Il trattamento riabilitativo e il recupero precoce dopo intervento di “wedge resection” polmonare per secondarismi da tumore osseo: uno studio osservazionale

Background. Il recupero funzionale precoce dopo intervento di wedge resection polmonare non è studiato in maniera approfondita, in particolare, sono carenti i dati relativi al recupero postoperatorio precoce dei pazienti sottoposti a tele procedura chirurgica in seguito a metastasi polmonari per tumore osseo primitivo. Obiettivo. Lo scopo di questo studio osservazionale è quello di descrivere il percorso fisioterapico dopo intervento di wedge resection polmonare, evidenziando il possibile recupero funzionale nel breve termine e ricercandone i possibili fattori prognostici. Metodi. Per valutare il recupero funzionale si è utilizzato il 1 minute sit to stand test (1MSTS), che è stato somministrato in sesta giornata postoperatoria. Per descrivere l’andamento del recupero il test è stato somministrato in fase preoperatoria e in tutte le giornate postoperatorie. Sono state condotte analisi di univariata per valutare l’associazione delle variabili demografiche, anamnestiche e cliniche con l’outcome primario. Risultati. Sono stati reclutati 13 pazienti, per un totale di 17 osservazioni poiché alcuni pazienti hanno ripetuto l’operazione. Nella sesta giornata il 41,2% dei pazienti aveva eseguito lo stesso o un numero maggiore di verticalizzazioni del 1MSTS fatte nella fase preoperatoria. Nella sesta giornata postoperatoria il 100% dei pazienti era riuscito ad eseguire il 1MSTS almeno una volta dopo l’intervento. All’outcome primario risultavano associarsi con una significatività statistica il livello di autonomia, misurato con la TESS, (p=0,01) e il 1MSTS (p=0,001) misurati nella fase preoperatoria. Conclusioni. Il recupero funzionale postoperatorio è possibile fin dalle prime giornate postoperatorie per i pazienti sottoposti a wedge resection. Il livello di autonomia e il numero di verticalizzazioni del 1MSTS eseguite in fase preoperatoria sono possibili fattori prognostici del recupero funzionale postoperatorio

Revisiting the role of GSK3, a modulator of innate immunity, in idiopathic inclusion body myositis

Idiopathic or sporadic inclusion body myositis (IBM) is the leading age-related (onset > 50 years of age) autoimmune muscular pathology, resulting in significant debilitation in affected individuals. Once viewed as primarily a degenerative disorder, it is now evident that much like several other neuro-muscular degenerative disorders, IBM has a major autoinflammatory component resulting in chronic inflammation-induced muscle destruction. Thus, IBM is now considered primarily an inflammatory pathology. To date, there is no effective treatment for sporadic inclusion body myositis, and little is understood about the pathology at the molecular level, which would offer the best hopes of at least slowing down the degenerative process. Among the previously examined potential molecular players in IBM is glycogen synthase kinase (GSK)-3, whose role in promoting TAU phosphorylation and inclusion bodies in Alzheimer’s disease is well known. This review looks to re-examine the role of GSK3 in IBM, not strictly as a promoter of TAU and Abeta inclusions, but as a novel player in the innate immune system, discussing some of the recent roles discovered for this well-studied kinase in inflammatory-mediated pathology

Ankrd2/ARPP is a novel Akt2 specific substrate andregulates myogenic differentiation upon cellular exposure to H(2)O(2).

Activation of Akt-mediated signaling pathways is crucial for survival, differentiation, and regeneration of muscle cells. A proteomic-based search for novel substrates of Akt was therefore undertaken in C(2)C(12) murine muscle cells exploiting protein characterization databases in combination with an anti-phospho-Akt substrate antibody. A Scansite database search predicted Ankrd2 (Ankyrin repeat domain protein 2, also known as ARPP) as a novel substrate of Akt. In vitro and in vivo studies confirmed that Akt phosphorylates Ankrd2 at Ser-99. Moreover, by kinase assay with recombinant Akt1 and Akt2, as well as by single-isoform silencing, we demonstrated that Ankrd2 is a specific substrate of Akt2. Ankrd2 is typically found in skeletal muscle cells, where it mediates the transcriptional response to stress conditions. In an attempt to investigate the physiological implications of Ankrd2 phosphorylation by Akt2, we found that oxidative stress induced by H(2)O(2) triggers this phosphorylation. Moreover, the forced expression of a phosphorylation-defective mutant form of Ankrd2 in C(2)C(12) myoblasts promoted a faster differentiation program, implicating Akt-dependent phosphorylation at Ser-99 in the negative regulation of myogenesis in response to stress conditions

From interaction to function: Phospholipase C beta 1 protects cells from stress-induced apoptosis

