Refinement and revalidation of the Equine Ophthalmic Pain Scale: R-EOPS a new scale for ocular pain assessment in horses

This study addresses the refinement and revalidation of a composite pain scale that focuses on equine facial expressions and behavioural indicators as exhibitions of ophthalmic pain. This scale included only Behavioural and Facial and Ocular expression indicators and, compared to the first version of Equine Ophthalmic Pain Scale (EOPS), item descriptors and related ratings were changed. Thirteen horses with ocular diseases that required medical or surgical treatment were enroled (group P). In each animal, the refined EOPS (R-EOPS) was applied prior to any treatment (T0) and one week later (T7). The R-EOPS was applied twice, 7 days apart, to 16 healthy control horses (group C). Two 30-second videos were recorded each time to allow the retrospective analysis by eight observers. Inter-observer reliability of items was moderate or substantial (Krippendorff's alpha, Ka>0.40) while their intra-observer reliability was substantial or almost perfect for most items (Ka ≥0.61). Both inter- and intra-observer reliability of Total Score (TS) were however excellent (Intraclass Correlation Coefficients, ICC>0.75). The TS also showed good reproducibility (Kendall coefficient=0.786, ICC=0.684) and high consistency of its items (Cronbach's a=0.847). The comparison between groups as well as the sensitivity and specificity values supported the validity of the R-EOPS. In particular, for each extra point added to the TS, the risk of the horse having pain increased by more than two times (Odds Ratio=2.079, 95%CI=1.542-2.804; P<0.001). The Receiver Operating Characteristic analysis identified 6 as the threshold value of R-EOPS for discriminating horses with ocular pathology (sensitivity=83%, specificity=100%). This scale may be an effective tool for reliably assessing the pain level in horses with ophthalmic diseases and potentially guiding pain management although it still requires large-scale application and external validation

Development and preliminary validation of a pain scale for ophthalmic pain in horses: The Equine Ophthalmic Pain Scale (EOPS)

The purpose of this study was to describe the development and preliminary validation of a composite pain scale, called the Equine Ophthalmic Pain Scale (EOPS), to assess ocular pain in horses. Indicators associated with ocular pain were selected and classified as behavioural, physiological or ocular expressions. Eight horses diagnosed with ocular or adnexa diseases that required medical or surgical treatment were enrolled in the study (group P). The developed EOPS was applied at the baseline (T0) and 1 week later (T7). Moreover, the EOPS was applied twice, 1 week apart, to 15 healthy control horses (group C). Videos of 60-80 s duration of all assessments were retrospectively analysed by seven masked observers, who scored items included in the behavioural and ocular expression categories of the EOPS.The inter- and intra-observer reliability was excellent (intraclass correlation coefficients >0.75) for most of the scored items. Cronbach's alpha (0.76) indicated that the EOPS had good internal consistency. The total score (TS), calculated as the sum of all scores, differed between groups C and P at TO (P < 0.001) and reduced after medical/surgical treatment in group P (P = 0.017), indicating the responsiveness of the EOPS. Moreover, the area under the curve (AUC=0.918, 95% confidence interval = 0.815-1.000; P < 0.001) indicated that the EOPS was very accurate for distinguishing healthy from pathological animals. Sensitivity and specificity of EOPS to identify horses with ocular pathology (at the optimal cut-off, i.e. TS >= 7) were 81.3% and 100.0%, respectively. However, 'overall behaviour', 'position inside the box', 'ear movements' and 'head position' items as well as physiological parameters, showed sub-optimal reliability, consistency and/or item-total correlation, suggesting that there is still mom to improve this composite scale

Comparison of Mesh and Barbed Suture for Laparoscopic Nephrosplenic Space Ablation in Horses

Nephrosplenic space (NSS) ablation has been demonstrated to be an effective technique for prevention of left dorsal displacement of the large colon and multiple laparoscopic techniques, among which ablation with mesh or with a barbed suture, have been proposed. Our objective was to compare two laparoscopic techniques for closure of the NSS in twenty-eight horses diagnosed with nephrosplenic entrapment. Medical records of horses that had laparoscopic NSS ablation in two referral centers between 2017–2019 were retrieved. Duration of surgery, complications, and short- and long-term follow-up information were collected and compared. Costs were also calculated and compared. All horses met the inclusion criteria: 9 had NSS ablation with a mesh implant (group M), 19 with barbed suture material (group B). One horse in group B had recurrent colic after discharge. At control laparoscopy after 5 months, the NSS resulted in still not being ablated because of a failure of the suture. In group M, three horses had recurrent colic. One was successfully treated medically, one died of unknown causes and the third required a second laparoscopic suturing at 3–6 months because of failure of the mesh implant. The mean time of surgery and costs were lower in group B compared to group M. The barbed suture technique was faster, more cost-effective and had a lower complication rate than the mesh implant

Synthesis and Biological Evaluation of New Clofibrate Analogues as Potential PPARa Agonists

Clofibrate is a lipid-profile modifying agent belonging to the fibrate class of drugs. Fibrates are known to exhibit their beneficial effects by activating peroxisome proliferator-activated receptor-a (PPARa) and used in the treatment of dyslipidemia and atherosclerosis and for the prevention of heart failure. Hereby, the preparation of two new sets of clofibrate analogues, ethyl 2-(4-chlorophenoxy)-3-oxoalkanoates and ethyl 2-(4-chlorophenoxy)-3-hydroxyalkanoates is described starting from commercially available 3-oxoalkanoates in fair to good yields. Treatment of 3-oxoalkanoates with SO2Cl2 yielded the corresponding 2-chloro-3-oxoalkanoates, that were then converted into 2-(4- chlorophenoxy)-3-oxoalkanoates by reacting with sodium or caesium 4-chlorophenate. Reduction of the keto group with NaBH4 afforded the corresponding 2-(4-chlorophenoxy)-3-hydroxyalkanoates in very high yields and with variable diastereoselectivity. Biological evaluation of the compounds was performed by a transactivation assay in a transiently transfected monkey kidney fibroblast cell line. The newly synthesised clofibrate analogues failed to show noticeable levels of PPAR activation at concentrations where clofibrate showed an evident activity, suggesting that the structural modifications caused the loss of PPAR activity

