Search

This work aims at identifying the historical, cultural, political and strategic reasons that in 1935 led the anarchist Camillo Berneri to write the Constitution of the Italian Federation of Socialist Municipalities (based on the Italian acronym, FICS), an original constitution project with no predecessors within the anarchist movement which, unsurprisingly, remained unpublished for a long time. The research aims at explaining its existence within the Berneri’s intellectual path, interpreting it as the result of a long elaboration of his ideas and operational proposals on libertarian federalism. This maturation is characterized by the presence of different influences, with those shared with Gaetano Salvemini’s concrete and problematic approach being the decisive ones.Il lavoro si propone di individuare le motivazioni storiche, culturali, politiche e strategiche che hanno condotto l’anarchico Camillo Berneri a redigere nel 1935 la Costituzione della Federazione Italiana Comuni Socialisti (FICS), un progetto di costituzione originale senza precedenti all’interno del movimento anarchico e, non a caso, rimasto a lungo inedito. La ricerca intende rendere ragione della sua esistenza all’interno del percorso intellettuale berneriano, interpretandolo come esito di una lunga elaborazione delle idee e delle proposte operative di Berneri intorno al federalismo libertario. Tale maturazione risulta caratterizzata dalla presenza di diverse influenze, tra le quali sono determinanti quelle mutuate dall’approccio concretista e problemista di Gaetano Salvemini.

Background In order to identify suitable therapeutic targets for glioma anti-angiogenic therapy, the process of neovascularization mediated by circulating angiogenic cells (CACs) needs to be scrutinized. Methods In the present study, we compared the expression of neovascularization-related genes by 3 circulating CAC subsets (hematopoietic progenitor cells [HPCs], CD34+, and KDR+ cells; internal controls: peripheral blood mononuclear cells and circulating endothelial cells) of treatment-naïve patients with glioblastoma (GBM) to those of patients undergoing reactive neovascularization (myocardial infarction (MI). CACs from umbilical cord (representing developmental neovascularization) and healthy subjects served as controls. Fluorescent-activated cell sorting was used to isolate CACs, RT-PCR to determine the expression levels of a panel of 48 neovascularization-related genes, and Luminex assays to measure plasma levels of 21 CAC-related circulating molecules. Results We found essential differences in gene expression between GBM and MI CACs. GBM CACs had a higher expression of proangiogenic factors (especially, KITL, CXCL12, and JAG1), growth factor and chemotactic receptors (IGF1R, TGFBR2, CXCR4, and CCR2), adhesion receptor monomers (ITGA5 and ITGA6), and matricellular factor POSTN. In addition, we found major differences in the levels of neovascularization-related plasma factors. A strong positive correlation between plasma MMP9 levels and expression of CXCR4 in the CAC subset of HPCs was found in GBM patients. Conclusions Our findings indicate that CAC-mediated neovascularization in GBM is characterized by more efficient CAC homing to target tissue and a more potent proangiogenic response than in physiologic tissue repair in MI. Our findings can aid in selecting targets for therapeutic strategies acting against GBM-specific CACs.

Objective The gross majority of colorectal cancer cases results from aberrant Wnt/β-catenin signalling through adenomatous polyposis coli ( APC) or CTNNB1 mutations. However, a subset of human colon tumours harbour, mutually exclusive with APC and CTNNB1 mutations, gene fusions in RSPO2 or RSPO3, leading to enhanced expression of these R-spondin genes. This suggested that RSPO activation can substitute for the most common mutations as an alternative driver for intestinal cancer. Involvement of RSPO3 in tumour growth was recently shown in RSPO3 -fusion-positive xenograft models. The current study determines the extent into which solely a gain in RSPO3 actually functions as a driver of intestinal cancer in a direct, causal fashion, and addresses the in vivo activities of RSPO3 in parallel. Design We generated a conditional Rspo3 transgenic mouse model in which the Rspo3 transgene is expressed upon Cre activity. Cre is provided by cross-breeding with Lgr5 -GFP-Cre ERT2 mice. Results Upon in vivo Rspo3 expression, mice rapidly developed extensive hyperplastic, adenomatous and adenocarcinomatous lesions throughout the intestine. RSPO3 induced the expansion of Lgr5 + stem cells, Paneth cells, non-Paneth cell label-retaining cells and Lgr4 + cells, thus promoting both intestinal stem cell and niche compartments. Wnt/β-catenin signalling was modestly increased upon Rspo3 expression and mutant Kras synergised with Rspo3 in hyperplastic growth. Conclusions We provide in vivo evidence that RSPO3 stimulates the crypt stem cell and niche compartments and drives rapid intestinal tumorigenesis. This establishes RSPO3 as a potent driver of intestinal cancer and proposes RSPO3 as a candidate target for therapy in patients with colorectal cancer harbouring RSPO3 fusions.

Although extensive angiogenesis takes place in glial tumors, antiangiogenic therapies have remained without the expected success. In the peripheral circulation of glioma patients, increased numbers of endothelial precursor cells (EPCs) are present, potentially offering targets for antiangiogenic therapy. However, for an antiangiogenic therapy to be successful, the therapy should specifically target glioma-related EPC subsets and secreted factors only. Here, we compared the EPC subsets and plasma factors in the peripheral circulation of patients with gliomas to acute myocardial infarctions. We investigated the five most important EPC subsets and 21 angiogenesis-related plasma factors in peripheral blood samples of 29 patients with glioma, 14 patients with myocardial infarction, and 20 healthy people as controls, by FACS and Luminex assay. In GBM patients, all EPC subsets were elevated as compared to healthy subjects. In addition, HPC and KDR+ cell fractions were higher than in MI, while CD133+ and KDR+CD133+ cell fractions were lower. There were differences in relative EPC fractions between the groups: KDR+ cells were the largest fraction in GBM, while CD133+ cells were the largest fraction in MI. An increase in glioma malignancy grade coincided with an increase in the KDR+ fraction, while the CD133+ cell fraction decreased relatively. Most plasma angiogenic factors were higher in GBM than in MI patients. In both MI and GBM, the ratio of CD133+ HPCs correlated significantly with elevated levels of MMP9. In the GBM patients, MMP9 correlated strongly with levels of all HPCs. In conclusion, the data demonstrate that EPC traffic in patients with glioma, representing neoplasia, is different from that in myocardial infarction, representing tissue regeneration. Glioma patients may benefit from therapies aimed at lowering KDR+ cells and HPCs.

