Development of an in Vivo Lipopolysaccharide Inflammation Model to Study the Pharmacodynamics of COX-2 Inhibitors Celecoxib, Mavacoxib, and Meloxicam in Cockatiels (Nymphicus hollandicus).

Nonsteroidal anti-inflammatory drugs (NSAIDs) are used frequently in avian medicine for their antipyretic, analgesic, and anti-inflammatory properties during surgery and for diseases that cause tissue damage and inflammation. NSAIDs inhibit cyclooxygenase (COX) enzymes, which are responsible for the induction of pyresis, pain, and inflammation. In our study, a lipopolysaccharide-induced (LPS) pyresis model was optimized using cockatiels (Nymphicus hollandicus) as subject birds (four males/three females) and validated in two females and one male, characterized by an intravenous bolus injection of LPS (7.5 mg/kg) administered at T0 and T24 (24 hours following the first LPS injection). To demonstrate the feasibility of the model to assess pharmacodynamic (PD) parameters of different NSAIDs, mavacoxib 4 mg/kg (four males/four females), celecoxib 10 mg/kg (four males/four females) and meloxicam 1 mg/kg (four males/four females) were evaluated in the model at dosages used frequently in practice. The PD parameters (body temperature, mentation, posture, preference of location in the cage, and prostaglandin E2 [PGE2] plasma concentrations) were determined for 10 hours following the second LPS injection. At the doses evaluated, mavacoxib and celecoxib significantly reduced LPS-induced hypothermia, but had no clear effects on other clinical signs of illness. In contrast, no effect on hypothermia or clinical appearance was observed in the LPS-challenged cockatiels treated with meloxicam. All three NSAIDs were able to inhibit the increase in LPS-induced PGE2 plasma concentrations, yet the effect was most pronounced in the birds treated with meloxicam. Consequently, the presented model opens perspectives for future dose-effect PD studies to optimize analgesic protocols in cockatiels.

Biomechanical properties of celiotomy wounds closed with tape and cyanoacrylate versus intradermal sutures.

OBJECTIVE: To compare cutaneous wound closure with mesh and 2-octyl cyanoacrylate (MOC) vs an intradermal suture pattern (ISP) in terms of time of application and biomechanical properties. SAMPLE POPULATION: Experimental study with 12 female beagle dogs. METHODS: A skin incision was created from the xiphoid to the pubis as part of an ovariohysterectomy; the linea alba and subcutaneous tissue were closed routinely. Half of the skin incision was closed with MOC, and the other half was closed with an ISP. Tissue samples were collected from both sections at days 14 and 28 and tested for ultimate strength and stiffness. RESULTS: Closure with MOC (72.8 ± 14.0 s) was faster than with an ISP (398.4 ± 36.4 s; P = .001). The ultimate load and stiffness increased with time for MOC (P = .005 and P = .005, respectively) and ISP (P < .001 and P < .001, respectively). On day 14, ultimate load and stiffness were greater in wounds closed with MOC compared with ISP (P = .014 and P = .02, respectively). No difference between groups was detected at day 28. CONCLUSION: Cutaneous wound closure with MOC was faster and resulted in superior strength at 14 days compared with closure with an ISP in this healthy population. CLINICAL SIGNIFICANCE: Mesh and 2-octyl cyanoacrylate offers an attractive alternative to ISP for skin closure after celiotomy in dogs, especially if surgical/anesthesia time is a concern.

The Ontogeny of Cytochrome P450 Enzyme Activity and Protein Abundance in Conventional Pigs in Support of Preclinical Pediatric Drug Research.

