Der Einfluss von Clomethiazol, Clozapin und Valproinsaeure auf die Expression von NPY und seinem Rezeptorsystem.

Summary Background and aims Neuropeptide Y is a widely expressed peptide in the central and peripheral nervous system and has been found to be associated with many physiological functions. Acting via four different G-protein coupled receptors (Y1, Y2, Y4, Y5) it is involved in feeding behaviour and the pathology of obesity, anxiety, blood pressure, addiction and convulsion. Psychotropic drugs are designed to modulate those functions, even though little is known about the exact effects on cellular pathways. Gain of weight, epileptic or antiepileptic effects, sedation and anxiolysis are among the effects and side effects of valproic acid, clomethiazole and clozapine, suggesting that these drugs effect NPY and its receptor-system. The aim of the present study was to investigate the effects of valproic acid, clomethiazole and clozapine on gene expression of NPY and its receptors, which is a step towards understanding the drugs‘ mechanisms of action. Material and methods Human neuroblastoma cells, SH-SY5Y (SY5Y) and human embryonic kidney cells, HEK-293 (HEK) were stimulated with the above-named substances for 4, 24 and 48 hours. The employed drug-concentrations for incubation were determined using cytotoxicity-tests. The gene expression level of NPY and its receptors was measured by relative quantification using real-time dye-based PCR and the protein expression was analysed for selected genes using western blots. Results Stimulation with valproic acid showed a significant increase of gene expression of NPY, Y1, Y2 and Y5 in HEK-cells. Induction of Y1 was also seen in SY5Y-cells. Clomethiazole showed various effects on the NPY system, inhibiting the expression of Y2 and Y5 in HEK but increasing the expression of NPY and Y1 in SY5Y and also inhibiting Y2. Stimulation with clozapine showed only a decrease in expression for Y2 in HEK and no other significant results. Conclusions The results give evidence, that the effects and side effects of valproic acid (sedation, antiepileptic effects, anxiolysis, gain of weight) are correlated with modulation in gene-expression of NPY and its receptors. It could be shown that clozapine can only unfold its physiological effects (sedation, gain of weight, epileptic effects), if at all via affecting the NPY-system, through repression of Y2. Since the data of clomethiazole ist partly contrary to its physiological effects (sedation, anticonvulsion), the detectable and significant changes in gene-expression of the NPY-system are probably covered from other, physiologically stronger mechanisms. However, there are interesting ways of interpreting the repression of Y2 for the anxiolytic and also antiepileptic effects of clomethiazole and for the physiological effects of clozapine.Zusammenfassung Hintergrund und Ziele Neuropeptid Y (NPY) ist ein weit verbreitetes und viel erforschtes Neuropeptid im zentralen und peripheren Nervensystem. Über vier verschiedene, G-Protein-gekoppelte Rezeptoren (Y1, Y2, Y4, Y5) ist es als Transmitter an der Regulation vieler physiologischer Funktionen wie Essverhalten und Entstehung von Übergewicht, Angstverhalten, Krampfentwicklung, Blutdruck und Entstehung von Abhängigkeit beteiligt. Die Modulation dieser Funktionen ist Effekt vieler psychopharmakologischer Behandlungen, wobei der genaue Wirkmechanismus der Medikamente häufig noch nicht vollständig aufgeklärt ist. Gewichtszunahme, antiepileptische Wirkung oder Senkung der Krampfschwelle, sowie Sedation und Anxiolyse, zählen zu den erwünschten und unerwünschten Wirkungen von Valproinsäure, Clomethiazol und Clozapin, und die Vermutung liegt nahe, dass diese Medikamente Teile ihrer Wirkungen über Modulation von Neuropeptid Y in Verbindung mit seinen vier Rezeptoren entfalten. So ist es das Ziel dieser Arbeit, herauszufinden, ob sich die Genexpression von Neuropeptid Y und seinen Rezeptoren unter Stimulation mit Valproinsäure, Clomethiazol und Clozapin verändert und so zur Aufklärung der molekularen Wirkmechanismen dieser Medikamente beitragen. Material und Methoden Humane Neuroblastomzellen, SH-SY5Y (SY5Y) und humane, embryonale Nierenzellen, HEK-293 (HEK) wurden in Zellkulturexperimenten mit oben genannten Substanzen für je 4, 24 und 48 Stunden stimuliert. Die hierfür geeigneten Stimulationskonzentrationen wurden zuvor mittels eines Zytotoxizitätstests bestimmt. Die Ermittlung des Expressionsniveaus für NPY und seine Rezeptoren erfolgte relativ im Vergleich zu unbehandelten Kontrollen durch eine quantitative PCR auf Ebene der Gentranskription und teilweise folgte im Western-Blot die Analyse der nachfolgenden Translation. Ergebnisse Nach Stimulation mit Valproinsäure zeigten sich in der HEK-Zelllinie signifikante Anstiege der Expression von NPY, Y1, Y2 und Y5. Die Induktion von Y1 ließ sich auch in den SY5Y Zellen zeigen. Clomethiazol zeigte variable Beeinflussung des NPY-Systems; während in den HEK-Zellen eine Inhibition von Y2 und Y5 nachweisbar war, führte es bei den Zellen der Linie SY5Y zu einer Induktion von NPY selbst und Y1 und ebenfalls zu einer Inhibition von Y2. Nach Stimulation mit Clozapin konnte bei der Zelllinie HEK eine verminderte Expression von Y2 festgestellt werden, alle anderen Untersuchungen zeigten keine signifikanten Veränderungen. Schlussfolgerungen Die Ergebnisse unterstützen die Vermutung, dass erwünschte und unerwünschte Wirkungen von Valproinsäure (Sedation, Antikonvulsion, Anxiolyse, Gewichtszunahme) durch Veränderung der Expression von NPY und seinen Rezeptoren mitverursacht werden. Im Gegensatz dazu konnte gezeigt werden, dass Clozapin seine Wirkungen (Sedierung, Gewichtszunahme, Senkung der Krampfschwelle), wenn sie überhaupt durch Veränderungen im NPY-System hervorgerufen werden, nur über die Beeinflussung von Y2 entfalten kann. Die Ergebnisse für Clomethiazol legen nahe, dass nachweisbare Veränderungen der Expression von NPY und seinen Rezeptoren teilweise durch physiologisch stärkere, andere Wirkungen überdeckt werden, da die nachgewiesenen zellulären Veränderungen partiell im Gegensatz zur antikonvulsiven und sedierenden Wirkung stehen. Insgesamt ergibt die Repression von Y2 interessante Interpretationsmöglichkeiten bezüglich der anxiolytischen und auch antikonvulsiven Wirkung von Clomethiazol und der physiologischen Wirkungen von Clozapin