Human embryonal carcinoma (EC) cells comprise the pluripotent stem cells of malignant non-seminomatous germ cell tumors (GCTs) and represent the malignant counterpart of embryonic stem cells (ESCs). WNT/β-catenin signaling has been implicated in regulating adult and embryonic stem cells although its role in EC cells is less investigated. Here, we studied WNT signaling in a panel of representative pluripotent and nullipotent human EC cell lines. We found that EC cell lines show distinct levels of intrinsic WNT signaling and respond differently to ectopic WNT activation. Short-term activation of WNT signaling induced a differentiation-response in the pluripotent EC cells (NT2 and NCCIT) whereas the nullipotent EC cells (TERA1 and 2102Ep) were refractory and maintained high levels of OCT4 and SSEA4 expression. Long-term activation of WNT signaling in NCCIT and, to a lesser extent, TERA1 cells led to (re)gain of OCT4 expression and a switch from SSEA4 to SSEA1 surface antigens ultimately resulting in OCT4+/SSEA4−/SSEA1+ profile. Cisplatin treatment indicated that the OCT4+/SSEA4−/SSEA1+ NCCIT cells became more resistant to chemotherapy treatment. Our findings are of particular interest for the GCT and ES cell biology and shed light on the role of WNT signaling in human EC cells.

In this paper we consider a two-mode dynamical system as a model for a driven one-dimensional damped Bose–Einstein condensate in a double-well trapping potential. In the case of a constant external driving force the existence and stability of stationary solutions are discussed in relation to the values of the physical parameters. In the case of a time-dependent periodic external driving force the existence of limit cycles is proved, and the amplitude of these limit cycles exhibits a jump phenomenon for critical values of the physical parameters.

In this paper we give the \emph {quantization rules} to determine the normalized stationary solutions to the cubic nonlinear Schr\"odinger equation with quasi-periodic conditions on a given interval. \ Similarly to what happen in the Floquet's theory for linear periodic operators, also in this case some kind of band functions there exist., Comment: 14 pages - 6 figures

IBD syndromes such as Crohn's disease and ulcerative colitis result from the inflammation of specific intestinal segments. Although many studies have reported on the regenerative response of intestinal progenitor and stem cells to tissue injury, very little is known about the response of differentiated lineages to inflammatory cues. Here, we show that acute inflammation of the mouse small intestine is followed by a dramatic loss of Lgr5+ stem cells. Instead, Paneth cells re-enter the cell cycle, lose their secretory expression signature, and acquire stem-like properties, thus contributing to the tissue regenerative response to inflammation. Stem cell factor secretion upon inflammation triggers signaling through the c-Kit receptor and a cascade of downstream events culminating in GSK3β inhibition and Wnt activation in Paneth cells. Hence, the plasticity of the intestinal epithelium in response to inflammation goes well beyond stem and progenitor cells and extends to the fully differentiated and post-mitotic Paneth cells. Schmitt et al. show that inflammation of the mouse small intestine results in dramatic loss of the resident stem cells, followed by de-differentiation of highly specialized secretory units called Paneth cells. These findings are relevant for our understanding of tissue response in inflammatory bowel syndromes and their increased cancer risk.

The intestinal epithelium is characterized by an extremely rapid turnover rate. In mammals, the entire epithelial lining is renewed within 4 - 5 days. Adult intestinal stem cells reside at the bottom of the crypts of Lieberkühn, are earmarked by expression of the Lgr5 gene, and preserve homeostasis through their characteristic high proliferative rate1. Throughout the small intestine, Lgr5(+) stem cells are intermingled with specialized secretory cells called Paneth cells. Paneth cells secrete antibacterial compounds (i.e., lysozyme and cryptdins/defensins) and exert a controlling role on the intestinal flora. More recently, a novel function has been discovered for Paneth cells, namely their capacity to provide niche support to Lgr5(+ )stem cells through several key ligands as Wnt3, EGF, and Dll12. When isolated ex vivo and cultured in the presence of specific growth factors and extracellular matrix components, whole intestinal crypts give rise to long-lived and self-renewing 3D structures called organoids that highly resemble the crypt-villus epithelial architecture of the adult small intestine3. Organoid cultures, when established from whole crypts, allow the study of self-renewal and differentiation of the intestinal stem cell niche, though without addressing the contribution of its individual components, namely the Lgr5(+ )and Paneth cells. Here, we describe a novel approach to the organoid assay that takes advantage of the ability of Paneth and Lgr5(+) cells to associate and form organoids when co-cultured. This approach, here referred to as "organoid reconstitution assay" (ORA), allows the genetic and biochemical modification of Paneth or Lgr5(+) stem cells, followed by reconstitution into organoids. As such, it allows the functional analysis of the two main components of the intestinal stem cell niche.

