Molecular Phylogeny of the Astrophorida (Porifera, Demospongiaep) Reveals an Unexpected High Level of Spicule Homoplasy

Background: The Astrophorida (Porifera, Demospongiaep) is geographically and bathymetrically widely distributed. Systema Porifera currently includes five families in this order: Ancorinidae, Calthropellidae, Geodiidae, Pachastrellidae and Thrombidae. To date, molecular phylogenetic studies including Astrophorida species are scarce and offer limited sampling. Phylogenetic relationships within this order are therefore for the most part unknown and hypotheses based on morphology largely untested. Astrophorida taxa have very diverse spicule sets that make them a model of choice to investigate spicule evolution. Methodology/Principal Findings: With a sampling of 153 specimens (9 families, 29 genera, 89 species) covering the deepand shallow-waters worldwide, this work presents the first comprehensive molecular phylogeny of the Astrophorida, using a cytochrome c oxidase subunit I (COI) gene partial sequence and the 59 end terminal part of the 28S rDNA gene (C1-D2 domains). The resulting tree suggested that i) the Astrophorida included some lithistid families and some Alectonidae species, ii) the sub-orders Euastrophorida and Streptosclerophorida were both polyphyletic, iii) the Geodiidae, the Ancorinidae and the Pachastrellidae were not monophyletic, iv) the Calthropellidae was part of the Geodiidae clade (Calthropella at least), and finally that v) many genera were polyphyletic (Ecionemia, Erylus, Poecillastra, Penares, Rhabdastrella, Stelletta and Vulcanella). Conclusion: The Astrophorida is a larger order than previously considered, comprising ca. 820 species. Based on these results, we propose new classifications for the Astrophorida using both the classical rank-based nomenclature (i.e., Linnaean classification) and the phylogenetic nomenclature following the PhyloCode, independent of taxonomic rank. A key to the Astrophorida families, sub-families and genera incertae sedis is also included. Incongruences between our molecular tree and the current classification can be explained by the banality of convergent evolution and secondary loss in spicule evolution. These processes have taken place many times, in all the major clades, for megascleres and microscleres

Unique spicules may confound species differentiation: Taxonomy and biogeography of Melonanchora Carter, 1874 and two new related genera (Myxillidae: Poecilosclerida) from the Okhotsk Sea

Sponges are amongst the most difficult benthic taxa to properly identify, which has led to a prevalence of cryptic species in several sponge genera, especially in those with simple skeletons. This is particularly true for sponges living in remote or hardly accessible environments, such as the deep-sea, as the inaccessibility of their habitat and the lack of accurate descriptions usually leads to misclassifications. However, species can also remain hidden even when they belong to genera that have particularly characteristic features. In these cases, researchers inevitably pay attention to these peculiar features, sometimes disregarding small differences in the other “typical” spicules. The genus Melonanchora Carter, 1874, is among those well suited for a revision, as their representatives possess a unique type of spicule (spherancorae). After a thorough review of the material available for this genus from several institutions, four new species of Melonanchora, M. tumultuosa sp. nov., M. insulsa sp. nov., M. intermedia sp. nov. and M. maeli sp. nov. are formally described from different localities across the Atlanto-Mediterranean region. Additionally, all Melonanchora from the Okhotsk Sea and nearby areas are reassigned to other genera; Melonanchora kobjakovae is transferred to Myxilla (Burtonanchora) while two new genera, Hanstoreia gen. nov. and Arhythmata gen. nov. are created to accommodate Melonanchora globogilva and Melonanchora tetradedritifera, respectively. Hanstoreia gen. nov. is closest to Melonanchora, whereas Arhythmata gen. nov., is closer to Stelodoryx, which is most likely polyphyletic and in need of revision.publishedVersio

Macro and microstructural characteristics of north Atlantic deep-sea sponges as bioinspired models for tissue engineering scaffolding

