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Mosquitoes belonging to the Albitarsis Group (Anopheles: Nyssorhynchus) are of importance as malaria vectors across the Neotropics. The Group currently comprises six known species, and recent studies have indicated further hidden biodiversity within the Group. DNA barcoding has been proposed as a highly useful tool for species recognition, although its discriminatory utility has not been verified in closely related taxa across a wide geographic distribution.
DNA barcodes (658 bp of the mtDNA Cytochrome c Oxidase - COI) were generated for 565 An. albitarsis s.l. collected in Argentina, Brazil, Colombia, Paraguay, Trinidad and Venezuela over the past twenty years, including specimens from type series and type localities. Here we test the utility of currently advocated barcoding methodologies, including the Kimura-two-parameter distance model (K2P) and Neighbor-joining analysis (NJ), for determining species delineation within mosquitoes of the Neotropical Albitarsis Group of malaria vectors (Anopheles: Nyssorhynchus), and compare results with Bayesian analysis.
Species delineation through barcoding analysis and Bayesian phylogenetic analysis, fully concur. Analysis of 565 sequences (302 unique haplotypes) resolved nine NJ tree clusters, with less than 2% intra-node variation. Mean intra-specific variation (K2P) was 0.009 (range 0.002 - 0.014), whereas mean inter-specific divergence were several-fold higher at 0.041 (0.020 - 0.056), supporting the reported "barcoding gap". These results show full support for separate species status of the six known species in the Albitarsis Group (An. albitarsis s.s., An. albitarsis F, An. deaneorum, An. janconnae, An. marajoara and An. oryzalimnetes), and also support species level status for two previously detected lineages - An. albitarsis G & An. albitarsis I (designated herein). In addition, we highlight the presence of a unique mitochondrial lineage close to An. deaneorum and An. marajoara (An. albitarsis H) from Rondônia and Mato Grosso in southwestern Brazil. Further integrated studies are required to confirm the status of this lineage.
DNA barcoding provides a reliable means of identifying both known and undiscovered biodiversity within the closely related taxa of the Albitarsis Group. We advocate its usage in future studies to elucidate the vector competence and respective distributions of all eight species in the Albitarsis Group and the novel mitochondrial lineage (An. albitarsis H) recovered in this study.
Keywords:Albitarsis Group; barcoding; COI; new species; An. albitarsis G; An. albitarsis H; An. albitarsis I
The need to understand the systematics of the Neotropical Albitarsis Group (Anopheles subgenus Nyssorhynchus) [1] is primarily driven by the operational requirement to reliably distinguish which component taxa are involved in malaria transmission. The group is of great epidemiological importance as three of the five formally described species are proven regional malaria vectors in Brazil: An. deaneorum Rosa-Freitas [2-4], An. janconnae Wilkerson and Sallum (= An. albitarsis E) [5] and An. marajoara Galvão and Damasceno [6-8]. The vector status of An. oryzalimnetes Wilkerson and Motoki (= An. albitarsis B), An. albitarsis Lynch-Arribálzaga and the informally named species An. albitarsis "F" [9], are unknown. The group, along with An. braziliensis, is easily recognized in the adult stage by a pair of white scale stripes on sternum I [10]. The taxonomy of this group is historically complex and a comprehensive review is given in Motoki et al. [11].
Recent studies using COI and white gene sequences [12], in addition to the complete mitochondrion DNA [13] for five species belonging to Albitarsis Group, suggested that An. marajoara from Manaus (Brazil) should be regarded as a separate taxon in the Group (= An. albitarsis G) [12,13]. In the past year, two further lineages have been proposed suggesting that species discovery in the Albitarsis Group is far from complete. One, closely related to An. janconnae, was detected in the Caribbean region of Colombia based on COI, white gene and second internal transcribed spaces (ITS2) sequences [14]. The other, closely related to An. deaneorum, was found in Acrelândia, Acre, Brazil, using white and NADH dehydrogenase subunit 6 (ND6) gene sequences [15]. Neither of these proposed lineages was named.
Sequence data exist for mitochondrial COI [12,13,16,17], NADH dehydrogenase subunit 4 (ND4) [17], ND6 [17] and whole mitochondrial genomes [13], and the nuclear white [12,14], ITS2 [9,14,18] and Dominant receptor (D2) [18] regions. However, a single marker is still to be identified, which can to separate all known species and recognized lineages in the Albitarsis Group to ensure accurate species identification for studies on vector competence, for accurate distribution mapping, and to facilitate vector control efforts. Early attempts to use the ITS2 as a species diagnostic marker in the group [19] were hampered by the inadvertent inclusion of undiscovered taxa [9,13,14] and extensive intragenomic variation, which rendered the proposed ITS2-PCR assay unusable [20].
Mitochondrial genes are considered better markers than nuclear genes because of their abundance (1000's copies per cell), lack of introns, limited exposure to recombination, and haploid mode of inheritance [21]. DNA barcoding - which relies on the genetic variation within a standardized region of the COI gene - has been promoted as a reliable method for the identification of species in a variety of both invertebrate and vertebrate taxa [22]. Krzywinski et al. [13] sequenced whole mitochondrial genomes of six confirmed and putative taxa in the Albitarsis Group and showed that the 5' half of the COI "barcode region" is clearly more variable than its 3' half. COI barcoding studies of the mosquito fauna of Canada [23] and India [24], and within the genus Anopheles [24-27] have shown exceptional promise for species-level determinations. However, there has been no rigorous testing of the utility of the barcoding region in large populations of geographically widespread and closely related species such as those in the Albitarsis Group.
Herein we investigate the utility of the standard DNA COI barcode region for species identification using both the standard "simple" barcode methodology [22], NJ [28] and K2P model [29] to differentiate known and unknown species from 565 members of the Albitarsis Group, collected across a wide geographic range in South America in the last 20 years. For comparison, a subset of the full dataset is further investigated in a phylogenetic Bayesian framework [30].
Specimens and data access
Full specimen records (collection locality, coordinates, specimen identifiers, location of voucher specimens etc.) and all genetic data (edited chromatograms, consensus COI sequence files and corresponding GenBank numbers) are publicly accessible under the project code MBIK (Albitarsis Group Barcoding) on the BOLD website (http://www.boldsystems.org webcite), as part of the Mosquito Barcoding Initiative (MBI). Collection data and distribution maps are also publicly available through MosquitoMap (http://www.mosquitomap.org webcite). Specimens utilized in the molecular study were all morphologically verified as An. albitarsis s.l., using the original descriptions, and available keys [1,31] and include topotypic material for An. albitarsis s.s., An. deaneorum, An. marajoara and An. oryzalimnetes, and type series material of An. janconnae. Voucher specimens and/or their DNA extracts for the majority of specimens used in this study are stored at -80°C in the archive collections of Walter Reed Biosystematics Unit, Smithsonian Institution, Museum Support Center, Suitland, Maryland, USA, or in the Culicid DNA Collection of the Molecular Systematics Laboratory, Natural History Museum, London.
Sequence generation
DNA barcodes (658 bp, excluding primers) were generated for 565 specimens collected across South America using the published high-throughput DNA extraction and PCR amplification protocols [27] using the LCO1490 & HCO2198 primers of Folmer et al. [32]. Sequencing reactions were carried out in both directions with the Big Dye® Terminator Kit on an ABI 3730 automated sequencer (PE Applied Biosystems). Sequences were edited in Sequencher™ v4.8 (Genes Codes Corporation, Ann Arbor, MI), and translated to amino acids in MacClade v.4.06 [33]. Similarities with publicly available sequences were assessed using BLAST (Basic Local Alignment Search Tool), available at http://blast.ncbi.nlm.nih.gov/Blast.cgi webcite, and comparisons with unpublished barcode records checked through the IDS (Identification System) of the Barcode of Life database (BOLD, available at http://www.barcodinglife.org webcite).
Data analysis
To test the resolution of customarily advocated barcoding methodologies [22], 302 unique haplotypes (n = 565) COI barcodes were first imported into PAUP* v.4.0 [34]. Following construction of a pairwise distance matrix using K2P [29], a bootstrapped [35] NJ tree [28] was generated using 1,000 replicates to produce an unrooted consensus tree. Sequence divergence was calculated in MEGA v.4.0 [36] using the K2P model. Anopheles braziliensis Chagas [GenBank: DQ076238] and An. darlingi Root [GenBank: DQ076236] were used as outgroup taxa to the Albitarsis Group, following previously published studies [14,16].
A Bayesian phylogenetic analysis [30] was carried out using five specimens from each distinctive COI lineage found by NJ-K2P analysis (including respective type localities where possible). These sequences were added to the publicly available COI sequences of [14,16] (trimmed to 658 bp). Recently published COI data [12,13] overlapped those already included in our dataset and were therefore not included in the analysis. MrBayes [30] was performed online at: http://cbsuapps.tc.cornell.edu/mrbayes.aspx webcite. The Bayesian analysis was run for 10 million generations with two parallel searches using three heated and one cold Markov chain, with the first 5 million generations discarded as burn-in. The best-fit model HKY + I + G was selected by the AIC criterion in MrModeltest 2.3 [37]. The trees generated were edited in Figtree v1.2.1 [38].
