Chloroquine supplementation increases the cytotoxic effect of curcumin against Her2/neu overexpressing breast cancer cells in vitro and in vivo in nude mice while counteracts it in immune competent mice

Autophagy is usually a pro-survival mechanism in cancer cells, especially in the course of chemotherapy, thus autophagy inhibition may enhance the chemotherapy-mediated anti-cancer effect. However, since autophagy is strongly involved in the immunogenicity of cell death by promoting ATP release, its inhibition may reduce the immune response against tumors, negatively influencing the overall outcome of chemotherapy. In this study, we evaluated the in vitro and in vivo anti-cancer effect of curcumin (CUR) against Her2/neu overexpressing breast cancer cells (TUBO) in the presence or in the absence of the autophagy inhibitor chloroquine (CQ). We found that TUBO cell death induced by CUR was increased in vitro by CQ and slightly in vivo in nude mice. Conversely, CQ counteracted the Cur cytotoxic effect in immune competent mice, as demonstrated by the lack of in vivo tumor regression and the reduction of overall mice survival as compared with CUR-treated mice. Immunohistochemistry analysis revealed the presence of a remarkable FoxP3 T cell infiltrate within the tumors in CUR/CQ treated mice and a reduction of T cytotoxic cells, as compared with single CUR treatment. These findings suggest that autophagy is important to elicit anti-tumor immune response and that autophagy inhibition by CQ reduces such response also by recruiting T regulatory (Treg) cells in the tumor microenvironment that may be pro-tumorigenic and might counteract CUR-mediated anti-cancer effects

The activation of KSHV lytic cycle blocks autophagy in PEL cells

This study confirms that autophagy is activated concomitantly with KSHV lytic cycle induction, and that autophagy inhibition by BECN1 knockdown reduces viral lytic gene expression. In addition, we extend previous observations and show that autophagy is blocked at late steps, during viral replication. This is indicated by the lack of colocalization of autophagosomes and lysosomes and by the LC3-II level that does not increase in the presence of bafilomycin A1 in primary effusion lymphoma (PEL) cells induced to enter the lytic cycle, either by TPA/sodium butyrate (BC3 and BCBL1) or by doxycycline (TRExBCBL1-Rta). The autophagic block correlates with the downregulation of RAB7, whose silencing with specific siRNA results in an autophagic block in the same cells. Finally, by electron microscopy analysis, we observed viral particles inside autophagic vesicles in the cytoplasm of PEL cells undergoing viral replication, suggesting that they may be involved in viral transpor

Universal hydrophilic coating of thermoplastic polymers currently used in microfluidics

A number of materials used to fabricate disposable microfluidic devices are hydrophobic in nature with water contact angles on their surface ranging from 80 to over 100. This characteristic makes them unsuitable for a number of microfluidic applications. Both the wettability and analyte adsorption parameters are highly dependent on the surface hydrophobicity. In this article, we propose a general method to coat the surface of five materials: polydimethylsiloxane (PDMS), cyclic olefin copolymer (COC), polyethylene terephthalate (PET), polycarbonate (PC), and polytetrafluoroethylene (PTFE). This fast and robust process, which is easily implementable in any laboratory including microfabrication clean room facilities, was devised by combining gas-phase and wet chemical modification processes. Two different coatings that improve the surface hydrophilicity were prepared via the "dip and rinse" approach by immersing the plasma oxidized materials into an aqueous solution of two different poly(dimethylacrylamide) copolymers incorporating a silane moiety and functionalized with either N-acryloyloxysuccinimide (NAS) (poly(DMA-NAS-MAPS) or glycidyl methacrylate (GMA) (poly(DMA-GMA-MAPS). The coating formation was confirmed by contact angle (CA) analysis comparing the variation of CAs of uncoated and coated surfaces subjected to different aging treatments. The antifouling character of the polymer was demonstrated by fluorescence and interferometric detection of proteins adsorbed on the surafce. This method is of great interest in microfluidics due to its broad applicability to a number of materials with varying chemical compositions

CAIMAN Experiment

Non-acoustic detection systems can be used in combination with Sonar systems to determine the presence of an underwater threat, such as terrorist divers. The goal of the CAIMAN (Coastal Anti Intruders MAgnetometers Network) joint experiment (Italian Navy, NATO Undersea Research Centre and INGV Marine Geophysics) is the application of High Definition Geophysics Magnetic techniques in a port protection scenario, where conventional measurements of very low magnetic sources, like intruder swimmers, are strongly disturbed by ambient, natural and artificial, background noise and other time-variant magnetic anomalies. Two tri-axial fluxgate magnetometers were deployed on the sea bottom and connected to a shore side measurement station. A team of navy divers, wearing both COTS and EOD equipment, performed some coastal approach runs on each magnetometer alternatively. Magnetic signature data were logged and post processed using MATLAB®. Results demonstrated the effectiveness of high definition time reduction techniques using a self-referred integrated array design

Misure ambientali in mare aperto: sviluppo di tecnologie per l'acquisizione e l'analisi di dati meteo-mareografici misurati da una boa oceanografica in Mar Ligure

