Search

Excessive fructose intake is a risk factor for the development of obesity and its complications. Targeting ketohexokinase (KHK), the first enzyme of fructose metabolism, has been investigated for the management of MASLD. We compared the effects of systemic, small molecule inhibitor of KHK enzymatic activity to hepatocyte-specific, GalNAc-siRNA mediated knockdown of KHK in mice on a HFD. We measured KHK enzymatic activity, extensively quantified glycogen accumulation, performed RNAseq analysis, and enumerated hepatic metabolites using mass spectrometry. Both KHK siRNA and KHK inhibitor led to an improvement in liver steatosis, however, via substantially different mechanisms. KHK knockdown decreased the de novo lipogenesis pathway, whereas the inhibitor increased the fatty acid oxidation pathway. Moreover, KHK knockdown completely prevented hepatic fructolysis and improved glucose tolerance. Conversely, the KHK inhibitor only partially reduced fructolysis, but it also targeted triokinase, mediating the third step of fructolysis. This leads to the accumulation of fructose-1 phosphate, resulting in glycogen accumulation, hepatomegaly, and impaired glucose tolerance. Overexpression of wild-type, but not kinase-dead KHK in cultured hepatocytes increased hepatocyte injury and glycogen accumulation when treated with fructose. The differences between KHK inhibition and knockdown are, in part, explained by the kinase-dependent and independent effects of KHK on hepatic metabolism.

Substance use disorders, including alcohol use disorder, are a public health concern that affect more than 150 million people globally. The opioid antagonist naltrexone is being increasingly prescribed to treat opioid use disorder, alcohol use disorder, and chronic pain. Perioperative management of patients on naltrexone is inconsistent and remains a controversial topic, with mismanagement posing a significant risk to the long-term health of these patients. This scoping review was conducted to identify human studies in which the perioperative management of naltrexone was described. This review includes a systematic literature search involving Medline, Medline In-Process, Embase, PsycINFO, and Web of Science. Seventeen articles that describe perioperative naltrexone management strategies were included, including thirteen guidelines, one case report, and three randomized trials. Despite its use in patients with alcohol use disorder and chronic pain, no clinical studies, case reports, or guidelines addressed naltrexone use in these clinical populations. All of the guideline documents recommended the preoperative cessation of naltrexone, irrespective of dose, indication, or route of administration. None of these guideline documents were designed on the basis of a systematic literature search or a Delphi protocol. As described by the primary studies, perioperative pain relief varied depending on naltrexone dose and route of administration, time since last naltrexone administration, and underlying substance use disorder. None of the studies commented on the maintenance of recovery for the patient's substance use disorder in the context of perioperative naltrexone management. The current understanding of the risks and benefits of continuing or stopping naltrexone perioperatively is limited by a lack of high-quality evidence. In patients with risk factors for return to use of opioids or alcohol, the discontinuation of naltrexone should have a strong rationale. Future studies and guidelines should seek to address both acute pain management and maintaining recovery when discussing perioperative naltrexone management strategies., (Copyright © 2024 American Society of Anesthesiologists. All Rights Reserved.)

Dopaminergic signaling in the nucleus accumbens shell (NAc) regulates neuronal activity relevant to reward-related learning, including cocaine-associated behaviors. Although astrocytes respond to dopamine and cocaine with structural changes, the impact of dopamine and cocaine on astrocyte functional plasticity has not been widely studied. Specifically, behavioral implications of voltage-gated channel activity in the canonically non-excitable astrocytes are not known. We characterized potassium channel function in NAc astrocytes following exposure to exogenous dopamine or cocaine self-administration training under short (2 h/day) and extended (6 h/day) access schedules. Electrophysiological, Ca 2+ imaging, mRNA, and mass spectrometry tools were used for molecular characterization. Behavioral effects were examined after NAc-targeted microinjections of channel antagonists and astroglial toxins. Exogenous dopamine increased activity of currents mediated by voltage-gated (K v 7) channels in NAc astrocytes. This was associated with a ~5-fold increase in expression of Kcnq2 transcript level in homogenized NAc micropunches. Matrix-assisted laser desorption/ionization mass spectrometry revealed increased NAc dopamine levels in extended access, relative to short access, rats. K v 7 inhibition selectively increased frequency and amplitude of astrocyte intracellular Ca 2+ transients in NAc of extended access rats. Inhibition of K v 7 channels in the NAc attenuated cocaine-seeking in extended access rats only, an effect that was occluded by microinjection of the astrocyte metabolic poison, fluorocitrate. These results suggest that voltage-gated K + channel signaling in NAc astrocytes is behaviorally relevant, support K v 7-mediated regulation of astrocyte Ca 2+ signals, and propose novel mechanisms of neuroglial interactions relevant to drug use., (© 2023. The Author(s), under exclusive licence to American College of Neuropsychopharmacology.)

Background and Aims: Genome and epigenome wide association studies identified variants in carnitine palmitoyltransferase 1a (CPT1a) that associate with lipid traits. The goal of this study was to determine the role of liver-specific CPT1a on hepatic lipid metabolism., Approach and Results: Male and female liver-specific knockout (LKO) and littermate controls were placed on a low-fat or high-fat diet (60% kcal fat) for 15 weeks. Mice were necropsied after a 16 h fast, and tissues were collected for lipidomics, matrix-assisted laser desorption ionization mass spectrometry imaging, kinome analysis, RNA-sequencing, and protein expression by immunoblotting. Female LKO mice had increased serum alanine aminotransferase levels which were associated with greater deposition of hepatic lipids, while male mice were not affected by CPT1a deletion relative to male control mice. Mice with CPT1a deletion had reductions in DHA-containing phospholipids at the expense of monounsaturated fatty acids (MUFA)-containing phospholipids in whole liver and at the level of the lipid droplet (LD). Male and female LKO mice increased RNA levels of genes involved in LD lipolysis (Plin2, Cidec, G0S2) and in polyunsaturated fatty acid metabolism (Elovl5, Fads1, Elovl2), while only female LKO mice increased genes involved in inflammation (Ly6d, Mmp12, Cxcl2). Kinase profiling showed decreased protein kinase A activity, which coincided with increased PLIN2, PLIN5, and G0S2 protein levels and decreased triglyceride hydrolysis in LKO mice., Conclusions: Liver-specific deletion of CPT1a promotes sexually dimorphic steatotic liver disease (SLD) in mice, and here we have identified new mechanisms by which females are protected from HFD-induced liver injury., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Patients with Lafora disease have a mutation in EPM2A or EPM2B, resulting in dysregulation of glycogen metabolism throughout the body and aberrant glycogen molecules that aggregate into Lafora bodies. Lafora bodies are particularly damaging in the brain, where the aggregation drives seizures with increasing severity and frequency, coupled with neurodegeneration. Previous work employed mouse genetic models to reduce glycogen synthesis by approximately 50%, and this strategy significantly reduced Lafora body formation and disease phenotypes. Therefore, an antisense oligonucleotide (ASO) was developed to reduce glycogen synthesis in the brain by targeting glycogen synthase 1 (Gys1). To test the distribution and efficacy of this drug, the Gys1-ASO was administered to Epm2b-/- mice via intracerebroventricular administration at 4, 7, and 10 months. The mice were then sacrificed at 13 months and their brains analyzed for Gys1 expression, glycogen aggregation, and neuronal excitability. The mice treated with Gys1-ASO exhibited decreased Gys1 protein levels, decreased glycogen aggregation, and reduced epileptiform discharges compared to untreated Epm2b-/- mice. This work provides proof of concept that a Gys1-ASO halts disease progression of EPM2B mutations of Lafora disease., (© 2023. The Author(s).)

