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Genetic variations in the human mu-opioid receptor gene (OPRM1) mediate individual differences in response to pain and opiate addiction. We studied whether the common A118G (rs1799971) mu-opioid receptor single nucleotide polymorphism (SNP) was associated with overdose severity in humans. In addition, we examined an SNP responsible for alternative splicing of OPRM1 (rs2075572). We assessed allele frequencies of the above SNPs and associations with clinical severity in patients presenting to the emergency department (ED) with acute drug overdose. This work was designed as an observational cohort study over a 12-month period at an urban teaching hospital. Participants consisted of consecutive adult ED patients with suspected acute drug overdose for whom discarded blood samples were available for analysis. Specimens were linked with clinical variables (demographics, urine toxicology screens, clinical outcomes) then deidentified prior to genetic SNP analysis. Blinded genotyping was performed after standard DNA purification and whole genome amplification. In-hospital severe outcomes were defined as either respiratory arrest (RA; defined by mechanical ventilation) or cardiac arrest (CA; defined by loss of pulse). We analyzed 179 patients (61% male, median age 32) who overall suffered 15 RAs and four CAs, of whom three died. The 118G allele conferred 5.3-fold increased odds of CA/RA (p

Activation of mu opioid receptors within the ventral tegmental area (VTA) can produce reward through the inhibition of GABAergic inputs. GABAergic neurons in the ventral pallidum (VP) provide a major input to VTA neurons. To determine the specific VTA neuronal targets of VP afferents and their sensitivity to mu opioid receptor agonists, we virally expressed channel rhodopsin (ChR2) in rat VP neurons and optogenetically activated their terminals in the VTA. Light activation of VP neuron terminals elicited GABAergic IPSCs in both dopamine (DA) and non-DA VTA neurons, and these IPSCs were inhibited by the mu opioid receptor agonist DAMGO. In addition, using a fluorescent retrograde marker to identify VTA-projecting VP neurons, we found them to be hyperpolarized by DAMGO. Both of these actions decrease GABAergic input onto VTA neurons, revealing two mechanisms by which endogenous or exogenous opioids can activate VTA neurons, including DA neurons.

Morphine and related µ-opioid receptor (MOR) agonists remain among the most effective drugs known for acute relief of severe pain. A major problem in treating painful conditions is that tolerance limits the long-term utility of opioid agonists. Considerable effort has been expended on developing an understanding of the molecular and cellular processes that underlie acute MOR signaling, short-term receptor regulation, and the progression of events that lead to tolerance for different MOR agonists. Although great progress has been made in the past decade, many points of contention and controversy cloud the realization of this progress. This review attempts to clarify some confusion by clearly defining terms, such as desensitization and tolerance, and addressing optimal pharmacological analyses for discerning relative importance of these cellular mechanisms. Cellular and molecular mechanisms regulating MOR function by phosphorylation relative to receptor desensitization and endocytosis are comprehensively reviewed, with an emphasis on agonist-biased regulation and areas where knowledge is lacking or controversial. The implications of these mechanisms for understanding the substantial contribution of MOR signaling to opioid tolerance are then considered in detail. While some functional MOR regulatory mechanisms contributing to tolerance are clearly understood, there are large gaps in understanding the molecular processes responsible for loss of MOR function after chronic exposure to opioids. Further elucidation of the cellular mechanisms that are regulated by opioids will be necessary for the successful development of MOR-based approaches to new pain therapeutics that limit the development of tolerance.

The mu and delta types of opioid receptors form heteromers that exhibit pharmacological and functional properties distinct from those of homomeric receptors. To characterize these complexes in the brain, we generated antibodies that selectively recognize the mu-delta heteromer and blocked its in vitro signaling. With these antibodies, we showed that chronic, but not acute, morphine treatment caused an increase in the abundance of mu-delta heteromers in key areas of the central nervous system that are implicated in pain processing. Because of its distinct signaling properties, the mu-delta heteromer could be a therapeutic target in the treatment of chronic pain and addiction.

Electrode arrays are sometimes implanted in the brains of patients with intractable epilepsy to better localize seizure foci before epilepsy surgery. Analysis of intracranial EEG (iEEG) recordings is typically performed in the electrode channel domain without explicit separation of the sources that generate the signals. However, intracranial EEG signals, like scalp EEG signals, could be linear mixtures of local activity and volume-conducted activity arising in multiple source areas. Independent component analysis (ICA) has recently been applied to scalp EEG data, and shown to separate the signal mixtures into independently generated brain and non-brain source signals. Here, we applied ICA to unmix source signals from intracranial EEG recordings from four epilepsy patients during a visually cued finger movement task in the presence of background pathological brain activity. This ICA decomposition demonstrated that the iEEG recordings were not maximally independent, but rather are linear mixtures of activity from multiple sources. Many of the independent component (IC) projections to the iEEG recording grid were consistent with sources from single brain regions, including components exhibiting classic movement-related dynamics. Notably, the largest IC projection to each channel accounted for no more than 20-80% of the channel signal variance, implying that in general intracranial recordings cannot be accurately interpreted as recordings of independent brain sources. These results suggest that ICA can be used to identify and monitor major field sources of local and distributed functional networks generating iEEG data. ICA decomposition methods are useful for improving the fidelity of source signals of interest, likely including distinguishing the sources of pathological brain activity.

