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The purpose of the present study was to evaluate the impact of long-term honey ingestion on metabolic disorders and neurodegeneration in mice fed a high-fat diet (HFD). Three groups of mice were fed with a standard diet (STD), HFD or HFD supplemented with honey (HFD-H) for 16 weeks. Biochemical, histological, Western blotting, RT-PCR and Profiler PCR array were performed to assess metabolic parameters, peripheral and central insulin resistance and neurodegeneration. Daily honey intake prevented the HFD-induced glucose dysmetabolism. In fact, it reduced plasma fasting glucose, insulin and leptin concentrations and increased adiponectin levels. It improved glucose tolerance, insulin sensitivity and HOMA index without affecting plasma lipid concentration. HFD mice showed a significantly higher number of apoptotic nuclei in the superficial and deep cerebral cortex, upregulation of Fas-L, Bim and P27 (neuronal pro-apoptotic markers) and downregulation of Bcl-2 and BDNF (anti-apoptotic factors) in comparison with STD- and HFD-H mice, providing evidence for honey neuroprotective effects. PCR-array analysis showed that long-term honey intake increased the expression of genes involved in insulin sensitivity and decreased genes involved in neuroinflammation or lipogenesis, suggesting improvement of central insulin resistance. The expressions of p-AKT and p-GSK3 in HFD-H mice, which were decreased and increased, respectively, in HFD mouse brain, index of central insulin resistance, were similar to STD animals supporting the ability of regular honey intake to protect brain neurons from insulin resistance. In conclusion, the present results provide evidence for the beneficial preventative impact of regular honey ingestion on neuronal damage caused by HFD.

Motivation: An increase in human life span is expected to double in the next 30 years. As the elderly population expands, the prevalence of neurodegenerative diseases such as Alzheimer's disease (AD) or other forms of dementia is likely increasing, inducing consequently profound economic and social implications. Thirty-five million people are now considered to be affected by AD and this number is expected to double in the next few decades. Consequently, AD is now considered a very serious public health problem mainly for the direct and indirect costs of their care. One of the histopathological hallmarks of AD is the amyloid plaques formation in the brain. The plaques are mainly constituted by aggregates of A? peptide, a peptide generated by a proteolytic cleavage of the amyloid precursor protein (APP). However, the early events in the onset of the pathology are yet to be fully explained and the mechanisms through which A? triggers the cascade of events leading to AD are unknown. Alterations in proteomic pathway have been described in many neurodegenerative diseases, including in AD. The aim of this study was identified potential biomarkers involved in early AD stage by using proteomic approach and bioinformatic tools. For this purpose, we experimentally mimed AD early stage conditions by using neuronal cells treated for short time with A? peptide. Methods: LAN5 neuroblastoma cells were incubated without (control) or with A? peptide for a short time (1 hour) at 37°C. Proteins obtained from these cells were analyzed with a shotgun proteomics approach by LTQ-Orbitrap-MS followed by label-free quantitation. Significantly up- and down-regulated proteins were identified by volcano plot STRING was used to visualize protein-protein interaction between the proteins identified as modulated. DAVID and REViGO were used for GO annotation. LAN5 protein expression after A? treatment was analyzed by Western blot incubated with specific antibodies. Results: An in vitro AD early stage was simulate by treating LAN5 cells with A? for short time. Protein extracted from untreated and treated cells were subjected to shotgun proteomic analysis followed by bioinformatics approach. A volcano plot was built from the mass spectrometry data for visualizing significantly modulated proteins (p < 0. 05 and log2- fold change 0.5). A total of 159 proteins were found to be up-regulated and 88 to be down-regulated relative to the control. By STRING and GO annotation two protein-protein interaction networks were identified. The first network contained proteins related to rRNA metabolism and the second one consisted mainly of proteins related to RNA metabolism, and many with the splicing process. The pathway enrichment identified using KEGG mainly showed up-regulation of the lysosomal pathway and down regulation of the spliceosomal pathway. To confirm that A? inhibits expression of proteins involved in the spliceosomal pathway the modulation of expression of SmB /B'/N and PRP6, two ubiquitous proteins of the spliceosome, was analyzed. By Western blot incubated with specific antibodies we confirmed down regulation of these proteins in a time-dependent manner. In conclusion, this study provides evidence that a short exposure of LAN5 with A? can be considered a model to mimic early AD stages. Several up- and down-regulated proteins involved in different dysfunction occurring in AD have been found as a consequence of A? stimulation. In particular, down-regulation of spliceosome components suggests that the modification of the splicing machinery is a key event in early AD and the proteins involved can be utilized as potential biomarkers.

