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To ensure environmental and public safety, critical facilities require rigorous seismic hazard analysis to define seismic input for their design. We consider the case of the Trans Adriatic Pipeline (TAP), which is a pipeline that transports natural gas from the Caspian Sea to southern Italy, crossing active faults and areas characterized by high seismicity levels. For this pipeline, we develop a Probabilistic Seismic Hazard Assessment (PSHA) for the broader area, and, for the selected critical sites, we perform deterministic seismic hazard assessment (DSHA), by calculating shaking scenarios that account for the physics of the source, propagation, and site effects. This paper presents a DSHA for a compressor station located at Fier, along the Albanian coastal region. Considering the location of the most hazardous faults in the study site, revealed by the PSHA disaggregation, we model the ground motion for two different scenarios to simulate the worst-case scenario for this compressor station. We compute broadband waveforms for receivers on soft soils by applying specific transfer functions estimated from the available geotechnical data for the Fier area. The simulations reproduce the variability observed in the ground motion recorded in the near-earthquake source. The vertical ground motion is strong for receivers placed above the rupture areas and should not be ignored in seismic designs; furthermore, our vertical simulations reproduce the displacement and the static offset of the ground motion highlighted in recent studies. This observation confirms the importance of the DSHA analysis in defining the expected pipeline damage functions and permanent soil deformations.

The design of critical facilities needs a targeted computation of the expected ground motion levels. The Trans Adriatic Pipeline (TAP) is the pipeline that transports natural gas from the Greek-Turkish border, through Greece and Albania, to Italy. We present here the probabilistic seismic hazard analysis (PSHA) that we performed for this facility, and the deaggregation of the results, aiming to identify the dominant seismic sources for a selected site along the Albanian coast, where one of the two main compressor stations is located. PSHA is based on an articulated logic tree of twenty branches, consisting of two models for source, seismicity, estimation of the maximum magnitude, and ground motion. The area with the highest hazard occurs along the Adriatic coast of Albania (PGA between 0.8 and 0.9 g on rock for a return period of 2475 years), while strong ground motions are also expected to the north of Thessaloniki, Kavala, in the southern Alexandroupolis area, as well as at the border between Greece and Turkey. The earthquakes contributing most to the hazard of the test site at high and low frequencies (1 and 5 Hz) and the corresponding design events for the TAP infrastructure have been identified as local quakes with MW 6.6 and 6.0, respectively.

One of the biggest challenges of the COVID-19 pandemic is the heterogeneity in disease severity exhibited amongst patients. Among multiple factors, latest studies suggest vitamin D deficiency and pre-existing health conditions to be major contributors to death from COVID-19. It is known that certain urban form attributes can impact sun exposure and vitamin D synthesis. Also, long-term exposure to air pollution can play an independent role in vitamin D deficiency. We conducted a correlational analysis of urban form and air quality in relation to the demographics and COVID-19 incidence and mortality across 32 London boroughs between March 2020 and January 2021. We found total population, number of residents of Asian ethnicity, 4-year average PM10 levels and road length to be positively correlated with COVID-19 cases and deaths. We also found percentage of households with access to total open space to be negatively correlated with COVID-19 deaths. Our findings link COVID-19 incidence and mortality across London with environmental variables linked to vitamin D status. Our study is entirely based on publicly available data and provides a reference framework for further research as more data are gathered and the syndemic dimension of COVID-19 becomes increasingly relevant in connection to health inequalities within large urban areas.

Background/objectives Epidemiological evidence indicates obesity in childhood and adolescence to be an independent risk factor for cancer and premature mortality in adulthood. Pathological implications from excess adiposity may begin early in life. Obesity is concurrent with a state of chronic inflammation, a well-known aetiological factor for DNA damage. In addition, obesity has been associated with micro-nutritional deficiencies. Vitamin D has attracted attention for its anti-inflammatory properties and role in genomic integrity and stability. The aim of this study was to determine a novel approach for predicting genomic instability via the combined assessment of adiposity, DNA damage, systemic inflammation, and vitamin D status. Subjects/methods We carried out a cross-sectional study with 132 participants, aged 10–18, recruited from schools and paediatric obesity clinics in London. Anthropometric assessments included BMI Z-score, waist and hip circumference, and body fat percentage via bioelectrical impedance. Inflammation and vitamin D levels in saliva were assessed by enzyme-linked immunosorbent assay. Oxidative DNA damage was determined via quantification of 8-hydroxy-2′-deoxyguanosine in urine. Exfoliated cells from the oral cavity were scored for genomic instability via the buccal cytome assay. Results As expected, comparisons between participants with obesity and normal range BMI showed significant differences in anthropometric measures (p p p p p p p p Conclusions Obesity, oxidative DNA damage, and vitamin D deficiency are significant predictors of genomic instability. Non-invasive biomonitoring and predictive modelling of genomic instability in young patients with obesity may contribute to the prioritisation and severity of clinical intervention measures.

A population of more than six million people worldwide at high risk of Alzheimer’s disease (AD) are those with Down Syndrome (DS, caused by trisomy 21 (T21)), 70% of whom develop dementia during lifetime, caused by an extra copy of β-amyloid-(Aβ)-precursor-protein gene. We report AD-like pathology in cerebral organoids grown in vitro from non-invasively sampled strands of hair from 71% of DS donors. The pathology consisted of extracellular diffuse and fibrillar Aβ deposits, hyperphosphorylated/pathologically conformed Tau, and premature neuronal loss. Presence/absence of AD-like pathology was donor-specific (reproducible between individual organoids/iPSC lines/experiments). Pathology could be triggered in pathology-negative T21 organoids by CRISPR/Cas9-mediated elimination of the third copy of chromosome 21 gene BACE2, but prevented by combined chemical β and γ-secretase inhibition. We found that T21 organoids secrete increased proportions of Aβ-preventing (Aβ1–19) and Aβ-degradation products (Aβ1–20 and Aβ1–34). We show these profiles mirror in cerebrospinal fluid of people with DS. We demonstrate that this protective mechanism is mediated by BACE2-trisomy and cross-inhibited by clinically trialled BACE1 inhibitors. Combined, our data prove the physiological role of BACE2 as a dose-sensitive AD-suppressor gene, potentially explaining the dementia delay in ~30% of people with DS. We also show that DS cerebral organoids could be explored as pre-morbid AD-risk population detector and a system for hypothesis-free drug screens as well as identification of natural suppressor genes for neurodegenerative diseases.

En este trabajo de investigación se estudia una alternativa de reutilización de materiales de desecho industrial en concreto reforzado buscando determinar parámetros experimentales de resistencia a la corrosión de los distintos sistemas de concreto, en comparación a un concreto reforzado convencional. El uso de la técnica de espectroscopia de impedancia electroquímica muestra que la interacción del jal como cementante no modifica el proceso de corrosión, pues se observa que el uso de residuo minero hasta un 10% no favorece la llegada de los agentes corrosivos a la superficie de la varilla de refuerzo en el concreto comparado a un sistema convencional. Los valores de Rp son muy similares para los distintos sistemas. La interacción de cloruros con el metal se ve aumentada al usar 15% en la dosificación de jal como cementante, modificando considerablemente la velocidad de corrosión del sistema. El concreto elaborado con jal como cementante tiene un comportamiento electroquímico y mecánico muy similar a un concreto convencional hasta una dosificación de 10%.

