Artificial intelligence is a powerful technology with great potential to support veterinarians across many aspects of their multifaceted job. However, realizing this potential requires AI solutions that truly address specific pain points. Here we review 4 use cases in which AI has been applied to small animal veterinary medicine: image analysis, early disease detection, administration support, and disease surveillance. For each of these, we briefly present the current state of the technology and available AI applications. Results show that tangible value creation is heterogeneous across use cases and that availability of accurate and reliable data (eg, in the form of curated electronic health re- cords) is a major limiting factor.

BACKGROUND:Medication review type 3 (MR3) is a comprehensive and collaborative clinical service designed to optimize patient treatment. The BRussels ANTwerp Medication Review Quality score (BRANT-MERQS) assesses MR3 quality, incorporating 45 general and 4 project-specific criteria. Using all BRANT-MERQS criteria is laborious and time-intensive, necessitating the development of a more efficient, streamlined version that preserves reliability. AIM:The aim of this study was to create a reliable and more efficient scoring system to evaluate the quality of MR3. METHOD:A random but weighted subset of the BRANT-MERQS quality criteria was employed in repeated tests. This technique helped determine the required subsample size for accurate quality scores. The weighting was based on previously assessed importance of each criterion. RESULTS:Repeated sampling showed that a 10-criteria random subsample yielded dependable quality scores. The biased sampling approach (5-3-1 ratio, 5 for highly important criteria, 3 for moderately important criteria and 1 for less important criteria) offered improvements compared to an unbiased one. CONCLUSION:A more efficient MR3 quality evaluation was developed using the BRANT-MERQS scoring table. A 10 criteria biased subsample, representing one-third of the full assessment, ensures both time efficiency and consistency in quality assessment.

Background and Aims: Fluoroquinolones (FQ) have been linked to aortic aneurysms and dissections (AA/AD), resulting in an official warning. However, recent large-scale epidemiological studies have reported lack of FQ-AA/AD association. This study aimed to scrutinize FQ-AA/AD risk by implementing a combined epidemiological and experimental approach. Methods: Danish nationwide registers (2003-2021) were used for a nested case-control analysis. FQ-AA/AD risk was evaluated in a main and high-risk FDA cohort. Further, mortality and aortic interventions linked to FQ were investigated in patients with aortic disease. Additionally, ciprofloxacin (100 mg/kg/day, 2x2 weeks) was administered to wild-type, hypertensive or Marfan mice. Aortic diameters and pulse wave velocity (PWV) were measured longitudinally to investigate aortic remodelling. Results: The main cohort comprised 5.10 million individuals with 58,919 cases and 1,767,510 sampled controls. Compared with amoxicillin exposure. FQ exposure was not associated with increased AA/AD risk (30-day hazard ratios (HR) 1.00 [95% confidence intervals (CI): 0.74-1.34]; 90-day HR 1.07 [CI 0.94-1.22]; 1-year HR 0.95 [0.90-1.01]). In a high-risk cohort, there was no FQ-AA/AD association (30-day HR 0.83 [0.61-1.12]; 90-day HR 0.99 [0.86-1.15]; 1-year HR 0.97 [0.90-1.05]). In patients with aortic disease, FQ were not associated with increased aortic interventions or mortality (30-day HR 0.98 [0.79-1.22]; 90-day HR 1.06 [0.95-1.19]). Additionally, ciprofloxacin did not affect aortic diameters or PWV in wild type, hypertensive, and Marfan mice, while differences between models proved the sensitivity of the methodology. Conclusion: The data clearly do not support the current precautions and warnings pertaining to risks of aortopathies and FQ should not be discouraged when clinically indicated.

Background and Aims: Epidemiological data reveal a higher incidence of atherosclerosis in cancer survivors. In addition to its well-recognised cardiotoxicity, the chemotherapeutic agent doxorubicin (DOX) impairs endothelial function and increases arterial stiffness, which are key antecedents for atherosclerosis development. DOX may therefore accelerate atherosclerotic plaque formation, yet this has not been investigated so far. The current study therefore investigated whether DOX accelerates atherosclerotic plaque formation. Methods: 24 male and 24 female apolipoprotein E-knockout (ApoE-/-) mice (age 6 weeks) were fed a high-fat (HF) diet for 17 weeks. Starting at week 3, DOX (4 mg/kg) was administered intraperitoneally to half of the mice in each group once a week for 3 weeks. After 17 weeks, mice were humanely killed to assess the amount and the content of plaque in the thoracic aorta, proximal aorta and brachiocephalic artery. Results: DOX increased arterial stiffness in male and female mice after 2 weeks, validating our model. Overall, total plaque amount and plaque content did not differ in the thoracic aorta and brachiocephalic artery of DOX-treated mice while sex differences in plaque size were observed. Paradoxically, DOX treatment resulted in smaller plaques in female mice, but not male mice. Conclusions: In contrast to epidemiological reports, DOX does not seem to accelerate atherosclerotic plaque formation, although this study needs to be confirmed in a treatment study with shorter follow-up.

