In vitro immunomodulatory effect of NGAL on PBMCs derived from haemodialysis and hyperimmunized patients

Negli ultimi anni è cresciuto l’interesse verso pazienti in attesa di un trapianto di rene identificati per la presenza elevata di alloanticorpi contro antigeni HLA e non-HLA, definiti iperimmuni. Recentemente, è stato dimostrato l’aumento dell'immunotolleranza ad opera della componente linfocitaria T regolatoria (Treg) dopo l'infusione di MSCs in modelli sperimentali di trapianto d’organo. Alcuni studi hanno infatti dimostrato il contributo di Treg nella modulazione della risposta immunitaria in diversi modelli animali e nel trapianto clinico, per suggerire il loro potenziale utilizzo come marcatori di tolleranza, rigetto o previsione dell’outcome del trapianto d'organo. Un precedente studio ha dimostrato che NGAL (Neutrophil gelatinase-associated lipocalin), un noto biomarker di danno renale, è in grado di indurre in vitro immunotolleranza aumentando l'espressione di HLA-G e l'espansione di Treg in cellule linfo-monocitarie (PBMC) di soggetti sani. In questo studio, si è valutato l'effetto di NGAL sull'espansione di Treg e l'espressione di HLA-G in pazienti con insufficienza renale cronica sottoposti ad emodialisi e iperimmuni, e si è approfondita la capacità immunomodulatoria di cellule staminali mesenchimali adulte derivate da tessuto adiposo (ASC) nei confronti di PBMC derivati da queste categorie di pazienti. I nostri risultati hanno dimostrato un effetto immunomodulatorio, dose dipendente, mediato da NGAL sulla popolazione linfo-monocitaria derivata da pazienti in emodialisi non iperimmuni evidenziato dall’aumento della popolazione di Treg e dall'espressione di HLA-G. Questo effetto è risultato inibito in PBMC derivati da pazienti iperimmuni. L'immunomodulazione sui PBMC in quest’ultima categoria di pazienti è stata ristabilita in vitro in seguito all’allestimento di co-colture con cellule staminali mesenchimali derivate da tessuto adiposo. Le ASC risultano una buona prospettiva futura di terapia cellulare autologa, in concomitanza e a supporto del trapianto di rene, in pazienti uno stato immunologico alterato, ampliando l'accesso ai trapianti d’organo a questi pazienti complessi che presentano un PRA sfavorevole.In recent years, an increasing number of patients waiting for a kidney transplant showed the presence of alloantibodies against HLA antigens and non-HLA. Evidence demonstrates the contribution of regulatory T lymphocyte cells (Treg) in modulating the immune response in different animal models and in clinical transplant, to suggest their potential use as markers of tolerance, rejection or prediction of organ transplant outcome. Recently, Treg has been shown to increase immunotolerance following MSCs infusion into experimental models of solid organ transplantation. We have previously provided in vitro evidence that NGAL (Neutrophil gelatinase-associated lipocalin), a known biomarker of renal injury, is able to induce immune tolerance by upregulating HLA-G expression and expansion of Treg in normal subjects. In this study, we evaluated the effect of NGAL on the expansion of Treg and HLA-G expression in haemodialysis and hyperimmunized patients; then, we investigated the immunomodulatory effect of adipose-derived mesenchymal stem cells (ASCs) on PBMCs derived from patients. We carried out isolation from peripheral blood mononuclear cells (PBMCs) and immunophenotypic in vitro characterization of Treg after NGAL treatment and after co-culture with ASCs. Our results demonstrated an immunomodulatory effect NGAL-mediated on lymphomonocyte population derived by non-hyperimmunized, as Treg population and HLA-G expression augmented, while this effect is inhibited in PBMCs derived from hyperimmunized patients. The immunomodulatory effect on PBMCs in these patients is re-established in vitro by co-culture with adipose-derived mesenchymal stem cells. ASCs could be a good future possibility for modulating immune system before or during kidney transplant in hyperimmunized patients, expanding access to transplants to these complex patients

A wireless method to obtain the impedance from scattering parameters

The coaxial wire method is a common and appreciated choice to assess the beam coupling impedance (BCI) of an accelerator element. Nevertheless, the results obtained from wire measurements could be inaccurate due to the presence of the stretched conductive. The aim of this work is to establish a solid technique to obtain the BCI from electromagnetic simulations, without modifications of the device under test. In this framework, we identified a new relation to get the resistive wall beam coupling impedance of a circular chamber directly from the scattering parameters. Furthermore, a possible generalization of the method to arbitrary cross section geometries has been studied and validated with numerical simulations

Unveiling the morphogenetic code: A new path at the intersection of physical energies and chemical signaling

In this editorial, we discuss the remarkable role of physical energies in the control of cell signaling networks and in the specification of the architectural plan of both somatic and stem cells. In particular, we focus on the biological relevance of bioelectricity in the pattern control that orchestrates both developmental and regenerative pathways. To this end, the narrative starts from the dawn of the first studies on animal electricity, reconsidering the pioneer work of Harold Saxton Burr in the light of the current achievements. We finally discuss the most recent evidence showing that bioelectric signaling is an essential component of the informational processes that control pattern specification during embryogenesis, regeneration, or even malignant transformation. We conclude that there is now mounting evidence for the existence of a Morphogenetic Code, and that deciphering this code may lead to unprecedented opportunities for the development of novel paradigms of cure in regenerative and precision medicine

Development of a redox polymer based on poly(2-hydroxyethyl methacrylate)for disposable amperometric sensors