// Tiziana Negri 1,* , Silvia Brich 1,2,* , Fabio Bozzi 1 , Chiara V. Volpi 3 , Ambra V. Gualeni 3 , Silvia Stacchiotti 4 , Loris De Cecco 5 , Silvana Canevari 5 , Annunziata Gloghini 3 and Silvana Pilotti 1 1 Department of Diagnostic Pathology and Laboratory Medicine, Laboratory of Experimental Molecular Pathology, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy 2 MOSE-DEA, University of Trieste, Trieste, Italy 3 Department of Diagnostic Pathology and Laboratory Medicine, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy 4 Adult Mesenchymal Tumor and Rare Cancer Medical Oncology Unit, Cancer Medicine Department, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy 5 Department of Experimental Oncology and Molecular Medicine, Functional Genomics and Bioinformatics, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy * These authors have contributed equally to the study Correspondence to: Silvana Pilotti, email: // Keywords : GEP; MErT/EMT; immunological ignorance; AR; Meflin/PDGFRA Received : February 08, 2017 Accepted : March 13, 2017 Published : March 22, 2017 Abstract To gain new insights into desmoplastic small round cell tumors (DSRCTs) by means of gene expression profiling (GEP). Formalin-fixed, paraffin-embedded surgical specimens obtained from seven pretreated DSRCT patients were interrogated using GEP complemented by immunohistochemistry, a cancer stem cell array, and miRNA in situ hybridisation, including the combined chimera modules miRNA-200/ZEB1 and miRNA-34/SLUG. The chimera modules divided the cases into three classes that respectively recapitulated the traits of mesenchymal epithelial reverse transition (MErT), epithelial mesenchymal transition (EMT), and hybrid/partial EMT. This indicates a close correlation between the reprogramming governed by EMT regulators and DSRCT biology, which was further confirmed by miRNA-21 and is consistent with the broad morphological spectrum of DSRCTs. Starting from the miRNA-200/ZEB1 axis, we also found that DSRCTs carry a signature of immunological ignorance that is not responsive to PD-­L1 blockade. Evidence that the up-regulation of miRNA-200 and E-cadherin, and quite a high level of miRNA-21 expression segregate with the MErT supports the idea that, in addition to the hybrid/partial state, MErT is also enriched in stemness: the androgen-positive cases, whose stemness traits were confirmed by stem cell arrays, all fell into these two classes. Our findings also confirmed that tumoral cell PDGFRA expression correlates with desmoplasia, and demonstrated the co-expression of PDGFRA and ISLR/Meflin, another marker of pluripotency. Despite the limited number of cases, these findings provide unexpectedly relevant information concerning the pathogenesis of DSRCTs, and prove the validity of miRNA-based chimera circuit modelling in the clinico-pathological setting.

A population of >6 million people worldwide at high risk of Alzheimer’s disease (AD) are those with Down Syndrome (DS, caused by trisomy 21 (T21)), 70% of whom develop dementia during lifetime, caused by an extra copy of β-amyloid-(Aβ)-precursor-protein gene. We report AD-like pathology in cerebral organoids grownin vitrofrom non-invasively sampled strands of hair from 71% of DS donors. The pathology consisted of extracellular diffuse and fibrillar Aβ deposits, hyperphosphorylated/pathologically conformed Tau, and premature neuronal loss.Presence/absence of AD-like pathology was donor-specific (reproducible between individual organoids/iPSC lines/experiments). Pathology could be triggered in pathology-negative T21 organoids by CRISPR/Cas9-mediated elimination of the third copy of chromosome-21-geneBACE2, but prevented by combined chemical β and γ-secretase inhibition. We found that T21-organoids secrete increased proportions of Aβ-preventing (Aβ1-19) and Aβ-degradation products (Aβ1-20 and Aβ1-34). We show these profiles mirror in cerebrospinal fluid of people with DS. We demonstrate that this protective mechanism is mediated by BACE2-trisomy and cross-inhibited by clinically trialled BACE1-inhibitors. Combined, our data prove the physiological role ofBACE2as a dose-sensitive AD-suppressor gene, potentially explaining the dementia delay in ∼30% of people with DS. We also show that DS cerebral organoids could be explored as pre-morbid AD-risk population detector and a system for hypothesis-free drug screens as well as identification of natural suppressor genes for neurodegenerative diseases.

Purpose: Recent studies have raised concerns that obese children may present an increased pre-disposition towards age–related disorders such as cancer. Initial evidence would suggest that the increased risk of developing cancer later in life might be linked to the reported presence of chronic low–grade inflammation in childhood obesity and the sustained accumulation of Reactive Oxygen Species (ROS) capable of inflicting DNA damage, a well–known promoter and driver of carcinogenesis. This conceptual framework outlines the clinical and laboratory investigations required for the combined assessment and monitoring of systemic inflammation, micro–nutritional deficiencies and acquired genome damage in childhood obesity. Approach: A case–control study is proposed in a cohort of obese and healthy weight 11–15 year olds. This study combines assessments of blood, together with minimally invasive investigations of saliva and urine samples to obtain an overview of inflammation and DNA damage status in obesity. Furthermore, we present a nutrition sensitive intervention programme to investigate the reversibility of these pathological states. Findings: It is hypothesised that chronic inflammation in childhood obesity can establish a harmful microenvironment that inflicts damage to DNA. It is postulated that a nutrition sensitive intervention may reverse implications of inflammation and DNA damage in obese children. Value: An evaluation of the role of genomic instability and cumulative DNA damage early in life as a possible causative link between childhood obesity and the ncreased risk of developing cancer in adulthood. The conceptual framework proposes a personalised approach to disease diagnosis and monitoring of ‘genome health’ that can inform the prioritisation and severity of intervention measures in the clinical setting.

The interpretation of approximately 1300 km of multichannel seismic data, acquired in the northern Adriatic Sea, revealed that the Apennine foredeep in the northern Adriatic does not show any evidence of tilting since Early Pliocene, suggesting that slab retreat cannot be the geodynamic driving force for the northern Adriatic subsidence and for the northern Apennine kinematic since that time. Moreover, the presence of two fault systems that developed since the Pliocene indicates that the northern Adriatic underwent intraplate deformation during the Plio-Pleistocene. These considerations suggest that, since Early Pliocene, subduction below the northern Apennine ceased, and that the Adria Plate movements were mainly driven by the kinematics of the adjacent plates, particularly by the Nubia-Europe convergence. At present, on the basis of the foredeep setting, four distinct sectors can be recognized in the Adriatic foreland: Po Plain, northern, central and southern Adriatic, separated by major tectonic lineaments. The northern Adriatic differs from the two adjoining sectors because its foredeep deposits are still well preserved. In the Po Plain in fact, the Adriatic foredeep is almost totally incorporated into the Alpine and Apennine chains while in the central Adriatic, it is affected by a considerable intraplate deformation (e.g. the Mid Adriatic Ridge). In this framework, the northern Adriatic region may be regarded as a triangular shaped sector indented below the Southern Alps to the north and the northern Dinarides to the north-east and bounded to the west by the Schio-Vicenza fault system and to the south-east by the Kvarner fault.