Since the implementation of several legislations to improve pediatric drug research, more pediatric clinical trials are being performed. In order to optimize these pediatric trials, adequate preclinical data are necessary, which are usually obtained by juvenile animal models. The growing piglet has been increasingly suggested as a potential animal model due to a high degree of anatomical and physiological similarities with humans. However, physiological data in pigs on the ontogeny of major organs involved in absorption, distribution, metabolism, and excretion of drugs are largely lacking. The aim of this study was to unravel the ontogeny of porcine hepatic drug metabolizing cytochrome P450 enzyme (CYP450) activities as well as protein abundances. Liver microsomes from 16 conventional pigs (8 males and 8 females) per age group: 2 days, 4 weeks, 8 weeks, and 6-7 months were prepared. Activity measurements were performed with substrates of major human CYP450 enzymes: midazolam (CYP3A), tolbutamide (CYP2C), and chlorzoxazone (CYP2E). Next, the hepatic scaling factor, microsomal protein per gram liver (MPPGL), was determined to correct for enzyme losses during the fractionation process. Finally, protein abundance was determined using proteomics and correlated with enzyme activity. No significant sex differences within each age category were observed in enzyme activity or MPPGL. The biotransformation rate of all three substrates increased with age, comparable with human maturation of CYP450 enzymes. The MPPGL decreased from birth till 8 weeks of age followed by an increase till 6-7 months of age. Significant sex differences in protein abundance were observed for CYP1A2, CYP2A19, CYP3A22, CYP4V2, CYP2C36, CYP2E_1, and CYP2E_2. Midazolam and tolbutamide are considered good substrates to evaluate porcine CYP3A/2C enzymes, respectively. However, chlorzoxazone is not advised to evaluate porcine CYP2E enzyme activity. The increase in biotransformation rate with age can be attributed to an increase in absolute amount of CYP450 proteins. Finally, developmental changes were observed regarding the involvement of specific CYP450 enzymes in the biotransformation of the different substrates.

Postnatal Maturation of the Glomerular Filtration Rate in Conventional Growing Piglets As Potential Juvenile Animal Model for Preclinical Pharmaceutical Research.

Adequate animal models are required to study the preclinical pharmacokinetics (PK), pharmacodynamics (PD) and safety of drugs in the pediatric subpopulation. Over the years, pigs were presented as a potential animal model, since they display a high degree of anatomical and physiological similarities with humans. To assess the suitability of piglets as a preclinical animal model for children, the ontogeny and maturation processes of several organ systems have to be unraveled and compared between both species. The kidneys play a pivotal role in the PK and PD of various drugs, therefore, the glomerular filtration rate (GFR) measured as clearance of endogenous creatinine (Jaffe and enzymatic assay) and exo-iohexol was determined in conventional piglets aging 8 days (n = 16), 4 weeks (n = 8) and 7 weeks (n = 16). The GFR data were normalized to bodyweight (BW), body surface area (BSA) and kidney weight (KW). Normalization to BSA and KW showed an increase in GFR from 46.57 to 100.92 mL/min/m2 and 0.49 to 1.51 mL/min/g KW from 8 days to 7 weeks of age, respectively. Normalization to BW showed a less pronounced increase from 3.55 to 4.31 mL/min/kg. The postnatal development of the GFR was comparable with humans, rendering the piglet a convenient juvenile animal model for studying the PK, PD and safety of drugs in the pediatric subpopulation. Moreover, to facilitate the assessment of the GFR in growing piglets in subsequent studies, a formula was elaborated to estimate the GFR based on plasma creatinine and BW, namely eGFR =1.879 × BW1.092[Formula: see text].

Immunomodulatory properties of gamithromycin and ketoprofen in lipopolysaccharide-challenged calves with emphasis on the acute-phase response.