Oligodendrocytes (OLs) generate lipid-rich myelin membranes that wrap axons to enable efficient transmission of electrical impulses. Using a RIT1 knockout mouse model and in situ high-resolution matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) coupled with MS-based lipidomic analysis to determine the contribution of RIT1 to lipid homeostasis. Here, we report that RIT1 loss is associated with altered lipid levels in the central nervous system (CNS), including myelin-associated lipids within the corpus callosum (CC). Perturbed lipid metabolism was correlated with reduced numbers of OLs, but increased numbers of GFAP + glia, in the CC, but not in grey matter. This was accompanied by reduced myelin protein expression and axonal conduction deficits. Behavioral analyses revealed significant changes in voluntary locomotor activity and anxiety-like behavior in RIT1 KO mice. Together, these data reveal an unexpected role for RIT1 in the regulation of cerebral lipid metabolism, which coincide with altered white matter tract oligodendrocyte levels, reduced axonal conduction velocity, and behavioral abnormalities in the CNS., Competing Interests: Ramon C. Sun has received research support and has received a consultancy fee from Maze Therapeutics. Matthew S. Gentry has received research support and research compounds from Maze Therapeutics, Valerion Therapeutics, and Ionis Pharmaceuticals. Matthew S. Gentry also received a consultancy fee from Maze Therapeutics, PTC Therapeutics, and the Glut1-Deficiency Syndrome Foundation. Fang Wang, Lei Wu, Mritunjay Pandey, Harrison A. Clarke, Hilaree N. Frazier, Carole L. Moncman, Weikang Cai, Lyndsay E.A. Young, Olivier Thibault, and Douglas A. Andres report no disclosures. The content is the responsibility of the authors and does not necessarily represent the official views of the NIH. The paper is subject to the NIH Public Access Policy. This study was carried out in accordance with the Uniform Requirements for Manuscripts Submitted to Biomedical Journals., (© 2023 The Authors. Published by Elsevier Ltd.)

Matrix assisted laser desorption/ionization imaging has greatly improved our understanding of spatial biology, however a robust bioinformatic pipeline for data analysis is lacking. Here, we demonstrate the application of high-dimensionality reduction/spatial clustering and histopathological annotation of matrix assisted laser desorption/ionization imaging datasets to assess tissue metabolic heterogeneity in human lung diseases. Using metabolic features identified from this pipeline, we hypothesize that metabolic channeling between glycogen and N-linked glycans is a critical metabolic process favoring pulmonary fibrosis progression. To test our hypothesis, we induced pulmonary fibrosis in two different mouse models with lysosomal glycogen utilization deficiency. Both mouse models displayed blunted N-linked glycan levels and nearly 90% reduction in endpoint fibrosis when compared to WT animals. Collectively, we provide conclusive evidence that lysosomal utilization of glycogen is required for pulmonary fibrosis progression. In summary, our study provides a roadmap to leverage spatial metabolomics to understand foundational biology in pulmonary diseases., (© 2023. The Author(s).)

High-cervical spinal cord injury often disrupts respiratory motor pathways and disables breathing in the affected population. Moreover, cervically injured individuals are at risk for developing acute lung injury, which predicts substantial mortality rates. While the correlation between acute lung injury and spinal cord injury has been found in the clinical setting, the field lacks an animal model to interrogate the fundamental biology of this relationship. To begin to address this gap in knowledge, we performed an experimental cervical spinal cord injury (N = 18 ) alongside sham injury ( N = 3) and naïve animals ( N = 15) to assess lung injury in adult rats. We demonstrate that animals display some early signs of lung injury two weeks post-spinal cord injury. While no obvious histological signs of injury were observed, the spinal cord injured cohort displayed significant signs of metabolic dysregulation in multiple pathways that include amino acid metabolism, lipid metabolism, and N-linked glycosylation. Collectively, we establish for the first time a model of lung injury after spinal cord injury at an acute time point that can be used to monitor the progression of lung damage, as well as identify potential targets to ameliorate acute lung injury., Competing Interests: The authors report no competing interests., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Guarantors of Brain.)