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Geconcerteerde Onderzoeksactie (GOA) grant 499 of the University of Antwerp Industrieel OnderzoeksFonds/Strategisch Basisonderzoek 498 (IOF/SBO) research grant of the University of Antwerp Background Atrial fibrosis is a substrate of atrial fibrillation (AF), and higher burdens of fibrosis are associated with resistance to therapy and worse prognosis. Currently, no therapies exist that target atrial fibrosis. JK07 is a long-acting neuregulin-fusion protein that has been shown to decrease ventricular fibrosis in models of heart failure through selective stimulation of the ErbB4 receptor. Purpose To test the ability of JK07 to reduce atrial fibrosis and AF inducibility in minipig model of deoxycorticosterone acetate (DOCA, an aldosterone agonist) induced hypertension. Methods 18 Aachener minipigs were randomized into 3 groups: control (CTRL), DOCA + Vehicle (DOCA+VEH) and DOCA+JK07. The control group did not undergo a therapeutic intervention. To induce hypertension and atrial fibrosis, pellets releasing 10 mg/kg DOCA over a period of 60 days were implanted in minipigs in the DOCA+VEH and DOCA+JK07 groups. The DOCA-implanted animals underwent weekly treatment with JK07 (0.3 mg/kg; DOCA + JK07) or its vehicle (DOCA + VEH), starting at the day of implantation (total of 9 administrations). After 60 days, arterial blood pressure was measured invasively and a decapolar catheter was placed in the right atrium. AF inducibility was tested by performing 50 burst pacing episodes and quantified as the percentage of successful episodes where an AF run ≥ 5 sec could be induced after the burst, out of the 50 attempts. Atrial fibrosis was quantified using ImageJ software on Masson trichrome staining of left atrial specimens, isolated after euthanizing the animals. Results Mean arterial pressure was significantly higher in the DOCA+VEH (142 ± 10 mmHg) and DOCA+JK07 (132 ± 15 mmHg) groups than in the CTRL group (105 ± 8 mmHg, p<0.01), without an effect of JK07. AF inducibility in the DOCA+VEH group was significantly higher than in the CTRL group (66/250 vs. 9/300, p<0.001), and the DOCA + JK07 group (66/250 vs. 8/300, p<0.001). Likewise, the degree of atrial fibrosis was significantly higher in the DOCA+VEH group compared to CTRL (14.18 ± 1.81 vs. 8.30 ± 2.52, p<0.001) and compared to DOCA+JK07 (14.18 ± 1.81 vs. 10.65 ± 1.59, p=0.0049). Conclusions JK07 prevents atrial fibrosis and AF inducibility in a porcine DOCA model. The effect of JK07 is unrelated to effects on blood pressure, but probably related to reduced atrial fibrogenesis.

Abstract Background Before patients have spontaneous AF, pathologic remodeling is often already extensively present in the atria, creating a substrate for AF. Currently, no therapies exist that target this atrial remodeling, that can be divided into electrical (ion channel alterations) and structural (fibrosis, dilatation) remodeling. JK07 is a long-acting neuregulin-fusion protein with preventive effects towards ventricular fibrosis in models of heart failure through selective stimulation of the ERBB4 receptor. Purpose To test the ability of JK07 to reduce atrial fibrosis and AF inducibility in the minipig model of deoxycorticosterone acetate (DOCA, an aldosterone agonist) induced hypertension. Methods 18 Aachener minipigs were randomized into 3 groups: control (CTRL), DOCA + Vehicle (DOCA) and DOCA+JK07. The control group did not undergo disease induction or therapeutic intervention. To induce hypertension and atrial fibrosis, pellets releasing 100 mg/kg DOCA over a period of 60 days were implanted in minipigs in the DOCA+VEH and DOCA+JK07 groups. The DOCA-implanted animals underwent weekly treatment with JK07 (0.3 mg/kg; DOCA + JK07) or vehicle control (DOCA), starting on the day of implantation (total of 9 administrations). After 60 days, arterial blood pressure was measured invasively and a decapolar catheter was placed in the right atrium. AF inducibility was tested by performing 50 burst pacing episodes and quantified as the percentage of "successful" episodes where an AF run ≥ 5 sec could be induced after the burst, out of the total attempts per group. Atrial fibrosis was quantified using ImageJ software on Masson trichrome staining of atrial specimens. Results Mean arterial pressure was significantly higher in the DOCA+VEH (142 ± 10 mmHg) and DOCA+JK07 (132 ± 15 mmHg) groups than in the CTRL group (105 ± 8 mmHg, p<0.01), without an effect of JK07. AF inducibility in the DOCA+VEH group was significantly higher than in the CTRL group (66/250 vs. 9/300, p<0.001), and the DOCA + JK07 group (66/250 vs. 8/300, p<0.001). The degree of fibrosis in the left atrial free wall was significantly higher in the DOCA group compared to CTRL (12.16 ± 5.06 vs. 5.14 ± 1.57, p=0.0089) and compared to DOCA+JK07 (12.16 ± 5.06 vs. 5.47 ± 3.69, p=0.026). Similar results were also found in the right atrial free wall and left and right atrial appendage. Conclusions ERBB4 stimulation prevents atrial fibrotic remodeling and AF inducibility in a porcine DOCA model. The effect of JK07 is unrelated to effects on blood pressure, but probably related to reduced atrial fibrogenesis.ERBB4 stimulation prevents AF inductionERBB4 stimulation prevents fibrosis

