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Several decades have passed since the generation of the first embryonic stem cell (ESC) lines both in mice and in humans. Since then, stem cell biologists have tried to understand their potential biological and clinical uses for their implementation in regenerative medicine. The hematopoietic field was a pioneer in establishing the potential use for the development of blood cell products and clinical applications; however, early expectations have been truncated by the difficulty in generating bonafide hematopoietic stem cells (HSCs). Despite some progress in understanding the origin of HSCs during embryonic development, the reproduction of this process in vitro is still not possible, but the knowledge acquired in the embryo is slowly being implemented for mouse and human pluripotent stem cells (PSCs). In contrast, ESC-derived hematopoietic cells may recapitulate some leukemic transformation processes when exposed to oncogenic drivers. This would be especially useful to model prenatal leukemia development or other leukemia-predisposing syndromes, which are difficult to study. In this review, we will review the state of the art of the use of PSCs as a model for hematopoietic and leukemia development. This work was funded by grants from PID2019-104695RB-I00 and PDC2021-120817-I00from AgenciaEstatal de Investigación (AEI) to AB, ERA PerMed GATA2-HuMo Funding Mechanism (Spain: Acció instrumental de SLT011/18/00006 and SLT011/18/00007 of the Department of Health of the Government of Catalonia to AG and A, the Spanish Ministry of Economy, Industry, and Competitiveness (MINECO PID2020-15591RB-100), La Marató de TV3 (202001-32) and CERCA Programme/Generalitat de Catalunya for institutional support to AG. LGP is a recipient of 2021 FI_B2 00188, AGAUR, Generalitat de Cataluña.

This Science Project (SP) contributes to the estimation of the impacts of the invasive species on the European Biodiversity and Ecosystems. This topic is important for European Green Deal and the new European Biodiversity Strategy. The SP is also linked with the socio-economic issues because of the NIS implications to the local ecosystems and their services, and their societal goods and services. Since many of the above impacts may be of local scale, they may alter common practices in circular economies. The SP is implemented by: (a) Combining different sources of data and information; (b) Using a dual workflow to analyse the data; (c) Integrating its resources with core EOSC services and potentially horizontal services available; (d) Engaging the relevant scientific communities. The users will be able to: (a) Analyse distribution patterns of invasive species from different sources of data; (b) Compare the above patterns; (c) Provide managerial suggestions to relevant authorities; (d) Build on the existing infrastructure to address more complex questions (e.g. future scenarios).

Human pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), have enabled the modeling of human development and helped to illuminate mechanisms of monogenic disease, complex disease, and cancer. Lately, the striking ability of PSCs to self-organize has engendered three-dimension (3D) models of human development. The 3D embryonic cell models partially reconstruct the complex architecture of mammalian early embryonic structures and therefore hold great potential for stem cell and developmental studies. In this Research Topic, Gordeeva et al. described a 3D embryoid body differentiation model and compared the spatiotemporal growth and patterning dynamics of embryoid bodies formed from different stem cell origins and culture conditions. Min et al. profiled the proteome and the protein phosphorylation of blastoids-blastocyst-like 3D structures derived from extended pluripotent stem cells (EPSC).

The importance of predisposition to leukaemia in clinical practice is being increasingly recognized. This is emphasized by the establishment of a novel WHO disease category in 2016 called "myeloid neoplasms with germline predisposition". A major syndrome within this group is GATA2 deficiency, a heterogeneous immunodeficiency syndrome with a very high lifetime risk to develop myelodysplastic syndrome (MDS) and acute myeloid leukaemia (AML). GATA2 deficiency has been identified as the most common hereditary cause of MDS in adolescents with monosomy 7. Allogenic haematopoietic stem cell transplantation is the only curative option; however, chances of survival decrease with progression of immunodeficiency and MDS evolution. Penetrance and expressivity within families carrying GATA2 mutations is often variable, suggesting that co-operating extrinsic events are required to trigger the disease. Predictive tools are lacking, and intrafamilial heterogeneity is poorly understood; hence there is a clear unmet medical need. On behalf of the ERAPerMed GATA2 HuMo consortium, in this review we describe the genetic, clinical, and biological aspects of familial GATA2-related MDS, highlighting the importance of developing robust disease preclinical models to improve early detection and clinical decision-making of GATA2 carriers. This work was supported by ERA PerMed GATA2-HuMo Funding Mechanism (Spain: Acció instrumental de SLT011/18/00006 of the Department of Health of the Government of Catalonia, to AG and AB; Hungary: ED-18-1-2019-001 grant from the National Research, Development and Innovation Office to CB; and Germany: German Federal Ministry of Education and Research (BMBF) 2018-123/01KU1904 to MWW), ÚNKP-21-2-I-SE-21 and Hungarian National Academy of Scientist Education grant to KL, TKP2021-NVA-15, TKP2021-EGA-24 and EU's Horizon 2020 research and innovation programme under grant agreement no. 739593 and Elixir Hungary to CB, MSCA No 101029927-scGATA2track (H2020-MSCA-IF-2020) to OM-B, the Spanish Ministry of Economy, Industry, and Competitiveness (MINECO PID2020-15591RB-100), La Marató de TV3 (202001-32), FPS Grant 2018 by Fondazione Pisana per la Scienza ONLUS and CERCA Programme/Generalitat de Catalunya for institutional support to AG, Vera and Joseph Dresner Foundation, Evans MDS Foundation, American Lebanese Syrian Associated Charities, and BMBF MyPred 01GM1911A to MWW. Open access funding enabled and organized by ProjektDEAL.

Germline SAMD9 mutations are one of the most common alterations that predispose to pediatric myelodysplastic syndrome (MDS), a clonal disorder characterized by ineffective hematopoiesis, increasing the risk of developing acute myeloid leukemia (AML). Up to date, a disease model to study the role of SAMD9 mutation in MDS is still lacking. Here, we have generated a human induced pluripotent stem cell (hiPSC) line carrying SAMD9mut (p.I1567M), taking advantage of CRISPR/Cas9 system. As a result, the genetic engineered hiPSC line represent a new in vitro disease model to understand the impact of SAMD9 mutation at molecular and cellular level during hematopoiesis.