Sequence statistics
MtDNA COI sequences (658 bp) were generated for 565 An. albitarsis s.l. collected across South America in the past twenty years: Argentina (n = 38), Brazil (n = 448), Colombia (n = 21), Paraguay (n = 23), Trinidad (n = 19) and Venezuela (n = 16). Among the 565 sequences were 302 unique haplotypes. Average nucleotide composition percentages for all 565 Albitarsis Group sequences were 38.5% (T), 15.8% (C), 29.4% (A) and 16.2% (G) (Table 1). Alignments were unambiguous: amino acid translations showed no stop codons, indicating that all sequences represented functional protein coding genes, not pseudogenes. The amino acid reading frame starts at the second base of the primer-edited sequences. The 302 unique nucleotide haplotypes, translated to 14 unique AA sequences (data not shown). All species in the Albitarsis Group were represented by the most common AA sequence, therefore AA derived phylogenies are not informative for species differentiation within this group.
Table 1. Mean pairwise nucleotide frequencies of COI barcode sequences for individuals belonging to Albitarsis Group.
Determination of barcode clusters
Following NJ-K2P analysis, nine distinct clusters were recovered (Figure 1), six of which corresponded to the previously determined species (An. albitarsis s.s. (n = 109), An. albitarsis F (n = 44), An. deaneorum (n = 36), An. marajoara (n = 70), An. janconnae (n = 96), and An. oryzalimnetes (n = 51), with a further three clusters corresponding to the previously described lineages An. albitarsis G (n = 105) [12,13] and sp. nr. An. janconnae [14] (herein called An. albitarsis I) (n = 12), and a newly detected lineage designated herein as An. albitarsis H (n = 42). Anopheles albitarsis s.s., An. janconnae, An. marajoara and An. oryzalimnetes were supported by high bootstrap values (BSV) (88%, 73%, 78% and 100%, respectively), as was An. albitarsis G (BSV 96%) and the additional lineage An. albitarsis I (BSV 92%). However, An. deaneorum, An. albitarsis F and the new lineage An. albitarsis H were poorly supported with BSV's of < 70%.
Figure 1. Bootstrapped NJ tree, based on 1000 replicates of K2P data matrices. Bootstrapped NJ tree of 302 unique COI haplotypes generated from 565 individuals belonging to Albitarsis Group, based on K2P distance algorithm. Bootstrap values less than 70% are not shown. *Highlights taxa regarded as species in this study. **Novel lineage. Outgroup taxa include An. darlingi [GenBank: DQ076236] and An. braziliensis [GenBank: DQ076238]. The bold lines indicate the nine phylogroups considered here.
Genetic divergence was estimated based on the nine clusters determined with NJ-K2P analysis (Figure 2, Figure 3). The overall mean distance (K2P) was 0.037. The average intra-specific genetic divergence was 0.009 (0.002 - 0.014) and the average inter-specific divergence was 0.04 (0.020 to 0.056). The most genetically divergent clusters were An. marajoara with An. janconnae (0.056) and An. janconnae with An. albitarsis G (0.054); while the most similar pairs were An. albitarsis H - An. marajoara (0.020) and An. albitarsis H - An. deaneorum (0.024) (Figure 2, Figure 3). Anopheles albitarsis I, with a mean genetic divergence of 0.045 (range 0.027 - 0.053), showed consistently higher genetic divergence with respect to all other taxa in the Albitarsis Group (Figure 2, Figure 3).
Figure 2. Mean inter- and intra- specific of K2P distances in nine species clusters. The species clusters were defined using the NJ-K2P distance method commonly utilized in barcoding. Numbers in boldface indicate intra-specific genetic divergence. Numbers underlined indicate the lowest (An. marajoara with An. albitarsis H) and highest (An. janconnae with An. marajoara) values between the species and lineages. Specimen numbers per species are indicated in parentheses.
Figure 3. Plot of K2P distance matrices of the nine clusters determined using NJ-K2P distances. Y-axis: genetic divergence and X-axis clusters. mar: An. marajoara; albG: An. albitarsis G; dea: An. deaneorum; alb: An. albitarsis s.s.; albH: An. albitarsis H; ory: An. oryzalimnetes; jan: An. janconnae; albF: An. albitarsis F; albI: An. albitarsis I.
Bayesian Analysis
Bayesian analysis supported all formally described species and previously indicated lineages detected with NJ-K2P (posterior probability P = 0.8 - 1), however, the newly recognized lineage, An. albitarsis H, is unresolved (P = 0.4, not shown) (Figure 4). The monophyly of the Albitarsis Group is confirmed and four species groups are recognized as follow: Group 1, An. janconnae, An. albitarsis F and An. albitarsis I; Group 2, An. albitarsis G, An. albitarsis H, An. deaneorum and An. marajoara; Group 3, An. albitarsis s.s.; Group 4, An. oryzalimnetes. Groups 2, 3 and 4 are more closely related, and they are paraphyletic with respect to Group 1. The lineage proposed by Gutierrez et al. [14] from the Colombian Caribbean coast showed strong correlation (P = 0.99) with An. albitarsis I. Lehr et al. [16] undetermined COI sequences grouped here with An. albitarsis G and An. albitarsis H (Figure 4) (see Figure 5), [16].
Figure 4. MrBayes tree generated using GenBank sequences and topotypic specimens and lineages found with NJ-K2P distances. Bold lines indicate the nine phylogroups considered here: albH: An. albitarsis H; mar: An. marajoara; dea: An. deaneorum; albG: An. albitarsis G; alb: An. albitarsis s.s.; ory: An. oryzalimnetes; albI: An. albitarsis I; albF: An. albitarsis F; jan: An. janconnae. Outgroups: An. darlingi and An. braziliensis. P values less than 70% are not shown. *Newly determined species. **Novel lineage. COI sequences from previously published works are indicated in blue [16] and red [14].
Figure 5. Map of South America showing the verified distributions of members of the Albitarsis Group based on COI DNA sequences. Locality data for 565 An. albitarsis s.l. specimens were plotted alongside those GenBank sequences whose identity has been molecularly verified within [14,16]. alb: An. albitarsis s.s.; albF: An. albitarsis F; albG: An. albitarsis G; albH: An. albitarsis H; albI: An. albitarsis I; dea: An. deaneorum; jan: An. janconnae; mar: An. marajoara; ory: An. oryzalimnetes. Localities determined from GenBank COI sequences are indicated as follow: An. albitarsis F [GenBank: DQ076234] [16], fuchsia diamond with heavy blue edging, and An. albitarsis I [GenBank: GQ153597-610] [14] black circles with bright green edging.
The analysis of COI sequences from members of the Albitarsis Group using NJ-K2P distances revealed nine distinct clusters: six of these correspond to the formerly defined species An. albitarsis s.s., An. deaneorum, An. janconnae, An. marajoara, An. oryzalimnetes and An. albitarsis F, and another to An. albitarsis G [13], which we believe to comprise a separate species in the Albitarsis Group along with "sp. nr. An. janconnae" [14], named An. albitarsis I. The remaining cluster, which we believe to represent a separate lineage, is named An. albitarsis H. Currently available synonyms for the Albitarsis Group as discussed in Motoki et al. [11] (An. albitarsis (Anopheles albitarsis var. limai Galvão and Lane, Anopheles albitarsis imperfectus Corrêa and Ramos)), An. marajoara (An. albitarsis domesticus Galvão and Damasceno) and the status An. allopha as a nomen dubium [1] are in our opinion sustained, with none referring to the newly recognized species/lineages presented here.
The monophyly of the Albitarsis Group is strongly supported with a high P value (P = 1), which echoes earlier studies [14-17]. BLAST searches of COI sequences revealed 99-100% homology with sequences reported for An. albitarsis s.s. [GenBank: DQ076204-DQ076208], An. deaneorum [GenBank: DQ076226, DQ076230], An. janconnae [GenBank: DQ076233], An. marajoara [GenBank: DQ076216] and An. oryzalimnetes [GenBank: DQ076210-DQ076215] [16]. This further reinforces our confidence in the identities of these taxa. The respective distributions and taxonomic positions of the informally designated taxa (An. albitarsis F, G, and I) and new lineage An. albitarsis H within the Albitaris Group are discussed in relation to earlier works below.
Anopheles albitarsis F
Anopheles albitarsis F was originally proposed as a putative new species in the Albitarsis Group based on ITS2 and white gene sequences [9] from specimens collected in Vichada, Colombia. One of the individuals assumed by Lehr et al. [16] to be An. albitarsis E (= An. janconnae), was unresolved in their Bayesian phylogeny (sample C14, Portuguesa, Venezuela [GenBank: DQ076234]), but this was confirmed as An. albitarsis F in our Bayesian analysis (Figure 4). Correlation of our data with these earlier studies confirms the wider distribution of the species outside of Colombia. As well as Vichada, Colombia [9] and Portuguesa, Venezuela [16], we further report it from the states of Cojedes and Zulia in Venezuela and in St George East and St Andrew/St David in Trinidad (Table 2, Figure 5).
Table 2. List of species found by country and state, belonging to Albitarsis Group.
