Image analysis procedure for studying Back-Diffusion phenomena from low-permeability layers in laboratory tests

In this study, the long-term tailing derived from the storage process of contaminants in low-permeability zones is investigated. The release from these areas in the groundwater can be considered a long-term source that often undermines remediation efforts. An Image Analysis technique is used to analyze the process and evaluate the concentrations of a tracer at different points of the test section. Furthermore, the diffusive flux from the low-permeability lenses is determined. To validate the proposed technique, the results are compared with samples, and the diffusive fluxes resulting from the low-permeability zones of the reconstructed aquifer are compared with a theoretical approach

Non-surgical treatment of hidradenitis suppurativa: the role of cryotherapy

Background. Hidradenitis suppurativa is a chronic and recurrent inflammatory disease with a great impact on a patient's quality of life, due to the painful involvement of very sensitive areas, such as the groin, mammary area, and genitals, with malodourous discharge. Multiple treatment options are available; however, no single treatment is effective for all patients, and usually, a combination of medical therapy with various surgical and physical procedures is provided. Cryotherapy is not a routine technique to treat HS, although usually available in the majority of medical clinics, and is cheaper than laser and surgical treatments. The aim of this study was to evaluate the effectiveness of cryotherapy on persistent HS nodules, to reduce the burden of local disease. Materials and methodsRetrospective observational study in all patients treated with liquid nitrogen cryotherapy for persistent nodules of hidradenitis suppurativa in the last 2 years, with at least 6 months of follow-up after the procedure. Disease severity was assessed with Hurley staging and sonographic staging according to SOS-HS (18 MHz probe, Esaote-MyLab (TM)). The results were scored with a 0-3 points scale, as complete remission (3), partial response (2 to 1), or no response (0), after one session of treatment. Local cleansing and antiseptic treatment after the procedure was the same as previously performed in each patient, not to influence recovery. ResultsIn total, 23 patients were included, with a total of 71 persistent nodules treated with a single cryotherapy session. The treatment has been effective in 63 out of 71 nodules treated (88.7%), and the patients attested that they recommend the treatment, the discomfort during recovery was minimal, and the management was not different from daily routine. Persistence was considered as having a failure rate (11.3% overall) and occurred in 7.5% of the nodules of the axillary region, 18.2% on the groin, and 11.2% for nodules of the gluteal region. ConclusionCryotherapy is a simple and effective procedure for the treatment of persistent nodules of HS not responding to medical therapy, and it is a valid alternative to local surgery or laser ablation

Experimental and Numerical Study of Biochar Fixed Bed Column for the Adsorption of Arsenic from Aqueous Solutions

Two laboratory tests were carried out to verify the suitability of an Italian commercial biochar as an adsorbing material. The chosen contaminant, considered dissolved in groundwater, was As. The circular economic concept demands the use of such waste material. Its use has been studied in recent years on several contaminants. The possibility of using an efficient material at low cost could help the use of low-impact technologies like permeable reactive barriers (PRBs). A numerical model was used to derive the kinetic constant for two of the most used isotherms. The results are aligned with others derived from the literature, but they also indicate that the use of a large amount of biochar does not improve the efficiency of the removal. The particular origin of the biochar, together with its grain size, causes a decrease in contact time required for the adsorption. Furthermore, it is possible that a strong local decrease in the hydraulic conductibility does not allow for a correct dispersion of the flow, thereby limiting its efficiency

Glucosylceramidase Mass and Subcellular Localization Are Modulated by Cholesterol in Niemann-Pick Disease Type C

Niemann-Pick disease type C (NPC) is characterized by the accumulation of cholesterol and sphingolipids in the late endosomal/lysosomal compartment. The mechanism by which the concentration of sphingolipids such as glucosylceramide is increased in this disease is poorly understood. We have found that, in NPC fibroblasts, the cholesterol storage affects the stability of glucosylceramidase (GCase), decreasing its mass and activity; a reduction of cholesterol raises the level of GCase to nearly normal values. GCase is activated and stabilized by saposin C (Sap C) and anionic phospholipids. Here we show by immunofluorescence microscopy that in normal fibroblasts, GCase, Sap C, and lysobisphosphatidic acid (LBPA), the most abundant anionic phospholipid in the endolysosomal system, reside in the same intracellular vesicular structures. In contrast, the colocalization of GCase, Sap C, and LBPA is markedly impaired in NPC fibroblasts but can be re-established by cholesterol depletion. These data show for the first time that the level of cholesterol modulates the interaction of GCase with its protein and lipid activators, namely Sap C and LBPA, regulating the GCase activity and stability