EMODnet Chemistry is one of the seven thematic portals of EMODnet (European Marine Observation and Data Network), the long-term initiative aiming to ensure that European marine data are findable, accessible, interoperable and re-usable. EMODnet was launched by DG MARE in 2009 as the pillar of the Blue Growth strategy, Marine Knowledge 2020.Eutrophication (e.g. nutrients, oxygen and chlorophyll), contaminants (e.g. hydrocarbons, pesticides, heavy metals, antifoulants) and marine litter (e.g. beach litter, seafloor litter and floating micro litter) are the main categories of quality assured marine data sets and data products made available through the EMODnet Chemistry portal.45 marine research and monitoring institutes and oceanographic data management experts from 30 countries comprise the EMODnet Chemistry network, including National Oceanographic Data Centres (NODC), National Environmental Monitoring Agencies and Marine Research Institutes actively involved in managing, processing and providing access to data sets from European marine waters and global oceans.During 2020 EMODnet Chemistry consolidated fundamental international collaborations and upgraded cooperation actions on the European and global level to share and harmonize data, knowledge and services, following decision-makers’ needs to implement EU directives, such as MSFD, MSPD, INSPIRE directive, and the Agenda 2030 Sustainable Development Goals of the United NationsMain EMODnet Chemistry 2020 transnational cooperation actions are:The MSFD Technical Group on Marine Litter used the EMODnet Chemistry Marine Litter Database to compute the EU beach litter quantitative Baselines and Threshold values. The European Environment Agency confirmed the use of EMODnet Chemistry data for three environmental state indicators relating to eutrophication and contaminants. Mercator Ocean International and EMODnet Chemistry set up the first joint portfolio of products in support of the MSFD implementation. The two partners are also exploring opportunities to support the aquaculture sector. EMODnet -Chemistry and the In Situ Thematic Assembly Centre of the Copernicus Marine Environment Monitoring Service (CMEMS INSTAC) collaborated with ENVRI Marine European Research Infrastructures (Euro-Argo, EMSO, ICOS, Lifewatch and SeaDataNet) to enhance FAIRness of in situ data. Mercator Ocean international, UNDESA, SULITEST NGO and EMODnet Chemistry have been creating an awareness questionnaire to raise awareness on the Goal 14 of the UN Agenda 2030 for Sustainable Development. The EU asked EMODnet Chemistry to share its experience at the G20 workshop on harmonized monitoring and data compilation of marine plastic litter organized by the Ministry of the Environment, Japan. The international Oxygen data portal and Ocean Acidification data portal received contributions from EMODnet Chemistry and CMEMS in situ TAC for their implementation. The National Marine Data and Information Service of China collaborates with EMODnet to strengthen international ocean data through the EMOD-PACE project.

Lung cancer is the leading cause of cancer deaths. Tumor heterogeneity, which hampers development of targeted therapies, was herein deconvoluted via single cell RNA sequencing in aggressive human adenocarcinomas (carrying Kras-mutations) and comparable murine model. We identified a tumor-specific, mutant-KRAS-associated subpopulation which is conserved in both human and murine lung cancer. We previously reported a key role for the oncogene BMI-1 in adenocarcinomas. We therefore investigated the effects of in vivo PTC596 treatment, which affects BMI-1 activity, in our murine model. Post-treatment, MRI analysis showed decreased tumor size, while single cell transcriptomics concomitantly detected near complete ablation of the mutant-KRAS-associated subpopulation, signifying the presence of a pharmacologically targetable, tumor-associated subpopulation. Our findings therefore hold promise for the development of a targeted therapy for KRAS-mutant adenocarcinomas., Maroni, Bassal, Krishnan et al. characterise human non-small cell lung cancer (NSCLC) carrying Kras-mutations by single-cell RNA sequencing. They identify a tumour-specific population that is conserved in mice and responds to the drug, PTC596, which is currently in clinical trials and may offer a potential avenue for treating aggressive NSCLC expressing mutated Kras.

A major challenge in regenerative medicine is the generation of functionally effective target cells to replace or repair damaged tissues. The finding that most somatic cells can be directly converted into cells of another lineage by the expression of specific transcription factors has paved the way to novel applications. Induced neurons (iNs) represent an alternative source of neurons for disease modeling, drug screening, and potentially, for cell replacement therapy. This unit describes methods for the efficient conversion of blood cells into iNs, including protocols to isolate cord blood CD133+ cells, infect them with Sendai virus vectors that express SOX2 and c-MYC, and differentiate the infected cells (PB-MNCs) into mature neurons. A method to reprogram peripheral blood mononuclear cells into iNs is also described. Support protocols describe how to culture rat astrocytes and characterize the electrophysiology of iNs. © 2016 by John Wiley & Sons, Inc.

High quality and integrated information able to show marine litter distribution at a global scale is a crucial goal to tackle the environmental problem. One of the important gaps is the definition of a global monitoring protocol and reporting. Large data infrastructures can provide a sustainable framework to host harmonized and standard litter data that can be used and re-used for any purpose, including assessment. EMODnet Chemistry has collected marine litter data since 2016 and has adopted different strategies for the management of the diverse litter data types, exploiting the advantages deriving from the application of the FAIR principles in marine litter data stewardship. The quality of the released data sets is improved allowing a better consistency within data values collected in different contexts (several countries, different techniques, …).

Summary In vertebrates, GATA2 is a master regulator of hematopoiesis and is expressed throughout embryo development and in adult life. Although the essential role of GATA2 in mouse hematopoiesis is well established, its involvement during early human hematopoietic development is not clear. By combining time-controlled overexpression of GATA2 with genetic knockout experiments, we found that GATA2, at the mesoderm specification stage, promotes the generation of hemogenic endothelial progenitors and their further differentiation to hematopoietic progenitor cells, and negatively regulates cardiac differentiation. Surprisingly, genome-wide transcriptional and chromatin immunoprecipitation analysis showed that GATA2 bound to regulatory regions, and repressed the expression of cardiac development-related genes. Moreover, genes important for hematopoietic differentiation were upregulated by GATA2 in a mostly indirect manner. Collectively, our data reveal a hitherto unrecognized role of GATA2 as a repressor of cardiac fates, and highlight the importance of coordinating the specification and repression of alternative cell fates., Graphical Abstract, Highlights • GATA2 promotes hemogenic emergence during human iPSC differentiation • GATA2 enhances hematopoietic differentiation of human iPSCs • GATA2 acts as a direct repressor of cardiac fates during mesoderm specification, Giorgetti A. and colleagues demonstrate that GATA2 induction in early mesodermal cells leads to the robust generation of hemogenic endothelial and hematopoietic cells from human iPSCs. They show the ability of GATA2 to instruct mesodermal precursors toward a hematopoietic cell fate and concurrently inhibit cardiac fates. This study expands our understanding of the regulatory networks that control early human hematopoiesis.

