Beneficial and Sexually Dimorphic Response to Combined HDAC Inhibitor Valproate and AMPK/SIRT1 Pathway Activator Resveratrol in the Treatment of ALS Mice

Amyotrophic lateral sclerosis (ALS) is a fatal adult-onset neurodegenerative disorder. There is no cure and current treatments fail to slow the progression of the disease. Epigenetic modulation in the acetylation state of NF-kB RelA and the histone 3 (H3) protein, involved in the development of neurodegeneration, is a drugable target for the class-I histone deacetylases (HDAC) inhibitors, entinostat or valproate, and the AMP-activated kinase (AMPK)-sirtuin 1 pathway activator, resveratrol. In this study, we demonstrated that the combination of valproate and resveratrol can restore the normal acetylation state of RelA in the SOD1(G93A) murine model of ALS, in order to obtain the neuroprotective form of NF-kB. We also investigated the sexually dimorphic development of the disease, as well as the sex-sensibility to the treatment administered. We showed that the combined drugs, which rescued AMPK activation, RelA and the histone 3 acetylation state, reduced the motor deficit and the disease pathology associated with motor neuron loss and microglial reactivity, Brain-Derived Neurotrophic Factor (BDNF) and B-cell lymphoma-extra large (Bcl-xL) level decline. Specifically, vehicle-administered males showed earlier onset and slower progression of the disease when compared to females. The treatment, administered at 50 days of life, postponed the time of onset in the male by 22 days, but not in a significant way in females. Nevertheless, in females, the drugs significantly reduced symptom severity of the later phase of the disease and prolonged the mice’s survival. Only minor beneficial effects were produced in the latter stage in males. Overall, this study shows a beneficial and sexually dimorphic response to valproate and resveratrol treatment in ALS mice.     

Campylobacter jejuni loss of culturability in aqueous microcosms and ability to resuscitate in a mouse model

Water is known as one of the main transmission routes of Campylobacter and contributes to increase the number of sporadic infections and outbreaks. Campylobacter jejuni persists in the environment, especially in water, in a viable but non-culturable (VBNC) form that is thought to be a possible cause of water-borne outbreaks. In this study, we evaluated the loss of culturability and viability of 9 C. jejuni strains of clinical origin and one ATCC reference strain when kept at 4 degrees C in artificial sea water (ASW). Culturability was measured as colony-forming units while viability was evaluated by CTC-DAPI double staining and the combined CTC-specific fluorescent antibody technique (CTC-FA). When cultured on Columbia Agar plates, strains exhibited different growth profiles which allowed to classify them into three different groups. Both techniques used to monitor the viability of the bacterial cells showed that C. jejuni strains survived in the VBNC form in the microcosms through a period lasting from 138 to 152 days. The recovery of C. jejuni VBNC forms to culturability, as evidenced by cell division, was obtained by passage in the mouse intestine. Our results indicate that C. jejuni VBNC cells were able to remain in this state for a few months and regain their culturability after in vivo passage depending on their lasting in the VBNC state, which affects the number of respiring bacteria. In fact, the resuscitation was achieved when the number of respiring bacteria became higher than 10(4) cell/ml. Therefore, a relatively high microbial titer of respiring bacteria in the VBNC state seems to be important for the resuscitation and subsequent intestinal colonisation.     

Use of duplex polymerase chain reaction (duplex-PCR) technique to identify bovine and water buffalo milk used in making mozzarella cheese

Molecular biology techniques have been used for species identification in food of animal origin in relatively recent years. A polymerase chain reaction (PCR) based method, the multiplex PCR, was recently applied to species identification in meat and meat products. It allows co-amplification of separate regions of a single gene or specific fragments, each typical of a different animal species in a single PCR reaction, using different pairs of primers in the same reaction mix. In the present paper, the duplex-PCR technique is proposed to identify bovine and water buffalo DNA in a single PCR assay in milk and mozzarella cheese (a typical Italian cheese, originally made from pure water buffalo milk). Because of its lower cost, undeclared bovine milk is added to water buffalo milk for making different kinds of mozzarella cheese. The results of this experiment indicate the applicability of this method, which showed an absolute specificity for the two species and a high sensitivity even down to low DNA concentrations (1 pg). In bovine and water buffalo mixtures of both milk and mozzarella cheese, the minimum concentration tested was 1% of bovine in water buffalo milk and water buffalo in bovine milk. The importance of the somatic cell content in raw milk is also discussed with special reference to the evaluation of mixtures (milk or cheese) of the two species.     

