The unbearable lightness of bone marrow homeostasis

The anatomical and functional dimensions of bone marrow topography have been at the forefront of modern bone and immunological research for many years and remain a source of complexity and perplexity due to the multitude of microhabitats within this microenvironment. In fact, research has uncovered fascinating functional aspects of bone marrow residents, and the bone marrow niche has been identified as the foremost reservoir of a variety of cells including hematopoietic, skeletal and endothelial stem/progenitor cells. The physical interactions of the marrow residents, combined with the release of cytokines and growth factors, organize well-defined operative compartments, which preserve bone and immune cell homeostasis. In a simplistic view, both the hematopoietic and bone marrow stromal (mesenchymal) stem/progenitor cell populations dwell at the interface between the endosteum and the bone marrow area (endosteal niche) and in the perivascular space (vascular niche). Indeed, the tantalizing hypothesis of bone marrow regulatory dependency on these niches is supported by current research insofar as the increase in the number of osteoblasts results in a concomitant increase in the hematopoietic population, indicating that the osteoblasts and the endosteal niche are key components of HSC maintenance. On the other hand, impaired function of the vascular niche compromises the endosteal niche's ability to support hematopoiesis. These fascinating discoveries indicate that there are strong ties between bone marrow inhabitants within the confines of the bone marrow itself. When these ties fail, niche-niche communication suffers and results in reduced bone formation, enfeebled hematopoiesis and unrestrained HSC migration through blood circulation. This study focused on the extraordinary homeostatic equilibrium and function of both bone and immune cells within the spatially defined microenvironment of bone marrow. But how important is the anatomically outlined scenery in which the bone marrow entity supports and hosts the hematopoietic elements

Aspetti evolutivi del Knowledge Management: considerazioni teoriche ed applicazioni operative. L'analisi del caso Loccioni

Negli ultimi anni si è assistito ad un impercettibile e progressivo cambiamento: l’economia dei nostri giorni sta diventando un’economia cognitiva, entrando a far parte della knowledge era. Si è verificato, cioè, un ribaltamento che ha modificato il tradizionale modo di intendere i fattori critici di successo: è emerso come nuovo elemento la gestione integrata di tutti quei saperi utili per il coordinamento delle attività da parte dei soggetti che operano all'interno delle organizzazioni. Occorre dotarsi, quindi, di strumenti che sappiano amministrare la risorsa conoscenza, trasformando le organizzazioni in organizzazioni che apprendono. Si tratta, in definitiva, di predisporre un idoneo sistema di Knowledge Management (KM) che possa conferire alle imprese tutti i vantaggi che derivano da pratiche di condivisione della conoscenza. Il presente lavoro si inserisce nel filone di ricerca del Knowledge Management con l’obiettivo di fornire una chiave di lettura critica ai fenomeni di creazione, diffusione e sfruttamento delle conoscenze nell'ambito delle dinamiche di sviluppo di grande impresa industriale e di comprendere sotto quali condizioni organizzative e mediante quale tipo di approccio l’investimento in un sistema di Knowledge Management sia produttivo di effetti positivi. Tale lavoro di tesi, è stato sviluppato a partire dall'analisi empirica, che adotta un approccio qualitativo con finalità esplorative, al fine di suggerire delle prime ipotesi di comprensione dei fenomeni (Fattore, 2005). Sulla base di ciò è stata adottata la metodologia dello studio di caso, quale quello del Gruppo Loccioni, al fine di comprendere e dimostrare come l’implementazione di un sistema di KM non rappresenti un processo lineare e che risulta necessario esplorare il contesto di riferimento e, in particolare, le caratteristiche organizzative dell’impresa, per poter valutare quale modello sia più adatto ad essere applicato per non rischiare il fallimento del progetto.In recent years we have witnessed an imperceptible and progressive change: nowadays, economy is becoming cognitive, being part of the knowledge era. So, a reversal occurred which has changed the traditional understanding of the critical success’s factors: the integrated management has emerged, as a new element of all the knowledge useful for the coordination of activities by parties which operate within organizations. It is necessary to have, therefore, tools that are able to manage the "knowledge" resource, transforming organizations into learning organizations. This means, ultimately, to set up an appropriate system of Knowledge Management (KM) which can give to companies all the advantages as a result of knowledge sharing practices. This work is part of the research field of Knowledge Management with the aim of providing a critical key to make sure to understand the creation, dissemination and exploitation of knowledge phenomena in the dynamics of development of a large industrial company and to understand under what organizational conditions and by what type of approach, investing in a Knowledge Management system, produce its positive effects. This doctoral thesis has been developed starting from the empirical analysis, following a qualitative approach for exploratory purposes, with the purpose of suggesting some of the first hypothesis of understanding (Fattore, 2005). The methodology of the case study, such as the Loccioni Group, was adopted in order to understand and demonstrate how the implementation of a KM system does not represent a linear process and which is necessary to explore the surrounding context, in particular the organizational characteristics of the company, in order to evaluate which model is most suitable to be applied for not to risk the failure of the project

