miRNA Expression Profiling in Subcutaneous Adipose Tissue of Monozygotic Twins Discordant for HIV Infection: Validation of Differentially Expressed miRNA and Bioinformatic Analysis

Combined AntiRetroviral Treatments (cARTs) used for HIV infection may result in varied metabolic complications, which in some cases, may be related to patient genetic factors, particularly microRNAs. The use of monozygotic twins, differing only for HIV infection, presents a unique and powerful model for the controlled analysis of potential alterations of miRNAs regulation consequent to cART treatment. Profiling of 2578 mature miRNA in the subcutaneous (SC) adipose tissue and plasma of monozygotic twins was investigated by the GeneChip^® miRNA 4.1 array. Real-time PCR and ddPCR experiments were performed in order to validate differentially expressed miRNAs. Target genes of deregulated miRNAs were predicted by the miRDB database (prediction score > 70) and enrichment analysis was carried out with g:Profiler. Processes in SC adipose tissue most greatly affected by miRNA up-regulation included (i) macromolecular metabolic processes, (ii) regulation of neurogenesis, and (iii) protein phosphorylation. Furthermore, KEGG analysis revealed miRNA up-regulation involvement in (i) insulin signaling pathways, (ii) neurotrophin signaling pathways, and (iii) pancreatic cancer. By contrast, miRNA up-regulation in plasma was involved in (i) melanoma, (ii) p53 signaling pathways, and (iii) focal adhesion. Our findings suggest a mechanism that may increase the predisposition of HIV⁺ patients to insulin resistance and cancer.     

Online Portable Microcantilever Biosensors for Salmonella enterica Serotype Enteritidis Detection

The micro- and nano-technologies coupled with a deep knowledge of organic/inorganic interfaces guarantee an exceptional sensitivity and specificity of the sensor, while the lab-on-a-chip platform reduces assay times and limits sampling and/or sample preparation, providing compact and portable objects. Therefore, the development of innovative biosensors such as antibody-immobilized microcantilevers can overcome the evident limits of nowadays technologies, such as time consuming, expensiveness, difficult automation, low sensitivity, accuracy, and precision for quantitative methods. The present study proposes two device designs for the detection of food pathogens, exploiting an antibody-immobilized microcantilever biosensors, a novel class of mass detectors. For the first one, we integrated the mechanical sensors on a microfluidic platform (lab-on-a-chip) to perform online analysis, directly in liquid environment. We showed that our portable biosensors could easily detect the presence of pathogenic bacteria such as Salmonella enterica serotype enteritidis in concentration 10⁵ cfu/mL in just 40 min, without any enrichment and/or sample preparation. To increase the mass sensitivity of our analysis, we also fabricated microstructures optimized for vibrating in vacuum environment. Using a dip-and-dry technique, we showed that, in such configuration, the experimental limit of detection is as low as 10³ cfu/mL. Due to the extremely small volumes needed, our biosensors operating in vacuum have the potentiality of detecting the presence or absence of a single cell.     

External Shape Differences between Sympatric Populations of Commercial Clams <Emphasis Type="Italic">Tapes decussatus</Emphasis> and <Emphasis Type="Italic">T. philippinarum</Emphasis>

The actual Italian production of clams is chiefly sustained by the native Tapes decussatus and the fortuitously imported Tapes philippinarum. Both species are commercialized as “Vongola verace”, but the commercial value of T. philippinarum is lower. The discrimination of species by sight is usually difficult and it cannot be done by observation based on shell morphology but only when animals open their valves hence displaying the two siphons. In this study, we propose a new, noninvasive method to discriminate individuals of both species based on the analysis of the external shape of their shells. Accordingly, in sympatric populations at two sites of the Po river outlet, we have chosen individuals (63 for T. decussatus and 57 for T. philippinarum) of comparable commercial size for which a certain genetic discrimination was previously done. Pictures of the left side valve were taken for all specimens. Their profiles were analyzed with the elliptic Fourier analysis. The mean outline for each species was graphically extracted. The coefficients of the harmonic equations were analyzed by multivariate classification (partial least squares discriminant analysis [PLSDA]). Results showed a high percentage of correct classification of individuals of both species (96.6%). Contour analysis reflected the overall shell shape and thus identified morphological aspects that were difficult to recognize and quantify in sight. The high percentage of correct classifications obtained by combining the analysis of elliptic Fourier harmonics with PLSDA demonstrated the feasibility of this method to discriminate species with a high level of resemblance.     