A face recognition system based on automatically determined facial fiducial points

In this paper, a completely automatic face recognition system is presented. The method works on color images: after having localized the face and the facial features, it determines 24 facial fiducial points, and characterizes them applying a bank of Gabor filters which extract the peculiar texture around them (jets). Recognition is realized measuring the similarity between the different jets. The system is inspired by the elastic bunch graph method, while it does no assumption on the scale, pose, and the background. Comparison with standard algorithms is presented and discussed.     

Breakdown kinetics at nanometer scale of innovative MOS devices by conductive atomic force microscopy

The breakdown (BD) kinetics of dielectrics represent a crucial issue for the reliability of microelectronics devices. In this paper, we report on an innovative and practical approach based on Conductive Atomic Force Microscopy (C-AFM) for the determination of the BD kinetics on a bare insulator surface. This technique has been applied to Pr₂O₃ films grown by Metal-Organic Chemical Vapour Deposition (MOCVD) on Si(0 0 1) and to thermally grown SiO₂ on 4H-SiC substrates. C-AFM clearly visualizes single breakdown spots under constant voltage stresses. The stress time on the C-AFM tip was varied from 1 × 10⁻³ to 1 × 10⁻¹ s. The density of BD spots, upon increasing the stress time, exhibits in both cases an exponential trend. The Weibull slope of the dielectric BD statistics has been determined by direct measurements at nanometer scale on different dielectrics having different physical thicknesses. The comparison of the Weibull slopes obtained for different dielectric thicknesses with literature data points out intrinsic and extrinsic BD events in the SiO₂/SiC system and Pr₂O₃ based layers, respectively. In the case of the SiO₂/SiC system, BD kinetics have been demonstrated to follow the percolation model, while the role of extrinsic phenomena in the BD of Pr₂O₃ films has been proved.     

Using C/X-band SAR interferometry and GNSS measurements for the Assisi landslide analysis

This work presents an analysis of the applicability of synthetic aperture radar (SAR) interferometry to landslide monitoring. This analysis was carried out by using different interferometric approaches, different spaceborne SAR data (both in the C-band and in the X-band), and in situ global navigation satellite system (GNSS) measurements. In particular, we investigated both the reliability of displacement monitoring and the issues of the cross-comparison and validation of the interferometric synthetic aperture radar (InSAR) results. The work was focused on the slow-moving landslide that affects a relevant part of the urban area of the historical town of Assisi (Italy).

A C-band ENVISAT advanced synthetic aperture radar (ENVISAT ASAR) dataset acquired between 2003 and 2010 was processed by using two different interferometric techniques, to allow cross-comparison of the obtained displacement maps. Good correspondence between the results was found, and a deeper analysis of the movement field was possible. Results were further compared to a set of GNSS measurements with a 7 year overlap with SAR data. A comparison was made for each GNSS marker with the surrounding SAR scatterers, trying to take into account local topological effects, when possible.

Further, the high-resolution X-band acquired on both ascending and descending tracks by the COSMO-SkyMed (CSK) constellation was processed. The resultant displacement fields show good agreement with C-band and GNSS measurements and a sensible increase in the density of measurements.     