The application of the metamaterial (MM) in wireless power transfer (WPT) system has received a great deal of attention in improving the transfer efficiency and reducing the leakage of electromagnetic field (EMF) in recent years. However, most of the analytical methods for MM‐inspired WPT system are based on microwave technologies, such as negative refraction effect, magnetic dipole coupling model and magneto‐inductive wave theory. In this study, the equivalent circuit model of the MM in the WPT system is analysed in detail. Two different types of MMs, Mu‐Negative‐Magnetic (MNM) and Mu‐Near‐Zero (MNZ), are constructed to verify this method. In particular, a tuning strategy by changing the distances between the drive (load) loop and internal coils is established to improve the power transfer efficiency for the WPT system with the MNM‐MM. In addition, the MNZ‐MM is described to cancel EMF leakage in the WPT systems by this method. Moreover, a series of practical experiments are carried out to prove the numerical analysis, which are well consistent with the theoretical solution employed by the Advanced Design System software. Finally, the combination of two types of MMs applied in the WPT system is also discussed. [ABSTRACT FROM AUTHOR]

Development of bidirectional non-monotonic segmented leaf sequence (NSLS) MLC delivery technique compatible with Varian MLC for non-split IMRT fields reducing total monitor units (TotalMU) and the number of segments (NS) simultaneously and assessment of its efficiency using a plan scoring index (PSI). The optimal fluence of IMRT plans of ten patients of lung carcinoma, calculated using Eclipse TPS version 11.0 (Varian Medical Systems, Palo Alto, CA, USA), was used to generate the segmented MLC fields using our newly developed equally spaced (ES) reducing level and NSLS algorithms in MATLAB® version 2011b for 6–10 intensity levels. These MLC fields were imported into the plans with the same field setup and the final dose was recalculated. The results were compared with those of commercially available multiple static segments (MSS) leaf motion calculation (LMC) algorithm and few previously published algorithms. Plan scoring index (PSI) and degree of modulation (DoM) was calculated to compare the quality of different plans for the same patient. The average differences in TotalMU and NS with respect to MSS algorithm are −3.80% and −14.28% for the NSLS algorithm, respectively. The calculated average PSI and DoM is 0.75, 2.51 and 0.91, 2.41 for the MSS and NSLS algorithms, respectively. IMRT plans generated using the NSLS algorithm resulted in the best PSI, DoM values among all the leaf sequencing algorithms. Our proposed NSLS algorithm allows bidirectional delivery in Varian medical linear accelerator which is not commercially available. NSLS algorithm is efficient in reducing the TotalMU and NS with equivalent plan quality as that of MSS. [ABSTRACT FROM AUTHOR]

• Pairs of 10 Hz oscillations from homologous left and right cortices show a small frequency separation. • This frequency dissociation may help homologous cortices avoid obligatory synchronization. • Prior theory related frequency separation with metastable brain dynamics, increased connectional flexibility. Using high resolution spectral methods to uncover neuromarkers of social, cognitive and behavioral function, we have found that hemi-lateralized pairs of oscillations such as left and right occipital alpha or left and right rolandic mu dissociate spectrally. That is, they show a shifted frequency distribution, with one member of the pair peaking at a slightly lower frequency than the other. Resorting to the analysis of EEG spatio-spectral patterns, we provide examples of dissociations in the 10Hz frequency band. Occasionally, hemi-lateralized pairs blend into medial aggregates, probably when functional interactions lead to strongly coherent dynamics through frequency-locking or metastability. Our observations support the hypothesis that homologous pairs of neuromarkers have characteristically distinct intrinsic frequencies and coordinate their oscillations into synchronous ensembles only transiently. This property could play a role in the balance of integration and segregation in the brain: spectral separation of the oscillations from homologous cortical areas allows them to function independently under certain circumstances, all the while preserving a potential for stronger interactions supported by structural and functional symmetries. Spectral dissociation (and its methodological corollary: spectral analysis with high frequency resolution) may be harnessed to better track the individual power of each member of a hemi-lateralized pair and their respective time-course, leading to enhanced internal validity and reproducibility of research on neural oscillations. Resulting insights may shed light on the functional interaction between homologous cortices in studies of attention (alpha), e.g. during perceptual and social interaction tasks, and in studies of somatomotor processing (mu), e.g. in bimanual coordination and neuroprosthetics. [ABSTRACT FROM AUTHOR]

This article analyses myths regarding so-called "dwarf people," which were recorded in the region of Polynesia. According to many of these myths, these people were ancient ancestors of currentday Polynesians. Common characteristics of these people across various myths include their very short stocky stature, living in deep valleys and forests hidden away from others. All myths about Menehune highlight their exceptional skill in working with stone and building various structures such as shrines, waterways, ponds, roads, and others. Much Hawaiian mythology attributes many stone structures to their work. Anthropologists and scholars of religion favour the opinion that these "dwarf people" were purely mythical beings. However, since the discovery of skeletons of small people on Flores Island in 2003, this topic can now be viewed from a completely different angle. [ABSTRACT FROM AUTHOR]

