ALBERT

Description: It is known that bradykinin (BK) B2 receptor (B2R) is expressed in the collecting duct (CD) cells of the newborn rat kidney, but little is known about its role during early postnatal life. Therefore, we hypothesize that BK could participate in the mechanisms that mediate CD formation during the postnatal renal development. Performing primary cultures, combined with biochemical, immunocytochemical, and time-lapse analysis, we studied the role of BK in CD cell behaviour isolated from renal papilla of neonatal rats. A reverse relationship was observed between B2R expression and the degree of CD epithelial cell sheet maturation. BK stimulation induced CD cell association upon B2R activation. The lack of B2R expression in cells showing mature adherens junctions suggested that BK is mostly involved in early adhesive events, thus favouring the initial formation of CD during development. Time-lapse analysis revealed that BK induced a high protrusive activity of CD cells, denoted by ruffle formation and lamellipodia extension. PI3K was involved in the BK-induced CD cell-cell association and the acquisition of the migratory phenotype since, when inhibited, membrane ruffles and filopodia between cells diminished. Results indicate that the actions of BK mediated by PI3K activation were due to the downstream Akt and Rac pathways. This study, performed with CD cells that were not genetically manipulated, provides new experimental evidence supporting a novel role of BK in rat renal CD organization. Since B2R blockade results in abnormal tubular differentiation, our results contribute to better understanding the etiology of human congenital renal malformation and diseases. This article is protected by copyright. All rights reserved

Description: Sustainability, Vol. 10, Pages 3274: Assessing Urban Fragmentation at Regional Scale Using Sprinkling Indexes Sustainability doi: 10.3390/su10093274 Authors: Lucia Saganeiti Antonella Favale Angela Pilogallo Francesco Scorza Beniamino Murgante Artificial land use trends could represent an effective indicator of the settlement process quality and could also provide information about the efficacy of protection and exploitation policies in natural and rural areas. This work discusses an analytic procedure for the time series investigation of urban settlement development at the regional scale to verify the nexus between urban growth and demographic trends connected with the phenomenon of land take. In Italy, since 1950, the land take phenomenon has been a consequence of several factors: urbanization, realization of transport infrastructures including ports, airports, and highways, and the enhancement of industrial and productive systems. We analyzed all these territorial transformations that create waterproof soil, and more generally, a transition from natural and semi-natural uses toward artificial land use. After World War II, the demographic growth and the consequent housing demand generated a strong urbanization process in the main poles of economic development areas in Italy. Since the early 2000s, the situation has completely changed and the land take phenomenon is no longer mainly based on real need for new urban expansion areas based on effective urban planning tools, but is strongly related to a scattered demand for new housing in a weak territorial spatial planning system not able to drive effective urban development that minimizes speculative real estate initiatives. This uncontrolled occupation of soil generated, in Italy, a landscape fragmentation called the urban sprinkling phenomenon, different from urban sprawl, which is a wider phenomenon characterized by disordered urban growth. The present document aims to assess how uncontrolled expansion in areas characterized by low settlement density can generate fragmentation. To define if the territory is affected by the urban sprinkling phenomenon, two 50-year time series concerning urban expansion of buildings and demographic trends are analyzed calculating population and building density indices and their variation over the years. The sprinkling index is used to analyze the variation in the fragmentation degree at two different scales (regional and municipal). Finally, we discuss the context where this phenomenon has developed, analyzing the buildings located in hydrogeological risk zones and protected areas, and the correlation between demographic changes and the degree of territorial fragmentation variation.

Description: Key Points MPL P106L induces thrombocytosis due to an incomplete trafficking defect that allows very low cell-surface levels. The P106L mutation uncouples MPL signaling from its THPO clearance functions.

