Morphometric analysis of hepatocellular nodular lesions in HCV cirrhosis

Background and aims. We generated a computerized morphometric model to evaluate and quantify the morphological features in large regenerative nodules (LRN), high-grade dysplastic nodules (HGDN) and hepatocellular carcinoma (HCC). Methods. Sixteen LRN, 10 HGDN and 16 HCC in HCV-cirrhotic livers, were stained with H&E, smooth muscle actin, CD34, CD31 and reticulin to evaluate volume and surface fractions. Results. On H&E stains, the features most discriminatory between LRN, HGDN and HCC were volume fraction and number of hepatocytes nuclei in unit volume, and hepatocyte nuclear/cytoplasmic ratio. On immunohistochemistry, volume fractions of capillarised sinusoids, capillary units and isolated arteries were significantly different among all groups and highest in HCC; surface fraction of reticulin was markedly decreased in HCC. Conclusions. Our morphometric model is an objective method of quantification of the morphological changes of the nodular lesions and it could be applied in studies involving histological evaluation of the spectrum of nodular lesions arising in the cirrhotic liver

Effect of resveratrol on matrix metalloproteinase-2 (MMP-2) and secreted protein acidic and rich in cysteine (SPARC) on human cultured glioblastoma cells

INTRODUCTION: Glioblastoma is a highly malignant brain tumor with a high-invasive phenotype, so the prognosis is unfavorable, even in response to multidisciplinary treatment strategies. Obviously, therefore, a better therapeutic strategy is needed. Resveratrol has been reported to be one of the most potent chemopreventive agents inhibiting the cellular processes associated with tumor development, including initiation, promotion, and progression. MATERIALS AND METHODS: In this study we used RT-PCR, western blot and SDS-zymography to investigate the effect of resveratrol on the expression of genes and proteins involved in the extracellular matrix remodeling associated with tumor invasion in human cultured glioblastoma cells treated for 24, 48 and 72 h. We analyzed the expression of matrix metalloproteinase-2 (MMP-2), the main mediator of glioblastoma invasiveness, and the Secreted Protein Acidic and Rich in Cysteine (SPARC), involved in the regulation of cell-matrix interactions. RESULTS: Our results show a dose-related decrease of MMP-2 mRNA and protein levels 72 h after resveratrol treatment, and lower SPARC gene and protein expression 72 h after resveratrol treatment. This indicates that resveratrol may influence the two major factors in the ECM remodeling occurring with tumor invasion, suggesting it may have uses as a therapeutic agent for brain tumors

Tumor–Stroma Cross-Talk in Human Pancreatic Ductal Adenocarcinoma : A Focus on the Effect of the Extracellular Matrix on Tumor Cell Phenotype and Invasive Potential

Abstract: The extracellular matrix (ECM) in the tumor microenvironment modulates the cancer cell phenotype, especially in pancreatic ductal adenocarcinoma (PDAC), a tumor characterized by an intense desmoplastic reaction. Because the epithelial-to-mesenchymal transition (EMT), a process that provides cancer cells with a metastatic phenotype, plays an important role in PDAC progression, the authors aimed to explore in vitro the interactions between human PDAC cells and ECM components of the PDAC microenvironment, focusing on the expression of EMT markers and matrix metalloproteinases (MMPs) that are able to digest the basement membrane during tumor invasion. EMT markers and the invasive potential of HPAF-II, HPAC, and PL45 cells grown on different ECM substrates (fibronectin, laminin, and collagen) were analyzed. While N-cadherin, \u3b1SMA, and type I collagen were not significantly affected by ECM components, the E-cadherin/\u3b2-catenin complex was highly expressed in all the experimental conditions, and E-cadherin was upregulated by collagen in PL45 cells. Cell migration was unaffected by fibronectin and delayed by laminin. In contrast, collagen significantly stimulated cell migration and the secretion of MMPs. This study\u2019s results showed that ECM components impacted cell migration and invasive potential differently. Collagen exerted a more evident effect, providing new insights into the understanding of the intricate interplay between ECM molecules and cancer cells, in order to find novel therapeutic targets for PDAC treatment

Cross-code comparison of the edge codes SOLPS-ITER, SOLEDGE2D and UEDGE in modelling a low-power scenario in the DTT