Morphologically, An. albitarsis F is similar to the proven malaria vector An. marajoara [9], thus the wider distribution of this cryptic taxon in Venezuela and Trinidad is of epidemiological significance. Based on morphology and RAPD profiling [18], An. marajoara was believed to be the only member of the Albitarsis Group present in western Venezuela [8], where it is a proven secondary vector of Plasmodium vivax 210 [39]. However, these studies predate the discovery of An. albitarsis F and it is not known if the species-diagnostic RAPD assay (developed for the four known members of the Albitarsis Group at that time) [40] could distinguish An. marajoara from An. albitarsis F. Given the data presented herein, it seems likely that "An. marajoara" reported in Venezuela [8] corresponds to An. albitarsis F. It is also probable that "population C" from Venezuela and Colombia [41,42] identified on the basis of chromosomal analysis, may also be An. albitarsis F. Further sampling is needed to assess the true identity of An. albitarsis s.l. in Venezuela and assess whether An. albitarsis F is the only member of the group present there, or whether An. marajoara and An. albitarsis F are sympatric in that country. This is also true for the population of An. marajoara in Trinidad, which is regarded as a secondary vector in the region [43]. An ecological study stated that An. marajoara was identified from the island based on RAPD profiles and ITS2 sequences [43], yet our data clearly show all samples tested from Trinidad to be An. albitarsis F. Concurrent species determination, distribution mapping and vector incrimination studies are needed to assess the true impact of An. marajoara and An. albitarsis F on malaria transmission across their range.
Anopheles albitarsis G
Mean COI genetic intra-specific distance within An. albitarsis G was 0.010, whereas mean inter-specific divergence within the Albitarsis Group were four-fold higher (0.041) (Figure 2, Figure 3), and well above accepted species delimitation divergence thresholds routinely employed in DNA barcoding [22-27]. Based on consistent differences in DNA sequence in one individual (C10 [GenBank: DQ076225]) from four different DNA markers [40], Wilkerson et al. first suggested the presence of hidden genetic diversity within An. marajoara from Manaus, Brazil. Bayesian analysis of the entire COI gene showed that sample C10 grouped closest with An. deaneorum, but with low support (P = 0.69) [16]. Because of this, these authors questioned the validity of An. deaneorum arguing: "if An. deaneorum is a separate species from An. marajoara, then An. marajoara may consist of two or more species in Amazonian Brazil". The presence of An. albitarsis G as a cryptic species in the Albitarsis Group was later confirmed in further DNA studies [12,13]. BLAST of our An. albitarsis G sequences returned 100% homology with GenBank: DQ076225 (C10, from Manaus, Brazil, as An. marajoara) and GenBank: DQ076217 (C2, from Itaituba, Brazil, as An. marajoara) [16]. GenBank: DQ076221 (Itaituba, Brazil) [16] still shows deviation in the Bayesian phylogeny. However, at this time we are unable to assess whether this reflects further hidden genetic diversity, whether these differences are due to poor sequence quality in the first instance, or are indeed real and reflective of geographic distance as all other specimens sequenced are from Manaus, Brazil (Figure 4). Further sampling from Itaituba and other localities is needed.
Anopheles albitarsis H
This novel mitochondrial lineage was detected in 42 specimens collected in Rondônia and Mato Grosso, Brazil. COI sequences showed highest similarity with An. marajoara and An. deaneorum (0.020 and 0.024, respectively), both values being significantly higher than its intra-specific divergence (0.010) (Figure 2, Figure 3). BLAST of our sequences revealed 99% homology with GenBank: DQ076222 from Matupá and GenBank: DQ076223 from Peixoto de Azevedo (both Mato Grosso, Brazil) and GenBank: DQ076224 from Ariquemes, Rondônia, Brazil (as An. marajoara samples C7, C8 and C9) [16]. These three samples were shown to form a cluster distinct from An. marajoara, which is further supported in our Bayesian analysis (Figure 4).
Based on sequence variation, the barcoding community generally accept species delineation if the intraspecific variation is less than 1%, while the inter-specific variation is at least 2% different from its nearest congener [22-27,44]. Average inter-specific divergences between known taxa in the Albitarsis Group is 0.036, however, inter-specific values between An. deanorum, An. marajoara and Anopheles albitarsis H are much lower, ranging from 0.020 - 0.025. Our COI data supports the suggestion of Lehr et al. [16], exposing this grouping as cryptic complex in its own right within the Albitarsis Group. The status of the mitochondrial lineage An. albitarsis H is far from resolved. Further detailed ecological, genetic and morphological studies are necessary before we can speculate whether this lineage is reflective of a new species within the Albitarsis Group.
The presence of a cryptic species near to An. deaneorum (using ND6 and white gene sequences [15]) and near to An. marajoara (by allozymes and mtDNA RFLPs [45]), were detected in populations of An. albitarsis s.l. from the same and neighbouring localities to where An. albitarsis H was detected in this study.
Anopheles albitarsis I
Brochero et al. [9] first reported An. albitarsis F in Vichada, Colombia (east of the Cordillera Oriental) in sympatry with another taxon determined as An. marajoara based on morphological keys [10]. Microsatellite analysis of five populations of An. marajoara in Colombia revealed two incompletely isolated gene pools separated by the eastern Andean cordillera [46]. Using the entire COI, white gene and ITS2 sequences, Gutierrez et al. [14] reported a new lineage in the Albitarsis Group closely related to An. janconnae from northwestern Colombia. Sequences generated in our study share 99% homology with these COI sequences [GenBank: GQ153597-GQ153610] [14], and this cluster was highly supported in our Bayesian analysis (P = 0.99) (Figure 5). We clearly show that the lineage previously reported by Gutierrez et al. [14] as sp. nr. An. janconnae in the Caribbean region of Colombia and by Brochero et al. [46] as An. marajoara, is a separate species in the Albitarsis Group, which we call An. albitarsis I.
Despite the small sample size analysed, this species is clearly distinct from other members of the Albitarsis Group based on COI sequence data, with an intra-specific divergence of 0.008 and mean inter-specific divergence of 0.044 (0.027 with An. albitarsis F - 0.053 with An. marajoara) (Figure 2). That the sister taxon An. albitarsis F and An. albitarsis I were detected in sympatry in the geographically separate localities of Tibú, Norte de Santander, Colombia and Rio Socuavo, Zulia, Venezuela (Figure 5), provides further evidence to support their separate species status within the Albitarsis Group.
Distribution of Albitarsis Group in South America based on COI sequences
Locality data for all molecularly confirmed Neotropical Albitarsis Group specimens used in this study (n = 565) and correlated COI data from previous studies [14,16] were plotted using the mapping methodology developed by Foley [47] (Figure 5). This figure shows the distributions of An. albitarsis F and An. albitarsis I to be restricted to northern South America (Colombia, Venezuela and Trinidad), whereas An. albitarsis s.s. is found in southern Brazil, northern Argentina and Paraguay. Anopheles marajoara is present in the central east and west region of Brazil and An. deaneorum is in southwestern Brazil. Anopheles oryzalimnetes has a wide distribution in Brazil, mainly in the central region (Figure 5). Three species in the Albitarsis Group and the new lineage An. albitarsis H appear endemic only to Brazil to date: An. marajoara (Amapá, Mato Grosso, Pará, Rondônia), An. janconnae is present in northern Brazil (Roraima and Pará states), An. albitarsis G in Amazonian Brazil (Amazonas and Pará states) and An. albitarsis H in Rondônia and Mato Grosso, Brazil. Some species and lineages were found to be sympatric: for example, An. marajoara with An. oryzalimnetes and An. janconnae in Pará, Brazil; An. albitarsis H, An. deaneorum, An. marajaora and An. oryzalimnetes in Mato Grosso, Brazil; An. oryzalimnetes and An. albitarsis in São Paulo, Brazil, amongst others (Figure 5).
Whilst this study significantly contributes to the verified distribution of members of the Albitarsis Group, it is important to note that samples were not available from Bolivia, Costa Rica, French Guiana, Guatemala, Guyana, Panama, Peru, Suriname and Uruguay, where species of the Group have previously been reported [48]. We advocate the utility of the COI barcoding region for the correct species identification of An. albitarsis s.l. specimens in these regions, allowing complete species level distribution maps to be constructed for all eight species and the new lineage recognised in this study.
The results reported here using COI barcoding of specimens collected over 20 years from a wide geographic range in South America (Argentina, Brazil, Colombia, Paraguay, Trinidad and Venezuela), including topotypic specimens of the formally described taxa, helped resolve the taxonomic status and the distribution of the Albitarsis Group in South America. Contrary to intra-specific variation detected in previous studies using ITS2 [20], we consider the COI barcode region to be a robust marker of choice for species delimitation in the Albitarsis Group, not least because amplification of a short region (658 bp) yielded similar results to studies based on the entire mitochondria [13] or full COI gene [12,14,16]. We now consider the following species to comprise the Albitarsis Group: An. albitarsis s.s., An. deaneorum, An. janconnae, An. marajoara, An. oryzalimnetes, An. albitarsis F, An. albitarsis G and An. albitarsis I. In addition, we uncovered a new mitochondrial lineage, An. albitarsis H, which requires further sampling and sequencing of additional markers before its true species status can be resolved. This first comprehensive study of species in the Albitarsis Group across a wide geographic range in South America and provides a firm systematic basis for future studies that ideally should include ecology, biogeography, population genetics analysis, and vector incrimination of these species, particularly in malaria endemic regions where some of these new taxa may be acting as vectors.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
FRL, RCW & YML conceived the ideas; YML, RCW, MLQ & JEC obtained funding; FRL, SNM, JEC, MLQ & RCW undertook fieldwork and/or donated samples; FRL, YML & DML carried out the molecular laboratory work; FRL, YML, SNM & RCW carried out the data analysis and interpretation; FRL wrote the draft manuscript; YML, RCW, JEC, MMP, MLQ & SNM revised the draft manuscript; FRL, RCW & YML carried out the final revisions and submitted the manuscript. All authors read and approved the final version of the manuscript.
