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Previous work, using fluorescent adenosine receptor agonists and antagonists, has provided novel insights into the allosteric regulation of adenosine A3 (A3AR) and A1 (A1AR) receptors by allosteric ligands and receptor dimerization in single living cells [1, 2]. We have also used a fluorescent analogue of {"type":"entrez-protein","attrs":{"text":"CGP12177","term_id":"877152897","term_text":"CGP12177"}}CGP12177 to investigate ligand binding to the human β1-adrenoceptor. This work has demonstrated that there is negative cooperativity between the two different ligand-binding conformations of the β1-adrenoceptor activated by catecholamines and {"type":"entrez-protein","attrs":{"text":"CGP12177","term_id":"877152897","term_text":"CGP12177"}}CGP12177 respectively [3]. Finally, we have used fluorescence correlation spectroscopy (FCS) to investigate ligand binding to A1AR and A3AR in small 0.2 µm2 microdomains of single living cells [4]. FCS studies with a fluorescent A3-agonist have enabled high affinity labeling of the active conformation (R*) of the receptor [4]. We have also used a fluorescent adenosine A3-antagonist ({"type":"entrez-nucleotide","attrs":{"text":"CA200645","term_id":"35234116","term_text":"CA200645"}}CA200645) to study the binding characteristics of antagonist-occupied receptor conformations (R) in membrane microdomains of single cells [5]. Investigation of the dissociation kinetics of {"type":"entrez-nucleotide","attrs":{"text":"CA200645","term_id":"35234116","term_text":"CA200645"}}CA200645 provided further support for allosteric regulation of this receptor by homodimerization [5].

Perinatal complications are a serious clinical problem, in particular hypoxic-ischemic (HI) episodes, caused by birth asphyxia or uterine and fetal blood flow interruption. HI corresponds to 23% of neonatal deaths, being one of the top 20 leading causes of disease burden. Preconditioning (PC) is a stimulation below the injury threshold that activates endogenous protective mechanisms to prevent damage. Low doses of carbon monoxide (CO) play a beneficial role through PC induction. Herein, CO cytoprotection was explored in distinct brain models. The used experimental models range from monoculture of astrocytes, co-cultures of neurons and astrocytes, to the whole organism with the rat model of perinatal ischemia. In primary cultures of astrocytic cells, CO not only impairs mitochondrial membrane permeabilization, by ANT glutathionylation, but also strengths mitochondrial oxidative metabolism, by modulating COX activity, increasing mitochondrial biogenesis and ATP amounts. Also, CO reinforces astrocytes-neurons communication towards neuronal survival. Purinergic molecules are the main mediators for this non-cell autonomous effect. Our results seem to indicate that the main pathway involved includes ATP release from astrocytes, its metabolization and A2A receptor binding to initiate protective mechanisms within the neurons. Rat pups were exposed to CO before hypoxia-ischemia induction (Vannucci model). 24h after HI the brains were collected for cell death and tissue protection assessment (histological and immunohistochemical analysis). It was found limited apoptosis in hippocampus following cerebral ischemia: lower cytochrome c release and caspase-3 activation yielding an increased Bcl-2 expression. Altogether, one can conclude that there is not just a unique pathway for the CO-induced endogenous protection, brain tolerance is the result of a complex cellular change in response to injury. Indeed, CO regulates cell death pathways and modulates cellular metabolism.

MicroRNAs (miRNAs) are short, 22-25 nucleotide long transcripts that may suppress entire signaling pathways by interacting with the 3’-untranslated region (3’-UTR) of coding mRNA targets, interrupting translation and inducing degradation of these targets. The long 3’-UTRs of brain transcripts compared to other tissues predict important roles for brain miRNAs. Supporting this notion, we found that brain miRNAs co-evolved with their target transcripts, that non-coding pseudogenes with miRNA recognition elements compete with brain coding mRNAs on their miRNA interactions, and that Single Nucleotide Polymorphisms (SNPs) on such pseudogenes are enriched in mental diseases including autism and schizophrenia, but not Alzheimer’s disease (AD). Focusing on evolutionarily conserved and primate-specific miRNA controllers of cholinergic signaling (‘CholinomiRs’), we find modified CholinomiR levels in the brain and/or nucleated blood cells of patients with AD and Parkinson’s disease, with treatment-related differences in their levels and prominent impact on the cognitive and anti-inflammatory consequences of cholinergic signals. Examples include the acetylcholinesterase (AChE)-targeted evolutionarily conserved miR-132, whose levels decline drastically in the AD brain. Furthermore, we found that interruption of AChE mRNA’s interaction with the primate-specific CholinomiR-608 in carriers of a SNP in the AChE’s miR-608 binding site induces domino-like effects that reduce the levels of many other miR-608 targets. Young, healthy carriers of this SNP express 40% higher brain AChE activity than others, potentially affecting the responsiveness to AD’s anti-AChE therapeutics, and show elevated trait anxiety, inflammation and hypertension. Non-coding regions affecting miRNA-target interactions in neurodegenerative brains thus merit special attention.

Proximal Spinal Muscular Atrophy (SMA) is caused by an insufficient supply of the Survival of Motor Neurons (SMN) protein. It is characterized by the selective degradation of the alpha motor neurons in the spinal cord. Analysis of the mouse model, which faithfully mimics the SMN insufficiency and the motor neuron phenotype, shows that motor neuron degeneration starts in the axons, specifically at the neuromuscular junctions. SMN is a house-keeping factor that is necessary for the assembly of the seven-membered ring of Sm proteins around the spliceosomal snRNAs and that is therefore required for pre-mRNA splicing. Such a function cannot easily explain the selective motor neuron phenotype. It has been proposed that insufficient supply of the SMN protein affects alternative splicing, especially of U11/U12-dependent introns, in mRNAs that are crucially required in motor neurons. Details, however, remain elusive. In addition to the Sm proteins, there are the like-Sm (LSm) proteins that also form heptameric complexes and participate in various steps of mRNA metabolism. We have previously shown that one such LSm complex is involved in the transport of mRNAs to the neurites, especially the axons of motor neurons. mRNA transport and local protein synthesis is an important mechanism to bring about sudden changes in the proteome at distal regions of the cell. Here, we further explore the role of the LSm proteins in neuronal mRNA regulation and how this could be relevant for the SMA pathology. Interestingly, we find that one such LSm protein is significantly depleted in the SMA mouse model before the onset of the disease, possibly indicating a causal involvement.

The major focus of ALS research has recently moved to RNA control of motor neuron functions, as most of the newly identified genes, that alone account for more than half of ALS familial cases, are clearly associated to RNA regulation. These include FUS and TDP43, two DNA/RNA binding proteins with a role in the regulation of RNA transcription, splicing, transport and translation, and C9orf72, a gene that is marked by the presence, in carriers, of an highly expanded GGGGCC repeat that is believed to provide the mutant gene of an acquired, toxic feature by an RNA-dependent gain of function mechanism. Thus, RNA dys-metabolism is likely to represent a central issue in ALS pathogenesis. Yet, whether a specific step of RNA processing is particularly affected in ALS motor neurons is unclear. We have recently obtained evidence that a prominent effect of FUS and C9orf72 expression is the induction of stress granules-associated translational repression. In this presentation I will discuss our recent work on the molecular mechanisms underlying these effects and their potential relevance in ALS pathogenesis.

The effect of galanin is mediated through three GPCR subtypes, GalR1-3. The limited number of specific ligands to the galanin receptor subtypes has hindered the understanding of the individual effects of each receptor subtype. This review summarizes the current data of the importance of the galanin receptor subtypes and receptor subtype specific agonists and antagonists and their involvement in different biological and pathological functions.

We analyzed the pathological consequences of abnormal Wnt/β-catenin signaling in endothelial cells of brain vessels using a murine model of Cerebral Cavernous Malformation (CCM) disease that develops after endothelial-cell-selective ablation of the CCM3 gene. We report increased transcription activity of β-catenin in CCM3-knockout endothelial cells in in-vitro and in-vivo models. Such activation is cell-autonomous, independent of Wnt-receptor stimulation, does not induce canonical Wnt/β-catenin signaling and represents an early response to CCM3 ablation that initiates the expression of EndMT makers before the onset of Tgf-β/BMP signaling which is required for the progression of the pathology, as we have previously described. We also show that the NSAIDs sulindac sulfide and sulindac sulfone, which attenuate β-catenin transcription activity, significantly reduce the number and dimension of vascular lesions in the central nervous system of mice with endothelial cell CCM3 knockout. These NSAIDs thus represent pharmacological tools for inhibition of the formation of vascular lesions, particularly with a view to patients affected by the genetic variant of CCM, who continue to develop new malformations over time.