Description: Introduction: Allogeneic hematopoietic stem cell transplantation (HSCT) is a well-established therapeutic modality for a variety of hematological malignancies. Unfortunately it is associated with significant morbidity and mortality related to cancer relapse and transplant complications, including graft versus host disease (GvHD). GvHD-free, relapse-free survival (GRFS) is a recently reported composite end point which allows estimating risk of death, relapse and GvHD simultaneously [Holtan et al., Blood 2015 ]. T-cell depletion (TCD) is a well established strategy for GvHD prevention, but is probably associated with increased risk of relapse. In the present work we investigated the effect of partial TCD (pTCD) on GRFS in order to evaluate its impact on patientsÕ morbidity-free survival. Patients and methods: We performed a retrospective study on 333 patients who underwent allogeneic HSCT for hematologic malignancies at our center from 2004 to 2014 with grafts from HLA identical siblings or HLA 10/10 matched unrelated donors. 171 patients received pTCD grafts, obtained through incubation with alemtuzumab in vitro washed before infusion followed on day +1 by an add-back of donor T cells. 162 patients received T cell repleted (non-TCD) grafts. Donor lymphocyte infusions were given at three months to all patients without GvHD who had received pTCD grafts with reduced intensity conditioning and when needed to patients, transplanted with either pTCD or non-TCD grafts with mixed chimerism. Kaplan-Meier estimates were employed to determine the probability of 1-year and 5-year overall survival (OS), progression free survival (PFS) and GRFS. Events determining GRFS included grade 3-4 acute GvHD, systemic therapy-requiring chronic GvHD, relapse, or death. Differences between survival curves were determined using Log-rank Mantel-Cox test. Cox regression was used to examine the independent effect on OS, PFS and GRFS of clinical factors including age, underling disease, disease status at transplant, disease risk index, conditioning, donor type, stem cell source, year of transplantation and T-cell depletion. Cumulative incidence estimates of relapse and non-relapse mortality (NRM) were calculated with relapse or death from other causes defined as competitive events with the Fine and Gray method. Results: According to institutional practices, the group receiving pTCD grafts comprised more patients transplanted in complete remission (67%) than the group receiving non-TCD grafts (41%, p

Description: Key Points Enrichment of atypical MPL mutations in essential thrombocythemia. MPLS204P and MPLY591N mutants are weak gain-of-function mutants.

Description: Myeloproliferative neoplasms (MPNs) are clonal malignant disorders characterized by the increased production of mature myeloid cells in blood. The classical MPNs include Polycythemia Vera (PV), Essential Thrombocythemia (ET) and Primary Myelofibrosis (PMF). Those pathologies are due to the acquisition of gain-of-function mutations leading to the constitutive activation of the cytokine receptor / JAK2 signaling pathway: JAK2V617F in 70% of cases and mutations in the thrombopoietin receptor (MPL) gene in 5% of cases. More recently, around fifty different mutations in the calreticulin (CALR) gene have been described in 30% of ET and PMF with two more frequent mutations called del52 (type 1) and ins5 (type 2). All the CALR mutations induce a frameshiflt to an alternative reading frame in the exon 9 leading to a new C-term tail of the protein with hydrophobic features, and the loss of the KDEL sequence, which is involved in its endoplasmic reticulum retention. The goal of this work was to understand the role of CALR mutants (del52, del46, del34, ins5, del19, del13) in human hematopoiesis. By studying the variant allele frequency (VAF) in 20 patients, we have shown that the CALR mutations are present in all blood mature cells not only in granulocytes and monocytes (CD14+) with a VAF 〉30% but also in B cells (CD19+), NK cells (CD56+) and in some cases in T cells (CD3+). Moreover, we have observed that CALR mutations are present in all hematopoietic progenitors including CD34+CD38-CD90+ (HSC), CD34+CD38-CD90- (immature progenitors) and CD34+CD38+ (committed progenitors) cell fractions after investigating the clonal architecture of the progenitors. CALR mutation was detectable in more than 40% of progenitor cells except in 2 patients (15 patients studied) and with, in some cases, no detectable wild type CALR progenitors. Homozygous CALR mutations were rare except in one case associated with disease progression. Whatever the VAF, there was no significant differences among the different progenitor types and granulocytes. Finally, we observed that all the associated mutations studied (TET2, PHF6, SYNE1, SCARA5, PIK3CD, SETD1B) in 6 patients postdated CALR mutations. We could also show in 15 patients samples that CALR mutants give a specific megakaryocytic progenitor (CFU-MK) spontaneous growth mediated both by MPL and JAK2 activation using specific inhibitors and short hairpin RNAs. The CFU-MK spontaneous growth correlated with a constitutive activation of JAK2/STAT3/5 pathway in megakaryocytes derived from in vitro cultures of CD34+ progenitors. In aggregate, these results show that all CALR mutants studied are present in all human hematopoietic cells including myeloid and lymphoid cells, give an early clonal advantage at the level of the HSC compartment and a specific increased growth of the megakaryocytic lineage via MPL/JAK2 activation. Disclosures No relevant conflicts of interest to declare.

