The Autophagy-Related Organelle Autophagoproteasome Is Suppressed within Ischemic Penumbra

The peri-infarct region, which surrounds the irreversible ischemic stroke area is named ischemic penumbra. This term emphasizes the borderline conditions for neurons placed within such a critical region. Area penumbra separates the ischemic core, where frank cell loss occurs, from the surrounding healthy brain tissue. Within such a brain region, nervous matter, and mostly neurons are impaired concerning metabolic conditions. The classic biochemical marker, which reliably marks area penumbra is the over-expression of the heat shock protein 70 (HSP70). However, other proteins related to cell clearing pathways are modified within area penumbra. Among these, autophagy proteins like LC3 increase in a way, which recapitulates Hsp70. In contrast, components, such as P20S, markedly decrease. Despite apparent discrepancies, the present study indicates remarkable overlapping between LC3 and P20S redistribution within area penumbra. In fact, the amount of both proteins is markedly reduced within vacuoles. Specifically, a massive loss of LC3 + P20S immuno-positive vacuoles (autophagoproteasomes) is reported here. This represents the most relevant sub-cellular alteration here described in cell clearing pathways within area penumbra. The functional significance of these findings remains to be determined and it will take a novel experimental stream to decipher the fine-tuning of such a phenomenon.     

Thermal behaviour assessment of a novel vertical greenery module system: first results of a long-term monitoring campaign in an outdoor test cell

Vertical greenery modular systems (VGMSs) are an increasingly widespread building envelope solution aimed at improving the aesthetical quality of both new and existing façades, contemporarily achieving high energy efficiency performance. Within a research project, a new prototype of VGMS was developed, designed and tested. An experimental monitoring campaign was carried out on a test cell located in Turin (northern Italy), aimed at assessing both biometric parameters and energy-related issues. Two different types of growing media and two plant species, Lonicera nitida L. and Bergenia cordifolia L., have been tested on a south-facing lightweight wall. Results have been compared to the same wall without VGMS and plaster finished, in order to characterise the thermal insulation effectiveness in the winter period and the heat gain reduction in the summer period. Measured equivalent thermal transmittance values of the green modular system showed a 40 % reduction, when compared to the plastered wall, thus noticeably impacting on the energy crossing the façade during the heating season. Benefits of the VGMS are measured also during the summer season, when the presence of vegetation lowers the outdoor surface temperatures of the wall up to 23 °C compared to the plastered finishing, with a positive effect on outdoor comfort and urban heat island mitigation. Nevertheless, as far as the entering energies are concerned, not significant reduction was observed for VGMS, compared to the reference plastered wall, since the green coverage acts as a thermal buffer and solar radiation is stored and slowly released to the indoor environment.     

Drug-Induced Lysosomal Impairment Is Associated with the Release of Extracellular Vesicles Carrying Autophagy Markers

Amiodarone is a cationic amphiphilic drug used as an antiarrhythmic agent. It induces phospholipidosis, i.e., the accumulation of phospholipids within organelles of the endosomal–lysosomal system. Extracellular vesicles (EVs) are membrane-enclosed structures released by any type of cell and retrieved in every fluid of the body. EVs have been initially identified as a system to dispose cell waste, but they are also considered to be an additional manner to transmit intercellular signals. To understand the role of EVs in drug-induced phospholipidosis, we investigated EVs release in amiodarone-treated HEK-293 cells engineered to produce fluorescently labelled EVs. We observed that amiodarone induces the release of a higher number of EVs, mostly of a large/medium size. EVs released upon amiodarone treatment do not display significant morphological changes or altered size distribution, but they show a dose-dependent increase in autophagy associated markers, indicating a higher release of EVs with an autophagosome-like phenotype. Large/medium EVs also show a higher content of phospholipids. Drugs inducing lysosomal impairment such as chloroquine and bafilomycin A1 similarly prompt a higher release of EVs enriched in autophagy markers. This result suggests a mechanism associated with amiodarone-induced lysosomal impairment more than a connection with the accumulation of specific undigested substrates. Moreover, the implementation of the lysosomal function by overexpressing TFEB, a master gene regulator of lysosomal biogenesis, prevents the amiodarone-induced release of EVs, suggesting that this could be a feasible target to attenuate drug-induced abnormalities.     

