Toll-like receptors and IL-7 as potential biomarkers for immune-mediated necrotizing myopathies.

We aimed to verify whether the immune system may represent a source of potential biomarkers for the stratification of immune-mediated necrotizing myopathies (IMNMs) subtypes. A group of 22 patients diagnosed with IMNM [7 with autoantibodies against signal recognition particle (SRP) and 15 against 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGCR)] and 12 controls were included. A significant preponderance of M1 macrophages was observed in both SRP+ and HMGCR+ muscle samples (p < 0.0001 in SRP+ and p = 0.0316 for HMGCR+ ), with higher values for SRP+ (p = 0.01). Despite the significant increase observed in the expression of TLR4 and all endosomal Toll-like receptors (TLRs) at protein level in IMNM muscle tissue, only TLR7 has been shown considerably upregulated compared to controls at transcript level (p = 0.0026), whereas TLR9 was even decreased (p = 0.0223). Within IMNM subgroups, TLR4 (p = 0.0116) mRNA was significantly increased in SRP+ compared to HMGCR+ patients. Within IMNM group, only IL-7 was differentially expressed between SRP+ and HMGCR+ patients, with higher values in SRP+ patients (p = 0.0468). Overall, innate immunity represents a key player in pathological mechanisms of IMNM. TLR4 and the inflammatory cytokine IL-7 represent potential immune biomarkers able to differentiate between SRP+ and HMGCR+ patients.

Dysregulation of Muscle-Specific MicroRNAs as Common Pathogenic Feature Associated with Muscle Atrophy in ALS, SMA and SBMA: Evidence from Animal Models and Human Patients.

Motor neuron diseases (MNDs) are neurodegenerative disorders characterized by upper and/or lower MN loss. MNDs include amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), and spinal and bulbar muscular atrophy (SBMA). Despite variability in onset, progression, and genetics, they share a common skeletal muscle involvement, suggesting that it could be a primary site for MND pathogenesis. Due to the key role of muscle-specific microRNAs (myomiRs) in skeletal muscle development, by real-time PCR we investigated the expression of miR-206, miR-133a, miR-133b, and miR-1, and their target genes, in G93A-SOD1 ALS, Δ7SMA, and KI-SBMA mouse muscle during disease progression. Further, we analyzed their expression in serum of SOD1-mutated ALS, SMA, and SBMA patients, to demonstrate myomiR role as noninvasive biomarkers. Our data showed a dysregulation of myomiRs and their targets, in ALS, SMA, and SBMA mice, revealing a common pathogenic feature associated with muscle impairment. A similar myomiR signature was observed in patients' sera. In particular, an up-regulation of miR-206 was identified in both mouse muscle and serum of human patients. Our overall findings highlight the role of myomiRs as promising biomarkers in ALS, SMA, and SBMA. Further investigations are needed to explore the potential of myomiRs as therapeutic targets for MND treatment.

Aging-associated genes and let-7 microRNAs: a contribution to myogenic program dysregulation in oculopharyngeal muscular dystrophy.

Oculopharyngeal muscular dystrophy (OPMD) is a late-onset muscle disease caused by an abnormal (GCN) triplet expansion within the polyadenylate-binding protein nuclear 1 gene and consequent mRNA processing impairment and myogenic defects. Because a reduced cell proliferation potential and the consequent regeneration failure of aging muscle have been shown to be governed by lethal-7 (let-7) microRNA-mediated mechanisms, in the present study, we evaluated the role of let-7 in the pathogenesis of OPMD. By a multidisciplinary approach, including confocal microscopy, Western blot, and quantitative PCR analyses on muscle biopsies from patients and unaffected individuals, we found a significant increase in let-7 expression in OPMD muscles associated with an unusual high percentage of paired box 7-positive satellite cells. Furthermore, IL-6, a cytokine involved in the regulation of satellite cell proliferation and differentiation and a potential target of let-7, was found strongly down-regulated in OPMD compared with control muscles. The decrease in IL-6 transcript levels and protein content was also confirmed in vitro during differentiation of patients' and controls' muscle cells. Overall, our data suggest a key role of let-7 in the regeneration and degeneration process in OPMD muscle and pointed to IL-6 as a potential target molecule for new therapeutic approaches for this disorder.-Cappelletti, C., Galbardi, B., Bruttini, M., Salerno, F., Canioni, E., Pasanisi, M. B., Rodolico, C., Brizzi, T., Mora, M., Renieri, A., Maggi, L., Bernasconi, P., Mantegazza, R. Aging-associated genes and let-7 microRNAs: a contribution to myogenic program dysregulation in oculopharyngeal muscular dystrophy.

