Effect of weight-adjusted intermediate-dose versus fixed-dose prophylactic anticoagulation with low-molecular-weight heparin on venous thromboembolism among noncritically and critically ill patients with COVID-19: the COVI-DOSE trial, a multicenter, randomised, open-label, phase 4 trial.

Background: Venous thromboembolism is a major complication of coronavirus disease 2019 (COVID-19). We hypothesized that a weight-adjusted intermediate dose of anticoagulation may decrease the risk of venous thromboembolism COVID-19 patients. Methods: In this multicenter, randomised, open-label, phase 4, superiority trial with blinded adjudication of outcomes, we randomly assigned adult patients hospitalised in 20 French centers and presenting with acute respiratory SARS-CoV-2. Eligible patients were randomly assigned (1:1 ratio) to receive an intermediate weight-adjusted prophylactic dose or a fixed-dose of subcutaneous low-molecular-weight heparin during the hospital stay. The primary outcome corresponded to symptomatic deep-vein thrombosis (fatal) pulmonary embolism during hospitalization (COVI-DOSE ClinicalTrials.gov number: NCT04373707). Findings: Between May 2020, and April 2021, 1000 patients underwent randomisation in medical wards (noncritically ill) (80.1%) and intensive care units (critically ill) (19.9%); 502 patients were assigned to receive a weight-adjusted intermediate dose, and 498 received fixed-dose thromboprophylaxis. Symptomatic venous thromboembolism occurred in 6 of 502 patients (1.2%) in the weight-adjusted dose group and in 10 of 498 patients (2.1%) in the fixed-dose group (subdistribution hazard ratio, 0.59; 95% CI, 0.22-1.63; P = 0.31). There was a twofold increased risk of major or clinically relevant nonmajor bleeding: 5.9% in the weight-adjusted dose group and 3.1% in the fixed-dose group (P = 0.034). Interpretation: In the COVI-DOSE trial, the observed rate of thromboembolic events was lower than expected in patients hospitalized for COVID-19 infection, and the study was unable to show a significant difference in the risk of venous thromboembolism between the two low-molecular-weight-heparin regimens. Funding: French Ministry of Health, CAPNET, Grand-Est Region, Grand-Nancy Métropole.

Mining the equine gut metagenome: poorly-characterized taxa associated with cardiovascular fitness in endurance athletes.

Emerging evidence indicates that the gut microbiome contributes to endurance exercise performance. Still, the extent of its functional and metabolic potential remains unknown. Using elite endurance horses as a model system for exercise responsiveness, we built an integrated horse gut gene catalog comprising ~25 million unique genes and 372 metagenome-assembled genomes. This catalog represents 4179 genera spanning 95 phyla and functional capacities primed to exploit energy from dietary, microbial, and host resources. The holo-omics approach shows that gut microbiomes enriched in Lachnospiraceae taxa are negatively associated with cardiovascular capacity. Conversely, more complex and functionally diverse microbiomes are associated with higher glucose concentrations and reduced accumulation of long-chain acylcarnitines and non-esterified fatty acids in plasma, suggesting increased ß-oxidation capacity in the mitochondria. In line with this hypothesis, more fit athletes show upregulation of mitochondrial-related genes involved in energy metabolism, biogenesis, and Ca2+ cytosolic transport, all of which are necessary to improve aerobic work power, spare glycogen usage, and enhance cardiovascular capacity. The results identify an associative link between endurance performance and gut microbiome composition and gene function, laying the basis for nutritional interventions that could benefit horse athletes.

Anatomical distribution of Toxoplasma gondii in naturally and experimentally infected lambs.