Tau is a classical microtubule-associated protein known to regulate microtubule stability in neurons. In our study, we have addressed the putative crosstalk between tau and End binding proteins 1 and 3 (EB1/3), the core microtubule plus-end tracking proteins (+TIPs), in differentiating neuronal cells. We show that tau and EB proteins interact directly and that the cellular distribution and mobility of EB proteins depends on tau localization and expression levels. Moreover, our data reveal that tau is essential for the proper localization of EB1 at the medial-distal region of the axon shaft in developing neurons. In summary, we provide evidence for a new function of tau protein as a direct regulator of EB1/3 proteins. This further indicates that the interplay between classical MAPs and core +TIPs may be important for the fine-tuned regulation of microtubule dynamics and stability during neuronal differentiation.

Ginkgo biloba extract is a neuroactive agent that is widely used for correction of age-associated impairment of memory, attention deficit and other cognitive functions. It has been shown that ginkgolides and bilobalides, Ginkgo biloba extract components, are potent blockers of glycine receptors (Kondratskaya et al., 2002, Hawthorne et al., 2006). However, the effect of ginkgolic acid, the other Ginkgo biloba extract constituent, on ligand-gated ion channels was not investigated. In the present study, using patch-clamp technique and transient transfection of different subunits in CHO cells we have shown that glycine receptors (GlyRs) are modulated by ginkgolic acid in a subunit-specific manner. After pre-application of ginkgolic acid (0.5-2 min), glycine-induced currents mediated by α1 GlyRs were strongly potentiated, while currents mediated by α2 GlyRs exhibited weak inhibition. There was no significant effect of ginkgolic acid on amplitudes of currents mediated by α3 GlyRs or on GABAARs composed of 1α/1β/2γ subunits. In order to further investigate subunit-specific effect of ginkgolic acid we have focused on possible interaction sites for this compound inside different GlyR domains. We found that mutation of three residues (T59A/A261G/A303S) in α2 subunit can convert the inhibitory action of ginkgolic acid into potentiation. Indeed, application of ginkgolic acid to cells expressing α2 T59A/A261G/A303S subunits resulted in an increase of responses to low concentrations of glycine and abolishment of the inhibitory effect, typical for wild type α2 GlyR. Our results suggest that (i) ginkgolic acid selectively enhances the function of α1 GlyRs and attenuates the function of α2 GlyRs, (ii) mutation of α2 subunit converts effect of ginkgolic acid from inhibition to potentiation.

Regulated trafficking of mitochondria in neurons is essential for providing ATP at the correct spatial location to power neural function and computation, and for providing calcium buffering at sites of calcium entry or release. Indeed the regulation of mitochondrial distribution, morphology and function are proposed to play an important role in neuronal development and survival but the regulatory mechanisms remain unclear. Miro family proteins (Miro1 and Miro2 in mammals) contain a transmembrane domain locating them to the outer mitochondrial membrane, along with two GTPase domains and two calcium-sensing EF-hand domains that face into the cytosol, and play a key role in regulating mitochondrial transport. Miro proteins mediate mitochondrial trafficking in neurons by linking mitochondria to kinesin and dynein motor proteins for their transport in axons and dendrites. Miro proteins are also targets for the Parkinson’s Disease associated PINK1/Parkin mitophagy pathway and are therefore implicated in altered mitochondrial dynamics during mitophagy. Here I will present our recent results on the role played by Miro proteins in regulating mitochondrial trafficking and quality control. The role that Miro-mediated control of mitochondrial trafficking and turnover plays in regulating neuronal development, function and pathology will also be explored.

Brain tumors represent a recognized cause of epilepsy in both children and adults. In principle, any tumor (extra-axial, intra-axial, benign or malignant, common or uncommon) can cause seizures. However, patients with supratentorial low-grade glial tumors are more likely to develop epilepsy. Several clinical studies emphasize that pharmacologically intractable epilepsy critically affects the daily life of patients with brain tumors, even if the tumor is under control. Recently, the term of long-term epilepsy associated tumour (LEAT) has been introduced. LEATs are low grade, slowly growing, cortically-based tumours which predominantly occur in young patients with long histories (often 2 years or more) of drug-resistant epilepsy. Glioneuronal tumors (GNT), including gangliogliomas (GG) and dysembryoplastic neuroepithelial tumors (DNTs), represent the most common tumor within the spectrum of LEAT. The advent of the neurosurgical treatment of epilepsy-associated brain lesions confirmed the strong epileptogenicity of these tumor entities. Understanding the mechanisms that underlie epileptogenesis in LEATs is essential to identify new treatment targets and to develop an effective therapy. Mechanisms such as alterations of the balance between excitation and inhibition and alterations in neuron-glia interactions might be involved. Astroglial cells express functional receptors for a variety of neurotransmitters and may critically modulate synaptic transmission. In addition, an increasing number of observations indicate that pro-epileptogenic inflammatory pathways are activated in GNT and may contribute to the onset and progression of seizures. The recent advances and likely candidate mechanisms and molecules involved in tumor-associated epileptogenesis will be discussed.

Post-translational modification of the neural cell adhesion molecule (NCAM) by polysialic acid (PSA) is crucial for nervous system development and brain plasticity. PSA attachment is catalyzed by the two polysialyltransferases, ST8SiaII and ST8SiaIV. ST8SiaII dominates during embryonic and early postnatal development while ST8SiaIV is the prevailing enzyme of the juvenile and adult brain. A growing body of evidence links aberrant levels of NCAM and PSA to neuropsychiatric disorders, including schizophrenia. To investigate whether polysialyltransferase deficiency might cause a schizophrenia-like phenotype, ST8SiaII-/- mice, ST8SiaIV-/- mice and their wild-type littermates were assessed neuroanatomically and subjected to tests of cognition and sensory functions. ST8SiaII-/- but not ST8SiaIV-/- mice displayed enlarged lateral ventricles and a size reduction of the thalamus accompanied by a smaller internal capsule and a highly disorganized pattern of thalamocortical and corticothalamic fibers. Loss of ST8SiaII was associated with reduced phosphorylation of fibroblast growth factor receptor 1 in the frontal cortex of newborn mice and retarded neuronal differentiation of newly generated cells in the dentate gyrus of adults. ST8SiaII-/- and ST8SiaIV-/- mice were both impaired in short- and long-term recognition memory, but only ST8SiaII-/-mice displayed impaired working memory and a deficit in prepulse inhibition, which could be attenuated by clozapine treatment. Furthermore, ST8SiaII-/- mice exhibited anhedonic behavior and increased sensitivity to amphetamine-induced hyperlocomotion. These data reveal that reduced polysialylation in ST8SiaII-/- mice leads to pathological brain development and schizophrenia-like behavior. We therefore propose that ST8SiaII deficiency has the potential to cause a neurodevelopmental predisposition to schizophrenia.

Orexins A and B (hypocretins 1 and 2) are two closely related peptides produced mainly by hypothalamic neurons that project to numerous brain structures. Orexins exert their biological activity by binding to two subtypes of GPCR receptors, OX1R and OX2R. The orexin system has been shown to orchestrate multifaceted physiological functions, including vigilance and the sleep/wake cycle, energy homeostasis, endocrine, visceral functions and pathological states, such as narcolepsy and drug abuse [1]. In addition, a neuroprotective potential of orexin A has been recently demonstrated in rats using a model of focal cerebral ischemia [2]. In our studies we investigated effects of orexins on survival of cultured neurons from the rat cerebral cortex. Quantitative real-time PCR revealed the presence of OX1R and OX2R in cortical neurons. Orexins and [Ala11-D-Leu15]orexin B (a selective agonist of OX2R), used at 0.001-1 microM concentrations, markedly increased neuronal cells viability, an effect associated with an attenuation of caspase-3 activity. Comparable potency of the three peptides suggests a predominant role of OX2R in the studied phenomenon. Under conditions of chemical hypoxia, orexins potently increased neuronal viability and protected cortical neurons from oxidative stress. The pro-survival effect of orexins was blocked by U0126 and 10-DEBC, inhibitors of MEKK and Akt, respectively. In addition, orexins A and B stimulated Akt and ERK1/2 in cortical neurons in a time- and concentration-dependent manner. It is suggested that both Akt and ERK1/2 play an important role in the pro-survival effects of orexins in neurons.