Zc3h10 is a novel mitochondrial regulator.

Mitochondria are the energy-generating hubs of the cell. In spite of considerable advances, our understanding of the factors that regulate the molecular circuits that govern mitochondrial function remains incomplete. Using a genome-wide functional screen, we identify the poorly characterized protein Zinc finger CCCH-type containing 10 (Zc3h10) as regulator of mitochondrial physiology. We show that Zc3h10 is upregulated during physiological mitochondriogenesis as it occurs during the differentiation of myoblasts into myotubes. Zc3h10 overexpression boosts mitochondrial function and promotes myoblast differentiation, while the depletion of Zc3h10 results in impaired myoblast differentiation, mitochondrial dysfunction, reduced expression of electron transport chain (ETC) subunits, and blunted TCA cycle flux. Notably, we have identified a loss-of-function mutation of Zc3h10 in humans (Tyr105 to Cys105) that is associated with increased body mass index, fat mass, fasting glucose, and triglycerides. Isolated peripheral blood mononuclear cells from individuals homozygotic for Cys105 display reduced oxygen consumption rate, diminished expression of some ETC subunits, and decreased levels of some TCA cycle metabolites, which all together derive in mitochondrial dysfunction. Taken together, our study identifies Zc3h10 as a novel mitochondrial regulator.

Glacier Melting Increases the Solute Concentrations of Himalayan Glacial Lakes.

Over the past two decades, we observed a substantial rise in ionic content that was mainly determined by the sulfate concentration at 20 remote high elevation lakes located in central southern Himalaya. At LCN9, which was monitored on an annual basis for the last 20 years, the sulfate concentrations increased over 4-fold. Among the main causes, we exclude a change in the composition of wet atmospheric deposition, as well as a possible influence of decrease in seasonal snow cover duration, which could have exposed larger basin surfaces to alteration processes. Glacier retreat likely was the main factor responsible for the observed increase of sulfate concentrations. We attribute this chemical changes mainly to the sulfide oxidation processes that occur in subglacial environments. Moreover, we observe that the weakened monsoon of the past two decades has only partially contributed to the lakes enrichment through runoff waters that are more concentrated in solutes or lowering the water table, resulting in more rock exposed to air and enhanced mineral oxidation.

SEPN1-related myopathy in three patients: novel mutations and diagnostic clues.

UNLABELLED: Mutations in SEPN1 cause selenoprotein N (SEPN)-related myopathy (SEPN-RM) characterized by early-onset axial and neck weakness, spinal rigidity, respiratory failure and histopathological features, ranging from mild dystrophic signs to a congenital myopathy pattern with myofibrillar disorganization. We report on clinical and instrumental features in three patients affected with a congenital myopathy characterized by prevalent neck weakness starting at different ages and mild myopathy, in whom we performed diagnosis of SEPN-RM. The patients presented myopathic signs since their first years of life, but the disease remained unrecognized because of a relatively benign myopathic course. In two cases, myopathic features were stable after 2 years of follow-up, but respiratory involvement worsened. The muscle MRI and muscle biopsy showed a typical pattern of SEPN-RM. Molecular diagnosis revealed two novel homozygous mutations in SEPN1, c.1176delA and c.726_727InsTCC. CONCLUSION: This report underlines the clinical diagnostic clues of early neck and axial weakness to suspect a SEPN-RM and the usefulness of muscle MRI in conjunction with clinical features to achieve the diagnosis. Our data confirm the slow progression of respiratory involvement in spite of the relatively stable course of myopathy. We report two previously undescribed mutations in SEPN1. WHAT IS KNOWN: • Mutations in SEPN1 cause myopathy characterized by early-onset axial and neck weakness spinal rigidity and respiratory failure. • SEPN-related myopathies have been initially associated with four distinct histopathological entities that however appear more mixed in recently described cases. What is New: • SEPN-related myopathies can remain unrecognized because of the normal early motor development and relatively benign myopathic course of the disease. • Our study adds two novel homozygous mutations to the number of reported pathogenic SEPN1 variants.

