Publications by authors named "M Zameel Cader"

92 Publications

Systematic rifampicin resistance errors with Xpert MTB/RIF Ultra: implications for regulation of genotypic assays.

Int J Tuberc Lung Dis 2020 Dec;24(12):1307-1311

Centre for Tuberculosis, National Institute for Communicable Diseases, National Health Laboratory Service, Johannesburg, South Africa, Department of Medical Microbiology, University of Pretoria, Pretoria, South Africa.

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http://dx.doi.org/10.5588/ijtld.20.0396DOI Listing
December 2020

The genetics of migraine and the path to precision medicine.

Authors:
M Zameel Cader

Prog Brain Res 2020 25;255:403-418. Epub 2020 Aug 25.

Translational Molecular Neuroscience Group, Weatherall Institute of Molecular Medicine, Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, United Kingdom. Electronic address:

Migraine is a highly heritable complex brain disorder, imposing a huge burden of disability on sufferers. The genetic architecture of migraine ranges from the rare Mendelian forms whereby a single gene mutation is sufficient to cause disease to gene variants that individually impart only a small increase in migraine risk. Despite the considerable advances in the last decade, there are significant challenges to translate genetic findings into drug targets and eventually successful treatments. The need for such treatments remains, even with the new wave of biological therapies targeting CGRP or the CGRP receptor. This will require integration of genetic data with new technologies such as human stem cell models of migraine that allow the interpretation of genetic risk into disease relevant cellular phenotypes. This was recently undertaken for the first time in migraine, whereby stem cells from patients with the rare TRESK frameshift mutation converted into pain sensory neurons demonstrated hyper-excitability. The continued study of the molecular basis of migraine thus offers new paths to drug targets and precision medicine approaches.
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http://dx.doi.org/10.1016/bs.pbr.2020.06.008DOI Listing
August 2020

TRESK is a key regulator of nocturnal suprachiasmatic nucleus dynamics and light adaptive responses.

Nat Commun 2020 09 14;11(1):4614. Epub 2020 Sep 14.

Translational Molecular Neuroscience Group, Weatherall Institute of Molecular Medicine, Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, UK.

The suprachiasmatic nucleus (SCN) is a complex structure dependent upon multiple mechanisms to ensure rhythmic electrical activity that varies between day and night, to determine circadian adaptation and behaviours. SCN neurons are exposed to glutamate from multiple sources including from the retino-hypothalamic tract and from astrocytes. However, the mechanism preventing inappropriate post-synaptic glutamatergic effects is unexplored and unknown. Unexpectedly we discovered that TRESK, a calcium regulated two-pore potassium channel, plays a crucial role in this system. We propose that glutamate activates TRESK through NMDA and AMPA mediated calcium influx and calcineurin activation to then oppose further membrane depolarisation and rising intracellular calcium. Hence, in the absence of TRESK, glutamatergic activity is unregulated leading to membrane depolarisation, increased nocturnal SCN firing, inverted basal calcium levels and impaired sensitivity in light induced phase delays. Our data reveals TRESK plays an essential part in SCN regulatory mechanisms and light induced adaptive behaviours.
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http://dx.doi.org/10.1038/s41467-020-17978-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7490422PMC
September 2020

Acute Upper GI Bleeding: Good Night, Sleep Tight, Endoscopy Can Wait until Morning Light.

Gastroenterology 2020 Nov 9;159(5):1990-1991. Epub 2020 Sep 9.

MRC Cancer Unit, University of Cambridge, Cambridge, UK.

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http://dx.doi.org/10.1053/j.gastro.2020.09.006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7480222PMC
November 2020

Targeted RNA sequencing enhances gene expression profiling of ultra-low input samples.

RNA Biol 2020 12 28;17(12):1741-1753. Epub 2020 Jun 28.

Department of Psychiatry, University of Oxford , Oxford, UK.

RNA-seq is the standard method for profiling gene expression in many biological systems. Due to the wide dynamic range and complex nature of the transcriptome, RNA-seq provides an incomplete characterization, especially of lowly expressed genes and transcripts. Targeted RNA sequencing (RNA CaptureSeq) focuses sequencing on genes of interest, providing exquisite sensitivity for transcript detection and quantification. However, uses of CaptureSeq have focused on bulk samples and its performance on very small populations of cells is unknown. Here we show CaptureSeq greatly enhances transcriptomic profiling of target genes in ultra-low-input samples and provides equivalent performance to that on bulk samples. We validate the performance of CaptureSeq using multiple probe sets on samples of iPSC-derived cortical neurons. We demonstrate up to 275-fold enrichment for target genes, the detection of 10% additional genes and a greater than 5-fold increase in identified gene isoforms. Analysis of spike-in controls demonstrated CaptureSeq improved both detection sensitivity and expression quantification. Comparison to the CORTECON database of cerebral cortex development revealed CaptureSeq enhanced the identification of sample differentiation stage. CaptureSeq provides sensitive, reliable and quantitative expression measurements on hundreds-to-thousands of target genes from ultra-low-input samples and has the potential to greatly enhance transcriptomic profiling when samples are limiting.
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http://dx.doi.org/10.1080/15476286.2020.1777768DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7746246PMC
December 2020

Patient fibroblast circadian rhythms predict lithium sensitivity in bipolar disorder.

Mol Psychiatry 2020 May 13. Epub 2020 May 13.

Department of Pharmacology, University of Oxford, Mansfield Road, Oxford, OX1 3QT, UK.

