“Il privilegio della parola scritta”: studi, progetti e nuovi approcci metodologici per i fondi d’autore (Università degli studi di Salerno, 10-12 aprile 2019)

The essay proposes a reflection on the topics covered during the conference “The privilege of the written word: management, preservation and improvement of personal cards and books” organized by the Department of Sciences of Cultural Heritage of the University of Salerno, with the collaboration of the Library Centre of the University of Bologna, the Department of Cultural Heritage of the University of Bologna and the National Commission of Special Libraries, Archives and Author Libraries of the Italian Libraries Association. Risks are analyzed, such as those related to the dispersion and fragmentation of materials during the life of the producer, good practices, with the presentation of concrete projects, the approach to new digital technologies. The conference days, organized in sessions, were divided by themes: “Books, papers, maps: the personal workshops of writing and the arts”; “Personal funds in excavation, description and management activities”; “Personal funds between physical space and digital space: the paths of access and enhancement”. Part of the event was dedicated to the presentation of the Guidelines on the treatment of personal funds. The publication of the proceedings will certainly encourage new reflections and a remodeling of the same guidelines, also in consideration of the documentary typologies to be treated that were subject of deepening and report during these study days.Il saggio propone una riflessione sulle tematiche emerse durante il convegno “Il privilegio della parola scritta: gestione, conservazione e valorizzazione di carte e libri di persona” organizzato dal Dipartimento di scienze del patrimonio culturale dell’Università di Salerno, con la collaborazione del Centro bibliotecario dell’ateneo salernitano, il Dipartimento di beni culturali dell’Università di Bologna e la Commissione nazionale biblioteche speciali, archivi e biblioteche d’autore dell’Associazione italiana biblioteche. Vengono analizzati i rischi, quali quelli relativi alla dispersione e frammentazione dei materiali durante la vita del soggetto produttore, le buone pratiche, con la presentazione di progetti concreti, l’approccio alle nuove tecnologie digitali. Le giornate, organizzate in sessioni sono state divise per temi: “Libri, carte, carteggi: le officine personali della scrittura e delle arti”; “I fondi di persona nelle attività di scavo, descrizione e gestione”; “I fondi di persona tra spazio fisico e spazio digitale: i percorsi dell’accesso e della valorizzazione”. Una parte dell’evento è stata dedicata alla presentazione delle Linee guida sul trattamento dei fondi personali. La pubblicazione degli atti incoraggerà sicuramente nuove riflessioni e una rimodulazione delle stesse linee guida, anche in considerazione delle tipologie documentarie da trattare che sono state oggetto di approfondimento e relazione in queste giornate di studio

Gene expression profile of epithelial cells and mesenchymal cells derived from limbal explant culture

Purpose: Limbal stem cell deficiency is a challenging clinical problem and the current treatment involves replenishing the depleted limbal stem cell (LSC) pool by either limbal tissue transplantation or use of cultivated limbal epithelial cells (LEC). Our experience of cultivating the LEC on denuded human amniotic membrane using a feeder cell free method, led to identification of mesenchymal cells of limbus (MC-L), which showed phenotypic resemblance to bone marrow derived mesenchymal stem cells (MSC-BM). To understand the transcriptional profile of these cells, microarray experiments were carried out.Methods: RNA was isolated from cultured LEC, MC-L and MSC-BM and microarray experiments were carried out by using Agilent chip (4x44 k). The microarray data was validated by using Realtime and semiquntitative reverse transcription polymerase chain reaction. Results: The microarray analysis revealed specific gene signature of LEC and MC-L, and also their complementary role related to cytokine and growth factor profile, thus supporting the nurturing roles of the MC-L. We have also observed similar and differential gene expression between MC-L and MSC-BM.Conclusions: This study represents the first extensive gene expression analysis of limbal explant culture derived epithelial and mesenchymal cells and as such reveals new insight into the biology, ontogeny, and in vivo function of these cells

Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker

Abstract Limbal melanocytes (LM) are located in the basal epithelial layer of the corneoscleral limbus and interact with adjacent limbal epithelial progenitor cells. The exploration of their biological role in the maintenance of the limbal stem cell niche has been limited by the difficulty of LM isolation and cultivation. Here, we report on a facile protocol for the efficient isolation and enrichment of pure populations of human LMs by fluorescence-activated cell sorting (FACS) using antibodies raised against the cell surface marker CD117 (c-Kit). The enriched LMs retain self-renewal capacity and sustained melanin production, and are suitable to study the potential of LMs in stem cell-based corneal tissue engineering

