116 results on '"Cunningham AM"'
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2. Introducing a multi-site program for early diagnosis of HIV infection among HIV-exposed infants in Tanzania
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Malisa Isaya, Nankabirwa Victoria, Mtunga Sevestine, Gamaliel John G, Cunningham Amy, Abdallah Aziz, Werq-Semo Bazghina, Nuwagaba-Biribonwoha Harriet, Gonzalez Luis F, Massambu Charles, Nash Denis, Justman Jessica, and Abrams Elaine J
- Subjects
Pediatrics ,RJ1-570 - Abstract
Abstract Background In Tanzania, less than a third of HIV infected children estimated to be in need of antiretroviral therapy (ART) are receiving it. In this setting where other infections and malnutrition mimic signs and symptoms of AIDS, early diagnosis of HIV among HIV-exposed infants without specialized virologic testing can be a complex process. We aimed to introduce an Early Infant Diagnosis (EID) pilot program using HIV DNA Polymerase Chain Reaction (PCR) testing with the intent of making EID nationally available based on lessons learned in the first 6 months of implementation. Methods In September 2006, a molecular biology laboratory at Bugando Medical Center was established in order to perform HIV DNA PCR testing using Dried Blood Spots (DBS). Ninety- six health workers from 4 health facilities were trained in the identification and care of HIV-exposed infants, HIV testing algorithms and collection of DBS samples. Paper-based tracking systems for monitoring the program that fed into a simple electronic database were introduced at the sites and in the laboratory. Time from birth to first HIV DNA PCR testing and to receipt of test results were assessed using Kaplan-Meier curves. Results From October 2006 to March 2007, 510 HIV-exposed infants were identified from the 4 health facilities. Of these, 441(87%) infants had an HIV DNA PCR test at a median age of 4 months (IQR 1 to 8 months) and 75(17%) were PCR positive. Parents/guardians for a total of 242(55%) HIV-exposed infants returned to receive PCR test results, including 51/75 (68%) of those PCR positive, 187/361 (52%) of the PCR negative, and 4/5 (80%) of those with indeterminate PCR results. The median time between blood draw for PCR testing and receipt of test results by the parent or guardian was 5 weeks (range Conclusions The EID pilot program successfully introduced systems for identification of HIV-exposed infants. There was a high response as hundreds of HIV-exposed infants were registered and tested in a 6 month period. Challenges included the large proportion of parents not returning for PCR test results. Experience from the pilot phase has informed the national roll-out of the EID program currently underway in Tanzania.
- Published
- 2010
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3. Serotonin Transporter-dependent Histone Serotonylation in Placenta Contributes to the Neurodevelopmental Transcriptome.
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Chan JC, Alenina N, Cunningham AM, Ramakrishnan A, Shen L, Bader M, and Maze I
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- Animals, Female, Mice, Pregnancy, Mice, Transgenic, Transcriptome, Histones metabolism, Placenta metabolism, Serotonin metabolism, Serotonin Plasma Membrane Transport Proteins genetics, Serotonin Plasma Membrane Transport Proteins metabolism, Brain embryology, Receptors, Serotonin genetics, Receptors, Serotonin metabolism, Gene Expression Regulation, Developmental, Neurogenesis genetics
- Abstract
Brain development requires appropriate regulation of serotonin (5-HT) signaling from distinct tissue sources across embryogenesis. At the maternal-fetal interface, the placenta is thought to be an important contributor of offspring brain 5-HT and is critical to overall fetal health. Yet, how placental 5-HT is acquired, and the mechanisms through which 5-HT influences placental functions, are not well understood. Recently, our group identified a novel epigenetic role for 5-HT, in which 5-HT can be added to histone proteins to regulate transcription, a process called H3 serotonylation. Here, we show that H3 serotonylation undergoes dynamic regulation during placental development, corresponding to gene expression changes that are known to influence key metabolic processes. Using transgenic mice, we demonstrate that placental H3 serotonylation is dependent on 5-HT uptake by the serotonin transporter (SERT/SLC6A4). SERT deletion robustly reduces enrichment of H3 serotonylation across the placental genome, and disrupts neurodevelopmental gene networks in early embryonic brain tissues. Thus, these findings suggest a novel role for H3 serotonylation in coordinating placental transcription at the intersection of maternal physiology and offspring brain development., 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 © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2024
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4. Pathologic Features of Primary Pancreatic Malignancies.
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Abi-Saab T, Cunningham AM, Rush PS, and Matkowskyj KA
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- Humans, Neuroendocrine Tumors pathology, Pancreatic Neoplasms pathology
- Abstract
This chapter explores the pathologic features of benign and malignant lesions of the pancreas. As pathologic classifications evolve particularly for cystic lesions and neuroendocrine tumors, it is important for physicians who treat patients with gastrointestinal malignance to fully evaluate these pathologic classifications., (© 2024. The Author(s), under exclusive license to Springer Nature Switzerland AG.)
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- 2024
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5. Serotonin transporter-dependent histone serotonylation in placenta contributes to the neurodevelopmental transcriptome.
- Author
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Chan JC, Alenina N, Cunningham AM, Ramakrishnan A, Shen L, Bader M, and Maze I
- Abstract
Brain development requires appropriate regulation of serotonin (5-HT) signaling from distinct tissue sources across embryogenesis. At the maternal-fetal interface, the placenta is thought to be an important contributor of offspring brain 5-HT and is critical to overall fetal health. Yet, how placental 5-HT is acquired, and the mechanisms through which 5-HT influences placental functions, are not well understood. Recently, our group identified a novel epigenetic role for 5-HT, in which 5-HT can be added to histone proteins to regulate transcription, a process called H3 serotonylation. Here, we show that H3 serotonylation undergoes dynamic regulation during placental development, corresponding to gene expression changes that are known to influence key metabolic processes. Using transgenic mice, we demonstrate that placental H3 serotonylation largely depends on 5-HT uptake by the serotonin transporter (SERT/SLC6A4). SERT deletion robustly reduces enrichment of H3 serotonylation across the placental genome, and disrupts neurodevelopmental gene networks in early embryonic brain tissues. Thus, these findings suggest a novel role for H3 serotonylation in coordinating placental transcription at the intersection of maternal physiology and offspring brain development., Competing Interests: Declaration of Interest The authors declare no competing interests.
- Published
- 2023
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6. Patent ductus arteriosus (PDA) and response to late surfactant treatment in premature infants.
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Beauchene MS, Cunningham AM, Stanford AH, Bischoff AR, Dagle JM, Rios DR, Klein JM, Giesinger RE, and McNamara PJ
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Objective: To determine the clinical/echocardiography (ECHO) phenotype of patients with hypoxic respiratory failure (HRF) and response to late surfactant, according to patent ductus arteriosus (PDA) status., Study Design: This retrospective study included infants ≤26
+6 weeks gestation who received ≥1 surfactant dose after 6 postnatal days and where PDA status was available by ECHO. Response to surfactant was appraised based on change in respiratory severity score over 48 h. The relationship between PDA status and response to surfactant was evaluated via univariate analysis., Result: We studied late surfactant (n = 71 doses) administration in 35 preterm infants born at a mean weight and GA at birth were 595 g (508, 696) and 23.3 (22.7, 25) weeks, respectively of whom 16 (46%) had a diagnosis of PDA. Positive response to late surfactant treatment was independently associated with absence of PDA [OR 26 (2, 334), p = 0.01] whereas presence of PDA was independently associated with negative response [OR 12 (1.1, 126), p = 0.04]., Conclusions: In neonates ≤26+6 weeks gestation, with HRF, response to surfactant after postnatal day 6 is influenced by PDA status. Future trials should consider PDA status which may enhance diagnostic precision and refine patient selection for late surfactant treatment., (© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)- Published
- 2023
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7. Adhesive Wearable Sensors for Electroencephalography from Hairy Scalp.
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Zhang A, Shyam AB, Cunningham AM, Williams C, Brissenden A, Bartley A, Amsden B, Docoslis A, Kontopoulou M, and Ameri SK
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- Humans, Adhesives, Skin, Electroencephalography, Electrodes, Scalp, Wearable Electronic Devices
- Abstract
Electroencephalography has garnered interest for applications in mobile healthcare, human-machine interfaces, and Internet of Things. Conventional electroencephalography relies on wet and dry electrodes. Despite favorable interface impedance of wet electrodes and skin, the application of a large amount of gel at their interface with skin limits the electroencephalography spatial resolution, increases the risk of shorting between electrodes, and makes them unsuited for long-term mobile recording. In contrast, dry electrodes are better suited for long-term recordings but susceptible to motion artifacts. In addition, both wet and dry electrodes are non-adhesive to the hairy scalp and mechanical support, or chemical adhesives are used to hold them in place. Herein, a conical microstructure array (CMSA) based sensor made of carbon nanotube-polydimethylsiloxane composite is reported. The CMSA sensor is fabricated using the innovative, cost-effective, and scalable method of viscosity-controlled dip-pull process. The sensor adheres to the hairy scalp by generating negative pressure in its conical microstructures when it is pressed against scalp. Aided by the application of a trace amount of gel, CMSA sensor establishes good electrical contact with the skin, enabling its applications in mobile electroencephalography over extended periods. Notably, the signal quality of CMSA sensors is comparable to that of medical-grade wet gel electrodes., (© 2023 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.)
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- 2023
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8. Transcriptional signatures of heroin intake and relapse throughout the brain reward circuitry in male mice.
