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Characterization of ZBTB18 short band by MS.

GSEA analysis in BTSC233.

Proliferation, migration and invasion assays in SNB19 cells.

ZBTB18 promoter methylation analysis.

Glioblastoma (GBM) comprises distinct subtypes characterized by their molecular profile. Mesenchymal identity in GBM has been associated with a comparatively unfavorable prognosis, primarily due to inherent resistance of these tumors to current therapies. The identification of molecular determinants of mesenchymal transformation could potentially allow for the discovery of new therapeutic targets. Zinc Finger and BTB Domain Containing 18 (ZBTB18/ZNF238/RP58) is a zinc finger transcriptional repressor with a crucial role in brain development and neuronal differentiation. Here, ZBTB18 is primarily silenced in the mesenchymal subtype of GBM through aberrant promoter methylation. Loss of ZBTB18 contributes to the aggressive phenotype of glioblastoma through regulation of poor prognosis–associated signatures. Restitution of ZBTB18 expression reverses the phenotype and impairs tumor-forming ability. These results indicate that ZBTB18 functions as a tumor suppressor in GBM through the regulation of genes associated with phenotypically aggressive properties.Implications: This study characterizes the role of the putative tumor suppressor ZBTB18 and its regulation by promoter hypermethylation, which appears to be a common mechanism to silence ZBTB18 in the mesenchymal subtype of GBM and provides a new mechanistic opportunity to specifically target this tumor subclass. Mol Cancer Res; 15(8); 998–1011. ©2017 AACR.

Proliferation, migration and invasion assays in BTSC233 and JX6 cells.

GSEA analysis in JX6.

ZBTB18 promoter methylation analysis.

1. Climate change is causing shifts in the distribution of many species and populations inhabiting mountain tops are particularly vulnerable to these threats because they are constrained in altitudinal shifts. Apennines are a relatively narrow and low mountain chain located in Southern Europe, which hosts many isolated populations of mountain species. The butterfly Erebia pandrose was recorded for the last time in the Apennines in 1977, on the top of a single massif (Monti della Laga). 2. We confirmed the presence of a small, isolated population of E. pandrose in the Apennines, at a distance of more than 400¿km to any other known populations. Then, we examined the cytochrome c oxidase subunit 1 mitochondrial DNA marker of this species across the Palaearctic area and estimated the potential decline over the Alps and the Apennines due to future climatic changes. 3. The Apennine population represents an endemic lineage characterised by eight mutations over the 658¿bp analysed (1.2%). In the Alps and Apennines, this species has shifted uphill more than 3 m per year since the end of the 19th century and more than 22 m per year since 1995. Species distribution models suggested that these mountain populations will experience a generalised loss of climatic suitability, which, according to our projections, could lead to the extinction of the Apennine population in a few decades. 4. Erebia pandrose has the potential to become a flagship species for advertising the risk of losing unique fractions of genetic diversity for mountain species., funded by the project: ‘Ricerca e conservazione sui lepidotteri diurni di sei Parchi Nazionali dell'Appennino centro-settentrionale’. Support for this research was also provided by ‘La Caixa’ Foundation (ID 100010434) to M.M. (grant LCF/BQ/DR20/11790020), by the Academy of Finland to V.D. (Academy Research Fellow, decision no. 328895) and by project PID2019-107078GB-I00/AEI/10.13039/501100011033 to R.V., 1 Introduction 2 Materials and methods 2.1 Data collection 2.2 COI analyses 2.3 Species distribution modelling 2.4 Elevational shifts of E. pandrose records through time 3 Results 3.1 COI structure 3.2 Climate change effects 4 Discussion 4.1 The population of Monte Gorzano 4.2 What is the future for E. pandrose on the Apennines? 4.3 Erebia pandrose in the context of preserving Apennine butterflies 5 Conclusions Acknowledgements Conflict of interest

