Impaired Mitochondrial Function and Marrow Failure in Patients Carrying a Variant of the SRSF4 Gene.

Serine/arginine-rich splicing factors (SRSFs) are a family of proteins involved in RNA metabolism, including pre-mRNA constitutive and alternative splicing. The role of SRSF proteins in regulating mitochondrial activity has already been shown for SRSF6, but SRSF4 altered expression has never been reported as a cause of bone marrow failure. An 8-year-old patient admitted to the hematology unit because of leukopenia, lymphopenia, and neutropenia showed a missense variant of unknown significance of the SRSF4 gene (p.R235W) found via whole genome sequencing analysis and inherited from the mother who suffered from mild leuko-neutropenia. Both patients showed lower SRSF4 protein expression and altered mitochondrial function and energetic metabolism in primary lymphocytes and Epstein-Barr-virus (EBV)-immortalized lymphoblasts compared to healthy donor (HD) cells, which appeared associated with low mTOR phosphorylation and an imbalance in the proteins regulating mitochondrial biogenesis (i.e., CLUH) and dynamics (i.e., DRP1 and OPA1). Transfection with the wtSRSF4 gene restored mitochondrial function. In conclusion, this study shows that the described variant of the SRSF4 gene is pathogenetic and causes reduced SRSF4 protein expression, which leads to mitochondrial dysfunction. Since mitochondrial function is crucial for hematopoietic stem cell maintenance and some genetic bone marrow failure syndromes display mitochondrial defects, the SRSF4 mutation could have substantially contributed to the clinical phenotype of our patient.

Anthropogenic contribution, transport, and accumulation of Polycyclic Aromatic Hydrocarbons in sediments of the continental shelf and slope in the Mediterranean Sea.

A multidisciplinary approach, involving geochemical, sedimentological and oceanographic analyses, was employed to examine the distribution of Polycyclic Aromatic Hydrocarbons (PAHs) in a strongly anthropized area of the marginal Adriatic Sea (Mediterranean basin). The investigation into PAH distribution considered the grain size and biogeochemical properties of the sediments, as well as in relation to the main oceanographic processes and river inputs. Both biogeochemical and hydrographical inputs regulated the sedimentation of organic particles, influencing the distribution of PAHs. The results indicated PAH levels in 116 marine surface sediments ranging from 4 to 235 ng g-1 (average 55 ng g-1). The distribution of PAHs in Adriatic Sea surface sediments aligned with a higher clayey sedimentation in the deeper basin areas of the Middle Adriatic Depression.

Fecal bacteria contamination in the Adriatic Sea: Investigating environmental factors and modeling to manage recreational coastal waters.

This study is based on assessing fecal indicator bacteria contamination along meteorological, hydrological and physical-chemical variables after high rainy events during the summer period. The study focused on four different coastal sites in the western and eastern Adriatic coast characterized by various geomorphological and hydrological features, levels of urbanization and anthropogenic pressures, with the aim of finding appropriate and effective solutions to ensure the safety and sustainability of tourism and public health. Detailed in-situ survey revealed a wide range of fecal indicator bacterial (FIB) across the different river mouths with concentrations of E. coli ranging from 165 to 6700 CFU 100 mL-1. It was found that nitrogen compounds track microbial load and acted as tracers for fecal contaminants. Further, a modelling tool was also used to analyze the spatial and temporal distribution of fecal pollution at these coastal sites. The integrated monitoring through high frequent survey in river waters and modeling framework allowed for the estimation of fecal indicator bacterial load at the river mouth and examination of fecal pollutant dispersion in recreational waters, considering different scenarios of fecal dispersion along the coast. This study formed the basis of a robust decision support system aimed at improving the management of recreational areas and ensuring the protection of water bodies through efficient management of bathing areas.

Microstructured Polymeric Fabrics Modulating the Paracrine Activity of Adipose-Derived Stem Cells.

