Characterization of Neuropeptide S (NPS) in view of its potential as a novel anxiolytic therapy for anxiety disorders

Anxiety disorders, such as posttraumatic stress disorder (PTSD), are characterized by a high prevalence and debilitating symptoms. However, the current first-line treatment for these conditions, which consists of selective serotonin reuptake inhibitors (SSRIs) and cognitive behavioral therapy, alongside symptomatic treatment with benzodiazepines, does not represent by far a functional solution for all affected patients. Therefore, identifying and characterizing novel candidates for alternative anxiolytic therapies are a crucial focus of psychiatric and neurobiological research. This study focuses on Neuropeptide S (NPS), a newly identified endogenous neuropeptide that has been shown to exert strong anxiolytic effects upon intracerebral injection in rodents. In an approach that combines basic research with incipient clinically relevant application, novel mechanisms and brain targets of NPS-mediated anxiolytic effects were identified, and a noninvasive application procedure also applicable in patients, namely the intranasal administration, was established for the first time for NPS in mouse models. In a first step, the feasibility of intranasal NPS delivery was established in mice using fluorophore-coupled NPS to allow intracerebral tracking. This method permitted for the first time tracking of intranasally applied substances within the brain at a single-cell resolution. These results not only proved the applicability of intranasal NPS administration in the mouse, but also allowed identification and characterization of hitherto undescribed cerebral NPS target cells, which were shown to be most likely exclusively neurons. Moreover, specific uptake of fluorescently labeled NPS in the hippocampus provided the first direct evidence linking this brain region, a well-known major player in the regulation of fear expression, to the NPS circuitry. Further investigation into the functional role of the hippocampus in NPS-elicited anxiolytic effects revealed that local microinjections of NPS into the ventral CA1 (vCA1) region are sufficient to elicit anxiolysis in C57BL6/N mice on the elevated plus maze (EPM). In a second step, behavioral and molecular effects of intranasal NPS treatment were characterized in C57BL/6N mice. Intranasal application of NPS was shown here to produce anxiolytic effects similar to those described by others after intracerebral injection. This finding represents the basis for the implementation of a future NPS-based therapy via nasal sprays in patients suffering from anxiety disorders. Furthermore, the molecular effects of NPS treatment on cerebral protein expression were examined here for the first time. This research led to identification of novel downstream targets of NPS-mediated regulation in the hippocampus and the prefrontal cortex. These new targets include proteins involved in the glutamatergic system and in synaptic plasticity, both of which are known to be dysregulated in anxiety disorders. Finally, the effects of intranasal NPS treatment, hitherto described only in non-pathological animal models, were examined for the first time in mouse models of anxiety disorders, namely the high anxiety behavior (HAB) mice and a mouse model of PTSD. In HAB mice, NPS treatment elicited anxiolytic effects similar to those observed in C57BL/6N mice. In the mouse model of PTSD, NPS counteracted disease-related changes in expression levels of hippocampal synaptic proteins. To sum up, this work expands the current state-of-knowledge concerning the molecular and mechanistic background of NPS-mediated anxiolysis by characterizing the role of the hippocampus in the NPS circuitry and by identifying novel downstream targets of NPS. The data on anxiolytic effects of intranasal NPS treatment especially in mouse models of anxiety disorders furthermore establishes the therapeutic potential of NPS as a novel anxiolytic treatment

Characterization of Neuropeptide S (NPS) in view of its potential as a novel anxiolytic therapy for anxiety disorders