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Browne CJ, Futamura R, Minier-Toribio A, Hicks EM, Ramakrishnan A, Martínez-Rivera FJ, Estill M, Godino A, Parise EM, Torres-Berrío A, Cunningham AM, Hamilton PJ, Walker DM, Huckins LM, Hurd YL, Shen L, and Nestler EJ
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- Humans, Mice, Male, Animals, Genome-Wide Association Study, Brain, Reward, Recurrence, Heroin adverse effects, Opioid-Related Disorders
- Abstract
Opioid use disorder (OUD) looms as one of the most severe medical crises facing society. More effective therapeutics will require a deeper understanding of molecular changes supporting drug-taking and relapse. Here, we develop a brain reward circuit-wide atlas of opioid-induced transcriptional regulation by combining RNA sequencing (RNA-seq) and heroin self-administration in male mice modeling multiple OUD-relevant conditions: acute heroin exposure, chronic heroin intake, context-induced drug-seeking following abstinence, and relapse. Bioinformatics analysis of this rich dataset identified numerous patterns of transcriptional regulation, with both region-specific and pan-circuit biological domains affected by heroin. Integration of RNA-seq data with OUD-relevant behavioral outcomes uncovered region-specific molecular changes and biological processes that predispose to OUD vulnerability. Comparisons with human OUD RNA-seq and genome-wide association study data revealed convergent molecular abnormalities and gene candidates with high therapeutic potential. These studies outline molecular reprogramming underlying OUD and provide a foundational resource for future investigations into mechanisms and treatment strategies.
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- 2023
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9. Pediatric Myeloid Sarcoma, More than Just a Chloroma: A Review of Clinical Presentations, Significance, and Biology.
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Zorn KE, Cunningham AM, Meyer AE, Carlson KS, and Rao S
- Abstract
Myeloid sarcomas (MS), commonly referred to as chloromas, are extramedullary tumors of acute myeloid leukemia (AML) with varying incidence and influence on outcomes. Pediatric MS has both a higher incidence and unique clinical presentation, cytogenetic profile, and set of risk factors compared to adult patients. Optimal treatment remains undefined, yet allogeneic hematopoietic stem cell transplantation (allo-HSCT) and epigenetic reprogramming in children are potential therapies. Importantly, the biology of MS development is poorly understood; however, cell-cell interactions, epigenetic dysregulation, cytokine signaling, and angiogenesis all appear to play key roles. This review describes pediatric-specific MS literature and the current state of knowledge about the biological determinants that drive MS development. While the significance of MS remains controversial, the pediatric experience provides an opportunity to investigate mechanisms of disease development to improve patient outcomes. This brings the hope of better understanding MS as a distinct disease entity deserving directed therapeutic approaches.
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- 2023
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10. Synthesis and Stereochemical Determination of the Peptide Antibiotic Novo29.
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Krumberger M, Li X, Kreutzer AG, Peoples AJ, Nitti AG, Cunningham AM, Jones CR, Achorn C, Ling LL, Hughes DE, and Nowick JS
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- Models, Molecular, Molecular Conformation, Magnetic Resonance Spectroscopy, Anti-Bacterial Agents pharmacology, Anti-Bacterial Agents chemistry, Amino Acids chemistry
- Abstract
This paper describes the synthesis and stereochemical determination of Novo29 (clovibactin), a new peptide antibiotic that is related to teixobactin and is active against Gram-positive bacteria. Novo29 is an eight-residue depsipeptide that contains the noncanonical amino acid hydroxyasparagine of hitherto undetermined stereochemistry in a macrolactone ring. The amino acid building blocks Fmoc-(2 R ,3 R )-hydroxyasparagine-OH and Fmoc-(2 R ,3 S )-hydroxyasparagine-OH were synthesized from ( R , R )- and ( S , S )-diethyl tartrate. Novo29 and epi -Novo29 were then prepared by solid-phase peptide synthesis using these building blocks. Correlation with an authentic sample of Novo29 through
1 H NMR spectroscopy, LC-MS, and in vitro antibiotic activity established that Novo29 contains (2 R ,3 R )-hydroxyasparagine. X-ray crystallography reveals that epi -Novo29 adopts an amphiphilic conformation, with a hydrophobic surface and a hydrophilic surface. Four sets of epi -Novo29 molecules pack in the crystal lattice to form a hydrophobic core. The macrolactone ring adopts a conformation in which the main-chain amide NH groups converge to create a cavity, which binds ordered water and acetate anion. The amphiphilic conformation of epi -Novo29 is reminiscent of the amphiphilic conformation adopted by the related antibiotic teixobactin and its derivatives, which contains a hydrophobic surface that interacts with the lipids of the bacterial cell membrane and a hydrophilic surface that interacts with the aqueous environment. Molecular modeling suggests that Novo29 can adopt an amphiphilic conformation similar to teixobactin, suggesting that Novo29 may interact with bacteria in a similar fashion to teixobactin.- Published
- 2023
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11. Convergent abnormalities in striatal gene networks in human cocaine use disorder and mouse cocaine administration models.
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Mews P, Cunningham AM, Scarpa J, Ramakrishnan A, Hicks EM, Bolnick S, Garamszegi S, Shen L, Mash DC, and Nestler EJ
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- Male, Humans, Mice, Animals, Gene Regulatory Networks, Receptors, Dopamine D2 genetics, Receptors, Dopamine D2 metabolism, Brain metabolism, Cocaine pharmacology, Cocaine-Related Disorders genetics
- Abstract
Cocaine use disorder (CUD) is an intractable syndrome, and rising overdose death rates represent a substantial public health crisis that exacts tremendous personal and financial costs on patients and society. Sharp increases in cocaine use drive the urgent need for better mechanistic insight into this chronic relapsing brain disorder that currently lacks effective treatment options. To investigate the transcriptomic changes involved, we conducted RNA sequencing on two striatal brain regions that are heavily implicated in CUD, the nucleus accumbens and caudate nucleus, from men suffering from CUD and matched controls. Weighted gene coexpression analyses identified CUD-specific gene networks enriched in ionotropic receptors and linked to lowered neuroinflammation, contrasting the proinflammatory responses found in opioid use disorder. Integration of comprehensive transcriptomic datasets from mouse cocaine self-administration models revealed evolutionarily conserved gene networks in CUD that implicate especially D1 medium spiny neurons as drivers of cocaine-induced plasticity.
- Published
- 2023
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12. Transcriptional signatures of heroin intake and seeking throughout the brain reward circuit.
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Browne CJ, Futamura R, Minier-Toribio A, Hicks EM, Ramakrishnan A, Martínez-Rivera F, Estill M, Godino A, Parise EM, Torres-Berrío A, Cunningham AM, Hamilton PJ, Walker DM, Huckins LM, Hurd YL, Shen L, and Nestler EJ
- Abstract
Opioid use disorder (OUD) looms as one of the most severe medical crises currently facing society. More effective therapeutics for OUD requires in-depth understanding of molecular changes supporting drug-taking and relapse. Recent efforts have helped advance these aims, but studies have been limited in number and scope. Here, we develop a brain reward circuit-wide atlas of opioid-induced transcriptional regulation by combining RNA sequencing (RNAseq) and heroin self-administration in male mice modeling multiple OUD-relevant conditions: acute heroin exposure, chronic heroin intake, context-induced drug-seeking following prolonged abstinence, and heroin-primed drug-seeking (i.e., "relapse"). Bioinformatics analysis of this rich dataset identified numerous patterns of molecular changes, transcriptional regulation, brain-region-specific involvement in various aspects of OUD, and both region-specific and pan-circuit biological domains affected by heroin. Integrating RNAseq data with behavioral outcomes using factor analysis to generate an "addiction index" uncovered novel roles for particular brain regions in promoting addiction-relevant behavior, and implicated multi-regional changes in affected genes and biological processes. Comparisons with RNAseq and genome-wide association studies from humans with OUD reveal convergent molecular regulation that are implicated in drug-taking and relapse, and point to novel gene candidates with high therapeutic potential for OUD. These results outline broad molecular reprogramming that may directly promote the development and maintenance of OUD, and provide a foundational resource to the field for future research into OUD mechanisms and treatment strategies.
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- 2023
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13. A Step toward Understanding the Story Behind the Pictures: Molecular Diagnostics and the Banff Classification of Renal Allograft Pathology.
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Bissonnette MLZ, Riazy M, Cunningham AM, and Gill JS
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- Pathology, Molecular, Kidney pathology, Transplantation, Homologous, Allografts, Kidney Transplantation
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- 2022
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14. Crystallin Mu in Medial Amygdala Mediates the Effect of Social Experience on Cocaine Seeking in Males but Not in Females.
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Walker DM, Zhou X, Cunningham AM, Ramakrishnan A, Cates HM, Lardner CK, Peña CJ, Bagot RC, Issler O, Van der Zee Y, Lipschultz AP, Godino A, Browne CJ, Hodes GE, Parise EM, Torres-Berrio A, Kennedy PJ, Shen L, Zhang B, and Nestler EJ
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- Animals, Male, Female, Mice, mu-Crystallins, Reward, Neurons metabolism, Amygdala metabolism, Cocaine pharmacology, Cocaine metabolism
- Abstract
Background: Social experiences influence susceptibility to substance use disorder. The adolescent period is associated with the development of social reward and is exceptionally sensitive to disruptions to reward-associated behaviors by social experiences. Social isolation (SI) during adolescence alters anxiety- and reward-related behaviors in adult males, but little is known about females. The medial amygdala (meA) is a likely candidate for the modulation of social influence on drug reward because it regulates social reward, develops during adolescence, and is sensitive to social stress. However, little is known regarding how the meA responds to drugs of abuse., Methods: We used adolescent SI coupled with RNA sequencing to better understand the molecular mechanisms underlying meA regulation of social influence on reward., Results: We show that SI in adolescence, a well-established preclinical model for addiction susceptibility, enhances preference for cocaine in male but not in female mice and alters cocaine-induced protein and transcriptional profiles within the adult meA particularly in males. To determine whether transcriptional mechanisms within the meA are important for these behavioral effects, we manipulated Crym expression, a sex-specific key driver gene identified through differential gene expression and coexpression network analyses, specifically in meA neurons. Overexpression of Crym, but not another key driver that did not meet our sex-specific criteria, recapitulated the behavioral and transcriptional effects of adolescent SI., Conclusions: These results show that the meA is essential for modulating the sex-specific effects of social experience on drug reward and establish Crym as a critical mediator of sex-specific behavioral and transcriptional plasticity., (Copyright © 2022 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)
- Published
- 2022
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15. A Population-Based Analysis of the Risk of Glomerular Disease Relapse after COVID-19 Vaccination.