Macrolide antibiotics and non-steroidal anti-inflammatory drugs (NSAIDs) have been reported to be modulators of the innate immune response, irrespectively of their antimicrobial and anti-inflammatory actions. Therefore, it was our objective to evaluate whether the macrolide gamithromycin (GAM) and the NSAID ketoprofen (KETO) attenuate the acute-phase response in calves, and whether their combined administration is beneficial due to synergistic and/or additive effects. To this end, both drugs, as well as their combination, were studied in a previously developed inflammation model, i.e., the induction of an acute-phase response by an intravenous lipopolysaccharide (LPS) challenge (0.5 µg/kg body weight). Sixteen 4-week-old Holstein-Friesian calves were randomized into 4 groups: a positive control (+CONTR) group, receiving LPS but no pharmacological treatment (n=4) and a GAM (n=4), a KETO (n=4) and a GAM-KETO (n=4) group, receiving the respective drugs 1h prior to LPS administration. Clinical scoring and blood collection were performed at regular time points until 72 h post LPS challenge. Plasma concentrations of the selected cytokines (tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6)), acute-phase protein (serum amyloid A (SAA)) and prostaglandin E2 (PGE2) were subsequently quantified. Pre-treatment with GAM had no effect in the inflammation model compared to the +CONTR group. KETO, on the other hand, completely inhibited depression, anorexia and fever. This remarkable influence was associated with a significant reduction of PGE2 synthesis by KETO, while the effect on TNF-α, IL-6 and SAA was not straightforward. The combined administration of GAM and KETO provided no synergistic or additive effects in this model, neither clinically nor regarding the studied inflammatory mediators. In conclusion, KETO entirely inhibited PGE2 synthesis, fever development and depression, while GAM did not exert any effect in this model. These results promote the concomitant use of an antimicrobial drug and a NSAID in the treatment of calf diseases associated with LPS, both to enhance clinical recovery and to improve animal welfare.

Serologic screening for 13 infectious agents in roe deer (Capreolus capreolus) in Flanders.

INTRODUCTION: In order to investigate the role of roe deer in the maintenance and transmission of infectious animal and human diseases in Flanders, we conducted a serologic screening in 12 hunting areas. MATERIALS AND METHODS: Roe deer sera collected between 2008 and 2013 (n=190) were examined for antibodies against 13 infectious agents, using indirect enzyme-linked immunosorbent assay, virus neutralisation, immunofluorescence, or microagglutination test, depending on the agent. RESULTS AND DISCUSSION: High numbers of seropositives were found for Anaplasma phagocytophilum (45.8%), Toxoplasma gondii (43.2%) and Schmallenberg virus (27.9%), the latter with a distinct temporal distribution pattern following the outbreak in domestic ruminants. Lower antibody prevalence was found for Chlamydia abortus (6.7%), tick-borne encephalitis virus (5.1%), Neospora caninum (4.8%), and Mycobacterium avium subsp paratuberculosis (4.1%). The lowest prevalences were found for Leptospira (1.7%), bovine viral diarrhoea virus 1 (1.3%), and Coxiella burnetii (1.2%). No antibodies were found against Brucella sp., bovine herpesvirus 1, and bluetongue virus. A significant difference in seroprevalence between ages (higher in adults >1 year) was found for N. caninum. Four doubtful reacting sera accounted for a significant difference in seroprevalence between sexes for C. abortus (higher in females). CONCLUSIONS: Despite the more intensive landscape use in Flanders, the results are consistent with other European studies. Apart from maintaining C. abortus and MAP, roe deer do not seem to play an important role in the epidemiology of the examined zoonotic and domestic animal pathogens. Nevertheless, their meaning as sentinels should not be neglected in the absence of other wild cervid species.

Evaluation of heart rate variability in dogs during standard and microburst vagus nerve stimulation: a pilot study.

Vagus nerve stimulation (VNS) is an established treatment for epilepsy and depression in human patients, but in both humans and dogs, optimal stimulation parameters remain unknown. Delivering afferent bursts of stimulation may be promising as a means of increasing efficacy, but evaluation of potential effects on the heart due to unavoidable efferent stimulation is required. The present study investigated heart rate variability (HRV) in healthy Beagle dogs treated with 1 h of sham, standard or microburst left-sided VNS in a crossover design. No significant differences were found between the stimulation paradigms for any of the cardiac parameters. Short-term left-sided VNS, including a novel bursting pattern (microburst VNS), had no statistically significant effect on HRV in ambulatory healthy dogs. Studies in a larger number of animals with long-term VNS are recommended.

Long-term follow-up of atrial function after cardioversion of atrial fibrillation in horses.