Bipolar disorder is a chronic neuropsychiatric condition associated with mood instability, where patients present significant sleep and circadian rhythm abnormalities. Currently, the pathophysiology of bipolar disorder remains elusive, but treatment with lithium continues as the benchmark pharmacotherapy, functioning as a potent mood stabilizer in most, but not all patients. Lithium is well documented to induce period lengthening and amplitude enhancement of the circadian clock. Based on this, we sought to investigate whether lithium differentially impacts circadian rhythms in bipolar patient cell lines and crucially if lithium's effect on the clock is fundamental to its mood-stabilizing effects. We analyzed the circadian rhythms of bipolar patient-derived fibroblasts (n = 39) and their responses to lithium and three further chronomodulators. Here we show, relative to controls (n = 23), patients exhibited a wider distribution of circadian period (p < 0.05), and that patients with longer periods were medicated with a wider range of drugs, suggesting lower effectiveness of lithium. In agreement, patient fibroblasts with longer periods displayed muted circadian responses to lithium as well as to other chronomodulators that phenocopy lithium. These results show that lithium differentially impacts the circadian system in a patient-specific manner and its effect is dependent on the patient's circadian phenotype. We also found that lithium-induced behavioral changes in mice were phenocopied by modulation of the circadian system with drugs that target the clock, and that a dysfunctional clock ablates this response. Thus, chronomodulatory compounds offer a promising route to a novel treatment paradigm. These findings, upon larger-scale validation, could facilitate the implementation of a personalized approach for mood stabilization.
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http://dx.doi.org/10.1038/s41380-020-0769-6DOI Listing
May 2020

In Ovo Delivered Toll-Like Receptor 7 Ligand, Resiquimod Enhances Host Responses against Infectious Bronchitis Corona Virus (IBV) Infection.

Vaccines (Basel) 2020 Apr 15;8(2). Epub 2020 Apr 15.

Department of Ecosystem and Public Health, Faculty of Veterinary Medicine, University of Calgary, Health Research Innovation Center 2C53, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada.

Toll-like receptor (TLR) 7 ligand, resiquimod, has been studied as an adjuvant and antiviral agent against several pathogens in chicken. Yet, the effectiveness of resiquimod against infectious bronchitis virus (IBV) infection has not been evaluated. In this study, we investigated the effectiveness of resiquimod delivered pre-hatch (in ovo) against IBV infection post-hatch identifying key mechanisms involved in resiquimod driven immune activation. First, we found an upregulation of interleukin (IL)-1β and interferon (IFN)-γ mRNA levels and considerable expansions of macrophage and cluster of differentiation (CD) 8α+ T cell populations in lungs of chicken as early as day one post-hatch, following pre-hatch delivery of resiquimod. Second, we observed that resiquimod was able to act as an adjuvant when resiquimod was delivered pre-hatch along with an inactivated IBV vaccine. Finally, when the resiquimod pretreated one-day-old chickens were infected with IBV, reduction in viral shedding via oral and fecal routes was observed at 3 days post- infection. Overall, this study shows that the pre-hatch delivered resiquimod increases cell-mediated immune responses in lungs with an advantage of reduction in IBV shedding.
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http://dx.doi.org/10.3390/vaccines8020186DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7349678PMC
April 2020

Common transcriptional signatures of neuropathic pain.

Pain 2020 07;161(7):1542-1554

Nuffield Department of Clinical Neurosciences, Translational Molecular Neuroscience Group, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom.

Abstarct: The dorsal root ganglia (DRG) are key structures in nociception and chronic pain disorders. Several gene expression studies of DRG in preclinical pain models have been performed, but it is unclear if consistent gene changes are identifiable. We, therefore, compared several recent RNA-Seq data sets on the whole DRG in rodent models of nerve injury. Contrary to previous findings, we show hundreds of common differentially expressed genes and high positive correlation between studies, despite model and species differences. We also find, in contrast to previous studies, that 60% of the common rodent gene response after injury is likely to occur in nociceptors of the DRG. Substantial expression changes are observed at a 1-week time-point, with smaller changes in the same genes at a later 3- to 4-week time-point. However, a subset of genes shows a similar magnitude of changes at both early and late time-points, suggesting their potential involvement in the maintenance of chronic pain. These genes are centred around suppression of endogenous opioid signalling. Reversal of this suppression could allow endogenous and exogenous opioids to exert their analgesic functions and may be an important strategy for treating chronic pain disorders. Currently used drugs, such as amitriptyline and duloxetine, do not seem to appropriately modulate many of the critical pain genes and indeed may transcriptionally suppress endogenous opioid signalling further.
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http://dx.doi.org/10.1097/j.pain.0000000000001847DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7302324PMC
July 2020

Culprit or Bystander: Defective Mitophagy in Alzheimer's Disease.

Front Cell Dev Biol 2019 17;7:391. Epub 2020 Jan 17.

Department of Clinical Molecular Biology, University of Oslo, Akershus University Hospital, Lørenskog, Norway.

Mitophagy is a selective engulfment and degradation of damaged mitochondria through the cellular autophagy machinery, a major mechanism responsible for mitochondrial quality control. Increased accumulation of damaged mitochondria in the Alzheimer's disease (AD) human brain are evident, although underlying mechanisms largely elusive. Recent studies indicate impaired mitophagy may contribute to the accumulation of damaged mitochondria in cross-species AD animal models and in AD patient iPSC-derived neurons. Studies from AD highlight feed-forward vicious cycles between defective mitophagy, and the principal AD pathological hallmarks, including amyloid-β plaques, tau tangles, and inflammation. The concomitant and intertwined connections among those hallmarks of AD and the absence of a real humanized AD rodent model present a challenge on how to determine if defective mitophagy is an early event preceding and causal of Tau/Aβ proteinopathies. Whilst further studies are required to understand these relationships, targeting defective mitophagy holds promise as a new therapeutic strategy for AD.
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http://dx.doi.org/10.3389/fcell.2019.00391DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6978796PMC
January 2020

FAMIN Is a Multifunctional Purine Enzyme Enabling the Purine Nucleotide Cycle.

