ALBERT

Description: Abstract We integrate paleoseismic datasets along the Mt. Vettore‐Mt. Bove normal fault‐system (VBFS) rupturing at surface in the 30 October 2016 Norcia earthquake. Through the analysis of new trenches from this work and a review of the pre‐existing data, we correlate events among trench sites along antithetic and synthetic fault splays. We recognize seven M6.5, 2016 Norcia‐type (or larger) surface‐faulting events in the last ~22 kyr, including 2016. Before 2016, one event occurred in the past two millennia (260‐575 CE), and possibly corresponds to the event damaging Rome in 443 CE or 484/508 CE. Three previous events occurred between 10590 BCE and 415 BCE, whereas the two oldest ones date between 19820 BCE and 16540 BCE. The average recurrence time is 3360–3640 yrs for the last ~22 kyr, and 1220‐1970 yrs for the last ~4 kyr. We infer a minimum dip‐slip rate of 0.26‐0.38 mm/yr on the master fault in the central portion of the VBFS, and a dip‐slip rate of at least 0.10 mm/yr on the southernmost portion. We infer a Middle‐Late Pleistocene inception of the long‐term scarp of the investigated splays. The along‐strike variation of slip rates well reproduces the trend of the 2016 surface slip, thus the time window exposed in the trenches is representative for the present fault activity. Based on trenching data, different earthquake rupture scenarios should be also considered for local hazard assessment.

Description: An artificial wound in a confluent monolayer of human keratinocyte HaCaT cells or mouse embryo fibroblast Swiss NIH 3T3 cells was used to analyze the effects of the nitric oxide (NO) chemical donor, S-nitroso-N-acetylpenicillamine (SNAP). SNAP exposure promoted an enhanced rate of wound closure and accelerated motility of both keratinocytes and fibroblasts compared to control cells. The wounded monolayer cultures of HaCaT and NIH 3T3 cells, treated with or without SNAP, were monitored under a phase contrast microscope. Structural and ultrastructural modifications were analysed by scanning electron microscopy (SEM). The images were captured by a digital camera at different time points (0-28 h) and the wound area was analysed through software included in Matlab®. As early as 15 minutes, SNAP induced significant cytoskeletal remodelling, as shown by immunostaining (phalloidin-labelling), which in turn was associated with increased filopodium number and length rise. NO donor treatment also induced overexpression of Ki-67 protein, a typical marker of cell proliferation, as shown by immunostaining. Both SNAP-induced migration and proliferation were antagonized by the NO-sensitive GC inhibitor 1H-[1,2,4]oxadiazolo[-4,3-a]quinoxalin-1-one (ODQ), which suggests activation of the NO/cGMP signalling cascade in the observed SNAP-induced effects in the early stages of the healing process. Moreover, we provide evidence that PPAR-beta antagonist (GSK0660) may interfere with NO-mediated wound healing process. This article is protected by copyright. All rights reserved

Description: Complex scattering problems are often made by composite structures where wedges and penetrable substrates may interact at near field. In this paper (Part I) together with its companion paper (Part II) we study the canonical problem constituted of a Perfectly Electrically Conducting (PEC) wedge lying on a grounded dielectric slab with a comprehensive mathematical model based on the application of the Generalized Wiener-Hopf Technique (GWHT) with the help of equivalent circuital representations for linear homogenous regions (angular and layered region). The proposed procedure is valid for the general case and the papers focus on E-polarization. The solution is obtained using analytical and semi-analytical approaches that reduce the Wiener-Hopf factorization to integral equations. Several numerical test cases validate the proposed method. The scope of Part I is to present the method and its validation applied to the problem. The companion paper Part II focuses on the properties of the solution and it presents physical and engineering insights as GTD/UTD diffraction coefficients, total far fields, modal fields and excitation of surface and leaky waves for different kinds of source. The structure is of interest in antenna technologies and electromagnetic compatibility (tip on a substrate with guiding and antenna properties).

Description: Together with Part I, Part II describes the theory, the validation and the application of the Generalized Wiener Hopf Technique (GWHT) to complex scattering problems constituted of wedges and layers that may interact at near field. In particular we present the full analysis of the canonical problem where a Perfectly Electrically Conducting (PEC) wedge is lying on a grounded dielectric slab bounded by a PEC plane. The structure proposed in the two papers is of interest in antenna technologies and electromagnetic compatibility (tip on a substrate with guiding and antenna properties). While the scope of Part I has been to present the method and its validation applied to the problem, this paper (Part II) focuses on the properties of the solution and it presents physical and engineering insights. Part II starts from the solution of Generalized Wiener Hopf Equations (GWHEs) through Fredholm factorization in terms of an analytical element of the spectral unknowns (Part I) and it illustrates how to estimate physical and engineering characteristics of the problem: GTD/UTD diffraction coefficients, total far fields, modal fields and excitation of surface waves and leaky waves for different excitations (modal source and/or plane wave source).

