An optimized procedure for preparation of conditioned medium from Wharton’s jelly mesenchymal stromal cells isolated from umbilical cord

Cell-free therapy based on conditioned medium derived from mesenchymal stromal cells (MSCs) has gained attention in the field of protective and regenerative medicine. However, the exact composition and properties of MSC-derived conditioned media can vary greatly depending on multiple parameters, which hamper standardization. In this study, we have optimized a procedure for preparation of conditioned medium starting from efficient isolation, propagation and characterization of MSCs from human umbilical cord, using a culture medium supplemented with human platelet lysate as an alternative source to fetal bovine serum. Our procedure successfully maximizes the yield of viable MSCs that maintain canonical key features. Importantly, under these conditions, the compositional profile and biological effects elicited by the conditioned medium preparations derived from these MSC populations do not depend on donor individuality. Moreover, approximately 120 L of conditioned medium could be obtained from a single umbilical cord, which provides a suitable framework to produce industrial amounts of toxic-free conditioned medium with predictable composition

Geo-hazards of the San Vito peninsula offshore (southwestern Tyrrhenian Sea)

In this paper we present geomorphological hazard mapping of the San Vito Peninsula offshore (Sicilian margin of Tyrrhenian Sea), characterised by a very narrow continental shelf and a very dipping, tectonically active continental slope, no far from a seismogenic belt. The data set consists of morpho-bathymetric models achieved by means of multibeam survey and scattered high resolution seismic profiles. The morpho-bathymetric study highlighted some potential sites of geomorphological hazards, the most representative of which are: (i) two canyons developing from the upper slope off San Vito Cape down to the Erice basin; (ii) a dense network of small incisions across the upper slope; (iii) a very active landslide, developed onland and extending in the continental shelf close to the Scopello village. The geomorphological features that can generate submarine hazard are represented by: unstable sediment packages into canyon heads or along the upper slope; submarine canyons with regressive erosion; coastal landslides

Apelin-13 Increases Functional Connexin-43 through Autophagy Inhibition via AKT/mTOR Pathway in the Non-Myocytic Cell Population of the Heart

Studies have shown a link between the downregulation of connexin 43 (Cx43), the predominant isoform in cardiac gap junctions, and high susceptibility to cardiac arrhythmias and cardiomyocyte death. Non-myocytic cells (NMCs), the most abundant component of the heart, exert multiple cardiac functions and represent an important therapeutic target for diseased cardiac tissue. A few studies have investigated the effect of Apelin-13, an endogenous peptide with a key role in various cardiovascular functions, on Cx43 expression in cardiomyocytes. However, it remained unknown whether Apelin-13 influences Cx43 expression in NMCs. Here, we found that in NMCs, Cx43 protein expression increased after Apelin-13 treatment (100 nM for 48 h). Furthermore, dye transfer assays proved that Apelin-13-treated NMCs had a greater ability to communicate with surrounding cardiomyocytes, and this effect was abrogated by carbenoxolone, a gap junction inhibitor. Interestingly, we showed that Apelin-13 increased Cx43 through autophagy inhibition, as proved by the upregulation of p62 and LC3I, acting as 3-MA, a well-known autophagy inhibitor. In addition, Apelin-13-induced AKT and mTOR phosphorylation was abolished by LY294002 and rapamycin inhibitors resulting in Cx43 increased suppression. These results open the possibility of targeting gap junctions in NMCs with Apelin-13 as an exciting therapeutic approach with great potential

Viscoelastic bearings with fractional constitutive law for fractional tuned mass dampers

The paper aims at studying the effects of the inherent fractional constitutive law of viscoelastic bearings used as devices for tuned mass dampers. First, the proper constitutive law of the viscoelastic supports is determined by the local constitutive law. Then, the characteristic force-displacement relationship at the top of the bearing is found. Taking advantage of the whole bearing constitutive laws, the tuning of the mass damper is proposed by defining the damped fractional frequency, which is analogous to the classical damped frequency. The effectiveness of the optimal tuning procedure is validated by a numerical application on a system subjected to a Gaussian white noise

Molecular mechanisms of mitotane action in adrenocortical cancer based on in vitro studies

SIMPLE SUMMARY: Mitotane is the only approved drug for the treatment of advanced adrenocortical carcinoma and for postoperative adjuvant therapy. It is known that mitotane destroys the adrenal cortex impairing steroidogenesis, although its exact molecular mechanism is still unclear. However, confounding factors affecting in vitro experiments could reduce the relevance of the studies. In this review, we explore in vitro studies on mitotane effects, highlighting how different experimental conditions might contribute to the controversial findings. On this basis, it may be necessary to re-evaluate the experiments taking into account their potential confounding factors such as cell strains, culture serum, lipoprotein concentration, and culture passages, which could hide important molecular results. As a consequence, the identification of novel pharmacological molecular pathways might be used in the future to implement personalized therapy, maximizing the benefit of mitotane treatment while minimizing its toxicity. ABSTRACT: Mitotane is the only approved drug for the treatment of advanced adrenocortical carcinoma and is increasingly used for postoperative adjuvant therapy. Mitotane action involves the deregulation of cytochromes P450 enzymes, depolarization of mitochondrial membranes, and accumulation of free cholesterol, leading to cell death. Although it is known that mitotane destroys the adrenal cortex and impairs steroidogenesis, its exact mechanism of action is still unclear. The most used cell models are H295-derived cell strains and SW13 cell lines. The diverging results obtained in presumably identical cell lines highlight the need for a stable in vitro model and/or a standard methodology to perform experiments on H295 strains. The presence of several enzymatic targets responsive to mitotane in mitochondria and mitochondria-associated membranes causes progressive alteration in mitochondrial structure when cells were exposed to mitotane. Confounding factors of culture affecting in vitro experiments could reduce the significance of any molecular mechanism identified in vitro. To ensure experimental reproducibility, particular care should be taken in the choice of culture conditions: aspects such as cell strains, culture serum, lipoproteins concentration, and culture passages should be carefully considered and explicated in the presentation of results. We aimed to review in vitro studies on mitotane effects, highlighting how different experimental conditions might contribute to the controversial findings. If the concerns pointed out in this review will be overcome, the new insights into mitotane mechanism of action observed in-vitro could allow the identification of novel pharmacological molecular pathways to be used to implement personalized therapy