An important substrate of atrial fibrillation (AF) is atrial fibrosis, and a higher burden of fibrosis correlates with worse prognosis and therapy resistance. Currently, no therapies exist that target atrial fibrosis. JK07 is a long-acting neuregulin-fusion protein that activates the ERBB4 receptor, which mitigates ventricular fibrosis in models of heart failure.

INTRODUCTION:A safety pharmacology study detects and evaluates potential side effects of a new drug on physiological function at therapeutic levels and above and, in most cases, prior to the initiation of clinical trials. The aim of this study was to investigate the effects of environmental and biological factors on resting heart rate (HR), a representative cardiac parameter in cardiovascular safety pharmacology.METHODS:Over twenty years, 143 dogs (Beagles, Labradors and mongrels) received implanted telemetry transmitters to measure aortic pressure (AP), left ventricular pressure (LVP), Electrocardiogram (ECG) and body temperature. Throughout the 7-h period of data collection, data were continuously recorded without drug treatment and included the range of HRs resulting from spontaneous physiological changes. Statistics and visualizations were calculated using R and Spotfire.RESULTS:Beagles had a higher HR than the mongrels, while Labradors had a lower HR than mongrels. Labradors were found to have a sex-based difference in HR, with females having a higher HR. A higher HR was observed in young animals of all breeds when they were in contact with humans. The cage system affected the HR of Labradors and mongrels more than Beagles. Larger dogs (e.g. Labrador) have a lower HR than smaller dogs (Beagles). Animals that are younger were found to have more HR variability and have a higher HR than older animals. In addition, older animals reacted less to the application period and human interaction than younger animals. The HR response of animals inside a cage system may depend on the cage system in which they were bred. A familiar cage system typically has less impact on HR.DISCUSSION:This retrospective data base evaluation has demonstrated the impact of environmental and biological factors on cardiovascular parameters in the context of performing safety pharmacology studies. Breed, sex, age and the type of cage system used affected, at least in some cases, the HR and its variability. They should therefore be carefully considered when designing safety pharmacology studies to have the highest possible test sensitivity.