Cancer stem–like cells (CSC) may be critical to maintain the malignant behavior of solid and hematopoietic cancers. Recently, patients with endometrial cancer whose tumors expressed high levels of aldehyde dehydrogenase (ALDH), a detoxifying enzyme characteristic of many progenitor and stem cells, exhibited a relative reduction in survival compared with patients with low levels of ALDH. Given evidence of its role as a CSC marker, we hypothesized that high level of ALDH activity (ALDHhi) in a tumor might positively correlate with the presence of stem- and progenitor-like tumor cells in this disease setting. In support of this hypothesis, ALDH could be used to enrich for CSC in endometrial cancer cell lines and primary tumors, as illustrated by the increased tumor-initiating capacity of ALDHhi cells in immunodeficient mice. ALDHhi cells also exhibited greater clonogenic and organoid-forming capacity compared with ALDHlo cells. Notably, the number of ALDHhi cells in tumor cell lines and primary tumors inversely correlated with differentiation grade. Expression analysis revealed upregulation of IL6 receptor subunits and signal transducers CD126 and GP130 in ALDHhi endometrial cancer cells. Accordingly, targeted inhibition of the IL6 receptor and its downstream effectors JAK1 and STAT3 dramatically reduced tumor cell growth. Overall, our results provide a preclinical rationale to target IL6 or its effector functions as a novel therapeutic option in endometrial cancer. Cancer Res; 75(17); 3608–22. ©2015 AACR.

Causal genetic changes in oligodendrogliomas (OD) with 1p/19q co-deletion include mutations in IDH1, IDH2, CIC, FUBP1, TERT promoter and NOTCH1. However, it is generally assumed that more somatic mutations are required for tumorigenesis. This study aimed to establish whether genes mutated at low frequency can be involved in OD initiation and/or progression. We performed whole-genome sequencing on three anaplastic ODs with 1p/19q co-deletion. To estimate mutation frequency, we performed targeted resequencing on an additional 39 ODs. Whole-genome sequencing identified a total of 55 coding mutations (range 8–32 mutations per tumor), including known abnormalities in IDH1, IDH2, CIC and FUBP1. We also identified mutations in genes, most of which were previously not implicated in ODs. Targeted resequencing on 39 additional ODs confirmed that these genes are mutated at low frequency. Most of the mutations identified were predicted to have a deleterious functional effect. Functional analysis on a subset of these genes (e.g. NTN4 and MAGEH1) showed that the mutation affects the subcellular localization of the protein (n = 2/12). In addition, HOG cells stably expressing mutant GDI1 or XPO7 showed altered cell proliferation compared to those expressing wildtype constructs. Similarly, HOG cells expressing mutant SASH3 or GDI1 showed altered migration. The significantly higher rate of predicted deleterious mutations, the changes in subcellular localization and the effects on proliferation and/or migration indicate that many of these genes functionally may contribute to gliomagenesis and/or progression. These low-frequency genes and their affected pathways may provide new treatment targets for this tumor type. Electronic supplementary material The online version of this article (doi:10.1007/s11060-015-1741-1) contains supplementary material, which is available to authorized users.

Background: Epidermal growth factor receptor (EGFR) is frequently mutated in various types of cancer. Although all oncogenic mutations are considered activating, different tumour types have different mutation spectra. It is possible that functional differences underlie this tumour-type specific mutation spectrum. Methods: We have determined whether specific mutations in EGFR (EGFR, EGFRvIII and EGFR-L858R) have differences in binding partners, differences in downstream pathway activation (gene expression and phosphoproteins), and have functional consequences on cellular growth and migration. Results: Using biotin pulldown and subsequent mass spectrometry we were able to detect mutation specific binding partners for EGFR. Differential binding was confirmed using a proximity ligation assay and/or Western Blot for the dedicator of cytokinesis 4 (DOCK4), UDP-glucose glycoprotein glucosyltransferase 1 (UGGT1), MYC binding protein 2 (MYCBP2) and Smoothelin (SMTN). We also demonstrate that each mutation induces the expression of a specific set of genes, and that each mutation is associated with specific phosphorylation patterns. Finally, we demonstrate using stably expressing cell lines that EGFRvIII and EGFL858R display reduced growth and migration compared to EGFR wildtype expressing cells. Conclusion: Our results indicate that there are distinct functional differences between different EGFR mutations. The functional differences between different mutations argue for the development of mutation specific targeted therapies. (C) 2015 Elsevier Ltd. All rights reserved.

Trop-2 is overexpressed in most human cancers, suggesting selective pressure for a key, conserved function. Here we show that Trop-2 stimulates cancer cell growth through the activation of a constitutively expressed, yet otherwise dormant, growth-control module. We discovered that crosslinking of membrane Trop-2 with specific Abs leads to a cytoplasmic Ca2+ raise through interaction with the Na+/K+-ATPase α1 subunit and mobilization of the intracellular stores. This triggers a feed-forward loop through Trop-2-dependent activation and membrane recruitment of PKCα;, which in turn phosphorylates the Trop-2 cytoplasmic tail at two target sites, activating the molecule to stimulate its downstream signaling targets Akt and ERK. Our findings indicate that the Trop-2-triggered cell growth operates through binding and extensive crosstalk with CD9, CD81, CD82 and CD151 through PKCα. Detailed analysis of CD9 and CD81 indicates that they bind to the HIKE region of the Trop-2 intracellular tail. Correspondingly, we found that the HIKE region of Trop-2 mediates its anchoring to the β-actin cytoskeleton through direct interaction with the ERM protein ezrin. Consistently, the Trop-2-dependent dynamic remodeling of the cell cytoskeleton is discovered to occur through activation of myosin II and binding of annexins A1/A11, α-actinin and gelsolin. Systematic drug screening, gene expression silencing and site-directed mutagenesis revealed that cytoskeleton disassembly, HIKE deletion and CD9 inhibition revert the growth of Trop-2-expressing cancer cells to that of their Trop-2-null counterparts. On the other hand, these inhibitors have no effects on basal cell growth. This indicates that Trop-2-centered protein interactions and activations are an essential step for the Trop-2-dependent cancer growth. Tight co-expression of the key components of the Trop-2 growth-stimulatory complex is found in a large breast cancer case series, thus indicating strong clinical relevance. Hence, Trop-2 triggers a universal, but otherwise dormant, layer of cancer growth, that overrides basal cell growth regulatory mechanisms and sensitizes tumors to targeted anticancer therapies. Citation Format: Marco Trerotola, Valeria Relli, Romina Tripaldi, Andrea Sacchetti, Kristina Havas, Pasquale Simeone, Emanuela Guerra, Annalaura Aloisi, Rossana La Sorda, Rossano Lattanzio, Daniele Vergara, Isabelle Fournier, Michel Salzet, Mauro Piantelli, Saverio Alberti. Trop-2 activates a dormant Na+/K+-ATPase/PKCα/CD9/ezrin signaling axis to override the basal growth program of cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 367. doi:10.1158/1538-7445.AM2017-367