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Canney M, Atiquzzaman M, Cunningham AM, Zheng Y, Er L, Hawken S, Zhao Y, and Barbour SJ
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- Adult, Humans, COVID-19 Vaccines adverse effects, Retrospective Studies, Cohort Studies, Recurrence, Chronic Disease, Vaccination, COVID-19 epidemiology, COVID-19 prevention & control, Glomerulonephritis, IGA pathology
- Abstract
Background: Although case reports have described relapses of glomerular disease after COVID-19 vaccination, evidence of a true association is lacking. In this population-level analysis, we sought to determine relative and absolute risks of glomerular disease relapse after COVID-19 vaccination., Methods: In this retrospective population-level cohort study, we used a centralized clinical and pathology registry (2000-2020) to identify 1105 adult patients in British Columbia, Canada, with biopsy-proven glomerular disease that was stable on December 14, 2020 (when COVID-19 vaccines first became available). The primary outcome was disease relapse, on the basis of changes in kidney function, proteinuria, or both. Vaccination was modeled as a 30-day time-varying exposure in extended Cox regression models, stratified on disease type., Results: During 281 days of follow-up, 134 (12.1%) patients experienced a relapse. Although a first vaccine dose was not associated with relapse risk (hazard ratio [HR]=0.67; 95% confidence interval [95% CI], 0.33 to 1.36), exposure to a second or third dose was associated with a two-fold risk of relapse (HR=2.23; 95% CI, 1.06 to 4.71). The pattern of relative risk was similar across glomerular diseases. The absolute increase in 30-day relapse risk associated with a second or third vaccine dose varied from 1%-2% in ANCA-related glomerulonephritis, minimal change disease, membranous nephropathy, or FSGS to 3%-5% in IgA nephropathy or lupus nephritis. Among 24 patients experiencing a vaccine-associated relapse, 4 (17%) had a change in immunosuppression, and none required a biopsy., Conclusions: In a population-level cohort of patients with glomerular disease, a second or third dose of COVID-19 vaccine was associated with higher relative risk but low absolute increased risk of relapse., (Copyright © 2022 by the American Society of Nephrology.)
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- 2022
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16. Use of Iatrogenic Lipid Emulsion and Subsequent Plasmapheresis for the Treatment of Amitriptyline Overdose.
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Laffin RC, Cunningham AM, Fitzgerald SA, Wiebe DA, Wells JA, Linzell JR, and Rose WN
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Plasmapheresis for the treatment of hypertriglyceridemia is relatively uncommon and mostly reported either in patients experiencing hypertriglyceridemia-induced acute pancreatitis or patients with therapy-resistant familial hypercholesterolemia. Standard therapies for hypertriglyceridemia include dietary modification and lipid-lowering medication. For severe hypertriglyceridemia, the risk of pancreatitis increases significantly as triglyceride levels increase above 1000 mg/dL, and current therapies are unable to reduce triglyceride levels rapidly enough. Here, we report a case of a 48-year-old male patient who presented to the emergency department due to an amitriptyline overdose. In addition to being started on IV sodium bicarbonate therapy, an intravenous 20% fat emulsion bolus at 1.5 mL/kg was administered followed by 0.25 mL/kg/min infusion for 4 hours as a strategy to absorb lipophilic amitriptyline. Two days posttreatment, he was noted to have substantial hypertriglyceridemia (serum triglycerides: 6,475 mg/dL). His amylase was within the normal range at 37 U/L (reference range: 20-100 U/L), his lipase was low at 40 U/L (reference range: 75-390 U/L), and he was without evidence of any clinical sequelae secondary to hypertriglyceridemia (e.g., pancreatitis). Due to the severity of his hypertriglyceridemia, plasmapheresis was initiated urgently for rapid reduction in serum triglyceride levels to prevent pancreatitis and end-organ damage. He underwent a 1-plasma volume exchange with 5% albumin as the replacement fluid. This reduced his triglyceride levels to 185 mg/dL (reference range: 3-149 mg/dL). His symptoms secondary to his amitriptyline overdose were also resolved. Here, we report a 2-step process of intravenous lipid emulsion followed by plasmapheresis for amitriptyline overdose., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2022 Ryan C. Laffin et al.)
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- 2022
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17. Unexplained Hematocrit Increase after Therapeutic Phlebotomy in a Patient with Marked Erythrocytosis.
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Machhi R, Cunningham AM, Hennrick K, Schaser KA, Williams EC, and Rose WN
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We report a patient with hereditary erythrocytosis who underwent a therapeutic phlebotomy and had a post-phlebotomy hematocrit that was higher than the pre-phlebotomy hematocrit. We could not discern a reason for this hematocrit increase after phlebotomy. Instead of performing another phlebotomy, we performed an automated red cell depletion via an apheresis instrument. This procedure is essentially a red cell exchange, but 5% albumin is used as the replacement fluid instead of red blood cells. The patient's hematocrit decreased from 80% to 39% after three consecutive daily red cell depletion procedures. We share our experience to report the unusual finding of a patient's hematocrit that increased with phlebotomy and to raise awareness of the red cell depletion procedure., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2022 Rushad Machhi et al.)
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- 2022
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18. Diffuse Leptomeningeal Histiocytic Sarcoma: Histologic and Molecular Findings in an Autopsy Case.
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Onyenekwu CP, Cunningham AM, Schilter K, Reddi HV, and Cochran EJ
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- Adult, Fatal Outcome, Female, Humans, Histiocytic Sarcoma pathology, Meningeal Carcinomatosis pathology
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- 2022
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19. Reply to: Multiple Comparisons and Inappropriate Statistical Testing Lead to Spurious Sex Differences in Gene Expression.
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Walker DM, Cunningham AM, and Nestler EJ
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- Female, Gene Expression, Humans, Male, Sex Characteristics
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- 2022
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20. Sex-Specific Transcriptional Changes in Response to Adolescent Social Stress in the Brain's Reward Circuitry.
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Walker DM, Zhou X, Cunningham AM, Lipschultz AP, Ramakrishnan A, Cates HM, Bagot RC, Shen L, Zhang B, and Nestler EJ
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- Animals, Brain, Female, Male, Mice, Nucleus Accumbens, Transcriptome, Cocaine, Reward
- Abstract
Background: Sex differences in addiction have been described in humans and animal models. A key factor that influences addiction in both males and females is adolescent experience. Adolescence is associated with higher vulnerability to substance use disorders, and male rodents subjected to adolescent social isolation (SI) stress form stronger preferences for drugs of abuse in adulthood. However, little is known about how females respond to SI, and few studies have investigated the transcriptional changes induced by SI in the brain's reward circuitry., Methods: We tested the hypothesis that SI alters the transcriptome in a persistent and sex-specific manner in prefrontal cortex, nucleus accumbens, and ventral tegmental area. Mice were isolated or group housed from postnatal day P22 to P42, then group housed until ∼P90. Transcriptome-wide changes were investigated by RNA sequencing after acute or chronic cocaine or saline administration., Results: We found that SI disrupts sex-specific transcriptional responses to cocaine and reduces sex differences in gene expression across all three brain regions. Furthermore, SI induces gene expression profiles in males that more closely resemble group-housed females, suggesting that SI "feminizes" the male transcriptome. Coexpression analysis reveals that such disruption of sex differences in gene expression alters sex-specific gene networks and identifies potential sex-specific key drivers of these transcriptional changes., Conclusions: Together, these data show that SI has region-specific effects on sex-specific transcriptional responses to cocaine and provide a better understanding of reward-associated transcription that differs in males and females., (Copyright © 2021 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)
- Published
- 2022
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21. Gene-Targeted, CREB-Mediated Induction of ΔFosB Controls Distinct Downstream Transcriptional Patterns Within D1 and D2 Medium Spiny Neurons.
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Lardner CK, van der Zee Y, Estill MS, Kronman HG, Salery M, Cunningham AM, Godino A, Parise EM, Kim JH, Neve RL, Shen L, Hamilton PJ, and Nestler EJ
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- Animals, Female, Gene Expression, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Neurons metabolism, Receptors, Dopamine D1 metabolism, Cocaine, Nucleus Accumbens metabolism
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Background: The onset and persistence of addiction phenotypes are, in part, mediated by transcriptional mechanisms in the brain that affect gene expression and, subsequently, neural circuitry. ΔFosB is a transcription factor that accumulates in the nucleus accumbens (NAc)-a brain region responsible for coordinating reward and motivation-after exposure to virtually every known rewarding substance, including cocaine and opioids. ΔFosB has also been shown to directly control gene transcription and behavior downstream of both cocaine and opioid exposure, but with potentially different roles in D1 and D2 medium spiny neurons (MSNs) in NAc., Methods: To clarify MSN subtype-specific roles for ΔFosB and investigate how these coordinate the actions of distinct classes of addictive drugs in NAc, we developed a CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9-based epigenome editing tool to induce endogenous ΔFosB expression in vivo in the absence of drug exposure. After inducing ΔFosB in D1- or D2-MSNs or both, we performed RNA sequencing on bulk male and female NAc tissue (n = 6-8/group)., Results: We found that ΔFosB induction elicits distinct transcriptional profiles in NAc by MSN subtype and by sex, establishing for the first time that ΔFosB mediates different transcriptional effects in males versus females. We also demonstrated that changes in D1-MSNs, but not those in D2-MSNs or both, significantly recapitulate changes in gene expression induced by cocaine self-administration., Conclusions: Together, these findings demonstrate the efficacy of a novel molecular tool for studying cell type-specific transcriptional mechanisms and shed new light on the activity of ΔFosB, a critical transcriptional regulator of drug addiction., (Copyright © 2021 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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22. Sperm Transcriptional State Associated with Paternal Transmission of Stress Phenotypes.