Electroencephalography (EEG) activity in the mu frequency band (8–13 Hz) is suppressed during both gesture performance and observation. However, it is not clear if or how particular characteristics within the kinematic execution of gestures map onto dynamic changes in mu activity. Mapping the time course of gesture kinematics onto that of mu activity could help understand which aspects of gestures capture attention and aid in the classification of communicative intent. In this work, we test whether the timing of inflection points within gesture kinematics predicts the occurrence of oscillatory mu activity during passive gesture observation. The timing for salient features of performed gestures in video stimuli was determined by isolating inflection points in the hands' motion trajectories. Participants passively viewed the gesture videos while continuous EEG data was collected. We used wavelet analysis to extract mu oscillations at 11 Hz and at central electrodes and occipital electrodes. We used linear regression to test for associations between the timing of inflection points in motion trajectories and mu oscillations that generalized across gesture stimuli. Separately, we also tested whether inflection point occurrences evoked mu/alpha responses that generalized across participants. Across all gestures and inflection points, and pooled across participants, peaks in 11 Hz EEG waveforms were detected 465 and 535 ms after inflection points at occipital and central electrodes, respectively. A regression model showed that inflection points in the motion trajectories strongly predicted subsequent mu oscillations ( R 2 = 0.921 , p <0.01); effects were weaker and non-significant for low (17 Hz) and high (21 Hz) beta activity. When segmented by inflection point occurrence rather than stimulus onset and testing participants as a random effect, inflection points evoked mu and beta activity from 308 to 364 ms at central electrodes, and broad activity from 226 to 800 ms at occipital electrodes. The results suggest that inflection points in gesture trajectories elicit coordinated activity in the visual and motor cortices, with prominent activity in the mu/alpha frequency band and extending into the beta frequency band. The time course of activity indicates that visual processing drives subsequent activity in the motor cortex during gesture processing, with a lag of approximately 80 ms. [ABSTRACT FROM AUTHOR]

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An in vivo fluorescent deltorphin (Fluo-DLT) internalization assay was used to assess the distribution and regulation of pharmacologically available delta opioid receptors (deltaORs) in the rat lumbar (L4-5) spinal cord. Under basal conditions, intrathecal injection of Fluo-DLT resulted in the labeling of numerous deltaOR-internalizing neurons throughout dorsal and ventral horns. The distribution and number of Fluo-DLT-labeled perikaryal profiles were consistent with that of deltaOR-expressing neurons, as revealed by in situ hybridization and immunohistochemistry, suggesting that a large proportion of these cells was responsive to intrathecally administered deltaOR agonists. Pretreatment of rats with morphine for 48 hr resulted in a selective increase in Fluo-DLT-labeled perikaryal profiles within the dorsal horn. These changes were not accompanied by corresponding augmentations in either deltaOR mRNA or (125)I-deltorphin-II binding levels, suggesting that they were attributable to higher densities of cell surface deltaOR available for internalization rather than to enhanced production of the receptor. Unilateral dorsal rhizotomy also resulted in increased Fluo-DLT internalization in the ipsilateral dorsal horn when compared with the side contralateral to the deafferentation or to non-deafferented controls, suggesting that deltaOR trafficking in dorsal horn neurons may be regulated by afferent inputs. Furthermore, morphine treatment no longer increased Fluo-DLT internalization on either side of the spinal cord after unilateral dorsal rhizotomy, indicating that microOR-induced changes in the cell surface availability of deltaOR depend on the integrity of primary afferent inputs. Together, these results suggest that regulation of deltaOR responsiveness through microOR activation in this region is linked to somatosensory information processing.

Previous studies from our laboratory have demonstrated that both chronic inflammatory pain, induced by intraplantar injection of complete Freund's adjuvant (CFA), and prolonged (48 h) stimulation of mu-opioid receptors (muOR) by systemic administration of a variety of selective agonists, resulted in enhanced plasma membrane targeting of delta-opioid receptors (deltaOR) in neurons of the dorsal spinal cord. To determine whether deltaOR trafficking induced by chronic inflammation was dependent on the activation of muOR, we investigated by immunogold cytochemistry the effects of intraplantar CFA injection on the plasma membrane density of deltaOR in muOR knockout (KO) mice. In untreated wild-type (WT) mice, only a small proportion of deltaOR was associated with neuronal plasma membranes in the dorsal horn of the spinal cord. The CFA-induced inflammation produced a significantly higher ratio of plasma membrane to intracellular receptors, as well as a 75% increase in the membrane density of immunoreactive deltaOR, in dendrites of the ipsilateral dorsal horn as compared to untreated mice. This increase in the membrane density of deltaOR was likely due to a recruitment of receptors from intracellular stores since no difference in the overall deltaOR immunolabeling density was evident between CFA-treated and untreated mice. Most importantly, the CFA-induced changes in deltaOR plasma membrane insertion seen in WT animals were not present in the spinal cord of muOR KO mice. These results demonstrate that the integrity of muOR is necessary for CFA-induced changes in deltaOR trafficking to occur and suggest that these changes could be elicited by stimulation of muOR by endogenous opioids released in response to chronic inflammatory pain.

An in vivo fluorescent deltorphin (Fluo-DLT) internalization assay was used to assess the distribution and regulation of pharmacologically available delta opioid receptors (deltaORs) in the rat lumbar (L4-5) spinal cord. Under basal conditions, intrathecal injection of Fluo-DLT resulted in the labeling of numerous deltaOR-internalizing neurons throughout dorsal and ventral horns. The distribution and number of Fluo-DLT-labeled perikaryal profiles were consistent with that of deltaOR-expressing neurons, as revealed by in situ hybridization and immunohistochemistry, suggesting that a large proportion of these cells was responsive to intrathecally administered deltaOR agonists. Pretreatment of rats with morphine for 48 hr resulted in a selective increase in Fluo-DLT-labeled perikaryal profiles within the dorsal horn. These changes were not accompanied by corresponding augmentations in either deltaOR mRNA or (125)I-deltorphin-II binding levels, suggesting that they were attributable to higher densities of cell surface deltaOR available for internalization rather than to enhanced production of the receptor. Unilateral dorsal rhizotomy also resulted in increased Fluo-DLT internalization in the ipsilateral dorsal horn when compared with the side contralateral to the deafferentation or to non-deafferented controls, suggesting that deltaOR trafficking in dorsal horn neurons may be regulated by afferent inputs. Furthermore, morphine treatment no longer increased Fluo-DLT internalization on either side of the spinal cord after unilateral dorsal rhizotomy, indicating that microOR-induced changes in the cell surface availability of deltaOR depend on the integrity of primary afferent inputs. Together, these results suggest that regulation of deltaOR responsiveness through microOR activation in this region is linked to somatosensory information processing.

