Type 1 Interferons Inhibit Myotube Formation Independently of Upregulation of Interferon-Stimulated Gene 15

Introduction: Type 1 interferon (IFN)-inducible genes and their inducible products are upregulated in dermatomyositis muscle. Of these, IFN-stimulated gene 15 (ISG15) is one of the most upregulated, suggesting its possible involvement in the pathogenesis of this disease. To test this postulate, we developed a model of type 1 IFN mediated myotube toxicity and assessed whether or not downregulation of ISG15 expression prevents this toxicity. Methods: Mouse myoblasts (C2C12 cell line) were cultured in the presence of type 1 or type 2 IFNs and ISG15 expression assessed by microarray analysis. The morphology of newly formed myotubes was assessed by measuring their length, diameter, and area on micrographs using imaging software. ISG15 expression was silenced through transfection with small interference RNA. Results: Type 1 IFNs, especially IFN-beta, increased ISG15 expression in C2C12 cells and impaired myotube formation. Silencing of ISG15 resulted in knockdown of ISG15 protein, but without phenotypic rescue of myotube formation. Discussion IFN-beta affects myoblast differentiation ability and myotube morphology in vitro.These studies provide evidence that ISG15, which is highly upregulated in dermatomyositis muscle, does not appear to play a key role in IFN-beta-mediated C2C12 myoblast cell fusion

Innate immunity in myasthenia gravis thymus: Pathogenic effects of Toll-like receptor 4 signaling on autoimmunity

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Neoadjuvant Chemo-Immunotherapy for Locally Advanced Non-Small-Cell Lung Cancer: A Review of the Literature

Non-small cell lung cancer accounts for approximately 80–85% of all lung cancers and at present represents the main cause of cancer death among both men and women. To date, surgery represents the cornerstone; nevertheless, around 40% of completely resected patients develop disease recurrence. Therefore, combining neoadjuvant chemo-immunotherapy and surgery might lead to improved survival. Immunotherapy is normally well tolerated, although significant adverse reactions have been reported in certain patients treated with inhibitors of immune checkpoints. In this review, we explore the current literature on the use of neoadjuvant chemo-immunotherapy followed by surgery for treatment of locally advanced non-small-cell lung cancer, with particular attention to the histological aspects, ongoing trials, and the most common surgical approaches. In conclusion, neoadjuvant immunotherapy whether combined or not with chemotherapy reveals a promising survival benefit for patients with advanced non-small-cell lung cancer; nevertheless, more data remain necessary to identify the best candidates for neoadjuvant regimens

Flood Directive Implemantation Methodology at Different Scales: Preliminary Results

In the frame of the application of 2007/60 flood directive in Italy, regional and basin authorities are presently carrying out activities concerning the implementation of risk maps, over the entire national territory. Basing on the different methodologies proposed in literature and by goverrnment agencies to calculate flood risk, a methodology is here proposed and applied in the Orco and Dora Riparia watershed basins. The methodology relies on available data, freeware software, free on-line documentation. It show to be applicable on different scales, from the watershed scale to that of urban district. Preliminary results are encouraging, as the methodology shows the robust, can be implemented in reasonable times and with less economical expenses. The risk index shows to be also relevant for decison making, especially when different optinos have to be compare

RNA silencing of ISG15 does not prevent IFN-β mediated myotoxicity.

<p>(A) Western blots of ISG15 demonstrate successful silencing of the 15 kDa ISG15 protein with siISG15 treatment in IFN-β treated C2C12 cells. Note absence of 15 kDa bands at 48 h (lane 2 compared to lane 1) and 72 h (lane 4 compared to lane 3), with partial return of protein expression at 96 h (lanes 6/5) and 120 h (lanes 8/7). siISG15 also reduces ISG15 conjugates (smears of >50 kDa) at all time points. Actin controls shown below. (B) Images demonstrate no improvement in myotube formation with ISG15 silencing at 72 h, and (C) quantitative analysis shows ISG15 silencing with siISG15 results in no recovery of myotube area at 72 h, 96 h, and 120 h.</p

Effects of type 1 IFNs on mouse C2C12 and human muscle cells.