Cell 2020 01;180(2):278-295.e23

Cambridge Institute of Therapeutic Immunology and Infectious Disease, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge CB2 0AW, UK; Division of Gastroenterology and Hepatology, Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge CB2 0QQ, UK. Electronic address:

Mutations in FAMIN cause arthritis and inflammatory bowel disease in early childhood, and a common genetic variant increases the risk for Crohn's disease and leprosy. We developed an unbiased liquid chromatography-mass spectrometry screen for enzymatic activity of this orphan protein. We report that FAMIN phosphorolytically cleaves adenosine into adenine and ribose-1-phosphate. Such activity was considered absent from eukaryotic metabolism. FAMIN and its prokaryotic orthologs additionally have adenosine deaminase, purine nucleoside phosphorylase, and S-methyl-5'-thioadenosine phosphorylase activity, hence, combine activities of the namesake enzymes of central purine metabolism. FAMIN enables in macrophages a purine nucleotide cycle (PNC) between adenosine and inosine monophosphate and adenylosuccinate, which consumes aspartate and releases fumarate in a manner involving fatty acid oxidation and ATP-citrate lyase activity. This macrophage PNC synchronizes mitochondrial activity with glycolysis by balancing electron transfer to mitochondria, thereby supporting glycolytic activity and promoting oxidative phosphorylation and mitochondrial H and phosphate recycling.
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http://dx.doi.org/10.1016/j.cell.2019.12.017DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6978800PMC
January 2020

High glucose concentrations mask cellular phenotypes in a stem cell model of tuberous sclerosis complex.

Epilepsy Behav 2019 12 21;101(Pt B):106581. Epub 2019 Nov 21.

Oxford Epilepsy Research Group, NIHR Oxford Biomedical Research Centre, Nuffield Department of Clinical Neuroscience, Level 6, West Wing, John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom of Great Britain and Northern Ireland; MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, United Kingdom of Great Britain and Northern Ireland. Electronic address:

Tuberous sclerosis complex (TSC) is a neurodevelopmental disorder caused by deletions in the TSC1 or TSC2 genes that is associated with epilepsy in up to 90% of patients. Seizures are suggested to start in benign brain tumors, cortical tubers, or in the perituberal tissue making these tubers an interesting target for further research into mechanisms underlying epileptogenesis in TSC. Animal models of TSC insufficiently capture the neurodevelopmental biology of cortical tubers, and hence, human stem cell-based in vitro models of TSC are being increasingly explored in attempts to recapitulate tuber development and epileptogenesis in TSC. However, in vitro culture conditions for stem cell-derived neurons do not necessarily mimic physiological conditions. For example, very high glucose concentrations of up to 25 mM are common in culture media formulations. As TSC is potentially caused by a disruption of the mechanistic target of rapamycin (mTOR) pathway, a main integrator of metabolic information and intracellular signaling, we aimed to examine the impact of different glucose concentrations in the culture media on cellular phenotypes implicated in tuber characteristics. Here, we present preliminary data from a pilot study exploring cortical neuronal differentiation on human embryonic stem cells (hES) harboring a TSC2 knockout mutation (TSC2-/-) and an isogenic control line (TSC2+/+). We show that the commonly used high glucose media profoundly mask cellular phenotypes in TSC2-/- cultures during neuronal differentiation. These phenotypes only become apparent when differentiating TSC2+/+ and TSC2-/- cultures in more physiologically relevant conditions of 5 mM glucose suggesting that the careful consideration of culture conditions is vital to ensuring biological relevance and translatability of stem cell models for neurological disorders such as TSC. This article is part of the Special Issue "Proceedings of the 7th London-Innsbruck Colloquium on Status Epilepticus and Acute Seizures".
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http://dx.doi.org/10.1016/j.yebeh.2019.106581DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6943812PMC
December 2019

A causal role for TRESK loss of function in migraine mechanisms.

Brain 2019 12;142(12):3852-3867

Translational Molecular Neuroscience Group, Weatherall Institute of Molecular Medicine, Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, UK.

The two-pore potassium channel, TRESK has been implicated in nociception and pain disorders. We have for the first time investigated TRESK function in human nociceptive neurons using induced pluripotent stem cell-based models. Nociceptors from migraine patients with the F139WfsX2 mutation show loss of functional TRESK at the membrane, with a corresponding significant increase in neuronal excitability. Furthermore, using CRISPR-Cas9 engineering to correct the F139WfsX2 mutation, we show a reversal of the heightened neuronal excitability, linking the phenotype to the mutation. In contrast we find no change in excitability in induced pluripotent stem cell derived nociceptors with the C110R mutation and preserved TRESK current; thereby confirming that only the frameshift mutation is associated with loss of function and a migraine relevant cellular phenotype. We then demonstrate the importance of TRESK to pain states by showing that the TRESK activator, cloxyquin, can reduce the spontaneous firing of nociceptors in an in vitro human pain model. Using the chronic nitroglycerine rodent migraine model, we demonstrate that mice lacking TRESK develop exaggerated nitroglycerine-induced mechanical and thermal hyperalgesia, and furthermore, show that cloxyquin conversely is able to prevent sensitization. Collectively, our findings provide evidence for a role of TRESK in migraine pathogenesis and its suitability as a therapeutic target.
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http://dx.doi.org/10.1093/brain/awz342DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6906598PMC
December 2019

The Role of TRESK in Discrete Sensory Neuron Populations and Somatosensory Processing.

Front Mol Neurosci 2019 17;12:170. Epub 2019 Jul 17.

Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, United Kingdom.

