Molecular expression patterns in the synovium and their association with advanced symptomatic knee osteoarthritis.

OBJECTIVE: Osteoarthritis (OA) is a major source of knee pain. Mechanisms of OA knee pain are incompletely understood but include synovial pathology. We aimed to identify molecular expression patterns in the synovium associated with symptomatic knee OA. DESIGN: Snap frozen synovia were from people undergoing total knee replacement (TKR) for advanced OA, or from post-mortem (PM) cases who had not sought help for knee pain. Associations with OA symptoms were determined using discovery and validation samples, each comprising TKR and post mortem (PM) cases matched for chondropathy (Symptomatic or Asymptomatic Chondropathy). Associations with OA were determined by comparing age matched TKR and PM control cases. Real-time quantitative PCR for 96 genes involved in inflammation and nerve sensitisation used TaqMan® Array Cards in discovery and validation samples, and protein expression for replicated genes was quantified using Luminex bead assay. RESULTS: Eight genes were differentially expressed between asymptomatic and symptomatic chondropathy cases and replicated between discovery and validation samples (P<0.05 or >3-fold change). Of these, matrix metalloprotease (MMP)-1 was also increased whereas interleukin-1 receptor 1 (IL1R1) and vascular endothelial growth factor (VEGF) were decreased at the protein level in the synovium of symptomatic compared to asymptomatic chondropathy cases. MMP1 protein expression was also increased in OA compared to PM controls. CONCLUSION: Associations of symptomatic OA may suggest roles of MMP1 expression and IL1R1 and VEGF pathways in OA pain. Better understanding of which inflammation-associated molecules mediate OA pain should inform refinement of existing therapies and development of new treatments.

The anti-NGF antibody muMab 911 both prevents and reverses pain behaviour and subchondral osteoclast numbers in a rat model of osteoarthritis pain.

OBJECTIVE: Nerve growth factor (NGF) has a pivotal role in peripheral hyperalgesia and inflammation; anti-NGF antibodies attenuate pain responses in inflammatory pain models, and in people with osteoarthritis (OA) or low back pain. The aim of this study was to characterise the peripheral mechanisms contributing to the analgesic effects of anti-NGF antibody treatment in an established model of joint pain, which mimics key clinical features of OA. DESIGN: Effects of preventative vs therapeutic treatment with an anti-NGF antibody (monoclonal antibody 911: muMab 911 (10 mg/kg, s.c.)) on pain behaviour (weight bearing asymmetry and hindpaw withdrawal thresholds (PWT)), cartilage damage, synovitis and numbers of subchondral osteoclasts were investigated in the monosodium iodoacetate (MIA) model. Potential direct effects of NGF on receptor activator of nuclear factor kappa-B ligand (RANKL) mediated osteoclastogenesis were investigated in cultured human osteoclasts. RESULTS: Intra-articular MIA injection resulted in significant pain behaviour, cartilage damage, synovitis and increased numbers of subchondral osteoclasts. Both preventative and therapeutic treatment with muMab 911 significantly prevented, or reversed, MIA-induced pain behaviour, but did not alter cartilage or synovial pathology quantified at the end of the treatment period. NGF did not facilitate RANKL driven osteoclast differentiation in vitro, but preventative or therapeutic muMab 911 reduced numbers of TRAP positive osteoclasts in the subchondral bone. CONCLUSIONS: We demonstrate that anti-NGF antibody treatment attenuates OA pain behaviour despite permitting cartilage damage and synovitis. Indirect effects on subchondral bone remodelling may contribute to the analgesic effects of NGF blockade.

Exome sequencing-driven discovery of coding polymorphisms associated with common metabolic phenotypes.

AIMS/HYPOTHESIS: Human complex metabolic traits are in part regulated by genetic determinants. Here we applied exome sequencing to identify novel associations of coding polymorphisms at minor allele frequencies (MAFs) >1% with common metabolic phenotypes. METHODS: The study comprised three stages. We performed medium-depth (8×) whole exome sequencing in 1,000 cases with type 2 diabetes, BMI >27.5 kg/m(2) and hypertension and in 1,000 controls (stage 1). We selected 16,192 polymorphisms nominally associated (p < 0.05) with case-control status, from four selected annotation categories or from loci reported to associate with metabolic traits. These variants were genotyped in 15,989 Danes to search for association with 12 metabolic phenotypes (stage 2). In stage 3, polymorphisms showing potential associations were genotyped in a further 63,896 Europeans. RESULTS: Exome sequencing identified 70,182 polymorphisms with MAF >1%. In stage 2 we identified 51 potential associations with one or more of eight metabolic phenotypes covered by 45 unique polymorphisms. In meta-analyses of stage 2 and stage 3 results, we demonstrated robust associations for coding polymorphisms in CD300LG (fasting HDL-cholesterol: MAF 3.5%, p = 8.5 × 10(-14)), COBLL1 (type 2 diabetes: MAF 12.5%, OR 0.88, p = 1.2 × 10(-11)) and MACF1 (type 2 diabetes: MAF 23.4%, OR 1.10, p = 8.2 × 10(-10)). CONCLUSIONS/INTERPRETATION: We applied exome sequencing as a basis for finding genetic determinants of metabolic traits and show the existence of low-frequency and common coding polymorphisms with impact on common metabolic traits. Based on our study, coding polymorphisms with MAF above 1% do not seem to have particularly high effect sizes on the measured metabolic traits.