The secretion and maturation of prosaposin and procathepsin D are blocked in embryonic neural progenitor cells

The notion that prosaposin (Prosap) is likely involved in brain development and regeneration led us to explore its expression in stem/progenitor neural cells and its fate after cell differentiation. The expression of procathepsin-cathepsin D (proCath-Cath D), an endoprotease that plays an important role in the processing and sorting of Prosap, has been concomitantly examined. Our data evidenced that in embryonic human neural progenitor cells (eHNPCs) intact and high molecular weight intermediate forms of Prosap and intermediate forms of Cath D accumulated inside the cells, while the formation of saposins and mature Cath D was impaired. Furthermore, neither Prosap nor proCath D were secreted from eHNPCs. The block of the processing and secretion shared by Prosap and proCath D was overcome during the course of differentiation of eHNPCs into a mixed population of astrocytes and neuronal cells. Upon differentiation, large amounts of Prosap and proCath D were secreted from the cells, while saposins and mature Cath D were produced inside the cells. The dramatic accumulation of Prosap (an antiapoptotic factor) and reduction of mature Cath D (a proapoptotic factor) in the undifferentiated eHNPCs most likely play a role in the molecular mechanisms regulating the resistance to apoptotic signals of these cells and might represent a critically important issue in HNPCs biology. © 2008 Elsevier B.V. All rights reserved

Hyperactive HRAS dysregulates energetic metabolism in fibroblasts from patients with Costello syndrome via enhanced production of reactive oxidizing species

Germline-activating mutations in HRAS cause Costello syndrome (CS), a cancer prone multisystem disorder characterized by reduced postnatal growth. In CS, poor weight gain and growth are not caused by low caloric intake. Here, we show that constitutive plasma membrane translocation and activation of the GLUT4 glucose transporter, via reactive oxygen species-dependent AMP-activated protein kinase α and p38 hyperactivation, occurs in primary fibroblasts of CS patients, resulting in accelerated glycolysis and increased fatty acid synthesis and storage as lipid droplets. An accelerated autophagic flux was also identified as contributing to the increased energetic expenditure in CS. Concomitant inhibition of p38 and PI3K signaling by wortmannin was able to rescue both the dysregulated glucose intake and accelerated autophagic flux. Our findings provide a mechanistic link between upregulated HRAS function, defective growth and increased resting energetic expenditure in CS, and document that targeting p38 and PI3K signaling is able to revert this metabolic dysfunction.n

How future surgery will benefit from SARS-COV-2-related measures: a SPIGC survey conveying the perspective of Italian surgeons

COVID-19 negatively affected surgical activity, but the potential benefits resulting from adopted measures remain unclear. The aim of this study was to evaluate the change in surgical activity and potential benefit from COVID-19 measures in perspective of Italian surgeons on behalf of SPIGC. A nationwide online survey on surgical practice before, during, and after COVID-19 pandemic was conducted in March-April 2022 (NCT:05323851). Effects of COVID-19 hospital-related measures on surgical patients' management and personal professional development across surgical specialties were explored. Data on demographics, pre-operative/peri-operative/post-operative management, and professional development were collected. Outcomes were matched with the corresponding volume. Four hundred and seventy-three respondents were included in final analysis across 14 surgical specialties. Since SARS-CoV-2 pandemic, application of telematic consultations (4.1% vs. 21.6%; p < 0.0001) and diagnostic evaluations (16.4% vs. 42.2%; p < 0.0001) increased. Elective surgical activities significantly reduced and surgeons opted more frequently for conservative management with a possible indication for elective (26.3% vs. 35.7%; p < 0.0001) or urgent (20.4% vs. 38.5%; p < 0.0001) surgery. All new COVID-related measures are perceived to be maintained in the future. Surgeons' personal education online increased from 12.6% (pre-COVID) to 86.6% (post-COVID; p < 0.0001). Online educational activities are considered a beneficial effect from COVID pandemic (56.4%). COVID-19 had a great impact on surgical specialties, with significant reduction of operation volume. However, some forced changes turned out to be benefits. Isolation measures pushed the use of telemedicine and telemetric devices for outpatient practice and favored communication for educational purposes and surgeon-patient/family communication. From the Italian surgeons' perspective, COVID-related measures will continue to influence future surgical clinical practice

Il "modello Roma" dall'Italia all'Europa (1690-1789): arte, teatro, letteratura, architettura dalla fondazione dell'Arcadia alla fine dell'Antico Regime. Una mappatura culturale dei luoghi e degli eventi.