The deposition of stem cells at sites of injury is a clinically relevant approach to facilitate tissue repair and angiogenesis. However, insufficient cell engraftment and survival require the engineering of novel scaffolds. Here, a regular network of microscopic poly(lactic-co-glycolic acid) (PLGA) filaments was investigated as a promising biodegradable scaffold for human Adipose-Derived Stem Cell (hADSC) tissue integration. Via soft lithography, three different microstructured fabrics were realized where 5 × 5 and 5 × 3 µm PLGA 'warp' and 'weft' filaments crossed perpendicularly with pitch distances of 5, 10 and 20 µm. After hADSC seeding, cell viability, actin cytoskeleton, spatial organization and the secretome were characterized and compared to conventional substrates, including collagen layers. On the PLGA fabric, hADSC re-assembled to form spheroidal-like structures, preserving cell viability and favoring a nonlinear actin organization. Moreover, the secretion of specific factors involved in angiogenesis, the remodeling of the extracellular matrix and stem cell homing was favored on the PLGA fabric as compared to that which occurred on conventional substrates. The paracrine activity of hADSC was microstructure-dependent, with 5 µm PLGA fabric enhancing the expression of factors involved in all three processes. Although more studies are needed, the proposed PLGA fabric would represent a promising alternative to conventional collagen substrates for stem cell implantation and angiogenesis induction.

Accounting for environmental and fishery management factors when standardizing CPUE data from a scientific survey: A case study for Nephrops norvegicus in the Pomo Pits area (Central Adriatic Sea).

Abundance and distribution of commercial marine resources are influenced by environmental variables, which together with fishery patterns may also influence their catchability. However, Catch Per Unit Effort (CPUE) can be standardized in order to remove most of the variability not directly attributable to fish abundance. In the present study, Generalized Additive Models (GAMs) were used to investigate the effect of some environmental and fishery covariates on the spatial distribution and abundance of the Norway lobster Nephrops norvegicus within the Pomo/Jabuka Pits (Central Adriatic Sea) and to include those that resulted significant in a standardization process. N. norvegicus is a commercially important demersal crustacean, altering its catchability over the 24-h cycle and seasons according to its burrowing behavior. A historically exploited fishing ground for this species, since 2015 subject to specific fisheries management measures, is represented by the meso-Adriatic depressions, which are also characterized by particular oceanographic conditions. Both the species behaviour and the features of this study area influence the dynamics of the population offering a challenging case study for a standardization modelling approach. Environmental and catch data were obtained during scientific trawl surveys properly designed to catch N. norvegicus, thus improving the quality of the model input data. Standardization of CPUE from 2 surveys from 2012 to 2019 was conducted building two GAMs for both biomass and density indices. Bathymetry, fishing pressure, dissolved oxygen and salinity proved to be significant drivers influencing catch distribution. After cross validations, the tuned models were then used to predict new indices for the study area and the two survey series by means of informed spatial grids, composed by constant surface cells, to each of which are associated average values of environmental parameters and specific levels of fishing pressure, depending on the management measures in place. The predictions can be used to better describe the structure and the spatio-temporal distribution of the population providing valuable information to evaluate the status of such an important marine resource.

Assessment of seasonal relationship between polycyclic aromatic hydrocarbon accumulation and expression patterns of oxidative stress-related genes in muscle tissues of red mullet (M. barbatus) from the Northern Adriatic Sea.

In this study, we examined the seasonal association between Polycyclic Aromatic Hydrocarbon (PAH) concentrations and mRNA expression profiles of some antioxidant genes (i.e. CAT, GST and SOD), as well as lipid peroxidation (LPO), in muscle of sexually inactive females of red mullet (Mullus barbatus). Fish were captured in a fishery area of the Northern Adriatic Sea during both winter and summer. We found significantly (p < 0.05) higher ∑HMW-PAHs concentrations in muscle of specimens caught during winter than summer. On the basis of sampling season, red mullets exhibited different gene expression profiles of antioxidant enzymes showing lower levels of both CAT and GST in winter than in summer. Accordingly, CAT was found to be negatively associated with ∑PAH concentrations, especially ∑LMW-PAH, in individuals collected during winter. Seasonal-related downregulation of some oxidative stress biomarker expression is suggestive of greater susceptibility of red mullets to PAHs during winter.