Fibrosis and inflammation are greater in muscles of beta-sarcoglycan-null mouse than mdx mouse.

The Sgcb-null mouse, with knocked-down ß-sarcoglycan, develops severe muscular dystrophy as in type 2E human limb girdle muscular dystrophy. The mdx mouse, lacking dystrophin, is the most used model for Duchenne muscular dystrophy (DMD). Unlike DMD, the mdx mouse has mild clinical features and shows little fibrosis in limb muscles. To characterize ECM protein deposition and the progression of muscle fibrosis, we evaluated protein and transcript levels of collagens I, III and VI, decorin, and TGF-ß1, in quadriceps and diaphragm, at 2, 4, 8, 12, 26, and 52 weeks in Sgcb-null mice, and protein levels at 12, 26, and 52 weeks in mdx mice. In Sgcb-null mice, severe morphological disruption was present from 4 weeks in both quadriceps and diaphragm, and included conspicuous deposition of extracellular matrix components. Histopathological features of Sgcb-null mouse muscles were similar to those of age-matched mdx muscles at all ages examined, but, in the Sgcb-null mouse, the extent of connective tissue deposition was generally greater than mdx. Furthermore, in the Sgcb-null mouse, the amount of all three collagen isoforms increased steadily, while, in the mdx, they remained stable. We also found that, at 12 weeks, macrophages were significantly more numerous in mildly inflamed areas of Sgcb-null quadriceps compared to mdx quadriceps (but not in highly inflamed regions), while, in the diaphragm, macrophages did not differ significantly between the two models, in either region. Osteopontin mRNA was also significantly greater at 12 weeks in laser-dissected highly inflamed areas of the Sgcb-null quadriceps compared to the mdx quadriceps. TGF-ß1 was present in areas of degeneration-regeneration, but levels were highly variable and in general did not differ significantly between the two models and controls. The roles of the various subtypes of macrophages in muscle repair and fibrosis in the two models require further study. The Sgcb-null mouse, which develops early fibrosis in limb muscles, appears more promising than the mdx mouse for probing pathogenetic mechanisms of muscle fibrosis and for developing anti-fibrotic treatments. Highlights • The Sgcb-null mouse develops severe muscular dystrophy, the mdx mouse does not. • Fibrosis developed earlier in Sgcb-null quadriceps and diaphragm than mdx. • Macrophages were commoner in mildly inflamed parts of Sgcb-null quadriceps than mdx. • The Sgcb-null model appears more useful than mdx for studying fibrotic mechanisms. • The Sgcb-null model also appears more useful for developing anti-fibrotic treatments.

Total phosphorus reference condition for subalpine lakes: a comparison among traditional methods and a new process-based watershed approach.

Different methods for estimating the total phosphorus (TP) reference conditions of lakes have rarely been compared. This work tests the uncertainty and accuracy of the most frequently used approaches (Morpho-edaphic index -MEI-, export coefficient, diatoms and pigment-inferred TP models) for 35 subalpine lakes. Furthermore, we propose a new process-based watershed approach that was tested on a subalpine environment and consists of combining a space for time substitution with a space for space substitution. The possible presence of uncontaminated or less contaminated environments inside or next to the watershed can be exploited by training a hydrological transport watershed model according to the uncontaminated conditions and then applying the calibration to the entire watershed, which reconstructs a natural or semi-natural TP load scenario. We found that the root mean square error (RMSE) for the MEI is 4 µg L(-1). However, its application is limited for lakes that present with an alkalinity ≤1 meq L(-1). For lakes with a higher alkalinity, we observed a loss of predictive capability that results from the lower solubility of phosphorus under conditions of high calcium content. The export coefficient model was applied with a mean export coefficient and presents similar prediction capabilities as the MEI. The chlorophyll-inferred TP model shows a higher uncertainty (RMSE = 8 µg L(-1)); however, it produced fewer underestimations and overestimations. With regards to the diatom-inferred TP model, we are only able to evaluate an uncertainty of 5 µg L(-1) at the European level. Finally, the proposed process-based watershed approach adequately predicted the reference condition of the selected lake and had an uncertainty lower than the other methods (2 µg L(-1)). We conclude by revealing the potential and limitations of this approach in the field of ecological lake modelling more and more attracted by TP pristine load inputs in studies on the effects of climate change and eutrophication of lakes.