The E4 allele of Apolipoprotein E (APOE) is associated with both metabolic dysfunction and a heightened pro-inflammatory response: two findings that may be intrinsically linked through the concept of immunometabolism. Here, we combined bulk, single-cell, and spatial transcriptomics with cell-specific and spatially resolved metabolic analyses in mice expressing human APOE to systematically address the role of APOE across age, neuroinflammation, and AD pathology. RNA sequencing (RNA-seq) highlighted immunometabolic changes across the APOE4 glial transcriptome, specifically in subsets of metabolically distinct microglia enriched in the E4 brain during aging or following an inflammatory challenge. E4 microglia display increased Hif1α expression and a disrupted tricarboxylic acid (TCA) cycle and are inherently pro-glycolytic, while spatial transcriptomics and mass spectrometry imaging highlight an E4-specific response to amyloid that is characterized by widespread alterations in lipid metabolism. Taken together, our findings emphasize a central role for APOE in regulating microglial immunometabolism and provide valuable, interactive resources for discovery and validation research., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Glycogen dysregulation is a hallmark of aging, and aberrant glycogen drives metabolic reprogramming and pathogenesis in multiple diseases. However, glycogen heterogeneity in healthy and diseased tissues remains largely unknown. Herein, we describe a method to define spatial glycogen architecture in mouse and human tissues using matrix-assisted laser desorption/ionization mass spectrometry imaging. This assay provides robust and sensitive spatial glycogen quantification and architecture characterization in the brain, liver, kidney, testis, lung, bladder, and even the bone. Armed with this tool, we interrogated glycogen spatial distribution and architecture in different types of human cancers. We demonstrate that glycogen stores and architecture are heterogeneous among diseases. Additionally, we observe unique hyperphosphorylated glycogen accumulation in Ewing sarcoma, a pediatric bone cancer. Using preclinical models, we correct glycogen hyperphosphorylation in Ewing sarcoma through genetic and pharmacological interventions that ablate in vivo tumor growth, demonstrating the clinical therapeutic potential of targeting glycogen in Ewing sarcoma., (© 2022 The Authors. Published under the terms of the CC BY 4.0 license.)

N-linked protein glycosylation in the brain is an understudied facet of glucose utilization that impacts a myriad of cellular processes including resting membrane potential, axon firing, and synaptic vesicle trafficking. Currently, a spatial map of N-linked glycans within the normal and Alzheimer's disease (AD) human brain does not exist. A comprehensive analysis of the spatial N-linked glycome would improve our understanding of brain energy metabolism, linking metabolism to signaling events perturbed during AD progression, and could illuminate new therapeutic strategies. Herein we report an optimized in situ workflow for enzyme-assisted, matrix-assisted laser desorption and ionization (MALDI) mass spectrometry imaging (MSI) of brain N-linked glycans. Using this workflow, we spatially interrogated N-linked glycan heterogeneity in both mouse and human AD brains and their respective age-matched controls. We identified robust regional-specific N-linked glycan changes associated with AD in mice and humans. These data suggest that N-linked glycan dysregulation could be an underpinning of AD pathologies., (© 2021 the Alzheimer's Association.)

Objective: With the increasing prevalence of cannabis use, there is a growing concern about its association with depression and suicidality. The aim of this study was to examine the relationship between recent cannabis use and suicidal ideation using a nationally representative data set., Methods: A cross-sectional analysis of adults was undertaken using National Health and Nutrition Examination Survey data from 2005 to 2018. Participants were dichotomized by whether or not they had used cannabis in the past 30 days. The primary outcome was suicidal ideation, and secondary outcomes were depression and having recently seen a mental health professional. Multiple logistic regression was used to adjust for potential confounders, and survey sample weights were considered in the model., Results: Compared to those with no recent use ( n = 18,599), recent users ( n = 3,127) were more likely to have experienced suicidal ideation in the past 2 weeks (adjusted odds ratio [aOR] 1.54, 95% CI, 1.19 to 2.00, P = 0.001), be depressed (aOR 1.53, 95% CI, 1.29 to 1.82, P < 0.001), and to have seen a mental health professional in the past 12 months (aOR 1.28, 95% CI, 1.04 to 1.59, P = 0.023)., Conclusions: Cannabis use in the past 30 days was associated with suicidal thinking and depression in adults. This relationship is likely multifactorial but highlights the need for specific guidelines and policies for the prescription of medical cannabis for psychiatric therapy. Future research should continue to characterize the health effects of cannabis use in the general population.

Background: Persistent post-surgical pain is an important and under-recognised problem that is difficult to treat. Postoperative complications have been identified as possible risk factors for persistent post-surgical pain. We conducted a secondary analysis of the Measurement of Exercise Tolerance before Surgery (METS) cohort study to characterise the association of major postoperative complications with post-surgical pain at 30 days and 1 yr after major surgery., Methods: The analysis included 1313 participants (≥40 yr old) who had inpatient noncardiac surgery and survived for 1 yr. The co-primary outcomes were 30-day post-surgical pain and 1-yr post-surgical pain. Post-surgical pain was defined as pain or discomfort that was of moderate or severe intensity (EuroQoL-5D [EQ-5D] instrument) and unimproved compared with preoperative pain or discomfort. The principal exposure was major in-hospital complications (moderate or severe by modified Clavien-Dindo criteria). Multivariable logistic regression modelling was used to characterise the adjusted association of major complications with outcomes., Results: Of the cohort, 12% (n=163) experienced major complications, 51% (n=674) reported 30-day post-surgical pain, and 42% (n=545) reported 1-yr post-surgical pain. Major complications were associated with 30-day post-surgical pain (adjusted odds ratio [aOR]=1.54; 95% confidence interval [CI], 1.05-2.23) and possibly 1-yr post-surgical pain (aOR=1.42; 95% CI, 0.98-2.06). When analyses were repeated after multiple imputation of missing covariate and outcome data, complications were associated with both 30-day and 1-yr post-surgical pain., Conclusions: Patients who developed major complications were more likely to report pain at 30 days and possibly 1 yr after surgery. Research is necessary to validate these findings and delineate underlying mechanisms., (Copyright © 2021 British Journal of Anaesthesia. Published by Elsevier Ltd. All rights reserved.)