Description: Tree mortality is a key factor influencing forest functions and dynamics, but our understanding of the mechanisms leading to mortality and the associated changes in tree growth rates are still limited. We compiled a new pan-continental tree-ring width database from sites where both dead and living trees were sampled (2,970 dead and 4,224 living trees from 190 sites, including 36 species), and compared early and recent growth rates between trees that died and those that survived a given mortality event. We observed a decrease in radial growth before death in ca. 84% of the mortality events. The extent and duration of these reductions were highly variable (1-100 years in 96% of events) due to the complex interactions among study species and the source(s) of mortality. Strong and long-lasting declines were found for gymnosperms, shade- and drought-tolerant species, and trees that died from competition. Angiosperms and trees that died due to biotic attacks (especially bark-beetles) typically showed relatively small and short-term growth reductions. Our analysis did not highlight any universal trade-off between early growth and tree longevity within a species, although this result may also reflect high variability in sampling design among sites. The inter-site and inter-specific variability in growth patterns before mortality provides valuable information on the nature of the mortality process, which is consistent with our understanding of the physiological mechanisms leading to mortality. Abrupt changes in growth immediately before death can be associated with generalized hydraulic failure and/or bark beetle attack, while long-term decrease in growth may be associated with a gradual decline in hydraulic performance coupled with depletion in carbon reserves. Our results imply that growth-based mortality algorithms may be a powerful tool for predicting gymnosperm mortality induced by chronic stress, but not necessarily so for angiosperms and in case of intense drought or bark-beetle outbreaks. This article is protected by copyright. All rights reserved.

Description: Maintaining multiple ecological functions (‘multifunctionality’) is crucial to sustain viable ecosystems. To date most studies on biodiversity-ecosystem functioning (BEF) have focused on single or few ecological functions and services. However, there is a critical need to evaluate how species and species assemblages affect multiple processes at the same time, and how these functions are interconnected. Dung beetles represent excellent model organisms because they are key contributors to several ecosystem functions. Using a novel method based on the application of 15 N-enriched dung in a mesocosm field experiment, we assessed the role of dung beetles in regulating multiple aspects of nutrient cycling in alpine pastures over appropriate spatial (up to a soil depth of 20 cm) and temporal (up to one year after dung application) scales. 15 N isotope tracing allowed the evaluation of multiple interrelated ecosystem functions responsible for the cycling of dung-derived nitrogen (DDN) in the soil and vegetation. We also resolved the role of functional group identity and the importance of interactions among co-occurring species for sustaining multiple functions by focusing on two different dung beetle nesting strategies (tunnelers and dwellers). Species interactions were studied by contrasting mixed-species to single-species assemblages, and asking whether the former performed multiple functions better than the latter. Dung beetles influenced at least seven ecological functions by facilitating dung removal, transport of DDN into the soil, microbial ammonification and nitrification processes, uptake of DDN by plants, herbage growth, and changes in botanical composition. Tunnelers and dwellers were found to be similarly efficient for most functions, with differences based on the spatial and temporal scales over which the functions operated. Although mixed-species assemblages seemed to perform better than single-species, this outcome may be dependent on the context. Most importantly though, the different functions were found to be interconnected sequentially as reveled by analyzing 15 N content in dung, soil and vegetation. Taken together, our current findings offer strong support for the contention that the link between biodiversity and ecosystem functions should be examined not function by function, but in terms of understanding multiple functions and how they interact with each other. This article is protected by copyright. All rights reserved.

Description: ABSTRACT Autosomal recessive Pompe disease is a lysosomal disorder caused by mutations of the acid-α-glucosidase (GAA) gene. Deficiency of GAA enzyme leads to glycogen accumulation and autophagy impairment in cardiac and skeletal muscles, but also in lymphocytes. Since an effective therapy is available, a rapid, sensitive and specific test is crucial to early identify affected subjects. Number of lymphocytes containing PAS-positive vacuoles was evaluated on blood films from 72 consecutive adult patients with hyperckemia and/or muscle weakness, 13 genetically confirmed late-onset-Pompe-disease (LOPD) and 13 of their offspring. GAA activity, measured on dried blood spot (DBS) in all patients inversely correlated with number of PAS-positive lymphocytes. More than 4 PAS-positive lymphocytes were found in 11 out of the 72 patients (6 new diagnosis of LOPD, 3 different glycogen storage myopathies, 1 glucose-6-phosphate dehydrogenase deficiency, 1 caveolinopathy), in all 13 LOPD patients and in the 13 LOPD offspring. These latter resulted to have all a single GAA mutation but low GAA levels. Immunostaining with the autophagy markers LC3 and p62 confirmed the autophagic nature of lymphocytes vacuoles. ROC curve assessment of PAS-positive lymphocytes disclosed 100% of sensitivity and 94% of specificity in recognizing both compound heterozygous and heterozygous GAA carriers. The other myopathies with more than 4 PAS-positive lymphocytes appeared to be all related to impaired autophagy, which seems to be responsible of PAS-positive vacuolated lymphocytes formation. Quantification of PAS-positive lymphocytes in blood films is useful to identify autophagic vacuolar myopathies and should be routinely used as first level test for Pompe disease. This article is protected by copyright. All rights reserved