Epidemiological evidence linking obesity with increased risk of cancer is steadily growing, although the causative aspects underpinning this association are only partially understood. Obesity leads to a physiological imbalance in the regulation of adipose tissue and its normal functioning, resulting in hyperglycaemia, dyslipidaemia and inflammation. These states promote the generation of oxidative stress, which is exacerbated in obesity by a decline in anti-oxidant defence systems. Oxidative stress can have a marked impact on DNA, producing mutagenic lesions that could prove carcinogenic. Here we review the current evidence for genomic instability, sustained DNA damage and accelerated genome ageing in obesity. We explore the notion of genotoxicity, ensuing from systemic oxidative stress, as a key oncogenic factor in obesity. Finally, we advocate for early, pre-malignant assessment of genome integrity and stability to inform surveillance strategies and interventions.

INTRODUCTION Scientists are building detailed maps of the cellular composition in the human brain to learn about its development. In the human cortex, the largest area of the mammalian brain, neural circuits are formed through anatomical refinement, including axon and synaptic pruning, and the emergence of complex patterns of network activity during early fetal development. Cellular analyses in the human brain are restricted to postmortem material, which cannot reveal the process of development. Model organisms are, therefore, commonly used for studies of brain physiology, development, and pathogenesis, but the results from model organisms do not always translate to humans. RATIONALE Systems to model human neuron dynamics and their dysfunction in vivo are needed. While biopsy specimens and the generation of neurons from induced pluripotent stem cells (iPSCs) could provide the necessary human genetic background, two- and three-dimensional cultures lack factors that normally support neuronal development, including blood vessels, immune cells, and interaction with innervating neurons from other brain areas. On the basis of previous stem cell transplantation studies in mice, we reasoned that the physiological microenvironment of the adult mouse brain could support the growth of human cortical tissue grafts that had been generated from iPSC-derived neuronal progenitors. With human neurons implanted into the mouse brain, high-resolution, real-time in vivo monitoring of human neuron dynamics for periods of time spanning the range from subseconds to several months becomes feasible. RESULTS We found that transplanted human iPSC–derived neuronal progenitors consistently assembled into vascularized territories with complex cytoarchitecture, mimicking key features of the human fetal cortex, such as its large size and cell diversification. Single-cell-resolution intravital microscopy showed that human neuronal arbors were refined via branch-specific retraction, rather than degeneration. Human synaptic networks restructured over the course of 4 months, while maintaining balanced rates of synapse formation and elimination. Human functional neurons rapidly and consistently acquired oscillatory population activity, which persisted over the 5-month observation period. Lastly, we used cortical tissue grafts derived from the fibroblasts of two individuals with Down syndrome, caused by supernumerary chromosome 21. We found that neuronal synapses in cells derived from these individuals were overly stable and that oscillatory neural activity was reduced in these grafts, revealing in vivo cellular phenotypes not otherwise apparent. CONCLUSION By combining live imaging in a multistructured tissue environment in mice with a human-specific genetic background, we provide insights into the earliest stages of human axon, synaptic, and network activity development and uncover cellular phenotypes in Down syndrome. Our work provides an alternative experimental system that can be used to study other disorders affecting the developing human cortex.

International audience; In this paper, we present an overview of the SiteChar workflow model for sitecharacterisation and assessment for CO2 storage. Site characterisation and assessment is requiredwhen permits are requested from the legal authorities in the process of starting a CO2 storageprocess at a given site. The goal is to assess whether a proposed CO2 storage site can indeed beused for permanent storage while meeting the safety requirements demanded by the EuropeanCommission (EC) Storage Directive (EU, 2009, Storage Directive 2009/31/EC). Many issueshave to be scrutinised, and the workflow presented here is put forward to help efficiently organisethis complex task.Three issues are highlighted: communication within the working team and with the authorities;interdependencies in the workflow and feedback loops; and the risk-based character of theworkflow. A general overview (helicopter view) of the workflow is given; the issues involved incommunication and the risk assessment process are described in more detail. The workflow asdescribed has been tested within the SiteChar project on five potential storage sites throughoutEurope. This resulted in a list of key aspects of site characterisation which can help prepare andfocus new site characterisation studies.

The Southern Adriatic Sea is one of the five prospective areas for CO2 storage being evaluated under the FP7 European Sitechar project. The potential reservoir identified in the investigated area is represented by a carbonate formation (Scaglia Formation – Late Cretaceous). This paper shows the site characterization applied to one of the structures identified in the carbonate storage system of the South Adriatic offshore. The interpretation and analysis of seismic and borehole data allowed the construction of a 3D geological static model on both regional and local scales. Dynamic modeling was applied, adopting a sensitivity approach (i.e. fault transmissivity, petrophysical properties of the caprock and reservoir, and different stress regimes). Coupled fluid flow and geomechanical simulation was applied to investigate the potential risk of leakage induced by mechanical solicitation on the faults occurring in the investigated area.

In the Northern Adriatic Sea, the occurrence of gas seepage and of unique rock outcrops has been widely documented. The genesis of these deposits has recently been ascribed to gas venting, leading to their classification as methane-derived carbonates. However, the origin of seeping gas was not clearly constrained. Geophysical data collected in 2009 reveal that the gas-enriched fluid vents are deeply rooted. In fact, the entire Plio-Quaternary succession is characterized by widespread seismic anomalies represented by wipe-out zones, and interpreted as gas chimneys. They commonly root at the base of the Pliocene sequence but also within the Palaeogene succession, where they appear to be associated to deep-seated faults. We suggest that there is a structural control on chimney distribution. Chimneys originate and terminate at different stratigraphic levels; commonly they reach the seafloor, where authigenic carbonate deposits form locally. Gas analyses of some gas bubble streams just above the rock outcrops reveal that gas is composed mainly of methane. Geochemical analyses performed at four selected outcrop sites show that these deposits formed as a consequence of active gas venting. In particular, geochemical analyses indicate carbonate precipitation from microbial oxidation of methane-rich fluids, although a straightforward correlation with the source depth of gas feeding the authigenic carbonates cannot yet be clearly defined.