Water quality integrated system: A strategic approach to improve bathing water management.

In the Adriatic Sea, massive rainfall events are causing flooding of rivers and streams, with severe consequences on the environment. The consequent bacterial contamination of bathing water poses public health risks besides damaging tourism and the economy. This study was conducted in the framework of WATERCARE, an EU Interreg Italy-Croatia Project, which aims at reducing the impact of microbial contamination on Adriatic bathing water due to heavy rainfall events drained in the local sewage network and; enhancing the quality of local waters; and providing support for the decision-making processes regarding the management of bathing water in line with EU regulations. The study involved the development of an innovative water quality integrated system that helps meet these objectives. It consists of four components: a real time hydro-meteorological monitoring system; an autosampler to collect freshwater samples during and after significant rainfall events; a forecast system to simulate the dispersion of pollutants in seawater; and a real-time alert system that can predict the potential ecological risk from the microbial contamination of seawater. The system was developed and tested at a pilot site (Fano, Italy). These preliminary results will be used to develop guidelines for urban wastewater and coastal system quality assessments to contribute to develop policy actions and final governance decisions.

Status of faecal pollution in ports: A basin-wide investigation in the Adriatic Sea.

Ports are subject to a variety of anthropogenic impacts, and there is mounting evidence of faecal contamination through several routes. Yet, little is known about pollution in ports by faecal indicator bacteria (FIB). FIB spatio-temporal dynamics were assessed in 12 ports of the Adriatic Sea, a semi-enclosed basin under strong anthropogenic pressure, and their relationships with environmental variables were explored to gain insight into pollution sources. FIB were abundant in ports, often more so than in adjacent areas; their abundance patterns were related to salinity, oxygen, and nutrient levels. In addition, a molecular method, quantitative (q)PCR, was used to quantify FIB. qPCR enabled faster FIB determination and water quality monitoring that culture-based methods. These data provide robust baseline evidence of faecal contamination in ports and can be used to improve the management of routine port activities (dredging and ballast water exchange), having potential to spread pathogens in the sea.

Phytoplankton diversity in Adriatic ports: Lessons from the port baseline survey for the management of harmful algal species.

An inventory of phytoplankton diversity in 12 Adriatic ports was performed with the port baseline survey. Particular emphasis was put on the detection of harmful aquatic organisms and pathogens (HAOP) because of their negative impact on ecosystem, human health, and the economy. Phytoplanktonic HAOP are identified as species, either native or non-indigenous (NIS), which can trigger harmful algal blooms (HAB). A list of 691 taxa was prepared, and among them 52 were classified as HAB and five as NIS. Records of toxigenic NIS (Pseudo-nitzschia multistriata, Ostreopsis species including O. cf. ovata) indicate that the intrusion of non-native invasive phytoplankton species has already occurred in some Adriatic ports. The seasonal occurrence and abundance of HAOP offers a solid baseline for a monitoring design in ports in order to prevent ballast water uptake and possible expansion of HAOP outside their native region.

Oceanographic characteristics of the Adriatic Sea - Support to secondary HAOP spread through natural dispersal.

Vessels, specifically ballast water and hull fouling, are a major vector for the introduction of non-indigenous species (NIS) in European seas. The Mediterranean is one of the world's marine regions where their invasion is heaviest. The shallow Adriatic basin is a highly sensitive area that is already experiencing its consequences. The secondary spread of NIS over a wider area through natural dispersion is a complex process that depends on a wide range of oceanographic factors. This work analysed the dataset of the BALMAS project, in whose framework twelve ports in the Adriatic Sea were subjected to a Port Baseline Survey (PBS), to estimate the natural spread of NIS organisms from their port of arrival to the wider Adriatic basin. Its findings indicate that the prevailing water circulation patterns facilitate the natural dispersal of harmful aquatic organisms and pathogens (HAOP).

Strategy of port baseline surveys (PBS) in the Adriatic Sea.

