Extracellular Vesicles from Plants: Current Knowledge and Open Questions

The scientific interest in the beneficial properties of natural substances has been recognized for decades, as well as the growing attention in extracellular vesicles (EVs) released by different organisms, in particular from animal cells. However, there is increasing interest in the isolation and biological and functional characterization of these lipoproteic structures in the plant kingdom. Similar to animal vesicles, these plant-derived extracellular vesicles (PDEVs) exhibit a complex content of small RNAs, proteins, lipids, and other metabolites. This sophisticated composition enables PDEVs to be therapeutically attractive. In this review, we report and discuss current knowledge on PDEVs in terms of isolation, characterization of their content, biological properties, and potential use as drug delivery systems. In conclusion, we outline controversial issues on which the scientific community shall focus the attention shortly

The dark side of foetal bovine serum in extracellular vesicle studies

Extracellular vesicles (EVs) have been shown to be involved in cell‐cell communication and to take part in both physiological and pathological processes. Thanks to their exclusive cargo, which includes proteins, lipids, and nucleic acids from the originating cells, they are gaining interest as potential biomarkers of disease. In recent years, their appealing features have been fascinating researchers from all over the world, thus increasing the number of in vitro studies focused on EV release, content, and biological activities. Cultured cell lines are the most‐used source of EVs; however, the EVs released in cell cultures are influenced by the cell culture conditions, such as the use of foetal bovine serum (FBS). FBS is the most common supplement for cell culture media, but it is also a source of contaminants, such as exogenous bovine EVs, RNA, and protein aggregates, that can contaminate the cell‐derived EVs and influence their cargo composition. The presence of FBS contaminants in cell‐derived EV samples is a well‐known issue that limits the clinical applications of EVs, thus increasing the need for standardization. In this review, we will discuss the pros and cons of using FBS in cell cultures as a source of EVs, as well as the protocols used to remove contaminants from FBS

Word Collocation Dictionary or Word Combination Dictionary? A User-oriented Perspective

Il presente contributo prende in esame alcune delle scelte metodologiche che il lessicografo che intenda a realizzare un dizionario di collocazioni si trova ad affrontare. Il lemmario deve coincidere con un determinato strato del lessico (ad es. il "vocabolario di base") oppure essere il risultato di un procedimento bottom-up che ne faccia la "proiezione" delle collocazioni effettivamente raccolte (ad es. mediante estrazione da corpora)? Considerato che le collocazioni, secondo il grado più o meno elevato di coesione e di rigidità sintattica esibite, si distribuiscono lungo un continuum delimitato da un lato dalle combinazioni libere e dall'altro dalle espressioni idiomatiche, come trattare i casi-limite? Ad esempio, che "collocazione" trovare nel dizionario per le unità polirematiche? Più in generale, quanto teorico e quanto orientato all’utente deve essere un dizionario di collocazioni? Infine, nei lemmi nominali polisemici, l’ordinamento semantico della voce deve ricalcare quello dei dizionari generali o occorre in determinati casi applicare una sorta di "lente di ingrandimento" che promuova al rango di accezioni quei sensi cui corrispondano collocati verbali specifici?O presente artigo analisa algumas das escolhas metodológicas a serem afrontadas por um lexicógrafo que pretenda elaborar um dicionário de colocações. O conjunto de entradas (nomenclatura) deve coincidir com um determinado nível do léxico (por exemplo, o “vocabulário de base”)? Ou deve ser o resultado de um procedimento bottom-up que faça a “projeção” de colocações efetivamente coletadas (por exemplo, mediante uso de corpora)? Considerando que as colocações, segundo seu grau de coesão e de rigidez sintática mais ou menos elevado, distribuem-se ao longo de um continuum delimitado, de um lado, pelas combinações livres e, de outro, pelas expressões idiomáticas, como tratar os casos-limite? Por exemplo, que colocação encontrar no dicionário para as unità polirematiche? Em termos gerais, em que medida deve ser “teórico” ou “orientado ao usuário” um dicionário de colocações? Enfim, nas entradas nominais polissêmicas, a ordem semântica do verbete deve seguir aquela dos dicionários gerais ou, em determinados casos, é necessário aplicar uma espécie de “lente de aumento” que promova à posição de acepção aqueles sentidos que selecionam colocados verbais específicos?This paper examines some of the methodological choices faced by the lexicographer wishing to compile a dictionary of collocations. Should entries exclusively come from a ‘basic vocabulary’ or should their list be the result of a ‘bottom-up’ process, i.e. an ex-post extraction of the bases of all collocations collected (e.g. by means of extraction from corpora)? In terms of cohesion and syntactic rigidity, collocations distribute themselves along a continuum whose boundaries are delimited by free combinations on the one side and by idioms on the other. Which ‘borderline’ word combinations, such as multi-word units, should be included in the dictionary? In more general terms, how theoretical and how user-oriented should a collocations dictionary be? Finally, should sense ordering in polysemous noun entries reflect that of general dictionaries or should a sort of ‘magnifying glass’ be applied, in order to capture those sense variations which select specific verb collocations