Background. Multiple myeloma (MM) is characterized by inherited susceptibility and marked genetic instability. Polymorphisms in multiple absorption, distribution, metabolism and excretion (ADME) genes involved in metabolism of carcinogenic agents may contribute to cancer risk through a change in the gene expression and/or reduction of their products' activity. We investigated the relationship between MM susceptibility and xenobiotic metabolism by the drug-metabolizing enzyme and transporter (DMET(TM)) microarray Affymetrix platform, by genotyping 1936 genetic variants across 231 ADME genes by interrogating 1931 SNPs and 5 CNVs in Linkage disequilibrium (LD) in 65 MM patients matched with a dataset of 59 Caucasian HapMap-related healthy controls in a case control study. Methods. Genomic DNA, extracted from peripheral blood cells, was genotyped by Affymetrix DMET Plus microarray and the genotyping profiles were generated by DMET Console software® algorithm. Genotype frequencies and association analysis were analyzed by DMET-Analyzer software. Results of potential interest were limited to those in which the p-value was

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The aim of this study is to investigate the neurobiology of stress/emotionality, creating a multidisciplinary assessment model, which can help to provide psychological and physiological responses depending on the genetic background related to sport performances, social closeness and performance anxiety management in team sports. We enrolled 20 female volleyball players aged 13 ± 1 years old played in two different teams during a regional championship final. Saliva collection was carried out before and after the match. In order to evaluate the neuroendocrine effectors involved in stress and performance, we analyzed cortisol and progesterone levels through Elisa standard kit as well as HSP70 and amylase activity as stress-induced markers. As concern the psychometric assesment, we administrated he CSAI-2 test, Closeness Generating Procedure and STAI test. Genomic DNA was isolated from saliva cells using a QIAamp saliva kit according to the manufacturer’s protocols. The SNP of the 5-HTTLPR, BDNF, DRD4 were analyzed. The results of the T-test performed on the total results showed a statistically significant relationship (p < 0.05) in cortisol levels pre and post match, as well between amylase and HP70 according to the genetic background. The analysis performed using just post match samples show a negative correlation between social closeness, cortisol and progesterone levels, with p < 0.010 for progesterone vs social closeness and p < 0.012 for cortisol vs social closeness. About the winner teams and the looser teams, there is a negative correlation between pre match cortisol levels and performance anxiety (p < 0.042).

A growing body of evidence shows that Insulin, Insulin Receptor (IR) and IR signaling are involved in brain cognitive functions and their dysfunction is implicated in Alzheimer’s disease (AD) degeneration. Thus, administration of insulin could be a strategy for AD treatment. For this aim we have designed, synthesized and characterized a nanogel system (NG) to deliver insulin to the brain, as a tool for the development of a new therapy for AD. A carboxyl-functionalized poly(N-vinyl pyrrolidone) nanogel system produced by ionizing radiation was chosen as substrate for the covalent attachment of insulin or fluorescent molecules relevant for its characterization. Biocompatibility of the naked carrier was demonstrated by absence of cytotoxicity, oxidative stress and mitochondrial dysfunction. Hemocompatibility was demonstrated by hemolysis, coagulation time, leukocyte proliferation and inflammatory response tests. By immonufluorescence measurements we demonstrated that insulin conjugated to the NG (NG-In) is preotected by protease degradation and is able to bind and activate insulin receptor bringing to trigger the insulin signalling via AKT activation. Moreover, to provide consistent evidence on the functionality of the conjugated insulin on the glucose levels, the effect of NG-In was tested in mice demonstrating that plasma glucose levels was reduced. Neuroprotection of NG-In against dysfunction induced by amyloid β, a peptide mainly involved in AD, was verified. Finally, the potential of NG-In to be efficiently transported across the Blood Brain Barrier was demonstrated by using an in vitro system. All together these results indicated that the synthesized NG-In was a suitable vehicle system for insulin delivery in biomedicine and a very promising tool to develop new therapies for neurodegenerative diseases. The research eas supported by MIUR, FLAGSHIP PROJECT NanoMAX.

