Cosmological lepton asymmetry with a nonzero mixing angle \theta_{13}

While the baryon asymmetry of the Universe is nowadays well measured by cosmological observations, the bounds on the lepton asymmetry in the form of neutrinos are still significantly weaker. We place limits on the relic neutrino asymmetries using some of the latest cosmological data, taking into account the effect of flavor oscillations. We present our results for two different values of the neutrino mixing angle \theta_{13}, and show that for large \theta_{13} the limits on the total neutrino asymmetry become more stringent, diluting even large initial flavor asymmetries. In particular, we find that the present bounds are still dominated by the limits coming from Big Bang Nucleosynthesis, while the limits on the total neutrino mass from cosmological data are essentially independent of \theta_{13}. Finally, we perform a forecast for COrE, taken as an example of a future CMB experiment, and find that it could improve the limits on the total lepton asymmetry approximately by up to a factor 6.6.Comment: 11 pages, 7 figures, 5 tables. v2: updated COrE specifications. v3: matches Phys. Rev. D accepted versio

PACAP and VIP increase the expression of myelin- markers in rat schwannoma cells

Defining the mechanisms regulating peripheral myelinogenesis in vitro remains a daunting task. One of the major complications is related to the inability of Schwann cells to produce myelin in vitro. However, a limited number of schwannoma cell lines are now being accepted as adequate model systems to study myelin expression since they present most of the features that characterize normal myelinating Schwann cells. PACAP and VIP peptides have been shown to play part in myelin maturation and synthesis in the central nervous system, but no data regarding their potential actions have been reported in the peripheral nervous system, particularly in vitro. In this study, we investigated the effects of both peptides on the expression of myelin-specific markers using the well-established rat RT4 schwannoma cell line. In addition, we endeavored to partly unravel the underlying molecular mechanisms involved. Expression of myelin-specific markers (MAG, MBP and MPZ, respectively) was assessed in cells grown either in the presence of normal serum (10% FBS) or serum starved and treated with or without 100nM PACAP or VIP. Effects of pharmacological blockade of PACAP and VIP receptors as well as of the main signaling pathways on the expression of myelin-markers were also determined. Our data shows that serum starvation (24h) was sufficient to induce a significant increase in the expression of myelin markers. This result was paralleled by a concurrent increase in the endogenous expression of both peptides, as well as of the non-specific PACAP/VIP receptor 2 (VPAC2), but not of VPAC1 or PAC1. Exogenous PACAP or VIP treatment further exacerbated starvation-driven expression of myelin markers, suggesting an active role of these peptides in myelin generation. Interestingly, stimulation with either peptides increased phosphorylation levels of Akt at Ser473 residue whereas they did not affect Erk1/2 activation. Treatment with the PAC1 receptor antagonist (PACAP6-38) had no effects on myelin markers expression while a non-specific VPAC receptor antagonist completely abrogated both starvation and/or starvation + peptide-induced expression of myelin markers. Similar effects were obtained in cell pretreated with the PKA inhibitor (H-89, 10μM), suggesting that peptide-driven induction of myelin markers occurs preferentially via the canonical PKA/Akt signaling pathway. In conclusion, the results here presented provide the basis for future studies on the role of these peptides in physiological myelination and in demyelinating pathologies such as Charcot-Mary-Tooth disease

Effectiveness of a home-based telerehabilitation system in patients after total hip arthroplasty: study protocol of a randomized controlled trial

Background: The demand for total hip arthroplasty (THA) is quickly rising given the escalating global incidence of hip osteoarthritis, and it is widely accepted that the post-surgery rehabilitation is key to optimize outcomes. The overall objective of this study is to evaluate the effectiveness of a new telerehabilitation solution, ReHub, for the physical function and clinical outcome improvement following THA. The specific aims of this manuscript are to describe the study design, protocol, content of interventions, and primary and secondary outcomes and to discuss the clinical rehabilitation impact of the expected experimental results. Methods/design: This prospective, randomized, controlled, parallel-group trial will include 56 patients who had undergone primary THA. Patients are randomized to a control group (standard rehabilitation during the 2-week stay in the rehabilitation clinic followed by 3 weeks of unsupervised home-based rehabilitation) or an experimental group (standard rehabilitation during the 2-week stay in the rehabilitation clinic followed by 3 weeks of home-based ReHub-assisted telerehabilitation). The primary outcome is physical performance assessed through the Timed Up-and-Go (TUG) test. Secondary outcomes include independence level, pain intensity, hip disability, hip range of motion, muscle strength, and patient's perception of clinical improvement. Discussion: Proving the clinical and cost-effectiveness of a home-based telerehabilitation program for physical and muscle function following THA could support its systematic incorporation in post-surgical rehabilitation protocols, which should be tailored to the individual and collective needs