L'obiettivo principale della ricerca è la costruzione di una mappatura dei luoghi della cultura a Roma tra Sei e Settecento al fine di superare le contrastanti letture date da prospettive disciplinari diverse. L'apporto degli storici dell'arte e dell'architettura è fondamentale per dare rilievo alla ricchissima produzione letteraria, da inquadrare all'interno degli stessi progetti culturali che danno vita in questi decenni a un formidabile sviluppo delle strutture architettoniche (palazzi, ville, musei, biblioteche, ecc.) La ricerca è articolata in tre punti. Il primo punto, inquadramento della ricerca, si propone di rovesciare lo stereotipo della decadenza della Roma letteraria settecentesca e di far risaltare il valore aggiunto dato dal rapporto con l'antico (Roma come spazio di un Classicismo persistente). Il secondo punto prevede la definizione di un metodo che evidenzia le strette connessioni fra luoghii ed eventi. In una prima fase si individuano alcuni eventi di svolta per la cultura letteraria, artistica e architettonica romana sullo fondo della situazione storico-politica dalla fondazione dell'Arcadia al pontificato di Pio VI; in una seconda fase si svolgono per tutti gli ambiti disciplinari previsti ricerche d'archivio sui temi focalizzati, mentre nella terza si farà convergere il materiale raccolto nella costruzione di una mappatura tematica della città nel Settecento. Il terzo punto replicabile per qualunque luogo e per qualunque periodo storico, che valorizza però la specificità romana di questo secolo, permetterà la costruzione di un volume, quale primo data base di dati, studi, ricerca archivistiche e iconografiche. Grazie a un eventuale finanziamento europeo si potrà progettare un'applicazione digitale che correli i luoghi della cultura con le manifestazioni artistiche, teatrali, letterarie attraverso schede testuali e apparati iconografici. Ciò al fine di valorizzare la conoscenza e la divulgazione del patrimonio dei nostri beni culturali

Metabolically labeled exosomes for biogenesis and functional studies

Introduction: Exosomes are small Extracellular Vesicles (sEV) formed by an endosomal route by inward budding of the lateendosome/multivesicular body (MVB) membrane. Despite in recent years much progress has been made to better define sEVcomposition and biogenesis pathways, their small size and heterogeneity pose challenges to find new reliable labelling strategies toidentify specific exosome populations. We developed an innovative methodology to metabolically label fluorescent sEV throughthe use of a fluorescent lipid (BODIPY C16) that is readily internalized by cells and is transformed into phospholipids which willform part of the lipid bilayer of the secreted vesicles.Methods: Fluorescent sEV secreted in the conditioned media of melanoma cells pulsed with BODIPY FL C16 were purifiedby differential ultracentrifugation, quantified by Flow Cytometry (FC) and Nanoparticle Tracking Analysis (NTA), sorted byFluorescence Activated Cell Sorting (FACS) and further characterized by density gradient separation and Western Blot analysisfor typical sEVs markers. Colocalization studies were performed by confocal microscopy and electron microscopy.Results: Confocal images showed colocalization of BODIPY lipids with lipid transformation sites such as ER and mitochondriaand with specific markers of late endosomes/MVB or other organelles (tetraspanins, Golgi markers, lysosomes) but not with the of ABSTRACTplasma membrane. Secretion of fluorescent sEV (Bodipy sEV) was followed over time showing an early release of Bodipy sEVinto the extracellular medium with a constant ratio of Bodipy sEV/total EVs, as determined by NTA, up to 6 hours. Bodipy sEVsecreted in the conditioned media purified by differential ultracentrifugation were separated by density gradient fractionation.Fractions analysed by FC displayed a single low density peak at 1,08-1,09 g/ml that is detergent sensitive demonstrating thatfluorescent particles are indeed lipid vesicles and contain tetraspanins (CD63, CD81 and CD9), syntenin and ESCRT componentswhen analysed by Western Blot. Electron microscopy analysis of ultracentrifuged and sorted Bodipy sEV showed that BodipysEVhavethetypicalshapeandsize(about80nm)ofasubpopulationofsEVoftenreferredtoassmallexosomes(Exo-S).Finally,colocalization studies of single Bodipy sEV with tetraspanins fluorescent antibodies showed colocalization of Bodipy sEV withCD63, CD81 and CD9.Summary/Conclusion: Taken together these results show a very specific and effective labelling of a discrete sEV subpopulationthat can be further exploited for biogenesis, internalization and functional studies. This work was supported by the Italian Ministry of Health (grant RF-2019-12369719