Sponges occur ubiquitously in the marine realm and in some deep-sea areas they dominate the benthic communities forming complex biogenic habitats â sponge grounds, aggregations, gardens and reefs. However, deep-sea sponges and spongegrounds are still poorly investigated with regards to biotechnological potential in support of a Blue growth strategy. Under the scope of this study, five dominant North Atlantic deep-sea sponges, were characterized to elucidate promising applications in human health, namely for bone tissue engineering approaches. Geodia barretti (Gb), Geodia atlantica (Ga), Stelletta normani (Sn), Phakellia ventilabrum (Pv), and Axinella infundibuliformis (Ai), were morphologically characterized to assess macro and microstructural features, as well as chemical composition of the skeletons, using optical and scanning electron microscopy, energy dispersive x-ray spectroscopy and microcomputed tomography analyses. Moreover, compress tests were conducted to determine the mechanical properties of the skeletons. Results showed that all studied sponges have porous skeletons with porosity higher than 68%, pore size superior than 149 mm and higher interconnectivity (>96%), thus providing interesting models for the development of scaffolds for tissue engineering. Besides that, EDS analyses revealed that the chemical composition of sponges, pointed that demosponge skeletons are mainly constituted by carbon, silicon, sulfur, and oxygen combined mutually with organic and inorganic elements embedded its internal architecture that can be important features for promoting bone matrix quality and bone mineralization. Finally, the morphological, mechanical, and chemical characteristics here investigated unraveled the potential of deep-sea sponges as a source of biomaterials and biomimetic models envisaging tissue engineering applications for bone regeneration.The authors would like to acknowledge the funding from the European Union Framework Program for Research and Innovation Horizon 2020 through project SponGES (H2020- BG-01-2015-679849) and from the Northern Portugal Regional Operational Program (NORTE2020), under the Portugal 2020 Partnership Agreement, through the Structured projects for R&D&I NORTE-01-0145-FEDER-000021 and NORTE-01-0145- FEDER-000023. JRX research was further supported by national funds through FCT Foundation for Science and Technology within the scope of UIDB/04423/2020 and UIDP/04423/2020, and CEECIND/00577/2018

Análise in vitro da distribuição de tensões em implantes angulados de diferentes comprimentos

: Neste trabalho apresenta-se um estudo realizado para avaliar computacionalmente ouso de implantes curtos em reabilitação mandibular total com base no Método dos ElementosFinitos. Em particular, verifica-se como o comprimento desses implantes influencia adistribuição de tensões durante a aplicação de cargas mastigatórias em reabilitaçõesmandibulares de acordo com o conceito All-on-4®

Glass sponge grounds on the Scotian Shelf and their associated biodiversity

Emerald Basin on the Scotian Shelf off Nova Scotia, Canada, is home to a globally unique population of the glass sponge Vazella pourtalesi. Through the analysis of both in situ photographs and trawl catch data from annual multispecies bottom-trawl surveys, we examined community composition, species density, and abundance of epibenthos and fish associated with V. pourtalesi compared to locations without this sponge. Using generalized linear models and analysis of similarities, the importance of V. pourtalesi in enhancing species density and abundance of the associated epibenthic community was assessed against that of the hard substrate on which it settles. Our results indicated that the megafaunal assemblage associated with V. pourtalesi was significantly different in composition and higher in species density and abundance compared to locations without V. pourtalesi. Analysis of similarity of trawl catch data indicated that fish communities associated with the sponge grounds are significantly different from those without V. pourtalesi, although no species were found exclusively on the sponge grounds. Our study provides further evidence of the role played by sponge grounds in shaping community structure and biodiversity of associated deep-sea epibenthic and fish communities. The mechanism for biodiversity enhancement within the sponge grounds formed by V. pourtalesi is likely the combined effect of both the sponge itself and its attachment substrate, which together comprise the habitat of the sponge grounds. We also discuss the role of habitat provision between the mixed-species tetractinellid sponges of the Flemish Cap and the monospecific glass sponge grounds of Emerald Basin.publishedVersio

Genetic diversity, connectivity and gene flow along the distribution of the emblematic Atlanto-Mediterranean sponge Petrosia ficiformis (Haplosclerida, Demospongiae)

Background: Knowledge about the distribution of the genetic variation of marine species is fundamental to address species conservation and management strategies, especially in scenarios with mass mortalities. In the Mediterranean Sea, Petrosia ficiformis is one of the species most affected by temperature-related diseases. Our study aimed to assess its genetic structure, connectivity, and bottleneck signatures to understand its evolutionary history and to provide information to help design conservation strategies of sessile marine invertebrates. Results: We genotyped 280 individuals from 19 locations across the entire distribution range of P. ficiformis in the Atlanto-Mediterranean region at 10 microsatellite loci. High levels of inbreeding were detected in most locations (especially in the Macaronesia and the Western Mediterranean) and bottleneck signatures were only detected in Mediterranean populations, although not coinciding entirely with those with reported die-offs. We detected strong significant population differentiation, with the Atlantic populations being the most genetically isolated, and show that six clusters explained the genetic structure along the distribution range of this sponge. Although we detected a pattern of isolation by distance in P. ficiformis when all locations were analyzed together, stratified Mantel tests revealed that other factors could be playing a more prominent role than isolation by distance. Indeed, we detected a strong effect of oceanographic barriers impeding the gene flow among certain areas, the strongest one being the Almeria-Oran front, hampering gene flow between the Atlantic Ocean and the Mediterranean Sea. Finally, migration and genetic diversity distribution analyses suggest a Mediterranean origin for the species. Conclusions: In our study Petrosia ficiformis showed extreme levels of inbreeding and population differentiation, which could all be linked to the poor swimming abilities of the larva. However, the observed moderate migration patterns are highly difficult to reconcile with such poor larval dispersal, and suggest that, although unlikely, dispersal may also be achieved in the gamete phase. Overall, because of the high genetic diversity in the Eastern Mediterranean and frequent mass mortalities in the Western Mediterranean, we suggest that conservation efforts should be carried out specifically in those areas of the Mediterranean to safeguard the genetic diversity of the species