I nuovi confini della responsabilità patrimoniale del debitore

This doctoral thesis is dedicated to the topic of the debtor’s responsibility, considered from the point of view of its evolution in the contemporary legal experience. Traditionally, the system of debtor’s responsibility had been represented as it was based on two dogmas that seemed to be insuperable. The first one identify itself in uniqueness and indivisibility of the estate and in the universal patrimonial liability principle ex article 2740 c.c. of the Italian Civil Code, destined to give out only in peremptory cases established by the law. The second one is represented by the nullity of conventional limitations of debtor’s patrimonial liability. Within this framework, it was not only often affirmed that all debtor’s assets are included into the generic patrimonial guarantee and that its limitations are exceptions, having residual character that only the law can establish, but it was also precluded to the private autonomy to interfere with this institution. The article 2740 c.c. seems to confirm this reconstruction, that, however, must be confronted with the normative evolution, that is from several years directed towards the opposite direction. This evolution is especially due to the proliferation of the cases in which the law allows to the same subject to have more than one estate, that had put into crisis the idea according to which the debtor must guarantee his fulfilment with all his assets. The objective of this thesis is to rebuild the new boundaries of debtor’s patrimonial liability, taking into account legal innovations, as well as doctrinal and jurisprudential evolution. The first question to be answered by this study regards to the effectiveness of the dogmas that are traditionally considered the foundations of the whole system, trying to verify if they were really imposed by insuperable principles or if our system has always had the basis to suggest a different key of interpretation. This work is dedicated in its first part to exploring the concept of patrimonial liability in its historical developments up to the present codification. Within this thesis’ part would be explained the reasons why this institution is considered one of the most important pillars of our legal system and it would be put the basis to a different reading key of the system, occasioned also by the comparison with the approach to the theme of segregated found developed in Scotland and Québec. In the second part of this work it would be explored some examples of segregated founds that are present in our legal system, as fondo patrimoniale (articles 167 c.c.); patrimoni destinati a uno specifico affare (articles 2447 bis c.c.) and atto di destinazione ex article 2645 ter c.c. In this way it would be possible to verify how they had contribute to the evolution of the system and it would be possible to determine the new boundaries of debtor’s patrimonial liability. The last part of this thesis is dedicated to exploring the dogma that determined the nullity of conventional limitations of debtor’s patrimonial liability, whose legal basis is represented by Article 2740, paragraph 2, of the Italian Civil Code. The fundamental problem is to understand if this dogma had been really imposed by this rule or if it was dictated by doctrinal elaboration. From this point of view, the analysis wants to verify if the reasons that make it necessary the typicality principle of segregated found remain valid also for conventional limitations of debtor’s patrimonial liability

Degradation rate of 5-fluorouracil in metastatic colorectal cancer. A new predictive outcome biomarker?

BACKGROUND: 5-FU based chemotherapy is the most common first line regimen used for metastatic colorectal cancer (mCRC). Identification of predictive markers of response to chemotherapy is a challenging approach for drug selection. The present study analyzes the predictive role of 5-FU degradation rate (5-FUDR) and genetic polymorphisms (MTHFR, TSER, DPYD) on survival. MATERIALS AND METHODS: Genetic polymorphisms of MTHFR, TSER and DPYD, and the 5-FUDR of homogenous patients with mCRC were retrospectively studied. Genetic markers and the 5-FUDR were correlated with clinical outcome. RESULTS: 133 patients affected by mCRC, treated with fluoropyrimidine-based chemotherapy from 2009 to 2014, were evaluated. Patients were classified into three metabolic classes, according to normal distribution of 5-FUDR in more than 1000 patients, as previously published: poor-metabolizer (PM) with 5-FU-DR ≤ 0,85 ng/ml/106 cells/min (8 pts); normal metabolizer with 0,85 < 5-FU-DR < 2,2 ng/ml/106 cells/min (119 pts); ultra-rapid metabolizer (UM) with 5-FU-DR ≥ 2,2 ng/ml/106 cells/min (6 pts). PM and UM groups showed a longer PFS respect to normal metabolizer group (14.5 and 11 months respectively vs 8 months; p = 0.029). A higher G3-4 toxicity rate was observed in PM and UM, respect to normal metabolizer (50% in both PM and UM vs 18%; p = 0.019). No significant associations between genes polymorphisms and outcomes or toxicities were observed. CONCLUSION: 5-FUDR seems to be significantly involved in predicting survival of patients who underwent 5-FU based CHT for mCRC. Although our findings require confirmation in large prospective studies, they reinforce the concept that individual genetic variation may allow personalized selection of chemotherapy to optimize clinical outcomes