Marine data are needed for many purposes: for acquiring a better scientific understanding of the marine environment, but also, increasingly, as marine knowledge for decision making as well as developing products and services supporting economic growth. Data must be of sufficient quality to meet the specific users' needs. It must also be accessible in a timely manner. And yet, despite being critical, this timely access to known-quality data proves challenging. Europe's marine data have traditionally been collected by a myriad of entities with the result that much of our data are scattered throughout unconnected databases and repositories. Even when data are available, they are often not compatible, making the sharing of the information and data aggregation particularly challenging. In this paper, we present how the European Marine Observation and Data network (EMODnet) has developed over the last decade to tackle these issues. Today, EMODnet is comprised of more than 150 organizations which gather marine data, metadata, and data products and make them more easily accessible for a wider range of users. EMODnet currently consists of seven sub-portals: bathymetry, geology, physics, chemistry, biology, seabed habitats, and human activities. In addition, Sea-basin Checkpoints have been established to assess the observation capacity in the North Sea, Mediterranean, Atlantic, Baltic, Artic, and Black Sea. The Checkpoints identify whether the observation infrastructure in Europe meets the needs of users by undertaking a number of challenges. To complement this, a Data Ingestion Service has been set up to tackle the problem of the wealth of marine data that remain unavailable, by reaching out to data holders, explaining the benefits of sharing their data and offering a support service to assist them in releasing their data and making them available through EMODnet. The EMODnet Central Portal (www.emodnet.eu) provides a single point of access to these services, which are free to access and use. The strategic vision of EMODnet in the next decade is also presented, together with key focal areas toward a more user-oriented service, including EMODnet for business, internationalization for global users, and stakeholder engagement to connect the diverse communities across the marine knowledge value chain.

We have generated two human induced pluripotent stem cell (iPSC) lines from CD133

Sensor engineering is continuously evolving as devices become cheaper, smaller, more intelligent, and more efficient. Today, oceanographic sensors aim at monitoring marine processes by means of physical, chemical, and biological variables, and use different data formats, units, parameters, resolutions, data quality standards, and protocols. Therefore, integration and interoperability at European level represent a challenge. To cope with this challenge, the Sensor Web Enablement (SWE) standards, developed by the Open Geospatial Consortium (OGC), are a good solution, as they ensure interoperability and long-term archiving of data series with complete information for all devices. The interoperability allows integrating information from different sources or pre-existing architectures, such as those developed in the SANY project ( http://www.opengeospatial.org/ogc/regions/SANY ) or by the US Integrated Ocean Observing System ( https://github.com/ioos ). In this paper, we illustrate the real-time data management system, developed by the Italian National Oceanographic Data Centre (NODC) at the Istituto Nazionale di Oceanografia e di Geofisica Sperimentale, OGS in Trieste – Italy, designed to share data acquired by a large number of heterogeneous observing platforms. This system adopts Observations and Measurements (O&M) and Sensor Model Language (SensorML) as data and metadata formats, as well as the Sensor Observation Service (SOS) released by the 52°North as server and Web client for open data access. The work done shows that the choice to manage real-time data using OGC Sensor Web Enablement (SWE) standards can be considered a valid solution with pros and cons. Nevertheless, in the future, the SWE community will grow, and with it, the number of applications to manage SWE standards to simplify their adoption.

Recent studies in zebrafish and mice have revealed that proinflammatory signaling is a positive regulator of definitive hematopoietic development. Whether proinflammatory signaling also regulates human hematopoietic specification remains unknown. Here, we explored the impact of the proinflammatory cytokines tumor necrosis factor-α (TNFα), interferon-γ (IFNγ), and interleukin-1β (IL1β) on in vitro hematopoietic differentiation using human pluripotent stem cells. Gene expression analysis and enzyme-linked immunosorbent assay revealed the absence of a proinflammatory signature during hematopoietic development of human pluripotent stem cells. Functionally, the emergence of hemogenic endothelial progenitors (CD31+CD34+CD45- or CD34+CD43-CD73-) and hematopoietic cells (CD43+CD45+) was not affected by treatment with increasing doses of TNFα, IFNγ, and IL1β irrespective of the developmental window or the differentiation protocol used (embryoid body or OP9 co-culture based). Similarly, knockdown of endogenous NF-kB signaling had no impact on hematopoietic differentiation of human pluripotent stem cells. This study serves as a demonstration that TNFα, IFNγ, and IL1β signals do not improve hematopoietic differentiation of human pluripotent stem cells using current protocols and suggests that proinflammatory signaling is insufficient to drive definitive hematopoietic specification of human hematopoietic stem cells in vitro. This work was supported by the European Research Council (646903 to PM) and the Spanish Ministry of Economy and Competitiveness (SAF2013-43072R to AG, SAF2013-43065R to PM, and ISCIII/FEDER-PI14/01119 to CB). CB is supported by a Miguel Servet contract (CPII13/00011). AG is supported by the Ramon y Cajal Program (RyC-2013-13221). PM also acknowledges support from Obra Social La Caixa-Fundacio Josep Carreras and Generalitat de Catalunya (SGR330).

Metadata, by definition, is information associated with data, covering where, how, when and by whom the data were acquired. The chance to take part in European projects such as the EU-SeaDataNet (the Pan-European infrastructure for ocean and marine data management) made it necessary to use XML (Extensible Markup Language) as a standard file format for sharing data and metadata. At present, the Italian National Oceanographic Data Centre (OGS/NODC) has all its data and metadata contained in an Oracle relational database, and the metadata is managed using XML formats and schema (XSD; XML Schema Definition), giving common vocabularies for parameters, instruments, ships, etc. in agreement with European project standards. One problem with XML is the dynamic change in metadata schemas (XSD), necessitating development of a system which is flexible and capable of managing the changes. This paper describes a system for managing oceanographic metadata using XML files and the functionalities provided by the Oracle database. To better manage the XML format, we chose to load into the OGS/NODC database the whole XML file, using a dedicated field. The database Oracle gives us the flexibility to manage the XML format locally, within the OGS/NODC information system, using the XML DB (XML DataBase) Oracle features. This, through the use of XMLType, allows the inclusion of the XML into the database. Furthermore, through the use of the XQuery functions it is possible to create a set of views through which information contained in the XML can be viewed more immediately in a relational form. Using the XML and the XQuery functions, it is possible to store, extract and manage different kinds of information that might be exchanged at the European level. Moreover, with a RESTful (Representational State Transfer) Web Service, we have a simple and standard interface for rapidly and easily creating, modifying and deleting records containing XML files inside the database. Finally, through the use of a RESTful Web Service, it is possible to decouple the application from the database, so that through the use of software that manages HTTP URLs, such as the Mikado (SeaDataNet project), the XML file can be inserted, updated and deleted inside the database without the need for a direct connection to it.

Coenzyme Q

Induced pluripotent stem cells (iPSCs) represent an invaluable tool in a chromosomal instability syndrome such as Fanconi anemia (FA), as they can allow to study of the molecular defects underlying this disease. Many other applications, such as its use as a platform to test different methods or compounds, could also be of interest. But the greatest impact of iPSCs may be in bone marrow failure diseases, as iPSCs could represent an unlimited source of autologous cells to apply in advanced treatments such as gene therapy. At the same time, genome editing constitutes the next generation of technology to further develop a safer, personalized, targeted gene therapy. Despite the promising advantages that these two technologies would present in a disease such as FA, the specific characteristics of the disease make both of these processes especially challenging. Efficient and safer FA-hiPSC (human induced pluripotent stem cell) generation methods, robust and reliable differentiation protocols for iPSCs, as well as really efficient delivery methods to perform targeted gene correction should be developed.