The tumor microenvironment contains a heterogeneous population of stromal and cancer cells that engage in metabolic crosstalk to ultimately promote tumor growth and contribute to progression. Due to heterogeneity within solid tumors, pooled mass spectrometry workflows are less sensitive at delineating unique metabolic perturbations between stromal and immune cell populations. Two critical, but understudied, facets of glucose metabolism are anabolic pathways for glycogen and N-linked glycan biosynthesis. Together, these complex carbohydrates modulate bioenergetics and protein-structure function, and create functional microanatomy in distinct cell populations within the tumor heterogeneity. Herein, we combine high-dimensionality reduction and clustering (HDRC) analysis with matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) and demonstrate its ability for the comprehensive assessment of tissue histopathology and metabolic heterogeneity in human FFPE sections. In human lung adenocarcinoma (LUAD) tumor tissues, HDRC accurately clusters distinct regions and cell populations within the tumor microenvironment, including tumor cells, tumor-infiltrating lymphocytes, cancer-associated fibroblasts, and necrotic regions. In-depth pathway enrichment analyses revealed unique metabolic pathways are associated with each distinct pathological region. Further, we highlight the potential of HDRC analysis to study complex carbohydrate metabolism in a case study of lung cancer disparity. Collectively, our results demonstrate the promising potentials of HDRC of pixel-based carbohydrate analysis to study cell-type and regional-specific stromal signaling within the tumor microenvironment., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Prostate cancer is the most common cancer in men worldwide. Despite its prevalence, there is a critical knowledge gap in understanding factors driving disparities in survival among different cohorts of patients with prostate cancer. Identifying molecular features separating disparate populations is an important first step in prostate cancer research that could lead to fundamental hypotheses in prostate biology, predictive biomarker discovery, and personalized therapy. N-linked glycosylation is a cotranslational event during protein folding that modulates a myriad of cellular processes. Recently, aberrant N-linked glycosylation has been reported in prostate cancers. However, the full clinical implications of dysregulated glycosylation in prostate cancer has yet to be explored. Herein, we performed direct on-tissue analysis of N-linked glycans using matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) from tissue microarrays of over 100 patient tumors with over 10 years of follow-up metadata. We successfully identified a panel of N-glycans that are unique between benign and prostate tumor tissue. Specifically, high-mannose as well as tri-and tetra-antennary N-glycans were more abundant in tumor tissue and increase proportionally with tumor grade. Further, we expanded our analyses to examine the N-glycan profiles of Black and Appalachian patients and have identified unique glycan signatures that correlate with recurrence in each population. Our study highlights the potential applications of MALDI-MSI for digital pathology and biomarker discovery for prostate cancer. IMPLICATIONS: MALDI-MSI identifies N-glycan perturbations in prostate tumors compared with benign tissue. This method can be utilized to predict prostate cancer recurrence and study prostate cancer disparities., (©2021 American Association for Cancer Research.)

Glycosylation defects are a hallmark of many nervous system diseases. However, the molecular and metabolic basis for this pathology is not fully understood. In this study, we found that N-linked protein glycosylation in the brain is metabolically channeled to glucosamine metabolism through glycogenolysis. We discovered that glucosamine is an abundant constituent of brain glycogen, which functions as a glucosamine reservoir for multiple glycoconjugates. We demonstrated the enzymatic incorporation of glucosamine into glycogen by glycogen synthase, and the release by glycogen phosphorylase by biochemical and structural methodologies, in primary astrocytes, and in vivo by isotopic tracing and mass spectrometry. Using two mouse models of glycogen storage diseases, we showed that disruption of brain glycogen metabolism causes global decreases in free pools of UDP-N-acetylglucosamine and N-linked protein glycosylation. These findings revealed fundamental biological roles of brain glycogen in protein glycosylation with direct relevance to multiple human diseases of the central nervous system., Competing Interests: Declaration of interests M.S.G. is a consultant for Maze Therapeutics, Enable Therapeutics, Glut1-Deficiency Syndrome Foundation, and Chelsea's Hope. M.S.G., R.C.S., C.W.V.K., and R.C.B. are founders of Atterogen, LLC., (Copyright © 2021 Elsevier Inc. All rights reserved.)

N-glycans and lipids are structural metabolites that play important roles in cellular processes. Both show unique regional distribution in tissues; therefore, spatial analyses of these metabolites are crucial to our understanding of cellular physiology. Matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) is an innovative technique that enables in situ detection of analytes with spatial distribution. This workflow details a MALDI-MSI protocol for the spatial profiling of N-glycans and lipids from tissues following application of enzyme and MALDI matrix. For complete details on the use and execution of this protocol, please refer to Drake et al. (2018) and Andres et al. (2020)., Competing Interests: The authors declare no competing interests., (© 2021 The Author(s).)

Cannabis is widely used as a therapeutic drug, especially by patients suffering from psychiatric and neurodegenerative diseases. However, the complex interplay between phytocannabinoids and their targets in the human receptome remains largely a mystery, and there have been few investigations into the relationship between the chemical composition of medical cannabis and the corresponding biological activity. In this study, we investigated 59 cannabis samples used by patients for medical reasons. The samples were subjected to extraction (microwave and supercritical carbon dioxide) and chemical analyses, and the resulting extracts were assayed in vitro using the CB 1 and CB 2 receptors. Using a partial least squares regression analysis, the chemical compositions of the extracts were then correlated to their corresponding cannabinoid receptor activities, thus generating predictive models that describe the receptor potency as a function of major phytocannabinoid content. Using the current dataset, meaningful models for CB 1 and CB 2 receptor agonism were obtained, and these reveal the insignificant relationships between the major phytocannabinoid content and receptor affinity for CB 1 but good correlations between the two at CB 2 receptors. These results also explain the anomalies between the receptor activities of pure phytocannabinoids and cannabis extracts. Furthermore, the models for CB 1 and CB 2 agonism in cannabis extracts predict the cannabinoid receptor activities of individual phytocannabinoids with reasonable accuracy. Here for the first time, we disclose a method to predict the relationship between the chemical composition, including phytocannabinoids, of cannabis extracts and cannabinoid receptor responses.

Importance: The prescription of opioids at discharge after abdominopelvic surgery is variable and often excessive. A lack of guidance for abdominopelvic surgeons may explain the suboptimal nature of current prescribing practices., Objective: To systematically review existing recommendations on the prescription of opioids at discharge, the appropriate disposal of opioids, and the prevention of chronic postsurgical opioid use after abdominopelvic surgery., Evidence Review: This systematic review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. From January 2010 to December 2018, a search of MEDLINE, PsycINFO, HealthSTAR, Embase, and the difficult to locate and unpublished (ie, gray) literature was performed using a peer-reviewed strategy with variations of the terms opioid, surgery, and guideline to identify English-language documents that contained recommendations published by professional societies or health care institutions. The quality of clinical practice guidelines was assessed using the Appraisal of Guidelines Research and Evaluation II (AGREE II) tool. A descriptive synthesis of results was performed., Findings: Of 5530 citations screened, 41 full-text documents were included in the systematic review. Fifteen clinical practice guidelines were identified. AGREE II domain scores varied substantially. Identified among the 41 included documents were 98 recommended interventions for the prescription of opioids at discharge, 8 interventions for the disposal of opioids, and 8 interventions for the prevention of chronic postsurgical opioid use. Only 13 of 114 interventions (11.4%) were supported by an assessment of strength or level of evidence, and the amount of opioid recommended after specific abdominopelvic surgical procedures varied widely between guidance documents, even for the same procedure., Conclusions and Relevance: Current guidance for the prescription of opioids at discharge after abdominopelvic surgery is heterogeneous and rarely supported by evidence. More research is needed on this topic to guide the development of future recommendations.