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Cunningham AM, Walker DM, Ramakrishnan A, Doyle MA, Bagot RC, Cates HM, Peña CJ, Issler O, Lardner CK, Browne C, Russo SJ, Shen L, and Nestler EJ
- Abstract
Paternal stress can induce long-lasting changes in germ cells potentially propagating heritable changes across generations. To date, no studies have investigated differences in transmission patterns between stress-resilient and stress-susceptible mice. We tested the hypothesis that transcriptional alterations in sperm during chronic social defeat stress (CSDS) transmit increased susceptibility to stress phenotypes to the next generation. We demonstrate differences in offspring from stressed fathers that depend on paternal category (resilient vs susceptible) and offspring sex. Importantly, artificial insemination (AI) reveals that sperm mediates some of the behavioral phenotypes seen in offspring. Using RNA-sequencing (RNA-seq), we report substantial and distinct changes in the transcriptomic profiles of sperm following CSDS in susceptible versus resilient fathers, with alterations in long noncoding RNAs (lncRNAs) predominating especially in susceptibility. Correlation analysis revealed that these alterations were accompanied by a loss of regulation of protein-coding genes by lncRNAs in sperm of susceptible males. We also identify several co-expression gene modules that are enriched in differentially expressed genes (DEGs) in sperm from either resilient or susceptible fathers. Taken together, these studies advance our understanding of intergenerational epigenetic transmission of behavioral experience. SIGNIFICANCE STATEMENT This manuscript contributes to the complex factors that influence the paternal transmission of stress phenotypes. By leveraging the segregation of males exposed to chronic social defeat stress (CSDS) into either resilient or susceptible categories we were able to identify the phenotypic differences in the paternal transmission of stress phenotypes across generations between the two lineages. Importantly, this work also alludes to the significance of both long noncoding RNAs (lncRNAs) and protein coding genes (PCGs) mediating the paternal transmission of stress. The knowledge gained from these data are of particular interest in understanding the risk for the development of psychiatric disorders such as anxiety and depression., (Copyright © 2021 the authors.)
- Published
- 2021
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23. Long read, isoform aware sequencing of mouse nucleus accumbens after chronic cocaine treatment.
- Author
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Estill M, Ribeiro E, Francoeur NJ, Smith ML, Sebra R, Yeh SY, Cunningham AM, Nestler EJ, and Shen L
- Subjects
- Animals, Cocaine pharmacology, Computational Biology methods, Mice, Molecular Sequence Annotation, Sequence Analysis, DNA, Cocaine adverse effects, Gene Expression Profiling, High-Throughput Nucleotide Sequencing, Nucleus Accumbens drug effects, Nucleus Accumbens metabolism, Transcriptome
- Abstract
To better understand the full-length transcriptome of the nucleus accumbens (NAc)-a key brain reward region-in chronic cocaine treatment, we perform the first single molecule, long-read sequencing analysis using the Iso-seq method to detect 42,114 unique transcripts from mouse NAc polyadenylated RNA. Using GENCODE annotation as a reference, we find that over half of the Iso-seq derived transcripts are annotated, while 46% of them harbor novel splicing events in known genes; around 1% of them correspond to other types of novel transcripts, such as fusion, antisense and intergenic. Approximately 34% of the novel transcripts are matched with a compiled transcriptome assembled from published short-read data from various tissues, with the remaining 69% being unique to NAc. These data provide a more complete picture of the NAc transcriptome than existing annotations and can serve as a comprehensive reference for future transcriptomic analyses of this important brain reward region.
- Published
- 2021
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24. Corticosteroids Should Be Used To Treat Slowly Progressive IgA Nephropathy: PRO.
- Author
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Cunningham AM and Reich HN
- Subjects
- Adrenal Cortex Hormones therapeutic use, Glomerular Mesangium, Humans, Glomerulonephritis, IGA drug therapy
- Abstract
Competing Interests: A.M. Cunningham’s work is supported by the Louise Fast Foundation GN fellowship at the University Health Network. H.N. Reich is the national coordinating investigator for Canada for the NEFIGARD study and has received consulting fees from Calliditas. She is a site investigator for studies in IgAN sponsored by Alnylam and Omeros. She has received consulting fees from Novartis, Omeros, and Retrophen.
- Published
- 2021
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25. Paternal transgenerational epigenetic mechanisms mediating stress phenotypes of offspring.
- Author
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Cunningham AM, Walker DM, and Nestler EJ
- Subjects
- Animals, Anxiety genetics, Anxiety Disorders, Female, Humans, Male, Phenotype, Epigenesis, Genetic, Fathers
- Abstract
Depression and anxiety risk are highly influenced by both genetic and environmental factors. Recently, it has been proposed that epigenetic mechanisms may also contribute to the transmission of both depression- and anxiety-related behaviors across multiple generations. This review highlights long-lasting epigenetic alterations observed in offspring of fathers, including some distinct effects on male and female offspring, in animal models. Available evidence emphasizes how both the developmental time point and the type of paternal stress (social vs. asocial) influence the complex transmission patterns of these phenotypes to future generations. This research is critical in understanding the factors that influence risk for depression and anxiety disorders and has the potential to contribute to the development of innovative treatments that can more precisely target vulnerable populations., (© 2019 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.)
- Published
- 2021
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26. Glargine Insulin
- Author
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Cunningham AM and Freeman AM
- Abstract
Insulin glargine is used to manage and treat hyperglycemia in patients with type 1 and type 2 diabetes mellitus. It is a long-acting insulin and is used as basal insulin. This activity will cover the indications, mechanism of action, adverse effects, contraindications, monitoring, and toxicity of insulin glargine pertinent for health care professionals., (Copyright © 2021, StatPearls Publishing LLC.)
- Published
- 2021
27. Understanding and responding to COVID-19 in Wales: protocol for a privacy-protecting data platform for enhanced epidemiology and evaluation of interventions.
- Author
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Lyons J, Akbari A, Torabi F, Davies GI, North L, Griffiths R, Bailey R, Hollinghurst J, Fry R, Turner SL, Thompson D, Rafferty J, Mizen A, Orton C, Thompson S, Au-Yeung L, Cross L, Gravenor MB, Brophy S, Lucini B, John A, Szakmany T, Davies J, Davies C, Thomas DR, Williams C, Emmerson C, Cottrell S, Connor TR, Taylor C, Pugh RJ, Diggle P, John G, Scourfield S, Hunt J, Cunningham AM, Helliwell K, and Lyons R
- Subjects
- COVID-19, Coronavirus Infections epidemiology, Humans, Pneumonia, Viral epidemiology, Risk Factors, SARS-CoV-2, Wales epidemiology, Betacoronavirus, Coronavirus Infections therapy, Delivery of Health Care standards, Pandemics prevention & control, Pneumonia, Viral therapy
- Abstract
Introduction: The emergence of the novel respiratory SARS-CoV-2 and subsequent COVID-19 pandemic have required rapid assimilation of population-level data to understand and control the spread of infection in the general and vulnerable populations. Rapid analyses are needed to inform policy development and target interventions to at-risk groups to prevent serious health outcomes. We aim to provide an accessible research platform to determine demographic, socioeconomic and clinical risk factors for infection, morbidity and mortality of COVID-19, to measure the impact of COVID-19 on healthcare utilisation and long-term health, and to enable the evaluation of natural experiments of policy interventions., Methods and Analysis: Two privacy-protecting population-level cohorts have been created and derived from multisourced demographic and healthcare data. The C20 cohort consists of 3.2 million people in Wales on the 1 January 2020 with follow-up until 31 May 2020. The complete cohort dataset will be updated monthly with some individual datasets available daily. The C16 cohort consists of 3 million people in Wales on the 1 January 2016 with follow-up to 31 December 2019. C16 is designed as a counterfactual cohort to provide contextual comparative population data on disease, health service utilisation and mortality. Study outcomes will: (a) characterise the epidemiology of COVID-19, (b) assess socioeconomic and demographic influences on infection and outcomes, (c) measure the impact of COVID-19 on short -term and longer-term population outcomes and (d) undertake studies on the transmission and spatial spread of infection., Ethics and Dissemination: The Secure Anonymised Information Linkage-independent Information Governance Review Panel has approved this study. The study findings will be presented to policy groups, public meetings, national and international conferences, and published in peer-reviewed journals., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY. Published by BMJ.)
- Published
- 2020
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28. Optogenetic reactivation of prefrontal social neural ensembles mimics social buffering of fear.