I nanogeli sono nanoparticelle polimeriche reticolate idrofile. La struttura polimerica del nanogelo permette la coniugazione di molecole che possano esplicare azioni terapeutiche, diagnostiche e di indirizzamento attivo su siti bersaglio. Le caratteristiche di flessibilità e conformabilità del nanogelo, e delle sue maglie, ne favoriscono l’attraversamento delle barriere biologiche, l’interazione di eventuali ligandi esposti sulla superfice con i recettori target, un’azione protettiva nei confronti del principio attivo incorporato. L’irraggiamento con radiazioni ionizzanti di soluzioni acquose di polimeri è una metodologia sintetica interamente “water-based”, che garantisce rese elevate e che può conferire sterilità al prodotto. [1] Lo studio presentato illustra la possibilità di produrre nanogeli a base di polivinilpirrolidone con le caratteristiche desiderate in termini di dimensioni e con gruppi funzionali disponibili per reazioni di coniugazione con l’insulina. L’obiettivo è quello di realizzare un nanovettore in grado di veicolare l’insulina al cervello attraverso la mucosa nasale. Studi recenti hanno dimostrato che l’insulina è in grado di ridurre la tossicità del peptide beta amiloide (A, coinvolto nella patogenesi della malattia di Alzheimer (MA), e di esplicare una funzione neuroprotettiva. [2, 3] Sono state studiate le caratteristiche dimensionali del nanosistema, la biocompatibilità e la funzione di protezione dalla tossicità indotta da Anei confronti di cellule di neuroblastoma.

The aim of this study is to investigate the neurobiology of stress/emotionality, creating a multidisciplinary assessment model, which can help to provide psychological and physiological responses depending on the genetic background related to sport performances, social closeness and performance anxiety management in team sports. We enrolled 20 female volleyball players aged 13 ± 1 years old played in two different teams during a regional championship final. Saliva collection was carried out before and after the match. In order to evaluate the neuroendocrine effectors involved in stress and performance, we analyzed cortisol and progesterone levels through Elisa standard kit as well as HSP70 and amylase activity as stress-induced markers. As concern the psychometric assesment, we administrated he CSAI-2 test, Closeness Generating Procedure and STAI test. Genomic DNA was isolated from saliva cells using a QIAamp saliva kit according to the manufacturer’s protocols. The SNP of the 5-HTTLPR, BDNF, DRD4 were analyzed. The results of the T-test performed on the total results showed a statistically significant relationship (p < 0.05) in cortisol levels pre and post match, as well between amylase and HP70 according to the genetic background. The analysis performed using just post match samples show a negative correlation between social closeness, cortisol and progesterone levels, with p < 0.010 for progesterone vs social closeness and p < 0.012 for cortisol vs social closeness. About the winner teams and the looser teams, there is a negative correlation between pre match cortisol levels and performance anxiety (p < 0.042).

Nanogels are nanoscalar polymeric networks, characterized by a dynamic internal structure and a flexible adaptive shape. When they are used as drug carriers, their flexibility and shape changing ability can facilitate the bypass of biological barriers, ensure protection of the payload and enable interaction of any attached ligand with its receptors. Poly(N-vinyl pyrrolidone)-co-acrylic acid nanogels (NGs) have been produced by ebeam irradiation of diluted aqueous solutions of a water-soluble polymer in the presence of a small amount of acrylic acid. In particular, industrial accelerators and the typical set-ups and doses applied for sterilization have been used. [1, 2] Particle size, molecular weight and functionalization can be controlled by a proper selection of polymer concentration and irradiation parameters, such as dose-rate and total dose imparted [3]. In recent years, a growing body of evidence has linked insulin resistance and insulin action to Alzheimer’s Disease (AD), a condition also referred to as Type 3 Diabetes (T3D) [4,5]. Recently, it has been demonstrated that insulin is capable of reducing toxicity induced by A oligomers, the protein mainly involved in AD onset, by inhibition of the intrinsic apoptotic pathway [6]. Moreover, activation of insulin signaling provides a neuroprotective mechanism to counteract oxidative stress, mitochondrial damage and neurodegeneration triggered by A oligomers in neuroblastoma cells [7]. In the present work NGs have been conjugated to insulin (NG-In) to defend the hormone from enzymatic degradation, facilitate its crossing through a blood brain barrier model and protect the LAN5 cells from the damages induced by A oligomers addiction.

Introduction: Saliva collection and analysis is quickly becoming a useful and non-invasive tool for the evaluation of sport biomarkers. The aim of this study is to create a multidisciplinary assessment model, which can help to provide psychological and physiological responses, related to sport performances, social closeness and performance anxiety management in team sports. Materials and methods: We enrolled in our research 26 female volleyball players aged 13 ± 1 years old of three different teams (T1: 12 players; T2: 9 players; T3: 5 players). Saliva collection was carried out before and after the match for every team. Then we analyzed cortisol and progesterone concentrations through Elisa standard kits. Results: The results of the T-test performed on the total results showed a statistically significant relationship (p < 0.05) in cortisol levels pre and post match: in fact, it has been shown a statistical significant decrease (p < 0.001). The analysis performed using just samples post match shows a negative correlation between social closeness, cortisol and progesterone levels, with p < 0.010 for progesterone vs social closeness and p < 0.012 for cortisol vs social closeness, which indicates that increasing of one of the two hormones reduces relationship. About the winner teams and the looser teams, there is a negative correlation between pre match cortisol levels and performance anxiety (p < 0.042).