Membranes based on poly(2-hydroxyethyl methacrylate), chemically modified with ferrocene functionalities, are proposed as novel coatings for the development of disposable amperometric sensors. They are mass-produced on an inert support through an UV photo-induced polymerization, forming self-standing films that can be transferred on an electrode surface. Thanks to the characteristic of the functionalized methacrylate matrix to rapidly and reversibly swell by incorporation of large amounts of water, the ferrocene moieties are in intimate contact with the electrolytic solution. They can activate effective electrocatalytic processes that can be exploited in the field of amperometric sensing. The performance of the methacrylate coating with respect to the determination of hydroquinone benchmark analyte has been tested both in a static solution and in a flux cell

Quality Control Platform for the Standardization of a Regenerative Medicine Product

Adipose tissue is an attractive source of stem cells due to its wide availability. They contribute to the stromal vascular fraction (SVF), which is composed of pre-adipocytes, tissue-progenitors, and pericytes, among others. Because its direct use in medical applications is increasing worldwide, new quality control systems are required. We investigated the ability of the Non-Equilibrium Earth Gravity Assisted Dynamic Fractionation (NEEGA-DF) method to analyze and separate cells based solely on their physical characteristics, resulting in a fingerprint of the biological sample. Adipose tissue was enzymatically digested, and the SVF was analyzed by NEEGA-DF. Based on the fractogram (the UV signal of eluting cells versus time of analysis) the collection time was set to sort alive cells. The collected cells (F-SVF) were analyzed for their phenotype, immunomodulation ability, and differentiation potential. The SVF profile showed reproducibility, and the alive cells were collected. The F-SVF showed intact adhesion phenotype, proliferation, and differentiation potential. The methodology allowed enrichment of the mesenchymal component with a higher expression of mesenchymal markers and depletion of debris, RBCs, and an extracellular matrix still present in the digestive product. Moreover, cells eluting in the last minutes showed higher circularity and lower area, proving the principles of enrichment of a more homogenous cell population with better characteristics. We proved the NEEGA-DF method is a "gentle" cell sorter that purifies primary cells obtained by enzymatic digestion and does not alter any stem cell function

Investigating elevational gradients of species richness in a Mediterranean plant hotspot using a published flora

The Apuan Alps are one of the most peculiar mountain chain in the Mediterranean, being very close to the coastline and reaching an elevation of almost 2000 m. Based on published flora, we investigated the distribution of plant species richness along the whole elevational gradient of this chain considering: (i) all species, (ii) endemic versus alien species; and (iii) functional groups of species based on Raunkiær life forms (RLF). Generalized Linear Models (GLMs) were used to analyse richness patterns along the elevational gradient, and elevational richness models versus the area of the elevational belts were fitted to test the effect of surface area. Our results showed decreasing species richness with increasing elevation. In contrast, endemic species richness increased along the elevational gradient. Alien species were mainly distributed at low elevations, but this result should be taken with caution since we used historical data. Species life forms were not equally distributed along the elevation gradient: chamaephytes and hemicryptophytes were the richest groups at high elevations, while therophytes showed highest species richness at low elevations. Our findings suggest that in the Apuan Alps there is a major elevational gradient in species composition that could reflect plant evolutionary history. Furthermore, we highlight the key role of published floras as a relevant source of biodiversity data.publishedVersio

AMS-VegBank: a new database of vegetation plots for the Italian territory

The importance of collection, storage and exchange of georeferenced vegetation plot-based data has significantly grown in the recent decades, because of the new potentialities offered by ecoinformatics. In this article we introduce the Alma Mater Studiorum – University of Bologna vegetation database (AMS-VegBank; GIVD code EU-IT-021) compiling 17,505 georeferenced vegetation-plot observations within a time span of 90 years. This database includes 337,799 occurrence data of vascular plant species, belonging to many different habitat types. The historical relevance of the presented database is highlighted by the presence of some of the most ancient vegetation-plot observations in Europe (years 1930–1938). The geographic coverage of the database is mostly for Italian territory but it includes also data from other countries. The thematic focuses represented in the database are various, such as small Mediterranean islands, the Dolomite Mountains and the Italian National Parks. The large amount of historical plots available for the country not previously included in existing databases, combined with the constant action to improve the georeferencing of existing data and the addition of new data, highlight the uniqueness of this database. AMS-VegBank represents thus an important tool for studying plant biodiversity within the context of continental and global vegetation plot databases

A Wireless Method for Beam Coupling Impedance Measurements of the LHC Goniometer

The beam coupling impedance (BCI) of an accelerator component should be ideally evaluated exciting the device with the beam itself. However, this scenario is not always attainable and alternative methods must be exploited, such as the bench measurements techniques. The stretched Wire Method (WM) is a well established technique for BCI evaluations, although nowadays its limitations are well known. In particular, the stretched wire perturbs the electromagnetic boundary conditions. Therefore, the results obtained could be inaccurate, especially below the cut-off frequency of the beam pipe in the case of cavity-like structures. To overcome these limitations, efforts are being made to investigate alternative bench measurement techniques that will not require the modification of the device under test (DUT). In this framework, a wireless method has been identified and tested for a pillbox cavity. Its potential for more complex structures, such as the LHC crystal goniometer is explored

Bead-Pull Method for transverse impedance measurements

Beam Coupling Impedance represents a serious issue in High-Energy particle accelerators. Hence there is the need to measure the impedance of a device. However, standard methods have known limitations, therefore alternative methods are considered. In this report the bead-pull method is presented to measure the longitudinal impedance and its potential regarding the transverse impedance is also explored and discussed