<p>(A) IFN-β results in sustained marked expression of ISG15 (196-fold increased at Day 7). (B) Sustained toxicity of IFN-β on myotube area. (C–E) Dose-dependent effects of IFN-β 10 U/ml and 100 U/ml on myotubes. (C) Dose-dependent reduction in numbers and lengths of C2C12 myotubes at 48 h and 72 h. Arrows indicate myotubes. (D) Dose-dependent reduction in C2C12 myotube length, diameter, and area at 72h. (E) Dose-dependent effect of IFN-β on 72 h human skeletal muscle with marked inhibition of myotube formation at 100 U/ml.</p

CT after Lung Microwave Ablation: Normal Findings and Evolution Patterns of Treated Lesions

Imaging-guided percutaneous ablative treatments, such as radiofrequency ablation (RFA), cryoablation and microwave ablation (MWA), have been developed for the treatment of unresectable primary and secondary lung tumors in patients with advanced-stage disease or comorbidities contraindicating surgery. Among these therapies, MWA has recently shown promising results in the treatment of pulmonary neoplasms. The potential advantages of MWA over RFA include faster ablation times, higher intra-tumoral temperatures, larger ablation zones and lower susceptibility to the heat sink effect, resulting in greater efficacy in proximity to vascular structures. Despite encouraging results supporting its efficacy, there is a relative paucity of data in the literature regarding the role of computer tomography (CT) to monitor MWA-treated lesions, and the CT appearance of their morphologic evolution and complications. For both interventional and non-interventional radiologists, it is crucial to be familiar with the CT features of such treated lesions in order to detect incomplete therapy or recurrent disease at early stage, as well as to recognize initial signs of complications. The aim of this pictorial essay is to describe the typical CT features during follow-up of lung lesions treated with percutaneous MWA and how to interpret and differentiate them from other radiological findings, such as recurrence and complications, that are commonly encountered in this setting

Prediction of Ablation Volume in Percutaneous Lung Microwave Ablation: A Single Centre Retrospective Study

Background: Percutaneous Microwave Ablation (MWA) of lung malignancies is a procedure with many technical challenges, among them the risk of residual disease. Recently, dedicated software able to predict the volume of the ablated area was introduced. Cone-beam computed tomography (CBCT) is the imaging guidance of choice for pulmonary ablation in our institution. The volumetric prediction software (VPS) has been installed and used in combination with CBCT to check the correct position of the device. Our study aimed to compare the results of MWA of pulmonary tumours performed using CBCT with and without VPS. Methods: We retrospectively reviewed 1-month follow-up enhanced contrast-enhanced computed tomography (CECT) scans of 10 patients who underwent ablation with the assistance of VPS (group 1) and of 10 patients who were treated without the assistance of VPS (group 2). All patients were treated for curative purposes, the maximum axial diameter of lesions ranged between 5 and 22 mm in group 1 and between 5 and 25 mm in group 2. We compared the presence of residual disease between the two groups. Results: In group 1 residual disease was seen in only 1 patient (10%) in which VPS had ensured complete coverage of the tumour. In group 2 residual disease was found in 3 patients (30%). Conclusions: Using this software during MWA of lung malignancies could improve the efficacy of the treatment compared to the conventional only CBCT guidance

B Cell Lymphoma (Bcl)-2 Protein Is the Major Determinant in bcl-2 Adenine-Uridine-rich Element Turnover Overcoming HuR Activity*

In the 3′-untranslated region, the destabilizing adenine-uridine (AU)-rich elements (AREs) control the expression of several transcripts through interactions with ARE-binding proteins (AUBPs) and RNA degradation machinery. Although the fundamental role for AUBPs and associated factors in eliciting ARE-dependent degradation of cognate mRNAs has been recently highlighted, the molecular mechanisms underlying the specific regulation of individual mRNA turnover have not yet been fully elucidated. Here we focused on the post-transcriptional regulation of bcl-2 mRNA in human cell lines under different conditions and genetic backgrounds. In the context of an AUBPs silencing approach, HuR knockdown reduced the expression of endogenous bcl-2, whereas unexpectedly, a bcl-2 ARE-reporter transcript increased significantly, suggesting that HuR expression has opposite effects on endogenous and ectopic bcl-2 ARE. Moreover, evidence was provided for the essential, specific and dose-dependent role of the Bcl-2 protein in regulating the decay kinetics of its own mRNA, as ascertained by a luciferase reporter system. Altogether, the data support a model whereby the Bcl-2 protein is the major determinant of its own ARE-dependent transcript half-life in living cells and its effect overcomes the activity of ARE-binding proteins