Lightning-induced extensive charge sheets provide long range electrostatic thunderstorm detection.

By combining electrostatic measurements of lightning-induced electrostatic field changes with radio frequency lightning location, some field changes from exceptionally distant lightning events are apparent which are inconsistent with the usual inverse cube of distance. Furthermore, by using two measurement sites, a transition zone can be identified beyond which the electric field response reverses polarity. For these severe lightning events, we infer a horizontally extensive charge sheet above a thunderstorm, consistent with a mesospheric halo of several hundred kilometers' extent.

Electrical control of the exciton fine structure of a quantum dot molecule.

We electrically control the coherent mixing of the optically bright spin states of excitons confined in InAs/GaAs quantum dot molecules. By tunnel coupling two quantum dots, using a vertical electric field, the exciton fine structure splitting and eigenstate orientation relative to the crystal lattice are tuned. We model the electric field dependent anisotropic electron-hole exchange interaction accurately and propose that the controllable mixing of the spin states will enable electrically controlled quantum operations on exciton spin qubits.

Indistinguishable entangled photons generated by a light-emitting diode.

A linear optical quantum computer relies on interference between photonic qubits for logic, and entanglement for near-deterministic operation. Here we measure the interference and entanglement properties of photons emitted by a quantum dot embedded within a light-emitting diode. We show that pairs of simultaneously generated photons are entangled, and indistinguishable from subsequently generated photons. We measure entanglement fidelity of 0.87 and two-photon-interference visibility of 0.60 ± 0.05. The visibility, limited by detector jitter, could be improved by optical cavity designs.

Challenges to bonnet monkey (Macaca radiata) social groups: Mother-infant dyad and infant social interactions.

The mother-infant dyad is crucial to early development in a variety of species. The complexity of social groupings in nonhuman primates makes this relationship resilient as well as susceptible to early challenges associated with environmental chaos. Quantitative behavior observations of bonnet monkey mother-infant interactions were collected from 28 mother-infant dyads between one and twelve months of age. Social groups were subjected to several prenatal and/or postnatal housing relocations within a single year resulting in two study groups. One group experienced relocations (ATYPICAL, n = 14) and the second group (TYPICAL, n = 14) was conceived and reared in the same location. Behaviors in the ethogram included mother-infant interactions and infant social interactions with other members of the group. Observations between ages of two to four months were analyzed by a mixed model analysis of variance including fixed effects of per and postnatal history (TYPICAL, ATYPICAL), age, and history by age interaction and random effects of mother and infant nested within mother. A significant effect of relocation history was noted on a number of infant behaviors. ATYPICAL infants were out of direct contact with their mother at an earlier age but remained in her proximity. Control of proximity shifted to offsrping in the ATYPICAL group compared to the TYPICAL group. Furthermore, greater social interactions between two and four months of age with other members of the social group as well as the ir mother were observed in the ATYPICAL group. It is suggested that continuous challenge associated with relocation may affect the infant at later developmental ages due to these early differences in ways that are yet unclear.

Cannabinoid activation of PPAR alpha; a novel neuroprotective mechanism.