This Consensus Statement provides recommendations on the prescription of pain medication at discharge from hospital for opioid-naïve adult patients who undergo elective surgery. It encourages health care providers (surgeons, anesthesiologists, nurses/nurse practitioners, pain teams, pharmacists, allied health professionals, and trainees) to (1) use nonopioid therapies and reduce the prescription of opioids so that fewer opioid pills are available for diversion and (2) educate patients and their families/caregivers about pain management options after surgery to optimize quality of care for postoperative pain. These recommendations apply to opioid-naïve adult patients who undergo elective surgery. This consensus statement is intended for use by health care providers involved in the management and care of surgical patients. A modified Delphi process was used to reach consensus on the recommendations. First, the authors conducted a scoping review of the literature to determine current best practices and existing guidelines. From the available literature and expertise of the authors, a draft list of recommendations was created. Second, the authors asked key stakeholders to review and provide feedback on several drafts of the document and attend an in-person consensus meeting. The modified Delphi stakeholder group included surgeons, anesthesiologists, residents, fellows, nurses, pharmacists, and patients. After multiple iterations, the document was deemed complete. The recommendations are not graded because they are mostly based on consensus rather than evidence., Competing Interests: No potential conflict of interest was reported by the authors., (© 2020 The Author(s). Published with license by Taylor & Francis Group, LLC.)

Background: The Duke Activity Status Index (DASI) questionnaire might help incorporate self-reported functional capacity into preoperative risk assessment. Nonetheless, prognostically important thresholds in DASI scores remain unclear. We conducted a nested cohort analysis of the Measurement of Exercise Tolerance before Surgery (METS) study to characterise the association of preoperative DASI scores with postoperative death or complications., Methods: The analysis included 1546 participants (≥40 yr of age) at an elevated cardiac risk who had inpatient noncardiac surgery. The primary outcome was 30-day death or myocardial injury. The secondary outcomes were 30-day death or myocardial infarction, in-hospital moderate-to-severe complications, and 1 yr death or new disability. Multivariable logistic regression modelling was used to characterise the adjusted association of preoperative DASI scores with outcomes., Results: The DASI score had non-linear associations with outcomes. Self-reported functional capacity better than a DASI score of 34 was associated with reduced odds of 30-day death or myocardial injury (odds ratio: 0.97 per 1 point increase above 34; 95% confidence interval [CI]: 0.96-0.99) and 1 yr death or new disability (odds ratio: 0.96 per 1 point increase above 34; 95% CI: 0.92-0.99). Self-reported functional capacity worse than a DASI score of 34 was associated with increased odds of 30-day death or myocardial infarction (odds ratio: 1.05 per 1 point decrease below 34; 95% CI: 1.00-1.09), and moderate-to-severe complications (odds ratio: 1.03 per 1 point decrease below 34; 95% CI: 1.01-1.05)., Conclusions: A DASI score of 34 represents a threshold for identifying patients at risk for myocardial injury, myocardial infarction, moderate-to-severe complications, and new disability., (Copyright © 2019 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Objective: The aim of this study was to summarize strategies to reduce postsurgical opioid prescribing at discharge., Summary Background Data: Current practices for the prescription of opioids at discharge after surgery are highly variable and often excessive. We conducted a systematic review to identify behavioral interventions designed to improve these practices., Methods: We searched MEDLINE, EMBASE, CINAHL, and PsycINFO until December 14, 2018 to identify studies of behavioral interventions designed to decrease opioid prescribing at discharge among adults undergoing surgery. Behavioral interventions were defined according to the Cochrane Effective Practice and Organisation of Care (EPOC) taxonomy. We assessed the risk of bias of included studies using criteria suggested by Cochrane EPOC and the Newcastle-Ottawa scale., Results: Of 8048 citations that were screened, 24 studies were included in our review. Six types of behavioral interventions were identified: local consensus-based processes (18 studies), patient-mediated interventions (2 studies), clinical practice guidelines (1 study), educational meetings (1 study), interprofessional education (1 study), and clinician reminder (1 study). All but one study reported a statistically significant decrease in the amount of opioid prescribed at discharge after surgery, and only 2 studies reported evidence of increased pain intensity. Reductions in prescribed opioids ranged from 34.4 to 212.3 mg morphine equivalents. All studies were found to have medium-to-high risks of bias., Conclusions: We identified 6 types of behavioral strategies to decrease opioid prescription at discharge after surgery. Despite the risk of bias, almost all types of intervention seemed effective in reducing opioid prescriptions at discharge after surgery without negatively impacting pain control.

Introduction: The present study investigated the associations between smoking, pain, and opioid consumption in the 3 months after major surgery in patients seen by the Transitional Pain Service. Current smoking status and lifetime pack-years were expected to be related to higher pain intensity, more opioid use, and poorer opioid weaning after surgery., Methods: A total of 239 patients reported smoking status in their presurgical assessment (62 smokers, 92 past smokers, and 85 never smokers). Pain and daily opioid use were assessed in hospital before postsurgical discharge, at first outpatient visit (median of 1 month postsurgery), and at last outpatient visit (median of 3 months postsurgery). Pain was measured using numeric rating scale. Morphine equivalent daily opioid doses were calculated for each patient., Results: Current smokers reported significantly higher pain intensity (p < .05) at 1 month postsurgery than never smokers and past smokers. Decline in opioid consumption differed significantly by smoking status, with both current and past smokers reporting a less than expected decline in daily opioid consumption (p < .05) at 3 months. Decline in opioid consumption was also related to pack-years, with those reporting higher pack-years having a less than expected decline in daily opioid consumption at 3 months (p < .05)., Conclusions: Smoking status may be an important modifiable risk factor for pain intensity and opioid use after surgery., Implications: In a population with complex postsurgical pain, smoking was associated with greater pain intensity at 1 month after major surgery and less opioid weaning 3 months after surgery. Smoking may be an important modifiable risk factor for pain intensity and opioid use after surgery.