Peripheral nerve injuries are an important part of everyday medical practice since there are reported over 600,000 cases in Europe and in the United States annually [1]. Many of such nerve injures cause gaps between nerve stumps. Without intervention, they can lead to the formation of a stump neuroma, what can result in functional impairment of the nerve fiber. The current approaches to regeneration of damaged peripheral nerves include: autografting, allografting, and, last but not least, the implantation of polymeric tubes and conduits between nerve stumps. Nerve autografting is considered as the “gold standard” technique for the repair of peripheral nerve discontinuities, but it has a number of limitations, such as the requirement for the second surgical procedure to harvest the graft tissue, the donor site morbidity, additional injuries and scarring as well as the increased recovery time. Allografts (e.g., cadaver nerve grafts) and xenografts (e.g. animal nerve grafts) can be an alternative to autografts, but their main drawback lies in the high possibility of an undesirable immune response. The most promising materials for peripheral nerve conduits preparation are natural biopolymers. They are obtained from natural sources, exhibit similar properties to the tissues they are replacing and reveal good cell adhesion. Furthermore, they tend to accelerate regeneration processes due to specific chemical interactions within the human body, e.g. with extracellular matrix (ECM) molecules. The purpose of our study is to create a conduit with properties that will mimic the ones of the extracellular matrix of the peripheral nervous system. Our focus is put on natural polymers, especially chitosan. In addition, we use chemotactic factors which exhibit properties beneficial in regeneration of the peripheral nervous tissue.

The nucleus accumbens has long been a major target for studies on the rewarding effects of drugs of abuse or physiological reinforcers, whereas the brain regions medial of the medial accumbens shell have received less attention. We investigated if counterconditioning with dyadic (i.e., one-to-one) social interaction, a strong inhibitor of the subsequent reacquisition of cocaine conditioned place preference (CPP), differentially modulates the activity of the diverse brain regions oriented along a mediolateral corridor reaching from the interhemispheric sulcus to the anterior commissure, i.e., the nucleus of the vertical limb of the diagonal band, the medial septal nucleus, the major island of Calleja, the intermediate part of the lateral septal nucleus, and the medial accumbens shell and core. EGR1 activation was predominantly found in dynorphin-labeled cells, i.e., presumably D1 receptor-expressing medium spiny neurons (D1-MSNs), with D2-MSNs (immunolabeled with an anti-DRD2 antibody) being less affected. Cholinergic interneurons or GABAergic interneurons positive for parvalbumin, neuropeptide Y or calretinin were not involved in these CPP-related EGR1 changes. Glial cells did not show any EGR1 expression either. Cocaine conditioning increased the spike frequency of neurons in the septal nuclei, whereas social interaction conditioning increased the spike frequency in the nucleus accumbens compared to saline control animals. In addition, social interaction conditioning decreased the amount of active neuron clusters in the nucleus accumbens. The present findings could be of relevance for the therapy of impaired social interaction in substance use disorders, depression, psychosis, and autism spectrum disorders.

Autism spectrum disorders (ASD) are heterogeneous, heritable neurodevelopmental conditions, affecting ~0.5% of the population across cultures, with a ~4:1 male/female ratio. ASD are characterized by social interaction and communication deficits, restricted interests, repetitive behaviors, and reduced cognitive flexibility. Causes likely converge at the synapse, as shown by mutations of synaptic genes or mutations causing quantitative alterations in synaptic protein expression. Neuroligin4 (NLGN4X) mutations are among the most frequent causes of heritable ASD. But monogenetic forms altogether account for 1200 schizophrenic subjects and validated it in Asperger autists. We hypothesized that a coincidence of unfortunate normal variation in synaptic or synapse-regulating genes rather than mutations underlies most autistic phenotypes. We identified ‘proautistic’ variants in synaptic genes, which in aggregate are associated with high autism severity. A transcranial magnetic stimulation study on respective individuals revealed enhanced glutamatergic and GABAergic activity. IPS-derived cortical neurons from these subjects are now functionally characterized.

Diabetic retinopathy (DR) is a neovascular, inflammatory and neurodegenerative disease. The neovascular component is treatable but no therapeutic agents are available for the other two components. Early changes in the diabetic retina include neuronal death of amacrine and retinal ganglion cells (RGC)(Barber et al., J Clin Invest,1998), and elevated levels of inflammatory mediators (Yoshimura et al., Plos One,2009). Nerve Growth factor (NGF) receptors, namely TrkA and p75NTR, are expressed in RGC. The TrkA receptor activates prosurvival, while p75NTR activates inflammatory and apoptotic pathways (Mysona et al., Expert Rev Ophthalmol,2014). Dehydroepiandrosterone (DHEA) binds to both receptors, mimicking NGF. It affords anti-apoptotic, neuroprotective and anti-inflammatory effects in the retina (Kokona et al., Neuro-pharmacol, 2012, Straub et al., J Clin Endocrinol, 1998). The therapeutic use of DHEA is restricted due to its metabolic products. The main objective of this study was to investigate and compare the neuroprotective and anti-inflammatory effects of DHEA and its spiro-epoxy derivatives BNN27 and BNN20(Calogeropoulou et al., J Med Chem,2009) (not metabolized to estrogens and androgens), in the rat streptozotocin model of DR. BNN27, via TrkA activation, protected in a dose-dependent manner (2, 10, 50mg/kg, ip) bNOS (brain nitric oxide synthetase) and TH (tyrosine hydroxylase) expressing amacrine cells and ganglion axons (NFL immunoreactivity) similar to DHEA’s actions, while BNN20 was less effective. BNN27 activated the TrkA prosurvival signaling pathway ERK1/2 kinase. It reduced the activation of SAPK/JNK kinase and the expression of p75NTR. BNN27 also increased the expression of anti-inflammatory cytokines (IL10). These results suggest that NGF TrkA receptor is involved in the neuroprotective and anti-inflammatory effects of BNN27 and is a valuable target via which BNN27 could afford efficacious therapeutics for the treatment of DR.

During cerebellar development, granule cell precursors are produced from a secondary germinative zone forming the external granule cell layer (EGL). Immature granule neurons from the inner part of the EGL then start a tangential migration followed by a centripetal inward radial migration across the molecular and Purkinje cell layers to reach their final destination at the bottom of the forming internal granule cell layer (IGL). This complex migratory process is highly regulated and takes about 2 days in rodents and it is essential for the proper formation of the cortical layers forming the mature cerebellum. In the IGL, granule cells differentiate to establish functional excitatory synapses with GABAergic neurons including Purkinje, basket, stellate and Golgi cells, or die. Some neurotrophins and neurotransmitters have been shown to be involved in the control of cerebellar granule cell survival, migration and differentiation. Initially, when I started my carrier as a researcher, we used to claim that very few neuropeptides were produced in the cerebellum. Nevertheless, we now know that this was wrong as we have recently identified by mass spectrometry over 70 peptides expressed in the cerebellum during development. Over the years, the involvement of some of these peptides such as somatostatin, PACAP or ODN, has been established in the control of cerebellar granule cell survival, migration and differentiation as will illustrate my presentation.

Spinal and bulbar muscular atrophy (SBMA) and amyotrophic lateral sclerosis (ALS) are motoneuron diseases. A mutation in the androgen receptor (ARpolyQ) gene is responsible for SBMA. Mutations in the SOD1, in the TDP-43, in the FUS-TLS or in the C9ORF72 genes are responsible for familiar form of ALS. The mutated coded proteins misfold and aggregates. Efficient protein quality control (PQC) is required for the maintenance of physiological and soluble protein pool in affected motoneuron. The balance between autophagy and ubiquitin-proteasome system (UPS) prevents protein aggregation and increases degradation of SBMA and ALS misfolded proteins. Dynein binds the co-chaperone BAG3 and transports the mutant proteins at microtubule organization center where misfolded proteins interact, aggregate and can be degraded by autophagy. However, here misfolded proteins may blocks autophagy flux. In NSC34 cells, dynein is sequestered into ARpolyQ aggregates suggesting the role of dynein into aggregates formation process. Unexpectedly, the silencing of dynein heavy chain resulted in a drastic reduction of ARpolyQ retained in filter retardation assay (FRA). Moreover, dynein silencing drastically altered autophagic markers localization (LC3 and p62) by immunofluorescence. Notably, treatment with a dynein inhibitor (EHNA) drastically reduced the retention of ARpolyQ, mutSOD1 and mutTDP43 aggregates in FRA, even when autophagy was inhibit with 3-MA. Conversely UPS blockage with MG132 counteracted the reduction induced by altered dynein transports. RTq-PCR on NSC34 cells treated with EHNA showed an increased BAG1:BAG3 ratio that can targeting the misfolded proteins to UPS. Moreover, in NSC34 cells, EHNA increased the degradation of proteasome reporter GFPu, while BAG1 overexpression reduced the level of aggregates retained in FTA. These data suggest that, when autophagy is overload, by misfolded proteins, dynein inhibition restores the physiological and soluble protein pool via UPS.