Inhibition of cyclooxygenase-1 does not reduce mortality in post-ischemic stroke rats

Background: Ischemic stroke is one of the leading causes of mortality and morbidity. The currently available non-invasive therapeutic options are not sufficiently efficacious. Post-ischemic brain is characterized by a prominent inflammatory response. Little is known about the involvement of cyclooxygenase (COX)-1 in the pathophysiology of ischemic stroke. Objective: This study was undertaken to examine the effects of a highly selective COX-1 inhibitor – mofezolac – on clinical outcomes and brain inflammatory markers in post-stroke rats. Methods: Stroke was induced by subjecting rats to permanent middle cerebral artery occlusion (MCAO). Control rats underwent a sham surgery. Rats were treated with mofezolac (50 mg/kg, intraperitoneally [ip]) once daily for 14 days. Control animals were treated with vehicle. Body temperature (BT), neurological score (NS) and cumulative mortality were monitored at different time points. At the end of the experiment, rats were euthanized and three brain regions (hypothalamus, hippocampus and frontal cortex) were extracted. Levels of interleukin (IL)-6, prostaglandin (PG)E2 and tumor necrosis factor (TNF)-α in these brain regions were determined by ELISA kits. Results: BT, NS and cumulative mortality were all significantly higher in post-MCAO rats than in sham-operated rats, irrespective of the treatment given. BT, NS and mortality rate did not differ significantly between mofezolac-treated and vehicle-treated sham-operated animals. BT was significantly lower in mofezolac-treated as compared to vehicle-treated post-MCAO rats. Mofezolac did not significantly alter NS in post-MCAO rats at any time-point. Cumulative 14-day mortality was non-significantly higher in mofezolac-treated as compared to vehicle-treated post-MCAO rats (48 % vs. 21 %, respectively; P = 0.184). Mostly, IL-6 and TNF-α levels did not differ between post-MCAO and sham-operated rats and were not affected by mofezolac treatment. In contrast, mofezolac significantly decreased PGE2 levels in post-MCAO rats’ brains. Conclusion: Overall, these results suggest that chronic treatment with the selective COX-1 inhibitor mofezolac did not reduce morbidity or mortality in post-stroke rats

Single-dose extended-toxicity preclinical study on novel radiotracer formulations for use in the diagnosis of neuroendocrine tumor and neurodegenerative disorders

The use of [18F]F-DOPA as an imaging agent for positron emission tomography (PET) studies of neurodegeneration and tumor detection is hindered by the chemical instability in its injectate formulation, which is also accompanied by injection-site reactions. Here we investigated novel F-DOPA lactate-based formulations that minimize formulation toxicity without compromising chemical stability. In vivo tolerability study on ND2 (lactate/Na2EDTA/FDOPA) and ND3 (acetic acid/F-DOPA) formulations in rats (i.v./i.m. 0.025-5 mg/kg) and mice (i.v. 50 mg/kg) was performed. Single-dose extended-toxicity study was performed in rats and mice in order to evaluate the acute and long-lasting response of the animals to the novel formulations. Toxicity related to ionizing radiation was not investigated because the stable isotope [19F]F-DOPA was used instead of [18F]F-DOPA which is the fluorine radioisotope that decays by b+ emission. The i.m. injection of ND3 (5 mg/kg) and ND3-vehicle to rats caused a local reaction characterized by up-regulation of the autophagic Lc3 and Bnip3, apoptotic Caspase 3, 8 and 9, mitochondrial Pgc1α, and inflammatory Mapk3, Cgrp, TNFα genes. Mapk3 and Pgc1α were also up-regulated in the ND2 (5 mg/kg)-treated muscles. The i.v. injection of ND3 (5 mg/kg) caused a reduction of body weight in rats, after 14 days of follow-up; the ND3 (50 mg/kg) and the ND3-vehicle caused a loss of body weight of-17.8 ± 1% and-12.3 ± 2% vs controls, respectively, in mice. No effects were observed following the administration of ND2 and the ND2-vehicle in rats and mice. The F-DOPA lactate/Na2EDTA-based formulation is better tolerated than the acetic acid-based formulation

Key Research Priorities for Factories of the Future—Part I: Missions

This chapter investigates research priorities for factories of the future by adopting an approach based on mission-oriented policies to support manufacturing innovation. Missions are challenging from a scientific and technological point of view and, at the same time, are addressing problems and providing results that are understandable by common people. Missions are based on clear targets that can help mitigating grand challenges. Based on the results of the Italian Flagship Project Factories of the Future, this chapter proposes seven missions while identifying the societal impact, the technological and industrial challenges, and the barriers to be overcome. These missions cover topics such as circular economy, rapid and sustainable industrialisation, robotic assistant, factories for personalised medicine, internet of actions, factories close to the people, and turning ideas into products. The accomplishment of missions asks for the support of a proper research environment in terms of infrastructures to test and demonstrate the results to a wide public. Research infrastructures together with funding mechanisms will be better addressed in the next chapter of this book