Synthesis of poly(N‐isopropylacrylamide) by distillation precipitation polymerization and quantitative grafting on mesoporous silica

In this work, syntheses of thermoresponsive poly(N‐isopropylacrylamide) (PNIPAM) with different molecular weights were carried out in ethanol by distillation precipitation polymerization (DPP) technique. The synthesized polymers were fully characterized by attenuated total reflection Fourier‐transform infrared (ATR‐FTIR) spectroscopy, nuclear magnetic resonance spectroscopy, and size exclusion chromatography techniques. The lower critical solution temperatures of the polymers were determined with differential scanning calorimetry. A simple and versatile method for the in situ synthesis and grafting of PNIPAM on mesoporus silica nanoparticles (MSNs) with improved control over quantitative grafting is devised. The PNIPAM grafted MSNs were characterized with ATR‐FTIR, thermogravimetric analysis, transmission electron microscopy, and dynamic light scattering analyses. From the results obtained it is showed that quantitative grafting of PNIPAM on MSNs from 1 to 20% by weight can be tuned by manipulating the in situ DPP reaction conditions. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 44181.     

The Expression of Active CD11b Monocytes in Blood and Disease Progression in Amyotrophic Lateral Sclerosis

Monocytes expressing the inflammation suppressing active CD11b, a beta2 integrin, may regulate neuroinflammation and modify clinical outcomes in amyotrophic lateral sclerosis (ALS). In this single site, retrospective study, peripheral blood mononuclear cells from 38 individuals living with ALS and 20 non-neurological controls (NNC) were investigated using flow cytometry to study active CD11b integrin classical (CM), intermediate (IM) and non-classical (NCM) monocytes during ALS progression. Seventeen ALS participants were sampled at the baseline (V1) and at two additional time points (V2 and V3) for longitudinal analysis. Active CD11b+ CM frequencies increased steeply between the baseline and V3 (ANOVA repeated measurement, p < 0.001), and the V2/V1 ratio negatively correlated with the disease progression rate, similar to higher frequencies of active CD11b+ NCM at the baseline (R = −0.6567; p = 0.0031 and R = 0.3862; p = 0.0168, respectively). CD11b NCM, clinical covariates and neurofilament light-chain plasma concentration at the baseline predicted shorter survival in a multivariable and univariate analysis (CD11b NCM—HR: 1.05, CI: 1.01–1.11, p = 0.013. Log rank: above median: 43 months and below median: 21.22 months; p = 0.0022). Blood samples with the highest frequencies of active CD11b+ IM and NCM contained the lowest concentrations of soluble CD11b. Our preliminary data suggest that the levels of active CD11b+ monocytes and NCM in the blood predict different clinical outcomes in ALS.     

Cancer-Related Cachexia: The Vicious Circle between Inflammatory Cytokines,Skeletal Muscle,Lipid Metabolism and the Possible Role of Physical Training

Cachexia is a multifactorial and multi-organ syndrome that is a major cause of morbidity and mortality in late-stage chronic diseases. The main clinical features of cancer-related cachexia are chronic inflammation, wasting of skeletal muscle and adipose tissue, insulin resistance, anorexia, and impaired myogenesis. A multimodal treatment has been suggested to approach the multifactorial genesis of cachexia. In this context, physical exercise has been found to have a general effect on maintaining homeostasis in a healthy life, involving multiple organs and their metabolism. The purpose of this review is to present the evidence for the relationship between inflammatory cytokines, skeletal muscle, and fat metabolism and the potential role of exercise training in breaking the vicious circle of this impaired tissue cross-talk. Due to the wide-ranging effects of exercise training, from the body to the behavior and cognition of the individual, it seems to be able to improve the quality of life in this syndrome. Therefore, studying the molecular effects of physical exercise could provide important information about the interactions between organs and the systemic mediators involved in the overall homeostasis of the body.     