Atrial fibrillation (AF) causes atrial electrical and contractile remodelling in horses. The aim of this study was to quantify left atrial (LA) contractile function and its time course of recovery after cardioversion of naturally-occurring AF in horses. The study population included 42 AF horses which were successfully treated using transvenous electrical cardioversion TVEC (n=39) or quinidine sulfate (n=3), with trivial or mild mitral regurgitation present in 25 horses. Thirty-seven healthy horses were used as controls. AF duration was estimated based on the history and previous examinations. Echocardiography was performed during general anaesthesia after TVEC (day 0) and on days 1, 2, 6 and then 7 weeks after cardioversion. The two-dimensional (2D) echocardiographic measurements included LA diameter, area and ejection phase indices such as fractional shortening. Atrial TDI measurements included peak myocardial velocity during atrial contraction (A), time to onset A, time to peak A and duration of A. During follow-up after cardioversion, atrial contractile function measured by 2D echocardiography and TDI gradually improved. At 7 weeks following cardioversion, TDI-based myocardial velocities returned to reference values. However, AF horses still showed significantly larger atrial dimensions, lower 2D ejection phase indices and prolonged TDI-based conduction time compared to the control group. In conclusion, AF-induced atrial contractile dysfunction gradually improves in the weeks following cardioversion, but at 7 weeks post-cardioversion, significant differences remain compared to healthy controls.

Exposure to the proton scavenger glycine under alkaline conditions induces Escherichia coli viability loss.

Our previous work described a clear loss of Escherichia coli (E. coli) membrane integrity after incubation with glycine or its N-methylated derivatives N-methylglycine (sarcosine) and N,N-dimethylglycine (DMG), but not N,N,N-trimethylglycine (betaine), under alkaline stress conditions. The current study offers a thorough viability analysis, based on a combination of real-time physiological techniques, of E. coli exposed to glycine and its N-methylated derivatives at alkaline pH. Flow cytometry was applied to assess various physiological parameters such as membrane permeability, esterase activity, respiratory activity and membrane potential. ATP and inorganic phosphate concentrations were also determined. Membrane damage was confirmed through the measurement of nucleic acid leakage. Results further showed no loss of esterase or respiratory activity, while an instant and significant decrease in the ATP concentration occurred upon exposure to either glycine, sarcosine or DMG, but not betaine. There was a clear membrane hyperpolarization as well as a significant increase in cellular inorganic phosphate concentration. Based on these results, we suggest that the inability to sustain an adequate level of ATP combined with a decrease in membrane functionality leads to the loss of bacterial viability when exposed to the proton scavengers glycine, sarcosine and DMG at alkaline pH.

Evaluation of tissue Doppler imaging for regional quantification of radial left ventricular wall motion in healthy horses.

OBJECTIVE: To compare the feasibility and repeatability of tissue Doppler imaging (TDI) for quantification of radial left ventricular (LV) velocity and deformation from different imaging planes and to correlate cardiac event timing data obtained by TDI to M-mode and pulsed-wave Doppler-derived time intervals in horses. ANIMALS: 10 healthy adult horses. PROCEDURES: Repeated echocardiography was performed by 2 observers from right and left parasternal short-axis views at papillary muscle and chordal levels. The TDI measurements of systolic and diastolic velocity, strain rate, strain peak values, and timing were performed in 8 LV wall segments (LV free wall and interventricular septum from right parasternal views; left and right region of LV wall from left parasternal views). The inter- and intraobserver within- and between-day variability and measurement variability were assessed. The correlation between TDI-based measurements and M-mode and pulsed-wave Doppler-based time measurements was calculated. RESULTS: TDI measurements of velocity, strain rate, and strain were feasible in each horse, although deformation could often not be measured in the LV free wall. Systolic and diastolic time intervals could be determined with low to moderate variability, whereas peak amplitude variability ranged from low to high. The TDI-based time measurements were significantly correlated to M-mode and pulsed-wave Doppler measurements. CONCLUSIONS AND CLINICAL RELEVANCE: TDI measurements of radial LV velocity and deformation were feasible with low to moderate variability in 8 LV segments. These measurements can be used for evaluating LV function in further clinical studies.

Influence of atrioventricular interaction on mitral valve closure and left ventricular isovolumic contraction measured by tissue Doppler imaging.