There are 96 endemic species of Eumolpinae (Coleoptera: Chrysomelidae) described from New Caledonia, but some estimates propose that the actual number could be at least twice this figure. Not surprisingly, when a particular species assemblage has been revised, the number of species in that group increases significantly. Here, we revise the New Caledonian endemic genus Taophila Heller, 1916, the best studied in this fauna and currently known to include eleven species, one in the subgenus Jolivetiana Gómez-Zurita & Cardoso, 2014, and ten in the nominal subgenus. The analysis of morphological differences in a large sample of Taophila and the validation of the resulting species hypotheses in an integrative fashion based on a phylogenetic analysis of partial mtDNA sequences (cox1 and rrnS) resulted in the addition of eleven more taxa. Taxonomic splits mainly reinterpreted the previous observation of mtDNA paraphyly affecting T. subsericea Heller, 1916, shown to represent a complex of species mostly distinguishable by diagnostic differences among females. The new species described are: T. bituberculata n. sp., T. carinata n. sp., T. dapportoi n. sp., T. davincii n. sp., T. draco n. sp., T. goa n. sp., T. hackae n. sp., T. samuelsoni n. sp., T. sideralis n. sp., T. taaluny n. sp. and T. wanati n. sp. These additions and the synonymy T. subsericea Heller = Stethotes mandjeliae Jolivet, Verma & Mille, 2010 n. syn., bring to 21 the total number of species in Taophila. Moreover, we also found the first evidence of mtDNA introgression between species of New Caledonian Eumolpinae, resulting from putative recent hybridization of T. subsericea and T. dapportoi where these species coexist. We describe a model incorporating the mtDNA genealogy of T. subsericea about the conditions that may have favored the secondary geographic encounter required for the hybridization of these species.

Aim: Occurrence data are fundamental to macroecology, but accuracy is often compromised when multiple units are lumped together (e.g., in recently separated cryptic species or in citizen science records). Using amalgamated data leads to inaccuracy in species mapping, to biased beta-diversity assessments and to potentially erroneously predicted responses to climate change. We provide a set of R functions (biodecrypt) to objectively attribute unidentified occurrences to the most probable taxon based on a subset of identified records. Innovation: Biodecrypt assumes that unidentified occurrences can only be attributed at certain distances from areas of sympatry. The function draws concave hulls based on the subset of identified records; subsequently, based on hull geometry, it attributes (or not) unidientified records to a given taxon. Concavity can be imposed with an alpha value and sea or land areas can be excluded. A cross-validation function tests attribution reliability and another function optimises the parameters (alpha, buffer, distance ratio between hulls). We applied the procedure to 16 European butterfly complexes recently separated into 33 cryptic species for which most records were amalgamated. We compared niche similarity and divergence between cryptic taxa, and re-calculated and contributed updated climatic niche characteristics of the butterflies in Europe (CLIMBER). Main conclusions: Biodecrypt showed a cross-validated correct attribution of known records always ≥ 98% and attributed more than 80% of unidientified records to the most likely taxon in parapatric species. The functions determined where records can be assigned even for largely sympatric species, and highlighted areas where further sampling is required. All the cryptic taxa showed significantly diverging climatic niches, reflected in different values of mean temperature and precipitation compared to the values originally provided in the CLIMBER database. The substantial fraction of cryptic taxa existing across different taxonomic groups and their divergence in climatic niches highlight the importance of using reliably assigned occurrence data in macroecology., Support for this research was provided by the Academy of Finland to V. Dincă (decision no. 328895), and by the Spanish Ministry of Economy and Competitiveness (MINECO) and Agencia Estatal de Investigación and Fondo Europeo de Desarrollo Regional (AEI/ FEDER), UE (Refs. PID2019‐107078GB‐I00 and CGL2016‐76322‐P) to R. Vila.