The present work describes the results of a modeling study addressing the geological sequestration of carbon dioxide (CO 2 ) in an offshore multi-compartment reservoir located in Italy. The study is part of a large scale project aimed at implementing carbon capture and storage (CCS) technology in a power plant in Italy within the framework of the European Energy Programme for Recovery (EEPR). The processes modeled include multiphase flow and geomechanical effects occurring in the storage formation and the sealing layers, along with near wellbore effects, fault/thrust reactivation and land surface stability, for a CO 2 injection rate of 1 × 10 6 ton/a. Based on an accurate reproduction of the three-dimensional geological setting of the selected structure, two scenarios are discussed depending on a different distribution of the petrophysical properties of the formation used for injection, namely porosity and permeability. The numerical results help clarify the importance of: (i) facies models at the reservoir scale, properly conditioned on wellbore logs, in assessing the CO 2 storage capacity; (ii) coupled wellbore-reservoir flow in allocating injection fluxes among permeable levels; and (iii) geomechanical processes, especially shear failure, in constraining the sustainable pressure buildup of a faulted reservoir.

Carbonate successions have a large distribution in the Italian territory, both onshore and offshore, and they are commonly hydrocarbons exploration targets. However, an evaluation of the suitability of these sedimentary successions for CCS projects has never been performed. The present study has allowed for the first time the recognition and the geological and structural characterization of areas potentially suitable for CO2 geological storage in carbonate rocks in Italy. To achieve this objective, public available well data and 2D multichannel seismic profiles were analyzed and interpreted, allowing to define eight areas (Malossa–San Bartolomeo, Lachiarella–Binasco, Abruzzi offshore, Abruzzo–Molise, Southern Adriatic, Northern Bradanic Trough, Southern Bradanic Trough, Sicily Channel) characterized by potential reservoirs in carbonate rocks sealed by relatively thick and laterally continuous caprocks. The present results have also highlighted marked spatial heterogeneities in the recognized reservoirs, inferred to result from primary depositional processes, diagenesis and fracturing due to tectonic events. Most reservoirs were recognized within shallow marine carbonate platform succession, among which the most suitable for CCS purposes are those composed of dolostones, showing maximum porosity and permeability values of 25% and 400–450 mD, respectively. Among the identified areas, the so called “Sicily Channel” and “Abruzzi offshore” are probably those most promising because of their location in offshore settings, closeness to significant sources of CO2 and distance from the main seismogenetic tectonic structures, as well as for the occurrence of one or multiple reservoir–caprock systems. However, the information provided by this study is at a regional scale, and therefore more detailed analyses are needed to identify and characterize potential individual storage sites for CCS projects.

In 2009, the National Institute of Oceanography and Experimental Geophysics (OGS, Italy) has performed a geophysical survey in the northernmost sector of the Northern Adriatic Sea, between the Tagliamento and Po river deltas, with R/V OGS Explora. About 820 km of 2D multichannel seismic and Chirp profiles, together with multibeam data along the ship tracks, have been acquired, with the aim to reconstructing the Plio-Quaternary stratigraphic and tectonic setting of the study area. Data have been also analyzed to verify whether this region is suitable for CO2 geological storage. Sites eligible for the application of Carbon Capture and Storage (CCS) techniques have been already identified in the Adriatic Sea, except in the northernmost sector, due to the scarcity of available data. The analyses of the new OGS seismic dataset highlighted the occurrence of peculiar seismic anomalies, represented by sub-vertical wipe-out zones, that have been interpreted as due to gas leakages affecting the Plio-Quaternary sequence. They could be possibly related to the formation of rock outcrops interpreted as methane-derived carbonates. It is suggested that gas migrates along sub-vertical chimneys throughout the Plio-Quaternary sequence. The correlation of these sub-vertical features among the seismic profiles reveals that two main alignments could be recognized: a NW–SE trend offshore the Venice Lagoon and a NE–SW in the northernmost part of the investigated area. It leads to hypothesize that the preferential conduits for gas migration are associated to tectonic lineaments. The analysis of the tectonic setting has been then addressed at defining the role of fault/fracture zones, which can affect the integrity of a CO2 storage complex in this area. Tectonic features have been identified in the Mesozoic–Paleogene succession, but their relationship with the shallow faults/fractures representing the paths for gas leakages still need further work and additional data in order to be clearly constrained.

Human artificial chromosomes (HAC) are a valuable tool in the analysis of complex chromatin structures such as the human centromere because of their small size and relative simplicity compared with normal human chromosomes. This report includes a comprehensive study of the centromere and chromatin composition of HAC, expressing human genes, generated in human cells and transferred to murine cells. The analysis involved chromatin immuno-precipitation and immuno-FISH on metaphase chromosomes and chromatin fibres. In both the cell types, the HAC consisted of alphoid and non-alphoid DNA and were mainly euchromatic in composition, although a pericentromeric heterochromatic region was present on all the HAC. Fibre-FISH and chromatin immuno-precipitation data indicated that the position of the centromere differed between HAC in human cells and in murine cells. Our work highlights the importance and utilisation of HAC for understanding the epigenetic aspects of chromosome biology.

The structural setting of the northern Ionian Sea is the result of the collision between the Calabrian Accretionary Wedge (CAW) and the adjacent foreland, i.e. the Apulian Carbonate Platform. The CAW represents a sector of the Apennine accretionary system extending in the Ionian Sea, bounded to the west by the Malta Escarpment and to the east by the Apulia Escarpment. This work presents the results of the interpretation of new seismic and bathymetric data acquired on the north-eastern edge of the CAW, in the N-Ionian Sea. The data interpretation has identified four main structural domains from NE to SW: 1. The Apulian carbonate Platform consisting of foreland shelf and transitional Mesozoic-Cenozoic carbonate deposits; 2. A narrow foredeep basin, filled by a very thick Plio-Quaternary succession; 3. A deformed domain, at the front of the CAW, incorporating thrusted foredeep sequences and a carbonate block of the Apulian Platform (Transpressed Apulian Block, TAB); 4. A highly deformed pre-Pliocene accretionary wedge. A mid-Pliocene unconformity interpreted on both the CAW and Apulian Foreland suggests that a regional tectonic event occurred at that time, related to the evolution of Calabrian Arc, moving on the subducting oceanic Ionian slab before the collision. This event would correspond to a main tilting and faulting phase of the Apulian Foreland during the diachronous oblique collision with the CAW. The collision and the presence of a remnant of Ionian foreland at the southern front of the accretionary prism, caused a gradual transition to a transpressional tectonics which produced the uplift of the TAB. The TAB would be the south-east continuation of the Amendolara ridge transpressed structure, which forms the offshore extension of the Pollino range. This transpressed shear zone involving the Apulian Foreland developed above the transition between the Adriatic continental crust and the subducting Ionian oceanic crust.