Research for transition: Europeans deserve a better agricultural and food policy

Research is a key element in the exploration of new pathways in farming systems. Organic farming relies on specific methods and a strict regulation. By design, organic farming harmonizes the environmental and productive dimensions of farming systems. The report covers four sections. 1. Funding of organic farming research is low both at EU and national levels, but at the EU and national levels, statistics on the financial support to the different models of agriculture are neither precise nor comprehensive. This lack of transparency impairs any comparative analysis. 2. Several countries have specific programmes for organic farming research The total amounts of money are limited but in most cases the programmes are multi-annual and help to build long-term expertise for the sector. Countries with long-term programmes include Denmark, France, Germany and Sweden 3. Organic farming provides better answer to sustainability challenges than conventional farming. Funding of organic farming research is important because organic farming represents an efficient pathway to sustainable agriculture. A comparison of organic and conventional farming for the different dimensions of sustainability has been compiled based on scientific publications. This assessment does not claim to be fully comprehensive in all areas but it may serve to illustrate he potential of organic farming. 4. Inspiring case studies. By design, organic farming is multi-functional and based on an ecosystem approach rather than the use of artificial inputs that boost production. This is also reflected in the organisation of knowledge exchanges. Most organic farmers are in favour of a participatory vision of research, with active exchange of experience between scientists and practitioners, a collective assessment of problems and a co-design of solutions. Programmes such as the European Innovation Partnership are in line with this research and innovation process. Experience in organic farming shows the potential of such an approach. Case studies at meso and micro levels illustrate new ways of producing knowledge in a participatory way

From RNA-seq to large-scale genotyping - genomics resources for rye (Secale cereale L.)

