Calpain restrains the stem cells compartment in breast cancer

CAPNS1 is essential for the stability and function of ubiquitous CAPN1 and CAPN2. Calpain modulates by proteolytic cleavage many cellular substrates and its activity is often deregulated in cancer cells, therefore calpain inhibition has been proposed as a therapeutical strategy for a number of malignancies. Here we show that CAPNS1 depletion is coupled to impairment of MCF7 and MCF10AT cell lines growth on plate and defective architecture of mammary acini derived from MCF10A cells. In soft agar CAPNS1 depletion leads to cell growth increase in MCF7, and decrease in MCF10AT cells. In both MCF7 and MCF10AT, CAPNS1 depletion leads to the enlargement of the stem cell compartment, as demonstrated by mammosphere formation assays and evaluation of stem cell markers by means of FACS and western blot analysis. Accordingly, activation of calpain by thapsigargin treatment leads to a decrease in the stem cell reservoir. The expansion of the cancer stem cell population in CAPNS1 depleted cells is coupled to a defective shift from symmetric to asymmetric division during mammosphere growth coupled to a decrease in NUMB protein level

Regulation of Sport Activities and Right to Respect to Private Life under the European Convention on Human Rights

In F\ue9d\ue9ration Nationale des Syndicats Sportifs (FNASS) and others v. France (judgment of 18 January 2018, no. 48151/11 and 77769/13), the European Court of Human Rights assessed the compatibility between the right to private and family life and a French Order regulating unannounced anti-doping controls for sport professionals. According to the European Court of Human Rights, the impact of the disputed regulation on sport professionals\u2019 right to private life is not disproportionate with respect to the legitimate necessity to protect public health and assure fairness and equality of opportunities among sport competitors. This decision confirms a specific approach characterising the, albeit limited, case-law elaborated by the European Court of Human Rights on the interplay between sport activities, on one hand, and the right to respect for private and family life, on the other. The European Court of Human Rights tends to highlight the leisure nature of sport activities more than the interests on the part of sport professionals. In relation to professional sport activities, the balancing test seems to place emphasis on public interest linked with the regulation of sport competitions more than on the right to privacy of sport professionals. Instead, when sport activities come into play in relation to their leisure nature, the European Court of Human Rights deeply stresses their importance as a means allowing persons to develop social relations and express personal identity

Il “battesimo del fuoco” della direttiva sui prodotti del tabacco, tra esigenze di armonizzazione e di tutela della salute pubblica

I. Introduzione. II. Esigenze di armonizzazione e obiettivi di politica sanitaria. III. Il bilanciamento tra la tutela della salute e altri diritti e libertà enunciati dalla Carta dei diritti fondamentali.On 4 May 2016, the ECJ delivered three judgments concerning the validity of the “Tobacco Products Directive”. This article focuses on the legal basis of this controversial act and on the fundamental rights’ restrictions provided for therein. As for the legal basis, the ECJ's judgments found that, in order to avoid the negative impact of divergent national legislations on the internal market, the EU is allowed to exercise its harmonizing competence in the field of public health, despite this policy being essentially reserved to the Member States. This finding was essentially based on ECJ’s previous case law. With regard to the issue of the fundamental rights’ restrictions raised by private parties in two of the judgments, the Court found these restrictions to be justified under the general limitation clause of art. 52 of the Charter. In more specific terms, the Court affirmed that, in the cases at hand, the EU was allowed to strike a fair balance between the need to respect fundamental rights and freedoms and the need to protect public health. This conclusion was predicated on the provisions of the TFEU and the Charter concerning the attainment of “a high level of protection” of human health. As suggested in this study, this approach of the ECJ can be read as part of a recent attitude towards referring more extensively to art. 35 of the Charter (“Health care”)

L\u2019integrazione europea, le giovani generazioni e l\u2019idea di una politica comune per la famiglia

In un'epoca in cui si avverte insoddisfazione e disinteresse verso gli ideali dell'integrazione europea, spesso anche tra i giovani, non va dimenticato che l'UE ha creato uno spazio in cui circolare, lavorare, studiare liberamente, che non ha precedenti nella storia d'Europa, che non ha eguali negli altri continenti e che si \ue8 allargato sempre pi\uf9 nel corso dei decenni. Ai cittadini dell\u2019UE, e specialmente alle giovani generazioni, occorre ricordare che l'UE ha determinato la creazione di una dimensione di libert\ue0 negli ultimi 60 anni che troppo spesso \ue8 data per scontata e alla quale non saprebbe rinunciare chi ha la fortuna di vivere sul territorio dell'Unione. Si possono per\uf2 certamente auspicare ulteriori progressi nella democraticit\ue0 delle Istituzioni europee, in politiche sociali pi\uf9 incisive, e soprattutto nell\u2019adozione di scelte politiche chiare di sostegno diretto alle famiglie.In times of dissatisfaction and disinterest in the ideals of European integration, even among young people, we should not forget that the EU has created an area of freedom where it is possible to circulate, work and study freely. This fact has no precedent in the history of Europe, no equal in any other continent and has expanded ever more over the decades. EU citizens, especially the younger ones, should be reminded that the EU has determined the creation of a dimension of freedom in the last 60 years which is too often taken for granted. To bring people closer to the EU, it would be better to further progress in the democracy of the EU institutions, taking more effective EU social measures, adopting clear EU political choices of direct support to families