The phosphoinositide-dependent signal transduction pathway has been implicated in the control of a variety of biological processes, such as the regulation of cellular metabolism and omeostasis, cell proliferation and differentiation. One of the key player in the regulation of inositol lipid signaling is phospholipase C beta 1 (PI-PLCβ1), which hydrolyses PtIns(4,5)P2, giving rise to the second messengers IP3 and DAG. The complete mapping of the PI-PLCβ1 interactome was undertaken, to understand its diverse functions within the nuclear compartment and to determine its contribution to physiological and pathological processes. Affinity purification-mass spectrometry (AP-MS) allowed for the identification of 160 proteins present in association with PI-PLCβ1 in the nucleus of erythroleukemia cells. Co-immunoprecipitation analysis of selected proteins confirmed the data obtained from mass spectrometry. Of particular interest was the identification of proteins involved in nuclear trafficking, as well as factors involved in hematological malignancies and several anti-apoptotic proteins (Piazzi et al., 2013). PI-PLCβ1 has been associated with the regulation of several cellular functions, some of which are not yet fully understood. In particular, it has been reported that PI-PLCβ1 protects murine fibroblasts from oxidative stress-induced cell death, through signaling events which remain unclear. Reactive oxygen species (ROS) have been shown to regulate major epigenetic processes causing the silencing of tumor suppressors and enhancing the proliferation of leukemic cells under oxidative stress. Investigation of the role for ROS and their signaling mediators in the pathogenesis of leukemia might, therefore, outline innovative approaches for the improvement of pharmacological therapies to treat leukemia. We demonstrate that in acute lymphoid leukemia cells (pro-B cells), treated with 250 μM of hydrogen peroxide (H2O2), PI-PLCβ1b conferred resistance to cell death, promoting cell cycle progression and cell proliferation. Interestingly, we found that, upon H2O2 exposure, the expression of PI-PLCβ1b affects the activity of several protein kinases, in particular it completely abolished the phosphorylation of Erk1/2 MAP kinases, down-regulated PTEN and up-regulated the phosphorylation of Akt; thereby sustaining cellular proliferation

How Inflammation Pathways Contribute to Cell Death in Neuro-Muscular Disorders

Neuro-muscular disorders include a variety of diseases induced by genetic mutations resulting in muscle weakness and waste, swallowing and breathing difficulties. However, muscle alterations and nerve depletions involve specific molecular and cellular mechanisms which lead to the loss of motor-nerve or skeletal-muscle function, often due to an excessive cell death. Morphological and molecular studies demonstrated that a high number of these disorders seem characterized by an upregulated apoptosis which significantly contributes to the pathology. Cell death involvement is the consequence of some cellular processes that occur during diseases, including mitochondrial dysfunction, protein aggregation, free radical generation, excitotoxicity and inflammation. The latter represents an important mediator of disease progression, which, in the central nervous system, is known as neuroinflammation, characterized by reactive microglia and astroglia, as well the infiltration of peripheral monocytes and lymphocytes. Some of the mechanisms underlying inflammation have been linked to reactive oxygen species accumulation, which trigger mitochondrial genomic and respiratory chain instability, autophagy impairment and finally neuron or muscle cell death. This review discusses the main inflammatory pathways contributing to cell death in neuro-muscular disorders by highlighting the main mechanisms, the knowledge of which appears essential in developing therapeutic strategies to prevent the consequent neuron loss and muscle wasting

Ectopic Expression of Ankrd2 Affects Proliferation, Motility and Clonogenic Potential of Human Osteosarcoma Cells

Simple Summary Osteosarcoma is a rare malignancy of bone, primarily affecting children and young adults. The main objective of this study was to identify novel therapeutic targets to fight the progression of this insidious disease. To this aim, the role of Ankrd2, a stress- and mechano- sensor protein known for being mostly expressed in muscle fibers, was analyzed in the modulation of osteosarcoma progression. By subjecting human osteosarcoma cell lines expressing or silencing Ankrd2 to several functional assays, our results demonstrated that Ankrd2 is involved in the pathogenesis of this cancer. Nonetheless, due to observations obtained by other studies in other model systems, our findings also suggest that Ankrd2 might behave as a "double-faced" cancer driver gene. Ankrd2 is a protein known for being mainly expressed in muscle fibers, where it participates in the mechanical stress response. Since both myocytes and osteoblasts are mesenchymal-derived cells, we were interested in examining the role of Ankrd2 in the progression of osteosarcoma which features a mechano-stress component. Although having been identified in many tumor-derived cell lines and -tissues, no study has yet described nor hypothesized any involvement for this protein in osteosarcoma tumorigenesis. In this paper, we report that Ankrd2 is expressed in cell lines obtained from human osteosarcoma and demonstrate a contribution by this protein in the pathogenesis of this insidious disease. Ankrd2 involvement in osteosarcoma development was evaluated in clones of Saos2, U2OS, HOS and MG63 cells stably expressing Ankrd2, through the investigation of hallmark processes of cancer cells. Interestingly, we found that exogenous expression of Ankrd2 influenced cellular growth, migration and clonogenicity in a cell line-dependent manner, whereas it was able to improve the formation of 3D spheroids in three out of four cellular models and enhanced matrix metalloproteinase (MMP) activity in all tested cell lines. Conversely, downregulation of Ankrd2 expression remarkably reduced proliferation and clonogenic potential of parental cells. As a whole, our data present Ankrd2 as a novel player in osteosarcoma development, opening up new therapeutic perspectives

