Kdm7a expression is spatiotemporally regulated in developing Xenopus laevis embryos, and its overexpression influences late retinal development.

BACKGROUND: Post-translational histone modifications are among the most common epigenetic modifications that orchestrate gene expression, playing a pivotal role during embryonic development and in various pathological conditions. Among histone lysine demethylases, KDM7A, also known as KIAA1718 or JHDM1D, catalyzes the demethylation of H3K9me1/2 and H3K27me1/2, leading to transcriptional regulation. Previous data suggest that KDM7A plays a central role in several biological processes, including cell proliferation, commitment, differentiation, apoptosis, and maintenance. However, information on the expression pattern of KDM7A in whole organisms is limited, and its functional role is still unclear. RESULTS: In Xenopus development, kdm7a is expressed early, undergoing spatiotemporal regulation in various organs and tissues, including the central nervous system and the eye. Focusing on retinal development, we found that kdm7a overexpression does not affect the expression of genes critically involved in early neural development and eye-field specification, whereas unbalances the distribution of neural cell subtypes in the mature retina by disfavoring the development of ganglion cells while promoting that of horizontal cells. CONCLUSIONS: Kdm7a is dynamically expressed during embryonic development, and its overexpression influences late retinal development, suggesting a potential involvement in the molecular machinery regulating the spatiotemporally ordered generation of retinal neuronal subtypes.

dCas9-Based Scn1a Gene Activation Restores Inhibitory Interneuron Excitability and Attenuates Seizures in Dravet Syndrome Mice.

Dravet syndrome (DS) is a severe epileptic encephalopathy caused mainly by heterozygous loss-of-function mutations of the SCN1A gene, indicating haploinsufficiency as the pathogenic mechanism. Here we tested whether catalytically dead Cas9 (dCas9)-mediated Scn1a gene activation can rescue Scn1a haploinsufficiency in a mouse DS model and restore physiological levels of its gene product, the Nav1.1 voltage-gated sodium channel. We screened single guide RNAs (sgRNAs) for their ability to stimulate Scn1a transcription in association with the dCas9 activation system. We identified a specific sgRNA that increases Scn1a gene expression levels in cell lines and primary neurons with high specificity. Nav1.1 protein levels were augmented, as was the ability of wild-type immature GABAergic interneurons to fire action potentials. A similar enhancement of Scn1a transcription was achieved in mature DS interneurons, rescuing their ability to fire. To test the therapeutic potential of this approach, we delivered the Scn1a-dCas9 activation system to DS pups using adeno-associated viruses. Parvalbumin interneurons recovered their firing ability, and febrile seizures were significantly attenuated. Our results pave the way for exploiting dCas9-based gene activation as an effective and targeted approach to DS and other disorders resulting from altered gene dosage.

Climate warming restructures an aquatic food web over 28 years.

Climate warming can restructure lake food webs if trophic levels differ in their thermal responses, but evidence for these changes and their underlying mechanisms remain scarce in nature. Here we document how warming lake temperatures by up to 2°C, rather than changes in trophic state or fishing effort, have restructured the pelagic food web of a large European lake (Lake Maggiore, Italy). Our approach exploited abundance and biomass data collected weekly to yearly across five trophic levels from 1981 to 2008. Temperature generally had stronger effects on taxa than changes in fish predation or trophic state mediated through primary productivity. Consequently, we found that, as the lake warmed, the food web shifted in numerical abundance towards predators occupying middle trophic positions. Of these taxa, the spiny water flea (Bythotrephes longimanus) most prospered. Bythotrephes strongly limited abundances of the keystone grazer Daphnia, strengthening top-down structuring of the food web. Warmer temperatures partly restructured the food web by advancing peak Bythotrephes densities by approximately 60 days and extending periods of positive population growth by three times. Nonetheless, our results suggested that advances in the timing and size of peak Bythotrephes densities could not outpace changes in the timing and size of peak densities in their Daphnia prey. Our results provide rare evidence from nature as to how long-term warming can favour higher trophic levels, with the potential to strengthen top-down control of food webs.

AAV-PHP.B-Mediated Global-Scale Expression in the Mouse Nervous System Enables GBA1 Gene Therapy for Wide Protection from Synucleinopathy.

