Exosome-mediated transfer of miR-222 is sufficient to increase tumor malignancy in melanoma

BACKGROUND: Growing evidence is showing that metastatic cell populations are able to transfer their characteristics to less malignant cells. Exosomes (EXOs) are membrane vesicles of endocytic origin able to convey their cargo of mRNAs, microRNAs (miRs), proteins and lipids from donors to proximal as well as distant acceptor cells. Our previous results indicated that miR-221&222 are key factors for melanoma development and dissemination. The aim of this study was to verify whether the tumorigenic properties associated with miR-222 overexpression can be also propagated by miR-222-containing EXOs. METHODS: EXOs were isolated by UltraCentrifugation or Exoquick-TC(®) methods. Preparations of melanoma-derived vesicles were characterized by using the Nanosight™ technology and the expression of exosome markers analyzed by western blot. The expression levels of endogenous and exosomal miRNAs were examined by real time PCR. Confocal microscopy was used to evaluate transfer and uptake of microvesicles from donor to recipient cells. The functional significance of exosomal miR-222 was estimated by analyzing the vessel-like process formation, as well as cell cycle rates, invasive and chemotactic capabilities. RESULTS: Besides microvesicle marker characterization, we evidenced that miR-222 exosomal expression mostly reflected its abundance in the cells of origin, correctly paralleled by repression of its target genes, such as p27Kip1, and induction of the PI3K/AKT pathway, thus confirming its functional implication in cancer. The possible differential significance of PI3K/AKT blockade was assessed by using the BKM120 inhibitor in miR-222-transduced cell lines. In addition, in vitro cultures showed that vesicles released by miR-222-overexpressing cells were able to transfer miR-222-dependent malignancy when taken-up by recipient primary melanomas. Results were confirmed by antagomiR-221&222 treatments and by functional observations after internalization of EXOs devoid of these miRs

3D-printed, home-made, UV-LED photoreactor as a simple and economic tool to perform photochemical reactions in high school laboratories

Abstract In the paper we present the simple manufacturing of an easy and economical UV-A photoreactor using a desktop 3D printer and its application in chemical transformations. PLA (polylactic acid) was used as inexpensive and not toxic polymer for the 3D printing process, while commercially available decorative ultraviolet LEDs (UV-A) have been employed as a light source. With this device, the photoreduction of benzophenone was performed in high yield and short times, compatible with the duration of a typical laboratory experiment in a high school program

SCD5 restored expression favors differentiation and epithelial-mesenchymal reversion in advanced melanoma

Our previous data supported a role for the Stearoyl-CoA desaturase (SCD5) in protection against malignancy, whereby it appears to functionally modify tumor stroma impairing tumor spread. SCD5 is significantly expressed in primary melanoma, but becomes barely detectable at tumor advanced stages. Looking for the regulatory mechanisms underlying SCD5 reduced expression during melanoma progression, we demonstrated a significantly lower stability of SCD5 protein as well as the direct targeting of SCD5 mRNA by the oncogenic miR-221 & 222 in metastatic cell lines. Moreover, our results indicated the existence of a negative feedback loop between SCD5 and miR-221 & 222, in good agreement with their opposite functions. Also, we showed how SCD5 re-expression and the direct supplementation of its main product oleic acid (OA) can drive advanced melanoma cell lines toward differentiation and reversion of the epithelial-mesenchymal (EMT)-like process, eventually inducing a less malignant phenotype. Indeed, SCD5 re-established the sensitivity to all-trans retinoic acid in A375M metastatic melanoma, associated with increased levels of Tyrosinase, melanin production and reduced proliferation. As evidenced by the correct modulation of some key transcription factors, SCD5 managed by favoring a partial mesenchymal-to-epithelial (MET) transition in in vitro studies. Interestingly, a more complete MET, including E-cadherin re-expression correctly localized at cell membranes, was obtained in in vivo xenograft models, thus indicating the requirement of direct contacts between tumor cells and the surrounding microenvironment as well as the presence of some essential factors for SCD5 complete function

News from the San Antonio Breast Cancer Symposium 2022

The 45th San Antonio Breast Cancer Symposium, held December 6–10 in San Antonio, Texas is the largest breast cancer conference and this year saw the participation of nearly 10,000 clinicians, researchers, and patient advocates, in person. Scientists shared many important new findings that are going to change the clinical practice in the near future. Here, we will present the most important news with a group of Italian colleagues and we will discuss how these results will impact the management of breast cancer

Efficient District Heating in a Decarbonisation Perspective: A Case Study in Italy