Description: Quantifying root biomass in rocky forest soils is challenging. This report provides practical advice for field sampling and laboratory processing of root biomass in these settings. Manual coring is the most efficient method for sampling fine root biomass in the upper soil profile (we sampled to 30 cm). However, careful correction for coarse fragment volume is needed because manual coring is impeded by rocks. Unbiased estimation of root biomass below obstructions requires either excavating a pit or power coring. We recommend power coring because of the very high field labor costs of pit excavation. Roots can be separated from soil either by dry picking or by wet sieving. For surface organic matter-rich horizons typical of many forest soils, only dry picking is feasible. A timed interval approach can greatly reduce laboratory processing time. Because sorting live from dead roots is necessarily subjective, efforts to avoid fragmentation of root systems obtained from cores are strongly recommended. Sample size requirements for detecting changes or differences in root biomass at the stand level are presented based on extensive sampling in northern hardwood forests. Detecting 20% differences in fine root (〈1 mm) biomass in 0–30 cm soil using ten 5-cm manual cores generally would require about nine sample plots in a stand, whereas detecting such differences in deep soil (30–50 cm) would be virtually impossible because of extreme spatial variation. Power analyses such as these can help improve experimental designs, as the spatial intensity of sampling determines the detectable difference, which can in turn guide decisions about the temporal frequency of sampling.

Description: The data here reported introduce the wound-healing assay as a tool for testing probiotics aimed at protecting gastrointestinal mucosal surfaces and to verify the consistency of their manufacturing. At the scope, we compared the in vitro effects of two multi-strain high concentration formulations both commercialized under the same brand VSL#3 but sourced from different production sites (USA and Italy) on a non-transformed small-intestinal epithelial cell line, IEC-6. The effects on cellular morphology, viability, migration, and H 2 O 2 -induced damage, were assessed before and after the treatment with both VSL#3 formulations. While the USA-sourced product (“USA-made”) VSL#3 did not affect monolayer morphology and cellular density, the addition of bacteria from the Italy-derived product (“Italy-made”) VSL#3 caused clear morphological cell damage and strongly reduced cellularity. The treatment with “USA-made” lysate led to a higher rate of wounded monolayer healing, while the addition of “Italy-made” bacterial lysate did not influence the closure rate as compared to untreated cells. While lysates from “USA-made” VSL#3 clearly enhanced the formation of elongated and aligned stress fibers, “Italy-made” lysates had not similar effect. “USA-made” lysate was able to cause a total inhibition of H 2 O 2 -induced cytotoxic effect whereas “Italy-made” VSL#3 lysate was unable to protect IEC-6 cells from H 2 O 2 -induced damage. ROS generation was also differently influenced, thus supporting the hypotesis of a protective action of “USA-made” VSL#3 lysates, as well as the idea that “Italy-made” formulation was unable to prevent significantly the H 2 O 2 -induced oxidative stress. This article is protected by copyright. All rights reserved

Description: Nowadays, fat tissue transplantation is widely used in regenerative and reconstructive surgery. However, a shared method of lipoaspirate handling for ensuring a good quality fat transplant has not yet been established. The study was to identify a method to recover from the lipoaspirate samples the highest number of human viable adipose tissue-derived stem cells (hADSCs) included in stromal vascular fraction (SVF) cells and of adipocytes suitable for transplantation, avoiding an extreme handling. We compared the lipoaspirate spontaneous stratification (10-20-30 min) with the centrifugation technique at different speeds (90-400-1500 xg ). After each procedure, lipoaspirate was separated into top oily lipid layer, liquid fraction, “middle layer” and bottom layer. We assessed the number of both adipocytes in the middle layer and SVF cells in all layers. The histology of middle layer and the surface phenotype of SVF cells by stemness markers (CD105 + , CD90 + , CD45-) were analyzed as well. The results showed a normal architecture in all conditions except for samples centrifuged at 1500 xg . In both methods, the flow cytometry analysis showed that greater number of ADSCs was in middle layer; in the fluid portion and in bottom layer was not revealed significant expression levels of stemness markers. Our findings indicate that spontaneous stratification at 20 min and centrifugation at 400 xg are efficient approaches to obtain highly viable ADSCs cells and adipocytes, ensuring a good thickness of lipoaspirate for autologous fat transfer. Since an important aspect of surgery practice consists of gain time, the 400 xg centrifugation could be the recommended method when the necessary instrumentation is available. This article is protected by copyright. All rights reserved