Submarine Geomorphology and Sedimentary Features around the Egadi Islands (Western Mediterranean Sea)

The physiography, the geomorphological features and the sedimentary bedforms offshore Egadi Islands (Italy) have been illustrated and mapped through an integrated analysis of high-resolution bathymetric, seismo-acoustic and sedimentological data. The study area is characterized by a wide, up to 25 km, continental shelf which is separated by a NNW trending linear incision, the Marettimo Channel, along which several erosional and depositional features have been detected and mapped. Sedimentary prograding wedges were detected at water depths between 100-125 m along the shelf margin, which accumulated during sea level fall and lowstand stages of the last glacio-eustatic cycle (post- MIS 5.5). The study detected several slope breaks defining scarps across the continental shelf, which were interpreted as coastal cliffs that originated during the post-LGM eustatic sea level rise. Several fields of different types of sedimentary bedforms, including 2-D and 3-D hydraulic dunes and sorted bedforms, were found across the continental shelf, providing evidence of a high hydrodynamic regime affecting the seafloor. Further on, the study recognized erosive and depositional features related to bottom currents (contourites) in the Marettimo Channel. These findings provide a better understanding of the morpho-sedimentary evolution of the Egadi Islands offshore in the latest Quaternary. Moreover, they offer essential scientific support for effectively managing the most valuable priority habitats for conservation , such as Posidonia oceanica meadow and coralline algae bioconstructions (Coralligenous habitat)

Utility of laser microdissection and pressure catapulting in the diagnosis of non small cell lung cancer: preliminary data.

Background: There are controversies about the adequacy of tumor tissue sample on which the sequencing of molecular diagnosis could be performed to achieve the targeted-therapy on lung cancer. The aim of this study is to demonstrate the role of the Laser Microdissection Pressure Catapulting (LMPC) technique to obtain adequate tumor tissue sample for the molecular analysis of gene mutations in the target therapy of lung cancer. Findings: From a consecutive series of 24 patients with a diagnosis of locally-advanced or metastatic Non Small Cell Lung Cancer (NSCLC), we performed 29 diagnostic procedures using the system of LMPC, to obtain an homogeneous samples where it was possible to run the sequencing of the 4 most frequently mutated exons of Epidermal Growth Factor Receptor (EGFR) (exon 18, 19, 20, 21). Results: There were 14 males (58.3%) and 10 females (41.7%), with a mean age of 61 years old. Twenty one patients were affected by adenocarcinomas, 2 by squamous cell carcinomas and 1 by large cell carcinoma. We were able to obtain the sequencing on 26 out 29 samples (89,6%) for EGFR mutation. EGFR mutation rate in our population was 7,7%. In 5 samples, we found a polymorphism in exon 20 and one of them carried a mutation on exon 18 as well. In another sample we found the deletion of exon 19. On the other 20 samples we did not find any mutation. Conclusions: Our preliminary data suggest that the LMPC technique permits to obtain the tumor cells sample more homogeneous facilitating the application of biological molecular analysis for EGFR-gene mutation in a larger number of patients with NSCLC

Elevated miR-34a expression and altered transcriptional profile are associated with adverse electromechanical remodeling in the heart of male rats exposed to social stress

This study investigated epigenetic risk factors that may contribute to stress-related cardiac disease in a rodent model. Experiment 1 was designed to evaluate the expression of microRNA-34a (miR-34a), a known modulator of both stress responses and cardiac pathophysiology, in the heart of male adult rats exposed to a single or repeated episodes of social defeat stress. Moreover, RNA sequencing was conducted to identify transcriptomic profile changes in the heart of repeatedly stressed rats. Experiment 2 was designed to assess cardiac electromechanical changes induced by repeated social defeat stress that may predispose rats to cardiac dysfunction. Results indicated a larger cardiac miR-34a expression after repeated social defeat stress compared to a control condition. This molecular modification was associated with increased vulnerability to pharmacologically induced arrhythmias and signs of systolic left ventricular dysfunction. Gene expression analysis identified clusters of differentially expressed genes in the heart of repeatedly stressed rats that are mainly associated with morphological and functional properties of the mitochondria and may be directly regulated by miR-34a. These results suggest the presence of an association between miR-34a overexpression and signs of adverse electromechanical remodeling in the heart of rats exposed to repeated social defeat stress, and point to compromised mitochondria efficiency as a potential mediator of this link. This rat model may provide a useful tool for investigating the causal relationship between miR-34a expression, mitochondrial (dys)function, and cardiac alterations under stressful conditions, which could have important implications in the context of stress-related cardiac disease