This study forms part of the larger objectives of the Mosquito Barcoding Initiative (MBI), led by the senior authors YML & RCW. This investigation received financial support from: supporters of the Natural History Museum, London to further the activities of the Mosquito Barcoding Initiative (to YML); the UNICEF/UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases (TDR) (grant A50252 to YML), Consortium for the Barcode of Life (CBOL) (to YML & RCW) and The Sloane Foundation (to YML & RCW); from the National Institute of Health (NIH), USA, (grant 2R01AI054139 to JEC) and COLCIENCIAS (grant 110134319196 to MLQ). We gratefully acknowledge Dr. Marta Moreno (Griffin Laboratory, Wadsworth Center, New York State Department of Health, Albany, NY, USA), Izis M. Carvalho and Galiza Primo (Instituto Evandro Chagas, Ananindeua, PA, Brazil) for their invaluable field assistance; Andrea Ormos for molecular laboratory support (Laboratories of Analytical Biology, Smithsonian Institution, Washington DC, USA), Dr. Desmond Foley (WRBU, Smithsonian Institution, Washington DC, USA) for elaboration of the map; Dr. Shazia Mahamdallie for helpful suggestions and discussions and Dr. Helena Brochero (Universidad Nacional de Colombia, Bogotá D.C., Colombia) for donation of An. albitarsis F samples.
This research was performed under a Memorandum of Understanding between the Walter Reed Army Institute of Research and the Smithsonian Institution, with institutional support provided by both organizations. The material to be published reflects the views of the authors and should not be construed to represent those of the Department of the Army or the Department of Defense. This manuscript was prepared while YML held a National Research Council Research Associateship Award at the Walter Reed Army Institute of Research.
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text:
Mother and Daughter Find Common Ground Atop National Podiums
Junior Brittany Sasser (left) and her mother, Connie, have each cleaned up on the national stage this year.
May 31, 2007
AMHERST, MA – Amherst College junior Brittany Sasser (Belmont, MA) returned home last weekend after exams to unwind for the summer. Brittany finished her third year at Amherst this week, and wrapped up her third season as a leader on the women's swim team. She helped the Lord Jeffs to a second place finish at the NCAA Championships in March by winning two individual National Championships and a third as part of a relay team.
In the Sasser household, the apple does not fall far from the tree. Last week, Connie Sasser, Brittany's mother, won a national title in the 200 backstroke at the United States Masters Swimming (USMS) Short Course Championships. The elder Sasser finished second in the 50 and 100 backstroke as well.
The USMS results are very similar to those which Brittany has recorded in NESCAC and NCAA Division III competition. Sasser '08, won the 2007 national titles for the 100 and 200 backstroke for the third consecutive year, and has never been beaten in the 50, 100, or 200 in her three years at Amherst.
Connie Sasser graduated from Florida State University in the 1960s, and was unable to compete in intercollegiate swimming, because there were few opportunities for women prior to 1972's Title IX. At the USMS Masters, she made up for lost time winning gold in the 50, 100, 200, 500, and 1650 freestyle, and in the 50, 100, and 200 backstrokes. She also took gold in the 200 yard IM, and silver in the 100 IM.
Brittany was the 2007 NESCAC Championships Swimmer of the Meet, and received 7 First-Team All Conference nods. She led the Lord Jeffs to a perfect 8-0 record at dual meets this winter, before being named the Swimmer of the Meet at the NCAA Championships.
After leading Amherst in points this season for the third straight year, Brittany was named the recipient of the Stefanie K. Duff '96 Award. This is an award given each year to the woman who made the greatest contribution to the team in terms of achievement, leadership and sportsmanship
Brittany is an Anthropology major, and is also active in an all-female a cappella group on campus. Next year, the Sassers will return to Amherst, once again, as Brittany tries to lead the Lord Jeffs back to the national podium with her mother rooting from the stands. Then, just after graduation, Brittany will return home to cheer on Connie in her efforts to repeat.

subset: commoncrawl


text:
Invited-Symposium Sessions
Session #
April 14
10:15am – 12:00pm
Grand Ballroom
April 14
1:30 – 3:30pm
Grand Ballroom A
April 15
10:15am – 12:00pm
Grand Ballroom
April 15
3:00 – 4:30pm
Grand Ballroom
April 16
10:15am – 12:00pm
Grand Ballroom
April 16
3:00 – 4:30pm
Grand Ballroom
Invited-Symposium Session 1
Sunday, April 14, 10:15 am – 12:00pm, Grand Ballroom
Order and disorder in the emotion brain: Emotional regulation in health and disease
Chair: Richard J Davidson1; 1UW-Madison
Speakers: Richard J Davidson, James Gross, Regina Lapate, Karina S. Blair.
This symposium will feature recent research on the neural bases of emotional reactivity and regulation and illustrate the application of insights gleaned from basic research to the clinical arena in understanding fundamental abnormalities in mood and anxiety disorders. In addition, new approaches to the treatment of mood and anxiety disorders, as well to the promotion of well-being in putatively psychologically "healthy" individuals, based upon knowledge of the neural bases of emotion regulation, will be considered. The following speakers will participate: Richard J. Davidson, University of Wisconsin-Madison, Organizer James Gross, Stanford University Regina Lapate,University of Wisconsin-Madison, Karina Blair, NIMH
Talk 1: Reactivity, regulation and recovery: The R's of emotion regulation from the perspective of affective neuroscience
Richard J Davidson1; 1University of Wisconsin-Madison
Parsing the temporal dynamics of emotional responding provides insights into understanding how reactivity, regulation and recovery in response to affectively salient stimuli can be disentangled. This talk will consider each of these subcomponents and provide illustrations from studies with both normal individuals and patients with mood and anxiety disorders/symptoms. Emphasis will be placed on prefrontal-amygdala and prefrontal-striatal connectivity, as well as on the temporal dynamics of amygdala activation in response to affective stimuli. Finally, behavioral strategies that alter brain function to improve emotion regulation for alleviating symptoms and promoting well-being will be described.
Talk 2: Emotion Regulation: Conceptual and Empirical Foundations
James Gross1; 1Stanford University
The past decade has seen an extraordinary increase in research on emotion regulation. Work from dozens of laboratories around the world has converged to establish that emotion regulation plays a crucial role in determining a wide range of affective, cognitive, and social outcomes. My talk will have three parts. In the first part of the talk, I will define emotion regulation and describe a framework for understanding the role of emotion regulation in healthy adaptation. In the second part of the talk, I will review key behavioral findings which suggest that specific forms of emotion regulation have different consequences for affective, cognitive, and social functioning. In the third part of the talk, I will discuss the neural bases of one particularly important form of explicit emotion regulation, namely cognitive reappraisal, and consider how adaptive forms of emotion regulation such as reappraisal fail in the context of psychopathology.
Talk 3: Investigating emotional regulation: Integrating psychophysiology and neuroimaging with an individual-differences approach
Regina Lapate1; 1University of Wisconsin-Madison
The remarkable inter-individual heterogeneity that characterizes emotional responding makes individual differences a powerful tool for the study of emotional regulation processes and associated neural mechanisms. In this light, this talk will underscore the value of adopting an individual-differences approach to examine the neural bases and information-processing conditions promoting successful emotional regulation. In the first part of the talk, I will report on neuroimaging and psychophysiological data suggestive of high stability and generalizability of success in a particularly effective form of emotional regulation: cognitive reappraisal. In the next part of the talk, I will examine the regulatory consequences of processing emotion in conditions precluding cognitive reappraisal due to lack of awareness of the source of emotion. Specifically, I will review recent findings demonstrating a benefit of conscious awareness in preventing physiological responses to emotional provocation from biasing appraisal of unrelated neutral stimuli, including the neural bases underlying those effects. Finally, I will discuss the relevance of this work for understanding emotional circuitry function and dysfunction with improved specificity under a biologically informed and process-based approach.
Talk 4: Emotional regulation in patients with anxiety disorders
Karina S Blair1; 1NIHM
Clinical reports suggest that emotion regulation difficulties are seen in a variety of anxiety conditions; e.g., Generalized Social Phobia (GSP), Generalized Anxiety Disorder (GAD) and Post Traumatic Stress Disorder (PTSD). However, the specific computational nature of this difficulty has only begun to emerge. This talk will focus on three studies of one form of emotional regulation; the role of dorsal prefrontal cortex (both medial and lateral regions) in attention control priming task relevant representations at the expense of emotional information. In the first study, patients with GSP, GAD and co-morbid GSP/GAD and healthy controls (HCs) performed an emotional reappraisal paradigm. Patients in all three groups showed reduced recruitment, relative to HCs, of dorsal anterior cingulate (dACC) and parietal cortices (PC) when explicitly regulating an emotional image. In the second study, patients with GSP, GAD and co-morbid GSP/GAD and healthy controls (HCs) performed the affective Stroop (AS) paradigm where task related priming of number representations implicitly reduces representation of emotional distracters. Again patients in all three groups showed reduced recruitment, relative to HCs, of dACC and PC as a function of task demands. In the third study, patients with PTSD, trauma controls (TCs) and HCs performed the AS. Patients with PTSD showed reduced recruitment of lateral frontal and PC when engaged by task demands while TCs showed indications of heightened recruitment of these regions. These data will be discussed in terms of emotional regulation deficits as a general risk factor for the development of a variety of anxiety conditions.