Nanogels (NGs) have a great potential in the development of “smart” nanocarriers for (bio)molecular drugs and contrast agent for bioimaging. They are formed by physically or chemically crosslinked polymer networks, characterized by a large and flexible surface available for multivalent bioconjugations. NGs can be produced with high yields and through-puts by pulsed electron-beam irradiation of dilute aqueous solutions of water-soluble biocompatible polymers. In this work, a carboxyl functionalized nanogel system (NG), generated by pulsed e-beam irradiation of a semi-dilute poly(N-vinyl pyrrolidone) (PVP) aqueous solution in the presence of acrylic acid, with an average diameter in the 60-70 nm range (PDI

Insulin, a metabolic hormone involved in glucose metabolism, plays also a neuroprotective role in the central nervous system being able to revert the cytotoxic processes induced by Aβ42,a peptide involved in Alzheimer's disease. To reach the brain insulin have to across the BBB therefore an additional delivery strategy results to be necessary. For these aim we performed an insulin conjugated nanogels (NGs-In). Nanogels (NGs) have a great potential in the development of “smart” nanocarriers for (bio)molecular drugs and contrast agent for bioimaging. They are formed by physically or chemically crosslinked polymer networks, characterized by a large and flexible surface available for multivalent bioconjugations. NGs can be produced with high yields and through-puts by pulsed electron-beam irradiation of dilute aqueous solutions of water-soluble biocompatible polymers. In this work, a carboxyl functionalized nanogel system (NG), generated by pulsed e-beam irradiation of a semi-dilute poly(N-vinyl pyrrolidone) (PVP) aqueous solution in the presence of acrylic acid, with an average diameter in the 60-70 nm range (PDI

A growing body of evidence shows that Insulin, Insulin Receptor (IR) and IR signaling are involved in brain cognitive functions and their dysfunction is implicated in neuronal degeneration associated with Alzheimer's disease (AD). Thus, the administration of insulin to the brain could be a strategy for the prevention and treatment of AD disease. With this aim, we have designed, synthesized and characterized a nanogel system (NG) that can be used as substrate for the conjugation of insulin and/or fluorescent molecules relevant for their characterization. In particular, a carboxyl-functionalized poly(N-vinyl pyrrolidone) nanogel system, has been produced by ionizing radiation starting from the polymeric aqueous solution and four insulin molecules per nanogel have been irreversibly attached. Absence of cytotoxicity, oxidative stress and mitochondrial dysfunction support the biocompatibility of the “naked” nanogels. Their hemocompatibility has been demonstrated by hemolysis, coagulation time, leukocyte proliferation and inflammatory response tests. By fluorescence measurements we have demonstrated that the insulin conjugated to the NG (NG-In) is protected by protease degradation and is able to bind and activate insulin receptor, thus triggering insulin signaling via AKT activation. In order to provide more significant evidence of the functionality of the insulin conjugated to the nanoparticles at the body level, the effect of NG-In in reducing the plasma glucose levels in mice has been demonstrated. Neuroprotection of NG-In against dysfunction induced by amyloid β, a peptide mainly involved in AD, has been also verified. Finally, the potential of NG-In to be efficiently transported across the Blood Brain Barrier has been demonstrated in vitro. All together, these results indicate that the synthesized NG-In is a suitable vehicle system for insulin deliver and a very promising tool to develop new therapies for neurodegenerative diseases

$\textbf{A perovskite LED with a perovskite film}$ treated under optimum thermal annealing conditions exhibits a significantly enhanced long-term stability with full coverage of the green electroluminescence emission due to the highly uniform morphology of the perovskite film.