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Gutzeit VA, Ahuna K, Santos TL, Cunningham AM, Sadsad Rooney M, Muñoz Zamora A, Denny CA, and Donaldson ZR
- Subjects
- Animals, Fear, Female, Male, Mice, Neurons, Prefrontal Cortex, Rats, Optogenetics, Social Behavior
- Abstract
Social buffering occurs when the presence of a companion attenuates the physiological and/or behavioral effects of a stressful or fear-provoking event. It represents a way in which social interactions can immediately and potently modulate behavior. As such, social buffering is one mechanism by which strong social support increases resilience to mental illness. Although the behavioral and neuroendocrine impacts of social buffering are well studied in multiple species, including humans, the neuronal underpinnings of this behavioral phenomenon remain largely unexplored. Previous work has shown that the infralimbic prefrontal cortex (IL-PFC) is important for processing social information and, in separate studies, for modulating fear and anxiety. Thus, we hypothesized that socially active cells within the IL-PFC may integrate social information to modulate fear responsivity. To test this hypothesis, we employed social buffering paradigms in male and female mice. Similar to prior studies in rats, we found that the presence of a cagemate reduced freezing in fear- and anxiety-provoking contexts. In accordance with previous work, we demonstrated that interaction with a novel or familiar conspecific induces activity in the IL-PFC as evidenced by increased immediate early gene (IEG) expression. We then utilized an activity-dependent tagging murine line, the ArcCreER
T2 mice, to express channelrhodopsin (ChR2) in neurons active during the social encoding of a new cagemate. We found that optogenetic reactivation of these socially active neuronal ensembles phenocopied the effects of cagemate presence in male and female mice in learned and innate fear contexts without being inherently rewarding or altering locomotion. These data suggest that a social neural ensemble within the IL-PFC may contribute to social buffering of fear. These neurons may represent a novel therapeutic target for fear and anxiety disorders.- Published
- 2020
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29. Simple Laboratory Test Utilization Interventions to Reduce Inappropriate Specialty Coagulation Testing.
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Huang H, Cunningham AM, and Harrington AM
- Subjects
- Blood Coagulation Tests economics, Clinical Laboratory Techniques economics, Factor V genetics, Factor Xa Inhibitors blood, Humans, Mutation, Retrospective Studies, Unnecessary Procedures, Blood Coagulation Tests statistics & numerical data, Clinical Laboratory Techniques statistics & numerical data, Factor V analysis, Factor X analysis
- Abstract
Objectives: The naming convention in coagulation may cause confusion in electronic ordering systems, leading to inappropriate test orders. We implemented test utilization efforts and studied utilization before and after interventions for two specialty coagulation assays., Methods: Two interventions were implemented: test names were changed from factor assay to activity, and residents reviewed all factor V and X requests. A retrospective review of factor V and X activity orders was performed for the period 1 year before and after interventions., Results: After interventions, factor V and X activity orders decreased by approximately 40%. Resulted tests decreased by 53.8% and 47.8%, corresponding to reductions of $2,493.05 and $1,867.80 per year in laboratory charges for factor V and factor X activity, respectively. Abnormal factor V activity results increased from 45% to 59%. Factor V activity orders from outpatient clinics decreased by 21.6%., Conclusions: Simple interventions can reduce inappropriate specialty coagulation test orders and unnecessary costs., (© American Society for Clinical Pathology, 2019. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2020
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30. Erythrosin B: a versatile colorimetric and fluorescent vital dye for bacteria.
- Author
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Franke JD, Braverman AL, Cunningham AM, Eberhard EE, and Perry GA
- Subjects
- Cell Membrane chemistry, Colorimetry, Flow Cytometry, Bacteria chemistry, Bacteriological Techniques methods, Erythrosine chemistry, Fluorescent Dyes chemistry
- Abstract
Rapidly assaying cell viability for diverse bacteria species is not always straightforward. In eukaryotes, cell viability is often determined using colorimetric dyes; however, such dyes have not been identified for bacteria. We screened different dyes and found that erythrosin B (EB), a visibly red dye with fluorescent properties, functions as a vital dye for many Gram-positive and -negative bacteria. EB worked at a similar concentration for all bacteria studied and incubations were as short as 5 min. Given EB's spectral properties, diverse experimental approaches are possible to rapidly visualize and/or quantitate dead bacterial cells in a population. As the first broadly applicable colorimetric viability dye for bacteria, EB provides a cost-effective alternative for researchers in academia and industry.
- Published
- 2020
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31. Ancillary Studies in the Diagnostic Evaluation of Large B-Cell Lymphoma.
- Author
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Cunningham AM and Harrington AM
- Subjects
- Diagnosis, Differential, Humans, Lymphoma, Large B-Cell, Diffuse classification, Lymphoma, Large B-Cell, Diffuse diagnosis
- Abstract
Context.—: Large B-cell lymphoma classification has changed significantly over the decades, evolving from a purely morphologic categorization to one using sophisticated ancillary studies including molecular analysis, immunohistochemistry, and cytogenetics, in addition to morphology and clinical presentation., Objective.—: To discuss and interpret the key ancillary studies required for subclassification in 2019 and review the differential diagnosis of diffuse large B-cell lymphoma, not otherwise specified (DLBCL, NOS)., Data Sources.—: Recent literature on the subcategories of large B-cell lymphoma is reviewed, along with relevant updates from the 2016 World Health Organization Classification of Tumours of Hematopoietic and Lymphoid Tissues , with an emphasis on Epstein-Barr virus-positive lymphoproliferative disorders, high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements, and large B-cell lymphoma with IRF4 rearrangement., Conclusions.—: Cases with DLBCL, NOS histology can be further subclassified on the basis of cell of origin studies, Epstein-Barr virus-encoded small RNAs, MYC and BCL2 and/or BCL6 rearrangement studies, and other relevant cytogenetic and immunohistochemical studies. The diagnosis of DLBCL, NOS is therefore a diagnosis of exclusion.
- Published
- 2019
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32. Versican proteolysis predicts immune effector infiltration and post-transplant survival in myeloma.
- Author
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Dhakal B, Pagenkopf A, Mushtaq MU, Cunningham AM, Flietner E, Morrow Z, Papadas A, Hope C, Leith C, Hematti P, Hari P, Callander NS, and Asimakopoulos F
- Subjects
- Biomarkers, Hematopoietic Stem Cell Transplantation adverse effects, Hematopoietic Stem Cell Transplantation methods, Humans, Immunohistochemistry, Multiple Myeloma mortality, Multiple Myeloma therapy, Prognosis, Proteolysis, Treatment Outcome, Tumor Microenvironment, Lymphocytes, Tumor-Infiltrating immunology, Lymphocytes, Tumor-Infiltrating metabolism, Multiple Myeloma immunology, Multiple Myeloma metabolism, Versicans metabolism
- Published
- 2019
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- View/download PDF
33. Vaccination with Tumor-Ganglioside Glycomimetics Activates a Selective Immunity that Affords Cancer Therapy.
- Author
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Tong W, Maira M, Roychoudhury R, Galan A, Brahimi F, Gilbert M, Cunningham AM, Josephy S, Pirvulescu I, Moffett S, and Saragovi HU
- Subjects
- Animals, Antibodies, Monoclonal, Cancer Vaccines therapeutic use, Cell Line, Tumor, Female, Gangliosides therapeutic use, Humans, Immunity, Cellular immunology, Immunity, Humoral immunology, Lymphocytes, Tumor-Infiltrating immunology, Male, Mice, Mice, Inbred C57BL, Neoplasms immunology, Neoplasms therapy, T-Lymphocytes immunology, Vaccination methods, Cancer Vaccines immunology, Gangliosides immunology, Glycolipids immunology
- Abstract
Immune targeting of (glyco)protein tumor markers has been useful to develop cancer and virus vaccines. However, the ganglioside family of tumor-associated glycolipids remains intractable to vaccine approaches. Here we show that synthetic antigens mimicking the carbohydrate moiety of GD2 or GD3 gangliosides can be used as vaccines to activate a selective humoral and cellular immunity that is therapeutic against several cancers expressing GD2 or GD3. Adoptive transfer of T cells generated after vaccination elicits tumor-infiltrating lymphocytes of the γδ T cell receptor and CD8
+ phenotypes; and affords a high therapeutic index. The glycomimetic vaccine principles can be expanded to target the family of tumor gangliosides and other carbohydrates expressed primarily in pathological states., (Crown Copyright © 2019. Published by Elsevier Ltd. All rights reserved.)- Published
- 2019
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34. Functional Interrogation of a Depression-Related Serotonergic Single Nucleotide Polymorphism, rs6295, Using a Humanized Mouse Model.
- Author
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Cunningham AM, Santos TL, Gutzeit VA, Hamilton H, Hen R, and Donaldson ZR
- Subjects
- Alleles, Animals, Behavior, Animal physiology, Depression metabolism, Depressive Disorder metabolism, Disease Models, Animal, Mice, Mice, Transgenic, Motor Activity physiology, Receptor, Serotonin, 5-HT1A metabolism, Brain metabolism, Depression genetics, Depressive Disorder genetics, Polymorphism, Single Nucleotide, Receptor, Serotonin, 5-HT1A genetics
- Abstract
The serotonin 1A receptor (5-HT1A) system has been extensively implicated in modulating mood and behavior. Notably, 5-HT1A levels in humans display remarkable variation, and differences in receptor levels have been linked with a variety of psychiatric disorders. Further, reduction of receptor levels by 30-50% in mice suggests that changes in receptor levels that model existing human variation are sufficient to drive behavioral alterations. As a result, genetic mechanisms that modulate human 5-HT1A levels may be important for explaining individual differences in mood and behavior, representing a potential source of psychiatric disease risk. One common genetic variant implicated in differential 5-HT1A levels is the G/C single nucleotide polymorphism (SNP) rs6295, located upstream of the human 5-HT1A gene. This SNP differentially binds the transcription factor, NUDR/Deaf1, leading to cell-type specific effects on transcription in vitro. To investigate the direct effects of this SNP in the heterogeneous cellular context of the brain, we generated humanized transgenic mice using a design that maximized the local transcriptional landscape of the human HTR1A gene while also controlling for effects of genomic insertion location. We integrated a 180 kb human bacteria artificial chromosome (BAC) transgene containing G- and C-alleles of rs6295 flanked by FRT or loxP sites. Subsequent deletion of each allele by Cre- or Flp-recombinase resulted in rs6295G and C alleles in the same genomic location. These alleles were bred onto a 5-HT1A null mouse such that the human BAC was the sole source of 5-HT1A in these mice. We generated three separate lines, two of which had detectable human 5-HT1A levels in the brain, although none displayed expression in the raphe. Of these, one line exhibited rs6295-dependent differences in 5-HT1A levels and differences in behavior, even though the overall levels were considerably lower than native expression levels. The line-dependent effect of rs6295 on protein levels and behavior may depend upon differences in background genetic factors or different insertion sites across each line. This work confirms that relatively subtle differences in 5-HT1A levels can contribute to differences in behavior and highlights the challenges of modeling human noncoding genetic variation in mice.
