Evaluation of viscoelastic coagulation monitoring parameters and fibrinogen concentrations in healthy dogs undergoing stifle arthroscopy and tibial plateau leveling osteotomy.

OBJECTIVE: Evaluation of viscoelastic parameters along with Hct percentages, platelet numbers, and fibrinogen concentrations in healthy dogs undergoing elective stifle arthroscopy and tibial plateau leveling osteotomy (TPLO). These results determine if dogs are in a normal, hypocoagulable, or hypercoagulable state at 4 different time points. ANIMALS: Prospective study of 37 client-owned dogs enrolled between February 2, 2022, and November 10, 2022. METHODS: All dogs received a standardized anesthetic protocol. Patients enrolled in the study underwent stifle arthroscopy using a 3-portal technique with a TPLO performed by board-certified surgeons. Viscoelastic testing, Hct percentage, fibrinogen concentration, and platelet numbers were measured preoperatively, immediately postoperatively, 24 hours postoperatively, and 14 days postoperatively. RESULTS: Stifle arthroscopy and TPLO surgery was not associated with significant effects on viscoelastic coagulation monitoring and fibrinogen concentrations in healthy dogs when comparing the time points. CLINICAL RELEVANCE: Humans have a high risk of thrombotic events when undergoing elective orthopedic surgery and are often placed on prophylactic antithrombotic medication prior to surgery. Viscoelastic coagulation monitoring in dogs undergoing similar orthopedic procedures has been evaluated in veterinary medicine, but the effects on platelets and fibrinogen concentrations from similar orthopedic procedures and uniform anesthesia protocols have not been documented. Cranial cruciate ligament insufficiency is the most common orthopedic injury in dogs. Treatment requires elective surgical correction for the best results and improved long-term prognosis. The findings of this study suggest that similar preoperative prophylactic antithrombotic intervention is not needed for canine patients.

Alfaxalone provides reliable sedation for intradermal allergen testing in dogs and does not significantly affect results when compared to dexmedetomidine.

BACKGROUND: Dexmedetomidine (Dexmedetomidine hydrochloride-Dexdomitor; Zoetis) is the preferred sedative used for canine intradermal allergen testing (IDT) in the United States. Alfaxalone (Alfaxan Multidose; Jurox Animal Health) is a neuroactive steroid, and its effect on sedation and allergen reactivity scores is unknown. HYPOTHESIS/OBJECTIVES: We hypothesised that alfaxalone would produce an adequate level of sedation with fewer cardiovascular adverse effects and would not affect allergen reactivity scores or histamine wheal size compared to dexmedetomidine. MATERIALS AND METHODS: Twenty client-owned dogs were included in two groups: 10 atopic and 10 nonatopic. In a randomised, controlled, blinded, cross-over design all dogs underwent two modified IDT, 1-4 weeks apart, using intravenous dexmedetomidine (2.87-5.22 µg/kg) or alfaxalone (1.8-2.4 mg/kg). Anaesthetic parameters and sedation level were recorded over 25 min using a validated canine sedation scale described by Grint et al. (Small Anim Pract, 2009, 50, 62). Simultaneously, both objective and subjective reactivity scores were measured in technical triplicates at 10, 15 and 20 min. The modified IDT included eight allergens, histamine-positive and saline-negative controls. RESULTS: Alfaxalone produced a significantly higher sedation score across all time points (p < 0.05). All objective scores were significantly correlated to the corresponding subjective scores (Spearman R = 0.859, p < 0.0001). Sedative used did not significantly affect subjective allergen scores for nine atopic dogs (p > 0.05, 15 min). Sedative used did not affect the diameter of objective scores for individual allergens and histamine wheals (p > 0.05, 15 min). CONCLUSIONS AND CLINICAL RELEVANCE: Intravascular alfaxalone is an alternative sedative for IDT in dogs. Alfaxalone may be preferred to dexmedetomidine in some clinical scenarios as a result of having fewer cardiovascular adverse effects.

Metabolic remodeling precedes mTORC1-mediated cardiac hypertrophy.