Combined Treatment with PI3K Inhibitors BYL-719 and CAL-101 Is a Promising Antiproliferative Strategy in Human Rhabdomyosarcoma Cells

Rhabdomyosarcoma (RMS) is a highly malignant and metastatic pediatric cancer arising from skeletal muscle myogenic progenitors. Recent studies have shown an important role for AKT signaling in RMS progression. Aberrant activation of the PI3K/AKT axis is one of the most frequent events occurring in human cancers and serves to disconnect the control of cell growth, survival, and metabolism from exogenous growth stimuli. In the study reported here, a panel of five compounds targeting the catalytic subunits of the four class I PI3K isoforms (p110α, BYL-719 inhibitor; p110β, TGX-221 inhibitor; p110γ, CZC24832; p110δ, CAL-101 inhibitor) and the dual p110α/p110δ, AZD8835 inhibitor, were tested on the RMS cell lines RD, A204, and SJCRH30. Cytotoxicity, cell cycle, apoptosis, and the activation of downstream targets were analyzed. Of the individual inhibitors, BYL-719 demonstrated the most anti-tumorgenic properties. BYL-719 treatment resulted in G1/G0 phase cell cycle arrest and apoptosis. When combined with CAL-101, BYL-719 decreased cell viability and induced apoptosis in a synergistic manner, equaling or surpassing results achieved with AZD8835. In conclusion, our findings indicate that BYL-719, either alone or in combination with the p110δ inhibitor, CAL-101, could represent an efficient treatment for human rhabdomyosarcoma presenting with aberrant upregulation of the PI3K signaling pathway

The Cytotoxic Effect of Curcumin in Rhabdomyosarcoma Is Associated with the Modulation of AMPK, AKT/mTOR, STAT, and p53 Signaling

Approximately 7% of cancers arising in children and 1% of those arising in adults are soft tissue sarcomas (STS). Of these malignancies, rhabdomyosarcoma (RMS) is the most common. RMS survival rates using current therapeutic protocols have remained largely unchanged in the past decade. Thus, it is imperative that the main molecular drivers in RMS tumorigenesis are defined so that more precise, effective, and less toxic therapies can be designed. Curcumin, a common herbal supplement derived from plants of the Curcuma longa species, has an exceptionally low dietary biotoxicity profile and has demonstrated anti-tumorigenic benefits in vitro. In this study, the anti-tumorigenic activity of curcumin was assessed in rhabdomyosarcoma cell lines and used to identify the major pathways responsible for curcumin’s anti-tumorigenic effects. Curcumin treatment resulted in cell cycle arrest, inhibited cell migration and colony forming potential, and induced apoptotic cell death. Proteome profiler array analysis demonstrated that curcumin treatment primarily influenced flux through the AKT-mammalian target of rapamycin (mTOR), signal transducer and activator of transcription (STAT), AMP-dependent kinase (AMPK), and p53 associated pathways in a rhabdomyosarcoma subtype-specific manner. Thus, the strategic, combinational therapeutic targeting of these pathways may present the best option to treat this group of tumors

PLC-beta 1 regulates the expression of miR-210 during mithramycin-mediated erythroid differentiation in K562 cells

PLC-beta 1 (PLCβ1) inhibits erythroid differentiation induced by mithramycin (MTH) by targeting miR-210 expression. MicroRNA-210 (miR-210) has been reported to be upregulated in various types of human malignancy suggesting that it has an important role in tumorigenesis. Inhibition of miR-210 affects the erythroid differentiation pathway and it occurs to a greater extent in MTH-treated cells. In this paper we have analyzed the effect of MTH on human K562 cells differentiation. Overexpression of PLCβ1 suppresses the differentiation of K562 elicited by MTH as demonstrated by the absence of γ-globin expression. Inhibition of PLCβ1 expression is capable to promote the differentiation process leading to a recovery of γ-globin gene even in the absence of MTH. Our experimental evidences suggest that PLCβ1 signalling regulates erythropoiesis through miR-210. Indeed overexpression of PLCβ1 leads to a decrease of miR-210 expression after MTH treatment. Moreover miR-210 is up-regulated through both proliferation and differentiation events when PLCβ1 expression is down-regulated. Therefore we suggest a novel role for PLCβ1 in regulating miR-210 and our data hint at the fact that, in human K562 erythroleukemia cells, the modulation of PLCβ1 expression is able to exert an impairment of normal erythropoiesis as assessed by γ-globin expression