Fast UPLC/PDA determination of squalene in Sicilian P.D.O. pistachio from Bronte: optimization of oil extraction method and analytical characterization

A fast reversed-phase UPLC method was developed for squalene determination in Sicilian pistachio samples that entry in the European register of the products with P.D.O. In the present study the SPE procedure was optimized for the squalene extraction prior to the UPLC/PDA analysis. The precision of the full analytical procedure was satisfactory and the mean recoveries were 92.8 ± 0.3 % and 96.6 ± 0.1 % for 25 and 50 mg L-1 level of addition, respectively. Selected chromatographic conditions allowed a very fast squalene determination; in fact it was well separated in ∼ 0.54 min with good resolution. Squalene was detected in all the pistachio samples analyzed and the levels ranged from 55.45 to 226.34 mg . kg-1. Comparing our results with those of other studies it emerges that squalene contents in P.D.O. Sicilian pistachio samples, generally, were higher than those measured for other samples of different geographic origins

Relative impacts of morphological alteration to shorelines and eutrophication on littoral macroinvertebrates in Mediterranean lakes

Development of effective methods for assessing the ecological status of lakes based on littoral benthic fauna has been hampered by the lack of quantitative data on the relative impacts of key pressures on the benthic community. We used variance partitioning at 126 sites belonging to 14 natural Mediterranean lakes to analyze the pure and shared effects of eutrophication, morphological alterations, microhabitat type, lake morphometry and geographic position on the littoral macroinvertebrate community. The spatial arrangement of the sampling sites was responsible for 9.1% of the total variance in littoral benthic community composition, lake morphometry accounted for 4.3% of variation, and microhabitat type accounted for 3.9%. Communities appeared to be affected primarily by morphological alterations to lake shorelines, and their impact was 2.5 times as important as that of eutrophication. The structure of littoral benthic communities was governed by processes acting at several spatial scales from region to lake scale. Thus, several pressures and the various spatial scales at which these act should be taken into account when implementing methods of assessing lake ecological condition based on littoral benthic invertebrates. Region-specific methods for subalpine and volcanic lakes might enhance the validity of assessment of results of morphological alterations and improve management of those water resources

Cooperative 3D Air Quality Assessment with Wireless Chemical Sensing Networks

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Bidirectional synaptic plasticity is driven by sex neurosteroids targeting estrogen and androgen receptors in hippocampal CA1 pyramidal neurons

Neuroactive estrogenic and androgenic steroids influence synaptic transmission, finely modulating synaptic plasticity in several brain regions including the hippocampus. While estrogens facilitate long-term potentiation (LTP), androgens are involved in the induction of long-term depression (LTD) and depotentiation (DP) of synaptic transmission. To examine sex neurosteroid-dependent LTP and LTD in single cells, patch-clamp recordings from hippocampal CA1 pyramidal neurons of male rats and selective antagonists for estrogen receptors (ERs) and androgen (AR) receptors were used. LTP induced by high-frequency stimulation (HFS) depended on activation of ERs since it was prevented by the ER antagonist ICI 182,780 in most of the neurons. Application of the selective antagonists for ERα (MPP) or ERβ (PHTPP) caused a reduction of the LTP amplitude, while these antagonists in combination, prevented LTP completely. LTP was never affected by blocking AR with the specific antagonist flutamide. Conversely, LTD and DP, elicited by low-frequency stimulation (LFS), were impeded by flutamide, but not by ICI 182,780, in most neurons. In few cells, LTD was even reverted to LTP by flutamide. Moreover, the combined application of both ER and AR antagonists completely prevented both LTP and LTD/DP in the same neuron. The current study demonstrates that the activation of ERs is necessary for inducing LTP in hippocampal pyramidal neurons, whereas the activation of ARs is required for LTD and DP. Moreover, both estrogen- and androgen-dependent LTP and LTD can be expressed in the same pyramidal neurons, suggesting that the activation of sex neurosteroids signaling pathways is responsible for bidirectional synaptic plasticity

321. Sea Urchin sns Chromatin Insulator Prevents Silencing and Positional Effect Variegation of Oncoretroviral Vectors Transgene Expression in Murine Erythroid Cell Line