Seasonally and spatially averaged vertical profiles of temperature, salinity, dissolved oxygen, nutrients and chlorophyll- a have been computed from in situ observations in different regions of the Mediterranean Sea using the recently released EU/MEDAR/MEDATLAS II and EU/MTPII/MATER databases. The regions have been defined according to known dynamics important in the formation, transformation and spreading of the main water masses that circulate in the upper, intermediate and deep layers. The climatological characteristics of temperature and salinity reflect the water mass structures and the general thermohaline circulation patterns. Spatial and temporal variations on a seasonal basis of nutrients and principal biological parameters are described along with some aspects of the trophic conditions of the Mediterranean Sea. The strongest signal of variability is along the vertical; however, horizontal inhomogeneities are mostly associated with the internal dynamics. The distribution of the biochemical elements, in conjunction with hydrographic measurements of temperature and salinity, may serve as appropriate tracers for the characterisation of the main water masses (Atlantic Water, Levantine Intermediate Water and Bottom Water in the eastern and eastern basins) throughout the Mediterranean Sea. In a first approach, adequate descriptors of water properties have been obtained, useful for the quality control of incoming data in large databases and in setting-up documented procedures to improve future data management practices. Secondly, the biochemical climatological characteristics addressed in this paper are verified to be consistent with the physics of the Mediterranean Sea and are conceived useful to initialise coupled physical–biological models. The full set of spatially averaged vertical profiles can be found and downloaded from the World Wide Web data server established at OGS ( http://doga.ogs.trieste.it/medar/climatologies/ ).

Data from eight CTD casts and two one-year long current time series collected at 8 and 60 m above the seafloor of a sediment drift, off the Pacific Margin of the Antarctic Peninsula are presented, with special emphasis on bottom boundary layer dynamics and processes relevant to sediment settling and re-suspension. The water masses over the drift are characterized, including also a comparison with other measurements available from that region. The south-westward flow along the continental rise exhibits a strong topographic (bathymetric) control in the near-bottom current regime. A consistent mean flow deflection between an upper and lower current regime suggests that only the lower regime falls within a bottom (turbulent) Ekman layer. The bottom current regime is not energetic enough to maintain the coarse sediment fraction in suspension. The absence of evidence for a nepheloid layer justifies the assumption that most sediment was supplied to the margin during glacial periods. Two events, with peak velocities of up to 20 cm s−1, are associated with barotropic eddies shown as negative (cyclonic) mean sea level anomalies detected by ERS/TOPEX satellite altimeters. These energetic bottom current pulses may give way to episodic sediment re-suspensions of the sortable (non-cohesive) part of the sediment, thus exerting a minor role in redistributing fine sediments through the mean flow regime.

Otx1 belongs to the paired class of homeobox genes and plays a pivotal role in brain development. Here, we show that Otx1 is expressed in hematopoietic pluripotent and erythroid progenitor cells. Moreover, bone marrow cells from mice lacking Otx1 exhibit a cell-autonomous impairment of the erythroid compartment. In agreement with these results, molecular analysis revealed decreased levels of erythroid genes that include the SCL and GATA-1 transcription factors. Accordingly, a gain of function of SCL rescues the erythroid deficiency in Otx1 - / - mice. Taken together, our findings indicate a function for Otx1 in the regulation of blood cell production.

An advanced computer and communication technology was used to develop an integrated system and software tools for managing a great diversity of oceanographic data collected in the Mediterranean Sea during 1996–2000. Data were obtained during 108 sea cruises, carried out within the framework of the large-scale international research project MATER (mass transfer and ecosystem response), which was financially supported by the Marine Science and Technology (MAST) Programme of the European Union (EU). Data collection involved the active participation of various research vessels and personnel coming from 58 different laboratories of 13 countries. Data formatting as well as automatic and visual data quality controls were implemented using internationally accepted standards and procedures. Various data inventories and meta-data information, accessible through the World Wide Web (WWW), are made available to the user community. A database was developed, which, along with meta-data and other data relevant to the project information, is made available to the user community in the form of a CD-ROM. The database consists of 5861 vertical profiles and 842 time series of basic physical and biogeochemical parameters collected in the seawater column as well as biogeochemical parameters from the analysis of 70 sediment cores. Furthermore, it includes 67 cruise data files of nonstandard additional biological and atmospheric parameters.

Twelve sedimentary mounds are identified on the upper continental rise of the Pacific Margin of the Antarctic Peninsula. All these mounds are produced by a varying degree of interaction of along-slope bottom water flow with down-slope turbidity currents. These mounds provide a complete range of intermediates between two end members: the sediment drift and the channel levee. Surface sediments on drift 7 suggest that the mechanisms for the supply and transport of sediment include entrainment of material from turbidity currents within ambient bottom currents, and pelagic settling from the sea surface, including biogenic and glacially derived material. The long-lasting activity of these mechanisms is documented by the data provided by four DSDP and ODP drill sites. Bathymetric and seismic data, both at a large, comprehensive scale and at a small, detailed scale, show the geometry of the sedimentary mounds and their relationships with the adjacent turbidity current channel systems. These data allow the determination of some diagnostic criteria to identify the sediment drifts.

The Ocean Biogeographic Information System (OBIS) aims to integrate smaller, isolated datasets into a larger, more comprehensive picture of life in our oceans. Therefore, OBIS provides a gateway to many datasets containing information on where and when marine species have been observed. The datasets within OBIS are contributed by a network of hundreds of institutes, projects and individuals, all with the common goal to gain scientific knowledge and to make these data and knowledge easily available to the public. Until recently, OBIS had solely focused on biogeographic data, in the form of presence of marine species in space and time. Data collected for biological studies however often include more than just presence or abundance. Physical and chemical measurements are often taken concomitantly providing insights into the environmental conditions the species live in. Details on the nature of the sampling methods, equipment used and effort can also be of major importance. Based on requirements from the growing OBIS community for data archiving and scientific applications, OBIS completed the OBIS-ENV-DATA project in 2017 to enhance its data standard by accommodating additional data types (De Pooter et al. 2017). The proposed standard allows for the management of sampling methodology, animal tracking and telemetry data, and environmental measurements such as nutrient concentrations, sediment characteristics and other abiotic parameters measured during sampling. The new OBIS data standard builds on the Darwin Core Archive and on practices adopted by the Global Biodiversity Information Facility (GBIF). It consists of an Event Core in combination with an Occurrence Extension and an enhanced MeasurementOrFact Extension Fig. 1. This new structure enables the linkage of measurements or facts - quantitative or qualitative properties - to both sampling events and species occurrences, and includes additional fields for property standardization. The OBIS standard also embraces the use of the new Darwin Core term parentEventID, enabling a sampling event hierarchy. During the follow-up project "OBIS-Event Data", the format will be further fine-tuned during two workshops with two different communities of practice. The first workshop (April 2018) will focus on animal tagging and tracking data, while the second one (October 2018) will tackle macro- and meiobenthos data. The OBIS-Event Data project will also develop the first data products and applications based on the standard and make these tools part of the core OBIS data system output. We believe that the adoption of this new data standard by the international community will be key to improving the effectiveness of the knowledge base and will enhance integration and management of critical data needed to understand ecological and biological processes in the ocean.