- Published
- 2019
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35. Selective Capture and Determination of Receptor-Binding Hemagglutinin in Influenza Vaccine Preparations Using a Coupled Receptor-Binding/RP-HPLC Assay.
- Author
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Lorbetskie B, Cunningham AM, Lemieux M, Durno L, Farnsworth A, Wang J, Li C, Li X, Gilbert M, Sauvé S, and Girard M
- Subjects
- Animals, Birds, Chromatography, High Pressure Liquid methods, Chromatography, Reverse-Phase, Hemagglutinin Glycoproteins, Influenza Virus isolation & purification, Humans, Influenza A Virus, H1N1 Subtype immunology, Influenza A Virus, H1N1 Subtype isolation & purification, Influenza A virus isolation & purification, Influenza in Birds immunology, Influenza in Birds prevention & control, Influenza, Human immunology, Influenza, Human prevention & control, Models, Molecular, N-Acetylneuraminic Acid chemistry, Vaccine Potency, Vaccines, Inactivated immunology, Hemagglutinin Glycoproteins, Influenza Virus immunology, Influenza A virus immunology, Influenza Vaccines immunology
- Abstract
Influenza vaccine potency is determined by the quantification of immunologically active hemagglutinin capable of eliciting neutralizing antibodies upon immunization. Currently, the single radial immunodiffusion (SRID) method is the standard in vitro potency assay used for lot release of seasonal inactivated influenza vaccines. Despite the proven usage of SRID, significant limitations such as the time-consuming preparation of reagents and limited dynamic range warrant the need for the development of alternative potency assays. Such alternative approaches need to discriminate and quantify relevant hemagglutinin material, provide strain identity, and be independent of strain-specific and seasonal reagents. Herein, we present a proof of concept method that combines the capture of conformationally well-folded hemagglutinin via a sialic acid binding step with the resolving power of reversed-phase high-performance liquid chromatography for strain identity and determination. Details of the protocol for the selective capture of receptor-binding hemagglutinin, its release from the receptor, and its relative determination are presented. This approach was found to provide flexibility for the reagents to be used and was adaptable to varying strain compositions of influenza vaccines. This proof of concept approach was developed as an antibody-independent methodology.
- Published
- 2019
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36. Long-Term Behavioral Effects of Post-weaning Social Isolation in Males and Females.
- Author
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Walker DM, Cunningham AM, Gregory JK, and Nestler EJ
- Abstract
Adolescence is a developmental period associated with vast neural and behavioral changes which are accompanied by altered sensitivity to stimuli, both stressful and rewarding. Perturbations, especially stressful stimuli, during this period have been shown to alter behavior in adulthood. Social isolation rearing is one such perturbation. This review highlights the long-term behavioral consequences of adolescent social isolation rearing in rodents with a specific focus on anxiety- and addiction-related behaviors. Sex-specific effects are discussed where data are available. We then consider changes in monoaminergic neurotransmission as one possible mechanism for the behavioral effects described. This research on both normative and perturbed adolescent development is crucial to understanding and treating the increased vulnerability to psychiatric disorders seen in humans during this life stage.
- Published
- 2019
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37. Expression of kappa opioid receptors in developing rat brain - Implications for perinatal buprenorphine exposure.
- Author
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Tan KZ, Cunningham AM, Joshi A, Oei JL, and Ward MC
- Subjects
- Animals, Animals, Newborn, Brain metabolism, Cell Proliferation drug effects, Embryonic Development drug effects, Female, Neural Stem Cells drug effects, Neural Stem Cells metabolism, Neurogenesis drug effects, Neurons drug effects, Neurons metabolism, Oligodendroglia drug effects, Oligodendroglia metabolism, Pregnancy, Rats, Wistar, Receptors, Opioid, kappa antagonists & inhibitors, Analgesics, Opioid toxicity, Brain drug effects, Buprenorphine toxicity, Maternal-Fetal Exchange, Narcotic Antagonists toxicity, Receptors, Opioid, kappa metabolism
- Abstract
Buprenorphine, a mu opioid receptor partial agonist and kappa opioid receptor (KOR) antagonist, is an emerging therapeutic agent for maternal opioid dependence in pregnancy and neonatal abstinence syndrome. However, the endogenous opioid system plays a critical role in modulating neurodevelopment and perinatal buprenorphine exposure may detrimentally influence this. To identify aspects of neurodevelopment vulnerable to perinatal buprenorphine exposure, we defined KOR protein expression and its cellular associations in normal rat brain from embryonic day 16 to postnatal day 23 with double-labelling immunohistochemistry. KOR was expressed on neural stem and progenitor cells (NSPCs), choroid plexus epithelium, subpopulations of cortical neurones and oligodendrocytes, and NSPCs and subpopulations of neurones in postnatal hippocampus. These distinct patterns of KOR expression suggest several pathways vulnerable to perinatal buprenorphine exposure, including proliferation, neurogenesis and neurotransmission. We thus suggest the cautious use of buprenorphine in both mothers and infants until its impact on neurodevelopment is better defined., (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Published
- 2018
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38. Using social media to support small group learning.
- Author
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Cole D, Rengasamy E, Batchelor S, Pope C, Riley S, and Cunningham AM
- Subjects
- Cooperative Behavior, Curriculum, Humans, Information Literacy, Problem-Based Learning trends, Software, Wales, Education, Medical, Undergraduate, Group Processes, Problem-Based Learning methods, Social Media statistics & numerical data, Students, Medical
- Abstract
Background: Medical curricula are increasingly using small group learning and less didactic lecture-based teaching. This creates new challenges and opportunities in how students are best supported with information technology. We explored how university-supported and external social media could support collaborative small group working on our new undergraduate medical curriculum., Methods: We made available a curation platform (Scoop.it) and a wiki within our virtual learning environment as part of year 1 Case-Based Learning, and did not discourage the use of other tools such as Facebook. We undertook student surveys to capture perceptions of the tools and information on how they were used, and employed software user metrics to explore the extent to which they were used during the year., Results: Student groups developed a preferred way of working early in the course. Most groups used Facebook to facilitate communication within the group, and to host documents and notes. There were more barriers to using the wiki and curation platform, although some groups did make extensive use of them. Staff engagement was variable, with some tutors reviewing the content posted on the wiki and curation platform in face-to-face sessions, but not outside these times. A small number of staff posted resources and reviewed student posts on the curation platform., Conclusions: Optimum use of these tools depends on sufficient training of both staff and students, and an opportunity to practice using them, with ongoing support. The platforms can all support collaborative learning, and may help develop digital literacy, critical appraisal skills, and awareness of wider health issues in society.
- Published
- 2017
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39. Standardization of transfusion practice in organ donors using the Digital Intern, an electronic decision support algorithm.
- Author
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Connor JP, Cunningham AM, Raife T, Rose WN, and Medow JE
- Subjects
- Humans, Retrospective Studies, Algorithms, Erythrocyte Transfusion
- Abstract
Background: Prospective clinical trials support restrictive thresholds for red blood cell (RBC) transfusion. Nonsurvivable donors are a major source of organs for transplantation. The Digital Intern (DI) is a computer algorithm to standardize donor care that includes a more restrictive transfusion threshold. The impact of standardized and restrictive RBC transfusion in organ donors, as determined by the DI, has not been reported., Study Design and Methods: We conducted a retrospective cohort study to compare the transfusion practice of the DI (n = 100) to a historic group of physician-managed donors (n = 90). Transfusion rates, the number of units transfused, and pretransfusion laboratory values were compared between groups. The variability of these parameters was also compared between groups. Finally, the number of transplanted organs per donor in each group was compared., Results: The mean time as a donor was 25.9 ± 15.2 hours and was not different between the groups. In the DI group 19% were transfused compared to 26% in the control group (p = 0.3). The number of units transfused was less in the DI group (1 unit vs. 2 units per transfusion, p = 0.03) and the pretransfusion hematocrit was lower in the DI group (23% vs. 27%, p = 0.01). The variability in the latter two parameters was significantly lower in the DI group. The number of transplanted organs per donor was similar in both groups (3.24 [DI] vs. 3.03 [control], p = 0.37)., Conclusion: The DI provides a more standardization transfusion practice in organ donors and reduces blood use without compromising transplantable organs., (© 2017 AABB.)
- Published
- 2017
- Full Text
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40. Characterization of the dTDP-Fuc3N and dTDP-Qui3N biosynthetic pathways in Campylobacter jejuni 81116.