Background and Aims : RIPK1 (receptor-interacting serine/threonine-protein kinase 1) enzymatic activity drives both apoptosis and necroptosis, a regulated form of necrosis. Because necroptosis is involved in necrotic core development in atherosclerotic plaques, we investigated the effect of a RIPK1S25D/S25D mutation, which prevents the activation of RIPK1 kinase, on atherogenesis in ApoE-/- mice. Furthermore, we pharmacologically inhibited RIPK1 kinase activity by administering GSK'547 to ApoE-/- Fbn1C1039G+/- mice, a mouse model of advanced atherosclerosis.Methods: ApoE-/- Ripk1+/+ (n=16) and ApoE-/- Ripk1S25D/S25D (n=12) mice were fed a Western-type diet (WD) for 16 weeks to induce plaque formation. ApoE-/- Fbn1C1039G+/- mice received WD supplemented with GSK'547 (10 mg/kg BW/day, n=12-13/group) for 20 weeks to evaluate the effect of pharmacological RIPK1 kinase inhibition on atherogenesis.Results: After 16 weeks WD, atherosclerotic plaques of ApoE-/- Ripk1S25D/S25D mice were significantly larger as compared to ApoE-/- Ripk1+/+ mice (167±34 vs. 69±18 103 μm2, P=0.01). Absolute cell numbers (350±34 vs. 154±33 nuclei) and deposition of glycosaminoglycans (Alcian blue: 31±6 vs. 14±4%, P=0.023) were increased in plaques from ApoE-/- Ripk1S25D/S25D mice while macrophage content (Mac3: 2.3±0.4 vs. 9.8±2.4%, P=0.012) was significantly decreased. Plaque apoptosis was not different between both groups. In contrast, pharmacological inhibition of RIPK1 kinase with GSK'547 in ApoE-/- Fbn1C1039G+/-mice did not significantly alter plaque size after 20 weeks WD, but induced apoptosis (TUNEL: 136±20 vs. 62±9 cells/mm2, P=0.004).Conclusions: Inhibition of RIPK1 kinase activity in ApoE-/- Ripk1S25D/S25D mice accelerated plaque progression, but this effect was not observed after pharmacological inhibition with GSK'547 in ApoE-/- Fbn1C1039G+/- mice. Background and Aims : RIPK1 (receptor-interacting serine/threonine-protein kinase 1) enzymatic activity drives both apoptosis and necroptosis, a regulated form of necrosis. Because necroptosis is involved in necrotic core development in atherosclerotic plaques, we investigated the effect of a RIPK1S25D/S25D mutation, which prevents the activation of RIPK1 kinase, on atherogenesis in ApoE-/- mice. Furthermore, we pharmacologically inhibited RIPK1 kinase activity by administering GSK'547 to ApoE-/- Fbn1C1039G+/- mice, a mouse model of advanced atherosclerosis. Methods: ApoE-/- Ripk1+/+ (n=16) and ApoE-/- Ripk1S25D/S25D (n=12) mice were fed a Western-type diet (WD) for 16 weeks to induce plaque formation. ApoE-/- Fbn1C1039G+/- mice received WD supplemented with GSK'547 (10 mg/kg BW/day, n=12-13/group) for 20 weeks to evaluate the effect of pharmacological RIPK1 kinase inhibition on atherogenesis. Results: After 16 weeks WD, atherosclerotic plaques of ApoE-/- Ripk1S25D/S25D mice were significantly larger as compared to ApoE-/- Ripk1+/+ mice (167±34 vs. 69±18 103 μm2, P=0.01). Absolute cell numbers (350±34 vs. 154±33 nuclei) and deposition of glycosaminoglycans (Alcian blue: 31±6 vs. 14±4%, P=0.023) were increased in plaques from ApoE-/- Ripk1S25D/S25D mice while macrophage content (Mac3: 2.3±0.4 vs. 9.8±2.4%, P=0.012) was significantly decreased. Plaque apoptosis was not different between both groups. In contrast, pharmacological inhibition of RIPK1 kinase with GSK'547 in ApoE-/- Fbn1C1039G+/-mice did not significantly alter plaque size after 20 weeks WD, but induced apoptosis (TUNEL: 136±20 vs. 62±9 cells/mm2, P=0.004). Conclusions: Inhibition of RIPK1 kinase activity in ApoE-/- Ripk1S25D/S25D mice accelerated plaque progression, but this effect was not observed after pharmacological inhibition with GSK'547 in ApoE-/- Fbn1C1039G+/- mice.

The aim of this study was to derive a model to predict the risk of dogs developing chronic kidney disease (CKD) using data from electronic health records (EHR) collected during routine veterinary practice. Data from 57,402 dogs were included in the study. Two thirds of the EHRs were used to build the model, which included feature selection and identification of the optimal neural network type and architecture. The remaining unseen EHRs were used to evaluate model performance. The final model was a recurrent neural network with 6 features (creatinine, blood urea nitrogen, urine specific gravity, urine protein, weight, age). Identifying CKD at the time of diagnosis, the model displayed a sensitivity of 91.4% and a specificity of 97.2%. When predicting future risk of CKD, model sensitivity was 68.8% at 1 year, and 44.8% 2 years before diagnosis. Positive predictive value (PPV) varied between 15 and 23% and was influenced by the age of the patient, while the negative predictive value remained above 99% under all tested conditions. While the modest PPV limits its use as a stand-alone diagnostic screening tool, high specificity and NPV make the model particularly effective at identifying patients that will not go on to develop CKD.

