Mechanical nociceptive threshold testing in Bos indicus bull calves

The aim of this prospective, controlled, randomised trial was to develop a technique for mechanical nociceptive threshold testing (NTT) to assess pain in Bos indicus bull calves undergoing surgical castration. Analgesia was provided by 0.5 mg/kg subcutaneous (SC) meloxicam (M) and/or 2 mg/kg of intra-testicular and SC (at the surgery sites) lidocaine (L). Forty-eight Brahman bull calves at 6–8 months of age were divided into six study groups, each with eight animals: no surgery control; surgical castration (C) without analgesia; C and Mpre-op; C and Mpost-op; C, L and Mpost-op; C and L. Mechanical NTT was performed the day before surgery (Day –1) and on Days 1, 2, 6, 10 and 13 after surgery. A handheld manual pneumatic device with a 1-mm (diameter) blunt pin was used to deliver a mechanical stimulus to a maximum of 27 Newtons either side of the most dorsal aspect of the sacrum. The most frequent responses to the mechanical stimulus were lifting or kicking of the leg on the same side as the stimulus (31%) and stepping away from the stimulus (24.9%). Data were analysed with a mixed effect linear model with the nociceptive threshold (NT) as the response variable and day and analgesic treatment as predictors (P < 0.05 was considered significant). For all groups, there was a trend towards decreasing NT over the study period but there were no significant differences between groups. Step down model selection with day, batch and treatment terms revealed a significant effect of day (P < 0.001) and batch (P = 0.007). Mechanical NTT for assessment of pain in Bos indicus bull calves requires further refinement to determine if this is a useful method of pain assessment

The one comparing narrative social network extraction techniques

Analysing narratives through their social networks is an expanding field in quantitative literary studies. Manually extracting a social network from any narrative can be time consuming, so automatic extraction methods of varying complexity have been developed. However, the effect of different extraction methods on the resulting networks is unknown. Here we model and compare three extraction methods for social networks in narratives: manual extraction, co-occurrence automated extraction and automated extraction using machine learning. Although the manual extraction method produces more precise results in the network analysis, it is highly time consuming. The automatic extraction methods yield comparable results for density, centrality measures and edge weights. Our results provide evidence that automatically-extracted social networks are reliable for many analyses. We also describe which aspects of analysis are not reliable with such a social network. Our findings provide a framework to analyse narratives, which help us improve our understanding of how stories are written and evolve, and how people interact with each other. Index Tenns-social networks, narratives, televisionMichelle Edwards, Jonathan Tuke, Matthew Roughan, Lewis Mitchel

The splice site variant rs11078928 may be associated with a genotype-dependent alteration in expression of GSDMB transcripts.

Published onlineJournal ArticleResearch Support, Non-U.S. Gov'tBACKGROUND: Many genetic variants have been associated with susceptibility to complex traits by genome wide association studies (GWAS), but for most, causal genes and mechanisms of action have yet to be elucidated. Using bioinformatics, we identified index and proxy variants associated with autoimmune disease susceptibility, with the potential to affect splicing of candidate genes. PCR and sequence analysis of whole blood RNA samples from population controls was then carried out for the 8 most promising variants to determine the effect of genetic variation on splicing of target genes. RESULTS: We identified 31 splice site SNPs with the potential to affect splicing, and prioritised 8 to determine the effect of genotype on candidate gene splicing. We identified that variants rs11078928 and rs2014886 were associated with altered splicing of the GSDMB and TSFM genes respectively. rs11078928, present in the asthma and autoimmune disease susceptibility locus on chromosome 17q12-21, was associated with the production of a novel Δ exon5-8 transcript of the GSDMB gene, and a separate decrease in the percentage of transcripts with inclusion of exon 6, whereas the multiple sclerosis susceptibility variant rs2014886, was associated with an alternative TFSM transcript encompassing a short cryptic exon within intron 2. CONCLUSIONS: Our findings demonstrate the utility of a bioinformatic approach in identification and prioritisation of genetic variants effecting splicing of their host genes, and suggest that rs11078928 and rs2014886 may affect the splicing of the GSDMB and TSFM genes respectively.Mendip Golf ClubNIHR Exeter Clinical Research Facilit

Mitigation of electroencephalographic and cardiovascular responses to castration in Bos indicus bulls following the administration of either lidocaine or meloxicam