Consumption of raw or undercooked meat containing Toxoplasma gondii tissue cysts is one of the main sources of infection for humans worldwide. Among the various species intended for human consumption, sheep appear to be a high risk for human infection. The present study focused on the detailed anatomical distribution of Toxoplasma gondii in naturally and experimentally infected lambs using fresh and frozen samples of various pieces of meat, from a public health perspective. The first objective was to rank the edible parts intended for human consumption according to the detectable parasite burden by real-time PCR targeting the 529-bp repeated element. The second objective was to evaluate the impact of freezing by comparing the detection efficiency of the quantitative PCR between fresh and frozen tissues, as imports of lamb carcasses/cuts may arrive frozen or chilled. The highest estimated parasite loads were observed in skeletal muscles, and more particularly in edible portions such as quadriceps femoris muscle, intercostal muscles, deltoid muscle and diaphragm, with a significant difference in detectable parasite burden between fresh and frozen samples (p < 0.0001) or natural and experimental infection (p < 0.0001). Thoracic and pelvic limbs (3278-1048 parasites/g muscle) were ranked at the top of the list. Toxoplasma gondii DNA was detected in all the edible parts of lamb studied. These results suggest that lamb meat represents a risk for consumers. Further investigations are needed in order to confirm these differences in larger numbers of animals and in different breeds.

Title: Distribution anatomique de Toxoplasma gondii chez des agneaux infectés naturellement et expérimentalement. Abstract: La consommation de viande crue ou insuffisamment cuite contenant des kystes tissulaires de Toxoplasma gondii est l'une des principales sources d'infection pour l'homme dans le monde. Parmi les différentes espèces destinées à la consommation humaine, le mouton apparaît à haut risque d'infection humaine. La présente étude s'est concentrée sur une distribution anatomique détaillée de Toxoplasma gondii chez des agneaux infectés naturellement et expérimentalement à l'aide d'échantillons frais et congelés de divers morceaux de viande, du point de vue de la santé publique. Classer les parties comestibles destinées à la consommation humaine, selon la charge parasitaire détectable par une PCR en temps réel ciblant l'élément répété de 529 pb était un premier objectif. Un second objectif était d'évaluer l'impact de la congélation en comparant l'efficacité de détection de la PCR quantitative entre les tissus frais et congelés, car les importations de carcasses/coupes d'agneau peuvent arriver congelées ou réfrigérées. Les charges parasitaires estimées les plus élevées ont été observées dans les muscles squelettiques et plus particulièrement dans les parties comestibles telles que le quadriceps fémoral, les muscles intercostaux, le deltoïde et le diaphragme avec une différence significative de charge parasitaire détectable entre les échantillons frais et congelés (p < 0,0001) ou l'infection naturelle et expérimentale (p < 0,0001). Les membres thoraciques et pelviens (3278 à 1048 parasites/g de muscle) ont été classés en tête de liste. L'ADN de T. gondii a été détecté dans toutes les parties comestibles étudiées de l'agneau. Ces résultats suggèrent que l'agneau représente un risque pour les consommateurs. Des investigations supplémentaires doivent être effectuées afin de confirmer les différences mentionnées ci-dessus chez plus d'animaux et dans différentes races.

Progression to a severe form of COVID-19 among patients with chronic respiratory diseases.

RATIONALE: Viral respiratory infections, including SARS-CoV-2 infection, can trigger respiratory symptoms among patients suffering from chronic respiratory diseases, leading to exacerbations and hospitalizations. Despite the tropism of SARS-CoV-2 into the respiratory tract, chronic respiratory diseases do not seem to be risk factors for severe forms of COVID-19. OBJECTIVES: To assess whether hospitalized patients for COVID-19 with chronic respiratory diseases were at lower risk of developing a severe form than other patients. METHODS: This French study included patients admitted to hospital in COVID-19 ward, suffering from a SARS-CoV-2 infection, diagnosed on RT-PCR or chest computed tomography associated with clinical symptoms, from March 15 to June 30, 2020. Ambulatory patients who were tested in the emergency department and patients with severe hypoxaemia requiring intensive care were not included. All data were collected from electronic medical records up to discharge of the patient. MAIN RESULTS: 617 patients were included: 125 with a chronic respiratory disease, mainly chronic obstructive pulmonary disease (45%) and asthma (30%). The percentage of patients scoring 6 or higher on the WHO Clinical Progression Scale during hospital stay was lower in patients with chronic respiratory disease compared to those without chronic respiratory disease (21.6% versus 31.3%, respectively, p = 0.03). Among patients with chronic respiratory disease, temperature above 38 °C on admission (OR 16.88 (95% CI 4.01-71.00)), lymphopenia (OR 5.08 (1.25-20.72)), CPAP therapy (OR 4.46 (1.04-19.17)) and age (OR 1.09 (1.02-1.16)) were associated with an increased risk to reach a score of 6 or above. CONCLUSIONS: Hospital admissions in COVID-19 ward of patients suffering from chronic respiratory diseases are at lower risk of developing a severe form of COVID- 19, especially in patients with chronic obstructive pulmonary disease or asthma. Prospective studies would confirm our results and allow to better organize the follow-up of these patients in a pandemic period.