As reactor-level nuclear fusion experiments are approaching, a solution to the power exhaust issue in future fusion reactors is still missing. The maximum steady-state heat load that can be exhausted by the present technology is around 10 MW m-2. Different promising strategies aiming at successfully managing the power exhaust in reactor-relevant conditions such that the limit is not exceeded are under investigation, and will be tested in the Divertor Tokamak Test (DTT) experiment. Meanwhile, the design of tokamaks beyond the DTT, e.g. EU-DEMO/ARC, is progressing at a high pace. A strategy to work around the present lack of reactor-relevant data consists of exploiting modelling to reduce the uncertainty in the extrapolation in the design phase. Different simulation tools, with their own capabilities and limitations, can be employed for this purpose. In this work, we compare SOLPS-ITER, SOLEDGE2D and UEDGE, three state-of-the-art edge codes heavily used in power exhaust studies, in modelling the same DTT low-power, pure-deuterium, narrow heat-flux-width scenario. This simplified, although still reactor-relevant, testbed eases the cross-comparison and the interpretation of the code predictions, to identify areas where results differ and develop understanding of the underlying causes. Under the conditions investigated, the codes show encouraging agreement in terms of key parameters at both targets, including peak parallel heat flux (1%-45%), ion temperature (2%-19%), and inner target plasma density (1%-23%) when run with similar input. However, strong disagreement is observed for the remaining quantities, from 30% at outer mid-plane up to a factor 4-5 at the targets. The results primarily reflect limitations of the codes: the SOLPS-ITER plasma mesh not reaching the first wall, SOLEDGE2D not including ion-neutral temperature equilibration, and UEDGE enforcing a common ion-neutral temperature. Potential improvements that could help enhance the accuracy of the code models for future applications are also discussed

Morphological and molecular characterization of human gingival tissue overlying multiple oral exostoses

Gingival and osseous augmentations are reported as hypertrophic or hyperplastic reactions to different factors including chronic traumatisms and surgeries such as free gingival graft (FGG) that induce an abnormal growth of both hard and soft tissues in genetically predisposed subjects. Since an imbalance in collagen turnover plays a key role in the development of gingival overgrowth leading to an accumulation of collagen in gingival connective tissue, in this study we described the histological and molecular features of three oral overgrowths obtained from a 34-year-old woman previously operated for FGG in order to evaluate a possible relationship between exostoses and overgrown tissue. Healthy and overgrown gingiva were analyzed by histological methods, and the expression of genes and proteins involved in collagen synthesis, maturation, and degradation was assessed in cultured fibroblasts obtained from gingival fragments at the molecular level. Our results show that general morphology and collagen content were similar in healthy and overgrown gingivae. However, fibroblasts obtained from the overgrown gingiva revealed an anabolic phenotype characterized by an increased collagen turnover and maturation. These findings indicate that an exostosis could act as a mechanical stimulus stretching the overlying connective tissue and triggering an anabolic phenotype of gingival fibroblasts and suggest to use minimally invasive surgical techniques to avoid traumatizing the periosteal tissues for the eradication of the exostosis with minimal relapses

Drp1 overexpression induces desmin disassembling and drives kinesin-1 activation promoting mitochondrial trafficking in skeletal muscle

Mitochondria change distribution across cells following a variety of pathophysiological stimuli. The mechanisms presiding over this redistribution are yet undefined. In a murine model overexpressing Drp1 specifically in skeletal muscle, we find marked mitochondria repositioning in muscle fibres and we demonstrate that Drp1 is involved in this process. Drp1 binds KLC1 and enhances microtubule-dependent transport of mitochondria. Drp1-KLC1 coupling triggers the displacement of KIF5B from kinesin-1 complex increasing its binding to microtubule tracks and mitochondrial transport. High levels of Drp1 exacerbate this mechanism leading to the repositioning of mitochondria closer to nuclei. The reduction of Drp1 levels decreases kinesin-1 activation and induces the partial recovery of mitochondrial distribution. Drp1 overexpression is also associated with higher cyclin-dependent kinase-1 (Cdk-1) activation that promotes the persistent phosphorylation of desmin at Ser-31 and its disassembling. Fission inhibition has a positive effect on desmin Ser-31 phosphorylation, regardless of Cdk-1 activation, suggesting that induction of both fission and Cdk-1 are required for desmin collapse. This altered desmin architecture impairs mechanotransduction and compromises mitochondrial network stability priming mitochondria transport through microtubule-dependent trafficking with a mechanism that involves the Drp1-dependent regulation of kinesin-1 complex

Acid Sphingomyelinase Downregulation Enhances Mitochondrial Fusion and Promotes Oxidative Metabolism in a Mouse Model of Melanoma

Melanoma is the most severe type of skin cancer. Its unique and heterogeneous metabolism, relying on both glycolysis and oxidative phosphorylation, allows it to adapt to disparate conditions. Mitochondrial function is strictly interconnected with mitochondrial dynamics and both are fundamental in tumour progression and metastasis. The malignant phenotype of melanoma is also regulated by the expression levels of the enzyme acid sphingomyelinase (A-SMase). By modulating at transcriptional level A-SMase in the melanoma cell line B16-F1 cells, we assessed the effect of enzyme downregulation on mitochondrial dynamics and function. Our results demonstrate that A-SMase influences mitochondrial morphology by affecting the expression of mitofusin 1 and OPA1. The enhanced expression of the two mitochondrial fusion proteins, observed when A-SMase is expressed at low levels, correlates with the increase of mitochondrial function via the stimulation of the genes PGC-1alpha and TFAM, two genes that preside over mitochondrial biogenesis. Thus, the reduction of A-SMase expression, observed in malignant melanomas, may determine their metastatic behaviour through the stimulation of mitochondrial fusion, activity and biogenesis, conferring a metabolic advantage to melanoma cells