Objective To investigate the mitigating effects of administration of local or systemic meloxicam on the electroencephalographic (EEG) and cardiovascular responses during surgical castration of Bos indicus bull calves. Study design Prospective, randomized, experimental study. Animals Thirty-six 6–8 month-old Bos indicus bull calves, with a mean ± standard deviation weight of 237 ± 19 kg. Methods Animals were randomly allocated to three groups of 12 (group L, 260 mg of 2% lidocaine subcutaneously and intratesticularly 5 minutes prior to castration; group M, 0.5 mg kg−1 of meloxicam subcutaneously 30 minutes prior to castration; group C, no pre-operative analgesia administered). Anaesthesia was induced and maintained with halothane (0.9–1.1%) in oxygen. Electroencephalogram, heart rate (HR) and mean blood pressure (MAP) were recorded for 300 seconds prior to (baseline, B) and from the start of surgery (first testicle incision, T1). HR and MAP were compared at 10 second intervals for 90 seconds from the start of T1. Median frequency (F50), spectral edge frequency (F95) and total power of the EEG (Ptot) were analysed using area under the curve comparing T1 to B. Results All EEG variables were significantly different between B and T1 (p ≤ 0.0001). No differences in F50 were found between groups during T1 (p = 0.6491). F95 and Ptot were significantly different between group L and groups C and M during T1 (p = 0.0005 and 0.0163, respectively). There were transient significant changes in HR and MAP in groups L and M compared to group C during the 20–50 second periods. Conclusions The EEG changes indicate nociceptive responses in all three groups during surgical castration, greater in group L compared to groups C and M. Both analgesics attenuated the peracute cardiovascular response. Lidocaine and meloxicam administered prior to castration attenuated these responses in Bos indicus bull calves. Clinical relevance These findings provide support for the pre-operative administration of lidocaine and potentially meloxicam for castration in Bos indicus bull calves

Gender-specific effects of COVID-19 lockdowns on scientific publishing productivity: Impact and resilience

Available online 3 February 2023Rationale: The SARS-CoV2 pandemic led to drastic social restrictions globally. Early data suggest that women in science have been more adversely affected by these lockdowns than men, with relatively fewer scientific articles authored by women. However, these observations test broad populations with many potential causes of disparity. Australia presents a natural experimental condition where several states of similar demographics and disease impact had differing approaches in their social isolation strategies. The state of Victoria experienced 280 days of lockdowns from 2020 to 2021, whereas the comparable state of New South Wales experienced 107 days, most of these in 2021, and other states even fewer restrictions. Objective and methods: To assess how the gender balance changed in Australian biomedical publishing with the lockdowns, we created a custom workflow to analyse PubMed data from more than 120,000 published articles submitted in 2019–2021 from Australian authors. Results: Broadly, Australian women have been incredibly resilient to the challenges faced by the lockdowns. There was an increase in the number of published articles submitted in 2020 that was equally due to women as men, including from Victoria. On the other hand, articles specifically addressing COVID-19 were significantly less likely to be authored by women than those on other topics, a finding not likely due to particular gender imbalance in virology or viral epidemiology, since publications on HIV followed similar patterns to previous years. By 2021, this imbalance had reversed, with more COVID-19-related papers authored by women than men. Conclusions: These data suggest women from Victoria were less able to rapidly transition to new research early in the pandemic but had accommodated to the new conditions by 2021. This work indicates we need strategies to support women in science as the pandemic continues and to continue to monitor the situation for its impact on vulnerable groups.M. Ryan, J. Tuke, M.R. Hutchinson, S.J. Spence

Toxic Stress and Quality of Life in Early School‐Aged Ugandan Children With and Without Perinatal Human Immunodeficiency Virus Infection

Caregiver’s and child’s self‐reported quality of life (QOL) was defined using standardized questionnaires in a sample (N = 277) of 6–10 years old HIV‐infected, HIV‐exposed uninfected, and HIV‐unexposed uninfected children from Uganda. Psychosocial stress (acute stress and cumulative lifetime adversity) and physiologic stress (dysregulations across 13 biomarkers), perinatal HIV status, and their interaction were related to child QOL via general linear models. Lower child‐ and caregiver‐reported psychosocial stress were dose‐dependently associated with higher QOL (acute stress: mean difference coefficient b = 8.1–14.8, effect size [ES] = 0.46–0.83). Lower allostasis was dose‐dependently associated with higher QOL (b = 6.1–9.7, ES = 0.34–0.54). Given low caregiver acute stress, QOL for HIV‐infected was similar to HIV‐uninfected children; however, given high caregiver acute stress, a QOL disadvantage (b = −7.8, 95% CI: −12.8, −2.8; ES = −0.73) was evident for HIV‐infected versus uninfected children. Testing of caregiver stress reduction interventions is warranted to increase wellbeing in dependent children.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/156459/2/cad20355.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/156459/1/cad20355_am.pd

Sex differences in mechanical allodynia: how can it be preclinically quantified and analyzed?

Extent: 16 p.Translating promising preclinical drug discoveries to successful clinical trials remains a significant hurdle in pain research. Although animal models have significantly contributed to understanding chronic pain pathophysiology, the majority of research has focused on male rodents using testing procedures that produce sex difference data that do not align well with comparable clinical experiences. Additionally, the use of animal pain models presents ongoing ethical challenges demanding continuing refinement of preclinical methods. To this end, this study sought to test a quantitative allodynia assessment technique and associated statistical analysis in a modified graded nerve injury pain model with the aim to further examine sex differences in allodynia. Graded allodynia was established in male and female Sprague Dawley rats by altering the number of sutures placed around the sciatic nerve and quantified by the von Frey test. Linear mixed effects modeling regressed response on each fixed effect (sex, oestrus cycle, pain treatment). On comparison with other common von Frey assessment techniques, utilizing lower threshold filaments than those ordinarily tested, at 1 s intervals, appropriately and successfully investigated female mechanical allodynia, revealing significant sex and oestrus cycle difference across the graded allodynia that other common behavioral methods were unable to detect. Utilizing this different von Frey approach and graded allodynia model, a single suture inflicting less allodynia was sufficient to demonstrate exaggerated female mechanical allodynia throughout the phases of dioestrus and pro-oestrus. Refining the von Frey testing method, statistical analysis technique and the use of a graded model of chronic pain, allowed for examination of the influences on female mechanical nociception that other von Frey methods cannot provide.Lauren Nicotra, Jonathan Tuke, Peter M. Grace, Paul E. Rolan and Mark R. Hutchinso