Beneficial effects of PACAP in osteoarthritis cartilage. An “in vivo” and an “in vitro” morphological and biochemical study

Osteoarthritis (OA); the most common form of degenerative joint disease; is associated with variations in pro-inflammatory growth factor levels; inflammation and hypocellularity resulting from chondrocyte apoptosis (1). Pituitary adenylate cyclaseactivating polypeptide (PACAP) is a neuropeptide endowed with a range of trophic effects in several cell types; including chondrocytes (2). However; its role in OA has not been studied. To address this issue; we investigated whether PACAP expression is affected in OA cartilage obtained from experimentally-induced OA rat models; and then studied the effects of PACAP in isolated chondrocytes exposed to IL-1β in vitro to mimic the inflammatory milieu of OA cartilage. OA induction was established by histomorphometric and histochemical analyses. Changes in PACAP distribution in cartilage or its concentration in synovial fluid (SF) were assessed by immunohistochemistry and ELISA. Results showed that PACAP abundance in cartilage tissue and SF was high in healthy controls. OA induction decreased PACAP levels both in affected cartilage and SF. In vitro; PACAP prevented IL-1β-induced chondrocyte apoptosis; as determined by MTT assay; Hoechst staining and western blots of apoptotic-related proteins. These changes were also accompanied by decreased i-NOS and COX-2 levels; suggesting an anti-inflammatory effect. Altogether, these findings support a potential role for PACAP as a chondroprotective agent for the treatment of OA

Apoptosis activation in human carious dentin. An immunohistochemical study

he exact mechanisms and enzymes involved in caries progression are largely unclear (1). Apoptosis plays a key role in dentin remodelling related to damage repair; however, it is unclear whether apoptosis in decayed teeth is activated through the extrinsic or the intrinsic pathway (2-5). This in vivo immunohistochemical study explored the localization of TRAIL, DR5, Bcl-2 and Bax, the main proteins involved in apoptosis, in teeth with advanced caries. To evaluate TRAIL, DR5, Bcl-2 and Bax immunoexpressions twelve permanent carious premolars were included in paraffin and processed for immunohistochemistry. The results showed that TRAIL and DR5 were overexpressed in dentin and in pulp vessels and mononuclear cells; strong Bax immunostaining was detected in dilated dentinal tubules close to the lesion, and Bcl-2 staining was weak in some dentin areas under the cavity or altogether absent. These findings suggest that both apoptosis pathways are activated in dental caries. Further studies are required to gain insights into its biomolecular mechanisms and provide a contribution to therapeutic strategies

Enhanced expression of PACAP and of its high affinity receptor (PAC1) in the hippocampus and cerebral cortex of dopamine D3 knockout mice

Dopamine (DA) D3 receptor (D3R) is a pre-synaptic autoreceptor whose main role is to modulate DA release through a negative feedback regulatory loop. Immunolocalization studies from our research group have previously described that these receptors, despite being relatively low expressed in the central nervous system (CNS), may still be detected in less discreet brain regions normally associated to memory function, including the hippocampus and the cerebral cortex (1). Consistent with these findings, genetic deletion of the D3R in mice (D3R-/-) has been shown to have profound repercussions on the formation of new memories, especially fear associative memories (2). While it is now well-accepted that such pro-cognitive effects in these mice are presumably due to the increased DA bioavailability caused by the lack of autoreceptor function, it still remains to be established if there are molecular determinants directly or indirectly involved in ameliorating this specific type of associative learning. Pituitary adenylate cyclase-activating polypeptide (PACAP) is an endogenous peptide which is gaining scientific relevance because of its prominent “cognitive enhancer” function and the recent developments suggesting its active participation in the acquisition and consolidation of fear memories (3, 4). Based on these evidences, we thought it could have been of scientific relevance to assess whether PACAP expression, as well as that of its binding receptors, are affected in knockout mice showing this peculiar behavioural phenotype. We found that PACAP immunoreactivity (IR) was present at low levels in CA1 hippocampal subfield while moderate staining was observed in CA2-CA3 fields and in the dentate gyrus (DG) of wild-type (WT) mice. In sharp contrast, PACAP-IR was remarkably increased in all CA subfields, and particularly in CA1, CA3 and the DG regions of D3R-/- mice. Regarding the cerebral cortex (CX), PACAP expression was restricted to the V cortical layer in WT mice, whereas in D3R-/- mice, stained neurons were apparent both in the IV, V and VI cortical layers, with an overall increased staining score. In line with these findings, the expression of the PACAP-preferring PAC1 receptor, which was detectable only at moderate levels in the CA2 subfield of WTs, was enhanced in both CA2 and CA3 of D3R-/- mice. Interestingly, PAC1-IR was already present at moderate levels in the II-III cortical layers of WT mice, but genetic deletion of the D3R caused a remarkable spread of PAC1-stained neurons throughout all cortical layers, with the exception of layer I. We conclude that the absence or blockade of functional D3Rs from the brain enhances both PACAP and PAC1 receptor expression levels in the hippocampus and cerebral cortex. Considering the ameliorative role mediated by PACAP-PAC1 signalling in cognition, we infer that enhanced PACAP peptide and receptor expression may relate to the specific behavioural phenotype of these mice