Bioactivity of biosilica obtained from North Atlantic deep-sea sponges

Demosponges are a well-known source of a plethora of bioactive compounds. In particular, they are able to form a skeleton by direct deposition of silica in a process catalysed by silicatein. Herein, we isolated biosilicas from five different Atlantic deep-sea sponges Geodia atlantica (GA), Geodia barretti (GB), Stelletta normani (SN), Axinella infundibuliformis (AI) and Phakellia ventilabrum (PV) to explore the bioactivity and osteogenic capacity of its silica-based materials. We chemically characterized the isolated biosilicas and evaluated them for their bioactivity to deposit Ca and P on their surface (by immersion in simulated body fluid, SBF). GB-, SN-, AI- and PV-based biosilicas did not generate a stable calcium phosphate (CaP) layer over time in the presence of SBF, however, the GA-derived one was able to form a CaP surface layer (at a Ca/P ratio of ~1.7, similar to the one observed for hydroxyapatite), that was stable during the 28 days of testing. In addition, no cytotoxicity towards L929 and SaOs2 cells was observed for the GA-based biosilica up to a concentration of 10 mg/mL. Overall, the GA-based biosilica presents the characteristics to be used in the development of biomaterials for bone tissue engineering (BTE).This research has been performed with funding from the European Union’s Horizon 2020 research and innovation programme under Grant Agreements No. 679849 (SponGES) and No. 668983 (FORECAST). Funding from the European Regional Development Fund, through Atlantic Area Program, under the scope of BlueHuman project (EAPA_151/2016) is also acknowledged. JRX research was also supported by National Funds through FCT, the Portuguese Foundation for Science and Technology within the scope of UIDB/04423/2020, UIDP/04423/2020, and CEECIND/00577/2018

Targeted therapy using phage technology: a computational and experimental breast cancer study

During the past two decades cancer biology knowledge has widely increased and shifted the paradigm of cancer treatment from nonspecific cytotoxic agents to selective, mechanism-based therapeutics. Initially, cancer drug design was focused on compounds that rapidly killed dividing cells. Though still used as the backbone of current treatments, these highly unspecific targeting drugs lead to significant toxicity for patients, narrowing the therapeutic index, and frequently lead to drug resistance. Therefore, cancer therapies are now based on cancer immunotherapy and targeted agents, whereas novel treatments are strategically combining both to improve clinical outcomes. Despite the nanotechnology advances dictating the development of targeted therapies in diverse classes of nano-based carriers, virus-based vectors still remain highly used due to its biocompatibility and specificity for the target. Particularly, bacteriophages are an interesting alternative ‘nanomedicine’ that can combine biological and chemical components into the same drug delivery system. The great potential of this novel platform for cancer therapy is the ability to genetically manipulate the virus-vector to display specific targeting moieties. Phage display technology, a general technique used for detecting interfaces of various types of interacting proteins outside of the immunological context, allows the target agents to locate the target (with an increased selection process for the specific binding – termed biopanning) and play their essential role inhibiting molecular pathways crucial for tumour growth and maintenance. Phage display specificity core is related with the binding of small peptides displayed at their coat or capsid proteins, enriched during biopanning. Bioinformatics plays an important role in testing and improving phage display libraries by effective epitope mapping, selecting from a large set of random peptides those with a high binding affinity to a target of interest. In this work we demonstrate the screening of a manually constructed 7-mer peptide library of M13KE phage particles against MDA-MB-231 and -435 cancer cell lines. Two peptides – TLATVEV and PRLNVSP – with high affinity for the referred cells were identified, respectively. Based on computationally predicted epitopes based on the peptides extracted from this library the linear peptide sequence was docked onto known membrane proteins from the used cell lines and peptides-proteins interactions were mapped. Umbrella sampling studies were performed to predict the binding affinity and to improve future rational design of binding peptides to these cancer cells

Getting Value from Pulp and Paper Industry Wastes: On the Way to Sustainability and Circular Economy

The pulp and paper industry is recognized as a well-established sector, which throughout its process, generates a vast amount of waste streams with the capacity to be valorized. Typically, these residues are burned for energy purposes, but their use as substrates for biological processes could be a more efficient and sustainable alternative. With this aim, it is essential to identify and characterize each type of waste to determine its biotechnological potential. In this context, this research highlights possible alternatives with lower environmental impact and higher revenues. The bio-based pathway should be a promising alternative for the valorization of pulp and paper industry wastes, in particular for bioproduct production such as bioethanol, polyhydroxyalkanoates (PHA), and biogas. This article focuses on state of the art regarding the identification and characterization of these wastes, their main applied deconstruction technologies and the valorization pathways reported for the production of the abovementioned bioproductspublishe