<p>Abstract</p> <p>Background</p> <p>The improvement of agricultural crops with regard to yield, resistance and environmental adaptation is a perpetual challenge for both breeding and research. Exploration of the genetic potential and implementation of genome-based breeding strategies for efficient rye (<it>Secale cereale </it>L.) cultivar improvement have been hampered by the lack of genome sequence information. To overcome this limitation we sequenced the transcriptomes of five winter rye inbred lines using Roche/454 GS FLX technology.</p> <p>Results</p> <p>More than 2.5 million reads were assembled into 115,400 contigs representing a comprehensive rye expressed sequence tag (EST) resource. From sequence comparisons 5,234 single nucleotide polymorphisms (SNPs) were identified to develop the Rye5K high-throughput SNP genotyping array. Performance of the Rye5K SNP array was investigated by genotyping 59 rye inbred lines including the five lines used for sequencing, and five barley, three wheat, and two triticale accessions. A balanced distribution of allele frequencies ranging from 0.1 to 0.9 was observed. Residual heterozygosity of the rye inbred lines varied from 4.0 to 20.4% with higher average heterozygosity in the pollen compared to the seed parent pool.</p> <p>Conclusions</p> <p>The established sequence and molecular marker resources will improve and promote genetic and genomic research as well as genome-based breeding in rye.</p

Effects of 3-D thermal radiation on the development of a shallow cumulus cloud field

We investigate the effects of thermal radiation on cloud development in large-eddy simulations (LESs) with the UCLA-LES model. We investigate single convective clouds (driven by a warm bubble) at 50m horizontal resolution and a large cumulus cloud field at 50 and 100m horizontal resolutions. We compare the newly developed 3-D Neighboring Column Approximation with the independent column approximation and a simulation without radiation and their respective impact on clouds. Thermal radiation causes strong local cooling at cloud tops accompanied by a modest warming at the cloud bottom and, in the case of the 3-D scheme, also cloud side cooling. 3-D thermal radiation causes systematically larger cooling when averaged over the model domain. In order to investigate the effects of local cooling on the clouds and to separate these local effects from a systematically larger cooling effect in the modeling domain, we apply the radiative transfer solutions in different ways. The direct effect of heating and cooling at the clouds is applied (local thermal radiation) in a first simulation. Furthermore, a horizontal average of the 1-D and 3-D radiation in each layer is used to study the effect of local cloud radiation as opposed to the domain-averaged effect. These averaged radiation simulations exhibit a cooling profile with stronger cooling in the cloudy layers. In a final setup, we replace the radiation simulation by a uniform cooling of 2.6 K day(-1). To focus on the radiation effects themselves and to avoid possible feedbacks, we fixed surface fluxes of latent and sensible heat and omitted the formation of rain in our simulations. Local thermal radiation changes cloud circulation in the single cloud simulations, as well as in the shallow cumulus cloud field, by causing stronger updrafts and stronger subsiding shells. In our cumulus cloud field simulation, we find that local radiation enhances the circulation compared to the averaged radiation applications. In addition, we find that thermal radiation triggers the organization of clouds in two different ways. First, local interactive radiation leads to the formation of cell structures;later on, larger clouds develop. Comparing the effects of 3-D and 1-D thermal radiation, we find that organization effects of 3-D local thermal radiation are usually stronger than the 1-D counterpart. Horizontally averaged radiation causes more clouds and deeper clouds than a no radiation simulation but, in general less-organized clouds than in the local radiation simulations. Applying a constant cooling to the simulations leads to a similar development of the cloud field as in the case of averaged radiation, but less water condenses overall in the simulation. Generally, clouds contain more liquid water if radiation is accounted for. Furthermore, thermal radiation enhances turbulence and mixing as well as the size and lifetime of clouds. Local thermal radiation produces larger clouds with longer lifetimes. The cloud fields in the 100 and 50 m resolution simulations develop similarly;however, 3-D local effects are stronger in the 100m simulations which might indicate a limit of our 3-D radiation parameterization