The lack of technology for direct global-scale targeting of the adult mouse nervous system has hindered research on brain processing and dysfunctions. Currently, gene transfer is normally achieved by intraparenchymal viral injections, but these injections target a restricted brain area. Herein, we demonstrated that intravenous delivery of adeno-associated virus (AAV)-PHP.B viral particles permeated and diffused throughout the neural parenchyma, targeting both the central and the peripheral nervous system in a global pattern. We then established multiple procedures of viral transduction to control gene expression or inactivate gene function exclusively in the adult nervous system and assessed the underlying behavioral effects. Building on these results, we established an effective gene therapy strategy to counteract the widespread accumulation of α-synuclein deposits throughout the forebrain in a mouse model of synucleinopathy. Transduction of A53T-SCNA transgenic mice with AAV-PHP.B-GBA1 restored physiological levels of the enzyme, reduced α-synuclein pathology, and produced significant behavioral recovery. Finally, we provided evidence that AAV-PHP.B brain penetration does not lead to evident dysfunctions in blood-brain barrier integrity or permeability. Altogether, the AAV-PHP.B viral platform enables non-invasive, widespread, and long-lasting global neural expression of therapeutic genes, such as GBA1, providing an invaluable approach to treat neurodegenerative diseases with diffuse brain pathology such as synucleinopathies.

Acceleration of cyanobacterial dominance in north temperate-subarctic lakes during the Anthropocene.

Increases in atmospheric temperature and nutrients from land are thought to be promoting the expansion of harmful cyanobacteria in lakes worldwide, yet to date there has been no quantitative synthesis of long-term trends. To test whether cyanobacteria have increased in abundance over the past ~ 200 years and evaluate the relative influence of potential causal mechanisms, we synthesised 108 highly resolved sedimentary time series and 18 decadal-scale monitoring records from north temperate-subarctic lakes. We demonstrate that: (1) cyanobacteria have increased significantly since c. 1800 ce, (2) they have increased disproportionately relative to other phytoplankton, and (3) cyanobacteria increased more rapidly post c. 1945 ce. Variation among lakes in the rates of increase was explained best by nutrient concentration (phosphorus and nitrogen), and temperature was of secondary importance. Although cyanobacterial biomass has declined in some managed lakes with reduced nutrient influx, the larger spatio-temporal scale of sedimentary records show continued increases in cyanobacteria throughout the north temperate-subarctic regions.

Synapsin Condensates Recruit alpha-Synuclein.

Neurotransmission relies on the tight spatial and temporal regulation of the synaptic vesicle (SV) cycle. Nerve terminals contain hundreds of SVs that form tight clusters. These clusters represent a distinct liquid phase in which one component of the phase are SVs and the other synapsin 1, a highly abundant synaptic protein. Another major family of disordered proteins at the presynapse includes synucleins, most notably α-synuclein. The precise physiological role of α-synuclein in synaptic physiology remains elusive, albeit its role has been implicated in nearly all steps of the SV cycle. To determine the effect of α-synuclein on the synapsin phase, we employ the reconstitution approach using natively purified SVs from rat brains and the heterologous cell system to generate synapsin condensates. We demonstrate that synapsin condensates recruit α-synuclein, and while enriched into these synapsin condensates, α-synuclein still maintains its high mobility. The presence of SVs enhances the rate of synapsin/α-synuclein condensation, suggesting that SVs act as catalyzers for the formation of synapsin condensates. Notably, at physiological salt and protein concentrations, α-synuclein alone is not able to cluster isolated SVs. Excess of α-synuclein disrupts the kinetics of synapsin/SV condensate formation, indicating that the molar ratio between synapsin and α-synuclein is important in assembling the functional condensates of SVs. Understanding the molecular mechanism of α-synuclein interactions at the nerve terminals is crucial for clarifying the pathogenesis of synucleinopathies, where α-synuclein, synaptic proteins and lipid organelles all accumulate as insoluble intracellular inclusions.

In situ diagnostic analysis of the XVIII century Madonna della Lettera panel painting (Messina, Italy).

The scientific investigation of the techniques employed by the artist, such as composition of the paints, color palette, and painting style represents a fundamental pre-requisite in order to develop proper conservation and restoration strategies. In this context, the combined use of non-destructive, non-invasive in situ image and chemical analyses was here successfully employed for the investigation of the XVIII century Madonna della Lettera panel painting from the ancient Basilian abbey of Santa Maria di Bordonaro in Messina (Italy). The used techniques were visible, infrared (IR) and false-color infrared (FCIR) photography, X-ray radiography, X-ray fluorescence (XRF) and Raman spectroscopy. The goal was to obtain accurate information on materials and techniques originally used and possible later interventions, with particular regard to the nature of the painting materials. From the results, details of the artwork useful for restoration and conservation procedures were revealed. The identification of most of the artist's palette was also achieved: Prussian blue for blue color, lead white for white, umber for the brown, cinnabar for the red, and carbon black for the black color. The composition of different preparatory substrates was also investigated. The obtained results, other than constituting a crucial step for future restoration works, can be at the same time useful for the dating of the painting, that does not report the date and the artist's signature.