The Ocean Biogeographic Information System (OBIS) is the world's most comprehensive online, open-access database of marine species distributions. OBIS grows with millions of new species observations every year. Contributions come from a network of hundreds of institutions, projects and individuals with common goals: to build a scientific knowledge base that is open to the public for scientific discovery and exploration and to detect trends and changes that inform society as essential elements in conservation management and sustainable development. Until now, OBIS has focused solely on the collection of biogeographic data (the presence of marine species in space and time) and operated with optimized data flows, quality control procedures and data standards specifically targeted to these data. Based on requirements from the growing OBIS community to manage datasets that combine biological, physical and chemical measurements, the OBIS-ENV-DATA pilot project was launched to develop a proposed standard and guidelines to make sure these combined datasets can stay together and are not, as is often the case, split and sent to different repositories. The proposal in this paper allows for the management of sampling methodology, animal tracking and telemetry data, biological measurements (e.g., body length, percent live cover, ...) as well as environmental measurements such as nutrient concentrations, sediment characteristics or other abiotic parameters measured during sampling to characterize the environment from which biogeographic data was collected. The recommended practice builds on the Darwin Core Archive (DwC-A) standard and on practices adopted by the Global Biodiversity Information Facility (GBIF). It consists of a DwC Event Core in combination with a DwC Occurrence Extension and a proposed enhancement to the DwC MeasurementOrFact Extension. This new structure enables the linkage of measurements or facts - quantitative and qualitative properties - to both sampling events and species occurrences, and includes additional fields for property standardization. We also embrace the use of the new parentEventID DwC term, which enables the creation of a sampling event hierarchy. We believe that the adoption of this recommended practice as a new data standard for managing and sharing biological and associated environmental datasets by IODE and the wider international scientific community would be key to improving the effectiveness of the knowledge base, and will enhance integration and management of critical data needed to understand ecological and biological processes in the ocean, and on land. Fil: De Pooter, Daphnis. Flanders Marine Institute; Bélgica Fil: Appeltans, Ward. UNESCO-IOC; Bélgica Fil: Bailly, Nicolas. Hellenic Centre for Marine Research, MedOBIS; Grecia Fil: Bristol, Sky. United States Geological Survey; Estados Unidos Fil: Deneudt, Klaas. Flanders Marine Institute; Bélgica Fil: Eliezer, Menashè. Istituto Nazionale di Oceanografia e di Geofisica Sperimentale; Italia Fil: Fujioka, Ei. University Of Duke. Nicholas School Of Environment. Duke Marine Lab; Estados Unidos Fil: Giorgetti, Alessandra. Istituto Nazionale di Oceanografia e di Geofisica Sperimentale; Italia Fil: Goldstein, Philip. University of Colorado Museum of Natural History, OBIS; Estados Unidos Fil: Lewis, Mirtha Noemi. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; Argentina Fil: Lipizer, Marina. Istituto Nazionale di Oceanografia e di Geofisica Sperimentale; Italia Fil: Mackay, Kevin. National Institute of Water and Atmospheric Research; Nueva Zelanda Fil: Marin, Maria Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico; Argentina Fil: Moncoiffé, Gwenaëlle. British Oceanographic Data Center; Reino Unido Fil: Nikolopoulou, Stamatina. Hellenic Centre for Marine Research, MedOBIS; Grecia Fil: Provoost, Pieter. UNESCO-IOC; Bélgica Fil: Rauch, Shannon. Woods Hole Oceanographic Institution; Estados Unidos Fil: Roubicek, Andres. CSIRO Oceans and Atmosphere; Australia Fil: Torres, Carlos. Universidad Autonoma de Baja California Sur; México Fil: van de Putte, Anton. Royal Belgian Institute for Natural Sciences; Bélgica Fil: Vandepitte, Leen. Flanders Marine Institute; Bélgica Fil: Vanhoorne, Bart. Flanders Marine Institute; Bélgica Fil: Vinci, Mateo. Istituto Nazionale di Oceanografia e di Geofisica Sperimentale; Italia Fil: Wambiji, Nina. Kenya Marine and Fisheries Research Institute; Kenia Fil: Watts, David. CSIRO Oceans and Atmosphere; Australia Fil: Klein Salas, Eduardo. Universidad Simon Bolivar; Venezuela Fil: Hernandez, Francisco. Flanders Marine Institute; Bélgica

An updated climatology, based on a comprehensive data set (1911–2009) of temperature, salinity and dissolved oxygen, has been produced for the whole Adriatic Sea with the variational inverse method using the DIVA (Data-Interpolating Variational Analysis) software. Climatological maps were produced at 26 levels and validated with ordinary cross-validation and with a real vs. synthetic temperature–salinity diagram intercomparison. The concept of climatology–observation misfit (COM) has been introduced as an estimate of the physical variability associated with the climatological structures. In order to verify the temporal stability of the climatology, long-term variability has been investigated in the Middle Adriatic and the South Adriatic pits, regarded as the most suitable records of possible long-term changes. Compared with previous climatologies, this study allows a clear identification of the seasonal dynamic of the southern Adriatic, where a clear oxygen minimum is typically observed in the centre of the South Adriatic Gyre. New and better resolved features emerged from this analysis: (1) below 100 m all properties profoundly differ between the central and the southern Adriatic and seem characterized by different biogeochemical dynamics; (2) the South Adriatic Pit clearly shows the remote effects of the Eastern Mediterranean Transient, while no effect is observed in the Middle Adriatic Pit; (3) the deepest part of the southern Adriatic seems now to be significantly saltier (+0.18 psu since the period 1910–1914, with an increase of +0.018 decade−1 since the late 1940s) and warmer (+0.54 °C since 1910–1914) even though a long-term temperature trend could not be statistically demonstrated; (4) the Middle Adriatic Pit shows a long-term increase in apparent oxygen utilization (+0.77 mL L−1 since 1910–1914, with a constant increase of +0.2 mL L−1 decade−1 after the 1970s).