A Decellularized Human Limbal Scaffold for Limbal Stem Cell Niche Reconstruction

The transplantation of ex vivo expanded limbal epithelial progenitor cells (LEPCs) on amniotic membrane or fibrin gel is an established therapeutic strategy to regenerate the damaged corneal surface in patients with limbal stem cell deficiency (LSCD), but the long-term success rate is restricted. A scaffold with niche-specific structure and extracellular matrix (ECM) composition might have the advantage to improve long-term clinical outcomes, in particular for patients with severe damage or complete loss of the limbal niche tissue structure. Therefore, we evaluated the decellularized human limbus (DHL) as a biomimetic scaffold for the transplantation of LEPCs. Corneoscleral tissue was decellularized by sodium deoxycholate and deoxyribonuclease I in the presence or absence of dextran. We evaluated the efficiency of decellularization and its effects on the ultrastructure and ECM composition of the human corneal limbus. The recellularization of these scaffolds was studied by plating cultured LEPCs and limbal melanocytes (LMs) or by allowing cells to migrate from the host tissue following a lamellar transplantation ex vivo. Our decellularization protocol rapidly and effectively removed cellular and nuclear material while preserving the native ECM composition. In vitro recellularization by LEPCs and LMs demonstrated the good biocompatibility of the DHL and intrastromal invasion of LEPCs. Ex vivo transplantation of DHL revealed complete epithelialization as well as melanocytic and stromal repopulation from the host tissue. Thus, the generated DHL scaffold could be a promising biological material as a carrier for the transplantation of LEPCs to treat LSCD

Transcription factor profiling identifies Sox9 as regulator of proliferation and differentiation in corneal epithelial stem/progenitor cells

Understanding transcription factor (TF) regulation of limbal epithelial stem/progenitor cells (LEPCs) may aid in using non-ocular cells to regenerate the corneal surface. This study aimed to identify and characterize TF genes expressed specifically in LEPCs isolated from human donor eyes by laser capture microdissection. Using a profiling approach, preferential limbal expression was found for SoxE and SoxF genes, particularly for Sox9, which showed predominantly cytoplasmic localization in basal LEPCs and nuclear localization in suprabasal and corneal epithelial cells, indicating nucleocytoplasmic translocation and activation during LEPC proliferation and differentiation. Increased nuclear localization of Sox9 was also observed in activated LEPCs following clonal expansion and corneal epithelial wound healing. Knockdown of SOX9 expression in cultured LEPCs by RNAi led to reduced expression of progenitor cell markers, e.g. keratin 15, and increased expression of differentiation markers, e.g. keratin 3. Furthermore, SOX9 silencing significantly suppressed the proliferative capacity of LEPCs and reduced levels of glycogen synthase kinase 3 beta (GSK-3ß), a negative regulator of Wnt/ß-catenin signaling. Sox9 expression, in turn, was significantly suppressed by treatment of LEPCs with exogenous GSK-3ß inhibitors and enhanced by small molecule inhibitors of Wnt signaling. Our results suggest that Sox9 and Wnt/ß-catenin signaling cooperate in mutually repressive interactions to achieve a balance between quiescence, proliferation and differentiation of LEPCs in the limbal niche. Future molecular dissection of Sox9-Wnt interaction and mechanisms of nucleocytoplasmic shuttling of Sox9 may aid in improving the regenerative potential of LEPCs and the reprogramming of non-ocular cells for corneal surface regeneration

A decellularized human corneal scaffold for anterior corneal surface reconstruction

Abstract Allogenic transplants of the cornea are prone to rejection, especially in repetitive transplantation and in scarred or highly vascularized recipient sites. Patients with these ailments would particularly benefit from the possibility to use non-immunogenic decellularized tissue scaffolds for transplantation, which may be repopulated by host cells in situ or in vitro. So, the aim of this study was to develop a fast and efficient decellularization method for creating a human corneal extracellular matrix scaffold suitable for repopulation with human cells from the corneal limbus. To decellularize human donor corneas, sodium deoxycholate, deoxyribonuclease I, and dextran were assessed to remove cells and nuclei and to control tissue swelling, respectively. We evaluated the decellularization effects on the ultrastructure, optical, mechanical, and biological properties of the human cornea. Scaffold recellularization was studied using primary human limbal epithelial cells, stromal cells, and melanocytes in vitro and a lamellar transplantation approach ex vivo. Our data strongly suggest that this approach allowed the effective removal of cellular and nuclear material in a very short period of time while preserving extracellular matrix proteins, glycosaminoglycans, tissue structure, and optical transmission properties. In vitro recellularization demonstrated good biocompatibility of the decellularized human cornea and ex vivo transplantation revealed complete epithelialization and stromal repopulation from the host tissue. Thus, the generated decellularized human corneal scaffold could be a promising biological material for anterior corneal reconstruction in the treatment of corneal defects