In this paper we show that in the semiclassical regime of periodic potential large enough, the Stark-Wannier ladders become a dense energy spectrum because of a cascade of bifurcations while increasing the ratio between the effective nonlinearity strength and the tilt of the external field; this fact is associated to a transition from regular to quantum chaotic dynamics. The sequence of bifurcation points is explicitly given.

We discuss the method for the measurement of the gravity acceleration g by means of Bloch oscillations of an accelerated BEC in an optical lattice. This method has a theoretical critical point due to the fact that the period of the Bloch oscillations depends, in principle, on the initial shape of the BEC wavepacket. Here, by making use of the nearest-neighbor model for the numerical analysis of the BEC wavefunction, we show that in real experiments the period of the Bloch oscillations does not really depend on the shape of the initial wavepacket and that the relative uncertainty, due to the fact that the initial shape of the wavepacket may be asymmetrical, is smaller than the one due to experimental errors. Furthermore, we also show that the relation between the oscillation period and the scattering length of the BEC's atoms is linear; this fact suggests us a new experimental procedure for the measurement of the scattering length of atoms.

In this talk we consider the time evolution of a one-dimensional quantum system with a double barrier given by a couple of repulsive Dirac’s deltas. In such a pedagogical model we give, by means of the theory of quantum resonances, the asymptotic behavior of 〈ψ, e −itH ϕ〉 for large times, where H is the double-barrier Hamiltonian operator and where ψ and ϕ are two test functions. In particular, when ψ is close to a resonant state then explicit expression of the dominant terms of the survival probability defined as | 〈ψ, e −itH ψ〉 |2 is given.

The characterization of circulating endothelial progenitor cells (EPCs) is fundamental to any study related to angiogenesis. Unfortunately, current literature lacks consistency in the definition of EPC subsets due to variations in isolation strategies and inconsistencies in the use of lineage markers. Here we address critical points in the identification of hematopoietic progenitor cells (HPCs), circulating endothelial cells (CECs), and culture-generated outgrowth endothelial cells (OECs) from blood samples of healthy adults (AB) and umbilical cord (UCB). Peripheral blood mononuclear cells (PBMCs) were enriched using a Ficoll-based gradient followed by an optimized staining and gating strategy to enrich for the target cells. Sorted EPC populations were subjected to RT-PCR for tracing the expression of markers beyond the limits of cell surface-based immunophenotyping. Using CD34, CD133 and c-kit staining, combined with FSC and SSC, we succeeded in the accurate and reproducible identification of four HPC subgroups and found significant differences in the respective populations in AB vs. UCB. Co-expression analysis of endothelial markers on HPCs revealed a complex pattern characterized by various subpopulations. CECs were identified by using CD34, KDR, CD45, and additional endothelial markers, and were subdivided according to their apoptotic state and expression of c-kit. Comparison of UCB-CECs vs. AB-CECs revealed significant differences in CD34 and KDR levels. OECs were grown from PBMC-fractions We found that viable c-kit+ CECs are a candidate circulating precursor for CECs. RT-PCR to angiogenic factors and receptors revealed that all EPC subsets expressed angiogenesis-related molecules. Taken together, the improvements in immunophenotyping and gating strategies resulted in accurate identification and comparison of better defined cell populations in a single procedure.

BACKGROUND Although extensive angiogenesis takes place in glial tumors, anti-angiogenic therapies have remained without the expected success. In the peripheral circulation of glioma patients increased numbers of endothelial precursor cells (EPCs) are present, potentially offering targets for anti-angiogenic therapy (Zheng et al., Ann Neurol, 2007). However, for an anti-angiogenic therapy to be successful, the therapy should specifically target glioma-related EPC subsets and secreted factors. Here we compared the EPC subsets and plasma factors in the peripheral circulation of patients with gliomas to acute myocardial infarctions (representing fysiologic regeneration). MATERIAL AND METHODS We investigated the five most important EPC subsets and 21 angiogenesis-related plasma factors in peripheral blood samples of 29 patients with glioma, 14 patients with myocardial infarction and 20 healthy people as controls, by an advanced FACS protocol (Huizer et al., PlosOne 2018) and Luminex assay. RESULTS In GBM patients all EPC subsets were elevated as compared to healthy subjects. In addition, HPC and KDR+ cell fractions were higher than in MI, while CD133+ and KDR+CD133+ cell fractions were lower. There were differences in relative EPC fractions between the groups: KDR+ cells were the largest fraction in GBM while CD133+ cells were the largest fraction in MI. An increase in glioma malignancy grade coincided with an increase in the KDR+ fraction while the CD133+ cell fraction decreased relatively. Most plasma angiogenic factors were higher in GBM than MI patients. In both MI and GBM, the ratio of CD133+ HPCs correlated significantly with elevated levels of MMP9. In the GBM patients MMP9 correlated strongly with levels of all HPCs. CONCLUSION In conclusion, the data demonstrate that EPC traffic in patients with glioma is different from that in normal tissue regeneration. Therefore, the effects of glioma extent beyond the brain, and future therapies aimed at lowering KDR+ cells and HPCs may add to effective treatment.