Silencing and position effect are considered significant obstacles to obtain a consistent level of transgene expression in viral gene therapy. Furthermore recent studies had shown that retroviruses tend to land on active genes with the potential consequence of insertional mutagenesis. The inclusion of elements, such as chromatin insulators, capable to insulate a gene from the surrounding chromatin effects at the integration site should improve both efficacy and safety of gene therapy vectors. We have previously characterized a 265 bp insulator element, termed sns, localized at the 3' end of the early histone H2A gene of the sea urchin Paracentrotus lividus. This sequence contains three cis-acting elements (Box A, Box B, and Box C+T) all needed for the enhancer blocking activity in both sea urchin and human cells. By colony assays, in human (K562) and mouse (Mel) erythroid cell lines, we have recently demonstrated that the sns insulator displays directional enhancer-blocking activity in that it interferes with the communication between the human beta-globin enhancer (LCR) and the gamma-globin promoter. By electrophoretic mobility shift assays (EMSA) we found bindings of sns insulator with the erythroid specific GATA1 and the ubiquitous Oct1, and Sp1 transcription factors

miRNA Expression May Have Implications for Immunotherapy in PDGFRA Mutant GISTs

Gastrointestinal stromal tumors (GISTs) harboring mutations in the PDGFRA gene occur in only about 5-7% of patients. The most common PDGFRA mutation is exon 18 D842V, which is correlated with specific clinico-pathological features compared to the other PDGFRA mutated GISTs. Herein, we present a miRNA expression profile comparison of PDGFRA D842V mutant GISTs and PDGFRA with mutations other than D842V (non-D842V). miRNA expression profiling was carried out on 10 patients using a TLDA miRNA array. Then, miRNA expression was followed by bioinformatic analysis aimed at evaluating differential expression, pathway enrichment, and miRNA-mRNA networks. We highlighted 24 differentially expressed miRNAs between D842V and non-D842V GIST patients. Pathway enrichment analysis showed that deregulated miRNAs targeted genes that are mainly involved in the immune response pathways. The miRNA-mRNA networks highlighted a signature of miRNAs/mRNA that could explain the indolent behavior of the D842V mutated GIST. The results highlighted a different miRNA fingerprint in PDGFRA D842V GISTs compared to non-D842Vmutated patients, which could explain the different biological behavior of this GIST subset

Ruolo diagnostico, prognostico e predittivo di risposta del NETest nelle neoplasie neuroendocrine

Il NETest è una metodica di biologia molecolare e, in particolare, di biopsia liquida, applicata alle neoplasie neuroendocrine (NEN), che si propone come nuovo biomarcatore altamente sensibile e specifico. Il NETest consente una sorta di gene signature del tumore, definendone il profilo trascrizionale mRNA, estratto dal sangue periferico. L’applicazione pratica è nella diagnosi, dove il NETest sembra identificare anche piccoli tumori localizzati, nella definizione prognostica, con l’identificazione dei tumori con maggiore tendenza alla progressione e alla recidiva post-chirurgica, nella riposta ai trattamenti, con l’identificazione precoce di progressione nel corso di terapie anti-tumorali. A fronte di risultati iniziali estremamente promettenti, il NETest necessita di una conferma su larga scala, in ampie casistiche multicentriche

Optical modulation of excitation-contraction coupling in human-induced pluripotent stem cell-derived cardiomyocytes

Non-genetic photostimulation is a novel and rapidly growing multidisciplinary field that aims to induce light-sensitivity in living systems by exploiting exogeneous phototransducers. Here, we propose an intramembrane photoswitch, based on an azobenzene derivative (Ziapin2), for optical pacing of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). The light-mediated stimulation process has been studied by applying several techniques to detect the effect on the cell properties. In particular, we recorded changes in membrane capacitance, in membrane potential (V-m), andmodulation of intracellular Ca2+ dynamics. Finally, cell contractility was analyzed using a custom MATLAB algorithm. Photostimulation of intramembrane Ziapin2 causes a transient V-m hyperpolarization followed by a delayed depolarization and action potential firing. The observed initial electrical modulation nicely correlates with changes in Ca2+ dynamics and contraction rate. This work represents the proof of principle that Ziapin2 can modulate electrical activity and contractility in hiPSC-CMs, opening up a future development in cardiac physiology