Two-pore domain K (K) channels generate K leak current, which serves a vital role in controlling and modulating neuronal excitability. This diverse family of K channels exhibit distinct expression and function across neuronal tissues. TWIK-related spinal cord K channel (TRESK) is a K channel with a particularly enriched role in sensory neurons and pain pathways. Here, we explored the role of TRESK across molecularly distinct sensory neuron populations and assessed its contribution to different sensory modalities. We found TRESK mRNA only in select populations of C- and A-δ nociceptors, in addition to low threshold D-hair afferents. Neurons from mice in which TRESK has been ablated demonstrated marked hyperexcitability, which was amplified under inflammatory challenge. Detailed behavioral phenotyping of TRESK knockout mice revealed specific deficits in somatosensory processing of noxious and non-noxious stimuli. These results demonstrate novel roles of TRESK in somatosensory processing and offer important information to those wishing to target the channel for therapeutic means.
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http://dx.doi.org/10.3389/fnmol.2019.00170DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6650782PMC
July 2019

Proteolytic shedding of the prion protein via activation of metallopeptidase ADAM10 reduces cellular binding and toxicity of amyloid-β oligomers.

J Biol Chem 2019 04 14;294(17):7085-7097. Epub 2019 Mar 14.

From the Division of Neuroscience and Experimental Psychology, School of Biological Sciences, Faculty of Biology, Medicine and Health, AV Hill Building, University of Manchester, Manchester Academic Health Science Centre, Oxford Road, Manchester M13 9PT,

The cellular prion protein (PrP) is a key neuronal receptor for β-amyloid oligomers (AβO), mediating their neurotoxicity, which contributes to the neurodegeneration in Alzheimer's disease (AD). Similarly to the amyloid precursor protein (APP), PrP is proteolytically cleaved from the cell surface by a disintegrin and metalloprotease, ADAM10. We hypothesized that ADAM10-modulated PrP shedding would alter the cellular binding and cytotoxicity of AβO. Here, we found that in human neuroblastoma cells, activation of ADAM10 with the muscarinic agonist carbachol promotes PrP shedding and reduces the binding of AβO to the cell surface, which could be blocked with an ADAM10 inhibitor. Conversely, siRNA-mediated ADAM10 knockdown reduced PrP shedding and increased AβO binding, which was blocked by the PrP-specific antibody 6D11. The retinoic acid receptor analog acitretin, which up-regulates ADAM10, also promoted PrP shedding and decreased AβO binding in the neuroblastoma cells and in human induced pluripotent stem cell (iPSC)-derived cortical neurons. Pretreatment with acitretin abolished activation of Fyn kinase and prevented an increase in reactive oxygen species caused by AβO binding to PrP Besides blocking AβO binding and toxicity, acitretin also increased the nonamyloidogenic processing of APP. However, in the iPSC-derived neurons, Aβ and other amyloidogenic processing products did not exhibit a reciprocal decrease upon acitretin treatment. These results indicate that by promoting the shedding of PrP in human neurons, ADAM10 activation prevents the binding and cytotoxicity of AβO, revealing a potential therapeutic benefit of ADAM10 activation in AD.
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http://dx.doi.org/10.1074/jbc.RA118.005364DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6497954PMC
April 2019

Mitophagy inhibits amyloid-β and tau pathology and reverses cognitive deficits in models of Alzheimer's disease.

Nat Neurosci 2019 03 11;22(3):401-412. Epub 2019 Feb 11.

Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, Baltimore, MD, USA.

Accumulation of damaged mitochondria is a hallmark of aging and age-related neurodegeneration, including Alzheimer's disease (AD). The molecular mechanisms of impaired mitochondrial homeostasis in AD are being investigated. Here we provide evidence that mitophagy is impaired in the hippocampus of AD patients, in induced pluripotent stem cell-derived human AD neurons, and in animal AD models. In both amyloid-β (Aβ) and tau Caenorhabditis elegans models of AD, mitophagy stimulation (through NAD supplementation, urolithin A, and actinonin) reverses memory impairment through PINK-1 (PTEN-induced kinase-1)-, PDR-1 (Parkinson's disease-related-1; parkin)-, or DCT-1 (DAF-16/FOXO-controlled germline-tumor affecting-1)-dependent pathways. Mitophagy diminishes insoluble Aβ and Aβ and prevents cognitive impairment in an APP/PS1 mouse model through microglial phagocytosis of extracellular Aβ plaques and suppression of neuroinflammation. Mitophagy enhancement abolishes AD-related tau hyperphosphorylation in human neuronal cells and reverses memory impairment in transgenic tau nematodes and mice. Our findings suggest that impaired removal of defective mitochondria is a pivotal event in AD pathogenesis and that mitophagy represents a potential therapeutic intervention.
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http://dx.doi.org/10.1038/s41593-018-0332-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6693625PMC
March 2019

Single stranded (ss)RNA-mediated antiviral response against infectious laryngotracheitis virus infection.

BMC Microbiol 2019 02 8;19(1):34. Epub 2019 Feb 8.

Faculty of Veterinary Medicine, Health Research Innovation Center 2C53, University of Calgary, 3330 Hospital Drive NW, Calgary, AB, T2N 4N1, Canada.

Background: Single stranded ribonucleic acid (ssRNA) binds to toll-like receptor (TLR)7 leading to recruitment of immune cells and production of pro-inflammatory cytokines, which has been shown in mammals. In chickens, synthetic ssRNA analog, resiquimod, has been shown to elicit antiviral response against infectious bursal disease virus infection. The objective of this study was to determine the innate host responses activated by the pre-hatch in ovo administration of resiquimod against infectious laryngotracheitis virus (ILTV) infection in chickens post-hatch.