Wnt signaling plays a central role in mammary stem cell (MaSC) homeostasis and in breast cancer. In particular, epigenetic alterations at different members of the Wnt pathway have been identified among triple-negative, basal-like breast cancers. Previously, we developed a mouse model for metaplastic breast adenocarcinoma, a subtype of triple-negative breast cancer, by targeting a hypomorphic mutations in the endogenous Apc gene (Apc (1572T/+)). Here, by employing the CD24 and CD29 cell surface antigens, we have identified a subpopulation of mammary cancer stem cells (MaCSCs) from Apc (1572T/+) capable of self-renewal and differentiation both in vivo and in vitro. Moreover, immunohistochemical analysis of micro- and macrolung metastases and preliminary intravenous transplantation assays suggest that the MaCSCs underlie metastasis at distant organ sites. Expression profiling of the normal and tumor cell subpopulations encompassing MaSCs and CSCs revealed that the normal stem cell compartment is more similar to tumor cells than to their own differentiated progenies. Accordingly, Wnt signaling appears to be active in both the normal and cancer stem cell compartments, although at different levels. By comparing normal with cancer mouse mammary compartments, we identified a MaCSC gene signature able to predict outcome in breast cancer in man. Overall, our data indicate that constitutive Wnt signaling activation affects self-renewal and differentiation of MaSCs leading to metaplasia and basal-like adenocarcinomas.

Among NSAIDs Celecoxib is one of the most efficient in triggering in vitro cancer cell death, and from this perspective has been subject of numerous studies. However, it is still controversial whether this in vitro-observed effect can also occur in vivo and contribute to the antitumor action of the drug. Moreover, besides common agreement on the involvement of COX-independent pathways, the mechanisms underlying Celecoxib toxicity are still unclear. In an attempt to shed light on these mechanisms, I found that cell death only occurs at insoluble concentrations of the drug, and follows irreversible binding and damage of the plasmamembrane by precipitates. This evidence strongly suggests that Celecoxib is devoid of true molecular toxicity. Moreover, since plasma levels reached during therapy are far below the threshold of toxic precipitation, direct cytotoxicity by Celecoxib is unlikely to occur on tumor cells in vivo. Thus the antitumor effect might be only due to COX inhibition, which requires significantly lower levels of the drug. Nonetheless, direct cytotoxicity might not be confined to an in vitro artifact, but contribute to the upper gastrointestinal side effects of Celecoxib. Overall, these findings represent an important basis for further studies on Celecoxib, where true molecular actions of the drug should be discriminated from the precipitate-dependent ones, and the relationship between in vitro and in vivo effects considered at the light of the precipitate-dependent model. Moreover, remarkably, this article indicates a model of critical analysis that can be extended to other poorly soluble drugs. J. Cell. Biochem. 114: 1434–1444, 2013. © 2013 Wiley Periodicals, Inc.

Purpose: Inhibition of AKT is a key target area for personalized cancer medicine. However, predictive markers of response to AKT inhibitors are lacking. Correspondingly, the AKT-dependent chain of command for tumor growth, which will mediate AKT-dependent therapeutic responses, remains unclear. Experimental Design: Proteomic profiling was utilized to identify nodal hubs of the Trop-2 cancer growth–driving network. Kinase-specific inhibitors were used to dissect Trop-2–dependent from Trop-2–independent pathways. In vitro assays, in vivo preclinical models, and case series of primary human breast cancers were utilized to define the mechanisms of Trop-2–driven growth and the mode of action of Trop-2–predicted AKT inhibitors. Results: Trop-2 and AKT expression was shown to be tightly coordinated in human breast cancers, with virtual overlap with AKT activation profiles at T308 and S473, consistent with functional interaction in vivo. AKT allosteric inhibitors were shown to only block the growth of Trop-2–expressing tumor cells, both in vitro and in preclinical models, being ineffective on Trop-2–null cells. Consistently, AKT-targeted siRNA only impacted on Trop-2–expressing cells. Lentiviral downregulation of endogenous Trop-2 abolished tumor response to AKT blockade, indicating Trop-2 as a mandatory activator of AKT. Conclusions: Our findings indicate that the expression of Trop-2 is a stringent predictor of tumor response to AKT inhibitors. They also support the identification of target-activatory pathways, as efficient predictors of response in precision cancer therapy. Clin Cancer Res; 22(16); 4197–205. ©2016 AACR.

A Bose-Einstein condensate (BEC) confined in a one-dimensional lattice under the effect of an external homogeneous field is described by the Gross-Pitaevskii equation. Here we prove that such an equation can be reduced, in the semiclassical limit and in the case of a lattice with a finite number of wells, to a finite-dimensional discrete nonlinear Schrodinger equation. Then, by means of numerical experiments we show that the BEC's center of mass exhibits an oscillating behavior with modulated amplitude; in particular, we show that the oscillating period actually depends on the shape of the initial wavefunction of the condensate as well as on the strength of the nonlinear term. This fact opens a question concerning the validity of a method proposed for the determination of the gravitational constant by means of the measurement of the oscillating period., 24 pages, 8 figures

We consider the stationary solutions for a class of Schroedinger equations with a symmetric double-well potential and a nonlinear perturbation. Here, in the semiclassical limit we prove that the reduction to a finite-mode approximation give the stationary solutions, up to an exponentially small term, and that symmetry-breaking bifurcation occurs at a given value for the strength of the nonlinear term. The kind of bifurcation picture only depends on the non-linearity power. We then discuss the stability/instability properties of each branch of the stationary solutions. Finally, we consider an explicit one-dimensional toy model where the double well potential is given by means of a couple of attractive Dirac's delta pointwise interactions., 46 pages, 4 figures