RATIONALE: The nutrient sensing mechanistic target of rapamycin complex 1 (mTORC1) and its primary inhibitor, tuberin (TSC2), are cues for the development of cardiac hypertrophy. The phenotype of mTORC1 induced hypertrophy is unknown. OBJECTIVE: To examine the impact of sustained mTORC1 activation on metabolism, function, and structure of the adult heart. METHODS AND RESULTS: We developed a mouse model of inducible, cardiac-specific sustained mTORC1 activation (mTORC1iSA) through deletion of Tsc2. Prior to hypertrophy, rates of glucose uptake and oxidation, as well as protein and enzymatic activity of glucose 6-phosphate isomerase (GPI) were decreased, while intracellular levels of glucose 6-phosphate (G6P) were increased. Subsequently, hypertrophy developed. Transcript levels of the fetal gene program and pathways of exercise-induced hypertrophy increased, while hypertrophy did not progress to heart failure. We therefore examined the hearts of wild-type mice subjected to voluntary physical activity and observed early changes in GPI, followed by hypertrophy. Rapamycin prevented these changes in both models. CONCLUSION: Activation of mTORC1 in the adult heart triggers the development of a non-specific form of hypertrophy which is preceded by changes in cardiac glucose metabolism.

Comparison of methohexital and propofol as induction agents for evaluation of laryngeal function in healthy dogs.

OBJECTIVE: To determine the influence of propofol or methohexital, with and without doxapram, on the examination of laryngeal function in dogs. STUDY DESIGN: Experimental study. ANIMALS: Forty healthy dogs randomly assigned to 4 groups: propofol with saline (n = 10), propofol with doxapram (n = 10), methohexital with saline (n = 10), or methohexital with doxapram (n = 10). METHODS: Propofol and methohexital were administered to effect. Investigators examined laryngeal function (initial) simultaneously with video laryngoscopy. Doxapram or saline was administered, and laryngeal function was reevaluated (second). Laryngeal motion, quality of laryngeal exposure, and the degree of swallowing, laryngospasm, and jaw tone were scored at each evaluation. Adverse events were recorded. Initial and second videos were evaluated by a masked observer, and still images obtained from both evaluations were evaluated for change in rima glottidis size by 2 masked observers. RESULTS: Administration of doxapram and saline was delayed with propofol (P = .001). Laryngeal function did not differ between dogs receiving propofol or methohexital, irrespective of doxapram administration. Doxapram improved breathing scores in both groups (P < .001). Jaw tone increased with propofol during the second evaluation (P = .049). Swallowing was more prevalent at initial examination (P = .020). Methohexital resulted in an increased heart rate (P < .001) compared with propofol. Twenty-five percent of dogs receiving methohexital developed seizure-like activity (n = 5/20). CONCLUSION: Evaluation of laryngeal function did not differ between healthy dogs anesthetized with propofol or methohexital. Methohexital provided shorter examination times with less jaw tone but was associated with adverse events. CLINICAL SIGNIFICANCE: This study provides evidence to recommend propofol over methohexital as an induction agent for laryngeal function examination.

Markers of autophagy are downregulated in failing human heart after mechanical unloading.

BACKGROUND: Autophagy is a molecular process that breaks down damaged cellular organelles and yields amino acids for de novo protein synthesis or energy provision. Mechanical unloading with a left ventricular assist device (LVAD) decreases the energy demand of the failing human heart. We tested the hypothesis that LVAD support reverses activation of autophagy. METHODS AND RESULTS: Paired biopsy samples of left ventricular myocardium were obtained from 9 patients with idiopathic dilated cardiomyopathy (mean duration of LVAD support, 214 days) at the time of implantation and explantation of the LVAD. Transcript and protein levels of markers and mediators of autophagy and apoptosis were measured by quantitative reverse-transcription polymerase chain reaction and Western blotting. TUNEL assays, C9 immunohistochemistry, and 20S proteasome activity assays were also performed. Mechanical unloading significantly decreased mRNA transcript levels of Beclin-1, autophagy-related gene 5 (Atg5), and microtubule-associated protein-1 light chain-3 (MAP1-LC3 or LC3; P<0.02). Protein levels of Beclin-1, Atg5-Atg12 conjugate, and LC3-II were also significantly reduced after LVAD support (P<0.05). A significant increase in 20S proteasome activity was observed with unloading, in parallel to the decrease in autophagic markers. Although BNIP3 and the ratio of activated caspase 3 to procaspase 3 increased after LVAD support, Bcl-2 and TUNEL-positive nuclei were not significantly different between samples. CONCLUSIONS: Mechanical unloading of the failing human heart decreases markers of autophagy. These findings suggest that autophagy may be an adaptive mechanism in the failing heart, and this phenomenon is attenuated by LVAD support.