New Caledonia is an important biodiversity hotspot, where numerous plant and animal groups show high levels of species diversity and endemicity, while facing multiple threats to their habitats. Leaf beetles in the subfamily Eumolpinae illustrate this pattern, with species estimates higher than the number of described taxa and distribution ranges that are often consistent with microendemicity. In this study, we increase the knowledge of this group by focusing on the Taophila subgenus Lapita, known from three species but here expanded to eleven with eight new species: T. atlantis sp. nov., T. hermes sp. nov., T. kronos sp. nov., T. oceanica sp. nov., T. olympica sp. nov., T. ouranos sp. nov., T. riberai sp. nov., and T. tridentata sp. nov. Additionally, we infer the evolutionary history of the group using mtDNA markers (COI and rrnS). This phylogeny and the species distribution help hypothesize a model of evolution for this lineage in the context of historical climatic and geological changes of New Caledonia., This work was funded by the Spanish ‘Agencia Estatal de Investigación’ through Grant No. CGL2017-83324-P and the associated predoctoral scholarship PRE2018- 083172 to LP, both with support from the European Regional Development Fund of the European Commission.

The abandonment of agricultural land and the afforestation of grassland habitats represent major threats for butterflies in European and Mediterranean areas. A crucial goal for Lepidoptera conservation is to maintain and/or restore habitat quality by targeted management. Nevertheless, there are few experimental studies allowing to derive data-driven strategies to protect butterflies of open grasslands in the Mediterranean region. We developed a habitat management strategy for the conservation of the Italian endemic butterfly Zerynthia cassandra by adopting a three-step procedure: (i) characterising which environmental and host plant features influence oviposition on plants; (ii) identifying and testing the effect of a data-driven habitat management intervention; (iii) understanding which micro-habitat features promote Z. cassandra oviposition in restored places to optimise the intervention. Both patch (areas of 1 m radius hosting Aristolochia shoots) and plant features affect oviposition, with the strongest positive effects showed by high irradiation of the patch and plant quality (high number of flowers and leaves). Accordingly, the management consisted in vegetation cuts to increase irradiation, and 2 years of monitoring demonstrated that this procedure significantly increased oviposition (average increase of about 2 eggs per plant) and larval presence. Micro-habitat sampling demonstrated that the maximum oviposition differed between vegetation structures, highlighting the importance of a local fine-tuning before the intervention. We provided a data-driven, effective, and sustainable management strategy to increase habitat suitability and oviposition for an endemic and endangered Mediterranean butterfly. Our framework can drive management strategies for other species with similar ecological requirements and subjected to similar threats.

Endemics co-occur because they evolved in situ and persist regionally or because they evolved ex situ and later dispersed to shared habitats, generating evolutionary or ecological endemicity centres, respectively. We investigate whether different endemicity centres can intertwine in the region ranging from Alps to Sicily, by studying their butterfly fauna. We gathered an extensive occurrence data set for butterflies of the study area (27,123 records, 269 species, in cells of 0.5 × 0.5 degrees of latitude-longitude). We applied molecular-based delimitation methods (GMYC model) to 26,557 cytochrome c oxidase subunit 1 (COI) sequences of Western Palearctic butterflies. We identified entities based on molecular delimitations and/or the checklist of European butterflies and objectively attributed occurrences to their most probable entity. We obtained a zoogeographic regionalisation based on the 69 endemics of the area. Using phylogenetic ANOVA we tested if endemics from different centres differ from each other and from nonendemics for key ecological traits and divergence time. Endemicity showed high incidence in the Alps and Southern Italy. The regionalisation separated the Alps from the Italian Peninsula and Sicily. The endemics of different centres showed a high turnover and differed in phylogenetic distances, phenology and distribution traits. Endemics are on average younger than nonendemics and the Peninsula-Sicily endemics also have lower variance in divergence than those from the Alps. The observed variation identifies Alpine endemics as paleoendemics, now occupying an ecological centre, and the Peninsula-Sicily ones as neoendemics, that diverged in the region since the Pleistocene. The results challenge the common view of the Alpine-Apennine area as a single “Italian refugium”., Support for this research was provided by ‘La Caixa’ Foundation (ID 100010434) to MM (grant LCF/BQ/DR20/11790020), the Ramón y Cajal program of the Spanish Ministry of Science and Innovation to GT (RYC2018-025335-I), the Academy of Finland to VD (Academy Research Fellow, decision no. 328895), by project PID2019-107078GB-I00/AEI/10.13039/501100011033 to RV and by the projects "Ricerca e conservazione sui lepidotteri diurni di sei Parchi Nazionali dell’Appennino centro-settentrionale”, “Ricerca e conservazione sugli Impollinatori dell’Arcipelago Toscano e divulgazione sui Lepidotteri del parco” and “Servizio di attuazione delle azioni per la protezione degli impollinatori e diffusione dell’entomofauna del Parco Nazionale dell'Alta Murgia” to LD.