Description: The uncontrolled activation of the immune system toward antigens contained in the gut lumen in genetically predisposed subjects is believed to be the leading cause of inflammatory bowel disease (IBD). Two not mutually exclusive hypotheses can explain the pathogenic process leading to IBD. The first and mostly explored hypothesis states that the loss of tolerance toward gut microbiota antigens generates an aberrant inflammatory response that is perpetuated by continuous and unavoidable exposure to the triggering antigens. However, the discovery that the resolution of inflammation is not the mere consequence of clearing inflammatory triggers and diluting pro-inflammatory factors, but rather an active process in which molecular and cellular elements are involved, implies that a defect in the pro-resolving mechanisms might cause chronic inflammation in different immune-mediated diseases, including IBD. Here we review data on pro-resolving and counter-regulatory mechanisms involved in the resolution of inflammation, aiming to identify their possible involvement in the pathogenesis of IBD.

Description: Lead pacemaker infection is a complication on the rise. An infected oscillating mass attached to the leads (ILV) is a common finding in this setting. Percutaneous extraction of the leads and of the device is the best curative option. However, extraction of leads with large masses can be complicated by pulmonary embolism. The aim of this study was to understand the factors associated with large ILV using a sophisticated ultrasound technique to visualize the masses, namely intracardiac echocardiography (ICE), and investigate whether larger masses induce more complications during and after extraction. Percutaneous lead extraction and peri-procedural ICE were done in 36 patients (pts) (75 ± 11 years old, 74% males). Vegetations (max dimension = 8.2 ± 4.1 mm) in the right cavity were found in 26 of them, mostly adhering to the leads. We subdivided the patients into 2 groups: with vegetation size 〈 1 cm (18 pts) and vegetation size ≥ 1 cm (8 pts). By univariate analysis, we found that patients in group 1 were more often taking anticoagulation therapy (p = 0.03, Phi (Phi coefficient) = −0.5, OR [odds ratio] 0.071) and had signs of local pocket infection (p = 0.02, Phi = −0.52, OR 0.059) while significantly more patients in group 2 had diabetes (p = 0.08, Phi = 0.566, OR 15); moreover the patients in group 2 showed a trend toward a more frequent positive blood culture (p = 0.08, Phi = 0.39, OR 5.8) and infection with coagulase negative staphylococci (p = 0.06, Phi = 0.46, OR 8.3). At multivariate analysis, only 3 factors (diabetes, younger age and anticoagulation therapy) were independently associated with ILV size: diabetes, associated with larger vegetations (group 2), showed the largest beta value (0.44, p = 0.008); age was inversely correlated with ILV size (beta value = −32, p = 0.038), and anticoagulation therapy (beta value = −029, p = 0.048) was more commonly associated with smaller vegetations (group 1). Larger ILV were not associated with more complications or death during or after the extraction. Conclusion: diabetes, anticoagulation therapy and age are independent predictors of lead vegetation size. The embolic potential of large ILV during extraction was modest, so ILVs 〉1cm are not a contraindication to percutaneous extraction of infected leads.

Description: Artificial land use trends could represent an effective indicator of the settlement process quality and could also provide information about the efficacy of protection and exploitation policies in natural and rural areas. This work discusses an analytic procedure for the time series investigation of urban settlement development at the regional scale to verify the nexus between urban growth and demographic trends connected with the phenomenon of land take. In Italy, since 1950, the land take phenomenon has been a consequence of several factors: urbanization, realization of transport infrastructures including ports, airports, and highways, and the enhancement of industrial and productive systems. We analyzed all these territorial transformations that create waterproof soil, and more generally, a transition from natural and semi-natural uses toward artificial land use. After World War II, the demographic growth and the consequent housing demand generated a strong urbanization process in the main poles of economic development areas in Italy. Since the early 2000s, the situation has completely changed and the land take phenomenon is no longer mainly based on real need for new urban expansion areas based on effective urban planning tools, but is strongly related to a scattered demand for new housing in a weak territorial spatial planning system not able to drive effective urban development that minimizes speculative real estate initiatives. This uncontrolled occupation of soil generated, in Italy, a landscape fragmentation called the urban sprinkling phenomenon, different from urban sprawl, which is a wider phenomenon characterized by disordered urban growth. The present document aims to assess how uncontrolled expansion in areas characterized by low settlement density can generate fragmentation. To define if the territory is affected by the urban sprinkling phenomenon, two 50-year time series concerning urban expansion of buildings and demographic trends are analyzed calculating population and building density indices and their variation over the years. The sprinkling index is used to analyze the variation in the fragmentation degree at two different scales (regional and municipal). Finally, we discuss the context where this phenomenon has developed, analyzing the buildings located in hydrogeological risk zones and protected areas, and the correlation between demographic changes and the degree of territorial fragmentation variation.