Accumulating evidence has suggested that tumours have a hierarchical organization in which only the cancer stem cells (CSCs) have tumour-initiating properties. Several surface antigens have been employed to isolate CSCs from various malignancies, although not from oesophageal adenocarcinoma (EA). We tested whether Barrett's oesophagus (BE) and EA might serve as a model for the CSC concept. In vivo assays were performed by transplantation of serially diluted bulk EA cells into NOD-SCID mice to establish the presence and frequency of tumour-initiating cells. These were found to be present as ca. 1 in 64 000 cells. The transplanted tumours fully recapitulated the primary lesions. Subsequently, a panel of previously established CSC markers was employed for immunohistochemistry. CD24, CD29 and CD44 showed heterogeneous staining in EA. Nuclear beta-catenin accumulation increased during progression from metaplasia to dysplasia and was often observed in the basal compartment with CD24 and CD29 staining. However, the overall staining patterns were not such to clearly point out specific candidate markers. Accordingly, all markers were employed to sort the corresponding subpopulations of cancer cells and transplant them at low multiplicities in NOD-SCID mice. No increased tumour-initiating capacity of sorted EA cells was observed upon transplantation. These results indicate that tumour-initiating cells are present in EA, thus reflecting a hierarchical organization. However, antibodies directed against novel surface antigens are needed to detect subpopulations enriched for CSCs in EA by transplantation assays.

We consider the Gross-Pitaevskii equation in 1 space dimension with a N-well trapping potential. We prove, in the semiclassical limit, that the finite dimensional eigenspace associated to the lowest N eigenvalues of the linear operator is slightly deformed by the nonlinear term into an almost invariant manifold $${\mathcal{M}}$$ . Precisely, one has that solutions starting on $${\mathcal{M}}$$ , or close to it, will remain close to $${\mathcal{M}}$$ for times exponentially long with the inverse of the size of the nonlinearity. As heuristically expected the effective equation on $${\mathcal M}$$ is a perturbation of a discrete nonlinear Schrodinger equation. We deduce that when the size of the nonlinearity is large enough then tunneling among the wells essentially disappears: that is for almost all solutions starting close to $${\mathcal{M}}$$ their restriction to each of the wells has norm approximatively constant over the considered time scale. In the particular case of a double well potential we give a more precise result showing persistence or destruction of the beating motions over exponentially long times. The proof is based on canonical perturbation theory; surprisingly enough, due to the Gauge invariance of the system, no non-resonance condition is required.

We consider the Gross-Pitaevskii equation in 1 space dimension with a N-well trapping potential. We prove, in the semiclassical limit, that the finite dimensional eigenspace associated to the lowest N eigenvalues of the linear operator is slightly deformed by the nonlinear term into an almost invariant manifold $${\mathcal{M}}$$ . Precisely, one has that solutions starting on $${\mathcal{M}}$$ , or close to it, will remain close to $${\mathcal{M}}$$ for times exponentially long with the inverse of the size of the nonlinearity. As heuristically expected the effective equation on $${\mathcal M}$$ is a perturbation of a discrete nonlinear Schrodinger equation. We deduce that when the size of the nonlinearity is large enough then tunneling among the wells essentially disappears: that is for almost all solutions starting close to $${\mathcal{M}}$$ their restriction to each of the wells has norm approximatively constant over the considered time scale. In the particular case of a double well potential we give a more precise result showing persistence or destruction of the beating motions over exponentially long times. The proof is based on canonical perturbation theory; surprisingly enough, due to the Gauge invariance of the system, no non-resonance condition is required.

Devices based on ultracold atoms moving in an accelerating optical lattice or double-well potential are a promising tool for precise measurements of fundamental physical constants as well as for the construction of sensors. Here, we carefully analyze the model of a couple of BECs separated by a barrier in an accelerated field and we show how the observable quantities, mainly the period of the beating motion or of the phase-shift, are related to the physical parameters of the model as well as to the energy of the initial state., 5 figures

In this paper we consider the time-independent one-dimensional non linear Schrodinger equation (NLS) with pointwise singular potential. We prove that when the strength of the pointwise interaction is less than a critical value, depending on the nonlinearity power σ , then a non linear real-valued bound state exists. Furthermore, we show that when σ is larger than 2 a further new real-valued stationary state appears under some conditions.

Current treatment options for glioblastoma patients have modest effects on survival. Since GBMs are one of the most vascularized tumors, treatment targeting GBM vessels has recently been introduced; clinical improvement, however, is limited. Endothelial progenitor cells are bone-marrow derived cells that home to ischemic tissue, guided by chemoattractants, where they differentiate under the influence of the microenvironment, and stimulate angiogenesis by secreting proangiogenic factors. As such, EPCs play an important role in neovascularization, including in GBMs. While changes in gene expression in EPCs once they arrive at their target location are well-established, little is known about the expression of proangiogenic factors in EPCs that are circulating in the blood. Since important factors from the GBM microenvironment also enter the bloodstream, we hypothesized that changes in gene expression could already be present in circulating EPCs from GBM patients. In order to test this hypothesis, we isolated EPCs (CD34 + CD133 + CD45dim) by FACS from blood of healthy controls (HC) and GBM patients. We determined the expression of a panel of 54 genes involved in angiogenesis by RT-PCR. The amount of circulating EPCs were similar between HC and GBM patients. Several genes, however, were differentially expressed; EPCs from GBM patients expressed both c-kit and SCF at a higher level than HC EPCs. In GBM, SCF is an important proangiogenic factor, which binds to its receptor c-kit. Both SCF and c-kit are expressed by subsets of endothelial cells in GBMs. We are currently investigating whether EPC c-kit and SCF expression correlates with tumor endothelial cell expression, and plan to do functional studies to determine the significance of our findings in GBM neovascularization. Our findings suggests that GBMs are capable of influencing gene expression in EPCs while they are still in the circulation. Influencing circulating EPCs in GBM patients could represent a novel type of antiangiogenic therapy.