Induction of tissue plasminogen activator (tPA) by pituitary adenylate cyclase-activating polypeptide (PACAP) in Schwann cell-like cultures

Peripheral nerve regeneration is dependent on the ability of regenerating neurites to migrate through cellular debris and altered extracellular matrix at the injury site, grow along the residual distal nerve sheath conduit, and reinnervate synaptic targets. In cell culture, growth cones of regenerating axons secrete PACAP, a peptide known to induce the expression of the protease tPA1. Here we tested the hypothesis that PACAP might also stimulate peripheral glial cells to release tPA to participate in nerve regeneration. More specifically, we addressed whether PACAP promoted the release and expression of tPA in the Schwann cell-like culture RT4-D6P2T, which shares biochemical and physical properties with Schwann cells2. We found that PACAP dose- and time-dependently stimulated tPA expression. Maxadilan, a potent PACAP receptor agonist, mimicked the effect of PACAP, whereas VIP, a PACAP-related peptide, produced only a moderate response. PACAP ability to stimulate tPA expression seemed to be dependent on the Akt/CREB signaling cascade, as inhibition using the PI3K/Akt blocker wortmannin or the TrkA/B inhibitor K252a both significantly dampened PACAP-evoked tPA expression. A similar effect was obtained in cells treated with the PACAP/VIP receptor antagonist PACAP6-38. We conclude that PACAP through the Akt/CREB intracellular pathway, acts as a potent inducer of tPA expression and release in Schwann-cell like cultures

A Role for Nuclear Phospholipase Cβ1 in Cell Cycle Control

Phosphoinositide signaling resides in the nucleus, and among the enzymes of the cycle, phospholipase C (PLC) appears as the key element both in Saccharomyces cerevisiae and in mammalian cells. The yeast PLC pathway produces multiple inositol polyphosphates that modulate distinct nuclear processes. The mammalian PLCbeta(1), which localizes in the nucleus, is activated in insulin-like growth factor 1-mediated mitogenesis and undergoes down-regulation during murine erythroleukemia differentiation. PLCbeta(1) exists as two polypeptides of 150 and 140 kDa generated from a single gene by alternative RNA splicing, both of them containing in the COOH-terminal tail a cluster of lysine residues responsible for nuclear localization. These clues prompted us to try to establish the critical nuclear target(s) of PLCbeta(1) subtypes in the control of cell cycle progression. The results reveal that the two subtypes of PLCbeta(1) that localize in the nucleus induce cell cycle progression in Friend erythroleukemia cells. In fact when they are overexpressed in the nucleus, cyclin D3, along with its kinase (cdk4) but not cyclin E is overexpressed even though cells are serum-starved. As a consequence of this enforced expression, retinoblastoma protein is phosphorylated and E2F-1 transcription factor is activated as well. On the whole the results reveal a direct effect of nuclear PLCbeta(1) signaling in G(1) progression by means of a specific target, i.e. cyclin D3/cdk4

Triple negative breast cancer: Shedding light onto the role of pi3k/akt/mtor pathway

Breast cancer is one of the most widespread carcinoma and one of the main causes of cancer-related death worldwide, especially in women aged between 35 and 75 years. Among the different subtypes, triple negative breast cancer (TNBC) is characterized by the total absence of the estrogen-receptor (ER) and progesteron-receptor (PR) expression as well as the lack of human epidermal growth factor receptor 2 (HER2) overexpression or gene amplification. These biological characteristics confer to TNBC a higher aggressiveness and relapse risk along with poorer prognosis compared to other subtypes. Indeed, 5-years survival rate is still low and almost all patients die, despite any adjuvant treatment which at moment represents the heading pharmacological approach. To date, several clinical trials have been designed to investigate the potential role of some molecular markers, such as VEGF, EGFR, Src and mTOR, for targeted treatments in TNBC. In fact, many inhibitors of the PI3K/AKT/mTOR pathway, frequently de-regulated in TNBC, are acquiring a growing interest and several inhibitors are in preclinical development or already in early phase clinical trials. In this Review, we investigated the role of the PI3K/AKT/mTOR pathway in TNBC patients, by summarizing the molecular features that led to the distinction of different histotypes of TNBC. Furthermore, we provided an overview of the inhibition mechanisms of the mTOR and PI3K/AKT signaling pathways, highlighting the importance of integrating biological and clinical data for the development of mTOR inhibitors in order to implement targeted therapies for TNBC patients