Three-Dimensional Radiographic Evaluation of the Malar Bone Engagement Available for Ideal Zygomatic Implant Placement

Zygomatic implant rehabilitation is a challenging procedure that requires an accurate prosthetic and implant plan. The aim of this study was to evaluate the malar bone available for three-dimensional zygomatic implant placement on the possible trajectories exhibiting optimal occlusal emergence. After a preliminary analysis on 30 computed tomography (CT) scans of dentate patients to identify the ideal implant emergencies, we used 80 CT scans of edentulous patients to create two sagittal planes representing the possible trajectories of the anterior and posterior zygomatic implants. These planes were rotated clockwise on the ideal emergence points and three different hypothetical implant trajectories per zygoma were drawn for each slice. Then, the engageable malar bone and intra- and extra-sinus paths were measured. It was possible to identify the ideal implant emergences via anatomical landmarks with a high predictability. Significant differences were evident between males and females, between implants featuring anterior and those featuring posterior emergences, and between the different trajectories. The use of internal trajectories provided better bone engagement but required a higher intra-sinus path. A significant association was found between higher intra-sinus paths and lower crestal bone heights

Tuning the cytotoxicity of ruthenium(ii) para-cymene complexes by mono-substitution at a triphenylphosphine/phenoxydiphenylphosphine ligand

The new complexes [RuCl2(η6-p-cymene)(κP-Ph2PR)] [R = 4-C6H4OSiMe2tBu, 1; R = 4-C6H4Br, 2; R = OC(O)CHCl2, 3; R = OPh, 4; R = O(2-C6H4SiMe2tBu), 5] and [Ru(C2O4)(η6-p-cymene)κP-Ph2PO(2-C6H4(SiMe2tBu))], 6, were obtained in 83-98% yield from Ru(ii) arene precursors by three different synthetic strategies. The unprecedented phosphine Ph2P(O(2-C6H4SiMe2tBu)) was synthesized in 86% yield from 2-C6H4Br(OSiMe2tBu) and Ph2PCl, via intramolecular oxygen to carbon 1,3 migration of the silyl group (retro-Brook rearrangement). All the complexes were fully characterized by analytical and spectroscopic methods, and by single crystal X-ray diffraction in the cases of 3, 4, 5 and 6. Complexes 1-6 and the model compounds [RuCl2(η6-p-cymene)(κP-PPh3)] (Ru-PPh3) and [Ru(C2O4)(η6-p-cymene)(κP-PPh3)] (Ru-PPh3-O) underwent slow degradation in chloroform solutions upon air contact; the mixed valence complex [(η6-p-cymene)Ru(μ-Cl)3RuCl2(κP-PPh3)], 7, was isolated from a solution of Ru-PPh3in CHCl3, and X-ray identified. The antiproliferative activity of 1-6 and Ru-PPh3, Ru-PPh3-O and [RuCl2(η6-p-cymene)(κP-PTA)] (RAPTA-C) was assessed towards the triple-negative breast cancer cell line MDA-MB-231, the ovarian carcinoma cell line A2780 and human skin fibroblasts (HSF). Complexes 1, 2, 5 and 6 displayed IC50values significantly lower than that of cisplatin, with 2 providing a more potent cytotoxic effect on MDA-MB-231 and A2780 cancer cells compared to the noncancerous cell line (HSF). The stability of all complexes in DMSO/water solution was elucidated by NMR and conductivity measurements, and in particular35Cl NMR spectroscopy was helpful to check the possible chloride dissociation. The stability studies suggest that the cytotoxic activity in vitro of the compounds is mainly ascribable to Ru(ii) species still bound to the phosphorus ligand

Administration of DNA Plasmid Coding Protein Aggregating Domain Induces Inflammatory Bone Loss.