Results: First, we observed that in ovo treatment of resiquimod at embryo day (ED) 18 increases macrophage recruitment in respiratory and gastrointestinal tissues of chicken day 1 post-hatch in addition to interleukin (IL)-1β in lungs. Second, we observed that in ovo treatment of resiquimod reduces ILTV cloacal shedding at 7 days post-infection (dpi) when challenged at day 1 post-hatch coinciding with higher macrophage recruitment. In vitro, we found that resiquimod enhances production of nitric oxide (NO) and IL-1β and not type 1 interferon (IFN) activity in avian macrophages. Although, the antiviral response against ILTV is associated with the enhanced innate immune response, it is not dependent on any of the innate immune mediators observed as has been shown in vitro using avian macrophage.

Conclusion: This study provides insights into the mechanisms of antiviral response mediated by resiquimod, particularly against ILTV infection in chicken.
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http://dx.doi.org/10.1186/s12866-019-1398-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6368756PMC
February 2019

Responsiveness of the Short Warwick Edinburgh Mental Well-Being Scale (SWEMWBS): evaluation a clinical sample.

Health Qual Life Outcomes 2018 Dec 22;16(1):239. Epub 2018 Dec 22.

Professor of Public Health, Warwick Medical School University of Warwick, Coventry, UK.

Background: SWEMWBS is a popular measure of mental wellbeing, shown to be valid in clinical populations. Responsiveness to change has not yet been formally assessed.

Methods: Analysis of data from a clinical sample of 172 clients undergoing up to 4 sessions of cognitive hypnotherapy. Cohen's D effect size (ES), Standardised response mean (SRM), probability of change statistic (P^) were used to evaluate whether SWEMWBS detected statistically important changes at the group level. Cohen's D effect size (ES) and Standard error of measurement (SEM) and were used to evaluate whether SWEMWBS detected statistically important changes at the individual level.

Results: Mean (SD) SWEMWBS scores increased from baseline to therapy 4 from 19.28 (3.921) to 23.32 (4.873). At group level, using Cohen's D effect size, improvement ranges from ES = 0.20-1.41 and using SRM, ranged from 0.30-0.88, increasing with number of therapy sessions. (P^) ranged from 0.65-0.8. At individual level, use of Cohens D ES > 0.5 indicated statistically important improvement in 29.9-86.1% cf. 20.1-80.6% using a standard of 2.77 SEM (2.87 points). The lower threshold of 1 SEM (1.03 points) indicated statistically important improvement in 43.0-81.0%.

Conclusion: SWEMWBS is responsive to change at individual and group level. At individual level a change of between 1 and 3 points meets thresholds for statisticially important change, depending on standard used. Anchor based studies are necessary to confirm that such change represents minimally important change from the perspective of study participants.
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http://dx.doi.org/10.1186/s12955-018-1060-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6303870PMC
December 2018

Comparative features of infections of two Massachusetts (Mass) infectious bronchitis virus (IBV) variants isolated from Western Canadian layer flocks.

BMC Vet Res 2018 Dec 10;14(1):391. Epub 2018 Dec 10.

Department of Ecosystem and Public Health, Faculty of Veterinary Medicine, University of Calgary, Health Research Innovation Center 2C53, 3330 Hospital Drive NW, Calgary, AB, T2N 4N1, Canada.

Background: Infectious bronchitis virus (IBV) is one of the leading causes of mortality and morbidity in chickens. There are numerous serotypes and variants, which do not confer cross protection resulting in failure of currently used IBV vaccines. Although variant IBV isolates with major genetic differences have been subjected to comparative studies, it is unknown whether minor genetic differences in IBV variants within a serotype are different in terms of pathogenesis and eliciting host responses. Two Massachusetts (Mass) variant IBV isolates recovered from commercial layer flocks in the Western Canadian provinces of Alberta (AB) and Saskatchewan (SK) were compared genetically and evaluated for their pathogenicity, tissue distribution and ability to recruit and replicate in macrophages.

Results: Although whole genome sequencing of these two Mass IBV isolates showed low similarity with the M41 vaccinal strain, they had an identical nucleotide sequence at open reading frames (ORFs) 3a, 3b, envelop (E), matrix (M), 5a and 5b. The rest of the ORFs of these 2 IBV isolates showed 99.9% nucleotide similarity. However, upon experimental infection, we found that the IBV isolate originating from AB was different to the one that originated in SK due to higher tracheal lesion scores and lower lung viral replication and lower genome loads in cecal tonsils. Nevertheless, both IBV isolates elicited host responses characterized by significant macrophage recruitment to the respiratory tract and there was evidence that both IBV isolates replicated within tracheal and lung macrophages.

Conclusions: Overall, this study shows that Mass variant IBV isolates, although possessing minor genetic variations, can lead to significant differences in pathogenicity in young chickens. Further studies are required to investigate the pathogenicity of these two Mass variant IBV isolates in laying hens.
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http://dx.doi.org/10.1186/s12917-018-1720-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6288874PMC
December 2018

Antiviral response elicited against avian influenza virus infection following activation of toll-like receptor (TLR)7 signaling pathway is attributable to interleukin (IL)-1β production.

BMC Res Notes 2018 Dec 4;11(1):859. Epub 2018 Dec 4.

Faculty of Veterinary Medicine, University of Calgary, Health Research Innovation Center 2C53, 3330 Hospital Drive NW, Calgary, AB, T2N 4N1, Canada.

Objective: Single stranded ribonucleic acid (ssRNA) binds to toll-like receptor (TLR)7 leading to recruitment of immune cells and production of pro-inflammatory cytokines, which has been shown in mammals. In chickens, ssRNA has been shown to elicit antiviral response against infectious bursal disease virus infection. The objectives of this study were to determine the pro-inflammatory mediators that are activated downstream of TLR7 signaling pathway in avian macrophages and their roles in antiviral response against avian influenza virus (AIV) infection.