Invited-Symposium Session 2
Sunday, April 14, 1:30 – 3:00pm, Grand Ballroom A
Building blocks for language
Chair: Peter Hagoort1; 1Donders Centre for Cognitive Neuroimaging
Speakers: Simon Fisher, Ghislaine Dehaene-Lambertz, Kara Federmeier, Peter Hagoort
The language system is based on a language-ready brain whose development is instructed by genes for building such a brain, by a structural and functional organization of the newborn and infant brain that seems predisposed to develop a full-fledged language system, and by the recruitment of other than core areas in the perisylvian cortex to realize the communication potential of language. This symposium will focus on these building blocks for language in its full glory.
Talk 1: Decoding the genetics of speech and language
Simon Fisher1; 1MPI for Psycholinguistics; Nijmegen, the Netherlands
Researchers are beginning to uncover the neurogenetic pathways that contribute to our unparalleled capacity for spoken language. Initial clues come from identification of genetic risk factors implicated in developmental language disorders. The underlying genetic architecture is complex, involving a range of molecular mechanisms. For example, we have shown that rare protein-coding mutations of the FOXP2 transcription factor cause severe problems with sequencing of speech sounds, while common genetic risk variants of small effect size in genes like CNTNAP2, ATP2C2 and CMIP are associated with typical forms of language impairment. In my talk I will describe how investigations of genes like FOXP2, in humans, animals and cellular models, can unravel the complicated connections between genes and language. This depends on interdisciplinary research at multiple levels, from determining molecular interactions and functional roles in neural cell-biology all the way through to effects on brain structure and activity.
Talk 2: Neural basis of infants' language ability
Ghislaine Dehaene-Lambertz1; 1INSERM, Neurospin; Saclay, France
The development of non-invasive brain imaging techniques has opened the black-box of the infant brain and recent studies have revealed the early and complex organization of the peri-sylvian areas from the onset of the cerebral circuitry before term. I will discuss how these new data shed light on the emergence of language in the human species.
Talk 3: Many structures from the same blocks: language processing dynamics in the two cerebral hemispheres
Kara Federmeier1; 1University of Illinois at Urbana-Champaign & Beckmann Institute
Appreciating the meaning of a word, sentence, or larger discourse involves analyzing complex, often ambiguous perceptual inputs and linking them to information stored in long-term memory. Electrophysiological data suggest that meaning arises through a binding process, established via activity in temporal lobe areas and reflected in the N400 component, which creates an initial semantic representation built from activity in a distributed, multimodal set of brain areas. Although initiated by word perception, this process is not strictly feedforward. Top-down information built over time from context substantively changes processing by preactivating features of likely upcoming words. Strong temporal pressures are created by the fact that words are encountered at a rapid rate and that the binding process seems to be temporally constrained (N400 latency is strikingly invariant); information that is not available quickly enough cannot be incorporated immediately. Top-down processes that come on-line later thus serve to refine and revise the initial representation. The efficacy of top-down processing depends on the integrity of neural feedback connections, which appear to be different for the left and right cerebral hemispheres. In particular, left hemisphere dominance for language production attests to the presence of concept-to-form connections that are less well-developed in the right hemisphere. The consequence of this difference is a set of parallel, partially independent language comprehension systems. The multifaceted nature of meaning processing affords the brain important flexibility for dealing with varying processing circumstances, task demands, and resource availability.
Talk 4: The neurobiology of language beyond the information given
Peter Hagoort1; 1Donders Institute for Brain, Cognition and Behaviour & Max Planck Institute for Psycholinguistics
Speakers and listeners do more than exchanging propositional content. They try to get things done with their utterances. For speakers this requires planning of utterances with knowledge about the listener in mind, whereas listeners need to make inferences that go beyond simulating sensorimotor aspects of propositional content. For example, the statement "It is hot in here" will usually not be answered with a statement of the kind "Yes, indeed it is 32 degrees Celsius", but rather with the answer "I will open the window", since the listener infers the speaker's intention behind her statement. I will discuss a series of studies that identify the network of brain regions involved in audience design and inferring speaker meaning. Likewise for indirect replies that require conversational implicatures, as in A: "Did you like my talk?" to which B replies: "It is hard to give a good presentation." I will show that in these cases the core language network needs to be extended with brain systems providing the necessary inferential machinery.
Invited-Symposium Session 3
Monday, April 15, 10:15am – 12:00pm, Grand Ballroom
Memory on time
Chair: Howard Eichenbaum1; 1Boston University
Speakers: Marc Howard, Lila Davachi, Wendy Suzuki, Howard Eichenbaum.
The idea that memories are organized in time was initially proposed by Aristotle and was a central feature of Tulving's original characterization of episodic memory. There is now considerable evidence that the hippocampus plays a central role in the temporal organization of memory. Furthermore, recent studies are beginning to inform us about how memories are organized in time, and by what mechanisms the hippocampus plays its role in temporal organization. This symposium will review recent evidence from state-of-the-art research on this topic in human and animal studies.
Talk 1: What and when in the mind and brain
Marc Howard1; 1Boston University
The perception of time and memory are intimately linked. We consider the hypothesis that a variety of findings from cognitive and experimental psychology can be understood as operations on a gradually-changing representation of temporal history. Behavioral findings suggest three principles that this representation of temporal history should obey. First, at each moment this history contains information both about what stimuli were experienced and how far in the past they were experienced. Second, rather than falling off abruptly after some time scale, the history degrades gradually over hundreds or even thousands of seconds. Third, when an episodic memory is recovered, the brain "jumps back in time" to recover a previous state of the temporal history. We review neurophysiological evidence from a variety of species and brain regions and find evidence suggesting that each of these principles are respected in neural ensembles. These results suggest that it is possible to link cognition to neurophysiological findings across a variety of brain regions and domains of cognitive psychology.
Talk 2: Context and the human hippocampus
Lila Davachi1; 1New York University
Hippocampal function has long been associated with our ability to form, consolidate and retrieve episodic memories. Hippocampal processing is tightly linked with binding sequential representations to the context in which they were encountered allowing for recovery of the 'what' (i.e. episodic representations), 'when' (i.e. sequential information) and the 'where' (i.e. context). I will present a series of behavioral and fMRI experiments focused on understanding how we form mnemonic links across sequences of stimuli and how event boundaries disrupt sequential associations. Thus, representations spanning the same event become better integrated in memory (measured using explicit and implicit measures) and are associated with greater hippocampal neural similarity measures. Likewise, representations within the same event become reactivated during retrieval even when not deemed necessary for behavior. Thus, our perception of shared context modulates hippocampal neural similarity across sequential episodic representations and is related to how linked those representations become in memory. Further I will present evidence that hippocampal-prefrontal interactions support bridging across contexts to allow for higher-level temporal organization of our experiences.
Talk 3: Incremental timing in the monkey medial temporal lobe
Wendy Suzuki1; 1New York University
Recent work in my lab has focused on understanding how the brain encodes memory for temporal order, a critical component of episodic memory. To address this question, we recorded single unit activity and local field potentials from the medial temporal lobe including the hippocampus, the entorhinal cortex, the perirhinal cortex and visual area TE as monkeys performed a temporal order memory task. We found striking incremental timing signals from one item presentation to the next in the hippocampus. In contrast, the perirhinal cortex signaled the conjunction of items and their relative temporal order. These findings suggested that the perirhinal cortex might integrate timing information from the hippocampus with item information from visual area TE. We next asked if similar hippocampal incremental timing signals were also seen in other tasks that did not explicitly require memory for temporal order. When we examined hippocampal data from an object-place associative learning task we saw incremental timing signals very similar to the pattern seen in the temporal order memory task. In addition, we also identified incremental timing signals that also provided information about release type (early vs. late) as well as particular object-place combinations. These findings suggest that timing is a common signal conveyed by the hippocampus even in situations where memory for relative timing is not required.
Talk 4: Memory and time cells in the hippocampus
Howard Eichenbaum1; 1Boston University
Episodic memory is characterized by the temporal organization of events that constitute distinct experiences, and episodic memory depends on the hippocampus. Until recently, little has been known about how the hippocampus supports the temporal organization of memories. However, there are now descriptions of hippocampal 'time cells', principal neurons of the hippocampus that encode successive moments during an empty temporal gap between the key events. Time cells form qualitatively different representations ("re-time") when the main temporal parameter is altered. Distinct ensembles of time cells encode the key events and disambiguate different sequences of events to compose unique, temporally-organized representations of specific experiences. Time cells also encode spatial information and behavioral events to differing extents, but temporal signals are observed even when spatial and behavioral influences are eliminated. Finally, temporal information for hippocampal time cells may arise in the medial entrorhinal cortex, already known for its representations of spatial contexts. These findings suggest that the hippocampus segments temporally organized memories into events that occur at each moment within a temporally defined context much the same as they represent important events that occur in particular locations in spatially defined environmental contexts.
Invited-Symposium Session 4
Monday, April 15, 3:00 – 4:30pm, Grand Ballroom
Functional Specificity in the Visual System
Chair: Nancy Kanwisher1; 1 McGovern Institute, MIT
Speakers: James Haxby, Doris Tsao, David Pitcher, Josef Parvizi.
The visual system has been a primary battleground for one of the central debates in cognitive neuroscience: are some regions of the brain engaged selectively in a particular high-level mental process, or are all brain regions outside primary sensory and perceptual regions broadly engaged in many different mental processes? To address this question we will invite some of the major contributors to each of these views.
