Three-dimension-printed custom-made prosthetic reconstructions: from revision surgery to oncologic reconstructions

Background The use of custom-made 3D-printed prostheses for reconstruction of severe bone defects in selected cases is increasing. The aims of this study were to evaluate (1) the feasibility of surgical reconstruction with these prostheses in oncologic and non-oncologic settings and (2) the functional results, complications, and outcomes at short-term follow-up. Methods We analyzed 13 prospectively collected patients treated between June 2016 and January 2018. Diagnoses were primary bone tumour (7 patients), metastasis (3 patients), and revision of total hip arthroplasty (3 patients). Pelvis was the most frequent site of reconstruction (7 cases). Functional results were assessed with MSTS score and complications according to Henderson et al. Statistical analysis was performed using Kaplan-Meier and log-rank test curves. Results At a mean follow-up of 13.7 months (range, 6 \u2013 26 months), all patients except one were alive. Oncologic outcomes show seven patients NED (no evidence of disease), one NED after treatment of metastasis, one patient died of disease, and another one was alive with disease. Overall survival was 100% and 80% at one and two years, respectively. Seven complications occurred in five patients (38.5%). Survival to all complications was 62% at two years of follow-up. Functional outcome was good or excellent in all cases with a mean score of 80.3%. Conclusion 3D-printed custom-made prostheses represent a promising reconstructive technique in musculoskeletal oncology and challenging revision surgery. Preliminary results were satisfactory. Further studies are needed to evaluate prosthetic design, fixation methods, and stability of the implants at long-ter

EMT/MET at the crossroad of stemness, regeneration and oncogenesis. The Ying-Yang equilibrium recapitulated in cell spheroids

The epithelial-to-mesenchymal transition (EMT) is an essential trans-differentiation process, which plays a critical role in embryonic development, wound healing, tissue regeneration, organ fibrosis, and cancer progression. It is the fundamental mechanism by which epithelial cells lose many of their characteristics while acquiring features typical of mesenchymal cells, such as migratory capacity and invasiveness. Depending on the contest, EMT is complemented and balanced by the reverse process, the mesenchymal-to-epithelial transition (MET). In the saving economy of the living organisms, the same (Ying-Yang) tool is integrated as a physiological strategy in embryonic development, as well as in the course of reparative or disease processes, prominently fibrosis, tumor invasion and metastasis. These mechanisms and their related signaling (e.g., TGF-β and BMPs) have been effectively studied in vitro by tissue-derived cell spheroids models. These three-dimensional (3D) cell culture systems, whose phenotype has been shown to be strongly dependent on TGF-β-regulated EMT/MET processes, present the advantage of recapitulating in vitro the hypoxic in vivo micro-environment of tissue stem cell niches and their formation. These spheroids, therefore, nicely reproduce the finely regulated Ying-Yang equilibrium, which, together with other mechanisms, can be determinant in cell fate decisions in many pathophysiological scenarios, such as differentiation, fibrosis, regeneration, and oncogenesis. In this review, current progress in the knowledge of signaling pathways affecting EMT/MET and stemness regulation will be outlined by comparing data obtained from cellular spheroids systems, as ex vivo niches of stem cells derived from normal and tumoral tissues. The mechanistic correspondence in vivo and the possible pharmacological perspective will be also explored, focusing especially on the TGF-β-related networks, as well as others, such as SNAI1, PTEN, and EGR1. This latter, in particular, for its ability to convey multiple types of stimuli into relevant changes of the cell transcriptional program, can be regarded as a heterogeneous "stress-sensor" for EMT-related inducers (growth factor, hypoxia, mechano-stress), and thus as a therapeutic target

Analysis of pregnancy-associated plasma protein a production in human adult cardiac progenitor cells

IGF-binding proteins (IGFBPs) and their proteases regulate IGFs bioavailability in multiple tissues. Pregnancy-associated plasma protein A (PAPP-A) is a protease acting by cleaving IGFBP2, 4, and 5, regulating local bioavailability of IGFs. We have previously shown that IGFs and IGFBPs are produced by human adult cardiac progenitor cells (haCPCs) and that IGF-1 exerts paracrine therapeutic effects in cardiac cell therapy with CPCs. Using immunofluorescence and enzyme immunoassays, we firstly report that PAPP-A is produced and secreted in surprisingly high amounts by haCPCs. In particular, the homodimeric, enzymatically active, PAPP-A is secreted in relevant concentrations in haCPC-conditioned media, while the enzymatically inactive PAPPA/proMBP complex is not detectable in the same media. Furthermore, we show that both homodimeric PAPP-A and proMBP can be detected as cell associated, suggesting that the previously described complex formation at the cell surface does not occur easily, thus positively affecting IGF signalling. Therefore, our results strongly support the importance of PAPP-A for the IGFs/IGFBPs/PAPP-A axis in CPCs biology