Factors controlling the water quality of rock glacier springs in European and American mountain ranges.

Rock glaciers (RGs) provide significant water resources in mountain areas under climate change. Recent research has highlighted high concentrations of solutes including trace elements in RG-fed waters, with negative implications on water quality. Yet, sparse studies from a few locations hinder conclusions about the main drivers of solute export from RGs. Here, in an unprecedented effort, we collected published and unpublished data on rock glacier hydrochemistry around the globe. We considered 201 RG springs from mountain ranges across Europe, North and South America, using a combination of machine learning, multivariate and univariate analyses, and geochemical modeling. We found that 35 % of springs issuing from intact RGs (containing internal ice) have water quality below drinking water standards, compared to 5 % of springs connected to relict RGs (without internal ice). The interaction of ice and bedrock lithology is responsible for solute concentrations in RG springs. Indeed, we found higher concentrations of sulfate and trace elements in springs sourcing from intact RGs compared to water originating from relict RGs, mostly in specific lithological settings. Enhanced sulfide oxidation in intact RGs is responsible for the elevated trace element concentrations. Challenges for water management may arise in mountain catchments rich in intact RGs, and where the predisposing geology would make these areas geochemical RG hotspots. Our work represents a first comprehensive attempt to identify the main drivers of solute concentrations in RG waters.

Familial adult-onset Pompe disease associated with unusual clinical and histological features.

The adult-onset form of Pompe disease had a wide clinical spectrum, ranging from asymptomatic patients with increased CK to muscle cramps and pain syndrome or rigid-spine syndrome. In addition clinical severity and disease progression are greatly variable. We report on a family with 3 siblings characterized by an unusual adult-onset Pompe disease including dysphagia and weakness of tongue, axial and limb-girdle muscles, in association with atypical globular inclusions in muscle fibres. Our study confirms the great clinical and histological variability of adult-onset Pompe disease and further supports the need of careful evaluation of bulbar function in patients affected by this pathology.

Multiple Carrying Capacities from a management-oriented perspective to operationalize sustainable tourism in protected areas.

This article describes how the concept of Tourism Carrying Capacity (TCC) has shifted from a uni-dimensional approach to incorporating environmental, social and political aspects. This shift is demonstrated by a study of a large, internationally popular protected area used by trekkers, the Mt. Everest Region, where qualitative data collected from visitors was combined with environmental modeling using a participatory framework. Tourist satisfaction showed positive margins for further tourist industry expansion, but current environmental conditions limit growth and further development. Space and time dimensions were also considered. We observed that the limits on growth and further development can be manipulated, with a certain degree of flexibility, through investments and regulatory measures. We hypothesized that TCC can play an important role in the management of protected areas only if it is viewed as a systematic, strategic policy tool within a planning process rather than as a unique, intrinsic number that is not modifiable. We conclude that to translate the strategy into action using standard measures, further investigation is needed to balance the various TCC components as a part of a decision-making framework that includes the integration of different cultural approaches and policy needs.

Suspect screening of wastewaters to trace anti-COVID-19 drugs: Potential adverse effects on aquatic environment.

During the first period of the SARS-CoV-2 pandemic, the lack of specific therapeutic treatments led to the provisional use of a number of drugs, with a continuous review of health protocols when new scientific evidence emerged. The management of this emergency sanitary situation could not take care of the possible indirect adverse effects on the environment, such as the release of a large amount of pharmaceuticals from wastewater treatment plants. The massive use of drugs, which were never used so widely until then, implied new risks for the aquatic environment. In this study, a suspect screening approach using Liquid Chromatography-High Resolution Mass Spectrometry techniques, allowed us to survey the presence of pharmaceuticals used for COVID-19 treatment in three WWTPs of Lombardy region, where the first European cluster of SARS-CoV-2 cases was detected. Starting from a list of sixty-three suspect compounds used against COVID-19 (including some metabolites and transformation products), six compounds were fully identified and monitored together with other target analytes, mainly pharmaceuticals of common use. A monthly monitoring campaign was conducted in a WWTP from April to December 2020 and the temporal trends of some anti-COVID-19 drugs were positively correlated with those of COVID-19 cases and deaths. The comparison of the average emission loads among the three WWTPs evidenced that the highest loads of hydroxychloroquine, azithromycin and ciprofloxacin were measured in the WWTP which received the sewages from a hospital specializing in the treatment of COVID-19 patients. The monitoring of the receiving water bodies evidenced the presence of eight compounds of high ecological concern, whose risk was assessed in terms of toxicity and the possibility of inducing antibiotic and viral resistance. The results clearly showed that the enhanced, but not completely justified, use of ciprofloxacin and azithromycin represented a risk for antibiotic resistance in the aquatic ecosystems.