Construction of miniantibodies for the in vivo study of human autoimmune diseases in animal models

<p>Abstract</p> <p>Background</p> <p>Phage display antibody libraries have been made from the lymphocytes of patients suffering from autoimmune diseases in which the antibodies are known to play a role in the pathogenesis or are important for the diagnosis of the disease. In the case of Celiac Disease, the immune response is directed against the autoantigen tissue transglutaminase. However, despite numerous studies, the role of these antibodies in the pathogenesis of this disease has not been elucidated.</p> <p>Results</p> <p>We were able to engineer specific anti-transglutaminase antibody fragments in the form called "miniantibody". These are produced by genetic fusion of anti-tTG scFv to Human, Mouse or Rat Fc domains, making them suitable for in vivo expression. The results obtained here indicate that the miniantibody molecule is efficiently secreted, and that the reactivity to the antigen is retained even after fusion to heterologous Fc domains. Further analysis demonstrate that the molecule is secreted as homodimeric, mimicking original antibody structure. Finally, the in vivo expression in mice leads to detectable serum levels with no apparent gross immune response by the host.</p> <p>Conclusion</p> <p>In this work we demonstrated the usefulness of a method for the in vivo expression of miniantibodies specific to transglutaminase, corresponding to the autoimmune specificity of Celiac Disease. This can be proposed as a general method to study the pathogenic role of autoimmune antibodies in autoimmune diseases.</p

Targeting of surface alpha-enolase inhibits the invasiveness of pancreatic cancer cells

Pancreatic Ductal Adenocarcinoma (PDAC) is a highly aggressive malignancy characterized by rapid progression, invasiveness and resistance to treatment. We have previously demonstrated that most PDAC patients have circulating antibodies against the glycolytic enzyme alpha-enolase (ENO1), which correlates with a better response to therapy and survival. ENO1 is a metabolic enzyme, also expressed on the cell surface where it acts as a plasminogen receptor. ENO1 play a crucial role in cell invasion and metastasis by promoting plasminogen activation into plasmin, a serine-protease involved in extracellular matrix degradation. The aim of this study was to investigate the role of ENO1 in PDAC cell invasion. We observed that ENO1 was expressed on the cell surface of most PDAC cell lines. Mouse anti-human ENO1 monoclonal antibodies inhibited plasminogen-dependent invasion of human PDAC cells, and their metastatic spreading in immunosuppressed mice was inhibited. Notably, a single administration of Adeno-Associated Virus (AAV)-expressing cDNA coding for 72/1 anti-ENO1 mAb reduced the number of lung metastases in immunosuppressed mice injected with PDAC cells. Overall, these data indicate that ENO1 is involved in PDAC cell invasion, and that administration of an anti-ENO1 mAb can be exploited as a novel therapeutic option to increase the survival of metastatic PDAC patients

Autologous anti-GD2 CAR T cells efficiently target primary human glioblastoma

Glioblastoma (GBM) remains a deadly tumor. Treatment with chemo-radiotherapy and corticosteroids is known to impair the functionality of lymphocytes, potentially compromising the development of autologous CAR T cell therapies. We here generated pre-clinical investigations of autologous anti-GD2 CAR T cells tested against 2D and 3D models of GBM primary cells. We detected a robust antitumor effect, highlighting the feasibility of developing an autologous anti-GD2 CAR T cell-based therapy for GBM patients

Assessment of the flotability of chalcopyrite, molybdenite and pyrite using biosolids and their main components as collectors for greening the froth flotation of copper sulphide ores.

Biosolids and representative compounds of their main components ? humic acids, sugars, and proteins ? have been tested as possible environment-friendly collectors and frothers for the flotation of copper sulphide ores. The floatability of chalcopyrite and molybdenite ? both valuable sulphide minerals present in these ores ? as well as non-valuable pyrite was assessed through Hallimond tube flotation tests. Humic acids exhibit similar collector ability for chalcopyrite and molybdenite as that of a commercial collector (Aero 6697 promoter). Biosolids show more affinity for pyrite. The copper recovery (85.9%) and copper grade (6.7%) of a rougher concentrate obtained using humic acids as main collector for the flotation of a copper sulphide ore from Chile, were very similar to those of a copper concentrate produced by froth flotation under the same conditions with a xanthate type commercial collector. This new and feasible end-use of biosolids and humic acids should be new environment-friendly organic froth flotation agents for greening the concentration of copper sulphide ore. Now, further research is needed in order to scale current laboratory assays to operational mining scales to determine efficiencies to industrial scale