Whole brain delivery of an instability-prone Mecp2 transgene improves behavioral and molecular pathological defects in mouse models of Rett syndrome.

Rett syndrome is an incurable neurodevelopmental disorder caused by mutations in the gene encoding for methyl-CpG binding-protein 2 (MeCP2). Gene therapy for this disease presents inherent hurdles since MECP2 is expressed throughout the brain and its duplication leads to severe neurological conditions as well. Herein, we use the AAV-PHP.eB to deliver an instability-prone Mecp2 (iMecp2) transgene cassette which, increasing RNA destabilization and inefficient protein translation of the viral Mecp2 transgene, limits supraphysiological Mecp2 protein levels. Intravenous injections of the PHP.eB-iMecp2 virus in symptomatic Mecp2 mutant mice significantly improved locomotor activity, lifespan and gene expression normalization. Remarkably, PHP.eB-iMecp2 administration was well tolerated in female Mecp2 mutant or in wild-type animals. In contrast, we observed a strong immune response to the transgene in treated male Mecp2 mutant mice that was overcome by immunosuppression. Overall, PHP.eB-mediated delivery of iMecp2 provided widespread and efficient gene transfer maintaining physiological Mecp2 protein levels in the brain.

Pharmacological Inhibition of Necroptosis Protects from Dopaminergic Neuronal Cell Death in Parkinson's Disease Models.

Dysfunctions in mitochondrial dynamics and metabolism are common pathological processes associated with Parkinson's disease (PD). It was recently shown that an inherited form of PD and dementia is caused by mutations in the OPA1 gene, which encodes for a key player in mitochondrial fusion and structure. iPSC-derived neural cells from these patients exhibited severe mitochondrial fragmentation, respiration impairment, ATP deficits, and heightened oxidative stress. Reconstitution of normal levels of OPA1 in PD-derived neural cells normalized mitochondria morphology and function. OPA1-mutated neuronal cultures showed reduced survival in vitro. Intriguingly, selective inhibition of necroptosis effectively rescued this survival deficit. Additionally, dampening necroptosis in MPTP-treated mice protected from DA neuronal cell loss. This human iPSC-based model captures both early pathological events in OPA1 mutant neural cells and the beneficial effects of blocking necroptosis, highlighting this cell death process as a potential therapeutic target for PD.

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.

European environmental scenarios of chemical bioavailability in freshwater systems.

In exposure prediction for environmental risk assessment, the transition to more dynamic and realistic modelling approaches and scenarios has been recently identified as a major challenge, since it would allow a more accurate prediction of bioavailable concentrations and their variations in space and time. In this work, an improved version of the multimedia model ChimERA fate, including a phytoplankton compartment and equations to calculate phytoplankton, detritus and dissolved organic matter variations in time, was developed. The model was parameterized to simulate five dynamic scenarios for shallow meso-eutrophic water bodies based on a latitudinal gradient (in Europe); such scenarios include seasonal profiles of water temperature, phytoplankton biomass, detritus, and dissolved organic matter. Model runs were performed for each scenario for 8 hydrophobic chemicals (PCB congeners), with the aim of investigating the influence of scenario characteristics and compound properties on bioavailable concentrations. The key processes were adsorption/uptake by phytoplankton and deposition to sediment of detritus-bound chemicals. The northern scenarios ("Scandinavia" and "UK") showed the highest bioavailable concentrations, with annual maximum/minimum concentration up to 25; in contrast, for example, maximum concentrations in the "Mediterranean" scenario were lower by a factor of 2 to 9 with respect to the northern ones (depending on chemical hydrophobicity), due to the generally higher biomass and carbon levels, and showed only limited seasonal variability (up to a factor of 4). These results highlight the importance of including biomass and organic carbon dynamics in both modelling approaches and scenarios for the evaluation of exposure concentrations in aquatic environments.

Warming trends of perialpine lakes from homogenised time series of historical satellite and in-situ data.