Talk 1: Distributed representation in the human visual system
James Haxby1; 1Dartmouth College
Visual cortical fields can be modeled as high-dimensional representational spaces. We model these spaces using a new algorithm, hyperalignment, that affords transformation of individual brains' voxel spaces into common, high-dimensional model spaces. Projecting individual data into common model spaces affords between-subject multivariate pattern (MVP) classification of fine distinctions among brain responses to faces, animals, and objects that equals or exceeds within-subject classification. Data in common model space coordinates can be projected back into the cortical topography of an individual's brain using the transpose of the transformation matrix for that subject. Building models with general validity across stimuli and across experiments requires broad sampling of visual stimuli, which we demonstrate using responses measured while subjects watch a full-length action movie. Models based on responses to still images from a moderate number of categories do not have general validity. Thus, category perception experiments do not provide adequate data for modeling the representational space in ventral temporal visual cortex. Category-selective regions are preserved in the model as single dimensions that reflect the relevant category contrast. The topographies associated with these dimensions agree well with the boundaries of individual-specific category-selective regions. The high-dimensional model, however, also shows that these regions have finer-scale topographies within them that afford fine distinctions among stimulus representations that are not accounted for by models based on category-selective regions. Thus, although category-selective regions are significant features of the high-dimensional model, they are only subspaces that provide an insufficient basis for models of visual stimulus representation in ventral temporal visual cortex.
Talk 2: Systems for Category-Selective processing in the macaque
Doris Tsao1; 1CalTech
fMRI studies in the mid and late 1990s described an area in the human brain that showed strongly increased blood flow in functional magnetic resonance imaging (fMRI) experiments when people viewed pictures of faces compared to pictures of objects (1). This seemed to offer an ideal potential preparation for tackling the problem of how the brain extracts global visual form: a small piece of brain specialized to encode a single visual form. Thus, 12 years ago, Winrich Freiwald and I began a journey into exploring the neural basis of face processing. We decided to look for a face-selective area in macaque monkeys, reasoning that it would not be unreasonable to find such a region in monkeys, since face recognition is also integral to macaques—and most importantly, if we did find such a region, then we could target an electrode to the region (something not possible in humans) and directly record from individual neurons to ask how they are encoding faces. In my talk, I will discuss the anatomical and functional organization of the macaque face processing system, as well as the more recently discovered macaque scene processing system. How are regions within these two systems system connected to each other and the rest of the brain? What representations are used in face and scene-selective regions? What is the contribution of different regions to behavior? What information is communicated between regions?
Talk 3: TMS evidence for category-selective cortical regions in human extrastriate cortex
David Pitcher1; 1Brain and Cognition, National Institute of Mental Health
Neuropsychological patients exhibiting category-selective visual agnosias have provided unique insights into the cognitive functions of the human brain but such case studies are exceptionally rare. To overcome the paucity of patients exhibiting category-selective deficits I have been using transcranial magnetic stimulation (TMS) to transiently disrupt face, object and human body perception in neurologically normal experimental subjects. Results support a modular account of cortical organization in which category-selective brain regions contribute solely to discrimination of their preferred category. Follow-up studies, that exploited the temporal precision of TMS, reveal the temporal dynamics underlying visual object perception in human occipitotemporal cortex.
Talk 4: Human Visual Numeral Area
Josef Parvizi1; 1Stanford University
Is there an area within the human visual system that has a preferential response to numerals as there are for faces, words, and scenes? We addressed this question using intracranial electrophysiological recordings and observed significantly higher response in the high-frequency broadband range to visually presented numerals, compared to orthographically similar (i.e., letters and false fonts) or semantically and phonologically similar stimuli (i.e., number-words and non-number words). This preferential response had anatomically consistent location in the inferior temporal gyrus (ITG) and anterior to the temporo-occipital incisure. This region lies within or close to the functional magnetic resonance imaging (fMRI) signal-dropout zone caused by the nearby petrous bone and venous sinuses – an observation that explains prior negative findings in the fMRI studies of preferential response to numerals. Since visual numerals are culturally dependent symbols that are only learned through education, our novel finding of anatomically localized preferential response to such symbols provides yet another example of acquired category specific responses in the human visual system.
Invited-Symposium Session 5
Tuesday, April 16, 10:15am – 12:00pm, Grand Ballroom
Networking Attention
Chair: Anna Nobre1; 1University of Oxford
Speakers: Michael I Posner, Sabine Kastner, Anna C Nobre, Earl K Miller.
Attention is a core aspect of our mental life, at the centre stage of cognitive neuroscience. This symposium brings together scientists working with different methods and at different levels of analysis to provide a contemporary view of the scope and properties of attention-related functions, and the mechanisms that make our cognition selective, flexible and adaptive.
Talk 1: Attention Networks Past and Future
Michael I Posner1; 1University of Oregon
Fifty years ago it was sufficient to show that attention changed certain operations in the information-processing stream between stimulus and response. Twenty years ago it became possible to implicate specific brain areas. Today papers examine putative brain networks that carry out some of the functions ascribed to attention and trace their activation and synchrony in real time. These networks are present in infancy but a long developmental process involving changes in connectivity is required to reach their adult state. Individual children and adults differ in the efficiency of attention networks in part due to different genetic polymorphisms that operate in interaction with environmental influences. The efficiency of attention networks can be improved by practice, and by changing the brain state in which they operate. Looking to the future, we should be able to foster the development of these networks, locate aspects that may be deficient in certain people and test methods designed to improve or eliminate the deficiencies.
Talk 2: Neural Mechanisms of Attention Control in the Primate Brain
Sabine Kastner1; 1Princeton University
Selective attention mechanisms route behaviorally relevant information through large-scale cortical networks. While there is evidence that populations of cortical neurons synchronize their activity to preferentially transmit information about attentional priorities, it is unclear how cortical synchrony across a network is accomplished. Based on its anatomical connectivity with the cortex, we hypothesized that the pulvinar, a thalamic nucleus, regulates cortical synchrony. To test this idea, we mapped pulvino-cortical networks within the visual system using diffusion tensor imaging and simultaneously recorded spikes and field potentials from these interconnected network sites in monkeys performing a visuo-spatial attention task. Here we show that the pulvinar synchronized activity between two interconnected cortical areas according to attentional allocation, suggesting not only a critical role for the thalamus in attentional selection, but more generally in regulating information transmission across visual cortex.
Talk 3: Attention at the Interface between Memory and Perception
Anna C Nobre1; 1University of Oxford
Top-down biases that prioritise the selection and integration of some events over others in the information-processing stream are a hallmark of selective attention. In most empirical work and theoretical models, biases are derived from the sensory environment and operate upon incoming sensory signals. However, arguably, the most prevalent source of predictive biases about relevant events to unfold is past experience stored as long-term memories. Building on the contextual-cueing and spatial orienting paradigms, we developed tasks to investigate whether and how long-term memories bias perception. Behavioural evidence confirms that memories significantly enhance perceptual discrimination and response speeds for target items occurring within remembered spatial contexts. Brain imaging suggests the involvement of the hippocampus, as well as the fronto-parietal network in mediating spatial contextual memory biases, and electrophysiological recordings reveal modulation of oscillatory activity regulating visual excitability in anticipation of target events. Overall, the findings cast long-term memories as proactive mental functions flexibly and dynamically adapting our interactions with the environment from early perceptual stages.
Talk 4: Attention is Rhythmic
Earl K Miller1; 1Massachusetts Institute of Technology
How are some thoughts favored over others? A wealth of data at the level of single neurons has yielded candidate brain areas and mechanisms for our best understood model: visual attention. Recent work has naturally evolved toward efforts at a more integrative, network, understanding. It suggests that focusing attention arises from interactions between widespread cortical and subcortical networks that may be regulated via their rhythmic synchronization. This could extend to all cognitive processes, suggesting our brain does not operate continuously, but rather discretely, with pulses of activity routing packets of information. Such discrete cycles would provide a backbone for coordinating computations (and their results) across disparate networks. However, it comes at a cost: it's naturally limited in bandwidth; only so many things can be computed or carried in a single oscillatory cycle. This can explain the most fundamental property of conscious thought, its limited capacity, which is the reason why we evolved attention in the first place.
Invited-Symposium Session 6
Tuesday, April 16, 3:00 – 4:30pm, Grand Ballroom
Emerging models of human and animal decision-making
Chair: Paul Glimcher1; 1NYU
Speakers: Michael Platt, Matthew Rushworth, Antonio Rangel, Elizabeth Phelps.
Over the course of the last 25 years cognitive neuroscientist have gone from knowing next to nothing about how humans and animals make simple decisions to having a fairly comprehensive understanding of what is coded and where when decisions are being made. From these insights a number of powerful models of how the brain makes decisions has emerged. We propose to survey what is known and to identify the frontiers that will be engaged in the next 25 years.
Talk 1: Neuronal Mechanisms of Decision Making in the Primate Brain
Michael Platt1; 1Duke University
Neuronal Mechanisms of Decision Making in the Primate Brain Abstract. A major challenge for neuroeconomics is to provide biological explanations of decision making that cohere across systems, circuit, cellular, and molecular levels of neural structure and function. This talk will review neurophysiological evidence for the computation of basic economic variables, such as value, utility, and risk, by single neurons in the primate brain. This evidence will be situated within a broader set of basic neurobiological mechanisms that appear to subserve a wide array of functions. The existence of similar mechanisms in a wide array of animals will be marshaled to endorse the idea that basic mechanisms are conserved and repurposed by evolution to generate adaptive behavior and cognition.