Satyrinae butterflies occurring in the Mediterranean apparently have reduced gene flow over sea straits, and for several species, recent wide‐scale biodiversity surveys indicate the existence of divergent mitochondrial lineages. Here, we apply an integrative approach and examine the phylogeography of the genus Lasiommata in the Western Palearctic. Our research comprised molecular analyses (mitochondrial and nuclear DNA) and geometric morphometrics (wings and genitalia) for two main species groups, and a comparative GMYC analysis, based on COI, of all the tribes within Satyrinae from this region. The GMYC approach revealed a particularly fast coalescence rate in the Parargina subtribe. The Lasiommata group was divided into 12 evolutionary significant units: six clades for the L. maera species group, five for the L. megera species group, and one for L. petropolitana, with divergences of about 1%. The patterns of COI were mirrored by ITS2 in L. maera, but the two markers were generally inconsistent in L. megera. On the contrary, morphological differences were coherent with the results of COI for L. megera, but less clearly so for L. maera. L. paramegaera and L. meadewaldoi were considerably differentiated for all the analyzed markers and likely proceeded faster in the process of speciation because of geographic isolation and reduced effective population size, rendering the rest paraphyletic. Our study illustrates the continuous nature of speciation and the difficulties of delimiting species. In Lasiommata, the recognition of taxa as diverging lineages or distinct, possibly paraphyletic species, mostly depends on the criteria adopted by different species concepts., Funding for this research came from the Spanish MINECO and AEI/FEDER, UE (CGL2013‐48277‐P and CGL2016‐76322‐P to RVi), Marie Skłodowska‐Curie Train2Move to RVo (grant 609402–2020), Marie Sklodowska‐Curie IOF grant (project 625997) to VD, from Tuscan Archipelago National Park and Florence University to LD and from the project “Barcoding‐Italian‐Butterflies.”

Glioblastoma (GBM) comprises distinct subtypes characterized by their molecular profile. Mesenchymal identity in GBM has been associated with a comparatively unfavorable prognosis, primarily due to inherent resistance of these tumors to current therapies. The identification of molecular determinants of mesenchymal transformation could potentially allow for the discovery of new therapeutic targets. Zinc Finger and BTB Domain Containing 18 (ZBTB18/ZNF238/RP58) is a zinc finger transcriptional repressor with a crucial role in brain development and neuronal differentiation. Here, ZBTB18 is primarily silenced in the mesenchymal subtype of GBM through aberrant promoter methylation. Loss of ZBTB18 contributes to the aggressive phenotype of glioblastoma through regulation of poor prognosis–associated signatures. Restitution of ZBTB18 expression reverses the phenotype and impairs tumor-forming ability. These results indicate that ZBTB18 functions as a tumor suppressor in GBM through the regulation of genes associated with phenotypically aggressive properties. Implications: This study characterizes the role of the putative tumor suppressor ZBTB18 and its regulation by promoter hypermethylation, which appears to be a common mechanism to silence ZBTB18 in the mesenchymal subtype of GBM and provides a new mechanistic opportunity to specifically target this tumor subclass. Mol Cancer Res; 15(8); 998–1011. ©2017 AACR.