A number of different genes have been found mutated in patients with Amyotrophic Lateral Sclerosis (ALS). Several of these genes encode for proteins involved in multiple steps of RNA processing, suggesting that mRNA dys-metabolism has a role in the degeneration of motor neurons. This is the case also for FUS-linked ALS. FUS (Fused in Sarcoma) is a DNA/RNA binding proteins with an established, yet not completely clear, role in the regulation of RNA transcription, splicing, transport and translation. However, recent evidence indicates that (similarly to mutant ALS-linked SOD1) the toxic function of this protein may lie also in its propensity to aggregate and sequester other proteins, and/or in its ability to induce mitochondrial damage and oxidative stress. In this presentation I will discuss our recent work on the molecular mechanisms underlying these effects and their potential relevance in the pathogenesis of ALS.

G protein–coupled receptors (GPCRs) are intensely studied as drug targets and for their role in signaling. High-resolution crystal structures of GPCRs capturing different receptor conformations are now available, which have provided insights into the mechanism of activation and ligand selectivity for this important class of drug targets. I will first present a series of structure-based screens for novel ligands of the A2A adenosine receptor (A2AAR), which is a drug target for Parkinson’s disease (antagonists) and ischemia (agonists). As crystal structures for both inactive- and active-like receptor conformations of the A2AAR have now been determined, molecular docking screens for novel ligands can be performed. Virtual screens against different conformations of the A2AAR were carried out to investigate if structure-based methods can be used to identify agonists and antagonists. Our results shed light on the importance of access to crystal structures and the role of the chemical library in screens for ligands with specific pharmacological properties. For most GPCRs, no experimental coordinates are available and structure-based screens are forced to rely on homology models. However, it is still unclear if models of GPCRs are sufficiently accurate to be used in ligand discovery. The determination of crystal structures for dopamine and serotonin receptors, and the challenges to the community to predict these in the GPCR Dock competitions, have enabled us to carry out comparisons of ligand discovery from models versus crystal structures. Our results from these challenges reveal opportunities and limitations of the use of homology models in ligand discovery and design of selective lead candidates.

TDP43 is a ubiquitously expressed prevalently nuclear protein involved in RNA splicing, RNA stability and miRNA processing. Amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD) and inclusion body myopathy (IBM) are characterized by TDP43 being depleted from the nucleus and accumulating in cytoplasmic inclusions, which are the pathological hallmark of the disease – these diseases are also defined as “TDP43 proteinopathies”. Mutations in TDP43 have been found to be causative of a proportion of ALS cases reinforcing the primary importance of this molecule in disease pathogenesis. The pathogenic mechanism by which TDP43 acts is unclear, and both loss of nuclear function (LOF) and gain of function (GOF) mechanisms have been proposed. We here discuss how we used muscle tissue, which provides high quality RNA of patient disease tissue, to investigate the consequences of TDP43 mislocalization. Further, we characterize two novel mouse TDP43 mutant lines, carrying ENU-induced mutations in order to study the effects of TDP43 mutations expressed at physiological levels in the mammalian central nervous system. One mutation (deltaRNA) strongly reduces the RNA-binding capacity of the protein; the second mutation (C-TERM) is in the glycine-rich C-terminal domain where the majority of human pathogenic mutations are found. Our results underline the importance of studying models with physiological expression levels of TDP43 mutations and shed light on the different effects on RNA metabolism caused by the TDP43 loss and gain of function.

Neurogenic inflammatory components mediated by peptidergic sensory nerves have a crucial impact on the symptoms of inflammatory diseases. Galanin is a regulatory sensory neuropeptide, which was shown to attenuate neurogenic inflammation, but our current understanding about its endogenous targets, and physiologic importance is incomplete. Among the endogenous receptors of galanin (GAL1, GAL2, GAL3) we found GAL3 to be the most abundantly expressed on the vasculature and GAL2 on different types of immune cells including polymorphonuclear neutrophils and natural killer cells. Therefore, we evaluated if galanin exerts direct or indirect effects on these immune cells. Our data revealed that galanin can be regarded as an immunomodulatory peptide as it can sensitize polymorphonuclear neutrophils and natural killer cells towards proinflammatory cytokines. Since there are only scarce in vivo data concerning the role of GAL3 in inflammatory disease conditions, we analysed its involvement in the K/BxN serum transfer model of autoimmune arthritis and the oxazolone-model of allergic contact dermatitis, employing GAL3 gene-deficient mice. After arthritis induction, GAL3-knockout mice demonstrated increased clinical disease severity and earlier hindlimb edema than wildtype mice. Vascular hyperpermeability was also elevated compared to wildytpes, but neutrophil myeloperoxidase activity and arthritic hyperalgesia were not significantly different. In contrast, disease severity, vascular, and immune components were not affected in allergic contact dermatitis in GAL3 knockouts in comparison with wildtypes. Our findings suggest GAL3 activation as a substantial anti-inflammatory pathway in neutrophil-dominated autoimmune arthritis, modulating the early neurogenic vascular hyperpermeability and consequent edema formation. However, the involvement of GAL3 activation in the T-cell dependent allergic contact dermatitis remains unsupported.

Stopping the succession of events that lead to development of Parkinson’s disease (PD) is a principal challenge that requires a brain protective approach in early diagnosed patients. Although PD ethiopathology may not have a single causative factor, information from sporadic and familial cases together with chemical and genetic animal models strongly suggests that low-grade chronic inflammation and oxidative stress play a critical role. Our team is studying the relevance of the transcription factor NRF2 (Nuclear factor (erythroid-derived 2)-like 2), a master regulator of oxidant and inflammatory defense, as a new therapeutic target in PD. The pro-inflammatory activation of microglia and astroglia in response to LPS, MPTP and human α-synuclein over-expression is exacerbated in Nrf2-deficient mice, thus demonstrating an immunomodulatory role of this protein. In PD patients, some evidence gathered from epidemiological, genetic and anatomopathologic studies also support a protective role of NRF2. Several compounds activate NRF2 and provide an immunomodulator and cytoprotective response in preclinical animal models of PD. At this time a crucial point to translate these promising results to the clinic is the discovery of a drug with good pharmacokinetics and pharmacodynamics that fulfill criteria of safety, tolerability and efficacy. The use of repurposing drugs, such as dimethyl fumarate used nowadays for treatment of remitting relapsing multiple sclerosis, may provide excellent candidates.

Oligodendrocytes, the myelin forming cells of the CNS, enwrap neuronal axons and form multilamellar myelin sheets. They are derived from oligodendrocyte precursor cells which migrate from the subventricular zone into the different regions of the brain. Differentiation from the early progenitor to the mature multiprocessed oligodendrocyte is characterized by different morphological stages. To support cell morphology and establish and maintain the myelin membrane, an intact, spatially organized cytoskeleton with dynamic properties is essential. In particular microtubules and their associated proteins play an important role. A variety of microtubule binding proteins, including tau, are present in oligodendrocytes. Oligodendrocytes in culture express all six isoforms of tau which are developmentally regulated. Tau proteins are present in immature and mature oligodendrocytes and specifically prominent in the branching points of the cellular processes. Downregulation of tau impairs cell differentiation and the process of early myelination. In neurodegenerative diseases collectively termed tauopathies, fibrillary tau accumulations occur not only in neurons but also in glia. Tau positive coiled bodies originating in oligodendrocytes are characteristic for the brains of patients with frontotemporal dementias, such as corticobasal degeneration and progressive supranuclear palsy. These aggregates are further characterized by the presence of heat shock proteins and ubiquitin, indicating that stress situations are causally related to the pathogenesis. In this respect, proteasomal dysfunctions have been discussed to be involved in neurodegenerative disorders and the aging process. Data on the consequences of proteolytic stress in oligodendrocytes and the protein aggregation process will be presented. Our data demonstrate that an intact cytoskeleton is essential for cellular defense mechanisms.