Introduction: Cannabis sativa (hemp) seeds are popular for their high nutrient content, and strict regulations are in place to limit the amount of potentially harmful phytocannabinoids, especially Δ 9 -tetrahydrocannabinol (Δ 9 -THC). In Canada, this limit is 10 μg of Δ 9 -THC per gram of hemp seeds (10 ppm), and other jurisdictions in the world follow similar guidelines. Materials and Methods: We investigated three different brands of consumer-grade hemp seeds using four different procedures to extract phytocannabinoids, and quantified total Δ 9 -THC and cannabidiol (CBD). Discussion: We discovered that Δ 9 -THC concentrations in these hemp seeds could be as high as 1250% of the legal limit, and the amount of phytocannabinoids depended on the extraction procedure employed, Soxhlet extraction being the most efficient across all three brands of seeds. Δ 9 -THC and CBD exhibited significant variations in their estimated concentrations even from the same brand, reflecting the inhomogeneous nature of seeds and variability due to the extraction method, but almost in all cases, Δ 9 -THC concentrations were higher than the legal limit. These quantities of total Δ 9 -THC may reach as high as 3.8 mg per gram of hemp seeds, if one were consuming a 30-g daily recommended amount of hemp seeds, and is a cause for concern for potential toxicity. It is not clear if these high quantities of Δ 9 -THC are due to contamination of the seeds, or any other reason. Conclusion: Careful consideration of the extraction method is very important for the measurement of cannabinoids in hemp seeds., Competing Interests: L.P.K. and H.A.C. serve on the scientific and medical advisory board of Scientus Pharma, Inc. and receive a consulting fee.

Medical cannabis has been legally available for patients in a number of countries. Licensed producers produce a variety of cannabis strains with different concentrations of phytocannabinoids. Phytocannabinoids in medical cannabis are decarboxylated when subjected to heating for consumption by the patients or when extracted for preparing cannabis derivative products. There is little understanding of the true chemical composition of cannabis extracts, changes occurring during heating of the extracts, and their relevance to pharmacological effects. We investigated the extract from a popular commercial strain of medical cannabis, prior to and after decarboxylation, to understand the chemical profiles. A total of up to 62 compounds could be identified simultaneously in the extract derived from commercial cannabis, including up to 23 phytocannabinoids. Upon heating, several chemical changes take place, including the loss of carboxylic group from the acidic phytocannabinoids. This investigation attempts to reveal the chemical complexity of commercial medical cannabis extracts and the differences in the chemical composition of the native extract and the one subjected to heat. Comprehensive chemical analyses of medical cannabis extracts are needed for standardization, consistency, and, more importantly, an informed employment of this substance for therapeutic purposes., Competing Interests: The authors declare the following competing financial interest(s): L.P.K. and H.A.C. serve on the scientific and medical advisory board of Scientus Pharma, Inc. and receive a consulting fee.

Purpose: The aim of this study was to explore acute movement-evoked postoperative pain intensity trajectories over the first 5 days after total hip arthroplasty (THA) and to examine how these pain trajectories are associated with pain-related outcomes 6 weeks and 6 months later., Methods: A total of 150 adult patients [72 women (48.0 %); mean age 60.0 ± 9.2 (standard deviation) years] completed pain questionnaires preoperatively, several times daily postoperatively until hospital discharge, and 6 weeks and 6 months after surgery., Results: Results showed that the best model had four different acute postoperative pain trajectories and a significant quadratic term. The trajectories varied in terms of initial pain intensity levels and rates of decline/increase in pain over the first 4 postoperative days. Significant predictors of pain trajectory membership were preoperative pain disability and anxiety as well as cumulative morphine consumption 24 h following surgery. Pain trajectories were significantly associated with levels of pain intensity and anxiety at 6 weeks but not at 6 months postoperatively., Conclusion: This study showed that during the postoperative period patients differed in terms of pain intensity profiles and that these differences were associated with outcomes for up to 6 weeks following surgery. Pain trajectories were not predictive of persistent postoperative pain status at 6 months. Nonetheless, these results highlight the importance of patient heterogeneity in acute postoperative pain and pain-related outcomes months after THA.

Background and Objective: Hyperbaric oxygen therapy (HBOT) is a treatment providing 100% oxygen at a pressure greater than that at sea level. HBOT is becoming increasingly recognized as a potential treatment modality for a broad range of ailments, including chronic pain. In this narrative review, we discuss the current understanding of pathophysiology of nociceptive, inflammatory and neuropathic pain, and the body of animal studies addressing mechanisms by which HBOT may ameliorate these different types of pain. Finally, we review clinical studies suggesting that HBOT may be useful in treating chronic pain syndromes, including chronic headache, fibromyalgia, complex regional pain syndrome, and trigeminal neuralgia., Database and Data Treatment: A comprehensive search through MEDLINE, EMBASE, Scopus, and Web of Science for studies relating to HBOT and pain was performed using the following keywords: hyperbaric oxygen therapy or hyperbaric oxygen treatment (HBOT), nociceptive pain, inflammatory pain, neuropathic pain, HBOT AND pain, HBOT AND headache, HBOT AND fibromyalgia, HBOT AND complex regional pain syndrome, and HBOT AND trigeminal neuralgia., Results: Twenty-five studies examining the role of HBOT in animal models of pain and human clinical trials were found and reviewed for this narrative review., Conclusions: HBOT has been shown to reduce pain using animal models. Early clinical research indicates HBOT may also be useful in modulating human pain; however, further studies are required to determine whether HBOT is a safe and efficacious treatment modality for chronic pain conditions., (© 2015 World Institute of Pain.)