Mitochondria are dynamic ATP-generating organelle which contribute to many cellular functions including bioenergetic processes. intracellular calcium regulation, alteration of reduction-oxidation potential of cells, free radical scavenging, and activation of caspase-mediated celi death. Many lines of evidence suggest that mitochondria have a central role in ageing-related neurodegenerative diseases. Mitochondrial functions can be negatively affected by Amyloid Beta-peptide (ABeta), an important component in Alzheimer's disease (AD) pathogenesis, and ABeta can interact with mitochondria and cause mitochondrial dysfunction. One of the most accepted hypotheses for AD onset implicates that mitochondrial dysfunction and oxidative stress are one of the primary events in the insurgence of the pathology. To examine structural and functional mitochondrial changes in presence of Abeta, we utilized a proteomic approach by using an Orbitrap-based mass spectrometer that use a new physical principle to obtain high resolution. The Orbitrap is an advanced instrume nt for shotgun proteomics combining high-resolving power and mass accuracy that leads to higher quality datasets and reduction of false positive peptide identifications. For this aim we used neuroblastoma LAN5 cells treated without or with ABeta and submitted to cellular fractionation. Mitochondrial fractions were separated and ulilized to extract protcins to be submitted to proteomic analysis by using an LTQ-Orbitrap Velos mass spectrometer, followed by tandem mass spectra (MS/MS) examination. Proteome Discoverer platform interfaced with an in-house Mascot server was used for data analyzing and protein identification. together with database UniProtKB/Swiss-Prot. The spectral count (SpC) log ratio was used to express fold-change between different conditions. Using the program lngenuity Systems, lnc, we evidenced the involvement of some proteins in specific networks. About 500 mitochondrial proteins were identified and some of them were up- or down-regulated in the treated sample. Some of thcse proteins are involved in mitochondrial oxidative stress, bioenergetics, biogenesis, trafficking, itochondrial permeability transition pore (mPTP) formation and mitochondrial protein interaction, all mechanisms whose alteration leads to mitochondrial ysfunction.

In the recent years a growing body of evidence links insulin resistance and insulin action to neurodegenerative diseases, especially Alzheimer’s disease (AD). The importance of insulin in ageing as well as its role in cognition and other aspects of normal brain functions are well established. The hippocampus and cerebral cortex-distributed insulin and insulin receptor (IR) have been shown to be involved in brain cognitive functions. Conversely, deterioration of IR signaling is involved in agingrelated brain degeneration such as in AD and cognitive impairment in type 2 diabetes patients. Insulin administration, while maintaining euglycemia, improves memory in both healthy adults and Alzheimer’s disease patients. In the present review, some common links between AD and type 2 diabetes are presented. Furthermore, several biochemical aspects existing in both pathologies are highlighted.

Aging is a complex process that negatively impacts the development of the different systems and its ability to function. Moreover, the Aging rate in humans is not the same, principally due to genetic heterogeneity and environmental factors. The aging rate is measured as the decline of functional capacity and stress resistance. Therefore, several attempts have been made to analyse the individual age, ( so-called biological age) compared to chronological age. The biomarkers of aging are age-related body function or composition, these markers aim to assess the biological age and predict the onset of age-related diseases and/or residual lifetime. Such biomarkers should help in one hand to characterise the biological age and on the other hand to identify individuals at high risk of developing age-associated diseases or disabilities. Unfortunately, most of the markers under discussion are related to age-related diseases rather than to age, so none of these markers discussed in literature is a true biomarker of aging. Hence, we discuss some disease-related biomarkers useful for a better understanding of aging and the development of new strategies to counteract it, essential for improving the quality of life of the elderly population.

Distribution of T29C TGFb1 gene polymorphism was analysed in 260 hypertensive and 134 normotensive subjects. Circulating TGFb1 and procollagen type III levels, microalbuminuria, left ventricular geometry and function were evaluated in all the hypertensives subgrouped according to T29C TGFb1 gene polymorphism. Circulating TGFb1by ELISA technique, procollagen type III by a specific radioimmunoassay, microalbuminuria by radioimmunoassay, left ventricular geometry and function by echocardiography were determined. All groups were comparable for gender, age and sex. Regarding T29C TGFb1 gene polymorphism, prevalence of TC or CC genotypes was significantly (pv0.05) higher in hypertensives than normotensives. TC and CC hypertensives were characterized by a higher prevalence of subjects with microalbuminuria (pv0.001 TC vs TT; pv0.05 CC vs TT), left ventricular hypertrophy (pv0.01 TC and CC vs TT), and by increased levels of procollagen type III (pv0.05 TC and CC vs TT). TC hypertensives were also characterized by a significant increase (pv0.05) of LVM and LVM/h and of urinary albumin excretion (pv0.05) values than those detectable in TT hypertensives. Our data suggest that T29C TGFb1 gene polymorphism was associated to clinical characteristics suitable to recognize hypertensives with a higher severity of hypertension.