Background Cardiovascular disease remains the leading cause of death in chronic kidney disease (CKD) patients, especially in those undergoing dialysis and kidney transplant surgery. CKD patients are at high risk of developing arterial media calcifications (AMC) and arterial stiffness. We hypothesized that investigation of disease progression at an early stage could provide novel insights in understanding AMC etiology. Methods An adenine diet was administered to male Wistar rats to induce AMC. Rats were sacrificed after 2, 4 and 8 weeks. AMC was measured by assessment of aortic calcium and visualized using histology. Arterial stiffness was measured in vivo by ultrasound and ex vivo by applying cyclic stretch of physiological magnitude on isolated arterial segments, allowing us to generate the corresponding pressure-diameter loops. Further, ex vivo arterial reactivity was assessed in organ baths at 2 and 4 weeks to investigate early alterations in biomechanics/cellular functionality. Results CKD rats showed a time-dependent increase in aortic calcium which was confirmed on histology. Accordingly, ex vivo arterial stiffness progressively worsened. Pressure-diameter loops showed a gradual loss of arterial compliance in CKD rats. Additionally, viscoelastic properties of isolated arterial segments were altered in CKD rats. Furthermore, after 2 and 4 weeks of adenine treatment, a progressive loss in basal, nitric oxide (NO) levels was observed, which was linked to an increased vessel tonus and translates into an increasing viscous modulus. Conclusions Our observations indicate that AMC-related vascular alterations develop early after CKD induction prior to media calcifications being present. Preventive action, related to restoration of NO bioavailability, might combat AMC development.

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): Fonds voor Wetenschappelijk Onderzoek (FWO) Introduction Necroptosis is a form of regulated necrosis mediated by the kinase activity of receptor-interacting serine/threonine-protein kinase 1 (RIPK1) and executed by RIPK3 and mixed lineage kinase domain like pseudokinase (MLKL). Because necroptosis is involved in necrotic core development in atherosclerotic plaques, we investigated the effect of a RIPK1 S25D/S25D mutation, which prevents the activation of RIPK1 kinase, on atherogenesis in ApoE-/- mice. Furthermore, we pharmacologically inhibited RIPK1 kinase activity by administering GSK’547 to ApoE-/- Fbn1 C1039G+/- mice, a mouse model of advanced atherosclerosis. Purpose We aimed to study the effect of RIPK1 kinase inhibition on atherogenesis through a genetic and a pharmacological approach in ApoE-/- and ApoE-/- Fbn1 C1039G+/- mice, respectively. Methods ApoE-/- and ApoE-/- Fbn1 C1039G+/- mice were fed a Western-type diet (WD) for up to 24 weeks and plaque necroptosis was confirmed. ApoE-/- Ripk1+/+ (n=16) and ApoE-/- Ripk1 S25D/S25D (n=12) mice were fed a WD for 16 weeks to induce plaque formation. ApoE-/- Fbn1 C1039G+/- mice received WD supplemented with GSK’547 (10 mg/kg BW/day, n=12-13/group) for 20 weeks to evaluate the effect of pharmacological RIPK1 kinase inhibition on atherogenesis. Results After 16 weeks WD, atherosclerotic plaques of ApoE-/- Ripk1 S25D/S25D mice were significantly larger as compared to ApoE-/- Ripk1+/+ mice (167±34 vs. 69±18 103 µm2, P=0.01). Absolute cell numbers (350±34 vs. 154±33 nuclei) and deposition of glycosaminoglycans (Alcian blue: 31±6 vs. 14±4%, P=0.023) were increased in plaques from ApoE-/- Ripk1 S25D/S25D mice while macrophage content (Mac3: 2.3±0.4 vs. 9.8±2.4%, P=0.012) was significantly decreased. Increased deposition of glycosaminoglycans was also observed in the vessel wall of ApoE-/- Ripk1 S25D/S25D mice (Alcian blue: 4.3±1.5 vs. 1.4±0.4 %, P=0.04). Together with increased internal elastic lamina area and increased vessel wall thickness, this points towards hypertrophic positive vascular remodeling in ApoE-/- Ripk1 S25D/S25D mice. Plaque apoptosis was not different between both groups. In contrast, pharmacological inhibition of RIPK1 kinase with GSK’547 in ApoE-/- Fbn1 C1039G+/- mice did not significantly alter plaque size and vessel properties after 20 weeks WD, but induced plaque apoptosis (TUNEL: 124±17 vs. 68±8 cells/mm2, P=0.018; cleaved caspase 3: 0.49±0.13 vs. 0.14±0.04 %; p=0.021). Conclusion Inhibition of RIPK1 kinase activity in ApoE-/- Ripk1 S25D/S25D mice accelerated plaque progression and induced positive vascular remodeling. These effects were not observed after pharmacological inhibition with GSK’547 in ApoE-/- Fbn1 C1039G+/- mice. However, a switch to apoptosis was observed after GSK’547 treatment, stressing the complex involvement of RIPK1 in cell survival and death.

