In vitro sepsis upregulates Nociceptin/Orphanin FQ receptor expression and function on human T- but not B-cells.

  Background and Purpose In animal models of sepsis, increased activation of the Nociceptin/Orphanin FQ (N/OFQ) receptor NOP is associated with mortality and NOP antagonists improved survival. We have explored the role of the N/OFQ-NOP system in freshly isolated volunteer human B- and T-cells incubated with lipopolysaccharide (LPS) and peptidoglycan G (PepG) as a model of in vitro sepsis. Experimental Approach B- and T-cell NOP expression was measured using the NOP fluorescent probe N/OFQATTO594, N/OFQ content was measured using immunofluorescence, N/OFQ release was tracked using a CHOhNOPGαiq5 biosensor assay, NOP function was measured using transwell migration and cytokine/chemokine release using a 25plex assay format. Cells were challenged with LPS/PepG. Key Results CD19 positive B-cells bound N/OFQATTO594; they also contain N/OFQ. Stimulation with CXCL13/IL-4 increased N/OFQ release. N/OFQ trended to reduced migration to CXCL13/IL4. Surface expression was unaffected by LPS/PepG but this treatment increased GM-CSF release in an N/OFQ sensitive manner. CD3 positive T-cells did not bind N/OFQATTO594; they did contain N/OFQ. Stimulation with CXCL12/IL-6 increased N/OFQ release. When incubated with LPS/PepG NOP surface expression was induced leading to N/OFQATTO594 binding. In LPS/PepG treated cells N/OFQ reduced migration to CXCL12/IL6. LPS/PepG increased GM-CSF release in an N/OFQ sensitive manner. Conclusion and Implications We suggest both a constitutive and sepsis-inducible N/OFQ-NOP receptor autocrine regulation of B- and T-cell function respectively. These NOP receptors variably inhibit migration and reduce GM-CSF release. These data provide mechanistic insights to the detrimental role for increased N/OFQ signalling in sepsis and suggest a potential role for NOP antagonists as treatments.</p

Preclinical Discovery and Development of oliceridine (Olinvyk®) for the Treatment of Post-Operative Pain

IntroductionOpioids acting at the MOP(mu:µ) receptor produce analgesia but also side-effects. There is debate suggesting opioid receptors produce analgesia via G-protein and side-effects via β-arrestin-2 pathways. Opioids targeting G-proteins over the arrestins (bias) offer potential therapeutic advantages. Oliceridine is a putative MOP, G-protein biased agonist.Areas CoveredOliceridine is selective for MOP receptors with greater activity at G-proteins over arrestins. A substantial body of evidence now points to a simpler pharmacological descriptor of partial agonist. Pre-clinical in vivo data indicates a robust antinociceptive response of shorter duration than morphine. Apollo trials (Phase-III RCT-bunionectomy/abdominoplasty) describe good analgesic efficacy that was non-inferior to morphine with good tolerability and side-effect profile. There is evidence for improved respiratory safety profile. Oliceridine is approved by the FDA.Expert OpinionOliceridine will be an important addition to the clinical armamentarium for use for the management of acute pain severe enough to require an intravenous opioid analgesic and for whom alternative treatments are inadequate. Respiratory advantage and the possibility of reduced abuse potential are possible advantages over the use of traditional opioids. Based on a number of excellent, highly detailed studies, oliceridine should be described as a partial agonist; this ‘label’ does not matter.</div

Colonial lives across the British Empire: imperial careering in the long nineteenth century

No description supplie

Quantifying Interactions of Nucleobase Atoms with Model Compounds for the Peptide Backbone and Glutamine and Asparagine Side Chains in Water