BACKGROUND: The influence of atrioventricular (AV) interaction on mitral valve closure (MVC) and left ventricular (LV) isovolumic contraction is not fully clarified. We investigated the relationship among AV delay, MVC, and LV isovolumic contraction using a horse model because of the low heart rate and physiologically long AV delay. METHODS AND RESULTS: Six horses were evaluated during sinus rhythm, right ventricular pacing without preceding atrial contraction, and dual-chamber pacing at AV delays of 150 to 350 ms, programmed at a constant rate. Right parasternal 4-chamber views were recorded for simultaneous measurements of MVC from anatomic M-mode and radial tissue Doppler-based LV pre-ejection velocity and isovolumic acceleration. During sinus rhythm and long AV delays (≥300 ms), 2 positive pre-ejection velocity peaks were present. The first peak was identified as LV recoil during atrial relaxation and consistently preceded MVC by 33±17 ms. The second peak was related to LV isovolumic contraction, occurring after MVC. This suggests that MVC was caused by atrial relaxation and followed by true isovolumic contraction. During short AV delays (<300 ms) and right ventricular pacing, MVC occurred significantly later. Only 1 pre-ejection peak was present, of which the end coincided with MVC with a mean difference of -1.5±10 ms. This suggests that LV contraction caused MVC. Peak velocity and isovolumic acceleration were significantly higher (P<0.001) because the mitral valve was open at the onset of LV contraction. CONCLUSIONS: Depending on the AV delay, MVC can be atrio- or ventriculogenic, resulting in significant alterations of the LV peak pre-ejection velocity and isovolumic acceleration.

The influence of morphine on cerebral 5-HT2A availability in dogs: a SPECT study.

UNLABELLED: The opioid and serotonergic systems are closely involved in pain processing and mood disorders. The aim of this study was to assess the influence of systemic morphine on cerebral serotonin 2A receptor (5-HT(2A)) binding in dogs using SPECT with the 5-HT(2A) radioligand (123)I-5I-R91150. METHODS: 5-HT(2A) binding was estimated with and without morphine pretreatment in 8 dogs. The 5-HT(2A) binding indices in the frontal, parietal, temporal, and occipital cortex and in the subcortical region were obtained by semiquantification. RESULTS: A significantly decreased 5-HT(2A) binding index was found in the morphine group for the right (morphine, 1.41 ± 0.06; control, 1.52 ± 0.10) and left (morphine, 1.44 ± 0.08; control, 1.55 ± 0.11) frontal cortices, with P = 0.012 and P = 0.040, respectively. No significant differences were noted for the other regions. CONCLUSION: Morphine decreased the frontocortical 5-HT(2A) availability, confirming an interaction between the 5-HTergic and the opioid systems. Whether this interaction is caused by decreased receptor density due to direct internalization or is the result of indirect actions, such as increased endogenous serotonin release, remains to be elucidated.

Viability and cell death of synovial fluid neutrophils as diagnostic biomarkers in equine infectious joint disease: a pilot study.

Synovial fluid samples from culture-confirmed infected joints (n=13), joints with pronounced non-infectious synovitis (n=11) and healthy joints (n=14) were collected from 24 equine patients and seven slaughterhouse horses. The samples from the joints with non-infectious synovitis and healthy joints served as negative controls. After isolation, counting and identification of neutrophils, the percentage viability, and the proportion apoptotic and necrotic neutrophils were determined by flow cytometry. Viability was significantly higher in infected samples compared to the controls. A significant difference in cell death type was observed, with apoptosis predominating in infected joints, and necrosis being more present in joints with pronounced non-infectious synovitis and healthy joints. The results of this pilot study suggest that flow cytometric analysis of neutrophil viability and cell death dynamics may assist the discrimination between infected and non-infected joints.

Flow cytometric detection of myeloperoxidase in horse neutrophils: a novel technique in equine diagnostic research.

Myeloperoxidase (MPO) is a protein of interest due to its involvement in equine pathologies. Until now, results in equine diagnostic research were achieved through extracellular MPO detection. However, studying the cellular MPO content in neutrophils has revealed important insights in human diseases. This study aimed to develop a technique for the specific detection of MPO on the single cell level defining a flow cytometric protocol for the detection of both equine surface-bound and cellular MPO. Both indirect and direct labeling techniques are described which include the comparison of two secondary antibodies and two linking-fluorochromes, respectively.

Stromal cell-derived factor-1 retention and cardioprotection for ischemic myocardium.