Talk 2: Neural mechanisms for foraging
Matthew Rushworth1; 1University of Oxford
Often when we design experiments to look at decision-making we ask our subjects to choose between limited numbers of options that are all presented simultaneously. When animals are foraging it is thought that they usually only encounter a single option at a time. The key decision is, therefore, whether or not this option is sufficiently valuable that it is worth engaging with or whether the environment is sufficiently rich that it would be better to continue searching for something better. The effort involved in making these different choices must also be considered. The anterior cingulate cortex carries three signals that are needed for such decisions – a representation of the value of any option that might be engaged with, a representation of the average value of alternatives in the current environment, and a representation of the cost of the actions. Considering the ecological context in which decisions arise also provides a new perspective on other signals in the anterior cingulate cortex, for example signals that are related to the encoding of risk.
Talk 3: The neuroeconomics of simple choice
Antonio Rangel1; 1CalTech
Neuroeconomics seeks to characterize the computational and neurobiological basis of different types of decisions. This talk will discuss a series of studies designed to understand how the brain makes simple choices, such as whether to choose and apple or an orange, as well as the quality of the resulting decision. This includes understanding how the brain assigns value to stimuli at the time of choice, how values are compared to make a choice, how they induce the motor movements necessary to implement the choices, and how these basic processes extend to more complex choice situations.
Talk 4: The Neuroeconomics of Emotion and Decisions
Elizabeth Phelps1; 1New York University
One popular theory of emotion and decision-making suggests that there are competing systems of emotion and reason that may drive choices. In contrast to this view, recent research in affective neuroscience has highlighted a modulatory role for emotion's influence on a range of cognitive functions, including perception, attention and memory. In this talk, I will outline how emotion's influence on decision-making may also best be captured as a modulation of the value computation. Specifically, I will present data suggesting that the emotional reaction to decision options or outcomes is linked to choice behavior, and how modifying emotional responses may change the choice. Finally, I will discuss the overlap in the neural systems of emotion and decision-making with circuits typically implicated in affective learning and emotion regulation.


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Unless you are an accountant or tax attorney, you probably don't know the first thing about a proper tax foundation for your startup. The very fact that there are accountants and tax attorneys suggests that the tax code is so dense and convoluted that we need a special class of people to devote their lives to understanding it just to help us navigate the system.
Your startup isn't about taxes; it is about selling the next big thing, or wearable widget, or consumer service that changes the way we… well, you get the idea. You're great at that. But if your startup is going to have any chance of success, you have got to lay a solid tax foundation.
Here are three items it must include: Continue reading


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MOUNT VERNON — Tornadoes can strike suddenly, with devastating impact. They can't be prevented, but you can be prepared if one strikes so that you have a better chance of survival.
Knox County EMA Director Mark Maxwell said the three most important things you can do are to make sure you have an emergency plan; stay informed, either with the media or Internet, but preferably with a weather radio tuned to NOAA; and put together an emergency kit, starting with a supply of everything you need to survive three days, including food, water and medicines.
According to the Center for Disease Control at emergency.cdc.gov/disasters/tornadoes, the safest place during a tornado is the interior part of a basement where, if possible, you should get under a heavy table or workbench. If you have no basement, an interior room is the best bet. If you are outdoors, lie down in a gully or ditch.
For the rest of the story
The rest of this article is available to Mount Vernon News subscribers. To continue reading, please log in or purchase a subscription. Click here for the May 23, 2013 e-edition. The article will only be available for thirty (30) days.
Contact Chuck MartinEmail
Copyright 2009 The Associated Press. All rights reserved. This material may not be published, broadcast, rewritten or redistributed.


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The City is encouraging community online members to keep an eye out and report graffiti whenever possible using the City's online request system, eService. Specific requests to report graffiti for removal can be made via the City's website or by using the Santa Clarita's eService iPhone or Android App for your smartphone.
Click the links to find out more information about the Graffiti Task Force or the Santa Clarita's Service. This app is available via iTunes or Google Play app store via the keyword search "Santa Clarita eService." Residents offering information that leads to the conviction of a tagger can receive up to $500 per conviction.
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Author: Katie Lloyd Thomas
Publisher: Routledge
Keywords: material, practice, matters, architecture
Number of Pages: 272
Published: 2007-05-15
List price: $165.00
Bringing together texts and work by theorists and practitioners who are making material central to their work, this book reflects the diverse areas of inquiry which are expanding current material discourse. Focusing on the cultural, political, economic, technological and intellectual forces which shape material practices in architecture, the contributors draw on disciplines ranging from philosophy, history and pedagogy to art practice and digital and low-tech fabrication. By paying critical attention to material, a wide range of issues emerge which are otherwise excluded from architectural d
Author: C. Tilley
Publisher: Routledge
Keywords: material, cultures, ambiguity, text, culture, art
Number of Pages: 192
Published: 1991-04-25
List price: $105.00
How are we to "write" an active interpretative account of past material culture? "Material Culture and Text" tackles this most basic problem of historical and archaeological analysis - the relationship between text and artefact - in an analysis of prehistoric art, fusing theory and practice with interpretation to create a fresh framework for understanding the relationship between past and present. It incorporates major tenets from post-structuralist thought in an analysis of prehistory. The text sets to work on a collection of rock carvings from Northern Sweden and shows how alternative concep
Defect and Material Mechanics: Proceedings of the International Symposium on Defect and Material Mechanics (ISDMM), held in Aussois, France, March 2529, 2007
Author: C. Dascalu
Publisher: Springer
Keywords: defect, mechanics, material, france, march, aussois, international, proceedings, symposium, isdmm
Number of Pages: 294
Published: 2008-04-14
List price: $149.00
This volume presents recent developments in the theory of defects and the mechanics of material forces. Most of the contributions were presented at the International Symposium on Defect and Material Forces (ISDMM2007), held in Aussois, France, March 25-29, 2007. The mechanics of material forces, originated in the works of Eshelby, provide a rational framework for the description of driving forces on evolving inhomogeneities and structural changes in continua. The general eshelbian mechanics formulation comes up with a unifying treatment of different phenomena like fracture and damage evolutio
Zinc Oxide - A Material for Micro- and Optoelectronic Applications: Proceedings of the NATO Advanced Research Workshop on Zinc Oxide as a Material for ... II: Mathematics, Physics and Chemistry)
Authors:Norbert H. Nickel, Evgenii Terukov,
Publisher: Springer
Keywords: zinc, material, oxide, workshop, mathematics, chemistry, research, physics, proceedings, micro, optoelectronic, applications, nato, advanced
Number of Pages: 256
Published: 2005-09-27
List price: $99.00
This volume reviews the latest experimental and theoretical studies in the field of ZnO. The chapters reflect the topics that were discussed during the NATO Advanced Research Workshop, which brought together researchers from Nato countries, Russia, Ukraine, and Kazakhstan to encourage cross fertilization and wider dissemination of the advances in this important research field. Fundamental issues such as growth of ZnO, structural, optical and electric properties, the role of the hydrogen and more were discussed.
Computerization and Networking of Material Databases (Computerization & Networking of Material Databases)
Authors:Satoshi Nishijima, Iwata, Newton,
Publisher: American Society for Testing & Materials
Keywords: databases, computerization, material, networking, amp
Number of Pages: 315
Published: 1998-02
List price: $99.00
IUTAM Symposium on Multi-Functional Material Structures and Systems: Proceedings of the the IUTAM Symposium on Multi-Functional Material Structures and ... December 10-12, 2008 (IUTAM Bookseries)
Authors:B. Dattaguru, Srinivasan Gopalakrishnan, V. K. Aatre
Publisher: Springer
Keywords: iutam, structures, functional, material, multi, symposium, bookseries, december, systems, proceedings
Number of Pages: 396
Published: 2010-06-09
List price: $159.00
This Symposium provided an international forum for exchange of ideas and creation of knowledge in recent advances on Multi-Functional Material Structures and Systems. Novel theories, mathematical models, analyses, and application of computational and experimental methods are the focus of the Symposium. In particular, it reflects the state of the art in mathematical modeling, computational methods, new experimental methods, new and advanced engineering applications in emerging technologies advanced sensors, structural health monitoring, MEMS, and advanced control systems.
Author: Michael Rea
Publisher: Rowman & Littlefield Publishers, Inc.
Keywords: constitution, material
Number of Pages: 408
Published: 1996-12
List price: $47.95
The only anthology available on material constitution, this book collects important recent work on well known puzzles in metaphysics and philosophy of mind. The extensive, clearly written introduction helps to make the essays accessible to a wide audience.


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Seven Hills, Ohio, August 13, 2012 – Chancellor University is pleased to announce that it has named Mr. Jason Wood CPA as the Theodore Ernst Chair of Accounting. Theodore Ernst, co-founder of the worldwide accounting firm Ernst & Young, was an early graduate of Chancellor University.
"Jason has done a superb job leading our undergraduate program in accounting," said Chancellor University President Robert Daugherty. "His dedication, expertise and enthusiasm in the world of accounting - as well as his commitment to higher education - make him especially well-suited for this role."
Mr. Wood is the Lead Faculty in Accounting at the rank of Assistant Professor at Chancellor University and has over 15 years of global, public accounting experience, primarily with the Big Four accounting firms. Mr. Wood also has over six years of academic teaching experience at various institutions where he has adjunct taught many accounting, accounting information system, and forensic accounting courses. He is known as being student-focused and strives to help students achieve their educational potential.