BACKGROUND AND PURPOSE: Although CB(1) receptor activation evokes neuroprotection in response to cannabinoids, some cannabinoids have been reported to be peroxisome proliferator activated receptor (PPAR) ligands, offering an alternative protective mechanism. We have, therefore, investigated the ability of a range of cannabinoids to activate PPAR alpha and for N-oleoylethanolamine (OEA), an endogenous cannabinoid-like compound (ECL), to evoke neuroprotection. EXPERIMENTAL APPROACH: Assays of PPAR alpha occupancy and gene transactivation potential were conducted in cell-free and transfected HeLa cell preparations, respectively. In vivo estimates of PPAR alpha activation through fat mobilization and gene transcription were conducted in mice. Neuroprotection in vivo was investigated in wild-type and PPAR alpha gene-disrupted mice. KEY RESULTS: The ECLs OEA, anandamide, noladin ether and virodhamine were found to bind to the purified PPAR alpha ligand binding domain and to increase PPAR alpha-driven transcriptional activity. The high affinity synthetic CB(1/2) cannabinoid agonist WIN 55212-2 bound to PPAR alpha equipotently with the PPARalpha agonist fenofibrate, and stimulated PPARalpha-mediated gene transcription. The phytocannabinoid delta 9 tetrahydrocannabinol was without effect. OEA and WIN 55212-2 induced lipolysis in vivo, while OEA pre-treatment reduced infarct volume from middle cerebral artery occlusion in wild-type, but not in PPAR alpha-null mice. OEA treatment also led to increased expression of the NFkappa B-inhibitory protein, Ikappa B, in mouse cerebral cortex, while expression of the NFkappa B-regulated protein COX-2 was inhibited. CONCLUSIONS AND IMPLICATIONS: These data demonstrate the potential for a range of cannabinoid compounds, of diverse structures, to activate PPAR alpha and suggest that at least some of the neuroprotective properties of these agents could be mediated by nuclear receptor activation.

Effects of the March 2015 solar eclipse on near-surface atmospheric electricity.

Measurements of atmospheric electrical and standard meteorological parameters were made at coastal and inland sites in southern England during the 20 March 2015 partial solar eclipse. Clear evidence of a reduction in air temperature resulting from the eclipse was found at both locations, despite one of them being overcast during the entire eclipse. The reduction in temperature was expected to affect the near-surface electric field (potential gradient (PG)) through a reduction in turbulent transfer of space charge. No such effect could be unambiguously confirmed, however, with variability in PG and air-Earth current during the eclipse being comparable to pre- and post-eclipse conditions. The already low solar radiation for this latitude, season and time of day was likely to have contributed to the reduced effect of the eclipse on atmospheric electricity through boundary layer stability. The absence of a reduction in mean PG shortly after time of maximum solar obscuration, as observed during eclipses at lower geomagnetic latitude, implied that there was no significant change in atmospheric ionization from cosmic rays above background variability. This finding was suggested to be due to the relative importance of cosmic rays of solar and galactic origin at geomagnetic mid-latitudes.This article is part of the themed issue 'Atmospheric effects of solar eclipses stimulated by the 2015 UK eclipse'.

A semiconductor photon-sorter.

Obtaining substantial nonlinear effects at the single-photon level is a considerable challenge that holds great potential for quantum optical measurements and information processing. Of the progress that has been made in recent years one of the most promising methods is to scatter coherent light from quantum emitters, imprinting quantum correlations onto the photons. We report effective interactions between photons, controlled by a single semiconductor quantum dot that is weakly coupled to a monolithic cavity. We show that the nonlinearity of a transition modifies the counting statistics of a Poissonian beam, sorting the photons in number. This is used to create strong correlations between detection events and to create polarization-correlated photons from an uncorrelated stream using a single spin. These results pave the way for semiconductor optical switches operated by single quanta of light.

Disclosure and prevention of medical errors. Committee on Bioethical Issues of the Medical Society of the State of New York.

Medical errors occur and are sometimes unavoidable. Physicians generally, but not always, have ethical and moral obligations to disclose their errors to the patient. Because common medical errors can be expected, physicians are obligated to work within health systems toward reducing systems flaws that promote errors. However, the obligations of physicians to disclose errors made by others are less clear. This article discusses the professional ethics involved in disclosing and preventing medical errors.

No evidence for linkage at candidate type 2 diabetes susceptibility loci on chromosomes 12 and 20 in United Kingdom Caucasians.

Several studies have identified evidence for linkage between type 2 diabetes and the regions on chromosomes 12 and 20 containing the maturity-onset diabetes of the young (MODY) genes, hepatocyte nuclear factor-1alpha (HNF-1alpha) and HNF-4alpha. Two studies examining the HNF-1alpha region have demonstrated evidence for linkage at genome-wide levels of significance, whereas four studies examining the HNF-4alpha locus have resulted in evidence for linkage at more suggestive levels of significance. The demonstration of linkage to these regions in additional patient series will strengthen the evidence that susceptibility alleles exist at these loci. We therefore assessed the evidence for linkage to these regions using a large cohort of United Kingdom Caucasian type 2 diabetes-affected sibling pairs. A maximum total of 315 affected full sibling pairs were typed for microsatellite markers across the MODY regions and, in a subset of families, for markers spanning the whole of chromosome 20. Evidence for linkage was assessed using a multipoint, mode of inheritance-free method. Linkage analysis did not reveal any significant evidence for excess allele sharing at any of the regions studied. Loci contributing sibling recurrence risks, relative to the general population risk, of 1.75 and 1.25 could be excluded for the HNF-1alpha and HNF-4alpha regions, respectively. We have not confirmed in United Kingdom Caucasians the evidence for linkage previously reported on 12q and 20q. Our results highlight further the problems of replicating previous positive linkage results across different ethnic groups.