A carotenoid-deficient mutant in Pantoea sp. YR343, a bacteria isolated from the Rhizosphere of Populus deltoides, is defective in root colonization

The complex interactions between plants and their microbiome can have a profound effect on the health and productivity of the plant host. A better understanding of the microbial mechanisms that promote plant health and stress tolerance will enable strategies for improving the productivity of economically important plants. Pantoea sp. YR343 is a motile, rod-shaped bacterium isolated from the roots of Populus deltoides that possesses the ability to solubilize phosphate and produce the phytohormone indole-3-acetic acid (IAA). Pantoea sp. YR343 readily colonizes plant roots and does not appear to be pathogenic when applied to the leaves or roots of selected plant hosts. To better understand the molecular mechanisms involved in plant association and rhizosphere survival by Pantoea sp. YR343, we constructed a mutant in which the crtB gene encoding phytoene synthase was deleted. Phytoene synthase is responsible for converting geranylgeranyl pyrophosphate to phytoene, an important precursor to the production of carotenoids. As predicted, the ΔcrtB mutant is defective in carotenoid production, and shows increased sensitivity to oxidative stress. Moreover, we find that the ΔcrtB mutant is impaired in biofilm formation and production of IAA. Finally we demonstrate that the ΔcrtB mutant shows reduced colonization of plant roots. Taken together, these data suggest that carotenoids are important for plant association and/or rhizosphere survival in Pantoea sp. YR343.Work at the University of Notre Dame was supported by DOE grant SC0006642 (RM) and by a subcontract from Oak Ridge National Laboratory (SP).http://www.frontiersin.orgam2016Microbiology and Plant Patholog

Hidden Markov Chain Analysis: Impact of Misclassification on Effect of Covariates in Disease Progression and Regression

Most of the chronic diseases have a well-known natural staging system through which the disease progression is interpreted. It is well established that the transition rates from one stage of disease to other stage can be modeled by multi state Markov models. But, it is also well known that the screening systems used to diagnose disease states may subject to error some times. In this study, a simulation study is conducted to illustrate the importance of addressing for misclassification in multi-state Markov models by evaluating and comparing the estimates for the disease progression Markov model with misclassification opposed to disease progression Markov model. Results of simulation study support that models not accounting for possible misclassification leads to bias. In order to illustrate method of accounting for misclassification is illustrated using dementia data which was staged as no cognitive impairment, mild cognitive impairment and dementia and diagnosis of dementia stage is prone to error sometimes. Subjects entered the study irrespective of their state of disease and were followed for one year and their disease state at follow up visit was recorded. This data is used to illustrate that application of multi state Markov model which is an example of Hidden Markov model in accounting for misclassification which is based on an assumption that the observed (misclassified) states conditionally depend on the underlying true disease states which follow the Markov process. The misclassification probabilities for all the allowed disease transitions were also estimated. The impact of misclassification on the effect of covariates is estimated by comparing the hazard ratios estimated by fitting data with progression multi state model and by fitting data with multi state model with misclassification which revealed that if misclassification has not been addressed the results are biased. Results suggest that the gene apoe ε4 is significantly associated with disease progression from mild cognitive impairment to dementia but, this effect was masked when general multi state Markov model was used. While there is no significant relation is found for other transitions

About the Contemporary Public Library. A Short Selective Bibliography

This short work was drawn up after the Turin conference (12 December, 2015), in which speakers discussed very important issues, all related to the theme of the identity of the contemporary public library. With this paper, my purpose is limited and modest, and offers a short survey of publications in Italy, Spain and France concerning the topics discussed at the conference, and presents a brief selective bibliography. I believe that this tool will be useful to broaden the debate. I would like ..

Physical Layer Security With Active Jamming Using NOMA.

This paper is persuaded to understand the physical layer security in wireless commu-nications utilizing NOMA (Non Orthogonal Multiple Access) concepts in the presence of an eavesdropper. Physical layer security maintains the confidentiality and secrecyof the system against eavesdroppers. We use the power domain in this paper, where NOMA allows many users to share resources side by side. Power allocation concern-ing channel condition is taken into consideration where user whose channel condition is weak is allocated with eminent power to directly decode the signal, whereas theuser with better channel condition applies successive interference cancellation (SIC)to decode the signal. Here, the base station communicates with the users and sends data signals while the eavesdropper secretly eavesdrops on the confidential informa-tion simultaneously. In this thesis, to improve the physical layer security, jamming method was usedwhere users are assumed to be in full duplex, send jamming signals to degrade the performance of the eavesdropper. Analytic expressions of CDF, PDF, outage proba-bility and secrecy capacity are obtained from analyzing the NOMA jamming scheme. The numerical results are evaluated with the simulations results and analysed theeffect of jamming on improving the performance of the NOMA system in presenceof an eavesdropper