We recently demonstrated that prolonged treatment with morphine increases the antinociceptive potency of the delta-opioid receptor (deltaOR) agonist deltorphin and promotes cell surface targeting of deltaORs in neurons of the dorsal horn of the rat spinal cord (Cahill et al., 2001b). In the present study we examined whether these effects were mediated selectively via muOR. Using the same intermittent treatment regimen as for morphine, we found that methadone and etorphine, but not fentanyl, enhanced [D-Ala2]-deltorphin-mediated antinociception. However, continuous delivery of fentanyl for 48 hr resulted in augmented deltaOR-mediated antinociception when compared with saline-infused animals. Time course studies confirmed that a 48 hr treatment with morphine was necessary for the establishment of enhanced deltaOR-mediated antinociception. The observed increases in deltaOR agonist potency and deltaOR plasma membrane density were reversed fully 48 hr after discontinuation of morphine injections. Wild-type C57BL/6 mice pretreated with morphine for 48 hr similarly displayed enhanced deltaOR-mediated antinociception in a tonic pain paradigm. Accordingly, the percentage of plasma membrane-associated deltaOR in the dorsal horn of the spinal cord, as assessed by immunogold electron microscopy, increased from 6.6% in naive to 12.4% in morphine-treated mice. In contrast, morphine treatment of muOR gene knock-out (KO) mice did not produce any change in deltaOR plasma membrane density. These results demonstrate that selective activation of muOR is critical for morphine-induced targeting of deltaOR to neuronal membranes, but not for basal targeting of this receptor to the cell surface.

We recently demonstrated that prolonged treatment with morphine increases the antinociceptive potency of the delta-opioid receptor (deltaOR) agonist deltorphin and promotes cell surface targeting of deltaORs in neurons of the dorsal horn of the rat spinal cord (Cahill et al., 2001b). In the present study we examined whether these effects were mediated selectively via muOR. Using the same intermittent treatment regimen as for morphine, we found that methadone and etorphine, but not fentanyl, enhanced [D-Ala2]-deltorphin-mediated antinociception. However, continuous delivery of fentanyl for 48 hr resulted in augmented deltaOR-mediated antinociception when compared with saline-infused animals. Time course studies confirmed that a 48 hr treatment with morphine was necessary for the establishment of enhanced deltaOR-mediated antinociception. The observed increases in deltaOR agonist potency and deltaOR plasma membrane density were reversed fully 48 hr after discontinuation of morphine injections. Wild-type C57BL/6 mice pretreated with morphine for 48 hr similarly displayed enhanced deltaOR-mediated antinociception in a tonic pain paradigm. Accordingly, the percentage of plasma membrane-associated deltaOR in the dorsal horn of the spinal cord, as assessed by immunogold electron microscopy, increased from 6.6% in naive to 12.4% in morphine-treated mice. In contrast, morphine treatment of muOR gene knock-out (KO) mice did not produce any change in deltaOR plasma membrane density. These results demonstrate that selective activation of muOR is critical for morphine-induced targeting of deltaOR to neuronal membranes, but not for basal targeting of this receptor to the cell surface.

Molecular sequence data have previously revealed the existence of cryptic species associated with the Pocillopora ‘damicornis-like’ coral. Recently, this species complex has been reclassified into three species including the resurrected P. acuta, which appears to have a wide distribution. Morphological characteristics described for P. acuta are present in corals previously identified as Pocillopora damicornis. To determine if the Pocillopora ‘damicornis-like’ colonies on Singapore’s reefs are P. acuta, P. damicornis, or both, we examined a new collection of Pocillopora using mitochondrial DNA data and morphology. We also compared specimen morphologies from past collections and examined the known regional distributions of both species. Our analyses show that most Pocillopora ‘damicornis-like’ corals in Singapore are P. acuta instead of P. damicornis. Findings here are important for coral diversity records in Singapore and will help clarify distributional limits of morphologically similar Pocillopora species.

Opioid receptors are known to undergo complex regulatory changes in response to ligand exposure. In the present study, we examined the effect of morphine on the in vitro and in vivo density and trafficking of delta opioid receptors (deltaORs). Prolonged exposure (48 hr) of cortical neurons in culture to morphine (10 microm) resulted in a robust increase in the internalization of Fluo-deltorphin, a highly selective fluorescent deltaOR agonist. This effect was mu-mediated because it was entirely blocked by the selective mu opioid receptor antagonist d-Phe-Cys-Tyr-d-Trp-Orn-Thr-Pen-Thr-NH(2) and was reproduced using the selective mu agonist fentanyl citrate. Immunogold electron microscopy revealed a marked increase in the cell surface density of deltaORs in neurons exposed to morphine, indicating that the increase in Fluo-deltorphin internalization was caused by increased receptor availability. Prolonged morphine exposure had no effect on deltaOR protein levels, as assessed by immunocytochemistry and Western blotting, suggesting that the increase in bioavailable deltaORs was caused by recruitment of reserve receptors from intracellular stores and not from receptor neosynthesis. Complementary in vivo studies demonstrated that chronic treatment of adult rats with morphine (5-15 mg/kg, s.c., every 12 hr) similarly augmented targeting of deltaORs to neuronal plasma membranes in the dorsal horn of the spinal cord. Furthermore, this treatment markedly potentiated intrathecal d-[Ala(2)]deltorphin II-induced antinociception. Taken together, these results demonstrate that prolonged stimulation of neurons with morphine markedly increases recruitment of intracellular deltaORs to the cell surface, both in vitro and in vivo. We propose that this type of receptor subtype cross-mobilization may widen the transduction repertoire of G-protein-coupled receptors and offer new therapeutic strategies.