Obiettivo del presente lavoro è lo sviluppo di un metodo di analisi in grado di stimare i parametri fondamentali del moto ondoso, a partire dai dati acquisiti da tre altimetri acustici installati a bordo di una boa meteo-oceanografica operante in mare aperto. Il metodo di analisi sviluppato compie opportune operazioni di filtraggio sulle sequenze temporali delle misure effettuate dai tre altimetri, quindi, elaborando tali sequenze, fornisce le stime di alcuni parametri caratteristici del moto ondoso (tra cui l’altezza e la direzione di propagazione). I diversi tipi di filtraggio ed il metodo di stima sviluppati si basano sul calcolo di alcuni parametri statistici (tra cui media, mediana e deviazione standard) delle serie temporali di dati acquisiti, sulla conoscenza delle loro densità spettrali di potenza (calcolate mediante FFT), e sul calcolo delle funzioni di crosscorrelazione delle sequenze di dati prese a due a due. Il procedimento di stima realizzato è stato sperimentato su una notevole quantità di dati reali acquisiti in Mar Ligure tramite l’utilizzo della stazione di misura fissa su cui sono montati gli altimetri acustici, ed ha fornito risultati soddisfacenti per quanto riguarda affidabilità e precisione. Nell’ambito della valutazione delle prestazioni del sistema di acquisizione dati e del metodo di stima, i risultati ottenuti sono stati confrontati con misure provenienti da altri sensori a bordo della stazione e con stime analoghe effettuate a partire dai dati acquisiti da un’altra stazione di misura, operante anch’essa in Mar Ligure, ma dotata di strumentazione di altro tipo

A Preliminary Study on SVM based Analysis of Underwater Magnetic Signals for Port Protection

People who attend to the problem of underwater port protection usually use sonar based systems. Recently it has been shown that integrating a sonar system with an auxiliary array of magnetic sensors can improve the effectiveness of the intruder detection system. One of the major issues that arise from the integrated magnetic and acoustic system is the interpretation of the magnetic signals coming from the sensors. In this paper a machine learning approach is proposed for the detection of divers or, in general, of underwater magnetic sources. The research proposed here, by means of a windowing of the signals, uses Support Vector Machines for classification, as tool for the detection problem. Empirical results show the effectiveness of the method

Marine geomagnetic high definition metrology; possible archaeological applications

The marine geomagnetism is often used to solve geological problems at a regional scale: structural, volcanological or mining problems, for instance (Faggioni et al., 1995). In recent time, the development of the “high definition (HD) metrology” (Faggioni et al., 2001) has made the marine geomagnetic method suitable for detecting short wavelength and low amplitude geomagnetic anomalies. This type of signal may be often related to environmental and/or archaeological sources......

Anti-intruder port protection “MAC (Magnetic ACoustic) System”: advances in the magnetic component performance

In the following, we report the advances made in the development of a magnetic component for peripheral monitoring of port environments for anti-intruder systems. The project was initiated by the Office of Research and Development, COMFORDRAG, Italian Navy, in 2004, with the aim of improving the detection performance of current acoustic-based anti-intruder systems, in the vicinity of quays or the seabed or in acoustic-shadow zones behind natural or artificial objects. In the five-year period from 2004 to 2008, the system has been subjected to detection-performance tests in experiments involving divers equipped with air tanks and rebreather kits in real port environments and, at the same time, the associated signal processing has been subjected to development, to enhance detection performance. In tests involving simulated diver attacks against a vessel moored alongside a quay in the port of La Spezia, the system has been proved particularly effective by detecting 9 divers out of 9. The processing of the magnetic signal has been enhanced by implementing an amplitude threshold, as a means of filtering the passive energy components, termed Passive Energy Cutting (PEC), that is, the elimination of components that comprise no useful information regarding diver transits. The use of PEC boosts the reliability of the system in terms of detection versus false alarm rate, as evidenced by the comparison of the ROC curves for the system with and without PEC

Carrageenan-induced acute inflammation in the mouse air pouch synovial model. Role of tumour necrosis factor

We used the mouse air pouch model of inflammation to study the interaction between cytokines, prostaglandin E2 (PGE2) and cell migration during the various phases of acute local inflammation induced by carrageenan. In serum, the levels of interleukin 1 (IL-1), interleukin 6 (IL-6), tumour necrosis factor (TNF), serum amiloid-A (SAA) and Fe++ were never different from controls, indicating that no systemic inflammatory changes were induced. Locally the exudate volume and the number of leukocytes recruited into the pouch increased progressively until 7 days after carrageenan. The same was true for PGE2 production. We could not measure IL-1 but the production of IL-6 and TNF reached a maximum after 5-24 h then quickly decreased. Anti-TNF antibodies inhibited cell migration by 50% 24 h after treatment. Pretreatment with interleukin 10 (IL-10) inhibited TNF production almost completely and cell migration by 60%. Carrageenan-induced inflammation was modulated by anti-inflammatory drugs. Pretreatment with dexamethasone (DEX) or indomethacin (INDO) inhibited cell migration and reduced the concentration of TNF in the exudate. Production of PGE2 or vascular permeability did not correlate with the number of cells in the pouch. Local TNF seems to play an important role in this model, particularly for leukocyte migration in the first phase of the inflammatory process. In conclusion, the air pouch seems to be a good model for studying the regulation of the early events of local inflammation, particularly the role of cytokines and cell migration

Viral Infection and Autophagy Dysregulation. The Case of HHV-6, EBV and KSHV

Human Herpes Virus-6 (HHV-6), Epstein-Barr Virus (EBV) and Kaposi Sarcoma Herpes Virus (KSHV) are viruses that share with other member of the Herpesvirus family the capacity to interfere with the autophagic process. In this paper, mainly based on the findings of our laboratory, we describe how, through different mechanisms, these viruses converge in reducing autophagy to impair DC immune function and how, by infecting and dysregulating autophagy in different cell types, they promote the pathologies associated with their infection, from the neurodegenerative diseases such Alzheimer's disease to cancer