Adult Human Vascular Smooth Muscle Cells on 3D Silk Fibroin Nonwovens Release Exosomes Enriched in Angiogenic and Growth-Promoting Factors

Background. Our earlier works showed the quick vascularization of mouse skin grafted Bombyx mori 3D silk fibroin nonwoven scaffolds (3D-SFnws) and the release of exosomes enriched in angiogenic/growth factors (AGFs) from in vitro 3D-SFnws-stuck human dermal fibroblasts (HDFs). Here, we explored whether coronary artery adult human smooth muscle cells (AHSMCs) also release AGFs-enriched exosomes when cultured on 3D-SFnws in vitro. Methods. Media with exosome-depleted FBS served for AHSMCs and human endothelial cells (HECs) cultures on 3D-SFnws or polystyrene. Biochemical methods and double-antibody arrays assessed cell growth, metabolism, and intracellular TGF-β and NF-κB signalling pathways activation. AGFs conveyed by CD9+/CD81+ exosomes released from AHSMCs were double-antibody array analysed and their angiogenic power evaluated on HECs in vitro. Results. AHSMCs grew and consumed D-glucose more intensely and showed a stronger phosphorylation/activation of TAK-1, SMAD-1/-2/-4/-5, ATF-2, c-JUN, ATM, CREB, and an IκBα phosphorylation/inactivation on SFnws vs. polystyrene, consistent overall with a proliferative/secretory phenotype. SFnws-stuck AHSMCs also released exosomes richer in IL-1α/-2/-4/-6/-8; bFGF; GM-CSF; and GRO-α/-β/-γ, which strongly stimulated HECs’ growth, migration, and tubes/nodes assembly in vitro. Conclusions. Altogether, the intensified AGFs exosomal release from 3D-SFnws-attached AHSMCs and HDFs could advance grafts’ colonization, vascularization, and take in vivo—noteworthy assets for prospective clinical applications

Opposite effects of cell differentiation and apoptosis on Ap3A/Ap4A ratio in human cell cultures.

The biological role of diadenosine oligophosphates (DAOP) remains obscure in spite of numerous attempts to solve this enigma. It is known that Ap3A contrary to Ap4A accumulates in human cultured cells treated with interferons (IFNs) alpha or gamma. Since IFNs are considered as antiproliferative regulators, we assumed that different cell status may be associated with varying intracellular levels of DAOP. Promyelocytic human cell line HL60 induced by phorbol ester (TPA) to differentiate to macrophage-like cells in culture exhibits a profound loss of proliferative potential. Here we have shown a 4-5-fold increase in Ap3A concentration in HL60 cells induced by TPA, similar to the effect of IFN, while the Ap4A concentration remained unchanged. On the contrary, in cells undergoing apoptosis induced by VP16, a topoisomerase II inhibitor, the Ap3A concentration considerably decreased, while the Ap4A concentration increased. These findings combined with earlier results suggest an involvement of the Ap3A/Ap4A ratio in signal transduction pathways controlling the cell status

Biomolecular characterization of Fusarium poae strains isolated from durum wheat in central Italy.

Fusarium Head Blight (FHB) is a worldwide disease affecting wheat, barley and other grains, reducing kernel weight and grain yield; infected seeds may contain a large number of mycotoxins, including trichothe- cenes of type A and B. These compounds have already been associated with human and animal toxicoses. Most common species causing the disease are F. graminearum, F. culmorum and F. avenaceum, but in the last few years a gradual increase in incidence of another species, F. poae, has been reported. In general terms, F. poae is a relatively weak pathogen, but its contribute to the increase of mycotoxins level still has to be clarified. Durum wheat is widely cultivated in the central part of Italy, however the effective incidence of F. poae in this area still has to be investigated. In order to monitor Fusarium risk, we collected dozens of F. poae strains on seeds and glumes of durum wheat coming from some of the most important cultivated areas of Central Italy. Every isolate was identified both by microscope observation and by PCR assay with the primer pair Fp82 F/R. Strains were therefore subjected to a more accurate molecular characterization by Translation Elongation Factor 1-alpha (TEF-1α) gene sequencing.

Caratterizzazione biomolecolare di ceppi di Fusarium spp. isolati da seme di Eruca sativa e Diplotaxis spp.

Recenti infezioni a carico di rucola coltivata e selvatica in Italia sono state causate da Fusarium oxysporum f. sp. raphani. Considerando che i miceti riferibili alla specie F. oxysporum sono agenti di deperimenti o marciumi radicali su centinaia di ospiti vegetali, distinti sulla base di risposte ospite specifiche in formae speciales e che la specie è ubiquitaria e cosmopolita, risulta difficile effettuare una identificazione tempestiva e certa con il solo approccio morfologico. L’esatta identificazione degli agenti causali è punto cruciale nella valutazione della sanità delle sementi e nella gestione di questa problematica. Nel presente lavoro sono stati analizzati 39 lotti di semente, di cui 31 di rucola selvatica e 8 di rucola coltivata, su substrato semiselettivo (Komada’s medium) per la ricerca di Fusarium spp. Da questi lotti sono stati reperiti 24 isolati, che, unitamente a 2 ceppi di riferimento, sono stati saggiati biologicamente tramite prove di patogenicità differenziali in ambiente controllato e caratterizzati molecolarmente sulla base dello studio dei geni ITS e EF-1. Nessun isolato da seme, analizzato in questa indagine, risulta appartenere a F. oxysporum f. sp. raphani e la maggioranza dei ceppi fungini isolati appartiene a Gibberella fujikuroi Complex

Determination of deoxynivalenol and nivalenol producing chemotypes of Fusarium graminearum isolated from durum wheat in different Italian regions.

Durum wheat production in Italy is economically of great importance. Fusarium graminearum is the main fusarium head blight (FHB) causal agent in wheat, reducing both yield and grain quality. F. graminearum produces several mycotoxins and, among trichothecenes, deoxynivalenol (DON) and nivalenol (NIV) are the most studied for their toxicity towards humans and animals. DON-producing isolates can be further distin- guished on the basis of the predominant acetyl-DON derivative in 3-acetyldeoxynivalenol (3-ADON) or 15- acetyldeoxynivalenol (15-ADON). In order to evaluate possible mycotoxin contamination risks in food, it is very important to know which chemotype is the prevalent in a F. graminearum population. F. graminearum sensu stricto strains were collected from symptomatic durum wheat heads and grains of several naturally infected fields located mostly in Emilia – Romagna, The Marche, Lazio, Tuscany and Umbria. A multiplex PCR in the region of genes Tri12, located in the terminal gene cluster of trichothecenes, was used to characterize 187 single-spore isolates of F. graminearum as NIV, 3-ADON and 15-ADON chemotypes. All the three chemotypes were present in the F. graminearum population studied. The most frequent chemotype was 15-ADON (83.4%), followed by 3-ADON (10.7%) and NIV (5.9%). NIV-producing isolates were found only in Emilia-Romagna (3.5%), Umbria (33.3%) and The Marche (5.7%)