Opioid receptors are known to undergo complex regulatory changes in response to ligand exposure. In the present study, we examined the effect of morphine on the in vitro and in vivo density and trafficking of delta opioid receptors (deltaORs). Prolonged exposure (48 hr) of cortical neurons in culture to morphine (10 microm) resulted in a robust increase in the internalization of Fluo-deltorphin, a highly selective fluorescent deltaOR agonist. This effect was mu-mediated because it was entirely blocked by the selective mu opioid receptor antagonist d-Phe-Cys-Tyr-d-Trp-Orn-Thr-Pen-Thr-NH(2) and was reproduced using the selective mu agonist fentanyl citrate. Immunogold electron microscopy revealed a marked increase in the cell surface density of deltaORs in neurons exposed to morphine, indicating that the increase in Fluo-deltorphin internalization was caused by increased receptor availability. Prolonged morphine exposure had no effect on deltaOR protein levels, as assessed by immunocytochemistry and Western blotting, suggesting that the increase in bioavailable deltaORs was caused by recruitment of reserve receptors from intracellular stores and not from receptor neosynthesis. Complementary in vivo studies demonstrated that chronic treatment of adult rats with morphine (5-15 mg/kg, s.c., every 12 hr) similarly augmented targeting of deltaORs to neuronal plasma membranes in the dorsal horn of the spinal cord. Furthermore, this treatment markedly potentiated intrathecal d-[Ala(2)]deltorphin II-induced antinociception. Taken together, these results demonstrate that prolonged stimulation of neurons with morphine markedly increases recruitment of intracellular deltaORs to the cell surface, both in vitro and in vivo. We propose that this type of receptor subtype cross-mobilization may widen the transduction repertoire of G-protein-coupled receptors and offer new therapeutic strategies.

Background: Microglia activation contributes to chronic pain and to the adverse effects of opiate use such as analgesic tolerance and opioid-induced hyperalgesia. Both mu opioid receptor (MOR) encoded by Oprm1/OPRM1 gene and toll like receptor 4 (TLR4) have been reported to mediate these morphine effects and a current question is whether microglia express the Oprm1 transcript and MOR protein. The aim of this study was to characterize Oprm1-MOR expression in naive murine and human microglia, combining transcriptomics datasets previously published by other groups with our own imaging study using the Cx3cr1-eGFP-MOR-mCherry reporter mouse line.Methods: We analyzed microglial Oprm1/OPRM1 expression obtained from transcriptomics datasets, focusing on ex vivo studies from adult wild-type animals and adult post-mortem human cerebral cortex. Oprm1, as well as co-regulated gene sets were examined. The expression of MOR in microglia was also investigated using our novel fluorescent Cx3cr1-eGFP-MOR-mcherry reporter mouse line. We determined whether CX3cR1-eGFP positive microglial cells expressed MOR-mCherry protein by imaging various brain areas including the Frontal Cortex, Nucleus Accumbens, Ventral Tegmental Area, Central Amygdala, and Periaqueductal Gray matter, as well as spinal cord.Results:Oprm1 expression was found in all 12 microglia datasets from mouse whole brain, in 7 out of 8 from cerebral cortex, 3 out of 4 from hippocampus, 1 out of 1 from striatum, and 4 out of 5 from mouse or rat spinal cord. OPRM1 was expressed in 16 out of 17 microglia transcriptomes from human cerebral cortex. In Cx3cr1-eGFP-MOR-mCherry mice, the percentage of MOR-positive microglial cells ranged between 35.4 and 51.6% in the different brain areas, and between 36.8 and 42.4% in the spinal cord.Conclusion: The comparative analysis of the microglia transcriptomes indicates that Oprm1/OPRM1 transcripts are expressed in microglia. The investigation of Cx3cr1-eGFP-MOR-mCherry mice also shows microglial expression of MOR proteinin the brain and spine. These results corroborate functional studies showing the actions of MOR agonists on microglia and suppression of these effects by MOR-selective antagonists or MOR knockdown.

