DNA barcoding unveiling rare species: the case of Pruvotfolia pselliotes (Labbé, 1923) (Mollusca: Gastropoda: Nudibranchia) in the Mediterranean Sea

The Mediterranean Sea is a hot spot for marine biodiversity, and this is particularly evident taking into consideration the diversity observed in many animal groups, among them the Molluscs. In the last decade, several works have revealed a high rate of cryptic diversity characterizing the Molluscan fauna of the Mediterranean Sea and an increasing number of endemic and/or new species inhabiting this semi-enclosed basin have been recorded or described. The DNA-barcoding method is considered an essential step in the integrative taxonomy applications, to unravel cryptic diversity and for species identification. Here we report the case of DNA-barcoding technique applied to identify a nudibranch (Heterobranchia) collected from the Adriatic Sea, in the Bay of Kotor (Montenegro), for which a standard morphological identification was not possible. Mediterranean specimen belonging to Pruvotfolia pselliotes (Labbé, 1923) is for the first time molecularly identified and its COI DNA sequence compared with the one of an individual collected from the type locality. In addition, this is the first verified report of this species from the Adriatic Sea. Finally, the potential of using DNA-barcoding is here discussed, together with the habitat and the geographical distribution of this uncommon species

Isidella elongata (Cnidaria: Alcyonacea) facies in the western Mediterranean Sea: Visual surveys and descriptions of its ecological role

Isidella elongata is a candelabrum-shaped alcyonacean forming important facies on the bathyal muddy bottoms of the Mediterranean Sea, currently considered a sensitive habitat and heavily impacted by deep-sea fisheries. Until a few decades ago, this facies was a widespread habitat of the deep Mediterranean seabed and I. elongata was a common species in the trawling fishery's bycatch. Despite its current persistence in dense aggregations being very scarce, a dense facies of I. elongata was revealed during several ROV (Remotely Operated Vehicle) surveys carried out from 2010 to 2014 on the muddy bottoms between two seamounts east of Ibiza (Balearic Sea). The facies developed in an area between 480 and 615 m in depth where trawling is forbidden, with an extraordinary density of about 2300–2683 colonies/ha, representing one of the biggest facies of I. elongata currently known for the Mediterranean Sea considering the surface covered and the colonies' density. The associated community was surveyed, with 50 taxa identified. Moreover, a canyon southwest of Formentera characterised by the presence of I. elongata together with a high trawling impact was investigated. The density of the colonies was 53–62 colonies/ha and only 19 taxa of associated fauna were observed. The results of the two areas are compared and discussed in the framework of the protection of such an important habitat

New deep-water cnidarian sites in the southern Adriatic Sea

Recent ROV (Remotely Operated Vehicle) exploration and bottom sampling in the southern Adriatic Sea (Apulian and Montenegrin margins) resulted in the discovery of cnidarian-rich deep-sea habitats in the depth range of ca. 400-700 m. In particular, ROV inspection of Montenegrin canyons reveals the existence of megabenthic communities dominated by a variety of cnidarians, including scleractinians (Madrepora oculata, Lophelia pertusa, Dendrophyllia cornigera),antipatharians (Leiopathes glaberrima) and gorgonians (Callogorgia verticillata) as major habitat forming taxa, often in association with sponges and, subordinately, serpulids. All such cnidarians are new records for the south-eastern side of the Adriatic Sea. Our investigation indicates that an almost continuous belt of patchy cold water coral sites occurs along the entire south-western margin (Apulian),basically connecting the Adriatic populations with those inhabiting the Ionian margin (Santa Maria di Leuca coral province)

Transcriptional dynamics of induced pluripotent stem cell differentiation into β cells reveals full endodermal commitment and homology with human islets.