Introduction: Atrial fibrillation (AF) results from electrical and structural remodeling of the atria, in which inflammation and fibrosis play an important role. Current therapy is limited to antiarrhythmic drugs and ablations, but does not target the structural problem. Recent studies showed that neuregulin-1 (NRG1), an epidermal growth factor family member, has anti-fibrotic and anti-inflammatory effects in the myocardium. Purpose: To test the effects of JK07, a NRG1 antibody fusion comprising an ERBB3 antagonistic antibody which selectively signals through ERBB4 preferentially over ERBB3, on atrial fibrosis and AF inducibility. Methods: Atrial samples were harvested from male rats (Wistar Han, 10 weeks old), cut into small pieces (1-2mm 2 ) and kept in low serum medium in the presence or absence of JK07 (5nM). Col1a1 and Col3a1 mRNA was quantified after 24-72 hours. AF inducibility was tested in a first AF model in which male mice (C57BL/6N, 12-15 weeks old) were treated with angiotensin-II (Ang-II, 4 weeks, osmotic mini-pumps, 3000 ng/kg/min), and in a second AF model in which mice were fed with a high fat diet (HFD, 8 weeks, 60% Kcal fat) inducing severe weight gain (56±3% increase compared to 23±4% with regular chow). In both models, AF inducibility was tested by 5 runs of programmed electrical stimulation (PES) with a trans-jugular octapolar catheter. AF inducibility (% mice inducible by ≥3 PES-runs) and duration of PES-induced AF (AF duration) were recorded. Mice were randomized for treatment with vehicle or JK07 (2x/week, 1mg/kg, IV, n=5-7/group). Results: In cultured atrial samples, Col1a1 and Col3a1 mRNA expression gradually increased up to 2-3 fold over 3 days. JK07 robustly attenuated this effect by 59±17% (p<0.05). In mice, both Ang-II and HFD significantly increased AF inducibility and AF duration. In Ang-II mice, JK07 attenuated AF inducibility (from 57% to 20%) and AF duration (from 33.3 ± 15.1 to 1.5 ± 1s). In HFD mice, JK07 significantly attenuated AF inducibility (from 57% to 0%) and AF duration (from 10.9±3.2s to 0.76±0.5s, p<0.05). Conclusions: These results show anti-fibrotic effects by selective ERBB4 stimulation with JK07 in atrial tissue in vitro, together with AF-preventive effects in two unrelated mouse models.

The increasing attention for the dog-owner relationship combined with advances in nutrition and veterinary care have made wellbeing a focal point for dog owners, veterinarians, and dog product and service providers. While canine wellbeing can be quantified by survey-based quality of life instruments like those used in human healthcare, there are currently few instruments available that can do this reliably and at scale. Here we report the development and initial validation of a canine quality of life instrument specifically designed to quantify wellbeing in the general dog population. The instrument is based on a simple 32-question survey and includes 5 daytime domains (energetic, mobile, relaxed, happy, sociable) and 3 mealtime domains (relaxed, interested and satisfied). It captures specific health-related aspects as well as more general wellbeing aspects and, in an initial sample of 2813 dogs, already provides useful insights on canine wellbeing. We believe that data collection at scale with this instrument will help bring optimal wellbeing to the dogs we care for.

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): Research Foundation - Flanders (Brussels, BE): ==> [DUTCH]: "Fonds voor Wetenschappelijk Onderzoek (FWO) - Vlaanderen" Introduction The pulsatile character of blood flow in the central arteries results in a local transmission of energy to the arterial wall, acting on both the viscous and elastic components of the arterial wall. The amplitude of the "pulsatile load" is known to modulate the mechanosensitive properties of blood vessels. Purpose To evaluate the behavior of both the viscous and elastic components of the mouse aorta under altered pulsatile loading. Moreover, we aimed to assess regional differences in (arterial) tissue stiffness, as well as the contribution of vascular smooth muscle cell (VSMC) contractility under altered pulsatile conditions. Methods Aortic segments from C57Bl6 (n=9) mice were mounted in a Rodent Oscillatory Set-up for Arterial Compliance (ROTSAC), and subjected to high frequency (10 Hz) cyclic stretch at alternating loads (i.e. 20 to 100 mmHg with mean pressures ranging from 80 to 200 mmHg). Diastolic and systolic diameter, compliance, and the Peterson elastic modulus (Ep), as a measure of aortic stiffness, were determined. Viscous modulus (EV) was extracted from pressure-diameter tracings by eliminating loop hysteresis. Afterwards, the elastic modulus (EE) was calculated as the slope of the resulting pressure-diameter tracing: (P_elastic)= (P_total) - (EV * (dD/dt)) To assess regional differences in response to changes in pulsatile load, the most proximal aortic region (i.e., thoracic ascending aorta) was compared to a more distal aortic region (i.e., abdominal infrarenal aorta). Results Increasing pulse pressure from 40 to 100 mmHg decreased arterial stiffness (298 ± 9 vs. 270 ± 9 mmHg; p < 0.0001). Analyzing viscoelasticity revealed that increasing pulsatile load from 40 to 100 mmHg decreased EV (0.24 ± 0.02 vs. 0.21 ± 0.02 mmHg.s/mm; p < 0.01) but not EE. 50 mM KCl-induced VSMC contraction increased vessel stiffness (298 ± 9 mmHg vs. 395 ± 12 mmHg; p < 0.0001). This effect was pronounced at low pulse pressures (20 and 40 mmHg) while at higher pulse pressures (60, 80 and 100 mmHg) contraction-induced stiffness did not occur. Interestingly, the distal region of the aorta was stiffer than the proximal region (367 ± 60 vs. 289 ± 36 mmHg; p < 0.01) and had a higher EV (0.77 ± 0.2 vs. 0.21 ± 0.05 mmHg.s/mm; p < 0.0001). Increasing pulsatile load from 40 to 100 mmHg decreased the EV from distal aortic tissue more than the EV of tissue from the proximal region (-0.26 ± 0.02 vs. -0.06 ± 0.007 mmHg.s/mm). Conclusion High pulsatile load decreased arterial stiffness by decreasing the viscous mechanical properties. This effect was more pronounced in the distal region of the aorta.