BACKGROUND: Stromal cell-derived factor-1 (SDF-1) is a chemoattractant of stem/progenitor cells, and several studies have shown that SDF-1 may improve ventricular function after infarction. SDF-1 is cleaved by proteases including matrix metalloproteinase-2 (MMP-2) and CD26/dipeptidylpeptidase-4 (DPP-4), which are activated in injured tissues. METHODS AND RESULTS: We investigated the biodistribution and functional roles of SDF-1 in experimental ischemia/reperfusion injury in rats. Radiolabeled SDF-1 given by intracoronary injection was selectively concentrated in ischemic myocardium. The enhanced uptake of SDF-1 in ischemic myocardium was not mediated by its receptor, CXCR4. Mass spectrometry and Western analyses showed that SDF-1 was cleaved by DPP-4 in plasma and myocardium, whereas a bioengineered MMP-2/DPP-4-resistant form of SDF-1, SSDF-1(S4V), was highly stable. A single dose of SSDF-1(S4V) exhibited greater potency for cardioprotection than wild-type SDF-1. SSDF-1(S4V) improved cardiac function in rats even after a 3-hour ischemic period. CONCLUSIONS: These results show that a single dose of protease-resistant SSDF-1(S4V) after myocardial infarction leads to dramatic improvement in angiogenesis and ventricular function even 3 hours after the onset of ischemia, revealing a simple, clinically feasible approach to prevention of heart failure.

Detection of innocent systolic murmurs by auscultation and their relation to hematologic and echocardiographic findings in clinically normal Whippets.

OBJECTIVE: To determine murmur prevalence by auscultation of 105 apparently healthy Whippets without signs of cardiac disease, to determine the origin of these murmurs, and to evaluate the influence of sex, type of pedigree (ie, bred for showing or racing), and training on these murmurs. DESIGN: Cross-sectional study. ANIMALS: 105 client-owned Whippets. PROCEDURES: All dogs were auscultated by the first author and underwent a complete physical and cardiological examination, together with a hematologic assessment. Several RBC variables and echocardiographic variables were compared between dogs with or without a murmur at the level of the aortic valve. RESULTS: 44 of 105 (41.9%) dogs had no murmur. A soft systolic murmur was present with point of maximal intensity at the level of the aortic valve in 50 (47.6%) dogs, at the level of the pulmonic valve in 8 (7.6%) dogs, and at the level of the mitral valve in 3 (2.9%) dogs. No significant differences were found in heart rate, rhythm, murmur presence, point of maximal intensity, and murmur grade between males and females, between dogs with race- and show-type pedigrees, or between dogs in training and not in training. Dogs with a murmur at the level of the aortic valve had a significantly higher aortic and pulmonic blood flow velocity and cardiac output, compared with dogs without a murmur. CONCLUSIONS AND CLINICAL RELEVANCE: Whippets have a high prevalence of soft systolic murmurs in the absence of any structural abnormalities, which fit the description of innocent murmurs. No influence of sex, pedigree type, or training was found on the occurrence of these murmurs in Whippets.

Electrocardiographic reference values in whippets.

The aim of this study was to determine the electrocardiographic characteristics of whippets and to compare the results with published reference values for a general dog population. Electrocardiographic parameters from 105 healthy whippets were used to establish reference values for the breed. The most important differences compared to published reference values were the higher median R-wave amplitudes in leads II, CV(6)LL and CV(6)LU. For some parameters (P-wave amplitude, ST-segment deflection and T-wave amplitude in lead II; R-wave amplitude in CV(5)RL), a marked percentage of the whippet values were above the published maximum reference data. The results confirmed that whippets have electrocardiographic characteristics similar to those reported in athletic heart syndrome in humans. Some of these characteristics could be erroneously taken as evidence of cardiac disease and clinicians should be aware of these factors to prevent unnecessary investigations in healthy dogs.

Enteral feeding reduces endothelial nitric oxide synthase in the caudal intestinal microvasculature of preterm piglets.