Understanding the Holobiont: Crosstalk Between Gut Microbiota and Mitochondria During Long Exercise in Horse.

Endurance exercise has a dramatic impact on the functionality of mitochondria and on the composition of the intestinal microbiome, but the mechanisms regulating the crosstalk between these two components are still largely unknown. Here, we sampled 20 elite horses before and after an endurance race and used blood transcriptome, blood metabolome and fecal microbiome to describe the gut-mitochondria crosstalk. A subset of mitochondria-related differentially expressed genes involved in pathways such as energy metabolism, oxidative stress and inflammation was discovered and then shown to be associated with butyrate-producing bacteria of the Lachnospiraceae family, especially Eubacterium. The mechanisms involved were not fully understood, but through the action of their metabolites likely acted on PPARγ, the FRX-CREB axis and their downstream targets to delay the onset of hypoglycemia, inflammation and extend running time. Our results also suggested that circulating free fatty acids may act not merely as fuel but drive mitochondrial inflammatory responses triggered by the translocation of gut bacterial polysaccharides following endurance. Targeting the gut-mitochondria axis therefore appears to be a potential strategy to enhance athletic performance.

Thyrohyoideus muscle innervation in the horse.

OBJECTIVE: To describe the innervation of the thyrohyoideus (TH) muscle and to confirm our findings with stimulation of first cervical (C1) nerve branches. STUDY DESIGN: Ex vivo phase 1 and clinical phase 2. ANIMALS: Fourteen head and neck specimens and 17 client-owned horses. METHODS: In phase 1, the cranial nerve (CN) XII and the C1 nerve were dissected with their branches in 20 dissections were performed on 14 specimens (6 left and right side and 8 only left or right) Anatomy was noted. Samples of nerve bifurcations were collected for histological confirmation of anatomical findings. First cervical nerve branches were stimulated in horses undergoing cervical nerve graft to treat laryngeal hemiplegia. RESULTS: The nerve innervating the TH muscle arose directly from the C1 nerve in 17 of 20 dissections, from an anastomotic branch between CN XII and the C1 nerve in two of 20 dissections, and from the C1 nerve and the anastomotic branch in one of 20 dissections. No direct connection between the TH muscle and CN XII was found. Histological examination revealed that the anastomosis was composed of C1 nerve fibers passing over to CN XII. First cervical stimulation resulted in TH muscle contraction in 16 of 17 horses. CONCLUSIONS: The innervation of the TH muscle originated from the C1 nerve according to dissection, histological, and conduction studies, with variation in the branching pattern. CLINICAL SIGNIFICANCE: Care should be taken to preserve the C1 nerve during prosthetic laryngoplasty. The surgical technique for C1 nerve grafts should be reconsidered in light of these findings, along with new options to treat dorsal displacement of the soft palate..

Publisher Correction: Unraveling the effects of the gut microbiota composition and function on horse endurance physiology.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Unraveling the effects of the gut microbiota composition and function on horse endurance physiology.

An integrated analysis of gut microbiota, blood biochemical and metabolome in 52 endurance horses was performed. Clustering by gut microbiota revealed the existence of two communities mainly driven by diet as host properties showed little effect. Community 1 presented lower richness and diversity, but higher dominance and rarity of species, including some pathobionts. Moreover, its microbiota composition was tightly linked to host blood metabolites related to lipid metabolism and glycolysis at basal time. Despite the lower fiber intake, community type 1 appeared more specialized to produce acetate as a mean of maintaining the energy supply as glucose concentrations fell during the race. On the other hand, community type 2 showed an enrichment of fibrolytic and cellulolytic bacteria as well as anaerobic fungi, coupled to a higher production of propionate and butyrate. The higher butyrate proportion in community 2 was not associated with protective effects on telomere lengths but could have ameliorated mucosal inflammation and oxidative status. The gut microbiota was neither associated with the blood biochemical markers nor metabolome during the endurance race, and did not provide a biomarker for race ranking or risk of failure to finish the race.