Background Alzheimer’s Disease (AD) Research has long been focusing on Ab-containing amyloid plaque deposition and tau-containing tangles. However, recent clinical trial outcomes by Ab-lowering approaches have been disappointing. As an alternative approach, the present study focuses on mechanisms that prevent neuronal loss, another pathological hallmark of AD. Specifically, we have uncovered an unexpected role of glial cell line-derived neurotrophic factor (GDNF) in AD.

Infectious diseases exert a major burden of disease in developing countries with 99% of the global burden of infectious diseases, as measured by DALYs, in low and middle income countries. While better use of existing vaccines would make an appreciable difference, the greatest burden is caused by diseases for which we currently have no vaccines. The picture, especially in children, is dominated by diarrheal and respiratory diseases. Paradoxically these diseases have relatively low priority for funding in absolute terms, and especially in relationship to the burden of disease. Thus, new vaccines for these neglected diseases need both innovative scientific solutions and innovative development schemes involving scientific institutes, public financing and industrial input. The industrial input is critical: not only will vaccine manufacture require an industrial partner, but the knowledge to efficiently undertake the technical and clinical development leading to vaccine production largely resides in industry. A potentially important development in this area has been the recent formation of Industry Linked Vaccine Institutes: For example, the Novartis Vaccines Institute for Global Health and the Hilleman Laboratories. These are an important conduit for applying industrial know how for developing commercial vaccines to the pressing need for vaccines for neglected diseases of developing countries.

The discovery of HIV in 1983 originated from a collective adventure, which mobilized clinicians, researchers and patients altogether. This collaboration was crucial to rapidly expand the knowledge of the virus and develop the first diagnostic tests and antiretroviral therapy (ART). More than 25 years after the discovery of the etiological agent responsible for AIDS, research priorities still remain care, treatment and prevention with the major objective of developing a preventive vaccine. Today, we have gained significant insight into the virus pathogenesis. The evolution and progression of the disease caused by HIV is closely linked to a number of determinants of the virus itself and the host. We also know today that, very early after exposure to the virus, a massive depletion of CCR5+ CD4+ T memory cells associated with microbial translocation occurs in the gastrointestinal tract of HIV infected patients. HIV infection is clearly inducing an inflammation and a generalized and persistent T cell activation, which may play a role in the persistence of HIV infection, resulting from the establishment of permanent reservoirs into host cells and in different host compartments. The reduction of the size of these reservoirs is representing one of the main challenges for the development of future therapeutic strategies. Among other challenges in therapy, we also need to better understand the mechanisms leading to the severe complications (cardiovascular diseases, accelerated aging, cancer...) observed in some patients on long-term ART. Again, whether the inflammatory response is contributing or not to these complications remains an opened question. The early acute phase of HIV infection appears therefore to be crucial in determining disease progression. Given the importance of the innate immune responses in this very early phase following infection and in driving adaptive immunity, further research on innate immunity in HIV infection are certainly among priorities for elaborating future therapeutic and vaccine strategies. New technologies are today available to address all these scientific challenges. But they will only be overcome with a multidisciplinary and translational research for the global benefit of humanity.

During the last decades, whole chains of dengue epidemics emerged in the South Pacific region. In contrast with the situation in hyper endemic continental countries and in the Caribbean, the epidemiology of dengue in the South Pacific islands Countries (SPICs) is characterized by the non-persistent co-circulation of multiple serotypes and the long-term predominance, with local re-emergences, of a single genotype. Local specificities in the epidemiological profile of dengue can also been observed between the SPICs, probably related to differences in the geographical situation, the eco-biological context (climate, endemic mosquito species), the demography and the population flew. In the present study, by focusing on the past DEN31989-96 and the recent DEN12001-10 and DEN42007-10 circulation periods, we addressed the question of the circulation of dengue viruses between the French Pacific Territories and the impact of the “local context” on viral genetic evolution. Hundreds of viral strains collected during both epidemic and endemic periods in French Polynesia (FP), New Caledonia (NC) and Wallis & Futuna (WF) were sequenced on the complete E gene. The phylogenetic analysis corroborates the previous observations on the predominant circulation of a single genotype. Within each serotype/genotype, the viral strains collected in the SPICs formed a “Pacific clade”. Within this clade, the strains from the French Pacific Territories formed a unique lineage during the early epidemic/endemic circulation periods but diverged in distinct lineages when the virus re-emerged. By analyzing the in time/in space fixations of genetic mutations on the E gene, we observed that some mutations are shared by the French Pacific Territories but differ from the other SPICs. Moreover, although the majority of the mutations acquired in FP are also found on NC and WF strains, some seem to be specific to the Territory. Our results support the hypothesis of an impact of both the regional and local contexts on the genetic evolution of dengue viruses.

Nowhere in the world are the health impacts from emerging diseases of wildlife more important than in developing countries, where daily work and livelihoods are highly dependent on natural resources. Many of these same countries have little to no capacity for detecting disease emergence in wildlife and domestic animals prior to spread to humans. While the linkages of human, animal, and environmental health is at the heart of the One Health approach, an increasingly important prism through which governments, NGOs, and practitioners view public health, we still have three critically important challenges facing us: 1) we need a much broader and deeper knowledge of what pathogens are waiting to emerge from the animal kingdom, 2) we need to better target our investigations to maximize available resources, and 3) we need better tools to predict or determine if an organism is a pathogen of significance for causing human disease. The PREDICT project of the USAID Emerging Pandemic Threats program is endeavoring to build capacity for surveillance of emerging diseases in wildlife in order to help to address these three challenges.

Based on the knowledge that a cured infection protects the individual from re-infection, the development of a vaccine to prevent leishmaniasis has been a goal for nearly a century. Indeed, it is generally believed that after healing of leishmaniasis, sterile cure is never achieved and that few residual living parasites will remain sequestered within some host cells that offer them a safe shelter and hence maintain anti-parasite immune memory. This statement is mainly supported by data from experimental leishmaniasis in mice of susceptible or resistant phenotype, in which, live parasites could be recovered from lesions even after healing, and in which disease reactivation can be obtained by immune manipulation even after apparent complete cure. Whether maintenance of a long-term immune effector memory in humans will also require persistence of live parasites is presently unknown but is very important in the perspective of a vaccine development. Our aim herein was to address the issue of Leishmania major parasite persistence vs. sterile healing in zoonotic cutaneous leishmaniasis (ZCL) by analyzing biopsies of scars from healed volunteers. Skin-punch scars’ biopsies (n=59, range of scar age: 1– 5 years) have been obtained from volunteers (18–55 years old) living in two ZCL endemic foci, who had a confirmed past history, are clinically cured of ZCL and who gave their written consent. The whole protocol was approved by the local IRB. The specimens were taken under sterile conditions and local anaesthesia. Each specimen was divided into three parts: (i) the first sample was processed for quantitative real time PCR, (ii) the second was cultured in vitro in enriched medium and (iii) the third was inoculated into the footpad of susceptible BALB/c mice which were kept under observation for five months. For in vitro isolation and after microscopic observation for at least 8 weeks, all cultures were found negative. For in vivo isolation, biopsy-inoculated mice were killed five months later, and skin fragments, draining lymph nodes and spleen were inoculated into culture medium, observed carefully for at least 8 weeks before being also designated as negative. PCR results were found negative for the majority of biopsies. These results indicate that any potential persistent living parasites after cure are unlikely to be sequestered in ZCL scars. This issue is of importance in strategies for control of leishmaniasis and requires further discussion.

There are four dengue virus serotypes, based on molecular analysis. Each serotype is divided to several different genotypes, such as there are 5 genotypes in dengue virus serotype 3 (DENV-3), in which genotype II was confirmed to be the main etiology cause the big dengue fever/dengue hemorrhagic fever outbreak in Vietnam 1998. Molecular epidemiological study was carried out on DENV-3, which causing dengue fever/dengue hemorrhagic fever in some epidemic areas with high morbidity and mortality from southern Vietnam in 2010. In this study, viral RNA of DENV-3 were extracted directly from dengue fever/dengue hemorrhagic fever patient sera, typing by RT-PCR (reverse transcriptase-polymerase chain reaction) with Lanciotti specific primers, sequencing the entire envelope (E) gene and comparative analysis with DENV-3 strain isolated from the big outbreak of dengue hemorrhagic fever in whole country in 1998. The phylogenetic analysis of DENV-3 from Vietnam as well as previously published strains, the results showed that: Vietnamese DENV-3 belong to genotype II among 5 genotypes classified for worldwide DENV-3 and have been closely related to Thailand isolates. Further researchs need to be carried out in order to confirm the evidence for local genetic evolution and conserved functional structure of E protein of DENV-3.