The power of effective study design in animal Experimentation: Exploring the statistical and ethical implications of asking multiple questions of a data set

One of the chief advantages of using highly standardised biological models including model organisms is that multiple variables can be precisely controlled so that the variable of interest is more easily studied. However, such an approach often obscures effects in sub-populations resulting from natural population heterogeneity. Efforts to expand our fundamental understanding of multiple sub-populations are in progress. However, such stratified or personalised approaches require fundamental modifications of our usual study designs that should be implemented in Brain, Behavior and Immunity (BBI) research going forward. Here we explore the statistical feasibility of asking multiple questions (including incorporating sex) within the same experimental cohort using statistical simulations of real data. We illustrate and discuss the large explosion in sample numbers necessary to detect effects with appropriate power for every additional question posed using the same data set. This exploration highlights the strong likelihood of type II errors (false negatives) for standard data and type I errors when dealing with complex genomic data, where studies are too under-powered to appropriately test these interactions. We show this power may differ for males and females in high throughput data sets such as RNA sequencing. We offer a rationale for the use of alternative experimental and statistical strategies based on interdisciplinary insights and discuss the realworld implications of increasing the complexities of our experimental designs, and the implications of not attempting to alter our experimental designs going forward.R.A. Ankeny, A.L. Whittaker, M. Ryan, J. Boer, M. Plebanski, J. Tuke, S.J. Spence

Admixture has obscured signals of historical hard sweeps in humans (advance online)

The role of natural selection in shaping biological diversity is an area of intense interest in modern biology. To date, studies of positive selection have primarily relied on genomic datasets from contemporary populations, which are susceptible to confounding factors associated with complex and often unknown aspects of population history. In particular, admixture between diverged populations can distort or hide prior selection events in modern genomes, though this process is not explicitly accounted for in most selection studies despite its apparent ubiquity in humans and other species. Through analyses of ancient and modern human genomes, we show that previously reported Holocene-era admixture has masked more than 50 historic hard sweeps in modern European genomes. Our results imply that this canonical mode of selection has probably b een underappreciated in the evolutionary history of humans and suggest that our current understanding of the tempo and mode of selection in natural populations may be inaccurat

Codeine-induced hyperalgesia and allodynia: investigating the role of glial activation

Chronic morphine therapy has been associated with paradoxically increased pain. Codeine is a widely used opioid, which is metabolized to morphine to elicit analgesia. Prolonged morphine exposure exacerbates pain by activating the innate immune toll-like receptor-4 (TLR4) in the central nervous system. In silico docking simulations indicate codeine also docks to MD2, an accessory protein for TLR4, suggesting potential to induce TLR4-dependent pain facilitation. We hypothesized codeine would cause TLR4-dependent hyperalgesia/allodynia that is disparate from its opioid receptor-dependent analgesic rank potency. Hyperalgesia and allodynia were assessed using hotplate and von Frey tests at days 0, 3 and 5 in mice receiving intraperitoneal equimolar codeine (21 mg kg(-1)), morphine (20 mg kg(-1)) or saline, twice daily. This experiment was repeated in animals with prior partial nerve injury and in TLR4 null mutant mice. Interventions with interleukin-1 receptor antagonist (IL-1RA) and glial-attenuating drug ibudilast were assessed. Analyses of glial activation markers (glial fibrillary acid protein and CD11b) in neuronal tissue were conducted at the completion of behavioural testing. Despite providing less acute analgesia (P=0.006), codeine induced similar hotplate hyperalgesia to equimolar morphine vs saline (-9.5 s, P<0.01 and -7.3 s, P<0.01, respectively), suggesting codeine does not rely upon conversion to morphine to increase pain sensitivity. This highlights the potential non-opioid receptor-dependent nature of codeine-enhanced pain sensitivity-although the involvement of other codeine metabolites cannot be ruled out. IL-1RA reversed codeine-induced hyperalgesia (P<0.001) and allodynia (P<0.001), and TLR4 knock-out protected against codeine-induced changes in pain sensitivity. Glial attenuation with ibudilast reversed codeine-induced allodynia (P<0.001), and thus could be investigated further as potential treatment for codeine-induced pain enhancement.J L Johnson, P E Rolan, M E Johnson, L Bobrovskaya, D B Williams, K Johnson, J Tuke, and M R Hutchinso