A First Step Toward Unraveling the Energy Metabolism in Endurance Horses: Comparison of Plasma Nuclear Magnetic Resonance Metabolomic Profiles Before and After Different Endurance Race Distances.

Endurance racing places high demands on energy metabolism pathways. Metabolomics can be used to investigate biochemical responses to endurance exercise in humans, laboratory animals, and horses. Although endurance horses have previously been assessed in the field (i.e., during races) using broad-window Nuclear Magnetic Resonance metabolomics, these studies included several different race locations, race distances, age classes, and race statuses (finisher or elimination). The present NMR metabolomics study focused on 40 endurance horses racing in three race categories over 90, 120, or 160 km. The three races took place in the same location. Given that energy metabolism is closely related to exercise intensity and duration (and therefore distance covered), the study's objective was to determine whether the metabolic pathways recruited during the race varied as a function of the total ride distance. For each horse, a plasma sample was collected the day before the race, and another was collected at the end of the race. Sixteen, 15, and 9 horses raced over 90, 120, and 160 km, respectively. Proton NMR spectra (500 MHz) were acquired for these 80 plasma samples. After processing, the spectra were divided into bins representing the NMR variables and then classified using orthogonal projection on latent structure models supervised by the sampling time (pre- or post-race) or the distance covered. The models revealed that the post-race metabolomic profiles are associated to the total ride distance groups. By combining biochemical assay results and NMR data in multiblock models, we further showed that enzymatic activities and metabolites are significantly associated to the race category. In the highest race category (160 km), there appears to be a metabolic switch from carbohydrate consumption to lipid consumption in order to maintain glycaemia. Furthermore, signs of protein breakdown were more apparent in the longest race category. The metabolic shift seen in the different racing categories could be related to a mixture of three important factors that are the ride distance, the training status and the inherited endurance capacity of the various horses competing.

Genotype imputation accuracy in multiple equine breeds from medium- to high-density genotypes.

Genotype imputation is now a key component of genomic analyses as it increases the density of available genotypes within a population. However, many factors can influence imputation accuracy. The aim of this study was to assess and compare the accuracy of imputation of high-density genotypes (Affymetrix Axiom Equine genotyping array, 670,806 SNPs) from two moderate-density genotypes (Illumina Equine SNP50 BeadChip, 54,602 SNPs and Illumina Equine SNP70 BeadChip, 65,157 SNPs), using single-breed or multiple-breed reference sets. Genotypes were available from five groups of horse breeds: Arab (AR, 1,207 horses), Trotteur Français (TF, 979 horses), Selle Français (SF, 1,979 horses), Anglo-Arab (AA, 229 horses) and various foreign sport horses (FH, 209 horses). The proportions of horses genotyped with the high-density (HD) chip in each breed group were 10% in AA, 15% in AR and FH, 30% in TF and 57% in SF. A validation set consisting of one-third of the horses genotyped with the HD chip was formed and their genotypes deleted. Two imputation strategies were compared, one in which the reference population consisted only of horses from the same breed group as in the validation set, and another with horses from all breed groups. For the first strategy, concordance rates (CRs) ranged from 97.8% (AR) to 99.0% (TF) and correlations (r²) from 0.94 (AR) to 0.99 (TF). For the second strategy, CR ranged from 97.4% (AR) to 98.9% (TF) and r² from 0.93 (AR) to 0.99 (TF). Overall, the results show a small advantage of within-breed imputation compared with multi-breed imputation. Adding horses from different breed groups to the reference population does not improve the accuracy of imputation. Imputation provides an accurate means of combining data sets from different genotyping platforms, now necessary with the increasing use of the recently developed Affymetrix Axiom Equine genotyping array.

Anatomy of the vestibulum esophagi and surgical implications during prosthetic laryngoplasty in horses.