Biomphalaria snails are instrumental in transmission of the human blood fluke Schistosoma mansoni. With the World Health Organization's goal to eliminate schistosomiasis as a global health problem by 2025, there is now renewed emphasis on snail control. Here, we characterize the genome of Biomphalaria glabrata, a lophotrochozoan protostome, and provide timely and important information on snail biology. We describe aspects of phero-perception, stress responses, immune function and regulation of gene expression that support the persistence of B. glabrata in the field and may define this species as a suitable snail host for S. mansoni. We identify several potential targets for developing novel control measures aimed at reducing snail-mediated transmission of schistosomiasis., Biomphalaria glabrata is a fresh water snail that acts as a host for trematode Schistosoma mansoni that causes intestinal infection in human. This work describes the genome and transcriptome analyses from 12 different tissues of B glabrata, and identify genes for snail behavior and evolution.

We show how a bird’s-eye view of genomic structure can be obtained at ∼1-kb resolution from long (∼2 Mb) DNA molecules extracted from whole chromosomes in a nanofluidic laboratory-on-a-chip. We use an improved single-molecule denaturation mapping approach to detect repetitive elements and known as well as unique structural variation. Following its mapping, a molecule of interest was rescued from the chip; amplified and localized to a chromosome by FISH; and interrogated down to 1-bp resolution with a commercial sequencer, thereby reconciling haplotype-phased chromosome substructure with sequence.

The innate and adaptive immune systems of vertebrates possess complementary, but intertwined functions within immune responses. Receptors of the mammalian innate immune system play an essential role in the detection of infected or transformed cells and are vital for the initiation and regulation of a full adaptive immune response. The genes for several of these receptors are clustered within the leukocyte receptor complex (LRC). The purpose of this study was to carry out a detailed analysis of the chicken (Gallus gallus domesticus) LRC. Bacterial artificial chromosomes containing genes related to mammalian leukocyte immunoglobulin-like receptors were identified in a chicken genomic library and shown to map to a single microchromosome. Sequencing revealed 103 chicken immunoglobulin-like receptor (CHIR) loci (22 inhibitory, 25 activating, 15 bifunctional, and 41 pseudogenes). A very complex splicing pattern was found using transcript analyses and seven hypervariable regions were detected in the external CHIR domains. Phylogenetic and genomic analysis showed that CHIR genes evolved mainly by block duplications from an ancestral inhibitory receptor locus, with transformation into activating receptors occurring more than once. Evolutionary selection pressure has led not only to an exceptional expansion of the CHIR cluster but also to a dramatic diversification of CHIR loci and haplotypes. This indicates that CHIRs have the potential to complement the adaptive immune system in fighting pathogens., Synopsis The immune system developed to cope with a diverse array of pathogens, including infectious organisms. The detection of these pathogens by cells of the immune system is mediated by a large set of specific receptor proteins. Here the authors seek to understand how a particular subset of cell surface receptors of the domestic chicken, the chicken Ig-like receptors (CHIR), has evolved. They demonstrate that at least 103 such receptor loci are clustered on a single microchromosome and provide the first detailed analysis of this region. The sequences of the CHIR genes suggest the presence of inhibitory, activating, and bifunctional receptors, as well as numerous incomplete loci (pseudogenes) that appear to have evolved by duplications of an ancestral inhibitory receptor gene. Multiple regions of very high sequence variability were also identified within CHIR loci which, together with considerable expansion of the number of these genes, suggest that CHIR polypeptides are involved in critical functions in the immune system of the chicken.

This chapter concentrates on the use of fluorescence in situ hybridization (FISH) for genomic investigations in the laboratory rat (Rattus norvegicus). The selection of protocols included in the chapter has been inspired by a comprehensive range of previously published molecular cytogenetic studies on this model organism, reporting examples of how FISH can be applied for diverse investigative purposes, varying from comparative gene mapping to studies of chromosome structure and genome evolution, to characterization of chromosomes aberrations as well as transgenic insertions. The protocols, which include techniques for the preparation of mitotic chromosomes and DNA fibers from short-term cell cultures, have been gathered through the years and repeatedly tested in our laboratory, and all together aim at providing sufficient experimental versatility to cover a broad range of cytogenetic and cytogenomic applications.

Summary Pore-forming proteins insert from solution into membranes to create lesions, undergoing a structural rearrangement often accompanied by oligomerization. Lysenin, a pore-forming toxin from the earthworm Eisenia fetida, specifically interacts with sphingomyelin (SM) and may confer innate immunity against parasites by attacking their membranes to form pores. SM has important roles in cell membranes and lysenin is a popular SM-labeling reagent. The structure of lysenin suggests common ancestry with other pore-forming proteins from a diverse set of eukaryotes and prokaryotes. The complex with SM shows the mode of its recognition by a protein in which both the phosphocholine headgroup and one acyl tail are specifically bound. Lipid interaction studies and assays using viable target cells confirm the functional reliance of lysenin on this form of SM recognition., Highlights ► We report the structure of lysenin, a sphingomyelin (SM)-binding pore-forming protein ► Lysenin binds SM's headgroup and tail; the tail occupies the edge of a β sheet ► Lipid binding and live cell studies confirm activity dependence on SM recognition ► Lysenin is related to pore-forming proteins from disparate kingdoms, Lysenin is related to pore-forming proteins found in bacteria and fungi. De Colibus et al. describe structures of lysenin in different Apo conformations and bound to the lipid sphingomyelin. They map its evolutionary relationships and validate structural insights using affinity assays and live cell imaging.

International audience; The ICF syndrome is a rare autosomal recessive disorder, the most common symptoms of which are immunodeficiency, facial anomalies and cytogenetic defects involving decondensation and instability of chromosome 1, 9 and 16 centromeric regions. ICF is also characterised by significant hypomethylation of the classical satellite DNA, the major constituent of the juxtacentromeric heterochromatin. Here we report the first attempt at analysing some of the defining genetic and epigenetic changes of this syndrome from a nuclear architecture perspective. In particular, we have compared in ICF ( Type 1 and Type 2) and controls the large-scale organisation of chromosome 1 and 16 juxtacentromeric heterochromatic regions, their intra-nuclear positioning, and co-localisation with five specific genes (BTG2, CNN3, ID3, RGS1, F13A1), on which we have concurrently conducted expression and methylation analysis. Our investigations, carried out by a combination of molecular and cytological techniques, demonstrate the existence of specific and quantifiable differences in the genomic and nuclear organisation of the juxtacentromeric heterochromatin in ICF. DNA hypomethylation, previously reported to correlate with the decondensation of centromeric regions in metaphase described in these patients, appears also to correlate with the heterochromatin spatial configuration in interphase. Finally, our findings on the relative positioning of hypomethylated satellite sequences and abnormally expressed genes suggest a connection between disruption of long-range geneheterochromatin associations and some of the changes in gene expression in ICF. Beyond its relevance to the ICF syndrome, by addressing fundamental principles of chromosome functional organisation within the cell nucleus, this work aims to contribute to the current debate on the epigenetic impact of nuclear architecture in development and disease.