A Smart Watch with Embedded Sensors to Recognize Objects, Grasps and Forearm Gestures

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Pseudomonas protegens MP12: A plant growth-promoting endophytic bacterium with broad-spectrum antifungal activity against grapevine phytopathogens

Abstract Pseudomonas sp. MP12 was isolated from a soil sample collected in a typical warm-temperate deciduous forest near Brescia, Northern Italy. Phylogenetic analysis identified the species as Pseudomonas protegens. We evidenced in this strain the presence of the genes phlD, pltB and prnC responsible for the synthesis of the antifungal compounds 2,4-diacetylphloroglucinol (2,4-DAPG), pyoluteorin and pyrrolnitrin, respectively. P. protegens MP12 was also shown to produce siderophores and ammonia, yielded positive results with the indole-3-acetic acid test, and was capable of phosphate solubilization. Moreover, P. protegens MP12 exhibited inhibitory effects on in vitro mycelial growth of prominent grapevine (Vitis vinifera) phytopathogens such as Botrytis cinerea, Alternaria alternata, Aspergillus niger, Penicillium expansum and Neofusicoccum parvum. The strain showed activity even against Phaeomoniella chlamydospora and Phaeoacremonium aleophilum, which cause the devastating tracheomycosis/esca disease of grapevine trunks for which no efficacious control methods have been demonstrated so far. Furthermore, the MP12 strain manifested in vivo antifungal activity against B. cinerea on grapevine leaves. Culture-dependent and culture-independent analysis revealed the ability of P. protegens MP12 to efficiently and permanently colonize inner grapevine tissues. These results suggest that P. protegens MP12 could be worth of exploitation as an antifungal biocontrol agent for applications in viticulture

In vivo endophytic, rhizospheric and epiphytic colonization of vitis vinifera by the plant-growth promoting and antifungal strain pseudomonas protegens MP12

An evaluation was conducted of the colonization of Pseudomonas protegens MP12, a plantgrowth promoting and antagonistic strain, inoculated in vine plants during a standard process of grapevine nursery propagation. Three in vivo inoculation protocols (endophytic, rhizospheric, and epiphytic) were implemented and monitored by means of both culture-dependent and independent techniques. Endophytic treatment resulted in the colonization of the bacterium inside the vine cuttings, which spread to young leaves during the forcing period. Microscopy analysis performed on transformed dsRed-tagged P. protegens MP12 cells confirmed the bacterium’s ability to penetrate the inner part of the roots. However, endophytic MP12 strain was no longer detected once the plant materials had been placed in the vine nursery field. The bacterium also displayed an ability to colonize the rhizosphere and, when the plants were uprooted at the end of the vegetative season, its persistence was confirmed. Epiphytic inoculation, performed by foliar spraying of cell suspension, was effective in controlling artificially-induced Botrytis cinerea infection in detached leaves. The success of rhizospheric and leaf colonization in vine plants suggests potential for the future exploitation of P. protegens MP12 as biofertilizer and biopesticide. Further investigation is required into the stability of the bacterium’s colonization of vine plants under real-world conditions in vineyards

The Effects of the COVID-19-induced Lockdown on the Social Capital and Cultural Capital in Italy

The present study investigated the effects of the first COVID-19 lockdown on the Cultural and Social Capitals in Italy in a large group of adults (n = 1125). The relationships between the COVID-19 spread and participants’ Cultural Capital, Social Capital, educational level, occupational prestige, and age were studied using structural equation models. For women but not for men, pandemic spread was positively affected by occupational prestige and it had a positive relationship with their Social Capital (women: CFI = 0.949; RMSEA = 0.059 [CI = 0.045-0.075]; men: CFI = 0.959; RMSEA = 0.064 [CI = 0.039–0.087]). Moreover, the participants were divided into three validated clusters based on their Cultural and Social Capitals levels to investigate changes in the Capitals compared with the pre-lockdown period. It was found that the lockdown contributed to improving the gap among individuals increasing high levels and decreasing low levels of both the Capitals. People with high Cultural and Social Capitals seemed to have seized the opportunity given by COVID-19 restrictions to cultivate their cultural interests and become more involved within their networks. In contrast, individuals with low Cultural and Social Capitals paid the highest price for the social isolation. Given that the Capitals encourage healthy behavior and influence well-being and mental health, institutions should develop or improve their policies and practices to foster individual resources, and make fairer opportunities available during the pandemic