Alkylureas display hydrocarbon and amide groups, the primary functional groups of proteins. To obtain the thermodynamic information that is needed to analyze interactions of amides and proteins with nucleobases and nucleic acids, we quantify preferential interactions of alkylureas with nucleobases differing in the amount and composition of water-accessible surface area (ASA) by solubility assays. Using an established additive ASA-based analysis, we interpret these thermodynamic results to determine interactions of each alkylurea with five types of nucleobase unified atoms (carbonyl sp²O, amino sp³N, ring sp²N, methyl sp³C, and ring sp²C). All alkylureas interact favorably with nucleobase sp²C and sp³C atoms; these interactions become more favorable with an increasing level of alkylation of urea. Interactions with nucleobase sp²O are most favorable for urea, less favorable for methylurea and ethylurea, and unfavorable for dialkylated ureas. Contributions to overall alkylurea–nucleobase interactions from interactions with each nucleobase atom type are proportional to the ASA of that atom type with proportionality constant (interaction strength) α, as observed previously for urea. Trends in α-values for interactions of alkylureas with nucleobase atom types parallel those for corresponding amide compound atom types, offset because nucleobase α-values are more favorable. Comparisons between ethylated and methylated ureas show interactions of amide compound sp³C with nucleobase sp²C, sp³C, sp²N, and sp³N atoms are favorable while amide sp³C–nucleobase sp²O interactions are unfavorable. Strongly favorable interactions of urea with nucleobase sp²O but weakly favorable interactions with nucleobase sp³N indicate that amide sp²N–nucleobase sp²O and nucleobase sp³N–amide sp²O hydrogen bonding (NH···OC) interactions are favorable while amide sp²N–nucleobase sp³N interactions are unfavorable. These favorable amide–nucleobase hydrogen bonding interactions are prevalent in specific protein–nucleotide complexes

MOESM1 of Limb patterning genes and heterochronic development of the emu wing bud

Additional file 1. Cell counting using ImageJ (Fiji). Stage 21 emu forelimb bud is shown for example

Experimental Atom-by-Atom Dissection of Amide–Amide and Amide–Hydrocarbon Interactions in H₂O

Quantitative information about amide interactions in water is needed to understand their contributions to protein folding and amide effects on aqueous processes and to compare with computer simulations. Here we quantify interactions of urea, alkylated ureas, and other amides by osmometry and amide–aromatic hydrocarbon interactions by solubility. Analysis of these data yields strengths of interaction of ureas and naphthalene with amide sp²O, amide sp²N, aliphatic sp³C, and amide and aromatic sp²C unified atoms in water. Interactions of amide sp²O with urea and naphthalene are favorable, while amide sp²O–alkylurea interactions are unfavorable, becoming more unfavorable with increasing alkylation. Hence, amide sp²O–amide sp²N interactions (proposed n−σ* hydrogen bond) and amide sp²O–aromatic sp²C (proposed n−π*) interactions are favorable in water, while amide sp²O–sp³C interactions are unfavorable. Interactions of all ureas with sp³C and amide sp²N are favorable and increase in strength with increasing alkylation, indicating favorable sp³C–amide sp²N and sp³C–sp³C interactions. Naphthalene results show that aromatic sp²C–amide sp²N interactions in water are unfavorable while sp²C–sp³C interactions are favorable. These results allow interactions of amide and hydrocarbon moieties and effects of urea and alkylureas on aqueous processes to be predicted or interpreted in terms of structural information. We predict strengths of favorable urea–benzene and <i>N</i>-methylacetamide interactions from experimental information to compare with simulations and indicate how amounts of hydrocarbon and amide surfaces buried in protein folding and other biopolymer processes and transition states can be determined from analysis of urea and diethylurea effects on equilibrium and rate constants

Nuclear_sequence_alignments

Non-partitioned multiple sequence alignments for various nuclear loci in FASTA format (.fasta)

MCMCTree_inputs

Inputs in phylip format (.phy) for molecular dating analysis conducted in MCMCTree for several datasets (mt=mitochondrial, nuc=nuclear, nt=standard coded (nucleotide), ry=ry-coded, m3=mitochondrial third codon positions). Note that RY-coded data are represented by R and Y. Tree topologies (.trees) used as input are shown: for mitochondrial and combined mitochondrial/nuclear datasets, two topologies were tested, one in which moa/tinamous are sister to the remaining notopalaeognathae, and one where rheas are sister to the remaining notopalaeognathae. For the nuclear dataset, the topology derived from phylogenetic inference on the nuclear data alone was used. MCMCTree control files (.ctl) are not shown as parameters used are described in text

MrBayes_inputs

Input nexus files (.nex) used for Bayesian phylogenetic inference for various datasets (mt=mitochondrial, nuc=nuclear, nt=standard coded (nucleotide), ry=ry-coded, m3=mitochondrial third codon positions, loops=mitochondrial rRNA loops

RaxML_inputs

Input phylip files (.phy) and partition files (_part.txt) used for maximum likelihood tree generation in RaxML for various data sets (mt=mitochondrial, nuc=nuclear, ry=ry-coded, m3=mitochondrial third codon positions, loops=mitochondrial rRNA loops). Note that RY coded bases are given by a 0 (R) or 1 (Y)