OBJECTIVE: To describe the anatomy of the entry to the equine esophagus (vestibulum esophagi) and to assess the risk of penetrating its adventitia and/or lumen during laryngoplasty. STUDY DESIGN: Ex vivo cadaveric study. SAMPLE POPULATION: Five isolated equine larynges and 39 equine head and neck specimens. METHODS: The anatomy of the vestibulum esophagi was studied by dissection of 5 cadaver specimens. Then, a bilateral laryngoplasty was performed, including 5 suture placements through the muscular processes, caudal, rostral, and sagittal, with straight and curved needles. Two of the 3 surgeons performing the implantations were unaware of the goals of the study. Suture positions and iatrogenic trauma to the lumen and/or adventitia of the vestibulum esophagi were identified during dissection of the specimens. Risk factors for penetrating the adventitia were evaluated with a multivariate regression model. RESULTS: The vestibulum esophagi spans between both wings of the thyroid cartilage over the entire width of the larynx, covering the rostral spine (arcuate crest) of the arytenoid cartilages. It is covered by the thyropharyngeus and cricopharyngeus muscles. Masked surgeons were associated with a significantly higher number of adventitia penetrations (72%) compared to the nonmasked surgeon (9%). The lumen of the vestibulum esophagi was penetrated in 4.6% of suture placements and only by the 2 masked surgeons. CONCLUSION: Penetration of the adventitia was more common when surgeons were unaware of the anatomical extent of the vestibulum esophagi. CLINICAL SIGNIFICANCE: Anatomical knowledge of the extent of the vestibulum esophagi reduces the risk of penetrating its lumen or adventitia during suture placement on the muscular process of the arytenoid cartilage.

Agreement between Electrocardiogram and Heart Rate Meter Is Low for the Measurement of Heart Rate Variability during Exercise in Young Endurance Horses.

Analysis of the heart rate variability (HRV) gains more and more importance in the assessment of training practice and welfare in equine industry. It relies on mathematical analyses of reliably and accurately measured variations in successive inter-beat intervals, measured as RR intervals. Nowadays, the RR intervals can be obtained through two different techniques: a heart rate meter (HRM) or an electrocardiogram (ECG). The agreement and reliability of these devices has not been fully assessed, especially for recordings during exercise. The purpose of this study was to assess the agreement of two commercially available devices using the two mentioned techniques (HRM vs ECG) for HRV analysis during a standardized exercise test. Simultaneous recordings obtained during light exercise and during canter with both devices were available for 36 horses. Data were compared using a Bland-Altman analysis and the Lin's coefficient. The agreement between the assessed HRV measures from the data obtained from the ECG and HRM was acceptable only for the mean RR interval and the mean heart rate. For the other studied measures (SDNN, root mean square of successive differences, SD1, SD2, low frequency, high frequency), the agreement between the devices was too poor for them to be considered as interchangeable in these recording conditions. The agreement tended also to be worse when speed of the exercise increased. Therefore, it is necessary to be careful when interpreting and comparing results of HRV analysis during exercise, as the results will depend upon recording devices. Furthermore, corrections and data processing included in the software of the devices affect largely the output used in the subsequent HRV analysis; this must be considered in the choice of the device.

Endurance Exercise Ability in the Horse: A Trait with Complex Polygenic Determinism.

Endurance horses are able to run at more than 20 km/h for 160 km (in bouts of 30-40 km). This level of performance is based on intense aerobic metabolism, effective body heat dissipation and the ability to endure painful exercise. The known heritabilities of endurance performance and exercise-related physiological traits in Arabian horses suggest that adaptation to extreme endurance exercise is influenced by genetic factors. The objective of the present genome-wide association study (GWAS) was to identify single nucleotide polymorphisms (SNPs) related to endurance racing performance in 597 Arabian horses. The performance traits studied were the total race distance, average race speed and finishing status (qualified, eliminated or retired). We used three mixed models that included a fixed allele or genotype effect and a random, polygenic effect. Quantile-quantile plots were acceptable, and the regression coefficients for actual vs. expected log10p-values ranged from 0.865 to 1.055. The GWAS revealed five significant quantitative trait loci (QTL) corresponding to 6 SNPs on chromosomes 6, 1, 7, 16, and 29 (two SNPs) with corrected p-values from 1.7 × 10-6 to 1.8 × 10-5. Annotation of these 5 QTL revealed two genes: sortilin-related VPS10-domain-containing receptor 3 (SORCS3) on chromosome 1 is involved in protein trafficking, and solute carrier family 39 member 12 (SLC39A12) on chromosome 29 is active in zinc transport and cell homeostasis. These two coding genes could be involved in neuronal tissues (CNS). The other QTL on chromosomes 6, 7, and 16 may be involved in the regulation of the gene expression through non-coding RNAs, CpG islands and transcription factor binding sites. On chromosome 6, a new candidate equine long non-coding RNA (KCNQ1OT1 ortholog: opposite antisense transcript 1 of potassium voltage-gated channel subfamily Q member 1 gene) was predicted in silico and validated by RT-qPCR in primary cultures of equine myoblasts and fibroblasts. This lncRNA could be one element of the cardiac rhythm regulation. Our GWAS revealed that equine performance during endurance races is a complex polygenic trait, and is partially governed by at least 5 QTL: two coding genes involved in neuronal tissues and three other loci with many regulatory functions such as slowing down heart rate.