The initiation of enteral feeding represents a challenge to the neonatal intestinal microcirculation, especially in preterms where it predisposes to necrotizing enterocolitis (NEC). We hypothesized that a structural microvascular deficiency may occur when enteral feeding is initiated in preterm piglets susceptible to NEC. Stereologic volume densities of a pan-endothelial marker (vWF), and the main vasodilator endothelial nitric oxide synthase (eNOS), were determined along the small intestine of 1) unfed preterm piglets, 2) piglets receiving total parenteral nutrition (TPN) for 2-3 d, and 3) piglets fed 2 d sow's colostrum (TPN+SOW) or milk formula (TPN+FOR) following TPN. In the mucosa, vWF-density decreased in a cranio-caudal direction. A corresponding mucosal eNOS gradient appeared only after initiating enteral feeding. In TPN+SOW, eNOS induction may lag behind the mucosal growth of the caudal region. In TPN+FOR, formula-related factors (i.e. bacteria, cytokines) may suppress mucosal eNOS, indicated by increased stress-sensitive nuclear HIF1alpha staining. The low mucosal endothelial eNOS density was related to the presence of NEC lesions, maybe via increased hypoxia-sensitivity, especially in the caudal region as indicated by nuclear HIF1alpha-staining. Our results suggest an insufficient structural adaptation of the microvasculature to enteral feeding, especially of mucosal eNOS, which may lead to NEC.

Echocardiographic reference values in whippets.

The aim of the study was to establish reference echocardiographic values for whippets, to compare these values with previously published reference values for the general dog population, and to determine whether there is an influence of gender and breeding lines on echocardiographic measurements. Echocardiographic parameters from 105 apparently healthy whippets without cardiac symptoms were used to establish reference values for the breed and to compare these values with two previously reported reference ranges. The coefficients of the allometric equation Y= aM(b), useful to reconstruct normal M-mode and two-dimensional average values for whippets of varying weights, were calculated, as well as the lower and upper limits of the 95% prediction interval. First, we found that whippets have a significantly larger left ventricular diameter, increased left ventricular wall, and interventricular septum thickness than expected, in diastole as well as in systole. Fractional shortening was significantly lower than the reference value. Second, comparing males and females, taking body weight differences into account, females had a significantly larger left ventricular diameter in diastole and systole. Minor differences were found between racing and show pedigree dogs. In conclusion, the results of this study confirm that breed-specific reference values are needed in echocardiography. In whippets, the values found in this study can be used as references in order to avoid overinterpretation of cardiac dilation, hypertrophy, and/or decreased contractility in these dogs.

Vertebral heart size ranges specific for whippets.

To assess the influence of breed, breeding lines, and training on heart size, the vertebral heart size (VHS) was evaluated on left-to-right lateral, right-to-left lateral, dorsoventral, and ventrodorsal thoracic radiographs from 44 whippets free from cardiac and pulmonary disease. In lateral views, the VHS was 11.0 +/- 0.5 vertebrae (mean +/- SD) on right-to-left lateral and 11.3 +/- 0.5 vertebrae on left-to-right lateral radiographs, being larger than the 9.7 +/- 0.5 vertebrae proposed by Buchanan (P<0.0001). The VHS on left-to-right lateral views was larger than on right-to-left lateral views (P<0.0001). The VHS was 10.5 +/- 0.6 vertebrae on dorsoventral radiographs and 11.1 +/- 0.6 vertebrae on ventrodorsal radiographs. Both values were larger than the 10.2 +/- 1.5 vertebrae (dorsoventral) (P<0.0082) or 10.2 +/- 0.8 vertebrae (ventrodorsal) (P<0.0001) proposed by Buchanan. In addition, the VHS on ventrodorsal views was larger than on dorsoventral views (P<0.0001). Dogs out of racing pedigree lines had a significantly larger VHS than those out of show pedigree lines, and trained dogs had a significantly larger VHS than nontrained dogs. Because most trained dogs came out of racing pedigree lines, and all nontrained dogs came out of show pedigree lines, however, it is difficult to determine whether the higher VHS for trained dogs is due to genetic influences or training, or both. In conclusion, it is important to take into account the breed and the radiographic view when evaluating heart size in thoracic radiographs of whippets to avoid overinterpretation of cardiac enlargement in these dogs.