Results: In this study, first, we stimulated avian macrophages with the analog of ssRNA, resiquimod, and found that the ssRNA was capable of increasing nitric oxide (NO) and interleukin (IL-1β) production in avian macrophages. Second, we observed when the avian macrophages were stimulated with ssRNA, it elicits an antiviral response against AIV. Finally, we demonstrated that when we blocked the IL-1β response using IL-1 receptor antagonist (IL-1Ra) and the NO production using a selective inhibitor of inducible nitric oxide synthase (iNOS), N-([3-(aminomethyl)phenyl]methyl)ethanimidamide dihydrochloride (1400 W), the antiviral response against AIV is attributable to IL-1β production and not to the NO production. This study provides insights into the mechanisms of antiviral response mediated by ssRNA, particularly against AIV infection.
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http://dx.doi.org/10.1186/s13104-018-3975-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6280464PMC
December 2018

The In Ovo Delivery of CpG Oligonucleotides Protects against Infectious Bronchitis with the Recruitment of Immune Cells into the Respiratory Tract of Chickens.

Viruses 2018 11 15;10(11). Epub 2018 Nov 15.

Faculty of Veterinary Medicine, University of Calgary, Health Research Innovation Center, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada.

The in ovo delivery of cytosine-guanosine (CpG) oligodeoxynucleotides (ODNs) protects chickens against many bacterial and viral infections, by activating the toll-like receptor (TLR)21 signaling pathway. Although the delivery of CpG ODNs in ovo at embryo day (ED) 18 has been shown to reduce infectious bronchitis virus (IBV) loads in embryonic chicken lungs pre-hatch, whether in ovo delivered CpG ODNs are capable of protecting chickens against a post-hatch challenge is unknown. Thus, our objectives were to determine the protective effect of the in ovo delivery of CpG ODNs at ED 18 against IBV infection encountered post-hatch and, then, to investigate the mechanisms of protection. We found significantly higher survival rates and reduced IBV infection in the chickens following the pre-treatment of the ED 18 eggs with CpG ODNs. At 3 days post infection (dpi), we found an increased recruitment of macrophages, cluster of differentiation (CD)8α+ and CD4+ T lymphocytes, and an up-regulation of interferon (IFN)-γ mRNA in the respiratory tract of the chickens. Overall, it may be inferred that CpG ODNs, when delivered in ovo, provide protection against IBV infection induced morbidity and mortality with an enhanced immune response.
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http://dx.doi.org/10.3390/v10110635DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6266937PMC
November 2018

The Virtuous Interplay of Infrastructure Development and the Complexity of Nations.

Entropy (Basel) 2018 Oct 3;20(10). Epub 2018 Oct 3.

Country Analytics, International Finance Corporation-World Bank Group, Washington, DC 20433, USA.

Does the infrastructure stock catalyze the development of new capabilities and ultimately of new products or vice-versa? Here we want to quantify the interplay between these two dimensions from a temporal dynamics perspective and, namely, to address whether the interaction occurs predominantly in a specific direction. We therefore need to measure the complexity of an economy (i.e., its capability stock) and the infrastructure stock of a country. For the former, we leverage a previously proposed metrics, the Economic Fitness (Tacchella, A.; et al. , , 723). For the latter, we propose a new purely statistical indicator which is the performed on the 47 infrastructure indicators published by the World Bank. The proposed indicator still belongs to the class of linear combination of relevant indicators but, differently from standard economic indicators of the same type as the , the , etc, the weights of the linear combination are not subjectively chosen or re-calibrated on a regular basis but they are those which capture the highest fraction of the information encoded in the initial dataset. The two metrics allow the study of the dynamics in the plane and reveal the existence of two regimes: one for low Fitness where the infrastructure and the complexity of an economy are unrelated and a second regime where the two dimensions are tightly related. To quantify the interplay of the two dimensions in this latter regime, we assume a parsimonious linear dynamic model and the emerging picture is that: (i) the feedback occurs in both directions; (ii) on the short-term (<3 years) the predominant direction of interaction is from infrastructure to capability stock; (iii) while for longer time scale (>3 years) the interaction is reversed, new capabilities lead to increasing infrastructure stock.
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http://dx.doi.org/10.3390/e20100761DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7512323PMC
October 2018

Country Case Studies in Economic Fitness: Mexico and Brazil.

Entropy (Basel) 2018 Oct 1;20(10). Epub 2018 Oct 1.

International Finance Corporation, World Bank Group, Washington, DC 20433, USA.

We leverage a new complexity framework called Economic Fitness, which characterizes an economy's level of diversification and its capabilities to produce more complex products. It can be used to predict economic growth and competitiveness. This paper describes an application of Economic Fitness called the Country Opportunity Spotlight (COS) that assesses a country's current level of capabilities and demonstrates which industries have upgrade and diversification potential given those capabilities. It helps unlock the explanatory and predictive power of Economic Fitness for policymakers. COS results serve as a starting point for policymakers to shape and validate priorities, compare countries, asses the capabilities needed in specific industries and begin identifying constraints to growth. We showcase the use of this framework for Mexico and Brazil. These countries provide an interesting case study, as they have similar growth outlooks yet demonstrate different productive capabilities. Examining Mexico and Brazil side by side illustrates the value this analysis can have on deciphering structural change and decision making and at the same time reinforces the need for a nuanced consideration of each country's unique context.
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http://dx.doi.org/10.3390/e20100753DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7512315PMC
October 2018

Reproducibility of Molecular Phenotypes after Long-Term Differentiation to Human iPSC-Derived Neurons: A Multi-Site Omics Study.

Stem Cell Reports 2018 10 20;11(4):897-911. Epub 2018 Sep 20.