In this paper we consider Bose-Einstein condensates (BECs) in one-, two- and three-dimension lattice potentials. The key argument for the explanation of the transition from Superfluidity phase to Mott-Insulator phase is suggested to be the spontaneous symmetry breaking effect which occurs for critical values of the ratio between the on-site interaction term and the hopping matrix element. Such an effect can be directly seen in the Gross-Pitaevskii equation with double-well potentials and it also explains the different behavior between one-dimensional models and two/three-dimensional models.

Here, we consider a two-level system driven by an external periodic field. We show that the coherent destruction of tunnelling, as proved by Grossmann and co-workers (1991 Phys. Rev. Lett. 67 516; 1992 Europhys. Lett. 18 571) in the case of a monochromatic field, also appears for any periodic driving field given by an even regular function with zero mean value and satisfying a technical condition on the zeros of this function.

Here, we show how the molecular structure appears and becomes stable for supercritical physical conditions. In particular we consider, for the ammonia molecule in a gas, a simple model based on a standard non-linear double-well Schroedinger equation with a dissipative term and a term representing weak collisions., Comment: 4 pages, 2 figures

Background.: The majority of glioma-associated microglia/macrophages have been identified as M2-type macrophages with immune suppressive and tumor supportive action. Recently, the extracellular adenosine deaminase protein Cat Eye Syndrome Critical Region Protein 1 (CECR1) was shown to regulate macrophage maturation. In this study, we investigate the role of CECR1 in the regulation of the glioma-associated macrophage response. Methods.: Expression of CECR1 was assessed in human glioma samples. CECR1-mediated macrophage response was studied in vitro, using donor derived CD14+ monocytes and the THP-1 monocytic cell line. The response of the human glioma cell line U87 to conditioned medium of macrophages preconditioned with recombinant human CECR1 or CECR1 silencing was also assessed. Results.: CECR1 was strongly expressed in high-grade gliomas (P < .001) and correlated positively with the M2 phenotype markers in tumor-associated microglia/macrophages (TAMs) (overall, P < .05). In vitro studies confirmed the presence of a significantly higher level of CECR1 expression in M2-like macrophages exposed to U87 conditioned medium (P < .001). CECR1 knockdown or stimulation of macrophages affected differentiation toward the M2-like phenotype. Stimulation of U87 cells with conditioned medium of CECR1 knockdown or stimulated macrophages affected tumor cell proliferation and migration, coinciding with altered intracellular signaling of mitogen-activated protein kinase (MAPK). In glioma tissue samples, CECR1 expression correlated with Ki67 and MAPK signaling protein. Conclusions.: CECR1 is a potent regulator of TAM polarization and is consistently highly expressed by M2-type TAMs, particularly in high-grade glioma. Paracrine effects induced by CECR1 in M2-like TAMs activate MAPK signaling and stimulate the proliferation and migration of glioma cells.

IntroductionAttempted suicide consists in a self-damaging non-fatal behaviour, with an explicit or implicit evidence of the intent to die. It has a multifactorial aetiology: presence of psychiatric disorder, particularly major depression and other non-psychiatric components, like a series of stressful events.AimTo describe suicidal behavior among patients who attempted suicide.MethodsCross-sectional study. Patients admitted to three hospitals in the Province of Modena (Italy) after having attempted suicide were enrolled. Observation time: August 2015–August 2016 (13 months). Descriptive statistics made with STATA 13.0.ResultsA total of 187 subjects (female 65%) were enrolled in the period of time considered. Forty-one percent were aged 45–64; 43.7% had higher education; 50.4% were unemployed; 49.6% had regular income. Common means to attempt suicide were drugs (64%), weapons (11%), precipitation (10%), other (6%), choking (4%). Recent stressful events were reported by the 83.4% of respondents, namely: family conflicts (29%), economic problems (17%), personal health problems (11%), health problems affecting a family member (5%), emotional separation (9%), job loss (7%), other (22%), alcohol abuse (27%), substance abuse (4%). The main psychiatric diagnoses were: depressive disorder (56.67%), personality disorder (20%), psychotic disorder (6.67%), bipolar disorder (5.33%), behavioural disorders (2.67%), anxiety disorders (2%), dementia (1.33%).ConclusionsFindings of the present study are consistent with available literature and could help to identify “high risk” groups to plan future targeted programmes.Disclosure of interestThe authors have not supplied their declaration of competing interest.

The small intestinal epithelium self-renews every four or five days. Intestinal stem cells (Lgr5(+) crypt base columnar cells (CBCs)) sustain this renewal and reside between terminally differentiated Paneth cells at the bottom of the intestinal crypt. Whereas the signalling requirements for maintaining stem cell function and crypt homeostasis have been well studied, little is known about how metabolism contributes to epithelial homeostasis. Here we show that freshly isolated Lgr5(+) CBCs and Paneth cells from the mouse small intestine display different metabolic programs. Compared to Paneth cells, Lgr5(+) CBCs display high mitochondrial activity. Inhibition of mitochondrial activity in Lgr5(+) CBCs or inhibition of glycolysis in Paneth cells strongly affects stem cell function, as indicated by impaired organoid formation. In addition, Paneth cells support stem cell function by providing lactate to sustain the enhanced mitochondrial oxidative phosphorylation in the Lgr5(+) CBCs. Mechanistically, we show that oxidative phosphorylation stimulates p38 MAPK activation by mitochondrial reactive oxygen species signalling, thereby establishing the mature crypt phenotype. Together, our results reveal a critical role for the metabolic identity of Lgr5(+) CBCs and Paneth cells in supporting optimal stem cell function, and we identify mitochondria and reactive oxygen species signalling as a driving force of cellular differentiation.