The role of treatment of vascular lesions of the legs is very important in the elderly. Pain, inability, bandages, periodicity of medications can seriously interfere with a good quality of life because of the lower compliance of geriatric patients to accept a surgical approach. To allow a new approach to vascular lesions the Authors propose a guideline in which the topic of “less invasive” surgery is underlined based on their pathophysiological causes. Patients affected with lesions of the legs are addressed to the Leg Ulcers Centre by general physicians and selected on the basis of age and comorbility to propose a therapeutical plan. General conditions, evaluation of lesions healing and of quality of life are correlated to final target (acceptable quality of life). The first step includes a surgical examination and cardiological, nephrological, neurological examinations. The following steps are different for pathologies (see Flow Chart). PAD (Peripheral Arterial Disease) The target is the limb revascularisation. It can be allowed by endovascular techniques. Vascular evaluation is obtained by a ultrasonographic study followed by an Angio-Tc study of the arterial district. In diabetic patients endovascular techniques are used to obtain therapeutical windows in which skin closure occurs. Prostacyclin and prostanoids are very useful in those patients in which a surgical procedure is not possible. They can be administred by e.v. at low doses for 2-3 days using a reservoir. Open surgery is restricted to revascularization of profunda femoral artery. Venous insufficiency The target is to decrease venous hypertension in the lesion district. Surgery is useful to heal the lesion, but also to prevent recurrency and to decrease elastic compression. For a surgical approach an haemodynamic map is necessary to propose operations (see Figure ​Figure11) Figure 1 Topical treatment of the ulcer Surgical debridment of the ulcer can be obtained under local anesthesia using anesthetic ointment. According to the patient’s compliance homologous skin grafts or dermal substitutes can be used with good results. Amputations Minor amputations allow antidolorific drugs administration to be decreased. The final result can be accelerated by reconstructive plastic surgical procedures or by implant of homologous skin grafts. All patients have accepted surgical treatment and when necessary another option. The Authors suggest the role of the internist and of the anesthesist in the management of different comorbidiities and the use of less invasive techniques, underlining that this is a proposal to obtain a guideline for the therapy of skin lesions in the elderly.

Bladder incontinence is present in all age groups. But risk factors such as increasing age, degenerative diseases, intervention due to prostatic diseases, metabolic or neurological disorders which are typical of an advanced age result in a challenging reality to face. It's a widespread custom to consider incontinence as a disorder or a natural consequence of aging; on the contrary, it is a disease that responds well to various treatments. Urinary incontinence of the elderly is a complex issue because it determines significant changes in personal lifestyle causing serious psychological states of depression. Losing control of our body basic functions can lead busy people to lifestyles never before considered, often self-limiting. The item requires an integrated approach to the various professions in order to allow people the best life quality. Good advice and information can help to handle and resolve one of the main causes of hospitalization or long stays in healthcare facilities.

Lymph-oedema is a chronic disease, not easily controllable with evolutive, progressive tendency. Since 2001 in the angiological outpatients’surgery inside of Operative Unit of Territorial Rehabilitation, in the ambit of patients affected by lymph-oedema, also the rehabilitative projects of geriatric patients affected by secondary post-surgery lower and upper limb lymph-oedema, contained the therapheutical programs, have been opening by angiologist and physiotherapist, in charge of the patient, together: in the older patients oncological lymph-oedema is more common than benign lymph-oedema tardum and furthermore, in the elderly most accompanying disease aggravate this pathology. Lymph-oedema requires careful patient monitoring that consists of: 1.the first evaluation performed in the ambit of a multidisciplinary and multi-professional team that leads to the compilation of a specific clinical report and a rehabilitative project which personalized therapeutical programs, adequate to the treatment of disabilities according to the evolution stage. The clinical report, point of reference in the next check-up, includes: - anagraphical data, anamnesis, clinical examination, evaluation of articulation limitations and eventual functional impotencies, volumetrical evaluation; - instrumental examination: since 2005 we have been using Echo-colour-Doppler diagnostic useful for tissue examination because it provides important indications on structural characteristics of examined tissue; while the angiologist is carrying out the exam, the physiotherapist fills up a sheet contained a limb drawing useful for a more personalized therapeutical program; - individual rehabilitation training project; 2.carefull lymphological monitoring of patients to assess rehabilitative training effectiveness 3.patients’ follow-up: periodical medical check-up who performed physical treatment, to plan an eventual rehabilitative training based on clinical-instrumental case history.

Background The decreased mortality rate obtained with antiretrovirals (ARVs) induced a shift of HIV infection toward a chronic condition and increased the fraction of patients aged >50years. Senior patients can be included in three subgroups with different clinical problems. Long term exposure to ARVs give rise to the main clinical problems. Several studies recognize age as an independent risk factor for acute and chronic toxicities probably because of an additive mechanism with age related health problems (metabolic syndrome, cardiovascular diseases, endocrine-metabolic, liver, renal, bone and central or peripheral nervous system impairment. Among 1220 HIV we observed a higher rate of switching/discontinuation due to toxic effects in the elderly (Figure ​(Figure1).Interaction1).Interaction between ARVs and several drugs widely used in the elderly (H2-Antagonists, Pump-Proton-Inhibitors, Benzodiazepines, Calcium-Channel-Blockers, Hormonal-Contraceptives, HMG-CoA-Reductase-Inhibitors, Phosphodiesterase-Type-5-Inhibitors, and Oral-Anticoagulants) are reported to and can influence both treatment strategy and effectiveness in older patients. Figure 1 Comparison of the causes for ARVs switching or discontinuation in senior and younger HIV infected patients. Causes of switching/discontinuation of antiretrovirals regimen in 1220 HIV positive patients on HAART followed up in the II Division of Infectious ... Late primary infection Acute infections are hardly recognized but, since most HIV infections acquired in advanced ages are sexually transmitted, association with other STI is expected in this subset of patients who are not aware of own their risk, mode of HIV transmission and seem poorly reached by health education programs on safe sexual behaviour. Post-menopausal women are at a higher risk of infection due to hormonal mucosal changes and decreased use of condoms.

In our country too, the growing number of elderly people together with the contemporary lowering of the birth rate, has now, and will continue to have in the near future, strong influence on choices to be made in healthcare and on welfare policy as a whole. Therefore, the healthcare system must redefine its priorities and face a series of questions regarding the elderly. It will be necessary to rethink care giving functions taking into consideration the epidemiological revolution already underway, scientific progress and its practical applications in medicine together with the expectation for a better quality of life: we should be aiming at giving more life to years rather than more years to life. So it is in this context that hospitals can no longer be seen as the only, prevalent answer to health and care needs. An entirely innovative network of prevention services, diagnosis, cure and rehabilitation for the non self-sufficient must be provided, innovative also in the way it is supplied and managed, able to respond to the requirements of the patient in an appropriate and punctual fashion. Hospitalization of a patient suffering from dementia is always to be considered a critical event because of the protracted length of the stay, because of the increasing loss of vital functions in the patient together with the higher risk of complications including infection, falls and iatrogenic damage as well as an increased rate of mortality. The network of services provided is, therefore, fundamental. Centres specialised in the diagnosis of dementia (Memory clinics, outpatient dementia clinics, Alzheimer assessment units) are able to enlarge upon traditional services by providing the following within the protocols of diagnostic assessment and specific care programs: ~ early identification of the illness ~ more accurate identification of illnesses responsible for dementia ~ improvement of quality of life of both patient and caregiver ~ more accurate treatment Admission of dementia sufferers to special units has been shown to ~ reduce the frequency and intensity of behavioural disorders without, or with limited use of psychotropic drugs and physical constriction ~ delay functional loss ~ prevent complications including falls, under nourishment, immobility syndrome and delirium ~ improve the quality of the life of the patient, his family and caregivers Special dementia units base their specific mission on four central pillars ~ the staff ~ the environment ~ the programs and activities ~ the family