Port baseline surveys (PBS) provide species inventories in and around ports, with a focus on non-indigenous species that may have been introduced by vessels, primarily via ballast water. PBS are an essential tool to support effective management strategies for non-indigenous as well as native harmful aquatic organisms and pathogens (HAOP). This paper describes the methodology of PBS that were conducted in 12 Adriatic ports. The PBS employed existing protocols that were adapted to meet the characteristics of the Adriatic sites. Their results are reported in several papers included in this special issue, each of which is devoted to a specific community. An overview of existing surveys protocols - which provide valuable support to decision-making and to design effective monitoring of non-indigenous species - is also supplied.

Plankton dynamics across the freshwater, transitional and marine research sites of the LTER-Italy Network. Patterns, fluctuations, drivers.

A first synoptic and trans-domain overview of plankton dynamics was conducted across the aquatic sites belonging to the Italian Long-Term Ecological Research Network (LTER-Italy). Based on published studies, checked and complemented with unpublished information, we investigated phytoplankton and zooplankton annual dynamics and long-term changes across domains: from the large subalpine lakes to mountain lakes and artificial lakes, from lagoons to marine coastal ecosystems. This study permitted identifying common and unique environmental drivers and ecological functional processes controlling seasonal and long-term temporal course. The most relevant patterns of plankton seasonal succession were revealed, showing that the driving factors were nutrient availability, stratification regime, and freshwater inflow. Phytoplankton and mesozooplankton displayed a wide interannual variability at most sites. Unidirectional or linear long-term trends were rarely detected but all sites were impacted across the years by at least one, but in many case several major stressor(s): nutrient inputs, meteo-climatic variability at the local and regional scale, and direct human activities at specific sites. Different climatic and anthropic forcings frequently co-occurred, whereby the responses of plankton communities were the result of this environmental complexity. Overall, the LTER investigations are providing an unparalleled framework of knowledge to evaluate changes in the aquatic pelagic systems and management options.

Downregulation of glutamic acid decarboxylase in Drosophila TDP-43-null brains provokes paralysis by affecting the organization of the neuromuscular synapses.

Amyotrophic lateral sclerosis is a progressive neurodegenerative disease that affects the motor system, comprised of motoneurons and associated glia. Accordingly, neuronal or glial defects in TDP-43 function provoke paralysis due to the degeneration of the neuromuscular synapses in Drosophila. To identify the responsible molecules and mechanisms, we performed a genome wide proteomic analysis to determine differences in protein expression between wild-type and TDP-43-minus fly heads. The data established that mutant insects presented reduced levels of the enzyme glutamic acid decarboxylase (Gad1) and increased concentrations of extracellular glutamate. Genetic rescue of Gad1 activity in neurons or glia was sufficient to recuperate flies locomotion, synaptic organization and glutamate levels. Analogous recovery was obtained by treating TDP-43-null flies with glutamate receptor antagonists demonstrating that Gad1 promotes synapses formation and prevents excitotoxicity. Similar suppression of TDP-43 provoked the downregulation of GAD67, the Gad1 homolog protein in human neuroblastoma cell lines and analogous modifications were observed in iPSC-derived motoneurons from patients carrying mutations in TDP-43, uncovering conserved pathological mechanisms behind the disease.

Mucilaginous aggregates in the northern Adriatic in the period 1999-2002: typology and distribution.

The spatial and temporal distributions of different types of visible mucilaginous aggregates were investigated monthly by underwater video cameras in the northern Adriatic (NA) from June 1999 to July 2002. Small aggregates (flocs, macroflocs and stringers) were observed in all seasons with higher values in autumn and winter. Larger aggregates (ribbons, cobwebs, clouds, and false bottoms) formed only during late-spring and summer, particularly in 2000 and 2002, when the seasonal thermohaline stratification was marked. In fact, the process of mucilage formation took place mainly in the upper water column, above the main pycnocline. The spatial distribution shows that higher concentrations of small aggregates occur in the more productive coastal waters. The larger aggregates form or accumulate mainly in coastal waters in the southern part of the NA and in the central part of the Po River delta-Rovinj transect, where a gyre greatly influences the aggregates distribution. The fronts between low salinity coastal waters and high salinity waters of southern origin play an important role for accumulation and aggregation of the mucilage during spring and summer. Most of the aggregates accumulate in correspondence with strong pycnoclines with differences in density anomaly of 2 kg m(-3) or higher. False bottoms formed in correspondence with strong haloclines (DeltaS congruent with 2), while cobwebs and ribbons also occurred when the halocline was less marked (DeltaS<0.5). Meteorological conditions greatly influenced the aggregate formation. Calm weather and weak winds favour aggregation processes, while strong north-easterly winds, causing intense mixing, result in the dispersion of the mucilaginous aggregates, as occurred during the Bora event in July 2000.