- Author
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Li ZZ, Riegert AS, Goneau MF, Cunningham AM, Vinogradov E, Li J, Schoenhofen IC, Thoden JB, Holden HM, and Gilbert M
- Subjects
- Acyltransferases chemistry, Acyltransferases metabolism, Biosynthetic Pathways genetics, Campylobacter jejuni enzymology, Crystallography, X-Ray, Escherichia coli genetics, Galactose chemistry, Galactose metabolism, Glucose chemistry, Glucose metabolism, Glycosyltransferases chemistry, Glycosyltransferases metabolism, Lipopolysaccharides biosynthesis, Lipopolysaccharides genetics, Nucleotidyltransferases chemistry, Nucleotidyltransferases metabolism, Thymine Nucleotides chemistry, Thymine Nucleotides metabolism, Acyltransferases genetics, Campylobacter jejuni genetics, Galactose genetics, Glycosyltransferases genetics, Nucleotidyltransferases genetics
- Abstract
The Gram-negative bacterium Campylobacter jejuni 81116 (Penner serotype HS:6) has a class E lipooligosaccharide (LOS) biosynthesis locus containing 19 genes, which encode for 11 putative glycosyltransferases, 1 lipid A acyltransferase and 7 enzymes thought to be involved in the biosynthesis of dideoxyhexosamine (ddHexN) moieties. Although the LOS outer core structure of C. jejuni 81116 is still unknown, recent mass spectrometry analyses suggest that it contains acetylated forms of two ddHexN residues. For this investigation, five of the genes encoding enzymes reportedly involved in the biosyntheses of these sugar residues were examined, rmlA, rmlB, wlaRA, wlaRB and wlaRG. Specifically, these genes were cloned and expressed in Escherichia coli, and the corresponding enzymes were purified and tested for biochemical activity. Here we present data demonstrating that RmlA functions as a glucose-1-phosphate thymidylyltransferase and that RmlB is a thymidine diphosphate (dTDP)-glucose 4,6-dehydratase. We also show, through nuclear magnetic resonance spectroscopy and mass spectrometry analyses, that WlaRG, when utilized in coupled assays with either WlaRA or WlaRB and dTDP-4-keto-6-deoxyglucose, results in the production of either dTDP-3-amino-3,6-dideoxy-d-galactose (dTDP-Fuc3N) or dTDP-3-amino-3,6-dideoxy-d-glucose (dTDP-Qui3N), respectively. In addition, the X-ray crystallographic structures of the 3,4-ketoisomerases, WlaRA and WlaRB, were determined to 2.14 and 2.0 Å resolutions, respectively. Taken together, the data reported herein demonstrate that C. jejuni 81116 utilizes five enzymes to synthesize dTDP-Fuc3N or dTDP-Qui3N and that WlaRG, an aminotransferase, can function on sugars with differing stereochemistry about their C-4' carbons. Importantly, the data reveal that C. jejuni 81116 has the ability to synthesize two isomeric ddHexN forms., (© Her Majesty the Queen in Right of Canada 2017. Reproduced with the permission of the Minister of National Research Council of Canada.)
- Published
- 2017
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41. Epidemiology of Bednar tumors in the United States.
- Author
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Liszewski W, Blanchette D, Cunningham AM, and Miller DD
- Subjects
- Adolescent, Adult, Age Distribution, Aged, Aged, 80 and over, Child, Child, Preschool, Dendritic Cells chemistry, Dendritic Cells pathology, Dermatofibrosarcoma pathology, Female, Humans, Incidence, Infant, Male, Melanins analysis, Middle Aged, Organ Specificity, Racial Groups, SEER Program, Skin Neoplasms pathology, United States epidemiology, Young Adult, Dermatofibrosarcoma epidemiology, Skin Neoplasms epidemiology
- Published
- 2016
- Full Text
- View/download PDF
42. The Severity of Gliosis in Hippocampal Sclerosis Correlates with Pre-Operative Seizure Burden and Outcome After Temporal Lobectomy.
- Author
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Johnson AM, Sugo E, Barreto D, Hiew CC, Lawson JA, Connolly AM, Somerville E, Hasic E, Bye AM, and Cunningham AM
- Subjects
- Adult, Antigens, CD metabolism, Antigens, Differentiation, Myelomonocytic metabolism, Case-Control Studies, Child, Cohort Studies, Demography, Female, Glial Fibrillary Acidic Protein metabolism, Gliosis complications, Humans, Magnetic Resonance Imaging, Male, Preoperative Care, Sclerosis complications, Seizures complications, Treatment Outcome, Cost of Illness, Gliosis pathology, Hippocampus pathology, Sclerosis pathology, Seizures surgery, Severity of Illness Index, Temporal Lobe surgery
- Abstract
Astrogliosis and microgliosis in hippocampal sclerosis (HS) are widespread and are postulated to contribute to the pro-excitatory neuropathological environment. This study aimed to establish if seizure burden at the time of surgery or post-surgical outcome were correlated with the extent of gliosis in HS. As a secondary aim, we wanted to determine if the degree of gliosis could be predicted by pre-operative neuroimaging.Children and adults who underwent epilepsy surgery for HS between 2002 and 2011 were recruited (n = 43), and age-matched autopsy controls obtained (n = 15). Temporal lobe specimens were examined by DAB immunohistochemistry for astrocytes (glial fibrillary acidic protein (GFAP)) and microglia (CD68). Cell counting for GFAP and CD68 was performed and quantitative densitometry undertaken for GFAP. Seizure variables and outcome (Engel) were determined through medical record and patient review. Seizure frequency in the 6 months prior to surgery was measured to reflect the acute seizure burden. Duration of seizures, age at onset and age at operation were regarded to reflect chronic seizure burden. Focal, lobar and generalized atrophy on pre-operative MRI were independently correlated with the degree of cortical gliosis in the surgical specimen.In HS, both acute and chronic seizure burden were positively correlated with the degree of gliosis. An increase in reactive astrocyte number in CA3 was the strongest predictor of poor post-operative seizure outcome at 1 and 3 years post-operatively in this cohort. Changes in lower cortical astrocyte and upper cortical microglial number also correlated with post-operative outcome at 1 year. Post-surgical seizure outcome (1, 3 and 5 years) did not otherwise correlate with GFAP immunoreactivity (GFAP-IR) or CD68 immunoreactivity (CD68-IR). Increased microglial activation was detected in patients with pre-operative bilateral convulsive seizures, compared to those without convulsive seizures. Furthermore, focal, lobar and generalized atrophy on pre-operative neuroimaging were independently correlated with the degree of cortical gliosis in the surgical specimen.
- Published
- 2016
- Full Text
- View/download PDF
43. Pathologic Features of Primary Pancreatic Malignancies.
- Author
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Cunningham, AM, Rush PS, and Matkowskyj KA
- Subjects
- Humans, Neuroendocrine Tumors pathology, Pancreatic Neoplasms pathology
- Abstract
This chapter explores the pathologic features of benign and malignant lesions of the pancreas. As pathologic classifications evolve, particularly for cystic lesions and neuroendocrine tumors, it is important for physicians who treat patients with pancreatic neoplasms to fully evaluate these pathologic classifications.
- Published
- 2016
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44. Quantitative magnetic resonance imaging of hepatic steatosis: Validation in ex vivo human livers.
- Author
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Bannas P, Kramer H, Hernando D, Agni R, Cunningham AM, Mandal R, Motosugi U, Sharma SD, Munoz del Rio A, Fernandez L, and Reeder SB
- Subjects
- Adult, Aged, Female, Humans, Magnetic Resonance Spectroscopy, Male, Middle Aged, Non-alcoholic Fatty Liver Disease metabolism, Triglycerides analysis, Liver pathology, Magnetic Resonance Imaging methods, Non-alcoholic Fatty Liver Disease pathology
- Abstract
Unlabelled: Emerging magnetic resonance imaging (MRI) biomarkers of hepatic steatosis have demonstrated tremendous promise for accurate quantification of hepatic triglyceride concentration. These methods quantify the proton density fat-fraction (PDFF), which reflects the concentration of triglycerides in tissue. Previous in vivo studies have compared MRI-PDFF with histologic steatosis grading for assessment of hepatic steatosis. However, the correlation of MRI-PDFF with the underlying hepatic triglyceride content remained unknown. The aim of this ex vivo study was to validate the accuracy of MRI-PDFF as an imaging biomarker of hepatic steatosis. Using ex vivo human livers, we compared MRI-PDFF with magnetic resonance spectroscopy-PDFF (MRS-PDFF), biochemical triglyceride extraction, and histology as three independent reference standards. A secondary aim was to compare the precision of MRI-PDFF relative to biopsy for the quantification of hepatic steatosis. MRI-PDFF was prospectively performed at 1.5 Tesla in 13 explanted human livers. We performed colocalized paired evaluation of liver fat content in all nine Couinaud segments using single-voxel MRS-PDFF (n=117) and tissue wedges for biochemical triglyceride extraction (n=117), and five core biopsies performed in each segment for histologic grading (n=585). Accuracy of MRI-PDFF was assessed through linear regression with MRS-PDFF, triglyceride extraction, and histology. Intraobserver agreement, interobserver agreement, and repeatability of MRI-PDFF and histologic grading were assessed through Bland-Altman analyses. MRI-PDFF showed an excellent correlation with MRS-PDFF (r=0.984, confidence interval 0.978-0.989) and strong correlation with histology (r=0.850, confidence interval 0.791-0.894) and triglyceride extraction (r=0.871, confidence interval 0.818-0.909). Intraobserver agreement, interobserver agreement, and repeatability showed a significantly smaller variance for MRI-PDFF than for histologic steatosis grading (all P<0.001)., Conclusion: MRI-PDFF is an accurate, precise, and reader-independent noninvasive imaging biomarker of liver triglyceride content, capable of steatosis quantification over the entire liver., (© 2015 by the American Association for the Study of Liver Diseases.)
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- 2015
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45. Thou shalt not tweet unprofessionally: an appreciative inquiry into the professional use of social media.