Polymorphic debrisoquine 4-hydroxylase activity in the rat is due to differences in CYP2D2 expression.

The female Dark Agouti rat is widely used as an animal model for the CYP2D6 poor metabolizer phenotype, males of other strains such as Sprague Dawley or Wistar serving as models for the extensive metabolizer phenotype. To determine the relative level of expression of CYP2D enzymes in the liver of female and male Dark Agouti, Sprague Dawley and Wistar rats, anti-peptide antibodies were raised in rabbits against short synthetic peptides representing the C-termini of the rat P450 enzymes CYP2D1, CYP2D2, CYP2D3, CYP2D4 and CYP2D5. In immunoblotting studies, it was found that the hepatic expression of CYP2D1 was greater in Dark Agouti rats than Sprague Dawley or Wistar rats. In contrast, hepatic CYP2D2 was 30-40-fold less abundant in female Dark Agouti than female Sprague Dawley or Wistar rats and six- to eightfold less abundant in male Dark Agouti than male Sprague Dawley or Wistar rats. No hepatic CYP2D3 could be detected in either sex of any of the three strains. Hepatic CYP2D4 expression was generally greater in male than female rats, and higher in Dark Agouti compared with Sprague Dawley or Wistar strains. CYP2D5 was expressed in the livers of female and male Dark Agouti rats but not in female Sprague Dawley or Wistar rats. This form was variably expressed in livers of male Sprague Dawley and Wistar rats. Hepatic debrisoquine 4-hydroxylase activity was markedly reduced in female and male Dark Agouti rats as compared to Sprague Dawley or Wistar rats and correlated (r = 0.88; P < 0.001) with the hepatic CYP2D2 content. Recombinant CYP2D2 was 18-fold more active at catalysing the 4-hydroxylation of debrisoquine than CYP2D1. Furthermore, quinine markedly inhibited CYP2D2-mediated debrisoquine and metoprolol oxidation, while quinidine, its diastereoisomer, inhibited the reactions to a lesser extent. In conclusion, these results show that impaired debrisoquine 4-hydroxylase activity in the female Dark Agouti rat is due to low levels of CYP2D2.

Hepatic microsomal triglyceride transfer protein messenger RNA concentrations are increased by dietary cholesterol in hamsters.

In hamsters fed high fat diets enriched in trimyristin, tripalmitin or tristearin, increased dietary cholesterol content was associated with increased plasma concentrations of very low density lipoprotein (VLDL) cholesterol and triacylglycerol (p < 0.0001 and p = 0.0017, respectively). Hepatic microsomal triglyceride transfer protein (MTP) mRNA concentration also increased (p < 0.0001), independent of the nature of dietary fat, and was significantly correlated with the plasma VLDL lipid concentrations (p = 0.0002 and p = 0.0106 for cholesterol and triacylglycerol, respectively) and hepatic cholesterol concentrations. Increased expression of the MTP gene may be part of a coordinated response to hepatic cholesterol accumulation leading to increased VLDL lipid secretion.

Local kallikrein and trypsin responses in the rat.

1 Locally administered commercial hog pancreatic kallikrein (Depot-Glumorin) and bovine pancreatic trypsin both increased vascular permeability in the skin and paws of rats. 2 By the use of numerous antagonists and enzyme inhibitors, this vascular response was found to be the result not of kinin formation but of a direct action mostly on histamine receptors. 3 Highly purified kallikrein did not increase vascular permeability in rats, suggesting either that the effect was due to an impurity in the commercial preparation or that a structural change in the enzyme occurred on purification. 4 Soya bean trypsin inhibitor prevented the trypsin response when both were injected locally. On intraperitoneal injection, the inhibitor was effective only against local kallikrein. 5 The kallikrein inhibitor, aprotinin (Trasylol), was not effective against local kallikrein but it reduced the trypsin response when both were injected locally.

Molecular cloning and nucleotide sequence analysis of the maltose-inducible porin gene of Aeromonas salmonicida.