Circulation and horizontal fluxes in the northern Adriatic Sea in the period June 1999-July 2002. Part II: nutrients transport.

Nutrient concentrations (orthophosphate, orthosilicate and dissolved inorganic nitrogen-DIN) were measured on three transects in the northern Adriatic approximately monthly in the period June 1999-July 2002. The net nutrient transports across the three transects were estimated from these concentration values and calculated water fluxes [Grilli F, Paschini E, Russo A, Precali R, Supic N. Circulation and horizontal fluxes in the Northern Adriatic Sea in the period June 1999_July 2002. Part I: geostrophic circulation and current measurement. Sci Total Environ 2005-this issue], with the aim to verify their possible role in the mucilage phenomenon. The nutrient transports in the northern Adriatic were very variable, seasonally and from year to year, both in intensity and direction. Some differences were noticed also among the nutrient species. At the northernmost transect Po Delta-Rovinj minimal values and variable directions were recorded, probably due to the prevailing of eastward transversal transports, observed already in late winter, and generally dominant in spring and summer. Northward transport was often measured at the central transect Cesenatico-Cape Kamenjak, particularly in spring 2000 and 2002, and summer 2001, as well as for only DIN in summer 1999. In contrast, southward directions prevailed at the southernmost transect Senigallia-Susak Island. In October significant southward transports occurred at all transects and for all nutrients, particularly strong in 2000, when exceptionally high Po River discharges occurred, and the Western Adriatic Current (WAC) was well developed. However, the nutrient transports in the upper water column were low or northwards at both transect Cesenatico-Cape Kamenjak and Senigallia-Susak Island in June and July 2000, 2001, and 2002, when the mucilage phenomenon developed, but were opposite in 1999, a year without events. The results suggest that high variations of nutrient fluxes and their ratios, in conditions of reduced water dynamics, are essential for the development of the phenomenon, rather than the absolute amounts of the nutrient inputs.

Circulation and horizontal fluxes in the northern Adriatic Sea in the period June 1999-July 2002. Part I: geostrophic circulation and current measurement.

The dramatic increase in the occurrence of massive mucilage events in the northern Adriatic (NA) since their recent conspicuous reappearance in the late 1980s prompted a study of circulation and horizontal fluxes. Three transects with equidistant stations (10 km) were thus monitored monthly between June 1999 and July 2002. The geostrophic method was used to compute currents across the three transects from the CTD data, and dynamic heights provided a picture of the horizontal surface circulation. Currentmeter data records were used to adjust the reference surface and to validate the results for the southernmost and deeper (up to 70 m) transect (Senigallia-Susak Island). Geostrophic currents allowed estimation of monthly water fluxes across the transect. Different circulation regimes in the NA were observed, which may have affected mucilage events. When mucilage was absent (1999) or reduced (2001) in the western sector, the Western Adriatic Current (WAC, carrying water out of the NA) was found to be active, whilst the WAC was very weak or reversed when massive mucilage events occurred (2000 and 2002). Opposite behaviour has been observed for the Istrian Coastal Counter-Current (ICCC, retaining freshwater water in the NA) which was more intense during or after massive mucilage events and did not appear when mucilage was absent. Both WAC weakening and ICCC strengthening indicate a longer residence time of riverine waters in the NA, which favours mucilage development. Conclusively, WAC and ICCC result as key elements in controlling massive mucilage phenomena in the NA.