Background and Aims : Gasdermin D (Gsdmd) is a key executioner of NLRP3 inflammasome- and caspase 1-dependent pyroptotic cell death. Recently, expression of Gsdmd and caspase 1-mediated cleavage of Gsdmd was described in atherosclerotic plaques. Plaque rupture is associated with strong immunoreactivity for caspase 1, suggesting that Gsdmd-mediated pyroptosis and inflammation are involved in plaque destabilization. Therefore, we aimed to study the effects of Gsdmd deficiency on advanced plaques in ApoE-/- mice.Methods: ApoE-/- Gsdmd-/- (n=16) and ApoE-/- Gsdmd+/+ (n=18) mice were fed a Western-type diet (WD) for 16 weeks to induce plaque formation. BMDMs were isolated from Gsdmd-/- and Gsdmd+/+ mice and treated with LPS followed by nigericin or ATP to characterize Gsdmd-dependent cell death and inflammation in vitro.Results: Atherosclerotic plaques in the brachiocephalic artery of WD-fed ApoE-/- Gsdmd-/-mice were significantly smaller as compared to controls (114±18 vs. 186±16 103 μm2, P=0.006). Moreover, plaques of ApoE-/- Gsdmd-/- mice showed features of increased plaque stability, such as a decrease in necrotic core area (19±4 vs. 37±7 103 μm2, P=0.03) and an increase in the vascular smooth muscle cell/macrophage ratio (αSMA/Mac3 ratio: 1.6±0.3 vs. 0.7±0.1, P=0.01). Interestingly, a significant increase of TUNEL positive cells was observed in plaques of ApoE-/- Gsdmd-/- mice as compared to controls (141±25 vs. 62±8 cells/mm2, P=0.005), suggesting a switch to apoptosis. This switch from pyroptotic to apoptotic cell death was also confirmed in vitro in Gsdmd-/- BMDM after LPS/nigericin and LPS/ATP treatment.Conclusions: Gsdmd deficiency delays atherogenesis but induces a switch to apoptosis in ApoE-/- mice. Background and Aims : Gasdermin D (Gsdmd) is a key executioner of NLRP3 inflammasome- and caspase 1-dependent pyroptotic cell death. Recently, expression of Gsdmd and caspase 1-mediated cleavage of Gsdmd was described in atherosclerotic plaques. Plaque rupture is associated with strong immunoreactivity for caspase 1, suggesting that Gsdmd-mediated pyroptosis and inflammation are involved in plaque destabilization. Therefore, we aimed to study the effects of Gsdmd deficiency on advanced plaques in ApoE-/- mice. Methods: ApoE-/- Gsdmd-/- (n=16) and ApoE-/- Gsdmd+/+ (n=18) mice were fed a Western-type diet (WD) for 16 weeks to induce plaque formation. BMDMs were isolated from Gsdmd-/- and Gsdmd+/+ mice and treated with LPS followed by nigericin or ATP to characterize Gsdmd-dependent cell death and inflammation in vitro. Results: Atherosclerotic plaques in the brachiocephalic artery of WD-fed ApoE-/- Gsdmd-/-mice were significantly smaller as compared to controls (114±18 vs. 186±16 103 μm2, P=0.006). Moreover, plaques of ApoE-/- Gsdmd-/- mice showed features of increased plaque stability, such as a decrease in necrotic core area (19±4 vs. 37±7 103 μm2, P=0.03) and an increase in the vascular smooth muscle cell/macrophage ratio (αSMA/Mac3 ratio: 1.6±0.3 vs. 0.7±0.1, P=0.01). Interestingly, a significant increase of TUNEL positive cells was observed in plaques of ApoE-/- Gsdmd-/- mice as compared to controls (141±25 vs. 62±8 cells/mm2, P=0.005), suggesting a switch to apoptosis. This switch from pyroptotic to apoptotic cell death was also confirmed in vitro in Gsdmd-/- BMDM after LPS/nigericin and LPS/ATP treatment. Conclusions: Gsdmd deficiency delays atherogenesis but induces a switch to apoptosis in ApoE-/- mice.