Background: Plasmids coding protein aggregation polypeptides from different sources have beenproposed as genetic adjuvants for DNA vaccines. We reported that a plasmid (pATRex), encompassing the DNA sequence for the von Willebrand A (vWA/A) domain of the Anthrax Toxin Receptor-1 (ANTXR-1, alias TEM8, Tumor Endothelial Marker 8), acts as strong immune adjuvant by inducing formation of insoluble intracellular aggregates and subsequent cell death. Aims: In the present study we addressed the question of whether there is any substantial immunotoxicity associated with the use of self-aggregating proteins as genetic adjuvants. Results: Here we report, by mean of histology, X-ray and molecular examinations of bone specimens, the unexpected finding that intramuscular injection of pATRex in mice triggers, per se, severe bone loss (osteoporosis) independently from the sex and genotype of the treated animals. Conclusion: Even though the study suggests that proteinaceous “sticky “ adjuvants are unlikely to find their way into practical vaccination, the information gained is of value as ATRex injections could provide an additional, simplified, mouse model of osteoporosis. Moreover, our results provide an experimental support to the hypothesis that proteotoxic aggregates chronically activate the innate immune system in amyloid and aggregosome disorders

Heptad stereotypy, S/Q layering, and remote origin of the SARS-CoV-2 fusion core

The fusion of the SARS-CoV-2 virus with cells, a key event in the pathogenesis of Covid-19, depends on the assembly of a six-helix fusion core (FC) formed by portions of the spike protein heptad repeats (HRs) 1 and 2. Despite the critical role in regulating infectivity, its distinctive features, origin, and evolution are scarcely understood. Thus, we undertook a structure-guided positional and compositional analysis of the SARS-CoV-2 FC, in comparison with FCs of related viruses, tracing its origin and ongoing evolution. We found that clustered amino acid substitutions within HR1, distinguishing SARS-CoV-2 from SARS-CoV-1, enhance local heptad stereotypy and increase sharply the FC serine-to-glutamine (S/Q) ratio, determining a neat alternate layering of S-rich and Q-rich subdomains along the post-fusion structure. Strikingly, SARS-CoV-2 ranks among viruses with the highest FC S/Q ratio, together with highly syncytiogenic respiratory pathogens (RSV, NDV), whereas MERS-Cov, HIV, and Ebola viruses display low ratios, and this feature reflects onto S/Q segregation and H-bonding patterns. Our evolutionary analyses revealed that the SARS-CoV-2 FC occurs in other SARS-CoV-1-like Sarbecoviruses identified since 2005 in Hong Kong and adjacent regions, tracing its origin to >50 years ago with a recombination-driven spread. Finally, current mutational trends show that the FC is varying especially in the FC1 evolutionary hotspot. These findings establish a novel analytical framework illuminating the sequence/structure evolution of the SARS-CoV-2 FC, tracing its long history within Sarbecoviruses, and may help rationalize the evolution of the fusion machinery in emerging pathogens and the design of novel therapeutic fusion inhibitors

INF-γ encoding plasmid administration triggers bone loss and disrupts bone marrow microenvironment

IFN-γ is a pleotropic cytokine produced in the bone microenvironment. Although IFN-γ is known to play a critical role on bone remodeling, its function is not fully elucidated. Consistently, outcomes on the effects of IFN-γ recombinant protein on bone loss are contradictory among reports. In our work we explored, for the first time, the role of IFN-γ encoding plasmid (pIFN-γ) in a mouse model of osteopenia induced by ovariectomy and in the sham-operated counterpart to estimate its effects in skeletal homeostasis. Ovariectomy produced a dramatic decrease of bone mineral density (BMD). pINF-γ injected mice showed a pathologic bone and bone marrow phenotype; the disrupted cortical and trabecular bone microarchitecture was accompanied by an increased release of pro-inflammatory cytokine by bone marrow cells. Moreover, mesenchymal stem cells’ (MSCs) commitment to osteoblast was found impaired, as evidenced by the decline of osterix-positive (Osx+) cells within the mid-diaphyseal area of femurs. For instance, a reduction and redistribution of CXCL12 cells have been found, in accordance with bone marrow morphological alterations. As similar effects were observed both in sham-operated and in ovariectomized mice, our studies proved that an increased IFN-γ synthesis in bone marrow might be sufficient to induce inflammatory and catabolic responses even in the absence of pathologic predisposing substrates. In addition, the obtained data might raise questions about pIFN-γ’s safety when it is used as vaccine adjuvant