Twenty-year sediment contamination trends in some tributaries of Lake Maggiore (Northern Italy): relation with anthropogenic factors.

Lake tributaries collect contaminants from the watershed, which may accumulate in lake sediments over time and may be removed through the outlets. DDx, PCB, PAH, PBDE, and trace element (Hg, As, Cd, Ni, Cu, Pb) contamination was analyzed over 2001-2018 period in sediments of the 5 main tributaries and of the outlet of Lake Maggiore (Northern Italy). Sediment cores were collected in two points of the lake, covering 1995-2017 period. Concentrations were compared to Sediment Quality Guidelines (PECs), potential sources and drivers (land use, population numbers, industrial activities, hydrology) were analyzed, and temporal trends were calculated (Mann-Kendall test). PCB, PBDE, Pb, Cd, and Hg contamination derives mainly from heavy urbanization and industry. Cu and Pb show a temporal decreasing trend in the basin, likely as result of improved wastewater treatments and change in use. A recent PAH increase in the whole lake may derive from a single point source. A legacy DDx and Hg industrial pollution is still present, due to high persistence in sediments. Values of DDx, Hg, Pb, and Cu above the PECs in lake sediments and/or in the outlet show potential risk for aquatic organisms. Results highlight the key role of tributaries in driving contamination from the watershed to the lake through sediment transport.

Manganese-mediated hydrochemistry and microbiology in a meromictic subalpine lake (Lake Idro, Northern Italy) - A biogeochemical approach.

This study presents the findings from several field campaigns carried out in Lake Idro (Northern Italy), a deep (124 m) meromictic-subalpine lake, whose water column is subdivided in a mixolimnion (~0-40 m) and a monimolimnion (~40-124 m). Hydrochemical data highlight two main peculiarities characterizing the Lake Idro meromixis: a) presence of a high manganese/iron ratio (up to 20 mol/mol), b) absence of a clear chemocline between the two main layers. The high manganese content contributed to the formation of a stable manganese dominated deep turbid stratum (40-65 m), enveloping the redoxcline (~45-55 m) in the upper monimolimnion. The presence of this turbid stratum in Lake Idro is described for the first time in this study. The paper examines the distribution of dissolved and particulate forms of transition metals (Mn and Fe), alkaline earth metals (Ca and Mg), and other macro-constituents or nutrients (S, P, NO3-N, NH4-N), discussing their behavior over the redoxcline, where the main transition processes occur. Field measurements and theoretical considerations suggest that the deep turbid stratum is formed by a complex mixture of manganese and iron compounds with a prevalence of Mn(II)/Mn(III) in different forms including dissolved, colloidal, and fine particles, that give to the turbid stratum a white-pink opalescent coloration. The bacteria populations show a clear stratification with the upper aerobic layer dominated by the heterotrophic Flavobacterium sp., the turbid stratum hosting a specific microbiological pool, dominated by Caldimonas sp., and the deeper anaerobic layer dominated by the sulfur-oxidizing and denitrifier Sulfuricurvum sp. The occurrence in August 2010 of an anomalous lake surface coloration lasting about four weeks and developing from milky white-green to red-brown suggests that the upper zone of the turbid stratum could be eroded during intense weather-hydrological conditions with the final red-brown coloration resulting from the oxidation of Mn(II)/Mn(III) to Mn(IV) compounds.

Glycogen storage in a zebrafish Pompe disease model is reduced by 3-BrPA treatment.