Abstract Funding Acknowledgements Type of funding sources: Public Institution(s). Main funding source(s): Fund for Scientific Research (FWO) Flanders INSPIRE project (H2020-MSCA-ITN program) Background The chemotherapeutic doxorubicin (DOX) is frequently used to treat a wide variety of cancers, but the cardiotoxic side effects limits its clinical use in some patients. Additionally, DOX contributes to vascular toxicity, which may be an early manifestation of DOX-associated toxicity. Therefore, there is an interest to evaluate vascular function in patients receiving DOX-based treatment regimens. Purpose We aimed to assess arterial stiffness and endothelial dysfunction as potential markers of vascular toxicity in both DOX-treated cancer patients and a murine model. Methods Female breast cancer patients with a need for adjuvant or neoadjuvant DOX-based treatment were prospectively included. Chemotherapeutic regimen consisted of 12 weekly cycles of taxane treatment, followed by 4 cycles of DOX (and cyclophosphamide) treatment. Vascular function (endothelial function (FMD and RHI), arterial stiffness (Aix and cfPWV) and cardiac function (LVEF, hsTnI, NT-proBNP) were performed at baseline (T1), after completion of taxane treatment (T2) and after completion of DOX treatment (T3). In addition to the clinical study, male C57Bl6/J mice were intraperitoneally injected with 2 mg /kg (low dose) or 4 mg/kg DOX (high dose) once per week for 6 weeks. Arterial stiffness was assessed in vivo by measuring abdominal aorta pulse wave velocity (aaPWV) by high-frequency ultrasound imaging combined with ex vivo vascular function evaluation. Results Twenty patients (mean age 51.2 +/- 10.1 yrs) treated with DOX were included. After DOX treatment (T3 vs T2), left ventricular ejection fraction (LVEF) was decreased, while hsTnI and NT-proBNP levels were increased, indicating cardiotoxicity (Table 1). Likewise, LVEF was reduced in DOX-treated mice after 3 weeks, which persisted until the end of the treatment (6 weeks). In patients, RHI was impaired at T2 and T3 compared to T1, which indicates progressive endothelial dysfunction during treatment (Table 1). Consistent with these findings, DOX treatment in mice resulted in endothelial dysfunction, as evidenced by a lower basal nitric oxide (NO) index and reduced acetylcholine-induced endothelium-dependent vasorelaxation (Figure 1B & 1C). Finally, cfPWV was decreased at T2 and T3 compared to T1, even after correction for BP, suggesting rather an improvement of arterial stiffness (Table 1). Although DOX treatment in mice resulted in a similar trend towards lower aaPWV at the end of treatment (6 weeks), we observed an initial increase in aaPWV after 2 weeks (Figure 1A). Conclusion Collectively, these findings suggest that, apart from cardiotoxicity, DOX treatment results in early and consistent endothelial dysfunction, while evaluation of arterial stiffness may be time-sensitive. Hence, endothelial dysfunction may be a more reliable and sensitive marker than arterial stiffness to evaluate DOX-induced vascular toxicity in patients.

Maintenance of homeostasis in the upper small bowel is a vital process for the body and therefore highly controlled. The enteric nervous system and the endocrine system are the regulators in this process influencing each other. The endocrine system in the gut consists of the classical hormones [cholecystokinin (CCK) secretin] to evoke motility or secretion. They are under control of releasing factors which are probably influenced by the enteric nervous system. Diazepam binding inhibitor and luminal CCK-releasing factor are likely candidates for CCK-releasing peptides in the negative feedback process in the absence of pancreatic juice. Experimental evidence suggests a secretin-releasing peptide. Further studies will be needed to determine the physiological role of each of these peptides. Monitor peptide in the pancreatic juice seems to function as a specific positive enhancement for CCK release. All these peptides are inactivated by the proteolytic enzymes during the interdigestive period. The discovery of additional releasing peptides and factors is very likely.