Neuroscience Discovery, Biology Department, AbbVie Deutschland GmbH & Co. KG, Ludwigshafen, Germany. Electronic address:

Reproducibility in molecular and cellular studies is fundamental to scientific discovery. To establish the reproducibility of a well-defined long-term neuronal differentiation protocol, we repeated the cellular and molecular comparison of the same two iPSC lines across five distinct laboratories. Despite uncovering acceptable variability within individual laboratories, we detect poor cross-site reproducibility of the differential gene expression signature between these two lines. Factor analysis identifies the laboratory as the largest source of variation along with several variation-inflating confounders such as passaging effects and progenitor storage. Single-cell transcriptomics shows substantial cellular heterogeneity underlying inter-laboratory variability and being responsible for biases in differential gene expression inference. Factor analysis-based normalization of the combined dataset can remove the nuisance technical effects, enabling the execution of robust hypothesis-generating studies. Our study shows that multi-center collaborations can expose systematic biases and identify critical factors to be standardized when publishing novel protocols, contributing to increased cross-site reproducibility.
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http://dx.doi.org/10.1016/j.stemcr.2018.08.013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6178242PMC
October 2018

Shell-Less Egg Syndrome (SES) Widespread in Western Canadian Layer Operations Is Linked to a Massachusetts (Mass) Type Infectious Bronchitis Virus (IBV) Isolate.

Viruses 2018 08 18;10(8). Epub 2018 Aug 18.

Department of Ecosystem and Public Health, Faculty of Veterinary Medicine, University of Calgary, Health Research Innovation Center 2C53, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada.

A disease with a sudden drop in egg production and shell-less eggs called, shell-less egg syndrome (SES) has been observed in Western Canada egg layer flocks since 2010. The etiology of this disease is not known. We hypothesize that SES is caused by an infectious bronchitis virus (IBV) strain since it is known that IBV replicates in the shell gland causing various eggshell abnormalities. In this study, we screened egg layer flocks, in the provinces of Alberta (AB) and Saskatchewan (SK), with and without a history of SES for the presence of IBV infection. During 2015⁻2016, a total of 27 egg layer flocks were screened in AB ( = 7) and SK ( = 20). Eighty-one percent of the screened flocks ( = 22) were positive for IBV infection. Thirty of these isolates were successfully characterized using molecular tools targeting the most variable () 1 gene. IBV isolates from this study clustered into three genotypes based on partial gene variability. The majority of the IBV isolates (70%) were Massachusetts (Mass) type, and the rest were either Connecticut (Conn) type or an uncharacterized genotype with genetic characteristics of Mass and Conn types. Since the majority of the IBV isolates included within the Mass type, we used a Mass type IBV isolate to reproduce SES in specific pathogen free (SPF) white leghorn chickens in lay. Further studies are warranted to investigate whether other IBV isolates can cause SES, to clarify the pathogenesis of SES and to develop a vaccine in order to prevent SES as observed in Western Canadian layer flocks.
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http://dx.doi.org/10.3390/v10080437DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6116215PMC
August 2018

Toll-like receptor (TLR)4 signalling induces myeloid differentiation primary response gene (MYD) 88 independent pathway in avian species leading to type I interferon production and antiviral response.

Virus Res 2018 09 8;256:107-116. Epub 2018 Aug 8.

Department of Ecosystem and Public Health, Faculty of Veterinary Medicine, University of Calgary, Health Research Innovation Center 2C53, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, Canada. Electronic address:

Engagement of toll-like receptor (TLR)4 ligand, lipopolysaccharide (LPS) with TLR4 in mammals activates two downstream intracellular signaling routes; the myeloid differentiation primary response gene (MyD)88 dependent and independent pathways. However, existence of the later pathway leading to production of type I interferons (IFNs) in avian species has been debated due to conflicting observations. The objective of our study was to investigate whether LPS induces type I IFN production in chicken macrophages leading to antiviral response attributable to type I IFN. We found that LPS elicits type I IFN response dominated by IFN-β production. We also found that reduction in infectious laryngotracheitis virus (ILTV) replication by LPS-mediated antiviral response is attributable to type I IFNs in addition to nitric oxide (NO). Our findings imply that LPS elicits both MyD88 dependent and independent pathways in chicken macrophages consequently eliciting anti-ILTV response attributable to production of both type I IFNs and NO.
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http://dx.doi.org/10.1016/j.virusres.2018.08.008DOI Listing
September 2018

Double-Stranded Ribonucleic Acid-Mediated Antiviral Response Against Low Pathogenic Avian Influenza Virus Infection.

Viral Immunol 2018 Jul/Aug;31(6):433-446. Epub 2018 May 29.

1 Department of Ecosystem and Public Health, Faculty of Veterinary Medicine, Health Research Innovation Center 2C53, University of Calgary , Calgary, Alberta, Canada .

Toll-like receptor (TLR)3 signaling pathway is known to induce type 1 interferons (IFNs) and proinflammatory mediators leading to antiviral response against many viral infections. Double-stranded ribonucleic acid (dsRNA) has been shown to act as a ligand for TLR3 and, as such, has been a focus as a potential antiviral agent in many host-viral infection models. Yet, its effectiveness and involved mechanisms as a mediator against low pathogenic avian influenza virus (LPAIV) have not been investigated adequately. In this study, we used avian fibroblasts to verify whether dsRNA induces antiviral response against HN LPAIV and clarify whether type 1 IFNs and proinflammatory mediators such as interleukin (IL)-1β are contributing to the dsRNA-mediated antiviral response against HN LPAIV. We found that dsRNA induces antiviral response in avian fibroblasts against HN LPAIV infection. The treatment of avian fibroblasts with dsRNA increases the expressions of TLR3, IFN-α, IFN-β, and IL-1β. We also confirmed that this antiviral response elicited against HN LPAIV infection correlates, but is not attributable to type 1 IFNs or IL-1β. Our findings imply that the TLR3 ligand, dsRNA, can elicit antiviral response in avian fibroblasts against LPAIV infection, highlighting potential value of dsRNA as an antiviral agent against LPAIV infections. However, further investigations are required to determine the potential role of other innate immune mediators or combination of the tested cytokines in the dsRNA-mediated antiviral response against HN LPAIV infection.
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http://dx.doi.org/10.1089/vim.2017.0142DOI Listing
October 2018

Small vessels, dementia and chronic diseases - molecular mechanisms and pathophysiology.