The finding that certain somatic cells can be directly converted into cells of other lineages by the delivery of specific sets of transcription factors paves the way to novel therapeutic applications. Here we show that human cord blood (CB) CD133 + cells lose their hematopoietic signature and are converted into CB-induced neuronal-like cells (CB-iNCs) by the ectopic expression of the transcription factor Sox2, a process that is further augmented by the combination of Sox2 and c-Myc. Gene-expression analysis, immunophenotyping, and electrophysiological analysis show that CB-iNCs acquire a distinct neuronal phenotype characterized by the expression of multiple neuronal markers. CB-iNCs show the ability to fire action potentials after in vitro maturation as well as after in vivo transplantation into the mouse hippocampus. This system highlights the potential of CB cells and offers an alternative means to the study of cellular plasticity, possibly in the context of drug screening research and of future cell-replacement therapies.

Defined transcription factors can induce epigenetic reprogramming of adult mammalian cells into induced pluripotent stem cells. Although DNA factors are integrated during some reprogramming methods, it is unknown whether the genome remains unchanged at the single nucleotide level. Here we show that 22 human induced pluripotent stem (hiPS) cell lines reprogrammed using five different methods each contained an average of five protein-coding point mutations in the regions sampled (an estimated six protein coding point mutations per exome). The majority of these mutations were non-synonymous, nonsense, or splice variants, and were enriched in genes mutated or having causative effects in cancers. At least half of these reprogramming-associated mutations pre-existed in fibroblast progenitors at low frequencies, while the rest were newly occurring during or after reprogramming. Thus, hiPS cells acquire genetic modifications in addition to epigenetic modifications. Extensive genetic screening should become a standard procedure to ensure hiPS safety before clinical use.

Generation of human induced pluripotent stem cells (hiPSCs) by the expression of specific transcription factors depends on successful epigenetic reprogramming to a pluripotent state. Although hiPSCs and human embryonic stem cells (hESCs) display a similar epigenome, recent reports demonstrated the persistence of specific epigenetic marks from the somatic cell type of origin and aberrant methylation patterns in hiPSCs. However, it remains unknown whether the use of different somatic cell sources, encompassing variable levels of selection pressure during reprogramming, influences the level of epigenetic aberrations in hiPSCs. In this work, we characterized the epigenomic integrity of 17 hiPSC lines derived from six different cell types with varied reprogramming efficiencies. We demonstrate that epigenetic aberrations are a general feature of the hiPSC state and are independent of the somatic cell source. Interestingly, we observe that the reprogramming efficiency of somatic cell lines inversely correlates with the amount of methylation change needed to acquire pluripotency. Additionally, we determine that both shared and line-specific epigenetic aberrations in hiPSCs can directly translate into changes in gene expression in both the pluripotent and differentiated states. Significantly, our analysis of different hiPSC lines from multiple cell types of origin allow us to identify a reprogramming-specific epigenetic signature comprised of nine aberrantly methylated genes that is able to segregate hESC and hiPSC lines regardless of the somatic cell source or differentiation state.

Induced pluripotent stem cells (iPSC ) provide an invaluable resource for regenerative medicine as they allow the generation/nof patient-specific progenitors with potential value for cell therapy. However, in many instances, an off-the-shelf approach is/ndesirable, such as for cell therapy of acute conditions or when the patient’s somatic cells are altered as a consequence of a chronic/ndisease or aging. Cord blood (CB) stem cells appear ideally suited for this purpose as they are young cells expected to carry/nminimal somatic mutations and possess the immunological immaturity of newborn cells; additionally, several hundred thousand/nimmunotyped CB units are readily available through a worldwide network of CB banks. Here we present a detailed protocol for the/nderivation of CB stem cells and how they can be reprogrammed to pluripotency by retroviral transduction with only two factors/n(OCT 4 and SO X2) in 2 weeks and without the need for additional chemical compounds.

We read with great interest the recent report in the American Journal of Cardiology by Hoetzer et al.1 The investigators addressed the question of whether in healthy, middle-aged subjects circulating endothelial progenitor cell (EPC) number and function (capacity to form colonies) are different in women and men. It was a study well conducted by an experienced team.

The majority of clinical studies on endothelial progenitor cells (EPCs) focuses on the role of these cells in cardiovascular diseases and no systematic studies exist regarding their variations in healthy subjects. In order to define the burden of angiogenesis in physiological conditions we assessed the frequency of peripheral blood endothelial colonies (PB-ECs) and their relation with other factors possibly involved in their function such as high-sensitivity C-reactive protein (hs-CRP), endothelial cell-specific mitogen factor (VEGF) and tissue inhibitor of metalloproteinases-1 (TIMP-1) in a highly selected healthy population. A PB sample was obtained from 37/47 healthy subjects (age 40.2+/-15.0yrs; M/F 15/22) without known cardiovascular risk factors. The serum level of hs-CRP, VEGF, TIMP-1, the frequency of PB-ECs by clonogenic assay, and the number of early EPCs and late EPCs by flow cytometry analysis were evaluated. PB-ECs were formed by 40.5% of studied subjects with a mean of 0.40+/-0.82 colonies/10(6) cells. The differences in the frequency of colony formation between genders were not statistically significant. The subjects with PB-ECs were characterized by higher values of hs-CRP, when compared with those not forming colonies, 0.276+/-0.230 vs 0.095+/-0.077 mg/l (p=0.003) respectively, and of VEGF, 328.3+/-162.9 vs 202.68+/-118.53 pg/ml (p=0.02). No significant differences were found in TIMP-1 values. The EPC clonogenic potential seems to be related to hs-CRP and VEGF levels even in healthy population supporting the concept that these mediators are involved in physiological ECs function.

Reconstructions of Mediterranean ocean temperature fields back to 1950 show a proxy relationship between heat content changes in the North Atlantic and the Western Mediterranean Deep Water (WMDW) formed in the Gulf of Lions in winter, because of consistent air-sea heat fluxes over these areas, strongly correlated to the North Atlantic Oscillation (NAO).

Circulating endothelial progenitor cells (EPCs) are believed to contribute to vascular homeostasis; unfortunately, the response of EPCs in physiological conditions remains largely unknown. Herein we report our observations of a 44-year-old healthy subject after a trek in the Himalayas that support high-altitude hypoxia and exercise oxygen demands are strong stimuli for clonogenic endothelial cell activation and activity, as shown by the increase in the number of mature EPCs and in the endothelial colony-forming unit capacity. Both of these effects were completely reverted at sea level, 45 days after the subject's trek. Am. J. Hematol. 79:76-78, 2005. 2005 Wiley-Liss, In

Significant progress has been achieved during the past 10 years in cell transplantation and recent research has focused on the possibility of improving ventricular function after myocardial infarction. Most studies in the field of cardiac tissue repair are performed by direct intramyocardial injection of cells of different origin. Since this approach requires a surgical intervention, in this study we investigated the feasibility of non-invasive administration of bone marrow mononuclear cells (BMMNCs) by assessing the fate of peripherally injected, purified, labeled cells in cryodamaged hearts.Ten donor and ten recipient inbred isogenic adult (4 weeks old) Fisher rats were used as models to mimic autologous transplantation. Myocardial damage was obtained in recipient rats by placing a frozen metal probe on the anterior left ventricular wall for 15 seconds (freeze-thaw injury technique). BMMNCs were purified and labeled with a red fluorescent cell dye. Seven days after the injury about 15-25x10(6) cells were infused through the femoral vein of recipient rats. Seven days after the infusion, the heart, lungs, liver, kidneys, spleen and thymus were harvested to track transplanted cells.Labeled cells were found only in the injured area of the heart and not in the normal tissue, and a limited number of cells were identified in the spleen of all the animals. Most of the labeled cells in the infarcted area were Thy-1(+) and some were CD34(+).Our data suggest that peripherally injected BMMNCs can traffic through the circulation to the site of damage; we hypothesize that tissue injury leads to the priming of a cytokine cascade acting as chemoattractant for the infused cells.