Tricholapita Gómez-Zurita and Cardoso nom. nov. is proposed as the replacement name for the leaf beetle taxon Lapita Gómez-Zurita and Cardoso, 2014, nec Bickel, 2002. Moreover, the rank of Tricholapita stat. nov. is elevated from subgenus of Taophila Heller, 1916 to generic status. Phylogenetic evidence based on mtDNA rrnS sequences and diagnostic morphological characters reveals a new species from the south of Grande Terre in New Caledonia, which is described: Tricholapita reidi sp. nov.

The genus Harpactea Bristowe, 1939 (Araneae, Dysderidae) is one of the most diverse and abundant components of the Mediterranean ground-dwelling spider fauna. However, the taxonomic boundaries of the genus are unclear and its monophyly has been questioned, yet never tested, in a quantitative framework. The only taxonomic revisions in the genus trace back to the 1960s–1990s, and most subsequent work has consisted of single species descriptions. Part of the confusion surrounding the genus and species groups delimitations have to do with the lack of a standardized nomenclature for diagnostic structures based on explicit homology statements. Here we use the description of new species of Harpactea to propose a set of homologies in the male and female genitalia and a standardized nomenclature for those structures. We formally describe three new Harpactea species: H. salvatorei sp. nov., from a cave in the north-eastern Italian Alps, and H. damini sp. nov. and H. mateparlovi sp. nov. from different caves on the Biokovo Mountain and surroundings in Croatia. All the new taxa belong to the lepida species group. In addition, we use a multilocus target gene approach to provide the first hypothesis on the internal phylogenetic structure of the genus Harpactea and the subfamily Harpacteinae, which includes representatives of all currently accepted species groups in the genus and genera in the subfamily. Our results support the close affinities of the species here described but reject the monophyly of both Harpactea and some of the species groups proposed in the literature. http://zoobank.org/NomenclaturalActs/bd2739c9-c1f0-451f-88cc-06c784987a62 http://zoobank.org/NomenclaturalActs/ffb50b2e-ac7b-49a8-aa0d-408143c8a990 http://zoobank.org/NomenclaturalActs/184c58fe-6501-4cc2-ba50-cca4fcff8289., The work was financially supported by a Marie Skłodowska-Curie Individual Fellowship (749867) (MP), Agència de Gestió d'Ajuts Universitaris i de Recerca (http://agaur.gencat.cat/en/inici/), and by CGL2016-80651-P from the Spanish Ministry of Economy and Competitivity (MA) and 2017SGR83 from the Catalan Government (MA).

Understanding host plant preference and the relative quality of resource provided by co-occurring host plants is a key step to predict butterfly species abundance and responses to environmental changes, and, consequently, to plan management measures. Zerynthia cassandra is an Italian endemic species strongly dependent on the availability of its host plants, Aristolochia rotunda and Aristolochia lutea. The insular population occurring on Elba island (Tuscan Archipelago) is highly threatened, because of limited host plant distribution, small population size and apparent lack of gene flow with the mainland. In 2017, we carried out field surveys and rearing experiments to (i) identify the characteristics of the host plants (vegetative status) and the site characteristics (aspect, irradiation, distance from other patches) correlated with the number of eggs occurring on individual plants, (ii) compare larval growth, food-conversions rate and larval and adult survivorship on the two host plants species. Egg occurrence depends on patch irradiation, the number of leaves and flowers occurring on individual plants and the occurrence of nearby patches. These findings allowed to identify the optimal Aristolochia patch features for egg laying and development. Laboratory rearing success was higher than 50% and although plant species did not show a significant effect on oviposition, we found that larval and adult survival was higher on A. rotunda. Our results suggest habitat management aimed at increasing resource availability for Z. cassandra and possible ex-situ conservation actions aimed at recovering the population in case of potential catastrophic events.