Low Sulfur Amino Acid,High Polyunsaturated Fatty Acid Diet Inhibits Breast Cancer Growth

Cancer cells may acquire resistance to stress signals and reprogram metabolism to meet the energetic demands to support their high proliferation rate and avoid death. Hence, targeting nutrient dependencies of cancer cells has been suggested as a promising anti-cancer strategy. We explored the possibility of killing breast cancer (BC) cells by modifying nutrient availability. We used in vitro models of BC (MCF7 and MDA-MB-231) that were maintained with a low amount of sulfur amino acids (SAAs) and a high amount of oxidizable polyunsatured fatty acids (PUFAs). Treatment with anti-apoptotic, anti-ferroptotic and antioxidant drugs were used to determine the modality of cell death. We reproduced these conditions in vivo by feeding BC-bearing mice with a diet poor in proteins and SAAs and rich in PUFAs (LSAA/HPUFA). Western blot analysis, qPCR and histological analyses were used to assess the anti-cancer effects and the molecular pathways involved. We found that BC cells underwent oxidative damage to DNA and proteins and both apoptosis and ferroptosis were induced. Along with caspases-mediated PARP1 cleavage, we found a lowering of the GSH-GPX4 system and an increase of lipid peroxides. A LSAA/HPUFA diet reduced tumor mass and its vascularization and immune cell infiltration, and induced apoptosis and ferroptotic hallmarks. Furthermore, mitochondrial mass was found to be increased, and the buffering of mitochondrial reactive oxygen species limited GPX4 reduction and DNA damage. Our results suggest that administration of custom diets, targeting the dependency of cancer cells on certain nutrients, can represent a promising complementary option for anti-cancer therapy.     

Oil viscosity measurement by ultrasonic reflectance

Pulses of shear-mode ultrasound (center frequency 10 MHz) are reflected from the surface of a series of vegetable and synthetic high-viscosity calibration oils at a range of temperatures (5–50°C). For all samples and temperatures there is a single negative correlation between the magnitude of the echo from the interface between the delay-line and the sample and the viscosity of the sample. Similar experiments with longitudinal ultrasonic waves show the amount of sound reflected decreased with increasing viscosity, but there is no single correlation for all samples.     

Identification and functional traits of lactic acid bacteria isolated from Ciauscolo salami produced in Central Italy

Lactic acid bacteria (LAB) from Ciauscolo salami produced in Marche Region of Central Italy, and LAB strains belonging to our laboratory collection were examined for their capability to survive at low pH and bile, to adhere to Caco-2 cells, and for antibiotic resistance. LAB from Ciauscolo were identified by ARDRA and RAPD-PCR. Our study showed that all LAB strains had good adaptation to gastric juice and moderate tolerance to bile. The adhesiveness was variable among strains but significantly lower in LAB from food. Antibiotic resistance was broadly spread among food strains, with level of resistance exceeding 15% for all the antibiotics tested. The resistance determinants erm(B) and tet(M) were found in nine strains of food origin (21.4%) while tet(L) in one strain of our collection (5%). Our work suggests that fermented foods are valuable sources of bacterial strains with functional traits of intestinal lactobacilli. These bacteria may be further studied for their use in probiotic applications.     

Potential Effect of CD271 on Human Mesenchymal Stromal Cell Proliferation and Differentiation

The Low-Affinity Nerve Growth Factor Receptor (LNGFR), also known as CD271, is a member of the tumor necrosis factor receptor superfamily. The CD271 cell surface marker defines a subset of multipotential mesenchymal stromal cells and may be used to isolate and enrich cells derived from bone marrow aspirate. In this study, we compare the proliferative and differentiation potentials of CD271⁺ and CD271⁻ mesenchymal stromal cells. Mesenchymal stromal cells were isolated from bone marrow aspirate and adipose tissue by plastic adherence and positive selection. The proliferation and differentiation potentials of CD271⁺ and CD271⁻ mesenchymal stromal cells were assessed by inducing osteogenic, adipogenic and chondrogenic in vitro differentiation. Compared to CD271⁺, CD271⁻ mesenchymal stromal cells showed a lower proliferation rate and a decreased ability to give rise to osteocytes, adipocytes and chondrocytes. Furthermore, we observed that CD271⁺ mesenchymal stromal cells isolated from adipose tissue displayed a higher efficiency of proliferation and trilineage differentiation compared to CD271⁺ mesenchymal stromal cells isolated from bone marrow samples, although the CD271 expression levels were comparable. In conclusion, these data show that both the presence of CD271 antigen and the source of mesenchymal stromal cells represent important factors in determining the ability of the cells to proliferate and differentiate.