This paper presents the Microwave Imager (MWI) instrument, a microwave radiometer that is being developed for the MetOp Second Generation Programme. The MWI is a conically scanning radiometer and it will provide, in 2021-2042 timeframe, cloud and precipitation observations, snow and sea-ice coverage as well as gross profiles of water vapour and temperature.

In this paper we consider a one-dimensional non-linear Schrodinger equation with a periodic potential. In the semiclassical limit we prove the existence of stationary solutions by means of the reduction of the non-linear Schrodinger equation to a discrete non-linear Schrodinger equation. In particular, in the limit of large nonlinearity strength the stationary solutions turn out to be localized on a single lattice site of the periodic potential. A connection of these results with the Mott insulator phase for Bose–Einstein condensates in a one-dimensional periodic lattice is also discussed.

In this paper we prove the existence of the Stark-Wannier quantum resonances for one-dimensional Schrodinger operators with smooth periodic potential and small external homogeneous electric field. Such a result extends the existence result previously obtained in the case of periodic potentials with a finite number of open gaps., 30 pages, 1 figure

Here we consider the time evolution of a one-dimensional quantum system with a double barrier given by a couple of two repulsive Dirac's deltas. In such a "pedagogical" model we give, by means of the theory of quantum resonances, the explicit expression of the dominant terms of $$, where $H$ is the double-barrier Hamiltonian operator and where $\psi$ and $\phi$ are two test functions., Comment: 16 pages, 3 figures

In this paper we consider a two-level system under the effect of an external time-dependent field. We give a precise criterion for dynamical localization. We then apply our result to the cases of external ac-dc and bichromatic fields.

The Green Fluorescent Protein (GFP) is efficiently expressed in plant cells and its use as a flower fluorescent dye is appealing. However, the generation of fluorescent flowers has remained as yet elusive. In this work we demonstrate the generation of green-fluorescent flowers. Non-fluorescent GFP-transgenic flowers were analysed by Western blot and spectrofluorimetry, and were demonstrated to efficiently express GFP. This indicated that the lack of GFP fluorescence could be due to opacity to the exciting light. Thus, flowers that are largely transparent to UV, Eustoma grandiflorum (Lisianthus), or that present a transparent petal cuticle, Osteospermum ecklonis, were selected for gfp transformation. Strikingly, the transformed Eustoma flowers fluoresced brightly upon illumination with UV light. GFP-transgenic Osteospermum flowers also appeared distinctly green upon UV illumination. GFP was expressed in comparable amounts by the fluorescent Osteospermum and by the non-fluorescent Limonium petals, confirming that an efficient excitation of GFP fluorescence is a limiting factor in its detection. These results demonstrate the feasibility of using GFP as a fluorescent dye for flower petals. Chromophores excited at longer, more deeply penetrating wavelengths, e.g. DsRed, may extend the use this technology also to flowers opaque to UV light.

We study a periodically driven double well model. As in the case of autonomous models, previously treated in a joint paper with A. Martinez,(7) we have the destruction of the splitting for critical metastability. The relevance of the model for the understanding of the red shift in the inversion line of the molecule of ammonia is shortly discussed. We show that, in order to have a reasonable behavior of the metastability as a function of the frequency, a non-monochromatic perturbation is needed.

We consider the first-kind Fredholm integral equation , , where A is the Stieltjes transform defined as Under some regularity assumptions on f we prove that the above problem is well-posed according to Tikhonov; that is, for any f in a given class of data there exists a unique solution v of the above equation, and if , , for some positive then , , where is a continuous non-decreasing function with . An expression of the solution v by means of a convergent Fourier series is also given.

Magnesium homeostasis in HL-60 promyelocytic leukemia cells was compared to that in neutrophyl-like HL-60 cells obtained by 1.3% DMSO treatment. Magnesium homeostasis was studied by the characterization of magnesium efflux, the identification of intracellular magnesium pools, and the regulation of intracellular ionized Mg2+. In both undifferentiated and neutrophyl-like HL-60 cells, magnesium efflux occurred via the Na-Mg antiporter which was inhibited by imipramine and stimulated by db cAMP and forskolin. Receptor-mediated signals such as ATP, IFN-alpha, or PGE1, which can trigger cAMP-dependent magnesium efflux, were ineffective in undifferentiated HL-60 cells but induced 60-70% increase of magnesium efflux in neutrophyl-like HL-60 cells. Selective membrane permeabilization by the cation ionophore A23187 induced a large magnesium release when cells were treated with rotenone. In both cell populations, the addition of glucose to rotenone-treated cells restored magnesium release to the control level. Permeabilization by 0.005% digitonin provoked the release of 90% cell total magnesium in both cell types. Intracellular [Mg2+]i was 0.15 and 0.26 mM in undifferentiated and neutrophyl-like HL-60 cells, respectively. Stimuli that triggered magnesium efflux, such as db cAMP in undifferentiated and IFN-alpha in neutrophyl-like HL-60 cells, induced a slow but consistent increase of [Mg2+]i which was independent from Ca2+ movements. Overall, these data indicate that magnesium homeostasis is regulated by receptor-mediated magnesium efflux which was modified during differentiation of HL-60 cells. Stimulation of magnesium efflux is paralleled by an increase of [Mg2+]i which reflects a release of magnesium from the bound cation pool.