The goal of this study was to follow a cohort of patients undergoing total knee arthroplasty over time to: (1) identify and describe the various pain trajectories beginning preoperatively and for up to 12 months after surgery, (2) identify baseline predictors of trajectory group membership, and (3) identify trajectory groups associated with poor psychosocial outcomes 12 months after surgery. One hundred seventy-three participants (female = 85 [49%]; mean age [years] = 62.9, SD = 6.8) completed pain and psychological questionnaires and functional performance tests preoperatively and 4 days, 6 weeks, and 3 and 12 months after total knee arthroplasty. Using growth mixture modeling, results showed that a 4-group model, with a quadratic slope and baseline pain data predicting trajectory group membership, best fit the data (Akaike information criterion = 2772.27). The first 3 pain trajectories represent various rates of recovery ending with relatively low levels of pain 12 months after surgery. Group 4, the constant high pain group, comprises patients who have a neutral or positive pain slope and do not show improvement in their pain experience over the first year after surgery. This model suggests that preoperative pain levels are predictive of pain trajectory group membership and moderate preoperative pain, as opposed to low or high pain, is a risk factor for a neutral or positive pain trajectory postoperatively. Consistent with previous studies, these results show that postoperative pain is not a homogeneous condition and point to the importance of examining intraindividual pain fluctuations as they relate to pain interventions and prevention strategies.

Background: This study was designed to determine whether a 4 day perioperative regimen of gabapentin added to celecoxib improves in-hospital rehabilitation and physical function on postoperative day 4 and 6 weeks and 3 months after total knee arthroplasty (TKA)., Methods: After Research Ethics Board approval and informed consent, 212 patients were enrolled in a randomized, double-blinded, placebo-controlled study. Two hours before surgery, patients received celecoxib 400 mg p.o. and were randomly assigned to receive either gabapentin 600 mg or placebo p.o. Two hours later, patients received femoral, sciatic nerve blocks, and spinal anaesthesia. After operation, patients received gabapentin 200 mg or placebo three times per day (TID) for 4 days. All patients also received celecoxib 200 mg q12 h for 72 h and i.v. patient-controlled analgesia for 24 h. Pain and function were assessed at baseline, during hospitalization, on postoperative day 4 (POD4), and 6 weeks and 3 months after surgery., Results: The gabapentin group used less morphine in the first 24 h after surgery [G=38.3 (29.5 mg), P=48.2 (29.4 mg)] (P<0.0125) and had increased knee range of motion compared with the placebo group in-hospital (P<0.05). There were no differences between groups in favour of the gabapentin group for pain or physical function on POD 4 [95% confidence interval (CI): pain: -1.4, 0.5; function: -6.3, 2.0], 6 weeks (95% CI: pain: 0.1, 1.9; function: -0.2, 6.5) or 3 months (95% CI: pain: -0.2, 1.7; function: -2.2, 4.3) after TKA., Conclusions: In the context of celecoxib, spinal anaesthesia, femoral and sciatic nerve blocks, a dose of gabapentin 600 mg before operation followed by 4 days of gabapentin 200 mg TID decreased postoperative analgesic requirements and improved knee range of motion after TKA. Gabapentin provided no improvement in pain or physical function on POD4 and 6 weeks or 3 months after surgery., (© The Author 2014. Published by Oxford University Press on behalf of the British Journal of Anaesthesia. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Despite its prominence as a concern among potential surgical candidates, there is little information in the literature regarding the short- and long-term pain experience after living liver donation. We undertook a prospective study to examine (1) the nature and incidence of acute and chronic pain after living donor hepatectomy and (2) the factors associated with an increased or decreased risk of adverse pain outcomes. Before donation, a comprehensive assessment of potential predictors of acute and chronic pain outcomes was conducted; this included donors' pain expectations, psychosocial factors, medical histories, and demographic factors. Detailed data regarding pain outcomes were collected postoperatively (days 1 and 2) and again during 6- and 12-month follow-up telephone interviews. Sixty-five adults (32 females and 33 males) scheduled for donor hepatectomy participated. Substantial proportions of the donors reported a moderate-to-severe level of pain intensity (≥4 on a 0-10 scale) at rest and after movement on day 1 (42% and 74%, respectively) and day 2 (33% and 32%, respectively). Persistent postsurgical pain was reported by 31% of the donors at the 6-month follow-up and by 27% of the donors at the 12-month follow-up. Generally, this pain was mild, and pain-related life interference was minimal. Female sex, a younger age, and several predonation measures of pain-related anxiety were associated with a significantly greater risk of developing persistent postsurgical pain. In conclusion, this study has identified a subset of patients who experience persistent pain after living liver donation. Additional prospective research using larger samples of liver donors is needed to replicate this work, to obtain a more detailed account of the acute and long-term pain experience, and to determine whether targeted interventions can minimize the frequency and severity of chronic pain., (© 2014 American Association for the Study of Liver Diseases.)

Purpose: Antidepressant medications are commonly prescribed for the treatment of depression, anxiety, and chronic pain. Their use may lead to a number of side effects with important implications in the perioperative period. Our aim was to examine the effect of preoperative antidepressant administration on post-surgical hospital length of stay (LOS) in elective non-cardiac surgery patients., Design: Historical cohort study., Methods: Demographic and preoperative data were collected by chart review for all non-cardiac surgery patients who were assessed in the preoperative consult clinic from April 2008 through February 2009. Patients were grouped according to whether or not they were taking antidepressant medications. Median length of stay was compared between patients who took antidepressants preoperatively and those who did not., Results: Data were collected for 3,692 patients. Two hundred eighty-nine (7.8%) patients were taking antidepressants preoperatively. Use of antidepressants was not associated with an increased hospital LOS. The median LOS was four days both for patients who took antidepressants preoperatively (95% confidence interval [CI] 4 to 4) and for those who did not (95% CI 3 to 5) (P = 0.13)., Conclusions: The preoperative use of antidepressant medications was not associated with increased postoperative hospital LOS following elective non-cardiac surgery.