Chromosome band 10q24 is rich in genes involved in development, tumorigenesis, neurological disorders, hormone metabolism, and environmentally induced disease susceptibility. We have constructed an STS-based integrated physical and genetic map of 10q24 derived from the CEPH-Généthon mega-YAC contig data for this region. This map consists of 42 fluorescence in situ hybridization-mapped overlapping CEPH mega-YACs spanning approximately 15 Mb to which 49 STS markers have been assigned, including 24 Généthon CA repeat genetic markers, 10 known gene loci from the 10q24 region (IFI56, IDE, PDE6C, RBP4, CYP2C, CD39, DNTT, GOT1, WNT8B, and PAX2) and 11 additional expressed sequences of unknown function.

X-linked recessive hypoparathyroidism, due to parathyroid agenesis, has been mapped to a 906-kb region on Xq27 that contains 3 genes (ATP11C, U7snRNA, and SOX3), and analyses have not revealed mutations. We therefore characterized this region by combined analysis of single nucleotide polymorphisms and sequence-tagged sites. This identified a 23- to 25-kb deletion, which did not contain genes. However, DNA fiber-FISH and pulsed-field gel electrophoresis revealed an approximately 340-kb insertion that replaced the deleted fragment. Use of flow-sorted X chromosome-specific libraries and DNA sequence analyses revealed that the telomeric and centromeric breakpoints on X were, respectively, approximately 67 kb downstream of SOX3 and within a repetitive sequence. Use of a monochromosomal somatic cell hybrid panel and metaphase-FISH mapping demonstrated that the insertion originated from 2p25 and contained a segment of the SNTG2 gene that lacked an open reading frame. However, the deletion-insertion [del(X)(q27.1) inv ins (X;2)(q27.1;p25.3)], which represents a novel abnormality causing hypoparathyroidism, could result in a position effect on SOX3 expression. Indeed, SOX3 expression was demonstrated, by in situ hybridization, in the developing parathyroid tissue of mouse embryos between 10.5 and 15.5 days post coitum. Thus, our results indicate a likely new role for SOX3 in the embryonic development of the parathyroid glands.

Simple DNA repeats and sex chromosome differentiation in Asellus aquaticus (Crustacea, Isopoda)

Background Physical mapping of transgenic insertions by Fluorescence in situ Hybridization (FISH) is a reliable and cost-effective technique. Chromosomal assignment is commonly achieved either by concurrent G-banding or by a multi-color FISH approach consisting of iteratively co-hybridizing the transgenic sequence of interest with one or more chromosome-specific probes at a time, until the location of the transgenic insertion is identified. Results Here we report a technical development for fast chromosomal assignment of transgenic insertions at the single cell level in mouse and rat models. This comprises a simplified 'single denaturation mixed hybridization' procedure that combines multi-color karyotyping by Multiplex FISH (M-FISH), for simultaneous and unambiguous identification of all chromosomes at once, and the use of a Quantum Dot (QD) conjugate for the transgene detection. Conclusions Although the exploitation of the unique optical properties of QD nanocrystals, such as photo-stability and brightness, to improve FISH performance generally has been previously investigated, to our knowledge this is the first report of a purpose-designed molecular cytogenetic protocol in which the combined use of QDs and standard organic fluorophores is specifically tailored to assist gene transfer technology.

Low-copy repeats (LCRs) constitute 5% of the human genome. LCRs act as substrates for non-allelic homologous recombination (NAHR) leading to genomic structural variation. The aim of this study was to assess the potential of Fiber-FISH for LCRs direct visualization to support investigations of genome architecture within these challenging genomic regions. We describe a set of Fiber-FISH experiments designed for the study of the LCR22-2. This LCR is involved in recurrent reorganizations causing different genomic disorders. Four fosmid clones covering the entire length of the LCR22-2 and two single-copy BAC-clones, delimiting the LCR22-2 proximally and distally, were selected. The probes were hybridized in different multiple color combinations on DNA fibers from two karyotypically normal cell lines. We were able to identify three distinct structural haplotypes characterized by differences in copy-number and arrangement of the LCR22-2 genes and pseudogenes. Our results show that Multicolor Fiber-FISH is a viable methodological approach for the analysis of genome organization within complex LCR regions., Highlights ► We describe a Fiber-FISH experiment to analyze the LCR22-2 genomic organization. ► Three different haplotypes were identified in two cell lines. ► Fiber-FISH enables the analysis of the genome architecture of complex LCRs.

A detailed reconstruction of the morphology and recent tectonic activity of the Northern Ionian basin is provided on the basis of newly acquired high-resolution swath bathymetric and single and multichannel seismic profiles. The tectonic domains in this area are the Calabrian accretionary wedge and the Apulian foreland. The contact between the two domains, oriented NW–SE, morphologically coincides with a sea-bottom erosional channel (Taranto canyon) characterized on both sides by mass movements induced by slope instability. Along the accretionary wedge/Apulian foreland boundary three different morphological sectors have been recognized whose main characters are represented by a southward stepped increase of erosional canyon activity resulting in a pronounced slope acclivity and a superficial sediment instability. By means of seismic data we correlate the morphology of the sea-bottom to different contractional and flexural processes of the accretionary wedge/foreland system. The interaction between the different rheological domains in the subduction/collision processes could have also induced horizontal stress represented by areas of tectonic release. In an area like this, where very few detailed surveys have been carried out, this study represents the first attempt in correlating the recent tectonic activity to the morphological features and in locating possible slope instability that has to be evaluated for the positioning of offshore infrastructures.

OBJECTIVE Digenic causes of human disease are rarely reported. Insulin via its receptor, which is encoded by INSR, plays a key role in both metabolic and growth signaling pathways. Heterozygous INSR mutations are the most common cause of monogenic insulin resistance. However, growth retardation is only reported with homozygous or compound heterozygous mutations. We describe a novel translocation [t(7,19)(p15.2;p13.2)] cosegregating with insulin resistance and pre- and postnatal growth deficiency. Chromosome translocations present a unique opportunity to identify modifying loci; therefore, our objective was to determine the mutational mechanism resulting in this complex phenotype. RESEARCH DESIGN AND METHODS Breakpoint mapping was performed by fluorescence in situ hybridization (FISH) on patient chromosomes. Sequencing and gene expression studies of disrupted and adjacent genes were performed on patient-derived tissues. RESULTS Affected individuals had increased insulin, C-peptide, insulin–to–C-peptide ratio, and adiponectin levels consistent with an insulin receptoropathy. FISH mapping established that the translocation breakpoints disrupt INSR on chromosome 19p15.2 and CHN2 on chromosome 7p13.2. Sequencing demonstrated INSR haploinsufficiency accounting for elevated insulin levels and dysglycemia. CHN2 encoding β-2 chimerin was shown to be expressed in insulin-sensitive tissues, and its disruption was shown to result in decreased gene expression in patient-derived adipose tissue. CONCLUSIONS We present a likely digenic cause of insulin resistance and growth deficiency resulting from the combined heterozygous disruption of INSR and CHN2, implicating CHN2 for the first time as a key element of proximal insulin signaling in vivo.