Understanding the response to endurance exercise using a systems biology approach: combining blood metabolomics, transcriptomics and miRNomics in horses.

BACKGROUND: Endurance exercise in horses requires adaptive processes involving physiological, biochemical, and cognitive-behavioral responses in an attempt to regain homeostasis. We hypothesized that the identification of the relationships between blood metabolome, transcriptome, and miRNome during endurance exercise in horses could provide significant insights into the molecular response to endurance exercise. For this reason, the serum metabolome and whole-blood transcriptome and miRNome data were obtained from ten horses before and after a 160 km endurance competition. RESULTS: We obtained a global regulatory network based on 11 unique metabolites, 263 metabolic genes and 5 miRNAs whose expression was significantly altered at T1 (post- endurance competition) relative to T0 (baseline, pre-endurance competition). This network provided new insights into the cross talk between the distinct molecular pathways (e.g. energy and oxygen sensing, oxidative stress, and inflammation) that were not detectable when analyzing single metabolites or transcripts alone. Single metabolites and transcripts were carrying out multiple roles and thus sharing several biochemical pathways. Using a regulatory impact factor metric analysis, this regulatory network was further confirmed at the transcription factor and miRNA levels. In an extended cohort of 31 independent animals, multiple factor analysis confirmed the strong associations between lactate, methylene derivatives, miR-21-5p, miR-16-5p, let-7 family and genes that coded proteins involved in metabolic reactions primarily related to energy, ubiquitin proteasome and lipopolysaccharide immune responses after the endurance competition. Multiple factor analysis also identified potential biomarkers at T0 for an increased likelihood for failure to finish an endurance competition. CONCLUSIONS: To the best of our knowledge, the present study is the first to provide a comprehensive and integrated overview of the metabolome, transcriptome, and miRNome co-regulatory networks that may have a key role in regulating the metabolic and immune response to endurance exercise in horses.

Feasibility, repeatability, and safety of ultrasound-guided stimulation of the first cervical nerve at the alar foramen in horses.

OBJECTIVE To develop and assess the feasibility, repeatability, and safety of an ultrasound-guided technique to stimulate the first cervical nerve (FCN) at the level of the alar foramen of the atlas of horses. ANIMALS 4 equine cadavers and 6 clinically normal Standardbreds. PROCEDURES In each cadaver, the FCN pathway was determined by dissection, and any anastomosis between the first and second cervical nerves was identified. Subsequently, each of 6 live horses underwent a bilateral ultrasound-guided stimulation of the FCN at the alar foramen 3 times at 3-week intervals. After each procedure, horses were examined daily for 5 days. RESULTS In each cadaver, the FCN passed through the alar foramen; a communicating branch between the FCN and the accessory nerve and anastomoses between the ventral branches of the FCN and second cervical nerve were identified. The anastomoses were located in the upper third of the FCN pathway between the wing of the atlas and the nerve's entry in the omohyoideus muscle. Successful ultrasound-guided electrical stimulation was confirmed by twitching of the ipsilateral omohyoideus muscle in all 6 live horses; this finding was observed bilaterally during each of the 3 experimental sessions. No complications developed at the site of stimulation. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that ultrasound-guided stimulation of the FCN at the alar foramen appears to be a safe and straightforward procedure in horses. The procedure may have potential for use in horses with naturally occurring recurrent laryngeal neuropathy to assess reinnervation after FCN transplantation or nerve-muscle pedicle implantation in the cricoarytenoideus dorsalis muscle.