GeneTrail 3: advanced high-throughput enrichment analysis

We present GeneTrail 3, a major extension of our web service GeneTrail that offers rich functionality for the identification, analysis, and visualization of deregulated biological processes. Our web service provides a comprehensive collection of biological processes and signaling pathways for 12 model organisms that can be analyzed with a powerful framework for enrichment and network analysis of transcriptomic, miRNomic, proteomic, and genomic data sets. Moreover, GeneTrail offers novel workflows for the analysis of epigenetic marks, time series experiments, and single cell data. We demonstrate the capabilities of our web service in two case-studies, which highlight that GeneTrail is well equipped for uncovering complex molecular mechanisms. GeneTrail is freely accessible at: http://genetrail.bioinf.uni-sb.de

Genomic prediction within and across maize landrace derived populations using haplotypes

Genomic prediction (GP) using haplotypes is considered advantageous compared to GP solely reliant on single nucleotide polymorphisms (SNPs), owing to haplotypes’ enhanced ability to capture ancestral information and their higher linkage disequilibrium with quantitative trait loci (QTL). Many empirical studies supported the advantages of haplotype-based GP over SNP-based approaches. Nevertheless, the performance of haplotype-based GP can vary significantly depending on multiple factors, including the traits being studied, the genetic structure of the population under investigation, and the particular method employed for haplotype construction. In this study, we compared haplotype and SNP based prediction accuracies in four populations derived from European maize landraces. Populations comprised either doubled haploid lines (DH) derived directly from landraces, or gamete capture lines (GC) derived from crosses of the landraces with an inbred line. For two different landraces, both types of populations were generated, genotyped with 600k SNPs and phenotyped as lines per se for five traits. Our study explores three prediction scenarios: (i) within each of the four populations, (ii) across DH and GC populations from the same landrace, and (iii) across landraces using either DH or GC populations. Three haplotype construction methods were evaluated: 1. fixed-window blocks (FixedHB), 2. LD-based blocks (HaploView), and 3. IBD-based blocks (HaploBlocker). In within population predictions, FixedHB and HaploView methods performed as well as or slightly better than SNPs for all traits. HaploBlocker improved accuracy for certain traits but exhibited inferior performance for others. In prediction across populations, the parameter setting from HaploBlocker which controls the construction of shared haplotypes between populations played a crucial role for obtaining optimal results. When predicting across landraces, accuracies were low for both, SNP and haplotype approaches, but for specific traits substantial improvement was observed with HaploBlocker. This study provides recommendations for optimal haplotype construction and identifies relevant parameters for constructing haplotypes in the context of genomic prediction

Mobile Air Quality Studies (MAQS) - an international project

Due to an increasing awareness of the potential hazardousness of air pollutants, new laws, rules and guidelines have recently been implemented globally. In this respect, numerous studies have addressed traffic-related exposure to particulate matter using stationary technology so far. By contrast, only few studies used the advanced technology of mobile exposure analysis. The Mobile Air Quality Study (MAQS) addresses the issue of air pollutant exposure by combining advanced high-granularity spatial-temporal analysis with vehicle-mounted, person-mounted and roadside sensors. The MAQS-platform will be used by international collaborators in order 1) to assess air pollutant exposure in relation to road structure, 2) to assess air pollutant exposure in relation to traffic density, 3) to assess air pollutant exposure in relation to weather conditions, 4) to compare exposure within vehicles between front and back seat (children) positions, and 5) to evaluate "traffic zone"- exposure in relation to non-"traffic zone"-exposure. Primarily, the MAQS-platform will focus on particulate matter. With the establishment of advanced mobile analysis tools, it is planed to extend the analysis to other pollutants including including NO2, SO2, nanoparticles, and ozone