Pompe disease (PD) is an autosomal recessive muscular disorder caused by deficiency of the glycogen hydrolytic enzyme acid α-glucosidase (GAA). The enzyme replacement therapy, currently the only available therapy for PD patients, is efficacious in improving cardiomyopathy in the infantile form, but not equally effective in the late onset cases with involvement of skeletal muscle. Correction of the skeletal muscle phenotype has indeed been challenging, probably due to concomitant dysfunctional autophagy. The increasing attention to the pathogenic mechanisms of PD and the search of new therapeutic strategies prompted us to generate and characterize a novel transient PD model, using zebrafish. Our model presented increased glycogen content, markedly altered motor behavior and increased lysosome content, in addition to altered expression of the autophagy-related transcripts and proteins Beclin1, p62 and Lc3b. Furthermore, the model was used to assess the beneficial effects of 3-bromopyruvic acid (3-BrPA). Treatment with 3-BrPA induced amelioration of the model phenotypes regarding glycogen storage, motility behavior and autophagy-related transcripts and proteins. Our zebrafish PD model recapitulates most of the defects observed in human patients, proving to be a powerful translational model. Moreover, 3-BrPA unveiled to be a promising compound for treatment of conditions with glycogen accumulation.

Presence and infectivity of SARS-CoV-2 virus in wastewaters and rivers.

The presence of SARS-CoV-2 in raw wastewaters has been demonstrated in many countries affected by this pandemic. Nevertheless, virus presence and infectivity in treated wastewaters, but also in the receiving water bodies are still poorly investigated. In this study, raw and treated samples from three wastewater treatment plants, and three river samples within the Milano Metropolitan Area, Italy, were surveyed for SARS-CoV-2 RNA detection by means of real time RT-PCR and infectivity test on culture cells. SARS-CoV-2 RNA was detected in raw, but not in treated wastewaters (four and two samples, respectively, sampled in two dates). The isolated virus genome was sequenced, and belonged to the strain most spread in Europe and similar to another found in the same region. RNA presence in raw wastewater samples decreased after eight days, probably following the epidemiological trend estimated for the area. Virus infectivity was always null, indicating the natural decay of viral pathogenicity in time from emission. Samples from receiving rivers (three sites, sampled in the same dates as wastewaters) showed in some cases a positivity to real time RT-PCR, probably due to non-treated, or inefficiently treated discharges, or to the combined sewage overflows. Nevertheless, also for rivers infectivity was null. Risks for public health should be limited, although a precautionary approach to risk assessment is here advocated, giving the preliminary nature of the presented data.

Phosphorus content in a deep river sediment core as a tracer of long-term (1962-2011) anthropogenic impacts: A lesson from the Milan metropolitan area.

Reconstructions of past fluvial contamination through the analysis of deep sediment cores are rarely reported in literature. We examined the phosphorus fractions in a deep (2.6 m) sediment core of the Lambro River downstream of the highly anthropized Milan metropolitan area and upstream of the Po river the main Italian watercourse. The core covered the period 1962-2011. Total phosphorus concentrations resulted typical of a strongly impacted environment (4788 mg P kg DW-1 on average) with the highest concentrations related to the 1960s (7639 mg P kg DW-1) reflecting the period of maximum demographic growth. Afterwards, phosphorus concentrations decreased thanks to the infrastructural and legislative initiatives carried out in the 1980s and the 1990s to reduce the impact of urban point sources. Subsequently, total phosphorus concentrations stabilized on values around 3000 mg P kg DW-1 and did not diminish further, even after the second phase of infrastructural interventions carried out in the second half of the 2000s. This was related to the increasing relative impact of the combined sewer overflows in the sewage system and to the strong phosphorus enrichment of the basin. Most of the phosphorus was in inorganic forms (86% of the total) that have been identified as the final target of the domestic effluent inputs. The contribution of organic phosphorus was lower but constant over the period 1962-2011. It likely originated from the agricultural areas located south of the city of Milan. In conclusion, this study underlines how past interventions have been effective in reducing urban point sources but it also highlights the current difficulties related to the growing importance of other sources influenced by the surface runoff (i.e., combined sewer overflows and agriculture). The study also emphasizes a general phosphorus enrichment of the Lambro River basin and its impact on the Po River and the Adriatic Sea.

Urbanization and climate change impacts on surface water quality: Enhancing the resilience by reducing impervious surfaces.