The European and national regulations in the decarbonisation path towards 2050 promote district heating in achieving the goals of efficiency, energy sustainability, use of renewables, and reduction of fossil fuel use. Improved management and optimisation, use of RES, and waste heat/cold sources decrease the overall demand for primary energy, a condition that is further supported by building renovations and new construction of under (almost) zero energy buildings, with a foreseeable decrease in the temperature of domestic heating systems. Models for the simulation of efficient thermal networks were implemented and described in this paper, together with results from a real case study in Italy, i.e., University Campus of Parma. Activities include the creation and validation of calculation codes and specific models in the Modelica language (Dymola software), aimed at investigating stationary regimes and dynamic behaviour as well. An indirect heat exchange substation was coupled with a resistive-capacitive model, which describes the building behaviour and the thermal exchanges by the use of thermos-physical parameters. To optimise indoor comfort conditions and minimise consumption, dynamic simulations were carried out for different operating sets: modulating the supply temperature in the plant depending on external conditions (Scenario 4) decreases the supplied thermal energy (-2.34%) and heat losses (-8.91%), even if a lower temperature level results in higher electricity consumption for pumping (+12.96%), the total energy consumption is reduced by 1.41%. A simulation of the entire heating season was performed for the optimised scenario, combining benefits from turning off the supply in the case of no thermal demand (Scenario 3) and from the modulation of the supply temperature (Scenario 4), resulting in lower energy consumption (the thermal energy supplied by the power plant -3.54%, pumping +7.76%), operating costs (-2.40), and emissions (-3.02%). The energy balance ex-ante and ex-post deep renovation in a single user was then assessed, showing how lowering the network operating temperature at 55 degrees C decreases the supplied thermal energy (-22.38%) and heat losses (-22.11%) with a slightly higher pumping consumption (+3.28%), while maintaining good comfort conditions. These promising results are useful for evaluating the application of low-temperature operations to the existing district heating networks, especially for large interventions of building renovation, and confirm their potential contribution to the energy efficiency targets

A microfluidic method for passive trapping of sperms in microstructures

Sperm motility is a prerequisite for male fertility. Enhancing the concentration of motile sperms in assisted reproductive technologies - for human and animal reproduction - is typically achieved through aggressive methods such as centrifugation. Here we propose a passive technique for the amplification of motile sperm concentration, with no externally imposed forces or flows. The technique is based upon the disparity between probability rates, for motile cells, of entering in and escaping from complex structures. The effectiveness of the technique is demonstrated in microfluidic experiments with microstructured devices, comparing the trapping power in different geometries. In these micro-traps we observe an enhancement of cells concentration close to 10, with a contrast between motile and non-motile increased by a similar factor. Simulations of suitable interacting model sperms in realistic geometries reproduce quantitatively the experimental results, extend the range of observations and highlight the ingredients that are key to optimal trap design

Pratiche per l’insegnamento adattivo nelle discipline STEAM: il Progetto T.E.S.T.

Il superamento della pandemia da Covid-19 ha impresso una spinta propulsiva per ripensare i modelli didattici dell’insegnamento nelle STEAM, orientando la ricerca scientifica verso la costruzione di repertori metodologici di tipo learner-centered e inquiry-based, in grado di valorizzare l’apprendimento attraverso l’esperienza in contesti ad alto tasso di tecnologizzazione. A fronte di questo scenario, l’articolo presenta i risultati del primo segmento qualitativo-esplorativo del progetto di ricerca biennale T.E.S.T. - Technologies for STEAM Teaching. Delinea l’impianto metodologico del progetto, che ha consentito la costruzione di un repertorio metodologico per l’insegnamento adattivo e accessibile nelle STEAM, in fase di sperimentazione nelle otto università partner del progetto. Il focus verte sulle caratteristiche del profilo emergente del “docente adattivo” e sulle dimensioni progettuali dell’insegnamento adattivo nelle discipline STEAM (Barana et al. 2019; Hardy et al., 2019)

Diel observations on populations of Dinophysis and Alexandrium spp. (Dinophyceae) from the NW Adriatic Sea

Quanti-qualitative changes in Dinophysis and Alexatidrium populations from the Gulf of Venice NW Adriatic Sea - were studied on a short time scale (44 h cycles). Under hydrographic conditions of marked thermohaline stratification (July 1995), D. sacculus populations assembled in the surface waters (12 24 yt isopycnals). Division processes took place mostly in the early hours of the morning and at 14:00 h, 4 to 10 % of the population consisted of morphotypes of reduced size (possibly gametes resulting from successive cell divisions). Similar percentages were also observed in October for D. caudata, with a maximum division between 06:00 h and 10:00 h. During this month, coexisting populations of Alexandrium cf. fundyense and A. cf. tamarense displayed rhythmic changes in cell density that slightly differed from those exhibited in July by A. pseudogonyaulax, suggesting a circadian control of the cell cycle rather than a response to hydrographic changes