Does homeownership partly explain low participation in supplementary pension schemes?

We used nine waves of the Bank of Italy’s Survey on Household income and Wealth (1995- 2012) to investigate a possible trade-off between homeownership and individual participation in a supplementary pension scheme. Italy lends itself to this type of investigation because the Italian public pension system has been heavily reformed in the period, providing in principle incentives for participation, and the homeownership rate is very high. The impact of homeownership is captured in two ways: by a dummy for being homeowner and by an index defined as the share of housing wealth over total wealth. Our results show that indeed, after controlling for a vast array of socio-economic characteristics and allowing for unobserved individual heterogeneity, both measures of homeownership are negatively associated with participation in supplementary pension schemes and that such an effect does not disappear even after the 2007 reform

Methods for Oxygen Determination in an NMR Bioreactor as a Surrogate Marker for Metabolomic Studies in Living Cell Cultures.

Nuclear magnetic resonance (NMR) approaches have been described as a powerful method for measuring oxygen in tissue cultures and body fluids by using relaxation time dependencies of substances on pO2. The present NMR study describes methods to longitudinally monitor global, in situ intracellular, and spatially resolved oxygen tension in culture media and 3D cell cultures using relaxation times of water without the need to use external sensors. 1H NMR measurements of water using a modified inversion recovery pulse scheme were employed for global, i.e., intra- and extracellular oxygen estimation in an NMR-bioreactor. The combination of 1H relaxation time T1 and diffusion measurements of water was employed for in situ cellular oxygen content determination. Spatially selective water relaxation time estimations were used for spatially resolved oxygen quantification along the NMR tube length. The inclusion in a study protocol of the presented techniques for oxygen quantification, as a surrogate marker of oxidative phosphorylation (OXPHOS), provides the possibility to measure mitochondrial respiration and metabolic changes simultaneously

Are patinas of the Mediterranean monuments really related to the rock substrate?

Patinas of different mineralogies and colours occur on most monuments of the Mediterranean area. The origin(s) of the latter have often been related to previous painting and/or protective treatment(s). Microbial and biogeochemical pathways of formation, however, have as well been proposed and discussed. Paintings and photographs make it clear that coatings of changing colours have covered famous monuments in the past 200 years. Also the presence of microbiota has been recorded already 150 years ago. The work recorded here was done on several buildings in the Mediterranean area. A wide variety of localities and rock substrates and the patinas on them have been studied. Among the rocks were marbles, limestones calcarenites, granites and sandstones. In all cases studied patinas, their thickness and colour as well as their mineralogical composition were related to the exposure type. The patinas usually are multi-layered, the individual layers characterized by different crystal size, texture, mineralogy, and colour. Orange to grey layers are characterized by calcium carbonates and oxalates with some phosphates admixed, while the dark grey to black layers are rather characterized by gypsum with some trapped air-borne particles in cases. The petrology and mineralogy of the patinas is practically identical in all cases. Many of the patinas are inhabited by a variated microflora causing pitting and exfoliation in cases of the patina. Sometimes the pitting is only seen in the patina, and it reaches also the bedrock. A relationship between patina formation, preservation, destruction and climatic changes over the past 200 years is derived from these findings