Traditionally, somatic tissue-derived stem cells of mammalian adults have been viewed as pluripotent precursors capable of lifelong maintenance of cellular compartments typical of the tissue in which they reside. However in recent years, in vitro cultures and in vivo transplantation assays have indicated that adult somatic stem cell of various species are capable of adopting multiple fates. Bone marrow cells can give rise to a wide array of phenotypes, including blood, endothelial, bone, cartilage, fat, tendon, lung, liver, muscle, marrow stroma, and even brain cells. Conversely, neural stem cells as well as progenitors present in the muscle may contribute to blood cell production, indicating that adult stem cells present in numerous tissues may generate multiple cell types even of different dermal origin. Therefore, the developmental potential of adult somatic stem cells might be reassessed, although the mechanisms that ultimately lead to determination of cell fate are not completely defined. The successful long-term culturing and expansion of somatic adult stem cells together with their intrinsic versatility leads to future hope of stem cell therapeutic use in a wide spectrum of diseases and disorders of several, even not easily accessible, tissues.

The results here presented regard four seasonal multidisciplinary surveys conducted in the Southern Adriatic basin, in the framework of the collaborative research program PRISMA (Programma di Ricerca e Sperimentazione del Mare Adriatico), with the aim of investigating the longitudinal fluxes across a series of significant transverse sections. The thermohaline properties of the main water masses and their seasonal and spatial variability over an entire annual cycle were analyzed. Five water masses take part in these processes; namely the Adriatic Surface Water (ASW), the Ionian Surface Water (ISW), the Levantine Intermediate Water (LIW), the Adriatic Deep Water (ADW), which overflows the Sill of Otranto to form the main component of the Eastern Mediterranean Deep Water, and finally the Northern Adriatic Dense Water (NADW). In order to follow the spatial extension of these water masses and their seasonal variability, the distribution of thermohaline properties along the transverse sections were considered. The major results concern the advection of the NADW, which has been observed to flow southward over the Pelagosa Sill and subsequently into the southernmost western shelf area, reaching the Otranto Strait. The horizontal thermohaline fields, objectively analyzed, show the main pathways of the above-mentioned water masses. Important signals in the seasonal variability of the baroclinic basin-wide circulation have been noted. Finally, a quantitative measure of the cross-sectional areas occupied by the intermediate and deep waters along the transverse sections located at the Gargano passage and Otranto Strait have been calculated and are discussed.

Abstract 2350 The finding that the epigenome of differentiated cells can be reset to a pluripotent state indicates that any somatic cell may potentially change or reverse its already established developmental identity through the delivery of appropriate instructive signals. Here we show the possibility of generating mature and functional neurons from Cord Blood (CB) cells without reprogramming into a pluripotent state. In particular, we demonstrate that the ectopic expression of two transcription factors (SOX2, c-MYC) as well as only one factor (SOX2) allows the generation of proliferating neural progenitor cells starting from CB CD133+ cells. Given its important role in neural stem cell activity, we first tested if only SOX2 was sufficient to induce the conversion of CB cells into neural progenitor cells. Three weeks after retrovirus infection, we observed the formation of a few colonies showing an iPS morphology, although they were negative for pluripotency markers (OCT4, NANOG, SSEA4). However, these colonies homogenously expressed multiple neural markers (TJU1, GFAP, MAP2, PAX6, NF) and we called them CB-inducible neural cells (CB-iNCs). Since, it is known that c-MYC has a role in controlling self-renewal and proliferation of neural progenitor cells, we therefore tested the neuron-inducing activity of SOX2 in combination with c-MYC. The data showed that the presence of c-MYC increased the efficiency 20 fold. In addition, we have demonstrated that CB-iNCs were able to differentiate into functional mature neurons in vitro and in vivo. The expression of the mature synaptic marker, synapsin, and both excitatory (VGLUT-1) as well as inhibitory markers (GABA), indicated that CB-derived neurons had the protein machinery necessary to fire action potentials. To investigate if CB-derived neurons had functional membrane properties similar to neurons, we performed patch-clamp recording and calcium transient assays. The results confirmed that CB-derived neurons could form functional synapses and generate action potentials. Finally, an in vivo assay, where CB-derived neurons were injected into dentate girus of mice hippocampus, demonstrated that these cells were able to engraft, differentiate as well as extend processes along the corpus callosum, one month after transplantation. Next we compared the global gene expression analysis of CB CD133+ cells, CB-iNCs and CB-derived mature neurons. The results indicated that CB-derived neurons have a neural transcription profile similar to neurons derived from human ES. In addition, to gain further insight into the role of SOX2 during the conversion process, we investigated whether genes up-regulated in CB-iNCs, which are known to have potential SOX2 binding sites were also bound by SOX2 in our CB-iNCs. Using Chromatin immunoprecipitation (ChIP) we found evidence that SOX2 was bound to NEUROD1, DCX, NAV2, MASH1 and CHD7 genes. All these data suggest that the ectopic expression of SOX2 and c-MYC as well as only SOX2 can rapidly convert CB cells into function neurons. The possibility to generate functional neurons starting from CB cells, in an efficient and easy way, could offer a novel and powerful system for studying human cellular identity and plasticity. Disclosures: No relevant conflicts of interest to declare.