Background: Intense stress can change pain perception and induce hyperalgesia; a phenomenon called stress-induced hyperalgesia (SIH). However, the neurobiological mechanism of this effect remains unclear. The present study aimed to investigate the effect of the spinal cord µ-opioid receptors (MOR) and α2-adrenergic receptors (α2-AR) on pain sensation in rats with SIH. Methods: Eighteen Sprague-Dawley male rats, weighing 200- 250 g, were randomly divided into two groups (n=9 per group), namely the control and stress group. The stress group was evoked by random 1-hour daily foot-shock stress (0.8 mA for 10 seconds, 1 minute apart) for 3 weeks using a communication box. The tail-flick and formalin tests were performed in both groups on day 22. The real-time RT-PCR technique was used to observe MOR and α2-AR mRNA levels at the L4-L5 lumbar spinal cord. Statistical analysis was performed using the GraphPad Prism 5 software (San Diego, CA, USA). Student's t test was applied for comparisons between the groups. P<0.05 was considered statistically significant. Results: There was a significant (P=0.0014) decrease in tailflick latency in the stress group compared to the control group. Nociceptive behavioral responses to formalin-induced pain in the stress group were significantly increased in the acute (P=0.007) and chronic (P=0.001) phases of the formalin test compared to the control group. A significant reduction was also observed in MOR mRNA level of the stress group compared to the control group (P=0.003). There was no significant difference in α2-AR mRNA level between the stress and control group. Conclusion: The results indicate that chronic stress can affect nociception and lead to hyperalgesia. The data suggest that decreased expression of spinal cord MOR causes hyperalgesia. Please cite this article as: Hormozi A, Zarifkar A, Rostami B, Naghibalhossaini F. An Experimental Study on Spinal Cord µ-Opioid and α2-Adrenergic Receptors mRNA Expression Following Stress-Induced Hyperalgesia in Male Rats. [ABSTRACT FROM AUTHOR]

Introduction: The effects of smoking denicotinized (denic) and average nicotine (avnic) tobacco cigarettes were studied on brain mu opioid receptor binding by positron emission tomography with 11C carfentanil. The results indicated the importance of physiological and psychological effects induced by denic smoking.Methods: Regional mu opioid binding potential (nondisplaceable binding potential, BPND) was measured in 20 adult male overnight abstinent chronic tobacco smokers. The denic sessions were conducted about 8:00 am followed by avnic sessions about 2 hours later. Venous plasma nicotine levels and scores of craving to smoke were assessed before and after each smoking session. Fagerstrom scores of nicotine dependence were determined. Pearson's and Spearman's correlation tests were used to examine associations between BPND and other smoking parameters.Results: Surprisingly, the very low plasma nicotine peak levels after denic smoking (mean ± SD: 3.3 ± 1.8 ng/mL) were significantly correlated with BPND after denic and avnic smoking. Equally surprising no association was found between nicotine levels after avnic smoking and BPND. Delta craving scores and Fagerstrom scores were correlated with both BPND after denic and avnic in several brain regions.Conclusions: Very small amounts of nicotine, psychological and behavioral effects of denic smoking appear to have important actions on the endogenous mu opioid system.Implications: Associations between very low venous plasma nicotine levels after denic smoking and regional brain mu opioid receptor availability are a surprising "placebo" effect. Delta craving and Fagerstrom scores were correlated with BPND in several brain regions including amygdala, hippocampus, insula, nucleus accumbens, putamen, and ventral striatum. This study is limited by modest Power (mean 1 - β = 0.6) for all correlation analyses. [ABSTRACT FROM AUTHOR]

Drugs acting at the opioid receptor family are clinically used to treat chronic and acute pain, though they represent the second line of treatment behind GABA analogs, antidepressants and SSRI's. Within the opioid family mu and kappa opioid receptor are commonly targeted. However, activation of the mu opioid receptor has side effects of constipation, tolerance, dependence, euphoria, and respiratory depression; activation of the kappa opioid receptor leads to dysphoria and sedation. The side effects of mu opioid receptor activation have led to mu receptor drugs being widely abused with great overdose risk. For these reasons, newer safer opioid analgesics are in high demand. For many years a focus within the opioid field was finding drugs that activated the G protein pathway at mu opioid receptor, without activating the β-arrestin pathway, known as biased agonism. Recent advances have shown that this may not be the way forward to develop safer analgesics at mu opioid receptor, though there is still some promise at the kappa opioid receptor. Here we discuss recent novel approaches to develop safer opioid drugs including efficacy vs bias and fine-tuning receptor activation by targeting sub-pockets in the orthosteric site, we explore recent works on the structural basis of bias, and we put forward the suggestion that Gα subtype selectivity may be an exciting new area of interest. [Display omitted] [ABSTRACT FROM AUTHOR]

Morphine releases adenosine from the spinal cord and this contributes to spinal antinociception. The present study examined possible interactions between mu- and subclasses of delta-opioid receptors in the release of adenosine. Nanomolar (10(-8), 10(-9) M) concentrations of morphine release adenosine from spinal cord synaptosomes under conditions of partial depolarization with elevated K+, and this component of release is mediated by activation of mu-opioid receptors. Subnanomolar (10(-10), 10(-11) M) concentrations of the mu-opioid receptor agonists morphine, [N-MePhe3,D-Pro4]morphiceptin, and [D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin (DAMGO) have minimal effects on the release of adenosine from the spinal cord. However, [D-Pen2,D-Pen5]enkephalin (DPDPE), a delta 1-opioid receptor agonist, and [D-Ala2,Cys4]deltorphin, a delta 2-opioid receptor agonist, at doses which exhibit no intrinsic effects (10(-8) and 10(-7) M), shifted the dose-response curve for mu-opioid receptor-evoked adenosine release to the left in a dose-dependent manner. DPDPE was more potent than [D-Ala2,Cys4]deltorphin when combined with the highly selective mu-opioid receptor agonist [N-MePhe3,D-Pro4]morphiceptin, but these agents showed similar activity with the less selective agonists DAMGO and morphine. Simultaneous activation of mu- and delta-opioid receptors generates a synergistic release of adenosine from spinal cord synaptosomes. Although agonists for both delta 1- and delta 2-opioid receptor subtypes produce this response, the delta 1-opioid receptor agonist is more potent at eliciting this effect when the most selective mu-opioid receptor ligand is used.