Emerging Insights on the Biological Impact of Extracellular Vesicle-Associated ncRNAs in Multiple Myeloma

Increasing evidence indicates that extracellular vesicles (EVs) released from both tumor cells and the cells of the bone marrow microenvironment contribute to the pathobiology of multiple myeloma (MM). Recent studies on the mechanisms by which EVs exert their biological activity have indicated that the non-coding RNA (ncRNA) cargo is key in mediating their effect on MM development and progression. In this review, we will first discuss the role of EV-associated ncRNAs in different aspects of MM pathobiology, including proliferation, angiogenesis, bone disease development, and drug resistance. Finally, since ncRNAs carried by MM vesicles have also emerged as a promising tool for early diagnosis and therapy response prediction, we will report evidence of their potential use as clinical biomarkers

Determination of Intra- and Extracellular Metabolic Adaptations of 3D Cell Cultures upon Challenges in Real-Time by NMR.

NMR flow devices provide longitudinal real-time quantitative metabolome characterisation of living cells. However, discrimination of intra- and extracellular contributions to the spectra represents a major challenge in metabolomic NMR studies. The present NMR study demonstrates the possibility to quantitatively measure both metabolic intracellular fingerprints and extracellular footprints on human control fibroblasts by using a commercially available flow tube system with a standard 5 mm NMR probe. We performed a comprehensive 3D cell culture system characterisation. Diffusion NMR was employed for intra- and extracellular metabolites separation. In addition, complementary extracellular footprints were determined. The implemented perfused NMR bioreactor system allowed the determination of 35 metabolites and intra- and extracellular separation of 19 metabolites based on diffusion rate differences. We show the reliability and sensitivity of NMR diffusion measurements to detect metabolite concentration changes in both intra- and extracellular compartments during perfusion with different selective culture media, and upon complex I inhibition with rotenone. We also demonstrate the sensitivity of extracellular footprints to determine metabolic variations at different flow rates. The current method is of potential use for the metabolomic characterisation of defect fibroblasts and for improving physiological comprehension

A phase II, randomized, single-blinded, placebo-controlled clinical trial on the efficacy of Curcumina and Calendula suppositories for the treatment of patients with chronic prostatitis/chronic pelvic pain syndrome type III

Objective: The management of chronic prostatitis/ chronic pelvic pain syndrome type III (CP/CPPS) has been always considered complex due to several biopsychological factors underling the disease. In this clinical study, we aimed to evaluate the efficacy of the treatment with Curcumin and Calendula extract in patients with CP/CPPS III. Material and methods: From June 2015 to January 2016 we enrolled 60 consecutive patients affected by CP/CPPS III in our institution. Patients between 20 and 50 year of age with symptoms of pelvic pain for 3 months or more before study, a total National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI) score ≥ 15 point and diagnosed with NIH category III. Patients were then allocated to receive placebo (Group A) or treatment (Group B). Treatment consisted of rectal suppositories of Curcumin extract 350 mg (95%) and Calendula extract 80 mg (1 suppository/die for 1 month). Patients of Group B received 1 suppository/die for 1 month of placebo. The primary endpoint of the study was the reduction of NIH-CPSI. The secondary outcomes were the change of peak flow, IIEF-5, VAS score and of premature ejaculation diagnostic tool (PEDT). Results: A total of 48 patients concluded the study protocol. The median age of the all cohort was 32.0 years, the median NIH-CPSI was 20.5, the median IIEF-5 was 18.5, the median PEDT was 11.0, the median VAS score was 7.5 and the median peak flow was 14.0. After 3 months of therapy in group A we observed a significant improvement of NIH-CPSI (-5.5; p < 0.01), IIEF-5 (+ 3.5; p < 0.01), PEDT (-6.5; p < 0.01), peak flow (+2.8; p < 0.01) and VAS (-6.5; p < 0.01) with significant differences over placebo group (all p-value significant). Conclusions: In this phase II clinical trial we showed the clinical efficacy of the treatment with Curcumin and Calendula in patients with CP/CPPS III. The benefits of this treatment could be related to the reduction of inflammatory cytokines and of inflammatory cells. These results should be confirmed in further studies with greater sample size