"Jason is a distinguished scholar who, by accepting this position, brings a wealth of knowledge and experience to Chancellor University," said Dr. Steve Kerr, Provost of Chancellor University. "We are delighted that he is the Theodore Ernst Chair of Accounting."
"I am honored and humbled by the opportunity to work to continue the tradition of excellence in the Chancellor community," said Mr. Wood.
Founded in 1848 as a mercantile college, Chancellor University is the second oldest business school in the United States. Notable alumni include John D. Rockefeller and Harvey Firestone.
Chancellor University is accredited by the Higher Learning Commission of the North Central Association.
Chancellor University is an independent, private, coeducational institution of higher education. The University offers academic programs in both business and non-business fields leading to Master's, Bachelor's, Associate's, and Certificate's in recognition of professional training and development. Chancellor University has a 160-year tradition of success in preparing students for careers in business and other professions.
Chancellor University provides the following documents in accordance with the IACBE (International Assembly for Collegiate Business Education) policies of disclosure on "current and easily-accessible student learning results for each of their IACBE-accredited business programs."
Business Administration,
Public Administration,
Paralegal Studies


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OKOTOKS, AB, April 24, 2013 /CNW/ - (TSX: MTL) Mullen Group Ltd. ("Mullen Group" and/or the "Corporation") is pleased to report its financial and operating results for the period ended March 31, 2013, with comparisons to the same period last year. For the three month period ended March 31, 2013, Mullen Group generated consolidated revenue of $385.5 million and operating income of $87.8 million. Mullen Group generated net cash from operating activities of $17.4 million that was used, together with cash from other sources, to pay dividends of $39.5 million, acquire net property, plant and equipment of $13.7 million and pay interest obligations of $3.7 million. Mullen Group's consolidated revenue of $385.5 million was a decrease of $40.5 million or 9.5 percent from the $426.0 million generated in 2012. The decrease in consolidated revenue was largely attributable to a $39.0 million decline in revenue experienced by the Oilfield Services segment, of which $21.8 million related to the non-recurring revenue generated by the design, build and commissioning of the Thin Fine Tailings ("TFT") barge system project for a large oil sands operator which was completed in the second quarter of 2012, along with a marginal $1.5 million decline in revenue recorded by the Trucking/Logistics segment. When factoring out the non-recurring TFT barge system project revenue, Mullen's core business revenue was down $18.7 million, or 4.6 percent. The Oilfield Services segment contributed revenue of $257.6 million, a decrease of $39.0 million, or 13.1 percent, from the $296.6 million generated in the prior year period. As previously noted, the majority of the decrease in revenue occurred in Canadian Dewatering L.P., which generated $21.8 million less revenue as a result of the completion of the non-recurring TFT barge system project. In addition, reduced demand for services by those Operating Entities involved in the transportation of fluids and well servicing, and the reduced demand for rig relocation services contributed to the decline in revenue. These decreases were partially offset by increased revenue recorded by those Operating Entities servicing the pipeline construction industry along with core drilling. The Trucking/Logistics segment contributed revenue of $129.1 million, which was a marginal decrease of $1.5 million over the prior year period. This decrease was mainly attributable to decreased demand for over-dimensional and heavy haul freight services. Mullen Group generated operating income for the period ended March 31, 2013, of $87.8 million, a decrease of $11.3 million or 11.4 percent over the $99.1 million generated in 2012. The decrease in operating income was mainly attributable to the Oilfield Services segment, particularly by those Operating Entities providing fluid hauling and well servicing along with those tied to drilling activity. In addition, the Trucking/Logistics segment recorded a $1.9 million decrease in operating income which was generally attributable to those Operating Entities providing over-dimensional and multi-modal transportation services. As a percentage of consolidated revenue, operating income decreased slightly to 22.7 percent as compared to 23.3 percent in 2012. "We knew coming into this quarter that it would be difficult to top the first quarter of 2012, which was a record in terms of revenue and operating income. Specifically, the completion of the TFT barge system project in the second quarter of 2012 along with the expectation of reduced drilling activity were both known going into the quarter. However, some extreme weather conditions in western Canada combined with bottlenecks in takeaway capacity with some of our customers in the heavy oil plays of Alberta were situations that we had little control over. As well, we witnessed some competitive pressures in businesses tied to the servicing of wells as a result of decreased drilling and completion activity in western Canada. On a positive note, a number of pipeline construction projects kicked off in the quarter which directly benefited Premay Pipeline Hauling L.P. and should for the foreseeable future, while Treo Drilling Services L.P. once again performed very well recording improvements both in terms of productivity and safety performance. All-in-all our results met our expectations," stated Mr. Stephen H. Lockwood, President and Co-Chief Executive Officer. In the first quarter of 2013, Mullen Group generated net income of $44.4 million, or $0.50 per share, a decrease of $14.4 million, or 24.5 percent compared to $58.8 million, or $0.73 per share in 2012. The $14.4 million decrease in net income was mainly attributable to the $11.3 million decrease in operating income and a $9.6 million negative variance in unrealized foreign exchange. These decreases were somewhat offset by a $3.5 million positive variance in the fair value of investments and a $3.1 million decrease in income tax expense. Adjusting Mullen Group's net income and earnings per share to eliminate the impact of unrealized foreign exchange and the change in the fair value of investments during the first quarter of 2013 results in adjusted net income of $45.4 million and adjusted earnings per share of $0.52, as compared to $54.0 million and $0.67 per share in 2012, respectively. These adjustments more clearly reflect earnings from an operating perspective. A summary of Mullen Group's results for the three month periods ended March 31,2013, and 2012, along with revenue and operating results by segment are as follows: SUMMARY (unaudited) Three month periods ended ($ millions, except per share amounts) March 31 2013 2012 Change $ $ % Revenue 385.5 426.0 (9.5) Operating income((1)) 87.8 99.1 (11.4) Unrealized foreign exchange loss (gain) 5.0 (4.6) (208.7) Decrease (increase) in fair value of (4.5) (1.0) 350.0 investments Net income 44.4 58.8 (24.5) Net Income - adjusted((2)) 45.4 54.0 (15.9) Earnings per share((3)) 0.50 0.73 (31.5) Earnings per share - adjusted((2)) 0.52 0.67 (22.4) Net cash from operating activities 17.4 54.3 (68.0) Net cash from operating activities per share( 0.20 0.67 (70.1) (3)) Cash dividends declared per Common Share 0.30 0.25 20.0 Notes: (1) Operating income is defined as net income before depreciation on property, plant and equipment, amortization on intangible assets, finance costs, unrealized foreign exchange gains and losses, other (income) expense and income tax expense. (2) Net income - adjusted and earnings per share - adjusted are calculated by adjusting net income and basic earnings per share by the amount of any unrealized foreign exchange gains and losses and by the change in fair value of investments. (3) Earnings per share and net cash from operating activities per share are calculated based on the basic weighted average number of Common Shares outstanding for the period. Operating income, net income - adjusted and earnings per share - adjusted are not recognized terms under Canadian GAAP and do not have standardized meanings prescribed by Canadian GAAP. Management believes these measures are useful supplemental measures. Investors should be cautioned that these indicators should not replace net income and earnings per share as indicators of performance. SEGMENTED RESULTS Three month periods ended March 31 (unaudited) ($ millions) 2013 2012 Change $ $ % Revenue Oilfield Services 257.6 296.6 (13.1) Trucking/Logistics 129.1 130.6 (1.1) Corporate 0.1 - - Intersegment eliminations Oilfield Services (0.5) (0.2) - Trucking/Logistics (0.8) (1.0) - Total 385.5 426.0 (9.5) Operating Income Oilfield Services 68.6 77.3 (11.3) Trucking/Logistics 20.1 22.0 (8.6) Corporate (0.9) (0.2) - Total 87.8 99.1 (11.4) This news release may contain forward-looking statements that are subject to risk factors associated with the oil and natural gas business and the overall economy. Mullen Group believes that the expectations reflected in this news release are reasonable, but results may be affected by a variety of variables. Mullen Group relies on litigation protection for "forward-looking" statements. Mullen Group is a company that owns a network of independently operated businesses. Today the Mullen Group is recognized as the largest provider of specialized transportation and related services to the oil and natural gas industry in western Canada and as one of the leading suppliers of trucking and logistics services in Canada - two sectors of the economy in which Mullen Group has strong business relationships and industry leadership. Mullen Group provides management and financial expertise, technology and systems support to its independent businesses. Mullen Group is a publicly traded corporation listed on the Toronto Stock Exchange under the symbol "MTL". Additional information is available on our website at www.mullen-group.com or on SEDAR at www.sedar.com. Mr. Murray K. Mullen - Chairman of the Board and Chief Executive Officer Mr. Stephen H. Lockwood - Co-Chief Executive Officer and President Mr. P. Stephen Clark - Chief Financial Officer 121A - 31 Southridge Drive Okotoks, Alberta, Canada T1S 2N3 Telephone: 403-995-5200 Fax: 403-995-5296 SOURCE: Mullen Group Ltd. To view this news release in HTML formatting, please use the following URL: http://www.newswire.ca/en/releases/archive/April2013/24/c8452.html CO: Mullen Group Ltd. ST: Alberta NI: TRN OIL ERN -0- Apr/24/2013 21:30 GMT
Mullen Group Ltd. Reports Record First Quarter Financial Results
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