Clin Sci (Lond) 2018 04 30;132(8):851-868. Epub 2018 Apr 30.

Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow, U.K.

Cerebral small vessel disease (SVD) is a major contributor to stroke, cognitive impairment and dementia with limited therapeutic interventions. There is a critical need to provide mechanistic insight and improve translation between pre-clinical research and the clinic. A 2-day workshop was held which brought together experts from several disciplines in cerebrovascular disease, dementia and cardiovascular biology, to highlight current advances in these fields, explore synergies and scope for development. These proceedings provide a summary of key talks at the workshop with a particular focus on animal models of cerebral vascular disease and dementia, mechanisms and approaches to improve translation. The outcomes of discussion groups on related themes to identify the gaps in knowledge and requirements to advance knowledge are summarized.
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http://dx.doi.org/10.1042/CS20171620DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6700732PMC
April 2018

Low pathogenic avian influenza virus infection increases the staining intensity of KUL01+ cells including macrophages yet decrease of the staining intensity of KUL01+ cells using clodronate liposomes did not affect the viral genome loads in chickens.

Vet Immunol Immunopathol 2018 Apr 23;198:37-43. Epub 2018 Feb 23.

Department of Ecosystem and Public Health, Faculty of Veterinary Medicine, University of Calgary, Health Research Innovation Center 2C53, 3330 Hospital Drive NW, Calgary, AB, T2N 4N1, Canada. Electronic address:

The effect of depletion of macrophages using clodronate liposomes as well as macrophage response following viral infections have been studied in various mouse-virus infection models, but they have not been extensively studied in chickens relevant to virus infections. When we infected day 6 chickens with H4N6 low pathogenic avian influenza virus (LPAIV), we observed that H4N6 LPAIV infection increased the staining intensity of KUL01+ cells in trachea, lungs and duodenum of chickens at 3 days post-infection. Then, we used clodronate liposomes intra-abdominally in 5 day-old chickens and found significant reduction of staining intensity of KUL01+ cells in trachea and duodenum but not in lungs at 4 days post-treatment. When we infected the clodronate liposome and PBS liposome treated chickens with H4N6 LPAIV intra-nasally at day 6, we found no effect on H4N6 LPAIV genome loads in trachea, lungs and duodenum of chickens. This study indicates that although KUL01+ cell intensity are increased in respiratory and gastrointestinal tissues in chickens following H4N6 LPAIV infection, the decrease of KUL01+ cell intensity using clodronate liposomes did not change the H4N6 LPAIV genome loads in any of the examined tissues suggesting that KUL01+ cells may not be critical during H4N6 LPAIV infection in chicken.
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http://dx.doi.org/10.1016/j.vetimm.2018.02.009DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7112785PMC
April 2018

Potential mediators of in ovo delivered double stranded (ds) RNA-induced innate response against low pathogenic avian influenza virus infection.

Virol J 2018 03 12;15(1):43. Epub 2018 Mar 12.

Department of Ecosystem and Public Health, University of Calgary, Health Research Innovation Center 2C53, 3330 Hospital Drive NW, Calgary, AB, T2N 4N1, Canada.

Background: Toll like receptor (TLR) 3 is a critically important innate pattern recognizing receptor that senses many viral infections. Although, it has been shown that double stranded (ds) RNA can be used for the stimulation of TLR3 signaling pathway in a number of host-viral infection models, it's effectiveness as an antiviral agent against low pathogenic avian influenza virus (LPAIV) needs further investigation.

Methods: In this study, first, we delivered TLR3 ligand, dsRNA, in ovo at embryo day (ED)18 since in ovo route is routinely used for vaccination against poultry viral and parasitic infections and infected with H4N6 LPAIV 24-h post-treatment. A subset of in ovo dsRNA treated and control groups were observed for the expressions of TLR3 and type I interferon (IFN)s, mRNA expression of interleukin (IL)-1β and macrophage recruitment coinciding with the time of H4N6 LPAIV infection (24 h post-treatment). Additionally, Day 1 chickens were given dsRNA intra-tracheally along with a control group and a subset of chickens were infected with H4N6 LPAIV 24-h post-treatment whereas the rest of the animals were observed for macrophage and type 1 IFN responses coinciding with the time of viral infection.

Results: Our results demonstrate that the pre-hatch treatment of eggs with dsRNA reduces H4N6 replication in lungs. Further studies revealed that in ovo delivery of dsRNA increases TLR3 expression, type I IFN production and number of macrophages in addition to mRNA expression of IL-1β in lung 24-h post-treatment. The same level of induction of innate response was not evident in the spleen. Moreover, we discovered that dsRNA elicits antiviral response against LPAIV correlating with type I IFN activity in macrophages in vitro. Post-hatch, we found no difference in H4N6 LPAIV genome loads between dsRNA treated and control chickens although we observed higher macrophage recruitment and IFN-β response coinciding with the time of viral infection.

Conclusions: Our findings imply that the TLR3 ligand, dsRNA has antiviral activity in ovo and in vitro but not in chickens post-hatch and dsRNA-mediated innate host response is characterized by macrophage recruitment and expressions of TLR3 and type 1 IFNs.
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http://dx.doi.org/10.1186/s12985-018-0954-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5848551PMC
March 2018