The main limitations of CB transplantation (CBT) are represented, in comparison to bone marrow transplantation, by a lower probability of engraftment and a slower hematopoietic recovery which can favor the occurrence of fatal complications. Several studies have clearly documented that the higher the number of cells infused, the faster the rate of engraftment and the lower the risk of transplant-related mortality. In order to increase the number of CB cells infused, approaches based on ex vivo expansion of CB progenitor cells are, in principle, particularly promising. In this regard, we developed a clinical protocol to expand the number of CB hematopoietic progenitor cells (HPCs), based on the use of a cytokine cocktail including interleukin-6 (10 ng/ml), stem cell factor (50 ng/ml), Flt-3 ligand (50 ng/ml) and thrombopoietin (10 ng/ml), in the presence of a serum-free medium and 10% AB allogeneic plasma. This combination induced significant and sustained ex vivo expansion of primitive CB HPCs. After clinical grade immunoselection (CliniMACS, Miltenyi Biotech), CD34+ cells were cultured at a concentration of 5x103 cells/ml for 2 weeks. Cytokines were added at the onset of culture and replaced twice a week. The purity of CD34+ cells (median and range) after immunoselection of thawed CB samples was 79.5% (47–96). After culture, the median fold expansion of total nucleated cells (NC) and CFU-GM was around 2 logs, while that of CD34+ cells was around 1 log. The expanded cells had levels of endotoxin below the required limit of 0.05IU/mL (E.Ph.) and were negative for mycoplasma, bacterial and fungal contamination. Two children, one with thalassemia and one with sickle cell disease (SCD), were given ex vivo expanded CD34+ cells from an HLA-identical sibling. In both cases the conditioning regimen included busulfan, thiotepa and fludarabine, while GVHD prophylaxis consisted of cyclosporine-A. The first patient, a 6-year-old boy with thalassemia, was given 2 CB units derived from 2 HLA-identical fraternal (i.e. non syngeneic) twins (1 unmanipulated CB unit and 1 CB unit infused after ex vivo expansion). The fold expansions of NC and CD34+ cells were 623 and 56.6 respectively. The patient had neutrophil recovery (i.e. above 500/cmm) on day +16, while the platelet recovery (i.e. above 30,000/cmm) occurred on day +31. The second patient, a 7-year-old girl with SCD, was given 2/3 of the CB unit unmanipulated and 1/3 after ex vivo expansion. The fold expansions of NC and CD34+ cells were 189.3 and 30.5 respectively. Neutrophil and platelet recovery occurred on day +18 and +33 respectively. Neither acute nor chronic GVHD occurred in both patients, who are alive and disease free at 12 and 5 months after CBT respectively. In the first patient, chimerism evaluation showed the presence of erythroid and myeloid colonies, as well as of T cells, derived from the ex vivo expanded cells up to 7 months after CBT. Mesenchymal stem cells remained of patient origin in both cases. These preliminary results indicate that our expansion procedure is safe and able to ex vivo expand CB HPCs; the expanded cells seem to accelerate hematopoietic recovery and, for the first time, we demonstrate that these cells contribute to sustain hematopoietic recovery after CBT. Altogether these results could widen the applicability of CBT in adults.

There are experimental evidences that Endothelial Progenitor Cells (EPCs) is a pool of cells that could proliferate and differentiate into mature endothelial cells and that may contribute to endothelial repair, but no systematic studies exist regarding physiologic variations in EP circulating pool and in CFU-Endo capacity in healthy subjects. In this regard, since the level of EPCs may be a biologic marker for vascular function, we decided to characterize primitive and circulating EPCs and their relation with gender, aging and the capacity to form CFU-Endo in a selected healthy population. Each subject underwent a screening evaluation to exclude any underlying cardiac diseases. A 20mL sample of peripheral blood was obtained (h 11:00–12:00 AM) from 15 male and 17 female age matched subjects (mean age 41.6±15.9yrs). EPCs were assessed by flow cytometric analysis on 100,000 events per sample (FACScan, Becton Dickinson). Primitive EPCs, defined as CD45−/34+/133+ and CD45−/KDR+/144−, were 8±1.8SE/uL. Circulating EPCs, defined as CD45−/146+/31+/FCSlow and CD45−/146+/144+, were 18.72±2.5SE/uL. No correlation was found between these populations and aging, gender and BMI. Instead a significant decrease in CD45−/34+/133+ (2.9±0.6SE/uL) subpopulation was observed with aging (r = 0.45; p = 0.009). CFU-Endo were performed as described by Hill JM et al. (NEJM 2003) and scored by microscopy in blind by two operators. The confirmation of endothelial-cell lineage was performed by immunostaining for endothelial-specific antibodies (CD31+, CD105+, CD144+, CD146low, CD133−, vWf-). CFU-Endo were expressed as the mean number of colonies per well in 8–25 separate determinations for each subjects and only 15 subjects gave colonies (mean 2.1±0.05SE). CFU-Endo do not correlate with aging and gender in healthy subjects confirming that endothelial injuries and cardiovascular risk factors are a crucial endothelium stimulus. In fact, recently it has been reported that higher colony counts were found in subjects with various degrees of cardiovascular risks. In conclusion CD45-/34+/133+ cell population correlates with aging but since this population represents Hemato-Endo Precursors that include a majority of endothelial progenitors and a small number of hematopoietic stem cells, further studies will be needed to identify a subpopulation strongly predictive of CFU-Endo capacity and to assess its endothelial lineage switch.

Hydrographic measurements obtained in March–April 2002 indicate a new ventilation of the deep layers in the Adriatic and Ionian Seas, following abrupt changes in the thermohaline circulation of the eastern Mediterranean observed since the beginning of the last decade. The water masses that reside in the Southern Adriatic basin were renewed by open ocean winter convection of water with potential density excess (óè) > 29.18 kg m–3 at depths 600 to 800 m and by a vein of still denser (29.26 kg m–3) and oxygen-rich (230 to 232 mmol m–3) water flowing into the southern Adriatic depression (~1200 m), presumably from the north. When compared to 1999, a moderate increase in salinity (~0.05) and a much stronger dissolved oxygen increase (>29 mmol m–3) were found in the bottom layer of the southern Adriatic Sea. The cold, fresh and highly oxygenated water of Adriatic origin, overflowing the Otranto Strait sill (~935 m) with a potential density of 29.24 kg m–3, was dense enough to sink into the deep Ionian basin. Multiparameter analysis based on the fundamentals of the mixing processes was applied using temperature, salinity, oxygen and nutrient data to investigate the spatial distribution of water masses and to quantify the fractional contributions of distinctive source water types along specific sections running from the Adriatic to the Ionian Sea. The results clearly indicate that the dense water of Adriatic origin, which was recently in contact with the atmosphere, have again replenished the volume of the deep Ionian basin by more than 50%.

Gamete failure-derived infertility affects millions of people worldwide; for many patients, gamete donation by unrelated donors is the only available treatment. Embryonic stem cells (ESCs) can differentiate in vitro into germ-like cells, but they are genetically unrelated to the patient. Using an in vitro protocol that aims at recapitulating development, we have achieved, for the first time, complete differentiation of human induced pluripotent stem cells (hiPSCs) to postmeiotic cells. Unlike previous reports using human ESCs, postmeiotic cells arose without the over-expression of germline related transcription factors. Moreover, we consistently obtained haploid cells from hiPSCs of different origin (keratinocytes and cord blood), produced with a different number of transcription factors, and of both genetic sexes, suggesting the independence of our approach from the epigenetic memory of the reprogrammed somatic cells. Our work brings us closer to the production of personalized human gametes in vitro.