Integration concept of an Electron Cyclotron System in DEMO

The pre-conceptual layout for an electron cyclotron system (ECS) in DEMO is described. The present DEMO ECS considers only equatorial ports for both plasma heating and neoclassical tearing mode (NTM) control. This differs from ITER, where four launchers in upper oblique ports are dedicated to NTM control and one equatorial EC port for heating and current drive (H&CD) purposes as basic configuration. Rather than upper oblique ports, DEMO has upper vertical ports to allow the vertical removal of the large breeding blanket segments. While ITER is using front steering antennas for NTM control, in DEMO the antennas are recessed behind the breeding blanket and called mid-steering antennas, referred to the radially recessed position to the breeding blanket. In the DEMO pre-conceptual design phase two variants are studied to integrate the ECS in equatorial ports. The first option integrates waveguide bundles at four vertical levels inside EC port plugs with antennas with fixed and movable mid-steering mirrors that are powered by gyrotrons, operating at minimum two different multiples of the fundamental resonance frequency of the microwave output window. Alternatively, the second option integrates fixed antenna launchers connected to frequency step-tunable gyrotrons. The first variant is described in this paper, introducing the design and functional requirements, presenting the equatorial port allocation, the port plug design including its maintenance concept, the basic port cell layout, the transmission line system with diamond windows from the tokamak up to the RF building and the gyrotron sources. The ECS design studies are supported by neutronic and tokamak integration studies, quasi-optical and plasma physics studies, which will be summarized. Physics and technological gaps will be discussed and an outlook to future work will be given

Essential role for acid sphingomyelinase-inhibited autophagy in melanoma response to cisplatin

The sphingolipid metabolising enzyme Acid Sphingomyelinase (A-SMase) has been recently shown to inhibit melanoma progression and correlate inversely to tumour grade. In this study we have investigated the role of A-SMase in the chemo-resistance to anticancer treatmentusing mice with melanoma allografts and melanoma cells differing in terms of expression/activity of A-SMase. Since autophagy is emerging as a key mechanism in tumour growth and chemo-resistance, we have also investigated whether an action of A-SMase in autophagy can explain its role. Melanoma sensitivity to chemotherapeutic agent cisplatin in terms of cell viability/apoptosis, tumour growth, and animal survival depended directly on the A-SMase levels in tumoural cells. A-SMase action was due to inhibition of autophagy through activation of Akt/mammalian target of rapamycin (mTOR) pathway. Treatment of melanoma-bearing mice with the autophagy inhibitor chloroquine restored sensitivity to cisplatin of tumours expressing low levels of A-SMase while no additive effects were observed in tumours characterised by sustained A-SMase levels. The fact that A-SMase in melanomas affects mTOR-regulated autophagy and plays a central role in cisplatin efficacy encourages pre-clinical testing on the modulation of A-SMase levels/activity as possible novel anti-neoplastic strategy

Integration Concept of an Electron Cyclotron System in DEMO

The pre-conceptual layout for an electron cyclotron system (ECS) in DEMO is described. The present DEMO ECS considers only equatorial ports for both plasma heating and neoclassical tearing mode (NTM) control. This differs from ITER, where four launchers in upper oblique ports are dedicated to NTM control and one equatorial EC port for heating and current drive (H&CD) purposes as basic configuration. Rather than upper oblique ports, DEMO has upper vertical ports to allow the vertical removal of the large breeding blanket segments. While ITER is using front steering antennas for NTM control, in DEMO the antennas are recessed behind the breeding blanket and called mid-steering antennas, referred to the radially recessed position to the breeding blanket.In the DEMO pre-conceptual design phase two variants are studied to integrate the ECS in equatorial ports. The first option integrates waveguide bundles at four vertical levels inside EC port plugs with antennas with fixed and movable mid-steering mirrors that are powered by gyrotrons, operating at minimum two different multiples of the fundamental resonance frequency of the microwave output window. Alternatively, the second option integrates fixed antenna launchers connected to frequency step-tunable gyrotrons. The first variant is described in this paper, introducing the design and functional requirements, presenting the equatorial port allocation, the port plug design including its maintenance concept, the basic port cell layout, the transmission line system with diamond windows from the tokamak up to the RF building and the gyrotron sources.The ECS design studies are supported by neutronic and tokamak integration studies, quasi-optical and plasma physics studies, which will be summarized. Physics and technological gaps will be discussed and an outlook to future work will be given