CD8+ T Cell Senescence: Lights and Shadows in Viral Infections,Autoimmune Disorders and Cancer

CD8⁺ T lymphocytes are a heterogeneous class of cells that play a crucial role in the adaptive immune response against pathogens and cancer. During their lifetime, they acquire cytotoxic functions to ensure the clearance of infected or transformed cells and, in addition, they turn into memory lymphocytes, thus providing a long-term protection. During ageing, the thymic involution causes a reduction of circulating T cells and an enrichment of memory cells, partially explaining the lowering of the response towards novel antigens with implications in vaccine efficacy. Moreover, the persistent stimulation by several antigens throughout life favors the switching of CD8⁺ T cells towards a senescent phenotype contributing to a low-grade inflammation that is a major component of several ageing-related diseases. In genetically predisposed young people, an immunological stress caused by viral infections (e.g., HIV, CMV, SARS-CoV-2), autoimmune disorders or tumor microenvironment (TME) could mimic the ageing status with the consequent acceleration of T cell senescence. This, in turn, exacerbates the inflamed conditions with dramatic effects on the clinical progression of the disease. A better characterization of the phenotype as well as the functions of senescent CD8⁺ T cells can be pivotal to prevent age-related diseases, to improve vaccine strategies and, possibly, immunotherapies in autoimmune diseases and cancer.     

Microdroplet-based multiplex PCR on chip to detect foodborne bacteria producing biogenic amines

The development of fast, reliable and culture-independent molecular tools to detect bacteria producing biogenic amines deserves the attention of research and ultimately of the food industry in order to protect consumers' health. Here we present the application of a simple, low-cost, fast and sensitive method to perform microdroplet-based multiplex PCR, directly on a food matrix, for the simultaneous detection of bacterial genes involved in biogenic amine biosynthesis. After inoculating wine with Lactobacillus brevis IOEB 9809, cell lysis and DNA amplification are performed in one single step, without preliminary nucleic acid extraction or purification treatments. The assay is performed in about 30 min, requiring 150 nL of starting sample and it enables the detection of down to 15 bacterial cells. With respect to traditional culture techniques, the speed, the simplicity and the cheapness of this procedure allow an effective monitoring of microbial cells during food-making and processing.     

Modification of carboxyl groups of poly(ethylene-co-acrylic acid) via facile wet chemistry method: A kinetic study

In this work, we present the results of a kinetic study of the surface modification of the carboxyl group of poly(ethylene-co-acrylic acid) into acyl chloride. Reaction temperature and time, concentration of the reactants, kind of agitation have been changed in order to understand their effect on the conversion. A study on the penetration of the reaction inside the sample bulk was also performed. The results indicate that, when short reaction times are adopted, the reaction limits to the outer layers of the samples and, in this case, on increasing the temperature, the concentration of the reactants and the stirring allows having a faster kinetic. At longer reaction times, the reaction penetrates deep inside the sample (several tens of microns) and the influence of the above mentioned variables on the kinetic is very low.The results, obtained by direct FTIR, ATR-FTIR, SEM–EDX and contact angle measurements, were confirmed and corroborated by theoretical estimations.     

β-Cyclodextrin Inclusion Complex to Improve Physicochemical Properties of Pipemidic Acid: Characterization and Bioactivity Evaluation

The aptitude of cyclodextrins (CDs) to form host-guest complexes has prompted an increase in the development of new drug formulations. In this study, the inclusion complexes of pipemidic acid (HPPA), a therapeutic agent for urinary tract infections, with native β-CD were prepared in solid state by kneading method and confirmed by FT-IR and ¹H NMR. The inclusion complex formation was also characterized in aqueous solution at different pH via UV-Vis titration and phase solubility studies obtaining the stability constant. The 1:1 stoichiometry was established by a Job plot and the inclusion mechanism was clarified using docking experiments. Finally, the antibacterial activity of HPPA and its inclusion complex was tested on P. aeruginosa, E. coli and S. aureus to determine the respective EC_50s and EC_90s. The results showed that the antibacterial activity of HPPA:β-CD against E. coli and S. aureus is higher than that of HPPA. Furthermore, HPPA and HPPA:β-CD, tested on human hepatoblastoma HepG2 and MCF-7 cell lines by MTT assay, exhibited, for the first time, antitumor activities, and the complex revealed a higher activity than that of HPPA. The use of β-CD allows an increase in the aqueous solubility of the drug, its bioavailability and then its bioactivity.