Morphine releases adenosine from the spinal cord and this contributes to spinal antinociception. The present study examined possible interactions between mu- and subclasses of delta-opioid receptors in the release of adenosine. Nanomolar (10(-8), 10(-9) M) concentrations of morphine release adenosine from spinal cord synaptosomes under conditions of partial depolarization with elevated K+, and this component of release is mediated by activation of mu-opioid receptors. Subnanomolar (10(-10), 10(-11) M) concentrations of the mu-opioid receptor agonists morphine, [N-MePhe3,D-Pro4]morphiceptin, and [D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin (DAMGO) have minimal effects on the release of adenosine from the spinal cord. However, [D-Pen2,D-Pen5]enkephalin (DPDPE), a delta 1-opioid receptor agonist, and [D-Ala2,Cys4]deltorphin, a delta 2-opioid receptor agonist, at doses which exhibit no intrinsic effects (10(-8) and 10(-7) M), shifted the dose-response curve for mu-opioid receptor-evoked adenosine release to the left in a dose-dependent manner. DPDPE was more potent than [D-Ala2,Cys4]deltorphin when combined with the highly selective mu-opioid receptor agonist [N-MePhe3,D-Pro4]morphiceptin, but these agents showed similar activity with the less selective agonists DAMGO and morphine. Simultaneous activation of mu- and delta-opioid receptors generates a synergistic release of adenosine from spinal cord synaptosomes. Although agonists for both delta 1- and delta 2-opioid receptor subtypes produce this response, the delta 1-opioid receptor agonist is more potent at eliciting this effect when the most selective mu-opioid receptor ligand is used.

Release of adenosine from the spinal cord contributes to spinal antinociception by morphine. Morphine induces a Ca(++)-dependent release of adenosine from dorsal spinal cord synaptosomes, which is augmented under partially depolarizing conditions. The present study examined the opioid receptor subtypes involved in this release, and determined whether adenosine is an important mediator of antinociception induced by the spinal administration of selective opioid agonists in rats. Nanomolar and micromolar concentrations of the selective mu opioid agonists DAMGO ([D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin) and PLO17 ([N-MePhe3,D-Pro4]morphiceptin) induced release of adenosine in a biphasic manner in the presence of a partial depolarization (addition of 6 mM K+ to the Krebs' medium). The delta opioid agonists DPDPE ([D-Pen2,D-Pen5]enkephalin) and DELT ([D-Ala2,Cys4]deltorphin) and the kappa opioid agonist U50488H (trans-(+/-)-3,4-dichloro-N-methyl-N-(2-(1-pyrroli-zemeacetamid e) had little effect on the release of adenosine except at high micromolar concentrations. Release of adenosine by mu (nanomolar) and delta (micromolar) ligands is Ca(++)-dependent, whereas the kappa (micromolar) receptor ligand releases adenosine via a Ca(++)-independent mechanism. Behavioral antinociception using the hot-plate threshold test revealed that intrathecal administration of the mu and delta opioid receptor agonists produced dose-dependent antinociception with an order of potency of DAMGO, PLO17 > morphine, DELT > DPDPE. An ED75 dose of morphine, DAMGO or PLO17 was attenuated dose-dependently by intrathecal pretreatment with the adenosine receptor antagonist caffeine. Caffeine did not block the antinociceptive response to delta agonists, but in fact augmented antinociception when combined with DPDPE and DELT. This augmentation was dose-dependent. This study demonstrates that activation of the mu receptor subtype is responsible for the opioid-induced release of adenosine from the spinal cord, that such release contributes to the spinal antinociception by mu agonists and that only release evoked by low doses of opioids is behaviorally relevant.

Competition changes the environment for athletes. The difficulty of training for such stressful events can lead to the well-known effect of “choking” under pressure, which prevents athletes from performing at their best level. To study the effect of competition on the human brain, we recorded pilot electroencephalography (EEG) data while novice shooters were immersed in a realistic virtual environment representing a shooting range. We found a differential between-subject effect of competition on mu (8–12 Hz) oscillatory activity during aiming; compared to training, the more the subject was able to desynchronize his mu rhythm during competition, the better was his shooting performance. Because this differential effect could not be explained by differences in simple measures of the kinematics and muscular activity, nor by the effect of competition or shooting performance per se, we interpret our results as evidence that mu desynchronization has a positive effect on performance during competition.

Abstract The mere perception of high-calorie food items can trigger strong action tendencies towards these foods. Go/no-go training has successfully been applied to reduce such action tendencies. This study investigated the electrophysiological mechanisms that may underlie the beneficial effects of go/no-go training on food consumption. EEG was measured while 19 participants passively observed pictures of food and non-food items, both before and after the go/no-go training. During training, 50% of the food and non-food items were consistently paired with a go/no-go response. After training, food items that had been associated with a response induced larger mu desynchronization at electrodes over sensorimotor regions, whereas food items that had been associated with withholding from responding induced larger increases in theta power at frontal midline electrodes. These findings suggest that the exerted cognitive control during go/no-go training with attractive food stimuli may become associated with these stimuli and signal the required level of control during subsequent encounters. Highlights • Go/no-go training affects food item processing during subsequent passive viewing. • Frontal theta power is larger for food items paired with no-go than go responses. • Mu desynchronization is higher for food items paired with go than no-go responses. • The need for cognitive control seems to be tagged to specific stimuli. [ABSTRACT FROM AUTHOR]