We tested the reroductive compatibility existing between four geographically different populations of Proaselis coxalis (Crustacea Isopoda) collected in the river Sarno (near Naples) and on three Sicilian sites: San Domenico; Villa Grazia di Carini and Enna. By means of laboratory hybridization experiments (“no-choice” method) we investigated and characterized the reproductive isolation established between these populations under allopatric conditions. The populations San Domenico and Enna are reproductively isolated from Villa Grazia di Carini and Sarno, so that they may be considered as belonging to different species. The mechanism of reproductive isolation are of postzygotic nature only: embronic death, hbrid inviability and sterility of the surviving hybrids. These results agree with tie theory of aioatric speciation, according to which in the first phase of speciation only post-zygotic mechanisms occur as an accidental product of genetic divergence.

The analyses of about 800 km of Chirp sub-bottom profilers and 600 km2 of Multibeam data acquired during the 2005 and 2007 surveys of the R/VOGS Explora, and their correlation with one new, and several public, multichannel seismic profiles, allow us to propose a relation between the distribution of gas seepages, fracture systems and deep salt features present in the Central Adriatic Sea. Gas seepage is evident from pockmarks on the seabed and in the shallow sub-bottom, where acoustic chimneys and bright spots have been highlighted and analyzed. The Mid-Adriatic Depression (MAD) is a distinct morphological feature in the Central Adriatic Sea elongated in a NE-SW direction. The area is affected by salt doming of Triassic evaporites which cause the two main alignments of the Mid-Adriatic Ridge as far as the Palagruza High and the Jabuka Ridge. These salt tectonics have existed since, at least, Paleogene times and are still active: they characterize sectors with less resistance to deformation produced by successive regional compressive regimes that have affected the area differently during the different geodynamic phases. Gas- seep features are distributed preferentially above and along the fracture systems produced above and around the salt mounds.

The understanding of the evolution of the Antarctic Ice Sheet is crucial for the comprehension of the history of past global climate. The debate regarding the age of the transition to modern “dry- and cold-based” ice sheet after the Neogene polythermal conditions has taken place over more than 20 years. An evident change in the geometry of the depositional systems of the Prydz Bay continental margin demarked the initiation of the Prydz Channel Fan and has been inferred to correspond to this transition. The improvement in the age placement of this change contributes to unravel the last stages of the Antarctic glacial history. We predicted the spatial distribution of P-wave velocity data along both dip- and strike-oriented seismic profiles that intersect 3 Ocean Drilling Program (ODP) sites on the Prydz Bay continental shelf. We used this information to assist the correlation of the existing litho- and bio-stratigraphic information among the drilling sites and to produce an accurate geometric reconstruction of the Neogene shelf units through depth-migration of the seismic data. The revised stratigraphy that we obtained suggests an early late to late early Pliocene age for the seismic reflector at the base of the Prydz Channel Fan. This age, younger than previously proposed, is consistent with the age inferred for similar geometric changes identified in different Antarctic margins.

Petrophysical properties (wet bulk density, porosity, P-wave velocity) are used to predict biogenic silica contents along a seismic reflection profile that ties two well sites, 1095 and 1096, drilled by Ocean Drilling Program (ODP) Leg 178 on sediment drifts on the Pacific continental margin of the Antarctic Peninsula. The biogenic silica contents along the seismic reflection profile were estimated on the basis of three hypotheses about petrophysical properties distributions in the two boreholes and statistical relationships between biogenic silica and other petrophysical properties, which were established on various sediment layers within the boreholes. Our study demonstrates the possibility to reliably predict the distribution of biogenic silica in the sub-seabed sediments if seismic data processed with amplitude preservation are used and statistical relations are considered. We conclude that the statistical extrapolation of biogenic silica content along seismic reflection profiles tied to borehole data is an efficient tool to quantify the amounts of silica undergoing crystalline transformation, which may have strong implications for submarine slope destabilisation.

In order to investigate the replication timing properties of PCDH11X and PCDH11Y, a pair of protocadherin genes located in the hominid-specific non-pseudoautosomal homologous region Xq21.3/Yp11.2, we conducted a FISH-based comparative study in different human and non-human primate (Gorilla gorilla) cell types. The replication profiles of three genes from different regions of chromosome X (ZFX, XIST and ATRX) were used as terms of reference. Particular emphasis was given to the evaluation of allelic replication asynchrony in relation to the inactivation status of each gene. The human cell types analysed include neuronal cells and ICF syndrome cells, considered to be a model system for the study of X inactivation. PCDH11 appeared to be generally characterized by replication asynchrony in both male and female cells, and no significant differences were observed between human and gorilla, in which this gene lacks X-Y homologous status. However, in differentiated human neuroblastoma and cerebral cortical cells PCDH11X replication profile showed a significant shift towards allelic synchrony. Our data are relevant to the complex relationship between X-inactivation, as a chromosome-wide phenomenon, and asynchrony of replication and expression status of single genes on chromosome X.

As the definition of contourites has widened to embrace a large spectrum of sediments in so-called mixed systems, the distinction between contourites and turbidites has become at times vague. The case history of sediment Drift 7 off the Antarctic Peninsula is analysed in this paper in the light of newly acquired swath bathymetry data. The co-existence of various sedimentary processes is reflected in a complex morphology: erosional gullies produced by debris flows on the upper part of the continental slope; deeply incised channels at the slope base; main trunk-type inter-drift turbidity channels separating the drifts; slide scars; undulating depositional bedforms interpreted as bottom-current sediment waves; fluid escape structures perhaps associated with deep-water coral bioherms. The data suggest that Drift 7 is a genuine sediment drift in which bottom currents pirate the sediment of the turbidity currents. Finally, we propose that the control on location and elongation of the drift is inherited from an older margin structure. The relationships between bottom current and deposition are investigated through a comparison with the SE Greenland continental margin, an analogous counterpart in the northern

International audience; Investigation of sequence variation in common inbred mouse strains has revealed a segmented pattern in which regions of high and low variant density are intermixed. Furthermore, it has been suggested that allelic strain distribution patterns also occur in well defined blocks and consequently could be used to map quantitative trait loci (QTL) in comparisons between inbred strains. We report a detailed analysis of polymorphism distribution in multiple inbred mouse strains over a 4.8-megabase region containing a QTL influencing anxiety. Our analysis indicates that it is only partly true that the genomes of inbred strains exist as a patchwork of segments of sequence identity and difference. We show that the definition of haplotype blocks is not robust and that methods for QTL mapping may fail if they assume a simple block-like structure.