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Pereira I, Cunningham AM, Moreau K, Sherbino J, and Jalali A
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- Canada, Consensus, Education, Humans, Leadership, Physician's Role, Practice Guidelines as Topic, Professional Practice trends, Guideline Adherence, Health Personnel education, Professional Competence standards, Professional Practice standards, Social Media trends, Societies, Medical
- Abstract
Background: Social media may blur the line between socialisation and professional use. Traditional views on medical professionalism focus on limiting motives and behaviours to avoid situations that may compromise care. It is not surprising that social media are perceived as a threat to professionalism., Objective: To develop evidence for the professional use of social media in medicine., Methods: A qualitative framework was used based on an appreciative inquiry approach to gather perceptions and experiences of 31 participants at the 2014 Social Media Summit., Results: The main benefits of social media were the widening of networks, access to expertise from peers and other health professionals, the provision of emotional support and the ability to combat feelings of isolation., Conclusions: Appreciative inquiry is a tool that can develop the positive practices of organisations and individuals. Our results provide evidence for the professional use of social media that may contribute to guidelines to help individuals realise benefits and avoid harms., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.)
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- 2015
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46. Developmental expression of vascular endothelial growth factor receptor 3 and vascular endothelial growth factor C in forebrain.
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Ward MC and Cunningham AM
- Subjects
- Animals, Animals, Newborn, Calcium-Binding Proteins metabolism, Embryo, Mammalian, Female, Male, Microfilament Proteins metabolism, Nerve Tissue Proteins metabolism, Nitric Oxide Synthase Type I metabolism, Phosphopyruvate Hydratase metabolism, Rats, Rats, Wistar, Prosencephalon embryology, Prosencephalon growth & development, Prosencephalon metabolism, Vascular Endothelial Growth Factor C metabolism, Vascular Endothelial Growth Factor Receptor-3 metabolism
- Abstract
Increased understanding of the neurovascular niche suggests that development of the central nervous system (CNS) and its vasculature is coordinated through shared regulatory factors. These include the vascular endothelial growth factor (VEGF) family, reported to promote neuroproliferation and neuroprotection in addition to angiogenesis via its receptors VEGFR1-3. VEGFR3, a mediator of lymphangiogenesis, is expressed in murine and rat brain from early gestation, has been associated with neural progenitors and neurons (Choi et al., 2010) and oligodendroglia (Le Bras et al., 2006) in the developing cortex and is reported to mediate adult neurogenesis in the subventricular zone (SVZ) (Calvo et al., 2011). The early expression pattern of VEGFR3 protein and its cellular associations has not as yet been comprehensively reported. We describe the temporal expression of VEGFR3 protein at a cellular level and its close association with its VEGFC ligand, determined by double-labeling immunohistochemistry in the developing rat brain from embryonic day (E) 13 to postnatal day (P) 23. We found high expression of VEGFR3 in the ventricular zone and along radial glia in early gestation in association with neural stem cells and neuroblasts. Similar expression patterns were seen in the immature olfactory bulb and optic cup. In later development we found less expression by neural progenitors in proliferative regions including the SVZ and dentate gyrus of the hippocampus. In contrast, VEGFR3 expression increased with development in the cortex in neurons and astrocytes, and appeared in the emerging population of oligodendroglial progenitors. High expression in ventricular ependyma, choroid plexus and pigmented retinal epithelium was noted from E18. VEGFC ligand was found in association with VEGFR3 throughout development, with highest expression in embryonic stages. Our findings suggest an important role for VEGFC/VEGFR3 signaling in neuronal proliferation in early forebrain development, and ongoing functions with niche neurogenesis, glial and ependymal function in the maturing postnatal brain., (Copyright © 2015 IBRO. Published by Elsevier Ltd. All rights reserved.)
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- 2015
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47. Cold-Active, Heterotrophic Bacteria from the Highly Oligotrophic Waters of Lake Vanda, Antarctica.
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Vander Schaaf NA, Cunningham AM, Cluff BP, Kraemer CK, Reeves CL, Riester CJ, Slater LK, Madigan MT, and Sattley WM
- Abstract
The permanently ice-covered lakes of the McMurdo Dry Valleys, Antarctica are distinctive ecosystems that consist strictly of microbial communities. In this study, water samples were collected from Lake Vanda, a stratified Dry Valley lake whose upper waters (from just below the ice cover to nearly 60 m) are highly oligotrophic, and used to establish enrichment cultures. Six strains of psychrotolerant, heterotrophic bacteria were isolated from lake water samples from a depth of 50 or 55 m. Phylogenetic analyses showed the Lake Vanda strains to be species of Nocardiaceae, Caulobacteraceae, Sphingomonadaceae, and Bradyrhizobiaceae. All Lake Vanda strains grew at temperatures near or below 0 °C, but optimal growth occurred from 18 to 24 °C. Some strains showed significant halotolerance, but no strains required NaCl for growth. The isolates described herein include cold-active species not previously reported from Dry Valley lakes, and their physiological and phylogenetic characterization broadens our understanding of these limnologically unique lakes.
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- 2015
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48. Response to "About focal cortical dysplasia (FCD) type IIIa".
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Johnson AM, Sugo E, Barreto D, Cunningham AM, Hiew CC, Lawson JA, Somerville ER, Connolly AM, and Bye AM
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- Female, Humans, Male, Epilepsy, Temporal Lobe classification, Epilepsy, Temporal Lobe diagnosis, Malformations of Cortical Development, Group III classification, Malformations of Cortical Development, Group III diagnosis
- Published
- 2014
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49. The intrinsic cysteine and histidine residues of the anti-Salmonella antibody Se155-4: a model for the introduction of new functions into antibody-binding sites.
- Author
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Young NM, Watson DC, Cunningham AM, and MacKenzie CR
- Subjects
- Antibodies, Bacterial genetics, Antibodies, Bacterial immunology, Copper chemistry, Copper metabolism, Cysteine, Enzyme-Linked Immunosorbent Assay, Escherichia coli, Fluorescent Dyes, Histidine, Hydrogen-Ion Concentration, Models, Molecular, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins immunology, Recombinant Proteins metabolism, Antibodies, Bacterial chemistry, Antibodies, Bacterial metabolism, Binding Sites, Antibody, Protein Engineering methods, Salmonella immunology
- Abstract
New functions can be incorporated into anti-hapten or anti-protein antibodies by mutating selected residues in the binding-site region either to Cys, to allow alkylation with reagents bearing the desired functional groups, or to His, to create metal-binding sites or to make antigen binding pH-sensitive. However, choosing suitable sites for these mutations has been hampered by the lack of antibodies with these features, to serve as models. Remarkably, the anti-carbohydrate antibody Se155-4, specific for the Salmonella group B lipopolysaccharide, already has a Cys and two pairs of His residues close to the antigen-binding pocket in its structure, and shows pH-dependent antigen binding. We therefore investigated modification of its Cys94L in an scFv version of the antibody with the aims of creating a 'reagentless' fluorescent sensor and attaching a metal-binding group that might confer lyase activity. These groups were successfully introduced, as judged by mass spectrometry, and had only slightly reduced antigen binding in enzyme-linked immunosorbent assay. The fluorescent product was sensitive to addition of antigen in a solution format, unlike a modification of a more distant Cys introduced into the VH CDR4 loop. Two other routes to modulate antigen binding were also explored, metal binding by the His pair alongside the antigen-binding pocket and insertions into CDR4 to extend the antigen-contact area. His residues adjacent to the antigen-binding pocket bound copper, causing a 5-fold decrease in antigen binding. In CDR4 of the VH domain, the preferred insert length was four residues, which gave stable antigen-binding products but did not improve overall antigen affinity., (© Crown copyright 2014.)
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- 2014
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50. Clinicopathological associations in temporal lobe epilepsy patients utilising the current ILAE focal cortical dysplasia classification.
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Johnson AM, Sugo E, Barreto D, Cunningham AM, Hiew CC, Lawson JA, Somerville ER, Connolly AM, and Bye AM
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- Adult, Child, Cohort Studies, Epilepsy, Temporal Lobe surgery, Female, Humans, Male, Malformations of Cortical Development classification, Malformations of Cortical Development diagnosis, Malformations of Cortical Development surgery, Malformations of Cortical Development, Group III surgery, Retrospective Studies, Treatment Outcome, Epilepsy, Temporal Lobe classification, Epilepsy, Temporal Lobe diagnosis, Malformations of Cortical Development, Group III classification, Malformations of Cortical Development, Group III diagnosis
- Abstract
Objectives: This study utilised the revised 2011 ILAE classification of focal cortical dysplasia (FCD) (Blümcke et al., 2011) to examine pathology in a cohort of children and adults who underwent temporal lobe epilepsy (TLE) surgery, and to describe the electroclinical and imaging features associated with these pathologies., Methods: The sample population were children (n=26) and adults (n=47) who underwent TLE surgery between 2002 and 2011 at our institutions. Neuropathology and MRI studies were re-reviewed by experts blinded to the original diagnosis. EEG and clinical data including current seizure outcome were determined by patient file review and/or patient contact. Pre-operative data, post-operative outcome and pathological diagnoses were compared., Results: The commonest pathology in the adult cohort was isolated hippocampal sclerosis (HS) (n=24, 51.1%) and in the paediatric cohort, isolated tumour (n=10, 38.5%). Overall, HS with associated FCD (FCD IIIA) was the third most common pathology (n=12, 16.4%). Temporal grey matter signal changes on MRI were associated with FCD IIIA (p=0.035). FCD IIIA had the poorest post-surgical seizure outcome compared to all other pathologies (p=0.026). A history of bilateral convulsive seizures was more common in adults (n=40, p<0.0005), and was associated with failure to achieve postoperative seizure freedom (p=0.045). Postoperatively, paediatric TLE had higher rates of seizure freedom (p=0.005) and more children had ceased medication (p<0.0005)., Significance: FCD IIIA is a comparatively common pathological subtype in TLE, with a poor post-surgical outcome. Pre-operative recognition of FCD IIIA may be feasible through grey matter signal change on MRI. Paediatric patients had a higher rate of seizure freedom than adults. Pre-operative bilateral convulsive seizures were associated with poor outcome after surgery., (Crown Copyright © 2014. Published by Elsevier B.V. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
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