Functional consequences of nociceptin receptor activation

Nociceptin orphanin FQ (N/OFQ) is the 17 amino acid endogenous ligand for the Gi-coupled N/OFQ-receptor (NOP). In vivo administration produces a wide range of physiological responses including; analgesia, hyperalgesia and anti-opioid actions.;In a series of in vitro assays including [leucyl -3H]N/OFQ binding, GTPgamma[35S] binding and inhibition of cAMP formation the following linked studies were performed; (1)N/OFQ structure-activity relationships (SAR) in cells (CHO) stably expressing human NOP (2)evaluation of receptor density on efficacy using the ecdysone inducible expression system and native tissues, (3)an investigation of NOP/G-protein coupling efficiency.;SAR studies can be summarized as combining arginine14, lysine15 repeat in N/OFQ (increase affinity/potency) with C-terminal [F/G]N/OFQ(1--13)NH2([F/G]) and [Nphe1]N/OFQ(1--13)NH 2([Nphe1]) modifications (reduce/eliminate efficacy). Arg14/Lys15 increased the affinity (pKi:10.31--11.16) and potency (pEC50:8.98--9.85) of N/OFQ. [F/G] and [Nphe 1] reduced the efficacy of N/OFQ from 1.0 to 0.44 and 0. Combination of Arg14/Lys15 and [Nphe1] in UFP-101 produced the highest affinity peptide antagonist available (pA 2:9.13, [Nphe1] alone:7.54). Using the ecdysone inducible expression system it was possible to vary the efficacy of [F/G], between antagonist (alpha:0 upstream/low density), partial agonist (alpha:0.3--0.75) and full agonist (alpha:1). In the mouse vas deferens and colon [F/G] displayed varying degrees of partial agonism possibly indicating differences in NOP receptor density. The binding of GDP and GTP in CHOhNOP was to high and low affinity sites. The fraction of GDP binding to the high affinity site was reduced by N/OFQ (77%, under basal to 32%). The reduction in high affinity binding of GDP appeared dependent on the efficacy of the ligand.;This thesis has identified several new peptides of varying efficacy/potency for use in defining the pathophysiological role(s) of N/OFQ-NOP. Efficacy is not just a property of the ligand but of the assay systems and its input receptor density. A cautious approach to new ligands characterized at a single endpoint is advocated

BSF 1102

Detailed alignment and annotation information on 1959 filtered chimeric transcripts from 55 samples analyzed in the study. Each chimeric transcript is represented by a unique ID in the first column. Structural and functional classification (as described in the text) information is presented in columns S, T and U. Cells in the gene name columns (‘geneName1’ and ‘geneName2’) with value “none” represent gene-desert regions. The first sheet in the excel file contains the data columns and a key describing the data is on the second excel sheet. (XLSX 684 kb

Comparison of long-term intents of education and development in regions of Czech republic

In this work there is accomplished a comparison of long-term intentions of education and development of educational system in four regions of Czech republic - Jihocesky, Karlovarsky, Plzensky, Stredocesky. It's with aim to find how much are these intentions identical or different in chosen educational areas. The work compares fields - parity of opportunities to education, to support of foreign languages and informational and comunication technologies, also area of pedagogical workers and strategy of development individual levels of education according to demographical progress. At the same time, there is made the comparison of these long-term intentions of regions with long-term intentions of education and development of Czech republic

Hot Topics in Opioid Pharmacology: Mixed and Biased Opioids.

Analgesic design and evaluation has been driven by the desire to create high affinity, high selectivity MOP (mu;µ) agonists. Such ligands are the mainstay of current clinical practice and include morphine and fentanyl. Advances in this sphere have come from designing pharmacokinetic advantage; rapid metabolism for remifentanil. These produce analgesia but also the adverse effect profile that currently defines this drug class; namely ventilatory depression, tolerance and abuse liability. The MOP receptor is part of a family and there are significant functional interactions between other members of the family (DOP;delta;δ, KOP;kappa;κ and NOP;nociceptin/orphanin FQ). Experimentally MOP agonism and DOP antagonism produced antinociception (animals) with no tolerance and low doses of MOP and NOP ligands synergize to antinociceptive advantage. In this latter context lack of effect of NOP agonists on ventilation is an additional advantage. Recent development has been to move towards low selectivity multifunctional ‘mixed ligands’ (e.g., Cebranopadol) or ligand mixtures (e.g., Targinact). Moreover, the observation that β-arrestin coupling underlies the side effect profile for MOP ligands (from knockout animal studies) led to the discovery of biased (to G-protein and away from β-arrestin) MOP ligands (e.g., oliceridine). There is sufficient excitement in the opioid field to suggest that opioid analgesics without significant side effects may be on the horizon and the ‘opioid holy grail’ might be in reach

Additional file 7: of Snail-induced epithelial-to-mesenchymal transition of MCF-7 breast cancer cells: systems analysis of molecular changes and their effect on radiation and drug sensitivity