Due to the ageing of the population and the sharp increase in life expectancy, cancer diagnosed in the elderly population is rising and it has become an increasingly common problem in the Western world. According to the National Cancer Institute (NCI), for all cancers combined, the incidence rate in people 65 and over is 10 times higher than the rate for younger people, and the mortality is 16 times higher. The number of people in the United States who are 65 and older is expected to double by year 2030. A study published in 1999 in the New England Journal of Medicine found that while 63% of people in the general population age 65 or older had cancer, only 25% of patient in that age group were represented in clinical trials. The cut-off point at which an adult is considered ‘elderly’ has not been well defined. Usually, age 70 years is considered a reference point and is commonly used in clinical trials in oncology. One of the most intriguing aspects of ageing is how different the ageing process is from person to person; the basis for this variation is largely unknown. Clinical trials are generally used for the clinical development on new drugs/strategies. They are divided in three consecutive phases (I, II and III). The firsts two aim to characterize the action and to archive all knowledge necessary for better mange the experimental drugs. For all these reasons it is important to allow the inclusion of selected people without comorbidities that can interfere with the results. In contrast, phase III trials, considered the last step before registration, aim to demonstrate the efficacy/utility of the experimental drug/strategies on a number of patients representing the whole population of patients for a selected pathology. If it is tolerable to limited the participation of elderly patients, for their frailty, comorbidities or different metabolism, in the inclusion of phase I or dose finding and Phase II trials it is not acceptable to exclude them in the large phase III. Furthermore, it is now clear that Chronologic age cannot be used to predict the degree of comorbidity and of functional deterioration of the single individual up to age 85 at least; Performance status, which takes into account level of activity, ambulation, and ability to care for oneself-and the presence of comorbid conditions that may be exacerbated by treatment are more reliable criteria for clinical trial eligibility than age. In conclusion age alone should never make a patient ineligible for a trial.

The main neurological and neuropsychological disorders related to vascular attacks or strokes, chronic degenerative diseases such as Parkinson or Althaimer, incidental factors such as cranial cerebral trauma, cognitive decline of elderly, occupy a large part of the clinic of the speech therapist because of aspects of specific competence (communicative, cognitive-linguistic and oral functions). Deficits such as aphasia, dyspraxia, dementia, unravelling and other neuropsychological disorders are conditions that necessarily require a multidisciplinary intervention. Rehabilitation, with whole culture of disability related to it, is configured as the central aspect of doing rehabilitation that is related to the activity of the “person” in its entirety and complexity. The concept of “quality of care” presents the citizen as the center of interest for operators called to carry out an increasingly effective, timely, continuous, secure, appropriate practice( with respect to interpersonal communication centralizing the assisted ). The competence of the speech therapist consists of three components: the knowledge, the skills, the behavior; that “knowing how to act “on the whole that guarantees quality, that allows the identification of the best practice through the use of multidisciplinary and professional operational tools, that amends and aligns the behavior of professionals. The proper assessment with standardized instruments is also important, which bring forth the objectives in the short, medium and long term. In the field of neurological diseases in adults, unfortunately dramatic expectations are reported in the response to early therapy, particularly in our more disadvantaged region, with the phenomena of migration. In an alarming situation, there are, however, important experiences and centers of excellence also qualified by the virtuosity of many professionals who collaborate in quickly operational teams rehabilitation, also with the involvement of users’ associations. Aim of the seminary is to compare the forces still operative in the field spatial with other operations experienced in other regions, for the sharing of an uniform model of the “management of the elderly patient in our region” and consequent testing of validity. All repecting the Guidelines and recommendations resulting from the study of the scientific evidence that in the last two years involved authoritative representatives of the profession, scientific societies and organizations representing national.

Incorrect use of drugs may cause adverse events with serious consequences for patients. Adverse event (Adverse events) refers to an event related to the care process and which involves an injury to the patient, unintended and undesirable. Errors in drug therapy (also called error of therapy) are predictable events that may cause or lead to the inappropriate use of the drug or a threat to hospitalized patients, which may occur during the process of managing the drug for prevention and therefore should be considered by the whole management system of healthcare. These events, predictable and avoidable must be differentiated from the adverse drug reactions (ADR-Adverse Drug Reaction), linked to the drug itself and that are detected and evaluated by pharmacovigilance. The causes of errors in therapy are multifactorial and involve different health professionals. Basically 5 categories of error are recognized . • Error limitation • Clerical error / interpretation, • Error processing; • Error distribution • Administration error

When the possibility to submit the population to a screening program is considered, a series of parameters must be evaluated. Mainly four 1) prevalence of the pathology 2) the accuracy and the invasivity of the test used for the screening 3) the effectiveness of the necessary interventions to correct the pathology and the relative cost 4) the general economic commitment. Screening for abdominal aorta aneurysm corresponds to these criteria. The prevalence is elevated if the screening is turned to a selected population: for instance the incidence of the aneurysms of the abdominal aorta with greater dimensions than 3cm in males over sixty varies from 4% to 8%; if the subjects have vascular risk factors such as smoking and hypertension, the incidence increases (from two to five times); the prevalence in the women over 60 years old is just up to the 1,5%, but if considered in the subjects with familiar history or with multiple factors of vascular risk, it increases two or three times more. Ultrasounds are extremely accurate for the identification of the aneurysms of the abdominal aorta. The effectiveness of the treatment of the aneurysms of great dimensions is widely documented: the annual incidence of breakup and death because of aneurysms of the abdominal aorta larger than 5,5cm is equal to around16%, while the pre and post surgery mortality from aneurysms of these dimensions changes from 2% to 6%. Subjects with great aneurysms receive a definite benefit from the surgery. The identification of the "small aneurysms" moreover enables a suitable program of overseeing to continue until the possible correction. The general economic commitment is sustainable just because it is carried out with non-invasive investigations, without correlated consequences, repeatable and low cost, in contrast to the huge managerial costs of the treatment of the morbidity and the mortality correlated to the pathology. In conclusion enough data exist to hold effective screening for the search of aneurysms of the abdominal aorta in selected populations of subjects between the ages of 60 and 75 years old and that the cost for the society can be considered sustainable.

Background The median age of people living with HIV/AIDS is increasing: while efficacy of HAART has reduced mortality and prolonged the life of patients, low perception of risk and new late diagnosis are related to infection in an elderly age. Most new late diagnosis are performed in older hardly immuno-compromised subjects; the development of opportunistic diseases complicates management of therapeutic strategies (drugs interactions, cumulative toxicities, hypersensitivity reaction), mainly in presence of comorbities. Most randomized clinical trials with new drugs tend to avoid the enrolment of >50 years patients with proper comorbities of old age. So the few data we can analyze come from observational cohorts. While it is well defined that older people are more adherent to treatment and achieved an adequate control of HIV-RNA viremia more frequently than younger although an incomplete immuno-reconstitution, the development of drug toxicity with higher frequency is controversial. Elderly people are more frequent to show reduced hepatic and kidney function, frequently have comorbities requesting new therapies with a higher pill burden and higher risk of drug interactions. Most antiretroviral drugs are metabolized by cytochrome system. Treatment may accelerate mitochondrial dysfunction, nucleoside reverse transcriptase inhibitors may contribute to muscle wasting. Some typical side effects of HAART such as body change and facial lypoatrophy could be more evident when associated with natural facial changes occurring with aging and modify negatively self perceived body image and relationship capacity. Age was also shown to increase the risk of renal tubular disease in association with tenofovir. Age is one of the major cardio-vascular disease (CVD) risk factors and the inflammatory component may contribute to this increased risk despite HAART efficacy. The risk of myocardial infarctions is increased in older treated HIV+ patients: metabolic abnormalities are more frequent at baseline and the treatment could increase the cumulative risk of CVD.

Many literature studies show that almost one-third of elderly people who never have fallen before, have a fear of falling anyway. This rate increases in people that have fallen before. Fear plays a very important role in influencing the elderly person, causing a loss of independence in ADL. It’s important to evaluate the intensity of this loss to make educative interventions to support elderly people. A correct clinical evaluation is essential in taking care of elderly people. Pathologies and conditions that could facilitate falls have to be underlined during the clinical evaluation. Moreover, a multi-factorial evaluation of risks is very important in elderly people that have fallen before. To reduce the incidence of falls in the global population we must improve intervention that includes individual physical exercise programs that aim to increase strength, walking, balance, moves and climbing stairs. In high risk population it is necessary to add intervention in the house, infrastructure and furniture facilities. According to some scientific work, an unsafe house increases the risk of fall in 50%. of houses .