Induction of antioxidant gene expression in a mouse model of ischemic cardiomyopathy is dependent on reactive oxygen species.

Ischemia and reperfusion (I/R) are characterized by oxidative stress as well as changes in the antioxidant enzymes of the heart. However, little is known about the transcriptional regulation of myocardial antioxidant enzymes in repetitive I/R and hibernating myocardium. In a mouse model of ischemic cardiomyopathy induced by repetitive I/R, we postulated that induction of antioxidant gene expression was dependent on reactive oxygen species (ROS). Repetitive closed-chest I/R (15 min) was performed daily in C57/BL6 mice and in mice overexpressing extracellular superoxide dismutase (EC-SOD). Antioxidant enzyme expression was measured at 3, 5, 7, and 28 days of repetitive I/R as well as 15 and 30 days after discontinuation of I/R. In order to determine whether ROS directly modulates antioxidant gene expression, transcript levels were measured in cardiomyocytes exposed to hydrogen peroxide. Repetitive I/R caused an early and sustained increase in glutathione peroxidase (GPX) transcript levels, while heme oxygenase-1 (HO-1) expression increased only after 7 days of repetitive I/R. Overexpression of EC-SOD prevented the upregulation of GPX and HO-1 transcript levels by repetitive I/R, suggesting that both genes are regulated by ROS. However, while HO-1 transcript levels increased in cardiomyocytes exposed to hydrogen peroxide, oxidative stress failed to induce the expression of GPX implying that ROS regulates GPX transcript levels only indirectly in repetitive I/R. In conclusion, repetitive I/R was associated with an early upregulation of GPX expression as well as a delayed increase of HO-1 transcript levels in the heart. The induction of both antioxidant genes was dependent on ROS, suggesting that alterations in redox balance mediate not only tissue injury but also components of "programmed cell survival" in hibernating myocardium.

Downregulation of peroxisome proliferator-activated receptor-alpha gene expression in a mouse model of ischemic cardiomyopathy is dependent on reactive oxygen species and prevents lipotoxicity.

BACKGROUND: The peroxisome proliferators-activated receptor-alpha (PPARalpha), a transcription factor that modulates fatty acid metabolism, regulates substrate preference in the heart. Although in acute ischemia there is a switch in substrate preference from fatty acids to glucose, metabolic gene expression in repetitive ischemia is not well described. In a mouse model of ischemic cardiomyopathy induced by repetitive ischemia/reperfusion (I/R), we postulated that downregulation of PPARalpha is regulated by reactive oxygen species and is necessary for maintaining contractile function in the heart. METHODS AND RESULTS: Repetitive closed-chest I/R (15 minutes) was performed daily in C57/BL6 mice, mice overexpressing extracellular superoxide dismutase, and mice treated with the PPARalpha agonist-WY-14,643. Echocardiography, histology, and candidate gene expression were measured at 3, 5, 7, and 28 days of repetitive I/R and 15 and 30 days after discontinuation of I/R. Repetitive I/R was associated with a downregulation of PPARalpha-regulated genes and both myosin heavy chain isoform transcript levels, which was reversible on discontinuation of I/R. Overexpression of EC-SOD prevented the downregulation of PPARalpha-regulated genes and myosin iso-genes by repetitive I/R. Furthermore, reactivation of PPARalpha in mice exposed to repetitive I/R worsened contractile function, induced microinfarctions, and increased intramyocardial triglyceride deposition, features suggestive of cardiac lipotoxicity. CONCLUSIONS: Metabolic and myosin isoform gene expression in repetitive I/R is mediated by reactive oxygen species. Furthermore, we suggest that downregulation of PPARalpha in repetitive I/R is an adaptive mechanism that is able to prevent lipotoxicity in the ischemic myocardium.