List of differentially expressed genes between MCF-7-Snail and MCF-7-Control cells known to MetaCore knowledgebase to be directly transcriptionally regulated by c-Myc. (XLS 610 kb

Cannabinoid receptor expression in the bladder is altered in detrusor overactivity

Introduction: Immunohistochemical (IHC) evidence shows that cannabinoid receptors (CB) are expressed in human bladders and cannabinoid agonists are known to inhibit detrusor contractility. However, the mechanism for this inhibition remains unknown. In addition, the role of CB in detrusor overactivity (DO) is under-investigated. The aim of this study was to compare CB expression in normal and DO human bladders and to further characterise these receptors. Methods: Polymer chain reaction (PCR) was used to detect differences in CB transcripts in bladder samples. Differences in CB protein expression was assessed by IHC. Immunofluorescence (IF) was used to evaluate co-localisation of CB with nerve fibres. Receptor density and binding affinity were measured using the cannabinoid radioligand [3H]-CP-55,940. Results: There were higher levels of CB1 transcripts in the urothelium of patients with DO and lower levels in the detrusor, compared with normal bladders. Radioligand binding revealed CB density of 421 ± 104 fmol/mg protein in normal human bladders. IHC confirmed these findings at the protein level. IF staining demonstrated co-localisation of CB1 with choline acetyltransferase-(ChAT)-positive nerves in the detrusor and co-localisation with PGP9.5 in both urothelium and detrusor. CB2 was co-localised with both ChAT and PGP9.5 in the urothelium and the detrusor. Conclusions: Cannabinoid receptor expression is reduced in the detrusor of patients with DO, which may play a role in the pathophysiology of the disease. Co-localisation of CB receptors with cholinergic nerves may suggest that CB1, being localised on pre- and postsynaptic terminals, could influence neurotransmitter release. Our findings suggest the potential role of cannabinoid agonists in overactive bladder pharmacotherapy

Stiffness distribution of cells and results of migration and invasion test.

<p>(A) Box-and-whisker plots of stiffness of single cells for different cell lines, the percentiles are 10%, 25%, 50%, 75% and 90%, the inset shows the representative force curves of IOSE and HEY. Overall difference among means is significant (p-value<2.2×10⁻¹⁶, Kruskal-Wallis); pairwise differences are significant between IOSE and HEY, HEY A8 and OVCAR-3 cells, between HEY A8 and HEY cells and between HEY A8 and OVCAR-4 cells (p<0.05, Dunn’s post test); (B) Migration and invasion tests for IOSE, HEY and HEY A8 cells. F(480/520) is a fluorescence intensity at 480 nm excitation and 520 nm emission, which is proportional to the number of migrating or invading cells.</p

Cell Stiffness Is a Biomarker of the Metastatic Potential of Ovarian Cancer Cells

<div><p>The metastatic potential of cells is an important parameter in the design of optimal strategies for the personalized treatment of cancer. Using atomic force microscopy (AFM), we show, consistent with previous studies conducted in other types of epithelial cancer, that ovarian cancer cells are generally softer and display lower intrinsic variability in cell stiffness than non-malignant ovarian epithelial cells. A detailed examination of highly invasive ovarian cancer cells (HEY A8) relative to their less invasive parental cells (HEY), demonstrates that deformability is also an accurate biomarker of metastatic potential. Comparative gene expression analyses indicate that the reduced stiffness of highly metastatic HEY A8 cells is associated with actin cytoskeleton remodeling and microscopic examination of actin fiber structure in these cell lines is consistent with this prediction. Our results indicate that cell stiffness may be a useful biomarker to evaluate the relative metastatic potential of ovarian and perhaps other types of cancer cells.</p> </div

2.5D Vehicle odometry estimation

t is well understood that in ADAS applications, a good estimate of the pose of the vehi cle is required. This paper proposes a metaphorically named 2.5D odometry, whereby the planar odometry derived from the yaw rate sensor and four wheel speed sensors is aug mented by a linear model of suspension. While the core of the planar odometry is a yaw rate model that is already understood in the literature, this is augmented by fitting a quadratic to the incoming signals, enabling interpolation, extrapolation, and a finer integration of the vehicle position. It is shown, by experimental results with a DGPS/IMU reference, that this model provides highly accurate odometry estimates, compared with existing methods. Utilising sensors that return the change in height of vehicle reference points with chang ing suspension configurations, a planar model of the vehicle suspension is defined, thus augmenting the odometry model. An experimental framework and evaluations criteria is presented by which the goodness of the odometry is evaluated and compared with existing methods. This odometry model has been designed to support low-speed surround-view camera systems that are well-known. Thus, some application results that show a perfor mance boost for viewing and computer vision applications using the proposed odometry are presented

Significantly enriched Genego Process Networks (p-val: p-value for hypergeometric distribution; ratio: number of mapped genes to total number of genes).

<p>Significantly enriched Genego Process Networks (p-val: p-value for hypergeometric distribution; ratio: number of mapped genes to total number of genes).</p