Competition changes the environment for athletes. The difficulty of training for such stressful events can lead to the well-known effect of "choking" under pressure, which prevents athletes from performing at their best level. To study the effect of competition on the human brain, we recorded pilot electroencephalography (EEG) data while novice shooters were immersed in a realistic virtual environment representing a shooting range. We found a differential between-subject effect of competition on mu (8-12 Hz) oscillatory activity during aiming; compared to training, the more the subject was able to desynchronize his mu rhythm during competition, the better was his shooting performance. Because this differential effect could not be explained by differences in simple measures of the kinematics and muscular activity, nor by the effect of competition or shooting performance per se , we interpret our results as evidence that mu desynchronization has a positive effect on performance during competition. [ABSTRACT FROM AUTHOR]

The performance of asphalt concrete pavement has affected by many factors, the temperature is the most important environmental one which has a large effect on the structural behavior of flexible pavement materials. The main cause of premature failure of pavement is the rutting, Due to the viscoelastic nature of the asphalt cement, rutting is more pronounced in hot climate areas because the viscosity of the asphalt binder which is inversely related to rutting is significantly reduced with the increase in temperature resulting in a more rut susceptible paving mixtures. The objective of this study is to determine the effect of temperatures variations on the permanent deformation parameters (permanent strain (p), intercept (a), slope (b), Alpha and Mu) as well as resilient strain (r) and resilient modulus (Mr). To achieve this objective, one aggregate gradation with 12.5mm nominal maximum size, two grades of asphalt cements (40-50 and 60-70) brought form Al- Daurah refinery, limestone dust filler has been used to prepare the asphalt concrete mixtures. 30 Marshall specimens were prepared to determine the optimum asphalt cement content. Thereafter, 30 cylindrical asphalt concrete specimens (102mm in diameter and 203 mm in height) are prepared in optimum asphalt cement and optimum ±0.5 percent. The prepared specimens were used in uniaxial repeated load test to evaluate the permanent deformation parameters of asphalt concrete mixes under the following testing temperature (5, 15, 25, 40 and 60c). The test result analyses appeared that Mr is decrease 51 percent when temperature increased from 5 c to 25 c and then decrease 22 percent with further increase in temperature from 25 c to 60 c. Also, the Alpha value decreases by a factor of 1.25 and 1.13 when temperature increases from 5 c to 25 c and 25 c to 60 c, espectively. Finally, statistical models were developed to predict the Alpha and Mu parameters of permanent deformation.

The transposition of bacteriophage Mu serves as a model system for understanding DDE transposases and integrases. All available structures of these enzymes at the end of the transposition reaction, including Mu, exhibit significant bends in the transposition target site DNA. Here we use Mu to investigate the ramifications of target DNA bending on the transposition reaction. Enhancing the flexibility of the target DNA or prebending it increases its affinity for transpososomes by over an order of magnitude and increases the overall reaction rate. This and FRET confirm that flexibility is interrogated early during the interaction between the transposase and a potential target site, which may be how other DNA binding proteins can steer selection of advantageous target sites. We also find that the conformation of the target DNA after strand transfer is involved in preventing accidental catalysis of the reverse reaction, as conditions that destabilize this conformation also trigger reversal.

Background Autism-like symptoms in the non-clinical general population are referred to as the broad autism phenotype (BAP). To date there have been no studies investigating how BAP might correlate with measurements from the resting-state electroencephalogram (EEG). Method EEG resting-state data were collected in 20 young adults during both eyes-closed (EC) and eyes-open (EO) resting states. Permutation modelling was used to assess correlations of the Broad Autism Phenotype Questionnaire (BAPQ) with source localised resting alpha activity. Results Total scores on the BAPQ were strongly correlated with differences between oscillatory brain activity during EC and EO rest in a pattern that was classified as the mu rhythm. More mu activity during EC rest compared to EO rest was found to be associated with higher BAPQ scores (i.e. more prominent BAP symptoms). Conclusions Mu is a known correlate of activity in the mirror neuron system (MNS), which has been implicated in the social deficits associated with autism. It is therefore suggested that this BAPQ-correlated mu activity could reflect the readying of the MNS for social input by visual stimulation. These findings offer novel insights into how the BAP is reflected in resting-state recordings of brain activity. [ABSTRACT FROM AUTHOR]

Background: Nicotine addiction continues to be a health challenge across the world. Despite several approved medications, smokers continue to relapse. Several human and animal studies have evaluated the role of the endogenous opioid system as a potential target for smoking cessation medications. Methods: In this review, studies that have elucidated the role of the mu (MORs), delta (DORs), and kappa (KORs) opioid receptors in nicotine reward, nicotine withdrawal, and reinstatement of nicotine seeking will be discussed. Additionally, the review will discuss discrepancies in the literature and therapeutic potential of the endogenous opioid system, and suggest studies to address gaps in knowledge with respect to the role of the opioid receptors in nicotine dependence. Results: Data available till date suggest that blockade of the MORs and DORs decreased the rewarding effects of nicotine, while activation of the MORs and DORs decreased nicotine withdrawal-induced aversive effects. In contrast, activation of the KORs decreased the rewarding effects of nicotine, while blockade of the KORs decreased nicotine withdrawal-induced aversive effects. Interestingly, blockade of the MORs and KORs attenuated reinstatement of nicotine seeking. In humans, MOR antagonists have shown benefits in select subpopulations of smokers and further investigation is required to realize their full therapeutic potential. Conclusion: Future work must assess the influence of polymorphisms in opioid receptor-linked genes in nicotine dependence, which will help in both identifying individuals vulnerable to nicotine addiction and the development of opioid-based smoking cessation medications. Overall, the endogenous opioid system continues to be a promising target for future smoking cessation medications. [ABSTRACT FROM AUTHOR]