The availability of more than thirty years of historical satellite data is a valuable source which could be used as an alternative to the sparse in-situ data. We developed a new homogenised time series of daily day time Lake Surface Water Temperature (LSWT) over the last thirty years (1986-2015) at a spatial resolution of 1km from thirteen polar orbiting satellites. The new homogenisation procedure implemented in this study corrects for the different acquisition times of the satellites standardizing the derived LSWT to 12:00 UTC. In this study, we developed new time series of LSWT for five large lakes in Italy and evaluated the product with in-situ data from the respective lakes. Furthermore, we estimated the long-term annual and summer trends, the temporal coherence of mean LSWT between the lakes, and studied the intra-annual variations and long-term trends from the newly developed LSWT time series. We found a regional warming trend at a rate of 0.017°Cyr-1 annually and 0.032°Cyr-1 during summer. Mean annual and summer LSWT temporal patterns in these lakes were found to be highly coherent. Amidst the reported rapid warming of lakes globally, it is important to understand the long-term variations of surface temperature at a regional scale. This study contributes a new method to derive long-term accurate LSWT for lakes with sparse in-situ data thereby facilitating understanding of regional level changes in lake's surface temperature.

Benefits and limitations of an intercalibration of phytoplankton assessment methods based on the Mediterranean GIG reservoir experience.

The status of European legislation regarding inland water quality after the enactment of the Water Framework Directive (WFD) originated scientific effort to develop reliable methods, primarily based on biological parameters. An important aspect of the process was to ensure that quality assessment was comparable between the different Member States. The Intercalibration process (IC), required in the WFD ensures the unbiased application of the norm. The presented results were developed in the context of the 2nd IC phase. An overview of the reservoir type definition of the Lake Mediterranean Geographical Intercalibration Group, where four types were considered divided by both alkalinity and climate, together with the results for selection of Maximum Ecological Potential sites (MEP) are presented. MEP reservoirs were selected based on pressure and biological variables. Three phytoplankton-based assessment methods were intercalibrated using data from Mediterranean countries. The Mediterranean Assessment System for Reservoirs Phytoplankton (Spain), the New Mediterranean Assessment System for Reservoirs Phytoplankton (Portugal and Cyprus) and the New Italian Method (Italy) were applied. These three methods were compared through option 3 of the Intercalibration Guide. The similarity of the assessments was quantified, and the Good/Moderate (GM) boundaries assessed. All three methods stood as comparable at the GM boundary except for the MASRP in siliceous wet reservoirs, which was slightly stricter. Finally, the main taxonomic groups represented in the phytoplankton community at MEP conditions were identified, as well as their main changes with an increasing trophic status. MEP sites are dominated by chrysophytes in siliceous wet reservoirs and by the diatoms Cyclotella and Achnanthes in calcareous ones. Cyanobacteria take over the community in both calcareous and siliceous wet reservoirs as eutrophication increases. In summary, the relevance and reliability of the quality assessment methods compared were confirmed both from an ecological perspective and a health risk management point of view.

A longitudinal EEG study of Alzheimer's disease progression based on a complex network approach.

A complex network approach is combined with time dynamics in order to conduct a space-time analysis applicable to longitudinal studies aimed to characterize the progression of Alzheimer's disease (AD) in individual patients. The network analysis reveals how patient-specific patterns are associated with disease progression, also capturing the widespread effect of local disruptions. This longitudinal study is carried out on resting electroence phalography (EEGs) of seven AD patients. The test is repeated after a three months' period. The proposed methodology allows to extract some averaged information and regularities on the patients' cohort and to quantify concisely the disease evolution. From the functional viewpoint, the progression of AD is shown to be characterized by a loss of connected areas here measured in terms of network parameters (characteristic path length, clustering coefficient, global efficiency, degree of connectivity and connectivity density). The differences found between baseline and at follow-up are statistically significant. Finally, an original topographic multiscale approach is proposed that yields additional results.

Assessing remotely sensed chlorophyll-a for the implementation of the Water Framework Directive in European perialpine lakes.

The lakes of the European perialpine region constitute a large water reservoir, which is threatened by the anthropogenic pressure altering water quality. The Water Framework Directive of the European Commission aims to protect water resources and monitoring is seen as an essential step for achieving this goal. Remote sensing can provide frequent data for large scale studies of water quality parameters such as chlorophyll-a (chl-a). In this work we use a dataset of maps of chl-a derived from over 200 MERIS (MEdium Resolution Imaging Spectrometer) satellite images for comparing water quality of 12 perialpine lakes in the period 2003-2009. Besides the different trophic levels of the lakes, results confirm that the seasonal variability of chl-a concentration is particularly pronounced during spring and autumn especially for the more eutrophic lakes. We show that relying on only one sample for the assessment of lake water quality during the season might lead to misleading results and erroneous assignments to quality classes. Time series MERIS data represents a suitable and cost-effective technology to fill this gap, depicting the dynamics of the surface waters of lakes in agreement with the evolution of natural phenomena.