The gene for the Aeromonas salmonicida maltose-inducible porin (maltoporin) was cloned into phagemid pTZ18R in two restriction fragments, 0.6-kb PstI/KpnI and 1.7-kb SphI, of genomic DNA and their nucleotide sequences were determined. Open reading frames of 1329 and 1335 bp translated into sequences of 443 and 445 amino acids, with a 23 or 25 amino acid signal sequence and a 420 amino acid mature protein of molecular mass 46424 Da. Putative ribosome binding sites, AGGA and GGGAA, occurred 9 bp upstream of two possible ATG initiation codons. The A. salmonicida gene product showed a high degree of similarity with Escherichia coli LamB, and codon usage was very similar to that of another A. salmonicida outer membrane protein but markedly different from those of extracellular proteins.

Genetic determinants of plasma lipoprotein levels and their dietary response.

Hamsters were fed diets containing fat (5-20%, w/w) consisting of triolein (TO) alone or 50% triolein plus 50% trimyristin (TM), tripalmitin (TP) or tristearin (TS) for 28 days. Each fat had unique effects on lipoprotein concentrations which were related to changes in the expression of hepatic genes. Tripalmitin was the most hypercholesterolaemic of the saturated fats, causing dose-dependent increases in LDL and HDL cholesterol which correlated with decreases in the expression of HMGCoA reductase and LDL receptor genes. Tripalmitin also increased the expression of the apoB gene. It seems likely that fatty acids may regulate genes which are involved both in the synthesis and clearance of plasma lipoproteins. Inclusion of increasing amounts of cholesterol in diets containing 20% fat (50% TO plus 50% TP or TS) caused down-regulation of HMGCoAR and LDLR genes and up-regulation of the microsomal triglyceride transfer protein (MTP) gene for both fats. This was accompanied by increased LDL-cholesterol in the TP but not the TS group. In all experiments the concentration of VLDL-cholesterol correlated with the hepatic cholesterylester and MTP mRNA concentrations.

Ondansetron modulates pharmacodynamic effects of ketamine on electrocardiographic signals in rhesus monkeys.

Electrocardiographic signal dynamics were examined in rhesus monkeys (Macaca mulatta) before and after treatment with ketamine and/or ondansetron. Ketamine exerts differential pharmacodynamic effects on behavior in animals stratified according to a measure of central serotonergic turnover. We hypothesized that measures of serotonergic turnover might explain some of the variance in the electrocardiographic (ECG) response to ketamine. Electrocardiographic recordings of animals were obtained at baseline, after administration of either saline or ondansetron (0.125 mg/kg), and after administration of ketamine (15 mg/kg). Electrocardiographic signal dynamics were measured using an algorithm that extracts the Hurst parameter (H) of the interbeat interval (IBI) time-series. H decreased after ketamine administration, (mean+/-S.E.M.), 0.33+/-0.04 vs. 0.12+/-0.02, P

Initial ethanol exposure results in decreased heart rate variability in ethanol-naive rhesus monkeys.

Ethanol's effects on heart rate variability may contribute to the increased cardiac disease and mortality observed in alcoholics. We assessed cardiac response to ethanol in seven previously ethanol-naive monkeys given a standard dose of ethanol, or saline. Ethanol exposure reduced cardiac signal complexity [mean+/-S.D. (ethanol: Hurst parameter=0.39+/-0.02; saline: Hurst parameter=0.32+/-0.06)] and increased the spectral exponent (ethanol: beta=1.36+/-0.35; saline: beta=1.12+/-0.35) when compared to saline, while heart rate itself was unaffected (saline: interbeat interval=303.57+/-24.57; ethanol: interbeat interval=308.14+/-20.45). Taken together with data that show autonomic disregulation in alcoholics, these findings provide further evidence of deleterious ethanol effects on cardiac signal dynamics.

Handedness and approach-avoidance behavior in chimpanzees (Pan).

The relationship between hand preference and approach-avoidance behavior was examined in 49 chimpanzees (Pan). Ss were presented with 2 sets of novel objects on 4 consecutive days. The objects were presented for 2 hr during each session, and latency to touch any object was recorded for each S. Latency scores were then compared for chimpanzees that had been determined to be non-right- or right-handed. Right-handed Ss approached and touched the objects significantly faster than non-right-handed Ss did. In addition, males touched the objects significantly faster than did females. Correlations in approach-avoidance behavior were significant across stimulus sets and days of testing. The overall results support recent theoretical models linking hemispheric specialization with the expression of positive and negative affective behaviors.