Anxiety disorders, such as posttraumatic stress disorder (PTSD), are characterized by a high prevalence and debilitating symptoms. However, the current first-line treatment for these conditions, which consists of selective serotonin reuptake inhibitors (SSRIs) and cognitive behavioral therapy, alongside symptomatic treatment with benzodiazepines, does not represent by far a functional solution for all affected patients. Therefore, identifying and characterizing novel candidates for alternative anxiolytic therapies are a crucial focus of psychiatric and neurobiological research. This study focuses on Neuropeptide S (NPS), a newly identified endogenous neuropeptide that has been shown to exert strong anxiolytic effects upon intracerebral injection in rodents. In an approach that combines basic research with incipient clinically relevant application, novel mechanisms and brain targets of NPS-mediated anxiolytic effects were identified, and a noninvasive application procedure also applicable in patients, namely the intranasal administration, was established for the first time for NPS in mouse models. In a first step, the feasibility of intranasal NPS delivery was established in mice using fluorophore-coupled NPS to allow intracerebral tracking. This method permitted for the first time tracking of intranasally applied substances within the brain at a single-cell resolution. These results not only proved the applicability of intranasal NPS administration in the mouse, but also allowed identification and characterization of hitherto undescribed cerebral NPS target cells, which were shown to be most likely exclusively neurons. Moreover, specific uptake of fluorescently labeled NPS in the hippocampus provided the first direct evidence linking this brain region, a well-known major player in the regulation of fear expression, to the NPS circuitry. Further investigation into the functional role of the hippocampus in NPS-elicited anxiolytic effects revealed that local microinjections of NPS into the ventral CA1 (vCA1) region are sufficient to elicit anxiolysis in C57BL6/N mice on the elevated plus maze (EPM). In a second step, behavioral and molecular effects of intranasal NPS treatment were characterized in C57BL/6N mice. Intranasal application of NPS was shown here to produce anxiolytic effects similar to those described by others after intracerebral injection. This finding represents the basis for the implementation of a future NPS-based therapy via nasal sprays in patients suffering from anxiety disorders. Furthermore, the molecular effects of NPS treatment on cerebral protein expression were examined here for the first time. This research led to identification of novel downstream targets of NPS-mediated regulation in the hippocampus and the prefrontal cortex. These new targets include proteins involved in the glutamatergic system and in synaptic plasticity, both of which are known to be dysregulated in anxiety disorders. Finally, the effects of intranasal NPS treatment, hitherto described only in non-pathological animal models, were examined for the first time in mouse models of anxiety disorders, namely the high anxiety behavior (HAB) mice and a mouse model of PTSD. In HAB mice, NPS treatment elicited anxiolytic effects similar to those observed in C57BL/6N mice. In the mouse model of PTSD, NPS counteracted disease-related changes in expression levels of hippocampal synaptic proteins. To sum up, this work expands the current state-of-knowledge concerning the molecular and mechanistic background of NPS-mediated anxiolysis by characterizing the role of the hippocampus in the NPS circuitry and by identifying novel downstream targets of NPS. The data on anxiolytic effects of intranasal NPS treatment especially in mouse models of anxiety disorders furthermore establishes the therapeutic potential of NPS as a novel anxiolytic treatment

NON-COMPETITION CLAUSES IN COMMERCIAL CONTRACTS

We begin with an analysis of areas where rivalry between economic agents can not show (any act of competition committed in this area drawing the liability of the author), we will then analyze competition in relations between the trader and servant or other employees and continue with the analysis of the legal ban on competition in the limited liability companies and joint stock companies. So, the relevant provisions of Law 31/1990 are reviewed, views of legal doctrine and practice of judicial rulings on the nature and purpose of the relevant provisions referred to, their scope, applicability of statutory prohibition against competition in the profile activity of the company, the prohibition in the liquidation phase, procedural methods which can cover damage caused to the creditor’s violated rights, as well as statute of limitations for the right to action and prescription

L’opposition parlementaire en Roumanie postcommuniste: 1990-2004

To what extent the parliamentary activity of the Romanian political parties reflects the division line between "power" and "opposition", as it is defined by the alliances that shape the composition of the government and of its parliamentary support? According to data analysis, the most straightforward answer to this question should be: the parliamentary activity of the political parties reflects only moderately the division line between "power" and "opposition". There is a clear and frequent support given to the government and to its policies, by the parties that do not belong to the parliamentary majority. The division line between the two camps is crossed form both sides but, in most of the cases, the opposition parties are the ones who cross it. Nevertheless, the data gathered do not offer enough arguments neither to change completely the image of the "power" and "opposition" camps, as it was publicly perceived during the whole period, nor to nominate certain parties that have constantly transgressed the division line between the two camps

Romanian Aromatic and Medicinal Plants: From Tradition to Science

From ancient times, plants have been used by humans for food, fodder, fibre and medicinal purposes. Several plants were empirically considered as treatments for a large array of illness and medical conditions. Each community had specific natural remedies, based on the geographical area, environmental conditions and other factors. Thus, the use of plants can be considered as part of the intangible cultural heritage of each community. In the geographical area of today’s Romania, the ancient inhabitants, Dacians, had very good knowledge regarding the use of plants for medicinal purposes, as presented by several historical sources. The present work describes protocols for the extraction and purification of natural extracts, analytical characterisation, in vitro and in vivo evaluation of their potential applications as well as some practical examples of their application on selected Romanian native medicinal and aromatic plants. The presented results offer scientific support to their traditional use, suggesting in the same time some modern applications, for example in the nanotechnology field

Quantifying the economic survive across the EU using Markov probability chains

The multiple global crisis has made the economies of the world’s countries, including EU’s economy, vulnerable through the downgrading of the pandemic and the subsequent outbreak of geo-political conflict. These two events had the effect of decelerating the European economy and increasing the poverty level of the population, even that these developments are weaker than in rest of the world. The main objective of the present scientific approach is to identify a risk function based on Markov probability chains and to assess the possibilities of economic recovery through a package of policies structured over different time horizons. The used methods consist of meta-analysis, statistical analysis and geo-spatial and temporal modelling. The results of the study capture the integrated developments of risk-generating macroeconomic elements such as inflation, unemployment, public debt growth in a regionally segregated manner. These elements are useful for supranational decision-makers to increase the economic survival rate after multiple shocks through our proposed policy package. First published online 14 March 202