Abstract Background aims Induced pluripotent stem cells (iPSCs) have the capacity to generate β cells in vitro, but the differentiation is incomplete and generates a variable percentage of off-target cells. Single-cell RNA sequencing offers the possibility of characterizing the transcriptional dynamics throughout differentiation and determining the identity of the final differentiation product. Methods Single-cell transcriptomics data were obtained from four stages across differentiation of iPSCs into β cells and from human donor islets. Results Clustering analysis revealed that iPSCs undertake a full endoderm commitment, and the obtained endocrine pancreatic cells have high homology with mature islets. The iPSC-derived β cells were devoid of pluripotent residual cells, and the differentiation was pancreas-specific, as it did not generate ectodermal or mesodermal cells. Pseudotime trajectory identified a dichotomic endocrine/non-endocrine cell fate and distinct subgroups in the endocrine branch. Conclusions Future efforts to produce β cells from iPSCs must aim not only to improve the resulting endocrine cell but also to avoid differentiation into non-pancreatic endoderm cells

Modeling human pancreatic beta cell dedifferentiation

Objective: Dedifferentiation could explain reduced functional pancreatic β-cell mass in type 2 diabetes (T2D). Methods: Here we model human β-cell dedifferentiation using growth factor stimulation in the human β-cell line, EndoC-βH1, and human pancreatic islets. Results: Fibroblast growth factor 2 (FGF2) treatment reduced expression of β-cell markers, (INS, MAFB, SLC2A2, SLC30A8, and GCK) and activated ectopic expression of MYC, HES1, SOX9, and NEUROG3. FGF2-induced dedifferentiation was time- and dose-dependent and reversible upon wash-out. Furthermore, FGF2 treatment induced expression of TNFRSF11B, a decoy receptor for RANKL and protected β-cells against RANKL signaling. Finally, analyses of transcriptomic data revealed increased FGF2 expression in ductal, endothelial, and stellate cells in pancreas from T2D patients, whereas FGFR1, SOX,9 and HES1 expression increased in islets from T2D patients. Conclusions: We thus developed an FGF2-induced model of human β-cell dedifferentiation, identified new markers of dedifferentiation, and found evidence for increased pancreatic FGF2, FGFR1, and β-cell dedifferentiation in T2D

Insulin Storage and Glucose Homeostasis in Mice Null for the Granule Zinc Transporter ZnT8 and Studies of the Type 2 Diabetes–Associated Variants

International audienceObjective. Zinc ions are essential for the formation of hexameric insulin and hormone crystallisation. Correspondingly, a non-synonymous single nucleotide polymorphism rs13266634 in the SLC30A8 gene, encoding the secretory granule zinc transporter ZnT8, is associated with type 2 diabetes. Here, we describe the effects of deleting the ZnT8 gene in mice and explore the action of the at-risk allele. Research Design and Methods. Slc30a8 null mice were generated and backcrossed at least twice onto a C57BL/6J background. Glucose and insulin tolerance were measured by intraperitoneal injection, or euglycemic clamp, respectively. Insulin secretion, electrophysiology, imaging, and the generation of adenoviruses encoding the low- (W325) or elevated- (R325) risk ZnT8 alleles, were undertaken using standard protocols. Results. ZnT8(-/-) mice displayed age, sex and diet-dependent abnormalities in glucose tolerance, insulin secretion and body weight. Islets isolated from null mice had reduced granule zinc content, and showed age-dependent changes in granule morphology, with markedly fewer dense cores but more rod-like crystals. Glucose-stimulated insulin secretion, granule fusion and insulin crystal dissolution, as assessed by total internal reflection fluorescence microscopy, were unchanged or enhanced in ZnT8(-/-) islets. Insulin processing was normal. Molecular modelling revealed that residue-325 was located at the interface between ZnT8 monomers. Correspondingly, the R325 variant displayed lower apparent Zn(2+) transport activity than W325 ZnT8 by fluorescence-based assay. Discussion and conclusions. ZnT8 is required for normal insulin crystallisation and insulin release in vivo but not, remarkably, in vitro. Defects in the former processes in carriers of the R allele may increase type 2 diabetes risk

Benthic habitat modelling and mapping as a conservation tool for marine protected areas: A seamount in the western Mediterranean