Analysis of physical properties measured on cores and on discrete samples collected by the Ocean Drilling Programme (ODP) Leg 178 on the Pacific margin of the Antarctic Peninsula reveals anomalous down‐hole curves of porosity, density, water content, and P‐wave velocity. These indicate an overall trend of increasing porosity with depth and suggest that the drifts are mostly undercompacted. In one of the two boreholes analysed, a sharp decrease in porosity, matching increasing bulk sediment density and increasing compressional velocity occurs towards the base of the hole, which corresponds to a bottom‐simulating reflector in the seismic section. Analysis of seismic reflection, down‐hole logging, geotechnical and mineralogical data from two drilling sites indicates that the observed anomalous consolidation trends are a consequence of the presence of biogenic silica (diatom and radiolarian skeletons) even with a small to moderate amount. Above the bottom‐simulating reflector, intergranular contacts among whole or broken siliceous microfossils prevent normal sediment consolidation. Diagenetic alteration of biogenic opal‐A to opal‐CT causes a dramatic reduction of intra‐ and interskeletal porosity allowing sediments to consolidate at depth. This results in overpressuring and a decrease in the effective stress. Excess fluids are expelled towards the sediment surface through near vertical, small throw normal faults extending from the diagenetic front to the seafloor and affecting the stability of the submarine slope in the form of gravitational creep along a weakened surface. This work shows how physical properties of shallow fine‐grained marine sediments can be analysed as basin‐wide indicators of biogenic silica abundance. The diagenetic alteration of siliceous microfossils is a possible cause of slope instability along world continental margins where bottom‐simulating reflectors related to silica diagenesis are present at a regional scale.

The FP7-funded SiteChar project examined the entire CO 2 geological storage site characterisation process, from the initial feasibility studies through to the final stage of application for a CO 2 storage permit based on criteria defined by the relevant European legislation. The SiteChar workflow for CO 2 geological storage site characterisation provides a description of all elements of a site characterisation study, as well as guidance to streamline the site characterisation process and make sure that the output covers the aspects mentioned in the European Community (EC) Storage Directive. Five potential European storage sites, representative of prospective geological contexts, were considered as test sites for the research work: a North Sea multi-store site (hydrocarbon field and aquifer) offshore Scotland; an onshore aquifer in Denmark; an onshore gas field in Poland; an aquifer offshore in Norway; and an aquifer in the Southern Adriatic Sea. This portfolio combines complementary sites that allowed to encompass the different steps of the characterisation workflow. A key innovation was the development of internal ‘dry-run’ permit applications at the Danish and Scottish sites and their review by relevant regulatory authorities. This process helped to refine the site characterisation workflow, and aimed to identify remaining gaps in site-specific characterisation, needed to secure storage permits under the EC Storage Directive as implemented in ‘host’ Member States. SiteChar considered the important aspect of the public awareness and public opinions of these new technologies, in parallel to technical issues, on the onshore Polish and offshore Scottish sites. A new format to assist public opinion-forming processes was tested involving a small sample of local communities. Generic as well as site-specific information was made available to the general and local public via the internet and at information meetings. These exercises provide insight in the way implementation of CCS project plans may be perceived by local stakeholders, and inform approaches to develop effective local communication and participation strategies. Key lessons from the research conducted in SiteChar were developed as technical recommendations for storage site characterisation and best practice guidance for storage permitting from the perspective of both applicant and regulator. A best practice guide for policymakers and regulators at Member States and European levels, potential storage site developers and operators has been presented. © F. Delprat-Jannaud et al.

Carbon Capture and Storage (CCS) should be a key technology in order to achieve a decline in the CO2 emissions intensity of the power sector and other intensive industry, but this potential deployment could be restricted by cost issues as the International Energy Agency (IEA) in their last projections (World Energy Outlook 2013) has considered only around 1% of global fossil fuel-fired power plants could be equipped with CCS by 2035. The SiteChar project funded by 7th Framework Programme of European Commission gives the opportunity to evaluate the most influential parameters of techno-economic evaluations of four feasible European projects for CO2 geological storage located onshore and offshore and related to aquifer storage or oil and gas reservoirs, at different stages of characterization. Four potential CO2 storage sites have been assessed in terms of storage costs per tonne of CO2 permanently stored (equivalent cost based). They are located offshore UK, onshore Denmark, offshore Norway and offshore Italy. The four SiteChar techno-economic evaluations confirm it is not possible to derive any meaningful average cost for a CO2 storage site. The results demonstrate that the structure of costs for a project is heterogeneous and the storage cost is consequently site dependent. The strategy of the site development is fundamental, the technical choices such as the timing, rate and duration of injection are also important. The way monitoring is managed, using observation wells and logging has a strong impact on the estimated monitoring costs. Options to lower monitoring costs, such as permanent surveys, exist and should be further investigated. Table 1 below summarizes the cost range in Euro per tonne (Discount Rate (DR) at 8%) for the different sites, which illustrates the various orders of magnitude due to the specificities of each site. These figures have how to be considered with care. In particular the Italian and Norwegian sites present very specific features that explain the high estimated costs. For the Italian site, the short duration of CO2 injection associated with a low injection rate makes the CO2 project comparable to a demo project. The Norwegian site is an offshore site located in a virgin area with high infrastructure costs and a combination of injection duration and injection rate that makes the derived costs very sensitive to the discount rate. Table 1. Summary of the cost range in Euro per tonne (discount rate at 8%) €/t CO2 Equivalent storage cost at 8% DR Injectivity (Mt CO2/year) Injection duration (year) Base case Base case Base case UK 11.4 5 20 Denmark 3.2 1.5 40 Norway 26.6 1 40 Italy 29 1 10 The results for both UK and Danish sites confirm therefore the value range calculated by the European Technology Platform for Zero Emission Fossil Fuel Power Plants (ZEP). The main uncertainties in the costs are linked both to the choice of economic parameters (e.g. injected quantities, contingencies) and to the technical choice of operations. This has been studied by sensitivity analyses: for example, if an injection rate is halved and the injection duration is doubled, the Equivalent Storage Cost (ESC) increases by 23% (UK case at 8% DR). Introducing a water production well and water treatment facilities also increases the ESC by 23%, at least on an onshore site. Techno-economic assessments were basically carried out using an 8% discount rate. For projects of long lifetime such a rate severely discounts the late cash flow, especially after 40 years, so that a discount rate of around 4% more in logic of public investment. Compared to other studies, it has to be noted that the scope of the SiteChar analysis does not consider compression and pumping cost, nor transportation cost. This simplifies the techno-economic evaluation but it may not adequately reflect the specific conditions of the individual developments and, hence, distort the comparison between different cases. Lastly, techno-economic evaluation poses questions to policy makers about the real lifetime of a CO2 storage project: what should be the abandon phase and the associated cost and what is the real value of the liability transfer after 20 years of storage? This issue is still an open question, which has been addressed in SiteChar assuming the same approach as ZEP (2011). To counterbalance these CO2 storage costs, policy makers have to set up incentives, either through ETS (Emission Trading System) credits, tax credits or public funding. To improve the commerciality of CCS, Enhanced Oil Recovery (EOR) should be taken into account in the regulation of CCS, as it is one of the rare sources for revenue from a commodity with a real market value. CO2 storage in a saline aquifer close to oil and gas fields could also be considered as a source for CO2 EOR.