Climate change and urbanization are key factors affecting the future of water quality in urbanized catchments. The work reported in this paper is an evaluation of the combined and relative impact of climate change and urbanization on the water quality of receiving water bodies in the context of a highly urbanized watershed served by a combined sewer system (CSS) in northern Italy. The impact is determined by an integrated modelling study involving two years of field campaigns. The results obtained from the case study show that impervious urban surfaces and rainfall intensity are significant predictors of combined sewer overflows (CSOs) and consequently of the water quality of the receiving water body. Scenarios for the year 2100 demonstrate that climate change combined with increasing urbanization is likely to lead to severe worsening of river water quality due to a doubling of the total phosphorus load from CSOs compared to the current load. Reduction in imperviousness was found to be a suitable strategy to adapt to these scenarios by limiting the construction of new impervious areas and decreasing the existing areas by only 15%. This information can be further utilized to develop future designs, which in turn should make these systems more resilient to future changes in climate and urbanization.

Rainfall as primary driver of discharge and solute export from rock glaciers: The Col d'Olen Rock Glacier in the NW Italian Alps.

Three hypotheses exist to explain how meteorological variables drive the amount and concentration of solute-enriched water from rock glaciers: (1) Warm periods cause increased subsurface ice melt, which releases solutes; (2) rain periods and the melt of long-lasting snow enhance dilution of rock-glacier outflows; and (3) percolation of rain through rock glaciers facilitates the export of solutes, causing an opposite effect as that described in hypothesis (2). This lack of detailed understanding likely exists because suitable studies of meteorological variables, hydrologic processes and chemical characteristics of water bodies downstream from rock glaciers are unavailable. In this study, a rock-glacier pond in the North-Western Italian Alps was studied on a weekly basis for the ice-free seasons 2014 and 2015 by observing the meteorological variables (air temperature, snowmelt, rainfall) assumed to drive the export of solute-enriched waters from the rock glacier and the hydrochemical response of the pond (water temperature as a proxy of rock-glacier discharge, stable water isotopes, major ions and selected trace elements). An intra-seasonal pattern of increasing solute export associated with higher rock-glacier discharge was found. Specifically, rainfall, after the winter snowpack depletion and prolonged periods of atmospheric temperature above 0 °C, was found to be the primary driver of solute export from the rock glacier during the ice-free season. This occurs likely through the flushing of isotopically- and geochemically-enriched icemelt, causing concomitant increases in the rock-glacier discharge and the solute export (SO42-, Mg2+, Ca2+, Ni, Mn, Co). Moreover, flushing of microbially-active sediments can cause increases in NO3- export.

Up-regulation of Toll-like receptors 7 and 9 and its potential implications in the pathogenic mechanisms of LMNA-related myopathies.

Laminopathies are a heterogeneous group of diseases, caused by mutations in lamin A/C proteins. The most common laminopathy (LMNA-related myopathies, LMNA-RM) affects skeletal and cardiac muscles; muscle histopathology is variable, ranging from mild unspecific changes to dystrophic features, sometimes with inflammatory evidence. Whether the genetic defect might activate innate immune components, leading to chronic inflammation, myofiber necrosis and fibrosis, is still unknown. By qPCR, a significant up-regulation of Toll-like receptor (TLR) 7 and 9 transcripts was found in LMNA-RM compared to other myopathic and non-myopathic muscles. A marked TLR7/9 staining was observed on LMNA-RM blood vessels and muscle fibers and, when present, on infiltrating cells, mainly macrophages, scattered in the tissue or localized close to degenerated muscle fibers and connective tissue. Our results recognize innate immunity as a player in LMNA-RM pathogenesis. Modulation of TLR7/9 signaling pathways and decrease of macrophage-mediated inflammation might be potential therapeutic strategies in LMNA-RM management. ABBREVIATIONS: DMD, Duchenne muscular dystrophy; EDMD2, Emery-Dreifuss muscular dystrophy type 2; FSHD, facio-scapulo-humeral muscular dystrophy; LGMD1B, limb-girdle muscular dystrophy type 1B; LMNA-CMD, LMNA-related congenital muscular dystrophy; LMNA-RM, LMNA-related myopathies; sIBM, sporadic inclusion body myositis; TLR, Toll-like receptor.