Effects of Age, Exercise Duration, and Test Conditions on Heart Rate Variability in Young Endurance Horses.

Although cardiac recovery is an important criterion for ranking horses in endurance competitions, heart rate variability (HRV) has hardly ever been studied in the context of this equestrian discipline. In the present study, we sought to determine whether HRV is affected by parameters such as age, exercise duration and test site. Accordingly, HRV might be used to select endurance horses with the fastest cardiac recovery. The main objective of the present study was to determine the effects of age, exercise duration, and test site on HRV variables at rest and during exercise and recovery in young Arabian endurance horses. Over a 3-year period, 77 young Arabian horses aged 4-6 years performed one or more exercise tests (consisting of a warm-up, cantering at 22 km.h(-1)and a final 500 m gallop at full speed) at four different sites. Beat-to-beat RR intervals were continuously recorded and then analyzed (using a time-frequency approach) to determine the instantaneous HRV components before, during and after the test. At rest, the root-mean-square of successive differences in RR intervals (RMSSD) was higher in the 4-year-olds (54.4 ± 14.5 ms) than in the 5-or 6-year-olds (44.9 ± 15.5 and 49.1 ± 11.7 ms, respectively). During the first 15 min of exercise (period T), the heart rate (HR) and RMSSD decreased with age. In 6-year-olds, RMSSD decreased as the exercise duration increased (T: 3.0 ± 1.4 vs. 2T: 3.6 ± 2.2 vs. 3T: 2.8 ± 1.0). During recovery, RMSSD was negatively correlated with the cardiac recovery time (CRT) and the recovery heart rate (RHR; R = -0.56 and -0.53, respectively; p < 0.05). At rest and during exercise and recovery, RMSSD and several HRV variables differed significantly as a function of the test conditions. HRV in endurance horses appears to be strongly influenced by age and environmental factors (such as ambient temperature, ambient humidity, and track quality). Nevertheless, RMSSD can be used to select endurance horses with the fastest cardiac recovery.

Integrated mRNA and miRNA expression profiling in blood reveals candidate biomarkers associated with endurance exercise in the horse.

The adaptive response to extreme endurance exercise might involve transcriptional and translational regulation by microRNAs (miRNAs). Therefore, the objective of the present study was to perform an integrated analysis of the blood transcriptome and miRNome (using microarrays) in the horse before and after a 160 km endurance competition. A total of 2,453 differentially expressed genes and 167 differentially expressed microRNAs were identified when comparing pre- and post-ride samples. We used a hypergeometric test and its generalization to gain a better understanding of the biological functions regulated by the differentially expressed microRNA. In particular, 44 differentially expressed microRNAs putatively regulated a total of 351 depleted differentially expressed genes involved variously in glucose metabolism, fatty acid oxidation, mitochondrion biogenesis, and immune response pathways. In an independent validation set of animals, graphical Gaussian models confirmed that miR-21-5p, miR-181b-5p and miR-505-5p are candidate regulatory molecules for the adaptation to endurance exercise in the horse. To the best of our knowledge, the present study is the first to provide a comprehensive, integrated overview of the microRNA-mRNA co-regulation networks that may have a key role in controlling post-transcriptomic regulation during endurance exercise.

Impact of CB1 Receptor Deletion on Visual Responses and Organization of Primary Visual Cortex in Adult Mice.