Production of hydrogen peroxide is responsible for the induction of apoptosis by hydroxytyrosol on HL60 cells

Hydroxytyrosol [3,4‐dihydroxyphenylethanol (3,4‐DHPEA)], a phenolic compound found exclusively in olive oil, exerts growth‐suppressive and pro‐apoptotic effects on different cancer cells. Although some molecular mechanisms involved in the pro‐apoptotic activity of 3,4‐DHPEA have been proposed, the initial stress signals responsible of this phenomenon are not known. Our aim was to assess the involvement of reactive oxygen species as mediators of apoptosis induced by 3,4‐DHPEA on HL60 cells. Apoptosis was determined by analyzing the nuclear fragmentation by both fluorescence microscopy and flow cytometry. The externalization of phosphatidylserine was evidenced using an Annexin V‐FITC kit. The concentration of H₂O₂ in the culture medium was measured by the ferrous ion oxidation‐xylenol orange method. The pro‐apoptotic effect of 3,4‐DHPEA (100 μM) was prevented by N‐acetyl‐cysteine, ascorbate, and α‐tocopherol. Catalase suppressed the 3,4‐DHPEA‐induced apoptosis, while the Fe(II)‐chelating reagent o‐phenantroline showed no effect, suggesting the involvement of H₂O₂ but not of OH^?. Indeed, 3,4‐DHPEA caused accumulation of H₂O₂ in the culture medium. Tyrosol (p‐hydroxyphenylethanol) and caffeic acid, compounds structurally similar to 3,4‐DHPEA but not able to generate H₂O₂, did not induce an appreciable apoptotic effect. This is the first study demonstrating that apoptosis induction by 3,4‐DHPEA is mediated by the extracellular production of H₂O₂.     

Identification of natural epimeric flavanone glycosides by NMR spectroscopy

Recently advanced analytical technology has provided evidence of the existence of stereoisomers of many natural products. Particularly, flavanones which might have two different configurations at C-2 exist in many food additives, e.g., citrus fruits. In this study, the possible stereoisomers of flavanone glycosides were identified by NMR spectroscopy. Based on NMR spectra of common flavanone glycosides such as naringin, hesperidin, and neohesperidin, the two existing diastereomeric forms of the molecules could clearly be distinguished. The ¹H NMR resonances of two diastereomers of each flavanone glycosides investigated in this study were fully assigned with the assistance of diverse 2D NMR spectroscopy methods.     

Sample treatment considerations in the analysis of organic colorants by surface-enhanced Raman scattering

The introduction of surface-enhanced Raman spectroscopy (SERS) in the field of cultural heritage has significantly improved the analysis of the organic dyes and their complexes that have been used as textile dyes and pigments in paintings and other polychrome works of art since antiquity. Over the last five years, a number of different procedures have been developed by various research groups. In this Article, we evaluate the effect of pretreating samples by exposing them to hydrofluoric acid (HF) vapor prior to SERS analysis, a step designed to hydrolyze the dye-metal complexes and increase analyte adsorption on the nanosized metallic support, thus enhancing the SERS signal. Materials studied include pure colorants, commercial lake pigments, and fibers from dyed textiles, as well as actual aged samples, such as microscopic fragments of lakes on paper and ancient pigments and glazes from several works of art, covering a wide range of time, from the second century B.C. to the early 20th century. In each case, SERS spectra obtained with or without HF hydrolysis were critically evaluated. The pretreatment with HF vapor resulted in faster analysis and increased sensitivity in most cases, with the exception of dyed silk fibers, where silk protein hydrolyzates were found to interfere with SERS analysis. As a final point, a two-step procedure including SERS on untreated and treated samples is proposed as a standard approach: by analyzing a sample first without hydrolysis, and then, following removal of the colloid, upon HF treatment, the best and most reliable results for a great number of dyes and substrates are assured.     

Combined hydrogen production and power generation from aluminum combustion with water: Analysis of the concept

In this paper the combined production of hydrogen and power based on the aluminum combustion with water is investigated. Furthermore, a concept system is proposed that is potentially able to provide pressurized hydrogen and high temperature steam along with heat and work at the crankshaft. The system demonstrates high energy conversion efficiency, and it fully complies with environment sustainability requirements.     

Transferable Antibiotic Resistances in Marketed Edible Grasshoppers (Locusta migratoria migratorioides)

Grasshoppers are the most commonly eaten insects by humans worldwide, as they are rich in proteins and micronutrients. This study aimed to assess the occurrence of transferable antibiotic resistance genes in commercialized edible grasshoppers. To this end, the prevalence of 12 selected genes [aac(6’)‐Ie aph(2″)‐Ia, blaZ, erm(A), erm(B), erm(C), mecA, tet(M), tet(O), tet(S), tet(K), vanA, vanB] coding for resistance to antibiotics conventionally used in clinical practice was determined. The majority of samples were positive for tet(M) (70.0%), tet(K) (83.3%) and blaZ (83.3%). A low percentage of samples were positive for erm(B) (16.7%), erm(C) (26.7%), and aac(6’)‐Ie aph(2″)‐Ia (13.3%), whereas no samples were positive for erm(A), vanA, vanB, tet(O), and mecA. Cluster analysis identified 4 main clusters, allowing a separation of samples on the basis of their country of origin.