M72zXQwEEDlKdAKdWuVEHlKtYy+lFfuU+4Kx9j/6oZH1faHG+pfLk8yle4jDo8eL+xYU+oQs2yCjM2ZMKEMdq187ixIdvUtOU1almz3EleFFsZ4Rug5i2amsCwtZGFkqpQNrfGDxytI4cPadf+47rpjjtVoYxpy3r1VbflFer0+P1GPu5TsvnvoLFDE6fM0oHdBxV8OFq33dheZQxhzCr+fcTIk6LgPRs0e+4sla/SWKUKeJTu8/fjkGQ9ShUroUJFiunBTi+pZsk8OnbksPdT4wAS47Vz6woFBi1X7TrNFJA7xQonpMrPO94BMUFIjxwNV2Ji8onMip+xlz6nkBjAEX7zP0jzztdJT1V0bKLCTTSYkJRxtsrFy5hIIb+SzzCXCKdy6EiIUfCMeUAF8uRVnXLllZicoIIl6urqVuU0uOdHWhfup8vqldRnbz8v/1I1VbZkScUbQ9+o+RV65unH9e6X3+rlzo8rxBiX5JS0E89ibf+pz2I+zGUe0se0hW/h0ipepoziYuMMuWqgB25upZ+//Vojxk3WtR1u8/4gA0S5G9YGat3mfapXq56JsFL+JxtlQlRzzQyH4+Pnay7tqzTvmDKV1O1/GQ1ph5pzGUdq25v/fHPLz48PzP84B91kXvv5mcjeHM5TIL+qVymn5KR405651e7WDnry6c76ASd6WT27959f7rzKZWSDi5z01CfdaJq5x4xsga9PmrkH7umkbys9DTL5149yOc/En3nGJNPPeYuW1P2PPKYXX3lVPX/4RjUqFLffAelpiToYH60QY9BBtZq1tW/nHiMfUYr3rrCrVrKcihTIkzHPzJ4444q0l5NdPPEU9jNf+dMW9m0uwzMzrF06H3qdvI+PCRr8MjZxzuWbntEWp8DXEFR/owM8so85B8+ekJikVq3b6NHHntAnxtm//NTtOrp/jx1GSi9aVtUqlZFpKhNkHM/QI3smdIlr2jcnAMHzM3Kci2dIMzKSml93GAfw9PMv6Ltfeqpd85p2cnuBgnns0xUsVkYVKhVXMnPhjLz4mz4EKfGJKlGqhp4wROL5t97T1199orKlyhoi8amurOyv5x+7R73Hz7fZpRbGiHLvH3f/Tq917qD05FR7H7lNdAuMptt/yU6jc84TOAiLiVBU5BEjGeZ+ipRU5SK+2n44SqHxcYakYt7zqVrhEvIxspduzH1u/zzyM+0IPOZXeXLnM89rWsRcV/75dcs99+sZE7T99ON3urJZbd32cBe91ukudX/reb3V7WvFckq/vEbfM/qqUqXKypser/2mfVNSkux95PL3VwND0lOMLKXZnQgIITPg6+OnqOg445BibZuBXL5/7VbgY77PK56S582d29+2AOelvxDnOzs8pOmTJ2nmzInyr9RId9x4g/n0LyQlJqpszebq+PTTeuv9j9Tt5adVKE9G/9SvY/QtLFhLV+yw340MC1Fc5F+rlKrWqWH0EftgWj6Xn2krc1HzP19z4ZM005OqTWuCFLhoueoaslAwX56MeXaZ5NjXyDFyxjDtsXh0g6fCXnCmTOfywmMFMuM4l/XPlXEeP/M7dD3d2Op0Y/vSrNc2vsbISYF8Gbs6hJtgLKPPzRufIqpRqIjSk5j8nG5thtUZ04oZNsP+JBN8ja3yMhAjDL7efuFu8VfA3pn5P+TvhB5yzNff2GS+Y77NMYTJvKS9cnnP2aBuFXPvKUpISFGDy2/QUx2f1mdff6vn7r/J3J4Jroxt8PPaqXbt79NH73TV1MHf6Iku7xsiEW6Pg1hkPTTMPBF3ZshktermtxkP4/GkWfk+Fcg47Zbx2ty76VPunYfBDtEPiUnJuuyKdlYPP//hB3V54taTVtm2vbqt8kauMcHFKK1ed1SP3HWrShZIVYpPQbW/+yF17NRFP/7wndo0qKxk88Nc5hrAYzqR66Uam+GfO7/uvLuDOj7TWd//2l1XNq2ZIS9ZQEYP/IuQFHZMo34frJqt2qtmBX+FRqVr+96d2r9+uSZMnn5iSMQBBs/8zzR6Rsf65S2mVbNHa8HKv4YyQndv0cj+E3StafDCeeIUHp2m9ds3GaOD8c1oSkQgzQhabRNdekwU+kffqfb4odBERRjhORUxRpOiUv5KA58AWmRuKDwiXmVNtN+yYQX1GzzBfrTv8B4VrdtYFYsFmPtNMyKXIYBEA8zw9zPG75Yb2yho8UJt2B+ppKRYhRphq1+/of1eh3vu1Mh+/VS39S1695VnNHPGfJU2gpw3Tx5VrV1Vk/p9p18GjjfR3Qz1+eNPVTbRenjoFg0fGWR+najg0CRFxZ7yLOZe4+MSrdKF7tykYyEpuqGVCc8NHn7sEYXtCtL2iCJqU/uvbBHYu2KRiQxX6O6H7zfGItK0aaLWMczlBX3CczkKl2bIBUbHv3BhO1Ev7OhxrV2/UhFRkcaZJZjPTJua++Bfvoex8kHtsBwJsUY5Ug1BNi8TDGk1MhB2MFh7on1MtNjGRHJ59Pk7L2v+knWaMba/Rs1aYYyCr2ISzPm8cpEZqSi8+aP1zS1a2bGvzVfTzN9JjtJ8kJKQpIhjx3TI3HNiIqQ143OeLc0Y/HLG8HtidumFV7tp/fpN+v7rr7Qj+Jj9DshXpJKqFIzV7z9117Y9uzWk19cKTSui5nVLq++gifY7uw7uUpnGLVUuX26lGKODbKC8+QoHKPRoqCKjo7Vi9QolGbmLN0Qb4hfn3dMwY+K8955p89gMp2R4yAkiilFy+iIzuJb9jvkfBJCXDWtX08DfvtSQ8bMVtHCOfvj5Dy2YMdYQ51pq16S2jhwL1e7dm1WhWjnt2rFSk2ZtNNeKs/IVYxzWSTAnjDGkxvZDQDlVKZWut55/SYtWr9eYP37QtGVbrQM5GhIjFhuGHd2rPP4BqlmtmpH/RMWZIAWUrFZH4dvn6u13PteWLRvU+/tvtGrtaq3ecUxf9h2pT19+TMvmLzSkqorRkW81YNR0LQmaq29+HmwcDY7FBAg2wDKyBSWgwz0+ik1M0OFt63Uo8q/x0yuvbKF1gZP069BJ2rRysb78sb9aNm6u6B1LtGwX5DZGu0MjdcV1bSwhSko1ZMU4dQtzzqTUBKWatgyoUE7FA5L0+utva+nKjfqz1y+av3S1Zsydrtte+FKzJ/bW3g3zdCgmVblijynUBCRg+659qnvZFXrg3vZav3a5gjaZa6Yn6WBinBo2aym/VKMD1lvTpxl2q3TpUqpaNbd+7zncHg8xJOlYZJSSzSPTvylGj3hk+oOuoCXMT+2/9Dl3X6pEcYWGxerZV95U63rl+cYJVK5RS0HT/jBkdrA2r1ms3/oN0rGwSI0dO0LFGrfX1689oPffeFm7jyerzZXXKmLXGs1Zx/L5dCMrh3TdFVcbGUkx9+o14OaiyK29Dy+S4o6rT8+JanXdnSqYJ1lx0RHat++gjsXwvYxvEqOwWrNYmSryjdquKTMCTSydqsPHjxtC/7+retON3eD5QKr5oXPFDH0wt2H/L0YHjhzVod27LJmIMfoOrmzYSMFr5mhdsLGZKceNvUnU1ddfZWUZvaFBM2zGSRbDwto+74XTnWDZ3xDm3Ll17OBRHd+zTXv371NMUpKRFWODjS6neOds8F3bV6bPPEYHKEvDgpNE85wpVh8SFLh6j5q1vUmN6lVS/+/f15AJC7R8/gT1HjbZ3I/5hvleipeJLFi2TJVb3abJE8eqQOoBE8zussdBvfotVTAhxM4DBDt27tbVLa60r9PMTcSGRp1ovxMw76PjYnRw6zoFRyXb+7XPav6w86BBzcr645fPNHLyPAXOm6Effx+lzAM5CQkm+C5RTjHR8Wp67V16osONJqAooqJGXz5+5w2tWLtZ44f11rSgzYZ0GfrplXcaPNW0Z9GixRTgE6lu3bpp5YYt6t9/gOYuXJtlwvOvm3ztl8dfx4I3q9dvv+lwUgHFH9qvPMXrKP+xrfp19DTdeNf9KpxB+o0R3a+evf/Q+m37tG/HRq1ctkRD+v+qRUYhn3z+FZUomMEy8+Xz1+b1i9XbKHOCv1H+3XuVO09BLVuzWodDw1WpZAmNmjRFu/YfUbs7HlGjyrnVo0cPBS1dqsMhB5SSu6CaNm2iEkUKIeVaOnWUBk+cpp1Hj5motZpl7xbGac0eOVzjZgbq0JE41W7SRLdcc4VmjfxdC9du0UYjkJ1ff0fVi/saJ/mzNu0/pFo1GmnL0kmaOnuNMaQ19cCD9ytie5BGTpmpdYZ81LrCRNcP32jHePOaKCXZN7/uMhF3xdIlFW+M8E23dVAZ0yBVqtVQ7LHt6tXLXGvVFt376NO67bbbjATvMo7tVy1bvkJHGZLMX1wtmjU+MamSaGfpvGmaumi1nXh5w4Oddde1La1iFixRRXtXzdJNT7+phlVK2+87yJ/PV/PnTNTQ8TPlm7ewDmzdoyZX3qIGNTIM6qolMzV84AjF+pZQIf8wTRw+UrHJhXTdTdcrdMk8fd+/vxKTDRkzxpVgYNvm9dqycpPyla2m3duWaMHCJcqTt5Tiw/dq8pxpSvUprWvatdSSqSO1ePUqLVpg7vXJzrq3XQs1bNxAs0cP1Z/Dh+lwQoC6PveIZk0YqsXzApU7oJquaFkvw9AYpBnnMnLEYAWtWqvcBYtr88o5WrLtgCpXLKPFU6Zq0dZdqtuglepU9xJBE+nFHtyh3oOGG+dTVEuDlmqbIWXlS5XUtJlztHHHbjVuc6vat22ooT/9oKETp6pUvavV6b6b5e+dCOjrl1f16lTRqD8H6M8hIww/aKWurz6pRpDZIb9q+aZt2rz9oF55732VNMT9t569tP3AUTVqfpVqlCik8f16atisRcbJFtPBw0eV7puqZbNm63iUj8qWD9DEyaMUfChaNSqU0cwF07V99wEZfqV5ixfpSGy8KgT4acL46doTFqFWl7dTqWIF7X3NN2Rn0pgZSsxfwljAPZo0dqJSC5RVp2ee0vEtC/Xz7wO0ZstuPf9ONxXxS9affX7Vhq2HVCq/IUFxufTsCy+bsHqrfuz5u1asWm3u54j8i5RR62aNlD9vHsNnIzRy6J9aEbREhUrWVovGddSsUU3NmjhCAwYOVrhPSb3xUkcFb16qX/8cY4x+vObMmq+7nn1LNQrHqn/fvtq/77iatr5a1Y1jrlzcR/3+6KvR4yapXrt79fD1dfX9l19q6YbdhlzH68rrb9IzTz2qIxsX6Off+huDuUOdX31Fh7cEacTUOca5+CstIlxjZ5pnTiuka69prWVzR2naihDdc097BRg7AcpUr6uUkN1GR3/RPKNL9zzaVXfeeY384w/q9z9HasdGo6sVG6pLx7s1ZfCvmrB4owqUraTWdStoaN/fNGvtTpWqWlPNGjZQLdPHc0wb9Bs2UkkBVfVCx4e1eMZADRw1TTGhR1WmWXvd0bK2naA9f9VOHd2/UQdic+ut119S45bNlXxoo4aNm6j1KxaqVIOb1OWhK/X7H/21aus2lShWWsuXrlCQsWNV6rfVXbe00tA/emr8tNnaG7xbscZhlShWRAsmT9TmA4dVtXo1TR0/xDi/EJWtUlmzJg3Txk0H1bB5G+2YOUYzlq7SpvXrNHn8GE1ftEmtr75ORQtk2NAqtWorLfyQscs9NcM4oDsfekq5Y7fpwx6DdH+nl9W8ah71HzhEOw+F66Enn1Jx3xD16jtc61evUeFal+uFx27RxFH9tXDuGuUpV1mHtwZqzrQgFataX5c1rmWv4eeXW7tXT1O/ERPkk6+I9h8MVVJEovbs2agdR0JUpWxRzZgwVWv2Buu6m+9To1oB+sXo3AKj50fiYxSZlGzku62KF8rI+GxaNl1//DFW6QVLqIiOGN2cp6TcfiqcYuRy0hxFmgCtbbtrtWdbkMZMWaxiRYsYJz7LTgRu0eIaXXHtZUo+ulV9h0/UFkNQKzW5Vk/e00YjBvyiuWv3qGLl0lo/e4ZmrtmiqjUbqmn9ava6+3et159GdlnBXqZcQc2bNEq79kSrzQ3Xq2jYYf3U80dtPRBmSFu6CbQSdPTYQa2Zv0jJBcopJZ4hoilKSCmg0rmTNHzyGIVHeXT1tdfo8KYlmrVgjpazqKD1TXr1qbvVoFEj7Vy2QL/3+0Mrth7XCy+9aMj2fE0ZP0HRiQV0a/s2Wrt4orr3HGT0K1H5ipZVh4cfUeG8GdmkIoZgVirhY9qxj7Zu26EwTzF1e/sFRQWvVc8Bw+0Cj2JFKqhhg2onbKiPIenTpwxT0PYkFcsTqbmTZyupQCmlRe4wsjNZnsIV9UynJxW8bq5+6f2nNuwM1nNvvKXKpf6ao7p53QoNGjhMW3Zt02ITfA0eMli1mrfTXe3basKgfsaWj1Bywep6+pFbNH5oX61avksBlato2/Lpxq6vVe0rrlOHm1pr3PA/NWDwCOUtVdUO0RbKmyGv/wjPvxGGkicmJpl/0j2GbXsMCzX/pnhiYuO8Xzg97PfN32mRnuZJMOc0L8z5DP88w9cykO6Ji4n2xMQleBLiE898ziwiLSXJExIa5klMSvEe+QeYe42MiPBExcR7D3hhjqeZsMfejXmdmpKS8foE0j2RURGeWHPPJ2DuPSY6yhOXkGSeJcF78BSY70SER3ji4mkfTp3mOXb0qGfvxoWeZ59+3xMRffrfpaelmn5K5pUn1YQ9WUWqeYaoqGj7b3xiFtvEgZGDqIhwj4mwvAcykJKc7Akzz5BiznnOYdonNdW09T+IQXJivCciMtpjxPW0SEzI+DzzeVKTE41shHuSks9833GxsaZvEu2z/d33ziXo26jICCOzf7VzYmKiJ9XcPPp4or+NHEZHRZp+TD6zfJ2C1FTTV0a+k73nGPDNK56nXu/miTHPFheXSXZPg/j4WCOPMX/JvbmfqMhIT0zMX7bBufeETPd+JqBPJgL1vssE81wxPNcpchYdGe6JjI49Re/+Huh/eHik6T/nV8a+8RxGFkDsvi2em665yzNt6Ubz/HGZvgfSbftGRv+97XOQnBhnbECM1csUYx+yguiDGz0d2j/k2RSW0X9RR/d4Pnq+s2f2ioP2fWZEmXv5JztsYfol1vRBZJRpq2w0FrY5w/ajG8n/2M6xscZOm/tJTk4y/ZiNC52E9L+1G5HhYf9ri88CyGtyirGdxh+gS1kGbRod6Yk+5V6w1+HG9mX4t9MjydiecGM3z3S5BOPvwiOistyG2MO0f+jYdNOmkeaaickny2Facqznsw9e9fw5err3SKrn5y/e8vQaM9u+S0pKtLY8K22TnJjgiTC2JLtd724i6yLbSIwLV+f7b9SGw/n01hfd9dAtl3k/ceHi3CIh4ojeef4xBR0vrGGD/1StcoW8n/z/warZ4/To46/qobc+UbdXn7zg8x/Cdi7SHY++ouKV66he9QpKSkxR/TrN9WCnR1QwI7HgwsU5Q3J8lN5/6Rkt3haqlq1aKF+edPkWqaBnOz6mqqX/mpt5PuESIxfZhom4tXXDKiX4FjEGkpVjrnV0cX7A8Nm+vXuVkOarylWqqnDBrK8s+a8gPPSYDh48rAKFS6hq1YoXYWKoR8H7duvAoaN2vleu3PlUt37D/13d68LFOUJ06HHt2L1HSakeI2+5VLlqTZUtUdT76fmHS4xcuHDhwoULFy68+FdS/mMHdmrTlj12BcI/wZOeZpeSUlwtMTHJTmbjdVJSEvOrvN/KPoL3MCF21/+sNjjXSI6P1PrVqxRBMZHzhJS4KAXNm61ZcwMVl5yFRv0HHDm0VytXU7fIe+A84/C+LVq7ccv/9MXBXVu1edd+u+z47xAXE6ElQasUHZe9TdZSk+LshP7wTHWN/g5JcZFat3adomKTlJxg2jxopaLtWuxzB48nXcms1rLynmhkP0PeE428J5vXsXHxVvYT2MsoKwrkwoULF//P8K8jRhFHd+rll57T1/0mnXY331OxetY4dez0jH7s8ZUee/AOPf7U6+rx1Wd64PEuWrk10674pyAlmaXhp79AyK5VevOFjvp9yBi7pPO8wZOgP3p9r6dffl87j52fzQHTkmPtioVVm/foxy8/ULcfhno/yTlWzB6nj779XVFJF8bxLpw2Ql/83PukZaPH967SS88/pT/GzP5H8hqyd4vefudD7TySjZ3fUxM0a1QfPfnkY9p5/DRlGf4HKfrju6/UtWs3BR+LU/SRrXrjjfe17ci53XV69+qFeu6Zp9W9+9d67qkOuu+h59Tj22/1xBPPaPiU+Zo2+DvdeOOteq/b13r28Xv1zEsfaF/Yhdt40oULFy4udfzriFHRMjV1VaP6SgiPtXVr/gnBx+P07Kvv6I2XnpV/SrQ8ZRvpnY8+1t1tr1FqpiJjJyNV3bu9q5BMVVczo2T1hmrTsqHCo86tU/sf+OTTrbfdpFxh0catnp+uOnJgj2ZNmKbW192nP/r00j3Xt/B+knPc+khXjRnwtYoXuDDidf+zH2jgT9/YAm0OSlVtqlatGissIubEMtIzoUqjKzRz5hg1rpqxh1eW4JdPbdu1U4E8fkrM0u7r/rr57ttVICVECYlpKlHtMs2ZPV7Nq5/bcfODx2J052PP6t03X1WFgFyK8C+jF954Vx3vf0AByqurr7lK24OP69oOnfTVJ+9qzqCf9f4XJ2814sKFCxf/n/Gv3ER26/JAbdx9XLFhxqnPXagCpcqrXIki3k9PRvkq1VSjSkX5+6dr8tjRylW+lTrc0EK16tdUxYqlFXpwj6bOnKNlS4J0JDJFtaqV04Q/f1G3b/qrTNWaKleqiLauWaIFi1aY7yxR3qKlVKpYcW1ZMVd7Yvx0T/vrTzje44d2a/q0Gdq6/4BWB07XtHnLVK1GPR3ZvUYTp0xX8Yo1FXN4q8aOn6g8xSrqyPbVmha4WnERezRw2CgdDouTb9Jx/dG3n7YeCFHN2rWVxxOj0QPGKG/Jwlo4Y7yORKeoeq3q8vPxaNHsiZobuETLV69TuUqVtW7hXC1ds1mrgwK11Bxr0LTZic0AUxKitGDmDK1YvkpBy9arXPUaKuQbr6GD+2vC1JUqVba0Wlx1lepUydgh2ZOeosCZU7Rw6QotXblOxctVVXLkIU0YPVrhKcmaNmmc5i1cpbJlC2rCyGGaPHuxbeuAfNL0yeO151icAvzN62nTtHPvAa1YNMe0x1KVrlLT1hEJP7xPk6bN1MplS7X3SJTq1Kp6EoFJZMPGaZO0cuUqrVm7VeWrVleBvP4K3rVJ02fN1tIli5XgV0Ql8ns0Z/oUbTsUqXo1q5rfhWrK+PHasmWzpkwcr8JVmumWa1pp9aJpClqxTgsXLVKFOo1VyLvrcHpaijasWqQpc1epauXyWhU4W8s37NDO9as1Yuw4peUtomoVyiglMU5zp0/VYiMnm3fuVaVa9QzRiNOfQ0fp2rseVoTp40lTg1S1Vg3t27JaYyfOV9kqVRWQP4/WLVmoeYuXKmjxfG3acVi33nWPDu5cqWVrNqlQoWJav2KBFixeq0MHtmjYsLHyKV5eVUoXU1JshKZNmao169dr85btOn7smHLlK6HCmXb9P7h7m2bMmK2FS5YqId1PLS9rpdrVqihP3twKmj1ZwSml9WSHG1SjZmXVqFVJaQmh6jN0ph56/Fk1qFNdK6b1VbingO7tcPeJSskuXLhw8f8Z/8o5RpT/3r56scKjYhQ0aYge6fiyDmbabDAzAgoHKBce10P1UY/9F+TPn1++KRH64M3XdDgpn8oX91O3F1/U4CkrVKZIPuX1y6OCRQordOcGvfpOd+UtUkTrF07QD/1G29+fNgthzj2w15f66KvfleyTT+P7/qDfR0xVfk+0enz9iRZvPa68njj1/vFLzV67z+6y/N67r2vSgs3yS4/Tu1076fdBM5U/r5++/+ITTQvaoHz5DIk4ckA7tmxS1OFdevaJJzV65iptmD9OY+dvUo3qFTVxwK96+4sftWvzWr309LNaunaNVhlCQVXbDHg0pn9vDZsepAqVK2rllIF68+PvFZ3oowL58ypPntzKV7igeaa/Bp1Cti/Wmx98ZIhFLc0e8LMhbvOUR/H644+eer/7YHNffpo09Cc90+VTpfnl1bIZw/Rlr5G2AmvQvCn6ptcYxcanataQH/VZ958Ul+anqQN/1499xpmzp+iTt1/VnihfVSidX1+/9bp6DZ+fcWEvFk8Zpe97jVeBIsXVt3d3zVi6QYo9qG5vfiqfwuW0Y/U8Pffcy4rzyat54waq15+Dza+S9VuP7zR39T6VKlFMiccNkfD31+Fda9S9+48qU62WNi+eqkORfw0dsV/XoR0b9MXH3+l4VJx2rF2uV597Xhv3HFFY8Ca9+/5XOhqVron9ftDkRdtUvUJBdXv7FQ2fslh27wGDXP65FRO8W1+88572hcUoOeyIenzQTdsOHNPezQv1VfdfladQWeX1STLkO+NnRw9s1nsf/6B9oanasXC83vvgPW0/HK29q+brjTczNv0c/uvXGjx5pSqWKqSBv3yqH4fOUEr6XyobHXpAX3/ypSLTCylh1wJ17vqmjiTm9m5tgazb/1mR98+dx26yy7y6lJREhYYe0dIZfypwW4QaXnGTMkreuXDhwoWLfyUxSktNVp3L2uvt999Xr14/ymfvZm3efdD7adaxfNpoLd8er2cev0Pt731abZqW0qB+o1S/eSOVKlZG17Zro9r1G6hTl6dVpURBRcXEadeBECWmG2J0GmZUqkIN1W3SQJe1u1FPd3pWd917g3YHH1TF+o1UslhRUfW9ZPXaKlu6lHL5+qlWo6aqVqe27unwqN58rosaVK6rltfcqle6dlWrKlUVdjzCOLI0BZSqrEefe1Pf/DZAHS6roaA5czRqyCjt2XlAu/ceUJ0mLVW0WGk1a9VUTRrU1ktvfqU+v/2iwt5V9PGRxzR0wli1uuZmXXV1W3V94wUtmTRYW4/G6Nq2V6ucedZrbrpdxQtlVLsGBUpV11Mdu6pc/nQlp8Vp5/71KlKhnipVLKur27VXx44v6J7r2qlI2cZ6plNnPXT7zQrfHyI//wK6unULFUlPU6kqVdS0bm01bHKdunR+Vs91uFoRhw5py5KpmrPiqJ55qoNuuP0R3dKutgb80uekSdJkdbp0flh50xJ1PCJewYdDtWTiBO3xlNYD7a9Rty9/Vs9vP1TJQgXVvFULlSgWoP3b1mv03MW694EOuuram3Xjne3tRONcfv5atWyNRo+dqadf6qaapU+uhdOiUXOVK1NGufIGqMXlzdSoTl11fPYlvfrSiyqecEzHD+3RULaMufcJXXXDgxoxdLhuvaKRlJpmiYavfx7Vq11HJQuXMILhp5o1aqpCiVJ2qfmo3n0UUK257r2tre69/1HVLp6qVEOar2rbRmV9PCpYsqQub1Ff5co1VefnOun9lx9RytGMzVwXr96gQtUamWe5VfXq1VS+EjVUrdxfG3duDpqmLRFpeuzR29T1Q0PCu3+isgF/r9LsR5eSHKvFgVMVtC5YPwwcr49ffMj7qQsXLly4+FcSIyLgfAUK2YnPlSqUVbXKRRTv3TPpH5GJ0Ow6cFjJyWknHHKrGnWUlhythCT2sUk3fs9HqebfbeuDNDloo2o1bqDUpNSMMPxM8Mml/Pkyhjp8/f3FxrPs38JEbh9SVx5ugE0FM3ZI9zNONX/eXGKH51z++ZWPvRpgXWwAajcHNN/JnVt58hLT+6t1sxpKTo02JC1Jra6/Ux2ffFo/9Pxd7z93v/lqsvKbdmHTSsiAg+TkBB05sN90dkZ3Fy1dUSXKFFZcfII5f4rdZ4bVTJnBSr6tGxdo9aZgVS1f3txfxjyadPMshdn6xMDPPEtAQMb2EWxEeWKLC9M+JwTLPEq+vBl7tLAZLYRwb/BRJSSyQac9rJY16hq2G6PMPZiSlKCJU6fqQLxUtWJp+aT46HD4ccUmhtv+CiheTq0bVFW8uRa7brPxYkxUhKJCwpTb++yeXH7ymIuUqdxA3/f4UuvnDNTt93fSmg1/7ZOXgYw+sa980pQ3Xz67EaGPb7pys220IYb7EhIUER1qv9Os1WXySyXrlPEbwEbGiIUlzPyf+UtLS9Ou3YfNObzyYA7nthswmv43X7a81f7ItKWfn6ClbNDIf7T2XTddqS0Lhitw7mxFRedThxsztmJxEB9xQLEJsYoxDedXqLSaN2mouNC/n/dGP+crUFx33vuU3ninm+6/pZ0KePvNhQsXLlxk8l//JrDRX1Jiqhgw2LZ9t2ILVFDtymW1Z+0KLV+7+fS8xThOu2sxOwx6cfXllyk2Zr8mz8nY3HTP8UOq1aKN8rGrtideYVFxGjOon6YFbtWLTz4kT1yk0tITtHPPfktYGNLL7KgATi1jy1EvcJJ5jKP1z63I0CiFH9qrQ0cOKYFduI2nTISYOcumze3ZDjG/8U0198BmkOZtUmq60lKgA0lauvGgGrS+VnWqlVbf7z7UzEXrtWnFHP02eKIhGn7mvpNN+2TOvUiFAoqrUbVqmjc/Y7gqIvyYigWUMoSngv0mm6CmnkIsl44ZoiXrovT0E/cp1RCryJgYRRhH7O/vp/gkL0ni2bw7QRsmYUhjkr1fjnvMe7sLvTng3TjaNkWaee4rWreQb/pxjZ66wh7ffWSfqre4OtNwTqqmj/1TifmrqsPNbRUWHqWDh/aoao3K2r92vn4bOUMH92xT9+4/KN48gI+VCKlI8ZLK54nRpDmLzbt0JcaEKyE+Sft2b9RRv+qaOy9Q7RoGaOL0ufYqJ8Hcox/3nZ7L9l/GvWfcuF/BcqpRzKPBvb7TnsPHNGt0H01ZvNV8kLHbNWQvXwF/Q6iSDGFN0IFDB3Qw9LiRD181aFhB82bPVnBoounPBB2LTrRElc1v00zDQOo4B5fjXwgy7cWV8+bxU4VSpbVt51E98MxLur1tE3P0L1RucJV2rQhSvz8m6Mj+7frt1190JPYvgmt34M7Y9fYE0rk2mzp6N6R0EHfskGbNmKWoxJOPu3DhwsX/N/wrJ18nx0Vr/JC+2rR7nwKXrNVDL3RVu6a11Lfba5qyNUQ339zupImk7JQ+ffQQjRw1WQdDjqtWg8aqVqG0ilSsoXzxRzRg4J/at2u9DsYHqJs5B5vLTp44TFMDt6jNVVdq7ZLpGjNpgfxz+2rzmlWqXb+Gpowaq63Bx9Xk8mtVoWSAvc6+nev0+08/aF+IjypVKqzJw/pp665o3XDrzdLOzfqh56/aceCowkLCDCFK1ZHDexU4ZZZUtIqSo/do2IhRSkwvrNK5E9Rv2J8KjfHTtde31abAqVqwbIlWLFiggvWv1OvPPqiGjepo6ZTR+rlvP0Ng9qljpye1ZuYwDZ25TAEVqunq5g3sPQFfvzyqW6eyhv/ZRys27VHg4jW6/+kXdFX9Evr5++81fclK4zzz6LLLmymvd1Kyb3qMBvzxu+as3a6k2Hjt2XVMRYsV0axpU3T0eKIqVy6pwYMHGKIQZkhaNU0zRGrp9u2qU6u+Zo4aqCXb9qh8qQCNHj9O+4+Fq2b50ho4coS27jmqG+97TA3L5lbv3j21f99W7TguffTR2ypeyKlq7KvjRw+q3289tXbDHuVLC9HOkAQ98dyL8o/arx++7q5pC5aq7e2d1KJKPtPmPylo/X7deOe9alQ2v35kntGi5Vpt7iEuOkblyxRW9+++VkJqLkVFhuvuJzqretliGZfypGvKuKEaPXmK/IqU08GNszU+cJ0hYRW1ae50jQpcpup1W+uBe9pp6O+91H/wUEUkldTrrzyhxZNGaPDoqfItUELXXXettqxdoJ9/+V37gw8rMiFGKabdH37iMS2eMkx/DBqpDRvXmfYKVolKVbVr0SyNX7hCZUsV1sLFgVqxcbMqVKikBfPmat6yVarb8mrtXrNI46bO1q49WzVvznRNmT5H7drfqkJ5M6S7aMkK8gnbpZ97/axxkxep9U136o5rMsjTkjmT9Ge/4dp1MFgVqtdWw5qVdfjgbv3W/XsFrVqvZF9/XdaqqQp6s5u7F81Q1/c+VuvbH1X5In9N7nbhwoWL/2/411a+TjTkKCwyRoUKF1WAdyf4uOgoJZsov2jh/91PKSU5mREtA48dJvH3TpzFMYaFhigxRSpZsoRy+2ccT05KUFJKugoVLKCEuBjzucdOVE4252FCdGpqRmG+XH5+8suV8Zv0tDSl2OM+NjvF/CCidSYA5/Kk6vCxEBUKKGKHo/Lk9jf34QNrk8fH1w6zpNuxJR+7o72TRWLSrNKSFRoaak6UV8VLFssYgjFITU7UkaMhKmjaoGjhguYZk2zRSx9zbc5/KhJiTZtFRKtQkWIqzHwi8+xJLDW32REpt/mNzZR4EWvaM9WTSwXy+tm2yJs3d8Y9Mizom8u8tuka5fLxUxrPStua4x7TDowYkv3gWQHtkfF8DBuZ9jDPGGHaPS4pXSVMu+fNffKuxwz5REdGKlfuvMrt56tkc3ra3zSyQo4fMw2TTyWLF7XPkGKegaswfOjHeQ35TUzLmFhOXzOUR0HDmNhYFShU5IS8OEhJMb8390YWh3wft5yL/iOzYj7PZfrX3/RzXEyUwqNiVbJUGUsg7e9MW3Dt3Llz28za8ZAIFS7MLtEMk+ZW3jy5lZQYp5DQSBUsFGDOn64CBQpYWeF6Gf1OF3AWX9u2vkY248KD9f5HX+rR515X83pVFRMWok/ff003d/1BNzfy7uxvQAHTUPO86bny2AnnTu+lmntLMye2702fMCGb9qetfMxF0Xp/ZNDb32QnQ8KirA74clMuXLhw8f8U7pYgLlxcgji4Y62ee/ZF5StfR80a1jE0K0WJuQvr9eefVWFvxsiFCxcuXJx7uMTIhYtLEWmp2rNtszZs3y0fP3/lzpdfLVtfphIBf61Kc+HChQsX5x4uMXLhwoULFy5cuPAiY9nN/wOkpqbY+TTeKS//C3OceRlsNHsmRIcd1q49Byhfc9ZITYrVju1bFcOyqguEsKP7tH3XXju/5HRgvsqeXbt0JDTSe+TsEHX0gBbMn689wUe8R05GSkK8tmzbqdiE7G3e+v8JcRGHtHnrjsyLKbMAjyIPH9DRkCjv+zMjPjZCGzZtVwLFuTIhNTFO27cY+TwHmwpnFcwrYy5gSkqqnQ+VnJyklLQ0paG7SYl2vhpzpJg7lfGDNIUcO6iwqKzsVXdpIy05Udu371BkbILSU8zrrRsUGXNu9rCLDDmqHTv2et9lwJOWYovGRmZxA+TTgnmTB/Zq5+59SjICmhQba86bqkP79ujgkRDvl/7C0YN7tWff6W1BVsG8RjYCt7Jg7HVWNkKO/oetmyhNkpCQ6H33F6KiI7Rz/yE7n9SRR+qiJSU581UvLo4f3q+dew55313aOHokWLsPHPa++3vEREXZuZ1/IV2hRw4amQq18zkdpJt+ObBntyKiT9Z/T3K89u7Ze2JT8PioozpwNBv7YBr8K1el5QShR/brp+6f6adefVXnqltVJiCjtg6IPr5bH77/pkZNX6aadeqodPH/3V4kPiJYb732sqavC9WN17VR3pPnCmcP6fHq/fO3+rLncLW+5g6VLfLXvZxPTB/+q34cMVv33H5LRjXwU7Bszni99u7HKlypvhrVrOQ9mgGcFv4oqxNzj+7frB59xij66A599F1ftbiincoWz5gUbx2bMXCj+/fQhz8O1tXtrlHJohmfnW9kTBjPWGZ/KYAJ3iDzpPfM2LpwlF79sq9uubeDCmRR5ratWai+Y5eqlV119veytXvdfHV5rbuuvOk2lSqc2xoeT2K0hvzxkz79/g+1ue0hlTyLPe/spHbzx4KAv0N8+FH1+PIjTQ9criVzxuq3fqO0y5D4yaOHafm+cEVsX6aPP/9am3fu0aQxwzR94Wq1vvIqRR7YoD6jZql2vfoq9A/PeiljitHNd7/qpaaXt1VRvzh1famLytS5TNUrlPJ+42QQxLBogv/+DmEHduvTt9/S9GX7dM+d12UcTEvWwsmD9fwrb6jpTR1UqXhGLbLsYP/WFfrkky+1dfcRxUQc0fjxozUxcJ3K+Mbp7dfeV1zeUrq8eV3vt42cbVmlV157XTsj8+qmNk29R7OP1JQkTR45UJ989qXWbtmpWZPGaPzMhWrYvJWKFHBWtWbg8E5jgz5/XxOW79Vt113pPXoyguZM0ltvfaZcRcqpQe3K3qNSxEEjV/0Hq1ylapozop+++P4X7T4cop1bNmrBnOmaOmWOWl9/rfJcJEOyf+cGvfnGa1odnK7br23pPXphwQISI4RntF0ODu1ar/dfeVbrwzy6uU1r79H/ReiRA/rl+2/Ud+Bs3XrvzRkryw35Hjekt6YsWK6gmdO0aW+SLm9ZxwhCrLp/85k27zuk8YNHKjV/BdWuVkZhB3fo6x4/aP+BYI0YOF7lazdUqYIp6t1/oHwKV1ClUqffOuxU/L/JGJUqX00VSxXUtEnj9f3v/bxHM7Bs3kz9+nN/xaSUUJ1qJxMCB/mLlleT2jUVHhxy9tGCb35d1eZyxR04piTqzFwgtH/wRf3y5ceU3zktatZvKv/cKYo1UetJQDj//EnLDmc9szNjZD8dCs+l5156Qx+89uyJpfiJYbvUb8BgRcWnq07DRib6ilFScsYKv/MPj341zjfcRIiXCsb8+bvWbdvsffe/qH/tIxr4208qlsX51lF7N+ijb4bo9nvvUOmi/zwfqXqT6zV8SE/VKl/QRPyhGjKgl3YmBajd1S10JDTMyPrZWf6lSxdoyOQ53ndnhl21V7KqOr/wglpUL64lm/bpqlvv19NPPSHPsXg1r1VRm1cuVuv29+uZh+9W4Lj+evS1T1WjcVvVzRdpyfeFy72ee9RrcpmRziSje/EqWKKyevYZqisb1/J++r/o9+PX2nfsmPfdmVGsfGVd3qCkjoeEe48Y5MqtJk2ayMcQJFabZhfhh7bqgUc6Kn+Fpur6ygt64KFH9ED7dtpjSGtAuUqqWtLHRPEx3m9noEKV2qpVqLBCw04+nl34586nlrWraufqxap/9S16ruPD2hI4QQ88/84pWQY2HK9o/Hai1m3d5T3yv6Difeqxndp36Lj3iLESiaF6992v1eyau9SiYT01qJxfK1etVKM2t+n+++7VAw88pGol8yk2o5zbRUG5SjVUt2gx057neSPzMyJV/YcP19q/aVsHZavWV4XSRW2plL9D3kLFVKVgmjas33CiL/esmaNf+k5Th6deVJdOd6lfjw80Y+1ezRj8q5buTNLzXV7Q9S0q6Z33XlVYfJx6ffuNkgvV1otdX1DVorF6+8Mv5AmopPYt6+r3H3/U3mNZa69/Z8YoLUkrVyzRmtVrtH1PsEqWq6h83vo7f4ejB3cpOS5SS5as1bV3PKjShfMoIcxEO8MGa11wuBo1vVZ3t2/FGI/5TpDWsXnntj0qW6Gi8ub218Gtq7RhZ6zuvv9m5U2L1bLlQQoKWmwiiUhVrlhJ/rl8tGXNMgUtW6ntO3eqYNHi2rx6mbbtPa6ypUtq86olWrc9WGXKVlBe32iNHzpdNz78uCoUSVXgvLlau26jdu09oArmesf379ScecsUHnpA8xYuU7Ey5VW4YH6lJsdr2eL5WrNll4oULaq1QYFW8QsElDQk+ojmzl+uYqVKKyHykD3n0lVr5RdQSgH+6Vq/ZqUORyWqWsVySoqL0JLARdq8Zat27z2o6OgIFShSRIEL5iqgiDGkuzdo065jqla1qlbMHqJX3vxSJao2Us0KJTKWzXsReXSfAucv1vr1GxUal67K5Usp4tgu/dTjdx2PzauGDaur3TVXq2ihfPKkJaj7Zx/pzzFBalC/tooWza85i5arZvniCgoMVFRiLrvliG96statWaKFi4O0edchVaxURXn8M9hcckKsVi4O1K6DEdq7eZVmzpmvXAWLK3jrWo2bOFWpfoVUqVxJmz5dYM65bs16OzRYpXJ5TRvxu975vJ9qNmygsMMHtGHjbpWvVN5EX5vMtdarVJmS2rZ2hTbt2KdNq5dr0859qla9itYa575smenH/cfNvVS25QNAfGyU1q9ba9o/WMf2bdDkmfPkH1BGhU1bL1k4X7sOHNeywFmKTPZV+RIBWrBgvmmn9dpzMMS0a2VtXzVHXV/7RCpc1ryvonyeWC0w8rR86RKFxEnlixXQurVrdOhYqMqUKqVNa5Zr1ZqtCj22T1OmzFSuoqVVtjilAf7Cb99+pLjijfXM/deZ59po5GGRUnMZIx4SrPkLg0wb+6hMsfxatmiejkUkGZ3YrcPHI1SqRElNGdxDn/8wRJXrNFPN0n4aPn6mGjSoq8Wzpyk60V9VK5UxV0jXGqMbq0y7bt68TcWNrKbHhmiekbUk3wIq6J+q+XNnKzrVX4khe/TGi88oPL2IGjWsryKFCpzIbyQnxWrhnHlabYzgEWPca9erq4YN6qtMyeIK37dRE5bs0zPPdlb92pXUrEV95feEaujYSerw3FtqbvpvrbmnhRvD9PrzD5vfVdd3732kCs2vU41yRe3504wOLw+ar9WbdqpwsaJav2Sh0ZmdyleohDwJx41uLVWREqWVHHNEC+bN0bKVa5WrQHHlU7wWzJmjo4YUzjD3l563qAorUjMWLNRq4xxT/QJUtqQTdaZr+9bNWr9ho44cD9PMqRN0KCxBFSuU054t6xW4cKXpq91avGqTKlWurG1rgrRo6Upt3bVXFSoj07l0YOcWLV62Qls3rdPCtdt1S/tb5Rd/xJDlfSpaooQK5c+nw/t2aFFQhj3yDyihXSun6+W3vlPJqrVVoVw5+RpHPmfRImMTlys2La8qlimhdPP8yxYFau2mbUae5utYfBE99tAt3vs2Tsg3WcNGjFaZarW13sjCvqPRqlq5nDauXqRFKzbKP28hFQ3Iq1XLF2nLgVBVrmD00psZ+PHd57T6aB793OtHE4372/IbpY2jLpzur/r1K5u2DlS4iik1JljL1m9XmUpVVaSgkeW5s3QsT1ndfUMrxYUf04KFi618H4tKUhWjs/t2bDZytNzYvH1asGSVylasbOyQaZ/lxu5u3ix/0z/FCxdQHk+kRo4dqxsffUmXNWmoPcbGTpy7XW+89rRsYXkv/PPk0dHgbdpwMEFP3PvXs2cGNmjV/NnKW7Gp2rSqZ4+N7fODgoKlN7o8boJIHyUd36rh05fpqa7vq1rpgsqbEquSxlfkjzP6vWSxDofEa+2SmQpatUXFSpbSohkTNW/xarvxdlETEB7YtVGLl6w09neN0vzyqWheP6vjm7ftVfDeLZoxO1ABZSvJJy5M8xYEKsU3wMrh/PnzFZ2Ux/iOIlYGFhsZWLp8hXLlL6yypUpqq+m3PSlF9MCtV+rI3o0KWm5s0drV8ilo5CaPr5aYc23ZutHYseOqW6uyUbp466/WG53bsn2f9TMJYYeNjVissIhwLZk/R2t37Dekq4oK5Dk5Pb1vx1rNMfZj3cYtKlqqgtbPGaYPPv1OJcwzVq1eUwWdlbKeNG1et0wrVq4ytnGLAkqWsXq/fskshfoU0x3Xt8343mlAyZN8icEaO3uHnuryiMj/zvqjp1Ye8dWbXR4w+lpe82cMV5RPUR0yMu1f5nLddXNL1a5eUn/0+1N+Bapo+cRxavvgw2paq6ry5ErUbwNG6/a771XDxo20YNRA7Y7Jq7befv47/CszRjNGDNSAYdMUnxirN194UoPGzfJ+8vegxlCHl15X3sTDGjGMzUylLRuWKTagnK5u01gJSRnj7eMH/qo+I6bLJ1e6RvX+Xm998ps9To0b0tdsebFg6mhNmr/CGLc0vfPCU+ozer52r5qtb38epFy+Kfrlm/f16Y/9tWLRVL304sc6GpaoHRsXq8uzb2nngUj5U0vGnJM6SKsmDtPLH/+m/P7J+uCtlzV3Y7CO7linD17rrO96/ampUydpT3DG+KxfLj8tnjlKj3XqqtDYRC2dMV7PPveGDkfEa9+WtRrYf5CJvo/qi69+VZKPv9bMGqdOnTrrcGySRvf5Tp/2+MWcxaOhfXqo/7i5SjZt+LG5zi9/jFBCWi7DCaONsi/Xth079EHXLpq6cLXdYiPCRJaR4WEnbb2SFnlM7733oTYHhyru8A7jBDtryuItSkyIU2xCglI8iQoJDz8xPybN45EnKUG+yVEKNcfTjPjFhAYrcMkarV++UK93fVU7DUHdsnSWBk6YrXwFcqvHhy/ro+8HZZzAwNc3XZMH99UzTz+rVRt3aP70MXr44Uc0LXClMU5z9Pobb+ugiUrH9v5F3ftNUu60MHV54XltOhSjXInRJkpOVVRUtLYsmavnn+isg1EJOmJIVdeOnbX14FEtmz1Fj3Z4ROMnT9L0qeM1cvQ4TTf9THT92SvP6I/R07x3Qv2jZE0a1FNPPvWkMcyLNOSXHnrkyU7acSRcg3/srk7PvKiZM6cqaOlC9fz2E01cuFG5DPnp8cHb6v7HJPkkx8k3LUUx0dGKDj+u0cMHaeXW/UqMPqRnHn9cUxas1soZQ/XOp18pKsVHQRP6q/Pzz2u+cVxzRg9Ul1e+OilLEmeIyJz5a9SiZSNLQOLC9uu9N57X8DkbFWsI7IevdNK4BWvlmxqt4YMGatWGXVq9eLqef/0LHTDyyT5vkYkpxjGF2bk94SEhWhhoyKn5znPPvaBQc7Glk4er+y8DzVd9NH1EP3Xu+oliYsLV74eP9bMh+kpP1ai+X+vr30coMt6jBJ/cio+JtBXBM6Pft59qatAOxQavVcfHHlTQ7kgVLpJB8tJsStbIih1m9FHhAOo+sW2NRxHmnjYsmaGFmzbrupuvt99X/vJqWiOvRoyZnPHeAL1aOne8Hnv6ReN447Vy7mR1fvZVBYfEKHjHRg3oO8DI5nF98eUvSvD4af38SXr66U6GcB/UT999pudMIDD1/9o7D+goq33t/9J77w2SACEBQmiBUEWKdBHpHURQsKJHFMvBjh5BegsBQpUeOtKli0gH6SmQhPReJ5nk/vc7oXm855z73fut7/OeedbKWpOZeefd5V+eZ+/97i3k8+DBA0Qvl+8W67h18QjDRkzgjjoJWEM1qUnXGDZkMF/Pi+HXE4eEfAwlZvMh7p49yBuTxrN0zUYh7wfZv3m5JNeT2FqbMnvaB3w+bzU5Kbf56tMvhZTkawkqLSFVO/7n7tWz/EV878rdZMqzE5g29Usypfl2rJ3Hy5Peo7SsFDNp5/z8fAoy7rNixTLupuWRnnCRUSPGcu52Kod3rGTBsjhDfDmXoIr6NOR/tW5mx579nD97jIkvDWfh2sPcvnSS0UOHc+ZKEqZm1axbJfHn8C8aQTBAx9FDl3APborf7wYk+wzpgaObCyamZZwXEnhViPO3n0xl2UbDiKHa8csw9VfN1x+9y7FrKVSXZfLBxEms2vULyVdOS73HSyxay4/790uc28Kn38yhpNpU+mc348e8xv3Cx+cKqH3QVHn3nT5Nxx5d/nBa65+Oh0k7VIu9PbpUn8+BI4cJahyF1SOWZUqxiKB1KxZJfI3hnckTuZFVIbGskG/fe5MPP53O/dQ05n3zMePfnMJ1EevrhVx9OXepkMNMPn7vY05dS+TuxUO8O+0bskv0/Lh6NhPfmszPl2+yNWY+701bKMFdxzcfTWFR7CEsRGDM/+ZTZizYSVFWivTxCtIKKrh74RgjR00gIT1XCKmZuIdK4VV8NuVt8U9zLh7dzRkhPick7s/dcAhbSz2LZ6zRarF5+RyJX/slhupZt/A7pn69TFu7M+fzyfxV2vn2nbt8OWUKa7cf077/EFl3LjJn6SpMrazZ+8MCRr/6kYhePRUmZuTnZlOsNgGsQXbKTSZPfJe72Xq2rJrFjBWGPPuvPuKl5Yia1woOIg6T02/xW3qRcmqNhJeXVWHpaCHx/4jWvyaWVtqecGqdm51jBcfPXdGuNbe0EIFQVbO3oBktmwaxe+e2f2mW5k9HjCpzH7Buwz7aPNebwYNHsiJ2NR2bhdZ8+o+h1snYuwbx7vg+bFi/TghKNnv2bqfLgFFYV5UJETKnIvcey5ZvoVvfF+ndqx8TJoxg+/IlJAjxUJsUqvnUspIilq2IJelernSSCY2D66EvTmfF/GhqNe9Jn76D+NusJYwb3JeoyFa4KwogndeyTTu8TdR6i5oCCVRwUsrv1VfG4ufmQLmQist3cmgeGUGApx+Dx7/L2pWxtImoqaOZpXz3NSJ8LbiXW02XXt2xLEwR4yyhuLKKnsNGwe1T7JQgrBZvetSqg7utGI2FB23btcHVyYaqkizOHDtISFQHXhj4Im0auuJVNwR/T1cxMDPadevHR+LsUVF1uZmYTGTXZ3Hz9KLfgIEE+j1e83Dypx2clLKOGzOMUW9PpV2QHd8vXIZPYARRoSGER3ail1z7MGCZm9sSGdmUenXC6NOnJ/ZCPM1tnRk4aiLTPv2rKHszHqSnsXb1cq7dyqS0SEd43TAq8jIfBThzK0datWlCncBgJv3lL0yZ/Dou9lWMmPAKH384BUeLPLKFwIVFtuGlkQNxtLWgMC+PGynldGjXDC/3APoP7s/znTviZu9MtakFjRuG4+/mjrm1M62fiSKsXiAffrWA2Z9/yBEpS7wkwsKyahqGh1IgyeghNXRy8ZB6huIV3ISPp01n25ZYdPducDVeR9vW9WnarC1L18YxsE19YtfuY+z4sfR5cRSD+7VjyYz5+DdqTEitAJ7r0Rt/FzOWrd5CTrokXVMHQr090JlY0aXzM0LEq3BwsaFj68Z4eIQxafJkvv14AqVJtyl+wsezhWzGF+gfrZFrGNWbkf26cvPKr0S0akGTsNokxieTnJJO3WYtGTKiL8916Yi7/L6ZjSMdu7YXBRjAoIE9cXe0wVKU6aDRE5kz52sh+yXcv5cmdr+MkM7KN3oz9e1xXD+8hpuF9jRr3FBbDGvr5EVk0wjtdb3Gzagnyq1Vh640qS+KWCsVlD64zLIthxj39uuMef0vLJr9PSHe/2SNmVysiNKxnZvZtu80r039hjnTXqn5UPrCz5c7vz0xrC+iYML4SbSoZUuiJLEuvXpiVfxACHsxxRWVdB86CsvEX9i+6xj5RaW4+QXh4WCJrWcwofUDadnlebZv/YGoWubEbjxMaU4ejk7++Ik9FagzBjWY0bZDGwLtXBk8ahJLVm9maJs67N/1Ey3bt8DDpZYkwhmsWjyDk3FbuHU3nbz8QsIaNxabzGLv1pUkmngyZtww+g8bQ4MGAVRKIG/etishgWKPZqYc3rCeVHNfRg94nm9nRfP+G+Np3aIR/l7+9Ov/IublmcRu+FGSZx5Wtp4EuziSnZHIgu+jad9zOL16duft0d00UvJ7qGQ1YuxEFi5fz6TBz/BD3DYGjXuTkX3bcPzCVXIfpODhVY+XRzx5yHAFZZJnrKyfXs/zGNVU6kxoGtmZqR9PY2z3JiTfeHw2oZmFJcnn9rDtyG3eeG0cA0a+RqeWfiyaG0tEmxb4uPnx8uRprIuN5trRH9FZ1WPw8734dNr7FGVeYu3Go5IfDccN/fzjDjbtOMTgCR+wbMZ7j8nNfwPlQrzTEpMICn4c51TbqTMtQ5s0p2WklNHHG4sqPR6BYUSE+dCkRQden/w+Lw3qKj5jwZQPPuCV0S8I6bhJpdhItxcHSLyJUo+Ac+9+OiXmznSKCsfXvwmT332HL94ZSvpvv+HmUZcmvr5YVFdi7ShkX2Kco1U1v50/xe64g+LfebhITnAW8pmbJyKvZgRP6AS/XU9mZ9xeeg99nd7PtEKXl8TJEwdIq/LibzPepTTjDjEr4uj14kB69X6R8S8PYXP0Ikw8/GgSEkLzqO58/NlnDHs2jPt3nl6ov1fE0MFTdykXQVk7oD6O1eWEtmhB7eAgnuvZh2Bvw8kPCmbW9hLPR9I2or6UyoQLIr7+O2j74nACyrN4/823mT7tTXb/egVHDw9Gjx1L/tUTEgs/YbIQzHupSTRs24r+/fuzb9FMPv1qOt/M+JpcsTdHO8NO/i7+ftwX0lqodgv+J/jTESM1EpGUlSps1ZAq27RrR53aPggn+KfQ1IqQnx4DXsUu6xKz/jaVGyUNebZxkHZmmdrtOCM7l6SUHKwsDEODoQFB+NhDofbUgkHvqKAvsZW+QoCGDB/N2p07eXNEJ25lZpNTXqxd16xlGxoEeVOtLxcipK5Sfxq/rfl7DGt7G04d2s4vCVl4eHuiUwxc9YyobW83RyytJEnVlEfBsXZTurSoT/TM77goQcfDJ4C5s77n9MmLtO/SXRw7hcCmLRk5fAgffvEtW9evJEBypdphW0VDU1sPmrVox/4dW/j59BmyzPxp2aQp5tUV2vEhzo6GnaHN7ezE+eQanU5LSurpoCeRev8OpdIfpjVMr0GTUCFjhnnkanFuvf7vn3hRu4GrpzxUUFYBx9LaToiNJaYWZljYCGmT+xQUldC9ez8GDR5G9IbNfDl13NPrukyrcHKSxCG3VbuOqykHG2kfFSgsrawMO5tbIkRlA2mVFtjZ20uwVvfVa4uv1bFzak9r9ZtPLiY3xJlKXFzcsLGxx0p+R4cl3V/orznimo1xvD78eSVrtO8rmKlDgx38cLI0wSs0kjB3Kwok6ZmJ2nZ398FSypiVmy9BrfTRFFxkSANsKCC/uEI7aFYdVlxcUoKtIomDhjN2wuts3buNPh2bUi6G9vBuyo6sRLU5iVC0sDSXdlfnuj2G6h/dI3szYOjAftw5sYcFW09iZ+fBpUPr2HXsDFZ2wbiYK2IuQV++p36nQj1tI+2jramUN6xtrHB1EcOxtBTRYEFxdiq52SlCcA2BJjg8HFdPN+4LmauUi7SzCAWqbx/GbENfP1lK8eGsRErL88XnxD7MnejxfF9sNVX3OzztJtKvZvQYNppPpk3jjQmjcLV/nJzNpHxVxU+PStn7h/NcVAOWiW+cu3oNTyE/C+fN4ejhX+jQrZfmJwERzRkxfCgffCbEduMaartaiX1U4yN+qFCpKycgoB7Dho3knY8+Y/vO1dT3c9M+0yBVsxcb9vD2l39Mhbw2EXfJo1xc3czMGi817SYEt6Cokg49BzB8xBiiV63jqzeHknrnquHQZ7nSRJK9uY2qT7Vmx0qkqUa8k3KPshoq7hsUSveOLbUDpFU/KbspKCoUshzAkKEjJUm8z/a9m2hW24Yr94uFyBkSlrWN+MbvG1Og+sjb0zD12KhFU3SVJVJoR8aNGsRPGxexdvePlFra0ijI8B0D7AjytiI36Sa/W4lISX629tRgVbUpzs419xbbeWI8RtrElOt3EykpFburWf3QrkE4VULwysQWTU2t8NamKk25k10g4sAwpWPuXo8wBxey0lO1KT1lU50HDBXy9VfeeXMC3i7/9QXkfwSdrlIbFX84baig7mUh9Wgc3pSw8Ca8++FfCfKwFn/TYS4O7uxssBUb8U0XNw/ttRrNsZAgZC6xu6LogUaaXQODqJJEVaXih/iKrY0tqsctrcylL2p8RxPOhnur+KjKoUar3Gs1YtTYcbz9wafErY2mQS3JEyoJaTnFgpkiLnJuHOaF/oPZvOdnOjw/lld6NuOdYT2ZOnMJlxPTSRaB9zCvhQUE42WnF8Gn1o1KDNX6y1TKoj5/2layc3KJFPI3dOgwvpizSEjrd5jr1SHrf++zFuKHd+JvEHf4JD5qWcQfuPV/BU6+YWw/cZxp702inq+X+Hwtej3TjtqRPTj281EmjByAraUV/vU680yoD30nfsDBQ3H06dwBK70FvTr1frTcQOUFSsqFKzwdj/4Iht74E0Gtq2nkY03M0uXcF8c5vH0TO/f9JOomlV/PXaDiH9S5SlcmxlyFX90IhosqXrRgOaNGD5RP1BEeYgxinH7evjSq58zG3ce1azKyH2DtH4qvq6N2OKqJuTk2trY4WpSzZMk84pMzOPHTfjbuOE3DOgHsWzWLq4lp3LlwmOg1u0Uh2YndlWiPGyfevk6puZi7JP9qU+kkdVSDKK/lc76lyCKQl/p1kgBRKkEjn/TMHHEucSI1b/cHGDJsOEd2b+BChhVLoz/n7JFtpFR7Uk9imE/dQK4c3yqkYj9ZafeJmT+b+/nVQvzULQ3RyNHDFzd7My5dv82ISVPo16k5uvJyzXgeBjIzdcCp/EmBtWSXnpwpSf/xArr6Yc2oSL7L6RuJ2v/JqRm0jmimvZbIjl6M8PeQn6KsopyCBw/ILZd2l/KoGqo7qtfqWA9fV1til87m0s0krlz8hWXLN2lB/CFMxZErRbVo+dZEBVOpmHxB9aE0rZbgZ38znTqNO9EnqhF6uV9ufiaZWTnynQqycopFuedibauXBK0nNTONzALD9IiJ0ATRvFrisRRbc7ErY96CBdyUwHJq3za27f8J/RPEQ1VIjRaqolQV3idLZ6XNZ6vgpxeiqRBWtz5erjo27D6j/Z+cfh/PsJa4WpmiryoTXy0nJ7tIVF4Ci6JjSJdAFLdpE8dOXcRa7EMlE9VrioCqpKleKxKvnvSyeKIojo5O+AiZKSgykHOFOm16E+QodYhexSvTFxDuU8XyVVvp0vs57XN12K/cQGt7E6lLlSTblKQMSit0hr5XEULZqgRgZ796BNYO4ce49dq12elpYp8ORIX5YSE+UZRfLIKhgtt3lVIWUiGdoSurpEJIX25evpAkFcTBVYK8idR59neLKSgoYPOq+Zy6kqx9pmClEkW1+FrNIb4KlTppT2lTU0sbrd6/R3lJPq4hj49KeYiBQ4Zx8oAIgHuwdOkXXDy+k8RKN0JdTTQ/uXF6B4vX7RU/SdH85F5BhSQ0s0fD7X4+3tw8v5/l67aRlZ3J0sXLiRfV/yTUUTzFBbna6/jkdFHSTdGOmzOpkPZVv2OKr48zMbO+5vxvScRfPU3Mhj34CNE599NhEtIKtanCwpw8EV2GNjITG1PJMjK8PleO7mDb8UukJ95i3uw55ItbmZiUkycCorykkoyEsyxZsVYIeA6rV6zit3sFeFmXcvzYT9pvZecVSRIr1frkEZQjigPpqwwGdOP8VaKaG3y3VY/+RPhVM2feajr1Hqy99yRefvcd4q/9xNL1h2vegaKcByz/YRNZhSWaiHtILNQRQer4HwXVbyr2RTWOkHunseOQYcrjXmYqAY1a4yriQgmTKk30mtIqsDZXzuwlXfFCfQYZ0ictoiIlTqktHGpsQS4x/Pofw1zuqSXEGuhK1BrTX8gtqBFt0s5qeYKEdQ0Ons74BHqTLP74EBYiaDTiUmN3tm61if/1AIdPXcVahFiF3vAwh4n09UNxoGKR0CkSb1xiYcxGxo15mdpiFGoH+5T0VE0wPPyuirMPBZqdg7nEJPFffSEPctI0SuwmAuu3s/uIXbOFvOw01q1bw293k8VPRFBqvpnPmRTYdfAkb43sxIZ1q7hw8gi93/qOoyK4087s4WYaRNR3lhj0MK+lYlurIb7O9qoJ5He0t7WyqGOVnkRQgB9745ay6/BZUpMTmDN7KcWiLsuLdNKehZKrHm/vcl38afepO7z22qtaJM0vypN4XyZ9oI66MjRy0vWrXL72eBTxSVioHCCFedxjEtfcvISMmxG7bieT3v2QVqHu2vtK6NiW3+HE5ft8/uWnNU/tmlI3tBHJN46SWODEJ++O1cSpQnlhPg51vXA0qeT6+V9JlHz1n+HPt/jawgYvXzfWzJ/FqrXrScjSM1TUXMLeFbw/dyXd+g8T9V7z3SeQeu8m3383g9OX7tG8RXtCpKFLnOozblAPzgnBiF6+lkQJTE3ad6RzZENWLprJnQeZnD51mr5jJxJZz415307n2KXbtGzdia5tw1ixaBZLYtdyNyWbF4eP47n2jdj9wyoWLFvJL+fiGfbyq4QEOLNr+2pWb4wjv6SMSxev4NcwhLsn9rJ2zxEJoo3wdqogekks8amZpCXcIi0zA2uzCuLidpCut+WZDh2wezIDCtz9A7h0Yje9XvqAzu0iOLdrG0PemUawlxMeQXXIvH2JGV9NJ+7ATwQ07kyvVrVY8v1Mjvwaz7Ndn+HKkT1s3bGf1JRkThz6kYMXk3EwK2R19ApxM2e8HEuJXbSA3CpR9J3a8eO6WI6cvUxU+w74uhumanxEdVTm32HOoh+ENCVyW/rioylvUZh6gRnzo4XYJFAnOJSQukpNG1CS94DVq+SzRAnTFalsXbsNS+866NIvszL2B+z9GjBueF92rVvM3OiVovRv0HfUeOoHGJRYRWkeyxfMYu/pazRs2ojz+zayadfP+IvqzLh+lJi1O3ELDsfVLJclS38gS5Ly7QtnqLB2oEfX9uzZupL9P9/luW7duHXlCEuWryEpIYGkDIkc4lgJV06zfu8JPOqG0bppY3z8PFg3dybLROVfTi7W1pPU9nqsoJPOnWDpDxsoqDRnr5D0uu0G8UKHABbO/J79l+NpFtWW0IbheNrA4gWztXUtp87f4PX336d+HW9+ObqLJVLm1s/2o31jd/42/WvWbdxKkSj3gX0lyC1ZwJ5Tl2kU3ogD+3Zx4OSv1AkJ4fjhA5JYjhES0VaIvKF9bZ3suXbqAFlW/nRp9fAQYSHy1Zlk48Ubo0RdFSWRbR3CuP7PymeVrJG6bTp0irAmUTSq68uKWTO4mfKAB4m32XfgBF7+wSReO8b6jTvxkIQ/4sWu7N4Qw7nryRw5coKW3YbzQueWFCXfY9m8uRw+e4H8jCzuCkGIbPcsZfHnWCT9WjeiFeEhhic+Ta1d8DQplCT/NzZs2UuxpReTXhmElQTPxLtXWTxrLsfOXcFFREq7lhGUFWayaN5Mdhw8Q2mlHU0ah+Di8OQIgY4F38yiY/+JtA5//Mi1gqt/LS6f2EX3sVPo0q4p5/fEMeitT6jr44JbYB1yEq4x44uviBPC69uwA1HBVsyeNZ/r8Sm0ad2F8KYNqMq9wyefTWf33h9xC2lGvx6dH43+UVXM3GnTuS1CpuDBNY4nlPLXqRM5un09K+N2YeYeROsWjbUF4ttXLGZh7BpOnL3BsHGT6BDZhJ9/XCMxZAtnr17i1tWrmNj7U5V1jehlm3ESEjpgcH9++/kQc2fN4eCJs7ToNITn2tUTW/uB9btO0a3PUBr5VjHtsy+J274LC49Ahg4eSqAnzBQbPHbiZ67cvMbN9HSaR7bTFjlrkOJfPHOSDXuPcP/WRZLzTZjyzmRcHcVQTa3xsczlco4Dn0wa/HfEo1ZoEzwdqpg/dxYnpZ9uX7vIfrGhJh274VmVyczZC0jILSXM15Xo2GXEp5VqNrpixUJuJebRZ/gI/CxKWRKzVJv2uxCfx0cfvc3lIzvFlzZTbO1Bh7ZRRESEcv5gHNuOXeLGz4dxqt+elwa2I3bx92zefUziqYX2Hfea0anfI/P+bWIWiC2dvUVYg8aEBPuTcfM4A8dOpWmrnpLwHTktcXBxzCpSCspo2aEz7i5u3JP6XEsopH/vjuSkJTF//gIOnDhP5oNkrl08y/pVi9hz7ILkkQg2xcYQX1hBw7pubIqJ4de7uYQ3rs+R7WvYdfw6zVq1JPHyadas30OFqZ6LJ47j6e/O8aNH+FXIQVj9euzcGceh05eI7NQTb7tCvp83jyu3bpEUnyTlKqVHvz446bP4avoMtu3eh29IS9qF12LO3BlcT8ihc5uGTP/mY8lDxUJWsglv1xmTlLN8MXcjzi6WZBbqee3tN2hYy43YhTNJSM8ScniGgeNfw9cym69nLSKzuIJ6nvYsWh5DYoaObr2642hrSKTB9epzS8jVF9/N4/DR40T2HETXpnXYt3U9W8Vv2nbtrS0DUSgvLSJ60UJOn7hIVdEDrtxJxFfqe2DzJi4m5PFcj45s+e5Llh+6yrAXu2nXPEROpoiTxdHsO34av9AIaZtg9AUZrF+5VnLGJtoOGMMb4wZpWi3l7g3Wr45l0/bTjH77fYlBLcROq7h+8VeWzpzHhaRs3v/iC5rWxByFrauWYevXSsRyCB9PHEuCuR8dm//xQuw/7c7XuZkPuJ+ahW+t2mLMjpTm5wjDLiIwsJbGgH8PpeoKCkSZSfMpZW0lRENNqah5/JLCQmHp5doCLlt7B+ysrcjJSOVBVgGuru54erpLZ+jJy82jUprLzs4BWxtLKUMa94U8efkG4OVuSJaFedkk3UvBxcNPVKKbFlSypKzpWXn4eHlTWlyMk5srVWqzsEq9KA5r7ZDT1JQUUd0OWJtWUSFq2U4UeGlpiQQpc5ydnZ9Y/PgYednpOAibViMIOaLeXTy9RNkYvqdGSVTCL8OSenWCMDPRUyCqXa+GpovT+XZGDB2eH0JUuB95oky+eH8G4z/5gEbBvtrcsLWo9jIhFSZmVjg5OVCcl0OhqH8vL6+nFEV1ZTmJCYmigyykHXxxsrMWRVdCgShHNRSspqTs7STY1qC6qpLMjAyqzW1xsBFyWlyGmbSBpVm1tsmauZUtzo722hoMdUq9s7s3/j6P5/vVSI46XFZNd9ja2Uo7llKmq8ZK7mFaVUFpWQWW1rbaPlOpD9Kxd3YV4V6KiaUtbi5O5AoxKSlXUyVeFOdLX6Wk4+HuJiU1PBUhmpZyMQxrG1ucHAyrS/Oy0rkn6sJT+tm7hhQ+xKHF3zJlRzw7l38q5a+gdu0ATKQcalM5tTeig4PYitiT1Jy05CRyRGV5eHjh7uqkqV31ZFtWbiHePr5aG6QJUc2Q/2vVDpJ2sJG+EZurqtamtdQUrl6UpppOU1NwanTGxlra1/5x+148sJnPlh1i8ZLZ2lOXCupwW7U9gpuzHfryIvLLTSUBqunSagpycynXV0k/2WlPG+ZkpgtdMtf6X220qA4xNqnWa8P2FmoaTwhJQW6W+F4Gjs5ueHt5aIq6ukJHYmICegvpPymPrqIKd9Wu0vZqetpdbNNGFO4jiB8p9ZlTqKN2UPCjPYjUJnpFmp9WY2ahpnUdNZspKMjX+lxNHzqKPapDfR/ixk/beW/eTlauicbVpoawPAHlJ/biJ+oK5SfOUpaHIxrq8FzlJ6VV5tQVPzGt1pFfUCR2Vo2Do4vWDlVi4/eSkiipqCYoMEjz14fQlabSvUkPxsxfy7N1bbBz9dbW8RVKHcrK1VSL+I/UQbmvajdl007uPgT4eBj6vzCXhIQUbBydsbY0wcbOSRSunuJSHVY2Nlp7V5QVEX83CXP5LDjQX4sphRLv8orKxI59MTepIPnePfKKdQQGBWEv/ahGZlLF3vLlPRcnO+2QYTdnJ6nP47KXl5WQIvZWJTHC29cbextrbYNN6XBWfTsNm8iRDOvWsObbT0Ot18yR+PdAkqy5pTXuYtMu4l/VFWXSfoVq+ETayZqSMvE98Sr1urSsWK4zxcFJ4q+5CfeTEikWX/Ty9pEy2lJcVESJxABTcwst5qkwUyr+oeKplb0jflJGC4nXBfl56CpVJDfVbOHJZQZPolJssrCwALUhr42tnRaHlC/cu/9ARF0gNpamlJYUU1wiZZTOsJc+UE8KZt08y5i3p/NdzDJCfRy00U5lDyr2qJEjNWVtYWWDg9h5odpc0NRMs4ly+Z1K+dxW2lHdR6cWA0s+qZT+y8wpFH9wlfoUYif11w41Fx+2lt/RiQ2q17Z2jvK/mfT1bcytHbExN8NU6qYOpVb5JykxkbJKU4KDg7AyrSa3IE9rT3uxkZLCfK2cFhL7fLy9qdYVkyY5R1degZvkHZeao4Sy01NIk7Joec3DXfPP3MIibfRKTQWWSM4x0XzMSXxMZRYDdKWFJCQmSax2EB+opY1oqhit7MvH10f6yuBPyi4y0lKRUCh5WWJFUbmWP8pLS9FLXnMSElsk8bS42lLzgSeh2rVAyqLXV2IpecBB4m+lEK37KZnaYfFubtIONXmwICeTnPwSnF3dcBZBqEHunSW5paikQuKNp/S3If4p5Cfd4M0pX/Hyh5/TPiKI1HtJWDq4ShkNhO73+NMSIyP+z5F0/Qwvj32JoNZ9adO0PmW52eSUWvPqq6Nwdf6fmav/d0ClrowFH7zB/L13OXh0C7Vr1mv8v4YayUwrdmP8+EG4O/1ni2T/dyA1/iqzYzbRa9hLPNPo6dGi//uoFoV6hB7dxjB5VixvDetU8/6fFXq2LvuOhRsP4ukVSXTMF9hbPiag/044sTOafdcLeWm0xMknRoiN+POhLD+N+UtX4VK3BS+90EkTFv8M/zY7XxvxGI4unjRs2Ehldm1tiY2TJ/0HvICH2x8PSRvxx6gQYlRl60jz5uH4B9TC+f+TA16bNG+FRVUJLp7e2P0vP4k/OyeLkMaRtGwQ+C8FvP9ZVFMoqrV+w4Y0CgnE29tLU9J/ZlSJWrd28mfMy6Pxc/9jNf3vgFohTfB2NMPUxhVXB8PDKEb8OVFaUoilgwc9OrV9ap+rfwTjiJERRhhhhBFGGGFEDf5+Qt4II4wwwggjjDDi3xRGYmSEEUYYYYQRRhhRAyMxMsIII4wwwggjjKiBkRgZYYQRRhhhhBFG1MBIjIwwwggjjDDCCCNqYCRGRhhhhBFGGGGEETUwEiMjjDDCCCOMMMIIDfAfoEUOyDpzL64AAAAASUVORK5CYII=\" width=\"582\" height=\"317\"\u003e\u003c/p\u003e\n\u003cp\u003eTable 