PURPOSE: The endocannabinoids (eCBs) and their receptors are expressed in the cortex of developing animals where they act as a neuromodulating system during critical stages of brain development such as cell proliferation and migration, and axon guidance. Little is known on the impact of the cannabinoid system on cortical map formation and receptive field properties of cortical sensory neurons. The present study evaluates in vivo the functional organization of the primary visual cortex (V1) of mice lacking cannabinoid CB1R receptor (cnr1-/-). METHODS: Using optical imaging of intrinsic signals, azimuth, and elevation maps of cnr1-/- mice were compared with their wild-type littermates (cnr1+/+). RESULTS: Topographic maps were affected in mutant mice as they exhibited narrower visual field and changes in the shape of V1. CB1R exerted its action in an axis dependent manner as all changes were observed in the azimuth axis. Spatial frequency and contrast sensitivity were also compared between the two groups. Both properties were affected by the chronic lacking of CB1R as mutant mice exhibited a significantly lower contrast sensitivity as well as lower spatial frequency selectivity. CONCLUSIONS: Taken together, these results suggest an important role for CB1R in cortical map formation. Our results also clearly demonstrate the impact of CB1R in the development of visual properties of primary visual cortex neurons. Because psychoactive effects of cannabis consumption on visual experience are mediated mainly through CB1R, our results could possibly explain neuronal mechanisms involved in those perceptual changes.

Speed and Cardiac Recovery Variables Predict the Probability of Elimination in Equine Endurance Events.

Nearly 50% of the horses participating in endurance events are eliminated at a veterinary examination (a vet gate). Detecting unfit horses before a health problem occurs and treatment is required is a challenge for veterinarians but is essential for improving equine welfare. We hypothesized that it would be possible to detect unfit horses earlier in the event by measuring heart rate recovery variables. Hence, the objective of the present study was to compute logistic regressions of heart rate, cardiac recovery time and average speed data recorded at the previous vet gate (n-1) and thus predict the probability of elimination during successive phases (n and following) in endurance events. Speed and heart rate data were extracted from an electronic database of endurance events (80-160 km in length) organized in four countries. Overall, 39% of the horses that started an event were eliminated--mostly due to lameness (64%) or metabolic disorders (15%). For each vet gate, logistic regressions of explanatory variables (average speed, cardiac recovery time and heart rate measured at the previous vet gate) and categorical variables (age and/or event distance) were computed to estimate the probability of elimination. The predictive logistic regressions for vet gates 2 to 5 correctly classified between 62% and 86% of the eliminated horses. The robustness of these results was confirmed by high areas under the receiving operating characteristic curves (0.68-0.84). Overall, a horse has a 70% chance of being eliminated at the next gate if its cardiac recovery time is longer than 11 min at vet gate 1 or 2, or longer than 13 min at vet gates 3 or 4. Heart rate recovery and average speed variables measured at the previous vet gate(s) enabled us to predict elimination at the following vet gate. These variables should be checked at each veterinary examination, in order to detect unfit horses as early as possible. Our predictive method may help to improve equine welfare and ethical considerations in endurance events.

Energetics of endurance exercise in young horses determined by nuclear magnetic resonance metabolomics.

Long-term endurance exercise severely affects metabolism in both human and animal athletes resulting in serious risk of metabolic disorders during or after competition. Young horses (up to 6 years old) can compete in races up to 90 km despite limited scientific knowledge of energetic metabolism responses to long distance exercise in these animals. The hypothesis of this study was that there would be a strong effect of endurance exercise on the metabolomic profiles of young horses and that the energetic metabolism response in young horses would be different from that of more experienced horses. Metabolomic profiling is a powerful method that combines Nuclear Magnetic Resonance (NMR) spectrometry with supervised Orthogonal Projection on Latent Structure (OPLS) statistical analysis. (1)H-NMR spectra were obtained from plasma samples drawn from young horses (before and after competition). The spectra obtained before and after the race from the same horse (92 samples) were compared using OPLS. The statistical parameters showed the robustness of the model (R2Y = 0.947, Q2Y = 0.856 and cros-validated ANOVA p < 0.001). For confirmation of the predictive value of the model, a test set of 104 sample spectra were projected by the model, which provided perfect predictions as the area under the receiving-operator curve was 1. The metabolomic profile determined with the OPLS model showed that glycemia after the race was lower than glycemia before the race, despite the involvement of lipid and protein catabolism. An OPLS model was calculated to compare spectra obtained on plasma taken after the race from 6-year-old horses and from experienced horses (cross-validated ANOVA p < 0.001). The comparison of metabolomic profiles in young horses to those from experienced horses showed that experienced horses maintained their glycemia with higher levels of lactate and a decrease of plasma lipids after the race.