How managers can build trust in strategic alliances: a meta-analysis on the central trust-building mechanisms

Trust is an important driver of superior alliance performance. Alliance managers are influential in this regard because trust requires active involvement, commitment and the dedicated support of the key actors involved in the strategic alliance. Despite the importance of trust for explaining alliance performance, little effort has been made to systematically investigate the mechanisms that managers can use to purposefully create trust in strategic alliances. We use Parkhe’s (1998b) theoretical framework to derive nine hypotheses that distinguish between process-based, characteristic-based and institutional-based trust-building mechanisms. Our meta-analysis of 64 empirical studies shows that trust is strongly related to alliance performance. Process-based mechanisms are more important for building trust than characteristic- and institutional-based mechanisms. The effects of prior ties and asset specificity are not as strong as expected and the impact of safeguards on trust is not well understood. Overall, theoretical trust research has outpaced empirical research by far and promising opportunities for future empirical research exist

Evaluation of Concomitant Systemic Treatment in Older Adults With Head and Neck Squamous Cell Carcinoma Undergoing Definitive Radiotherapy

IMPORTANCE The number of older adults with head and neck squamous cell carcinoma (HNSCC) is increasing, and these patients are underrepresented in clinical trials. It is unclear whether the addition of chemotherapy or cetuximab to radiotherapy is associated with improved survival in older adults with HNSCC. OBJECTIVE To examine whether the addition of chemotherapy or cetuximab to definitive radiotherapy is associated with improved survival in patients with locoregionally advanced (LA) HNSCC. DESIGN, SETTING, AND PARTICIPANTS The Special Care Patterns for Elderly HNSCC Patients Undergoing Radiotherapy (SENIOR) study is an international, multicenter cohort study including older adults (≥65 years) with LA-HNSCCs of the oral cavity, oropharynx/hypopharynx, or larynx treated with definitive radiotherapy, either alone or with concomitant systemic treatment, between January 2005 and December 2019 at 12 academic centers in the US and Europe. Data analysis was conducted from June 4 to August 10, 2022. INTERVENTIONS All patients underwent definitive radiotherapy alone or with concomitant systemic treatment. MAIN OUTCOMES AND MEASURES The primary outcome was overall survival. Secondary outcomes included progression-free survival and locoregional failure rate. RESULTS Among the 1044 patients (734 men [70.3%]; median [IQR] age, 73 [69-78] years) included in this study, 234 patients (22.4%) were treated with radiotherapy alone and 810 patients (77.6%) received concomitant systemic treatment with chemotherapy (677 [64.8%]) or cetuximab (133 [12.7%]). Using inverse probability weighting to attribute for selection bias, chemoradiation was associated with longer overall survival than radiotherapy alone (hazard ratio [HR], 0.61; 95% CI, 0.48-0.77; P < .001), whereas cetuximab-based bioradiotherapy was not (HR, 0.94; 95% CI, 0.70-1.27; P = .70). Progression-free survival was also longer after the addition of chemotherapy (HR, 0.65; 95% CI, 0.52-0.81; P < .001), while the locoregional failure rate was not significantly different (subhazard ratio, 0.62; 95% CI, 0.30-1.26; P = .19). The survival benefit of the chemoradiation group was present in patients up to age 80 years (65-69 years: HR, 0.52; 95% CI, 0.33-0.82; 70-79 years: HR, 0.60; 95% CI, 0.43-0.85), but was absent in patients aged 80 years or older (HR, 0.89; 95% CI, 0.56-1.41). CONCLUSIONS AND RELEVANCE In this cohort study of older adults with LA- HNSCC, chemoradiation, but not cetuximab-based bioradiotherapy, was associated with longer survival compared with radiotherapy alone