1. An ecologically representative, well‐connected, and effectively managed system of marine protected areas (MPAs) has positive ecological and environmental effects as well as social and economic benefits. Although progress in expanding the coverage of MPAs has been made, the application of management tools has not yet been implemented in most of these areas. 2. In this work, distribution models were applied to nine benthic habitats on a Mediterranean seamount within an MPA for conservation purposes. Benthic habitat occurrences were identified from 55 remotely operated vehicle (ROV) transects, at depths from 76 to 700 m, and data derived from multibeam bathymetry. Generalized additive models (GAMs) were applied to link the presence of each benthic habitat to local environmental proxies (depth, slope, backscatter, aspect, and bathymetric position index, BPI). 3. The main environmental drivers of habitat distribution were depth, slope, and BPI. Based on this result, five different geomorphological areas were distinguished. A full coverage map indicating the potential benthic habitat distribution on the seamount was obtained to inform spatial management. 4. The distribution of those habitats identified as vulnerable marine ecosystems (VMEs) was used to make recommendations on zonation for developing the management plan of the MPA. This process reveals itself as an appropriate methodological approach that can be developed in other areas of the Natura 2000 marine networkEn prensa1,92

Zinc transporter gene expression is regulated by pro-inflammatory cytokines: a potential role for zinc transporters in beta-cell apoptosis?

<p>Abstract</p> <p>Background</p> <p>β-cells are extremely rich in zinc and zinc homeostasis is regulated by zinc transporter proteins. β-cells are sensitive to cytokines, interleukin-1β (IL-1β) has been associated with β-cell dysfunction and -death in both type 1 and type 2 diabetes. This study explores the regulation of zinc transporters following cytokine exposure.</p> <p>Methods</p> <p>The effects of cytokines IL-1β, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) on zinc transporter gene expression were measured in INS-1-cells and rat pancreatic islets. Being the more sensitive transporter, we further explored ZnT8 (Slc30A8): the effect of ZnT8 over expression on cytokine induced apoptosis was investigated as well as expression of the insulin gene and two apoptosis associated genes, BAX and BCL2.</p> <p>Results</p> <p>Our results showed a dynamic response of genes responsible for β-cell zinc homeostasis to cytokines: IL-1β down regulated a number of zinc-transporters, most strikingly ZnT8 in both islets and INS-1 cells. The effect was even more pronounced when mixing the cytokines. TNF-α had little effect on zinc transporter expression. IFN-γ down regulated a number of zinc transporters. Insulin expression was down regulated by all cytokines. ZnT8 over expressing cells were more sensitive to IL-1β induced apoptosis whereas no differences were observed with IFN-γ, TNF-α, or a mixture of cytokines.</p> <p>Conclusion</p> <p>The zinc transporting system in β-cells is influenced by the exposure to cytokines. Particularly ZnT8, which has been associated with the development of diabetes, seems to be cytokine sensitive.</p

Proteomic Analysis of Polypeptides Captured from Blood during Extracorporeal Albumin Dialysis in Patients with Cholestasis and Resistant Pruritus

Albumin dialysis using the molecular adsorbent recirculating system (MARS) is a new therapeutic approach for liver diseases. To gain insight into the mechanisms involved in albumin dialysis, we analyzed the peptides and proteins absorbed into the MARS strong anion exchange (SAX) cartridges as a result of the treatment of patients with cholestasis and resistant pruritus. Proteins extracted from the SAX MARS cartridges after patient treatment were digested with two enzymes. The resulting peptides were analyzed by multidimensional liquid chromatography coupled to tandem mass spectrometry. We identified over 1,500 peptide sequences corresponding to 144 proteins. In addition to the proteins that are present in control albumin-derived samples, this collection includes 60 proteins that were specific to samples obtained after patient treatment. Five of these proteins (neutrophil defensin 1 [HNP-1], secreted Ly-6/uPAR-related protein 1 [SLURP1], serum amyloid A, fibrinogen alpha chain and pancreatic prohormone) were confirmed to be removed by the dialysis procedure using targeted selected-reaction monitoring MS/MS. Furthermore, capture of HNP-1 and SLURP1 was also validated by Western blot. Interestingly, further analyses of SLURP1 in serum indicated that this protein was 3-fold higher in cholestatic patients than in controls. Proteins captured by MARS share certain structural and biological characteristics, and some of them have important biological functions. Therefore, their removal could be related either to therapeutic or possible adverse effects associated with albumin dialysis