2. List of transposable elements identified in the grass \u003cem\u003eRca\u003c/em\u003e genic region\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"640\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.125%\"\u003e\n \u003cp\u003e\u003cstrong\u003eGene\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.375%\"\u003e\n \u003cp\u003e\u003cstrong\u003eTE type\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.1875%\"\u003e\n \u003cp\u003e\u003cstrong\u003eSize (bp)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.5%\"\u003e\n \u003cp\u003e\u003cstrong\u003eRegion\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3125%\"\u003e\n \u003cp\u003e\u003cstrong\u003eE value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.5%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNo. of HSEs (sequence)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.125%\"\u003e\n \u003cp\u003e\u003cem\u003eObRca1\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.375%\"\u003e\n \u003cp\u003eMutator-like (MULE)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.1875%\"\u003e\n \u003cp\u003e336\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.5%\"\u003e\n \u003cp\u003ePromoter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3125%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.5%\"\u003e\n \u003cp\u003e2 (aaaaaattta; caaatttttt)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.125%\"\u003e\n \u003cp\u003e\u003cem\u003eOsRca2\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.375%\"\u003e\n \u003cp\u003eMutator-like (MULE)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.1875%\"\u003e\n \u003cp\u003e347\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.5%\"\u003e\n \u003cp\u003ePromoter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3125%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.5%\"\u003e\n \u003cp\u003e1\u0026nbsp;(aaaaaatttg)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.125%\"\u003e\n \u003cp\u003e\u003cem\u003eOsRca2\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.375%\"\u003e\n \u003cp\u003eMuDR-like (MITE)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.1875%\"\u003e\n \u003cp\u003e320\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.5%\"\u003e\n \u003cp\u003eIntron (5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3125%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.5%\"\u003e\n \u003cp\u003e1 (gtaatttttt)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.125%\"\u003e\n \u003cp\u003e\u003cem\u003eTaRca1-A\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.375%\"\u003e\n \u003cp\u003eGypsy (LTR)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.1875%\"\u003e\n \u003cp\u003e319\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.5%\"\u003e\n \u003cp\u003ePromoter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3125%\"\u003e\n \u003cp\u003e3e-43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.5%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.125%\"\u003e\n \u003cp\u003e\u003cem\u003eTaRca1-B\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.375%\"\u003e\n \u003cp\u003eMariner (MITE)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.1875%\"\u003e\n \u003cp\u003e239\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.5%\"\u003e\n \u003cp\u003ePromoter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3125%\"\u003e\n \u003cp\u003e1e-18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.5%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.125%\"\u003e\n \u003cp\u003e\u003cem\u003eTaRca1\u003c/em\u003e-D\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.375%\"\u003e\n \u003cp\u003eCopia (LTR)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.1875%\"\u003e\n \u003cp\u003e967\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.5%\"\u003e\n \u003cp\u003ePromoter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3125%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.5%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.125%\"\u003e\n \u003cp\u003e\u003cem\u003eTaRca2\u003c/em\u003e-D\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.375%\"\u003e\n \u003cp\u003eMariner (MITE)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.1875%\"\u003e\n \u003cp\u003e162\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.5%\"\u003e\n \u003cp\u003ePromoter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3125%\"\u003e\n \u003cp\u003e5e-21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.5%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.125%\"\u003e\n \u003cp\u003e\u003cem\u003eZmRca1\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.375%\"\u003e\n \u003cp\u003eClass II/III TE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.1875%\"\u003e\n \u003cp\u003e368\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.5%\"\u003e\n \u003cp\u003ePromoter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3125%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.5%\"\u003e\n \u003cp\u003e1 (agataattcg)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"13.125%\"\u003e\n \u003cp\u003e\u003cem\u003eSbRca1\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"24.375%\"\u003e\n \u003cp\u003eCACTA-consensus\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.1875%\"\u003e\n \u003cp\u003e584\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.5%\"\u003e\n \u003cp\u003ePromoter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"10.3125%\"\u003e\n \u003cp\u003e2e-97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"27.5%\"\u003e\n \u003cp\u003e1 (acaaaatttc)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"plant-molecular-biology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"plan","sideBox":"Learn more about [Plant Molecular Biology](https://www.springer.com/journal/11103)","snPcode":"11103","submissionUrl":"https://submission.nature.com/new-submission/11103/3","title":"Plant Molecular Biology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Rubisco Activase, Cereal Crops, Gene Duplication, Heat Stress, Photosynthesis, Molecular Evolution, and Regulation of Gene Expression","lastPublishedDoi":"10.21203/rs.3.rs-4676428/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4676428/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eHeat stress affects various components of photosynthetic machinery of which Rubisco activation inhibition due to heat sensitive Rubisco activase (RCA) is the most prominent. Detailed comparison of RCA coding genes identified a tandem duplication event in the grass family lineage where the duplicated genes showed very different evolutionary pattern. One of the two genes showed high level of sequence conservation whereas the second copy, although present only 1.5kb away, was highly variable among various plant species because of loss of introns, alternative splicing and loss of the last exon coding redox regulated C-terminal extension domain. Gene specific expression analysis, both at the transcription as well as the protein level, showed very different expression pattern of the two RCA copies. Expression of the highly conserved copy was higher under normal plant growing conditions that decreased many folds under heat stress with substantial genotypic variation, but the variable copy showed much higher expression under heat stress conditions across all grass species. The cultivated rice has only one functional gene as the second copy became nonfunctional due to multiple deletions but \u003cem\u003eOryza brachyantha\u003c/em\u003e and \u003cem\u003eOryza australiensis\u003c/em\u003estill have two functional \u003cem\u003eRca\u003c/em\u003e genes. Detailed analysis of the promoter region of the two copies among various plant species showed insertion of several transposable elements harboring heat responsive elements in the heat inducible copy of the gene. The conserved RCA copy of wheat didn’t have any transposable insertions whereas in that of maize has one heat shock element and sorghum had two. It would be interesting to study if the higher level of heat stress tolerance observed in sorghum and maize is associated with the differences observed for RCA.\u003c/p\u003e","manuscriptTitle":"Tandemly Duplicated Rubisco Activase Genes of Cereals Show Differential Evolution and Response to Heat Stress","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-02 04:26:41","doi":"10.21203/rs.3.rs-4676428/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Accept","date":"2024-09-28T17:03:31+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2024-07-10T05:43:29+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-07-09T14:21:15+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"Plant Molecular Biology","date":"2024-07-06T08:27:07+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-07-04T08:02:54+00:00","index":"","fulltext":""},{"type":"submitted","content":"Plant Molecular Biology","date":"2024-07-03T15:44:14+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"plant-molecular-biology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"plan","sideBox":"Learn more about [Plant Molecular Biology](https://www.springer.com/journal/11103)","snPcode":"11103","submissionUrl":"https://submission.nature.com/new-submission/11103/3","title":"Plant Molecular Biology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"45236d53-dfd1-4798-a585-9afb46314aa9","owner":[],"postedDate":"August 2nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-12-23T16:01:12+00:00","versionOfRecord":{"articleIdentity":"rs-4676428","link":"https://doi.org/10.1007/s11103-024-01515-z","journal":{"identity":"plant-molecular-biology","isVorOnly":false,"title":"Plant Molecular Biology"},"publishedOn":"2024-12-21 15:57:25","publishedOnDateReadable":"December 21st, 2024"},"versionCreatedAt":"2024-08-02 04:26:41","video":"","vorDoi":"10.1007/s11103-024-01515-z","vorDoiUrl":"https://doi.org/10.1007/s11103-024-01515-z","workflowStages":[]},"version":"v1","identity":"rs-4676428","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4676428","identity":"rs-4676428","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}