Contemporary research efforts have provided new insight into the processes underlying common hearing impairments include aging, genetic defects, and environmental stress such as excessive noise (NIHL) or chemotherapeutic drugs. The primary cause of hearing impairment involves the death of sensory cells (hair cells) and neurons in the cochlea which are irreplaceable in mammmals. Rehabilitative measures have traditionally been based on technical solutions: hearing aids and cochlear implants that can minimize the social impact of impairment, however the challenge for a successful hearing rehabilitation is definitely the prevention of cell death or regeneration of the sensorineural cells and neurons after cochlear damage. Addressing experimentally the first issue, we have shown that noise exposure and ototoxic drugs lead to apoptosis and by using neurotrophic factors and anti-apoptotic drugs as well as antioxidants, we have developed several therapeutic approaches of protection thus limiting the damage to the inner ear [1-3]. One of the most exciting areas involving hair-cell regeneration concerns regeneration of hair cells with the use of stem cells. The discovery of both adult and embryonic stem cells in the cochlea and evidence that they can be converted into hair cells raise hope for the development of stem-cell based treatment regimes to regenerate damaged cochlear components. Potential cell sources for inner ear transplantation include fetal dorsal root ganglion cells, neural progenitor cells, stem or progenitor cells isolated from inner ear, immortalized auditory neuroblast cells, embryonic stem cells (ESCs) and their derived neuronal cells, and marrow stromal cells and finally stem cells arising from amniotic fluid and placenta and olfactory progenitors [3,4]. Principal obstacles for the clinical application remain the effects of immunosuppression, induced-carcinogenesis and teratoma formation observed in the experimental models [3,4].

IntroductionPrescribing medications is often challenging in olderpeople with multiple chronic diseases. Non-adherenceto prescribed medications is among the most importantproblems, so that many people are not benefiting prop-erly from the drugs they are prescribed. Additionally,this also represents a costly waste for health services [1].Comorbidity and medication adherencePoor adherence to the prescribed therapy is highly pre-valent in older patients. Prevailing evidence suggeststhat age per se should not be considered as a risk factorfor poor adherence [2]. Lower educational level, lessaffluent economic status, cognitive/physical impairment,being assisted by foreign caregivers, and some chronicdiseases, such as chronic renal failure, are commonlyreported poorly modifiable correlates of non-adherence.Regimen complexity represents a to some extent modifi-able correlate [2]. Vision impairment, reduced manualdexterity, and patients’ perception of medication impor-tance may contribute to determine the relationshipbetween regimen complexity and non-adherence. Thereis minimal evidence to support interventions thatimprove medication adherence, and the effects of psy-chosocial interventions are largely unknown [1].Therapeutic goals in patients with multiplechronic conditionsPatients with multiple chronic conditions may vary inregard to their opinion about health outcomes such aslonger survival, prevention of disease-specific events,physical and cognitive function, and tolerable risk ofadverse drug reactions. A paradigmatic example of howchallenging this issue can be in clinical practice is thedifficulty in rational prescribing for older patients withmultiple chronic conditions and reduced life expectancy,when the likelihood of benefit and goals of care mustalso be considered in addition to satisfying the basicprinciples of optimal medication use in the elderly [3].

In considering the future role of the volatile anesthetic desflurane (Suprane®) in modern anesthesia practice, it is important to first consider the factors that contribute to a successful anesthetic in this increasingly day surgery orientation. An optimal anesthetic for the new millennium would include the following characteristics. 1.It should have a rapid and smooth onset of action. Efficiency is a particularly important factor in the ambulatory setting [1] 2.There should be a rapid recovery of cognitive functioning without clinically-significant discomfort. Postoperative pain is a major limiting factor in determining when a patient can be discharged home after surgery. 3.The absence of adverse effects such as nausea and vomiting is extremely important as emetic symptoms affect not only recovery times but also the incidence of unanticipated hospital admissions after day-case surgery, and patient satisfaction with their overall anesthetic experience. The low solubility of desflurane contributes to rapid emergence after anesthesia. Compared with sevoflurane, recovery times to eye opening, response to verbal commands, and orientation to person, place, and time have been found to be significantly shorter with desflurane [2-4]. When compared with the rapid, short-acting intravenous anesthetic propofol for induction and maintenance of anesthesia, desflurane displayed more favorable early recovery characteristics and facilitated the fast-tracking process [4,5]. Although patients receiving desflurane are less sedated in the early postoperative period than those receiving propofol, times to ambulation and discharge were similar. Given its favorable early recovery profile, desflurane would appear to be a useful alternative to isoflurane, sevoflurane and propofol for maintenance of anesthesia in a busy practice environment where early discharge is one of the major goals (e.g., cardiovascular, neurosurgery, day-case surgery). In conclusion, desflurane is the maintenance anesthetic agent of choice in a fast-tracking practice environment .

Femoral fractures have to be considered frequent causes of hospital admission for the elderly. These fractures are characterized by an uncertain prognosis in terms of both morbidity and mortality. The management of proximal femoral fractures in the elderly should be multidisciplinary, indeed the fall causing the fracture might follow a neurological, metabolic or cardiovascular disorder. Thus the treatment of the hidden disease is mandatory prior to fixing the fracture. Furthermore any unknown disease could significantly affect the health status of the patient increasing the operative risk. At admission to hospital accurate case history and physical examination have to be recorded. These fractures have to be surgically treated whenever it is possible. Indeed, the conservative treatment is a burden with a lot of complications. The early surgical treatment (within 24 hours from the injury) is effective to reduce complications and mortality. When choosing the appropriate surgical treatment, a preliminary distinction among the medial fractures (subcapital and transcervical) and the lateral ones (basal, intertrochanteric, subtrochanteric) is done. Medial fractures usually cause the interruption of the lateral epihisary blood vessels with consequent avascular necrosis of the femoral head thus requiring a hemi- or total arthroplasty. Lateral injuries should be treated preferably by reduction and internal fixation. The latter fractures may in turn be divided into two subgroups on the basis of integrity of the medial femoral region near the lesser trochanter (stable or unstable fractures). Stable fractures can be fixed with a dynamic hip screw, whereas unstable fractures are usually treated by means of a proximal femoral nail. Early surgical treatment allows resumption of walking, and a sudden recovery of the patient’s autonomy. However the mortality rate of patients with proximal femoral fracture is higher than the mortality rate of an age and sex-matched population.

Surgical resection of a malignant tumor in the head and neck regions quite often alters the complex act of swallowing. The goal of dysphagia rehabilitation is to return the patient’s swallowing to as near normal as possible. Although surgical procedures in the oral cavity, tongue, pharynx can affect preparation, control and transport of the bolus, functional disorders following laryngeal surgery are most common and severe. The larynx serves two critical functions during swallowing. First, it elevates anteriorly moving itself from the path of the bolus and contributing to the opening of the cricopharyngeal sphincter. Second, it protects the airways from inhaling acting as a three level valve -- the epiglottis, false vocal folds, and true vocal folds. Any surgical intervention affecting this closure will likely results in aspiration during the swallow. Patients undergoing total laryngectomy have few swallowing problems due to the separation of the airways from the esophagus. On the contrary both partial and reconstructive procedures can significantly affect both the respiratory and the alimentary function. Supraglottic laryngectomy can interfere with laryngeal elevation and sometimes with vocal fold adduction especially when the surgical procedure includes portions of the hyoid bone, base of tongue, aryepiglottic folds, or false vocal folds. Vertical laryngectomies reduce laryngeal closure. When the intervention is limited to one focal fold (cordectomy), the swallowing recovery is usually easily achieved. In frontolateral laringectomy the removal of tissues extends to the opposite vocal fold: in such cases swallowing recovery is still possible with a combination of increased effort and compensatory head posturing. In sub-total/reconstructive laryngectomies (Crico-Hyoido-Epiglotto-Pexy, CHEP and Crico-HyoidoPexy, CHP) the surgeon intends to create a neoglottis able to contracts during swallowing. In such cases critical factors in recovery of swallowing are considered the airway closure at the laryngeal entrance, the propulsive movement of the tongue base and its contact with the posterior pharyngeal wall. Two pernicious, life-threatening complications can slowly compromise the patient’s life aspiration, pneumonia and malnutrition. Diagnostic procedures of swallowing disorders are mainly represented by radiographic contrast examination and by fiberoptics endoscopic evaluation. Compensatory treatment strategies include postural changes, increasing sensory input, modifying volume, consistency and viscosity of foods. Rehabilitative procedures are designed to improve a range of movements of oral or pharyngo-laryngeal structures and increase their sensory-motor integration and timing. Exercises and manoeuvres designed to achieve such goals are considered in detail.