Normal versus pathological cardiac fibroblast-derived extracellular matrix differentially modulates cardiosphere-derived cell paracrine properties and commitment

Human resident cardiac progenitor cells (CPCs) isolated as cardiosphere-derived cells (CDCs) are under clinical evaluation as a therapeutic product for cardiac regenerative medicine. Unfortunately, limited engraftment and differentiation potential of transplanted cells significantly hamper therapeutic success. Moreover, maladaptive remodelling of the extracellular matrix (ECM) during heart failure progression provides impaired biological and mechanical signals to cardiac cells, including CPCs. In this study, we aimed at investigating the differential effect on the phenotype of human CDCs of cardiac fibroblast-derived ECM substrates from healthy or diseased hearts, named, respectively, normal or pathological cardiogel (CG-N/P). After 7 days of culture, results show increased levels of cardiogenic gene expression (NKX2.5, CX43) on both decellularized cardiogels compared to control, while the proportion and staining patterns of GATA4, OCT4, NKX2.5, ACTA1, VIM, and CD90-positive CPCs were not affected, as assessed by immunofluorescence microscopy and flow cytometry analyses. Nonetheless, CDCs cultured on CG-N secreted significantly higher levels of osteopontin, FGF6, FGF7, NT-3, IGFBP4, and TIMP-2 compared to those cultured on CG-P, suggesting overall a reduced trophic and antiremodelling paracrine profile of CDCs when in contact with ECM from pathological cardiac fibroblasts. These results provide novel insights into the bidirectional interplay between cardiac ECM and CPCs, potentially affecting CPC biology and regenerative potential

THE STUDY OF THREE GRAPEVINE CLONES TO UNCOVER THE GENETIC TRAITS RESPONSIBLE FOR THE LOW SUSCEPTIBILITY TO FLAVESCENCE DORéE

Flavescence dorée (FD) is one of the most destructive grapevine yellows diseases and a quarantine pest in the European Community. It is caused by phytoplasmas, which are transmitted in vineyard by the leafhopper Scaphoideus titanus. Inter and intraspecific differences in susceptibility to FD have already been observed among grapevine varieties and clones of the same variety. Grapevine varieties and clones completely resistant to FD have not been uncovered yet, however these differences suggest the presence of genetic traits in grapevine related to high or low susceptibility to FD. Cultivated grapevines are clonally propagated and the genome of each cultivar is preserved, except for the accumulation of mutations over time that can generate distinguishable clones with several notable phenotypes. The differences between the genome of clones of the same variety are less than those between different varieties; thus, in order to highlight the genetic features responsible for the different phenotypes, the study of the genomes of clones with different susceptibility to FD can be a valid technique to achieve the goal. The aim of this work is to find out the genetic traits responsible for the different susceptibility to FD among three Chardonnay clones, analyzing the diversity in their genomes and transcriptomic profiles. The clones were sequenced by accurate whole genome techniques, Hi-Fi reads sequencing on PacBio platform coupled with Illumina, and then the genomes were de novo assembled. Moreover, two of them were experimentally infected in field by means of the insect vector and the transcriptomic profiles in the early stage of FD infection were analyzed. In particular, the clones were compared in absence of the disease and the vector, in presence of the healthy vector and with the FD infective vector. The results obtained from the preliminary genome comparison showed higher similarity between the three clones than compared with the reference genome from variety Pinot noir, while the analysis to identify the genomic differences among clones are still in progress. The transcriptomic profiles showed interesting differences in some pathways expressed in presence of healthy vectors, while the clones shared a similar expression profile before being in contact with the vectors as well as after the challenge with the FD infective vectors. These findings might suggest the involvement of antibiosis mechanisms in the partial resistance of the specific Chardonnay clone to FD. The results obtained, and those that will be achieved in future, are useful for new breeding programs and clonal selection. Grapevine plants more resistant to FD will permit to decrease the insecticides used to control the disease in vineyards. Moreover, the knowledge of the molecular and metabolic mechanisms involved in scarcely susceptible clones could pave the way for the development, for example, of biostimulants capable to induce such defenses even in the most susceptible plants