Purpose: To determine effective spinal anesthetic dosing of hyperbaric bupivacaine for patients undergoing high-dose-rate (HDR) brachytherapy for prostate cancer., Methods: A total of 199 HDR prostate brachytherapy sessions (n=115 patients) under spinal anesthesia with five doses of hyperbaric bupivacaine were prospectively analyzed. Patients received either 11.25mg (n=40 sessions), 15mg (n=63 sessions), 18mg (n=25 sessions), 18.75mg (n=17 sessions), or 20.25mg (n=54 sessions) of 0.75% of bupivacaine with 20microg of fentanyl in 8.25% of dextrose (Hospira, Healthcare Corporation, Montréal, Canada). Sensory block level, intraoperative hypotension, incidence of nausea and vomiting, time to ambulation, and time to discharge were recorded. We assessed for the presence of any transient neurologic symptoms post discharge., Results: The 11.25-mg group had a mean peak sensory block height of L1 at 90min, which was significantly lower than the other groups. Block heights were also decreased at the 5, 60, 120, 150, 180, and 300min time points (p<0.05). The mean time to ambulation was 403+/-92min and to discharge was 427+/-70min; no difference among groups was found. Hypotension was not observed in the 11.25-mg group, whereas the incidence of operative hypotension was 8% in each of the other groups. No transient neurologic symptoms were reported postoperatively., Conclusions: The 11.25-mg dose of hyperbaric bupivacaine was sufficient to provide intraoperative anesthesia for catheter placement and sufficient postoperative residual analgesia for the duration that the catheters were left in situ and subsequently removed.

Background: The aim of this study was to investigate the influence of current hormonal contraceptive medication on periodontal health in young females., Methods: Fifty women aged 20 to 35 years (mean +/- SD: 29.7 +/- 4.7 years) had a comprehensive periodontal examination. Current and previous contraceptive pill use was assessed by a questionnaire. Periodontal assessment included plaque index, gingival index, probing depth, and attachment level at six sites per tooth. The periodontal health of current pill users was compared to that of women not taking the pill., Results: Forty-two percent of subjects were taking the contraceptive pill at the time of periodontal examination. Current pill users had deeper mean probing depths compared to non-users (3.3 mm versus 2.7 mm; P = 0.006) and more severe attachment loss (2.6 mm versus 1.7 mm; P = 0.015). Pill users had more sites with bleeding on probing (44.0% versus 31.1%; P = 0.017)., Conclusion: Current users of oral contraceptives had poorer periodontal health.

This study compared rats with dorsal striatal, ventrolateral prefrontal cortical, and combined lesions of the hippocampus and amygdala to sham controls on a conditional discrimination task in which contextual cues modulated a taste aversion. All groups were able to acquire this occasion setting task. The 2nd experiment functionally minimized the stimulus-response component of the paradigm, creating a "tasteless" form of occasion setting. Rats with pretraining lesions of the hippocampus and amygdala were impaired compared with shams on the acquisition of this tasteless occasion setting task. Rats with posttraining combined lesions did not retain the ability to perform the tasteless occasion setting task learned preoperatively. Rats with selective lesions of either the hippocampus or the amygdala alone were not impaired in the acquisition of the tasteless occasion setting task. The findings suggest that this occasion setting task may be learned by several redundant neural systems.

Although indirect evidence suggests that the control of sympathetic preganglionic neurons is mediated to a great extent through interneurons, little is known about the location, morphology or neurotransmitter phenotype of such interneurons. This limitation seriously impedes our understanding of spinal synaptic circuits crucial to control of arterial pressure and other visceral functions. We used a highly neurotropic, minimally cytopathic recombinant herpes simplex virus type-1 to study spinal "sympathetic" interneurons labelled by trans-synaptic transport of the virus from the adrenal gland in rats. Approximately 120-320 infected neurons/rat were identified by immunocytochemical detection of the viral antigen. We distinguished between virus-infected preganglionic neurons and infected interneurons by (i) their location within the spinal laminae, (ii) their size and shape and (iii) the presence or absence of immunoreactivity for the acetylcholine-synthesizing enzyme, choline acetyltransferase, a marker of sympathetic preganglionic neurons. Virus-labelled sympathetic preganglionic neurons were found within the known spinal preganglionic nuclei. Non-cholinergic, virus-labelled neurons were located throughout lamina VII and in the ventral portion of lamina V. These putative interneurons were found in the major spinal preganglionic nuclei, usually intermingled with the preganglionic neurons. Sometimes, they were located in clusters separate from the preganglionic neurons. The interneurons were approximately 15 microm in diameter, smaller than the average preganglionic neuron (diameter=25 microm), and had a few fine processes emanating from them. These non-cholinergic interneurons constituted approximately one-half of the population of virus-infected neurons. In summary, with the use of a recombinant herpes simplex virus, we identified a large number of non-cholinergic interneurons close to, or intermingled with, adrenal sympathetic preganglionic neurons. The neurotransmitter phenotype of these neurons remains to be determined but they likely integrate much of the supraspinal and primary afferent inputs to spinal preganglionic neurons that control arterial pressure and other visceral functions.

A case of a 36-year-old woman with a past history of uterine leiomyosarcoma and an intracerebral metastasis is reported. The patient presented with a 24-hour history of severe headache with coma, and the CT findings were consistent with a metastatic lesion. Pathological examination of the operative specimen showed features of a leiomyosarcoma. Uterine leiomyosarcoma is an uncommon tumor and metastasis to the brain is rare.

One mechanism by which viral tropism may be controlled is by the expression of a specific virus receptor on the cell surface. This paper reports the identification of a putative cellular receptor for visna virus, the prototype virus of the family Lentiviridae. Using a virus overlay protein blot assay we identified a group of polypeptides of apparent Mr 30K to 33K which interacts with visna virus and is present on permissive but not non-permissive cells. A rat polyclonal anti-ovine major histocompatibility complex (MHC) class II antigen (Ag) serum raised to immunopurified MHC class II Ag, but not preimmune serum, blocked the interaction of visna virus with these polypeptides. In an ELISA, immunopurified MHC class II Ag bound to visna virus but not to bovine parainfluenza 3 virus. Preincubation of visna virus with immunopurified soluble MHC class II Ag resulted in a marked decrease in virus-induced syncytium formation, i.e. preincubation with class II Ag inhibited infection with visna virus, but we have been unable to inhibit infection using class II Ag-specific antisera. These results suggest that ovine MHC class II Ag acts as a component of a cellular receptor for visna virus. This is of particular interest owing to the close similarities between visna virus and human immunodeficiency virus (HIV), and the relationship between MHC class II and CD4, the cellular receptor for HIV. It is also of relevance to recent reports that a growing number of viruses utilize polypeptides of the Ig supergene family as receptors.