Morphological comparative study of canine and feline diaphragms: Effect of bodyweight, body condition score, gender, age and breed.

The canine diaphragm is a muscular and tendinous dome-like plate and is largely involved in digestive and respiratory functions. Very few studies compared morphology of the diaphragm between dogs and cats and no studies have investigated the effects of individual factors on this morphology. So the aim of this study was to (1) study the effects of individual factors on the morphology of the diaphragm and (2) to compare its morphology between cats and dogs. Surface measurements of 86 diaphragms were performed. The tendinous centre (TC), the lumbar part of the peripheral muscular (LP) and the sternal and costal parts of the peripheral muscular (SCPM) were measured. Measurement ratios (surface of anatomical part of the diaphragm/total surface of the diaphragm) were used for statistical analysis (TC%S, LP%S, SCPM%S). The SCPM%S are significantly lower, and the LP%S are significantly higher when bodyweight increases in dogs and cats. The TC%S are significantly lower when the body condition score of dogs increases. The SCPM%S are significantly higher, and therefore, the TC%S and LP%S lower in brachycephalic animals as opposed to mesocephalic animals. The TC%S are significantly higher in dogs than in cats and the SCPM%S are significantly higher in cats than in dogs. This study proposed a cartography of the diaphragm morphology in cats and dogs taking into account individual animal factors. Significant differences in the diaphragm morphology between cats and dogs and between mesocephalic and brachycephalic animals were found. Further studies are necessary to confirm these results and to investigate the consequences of these variations.

Effectiveness and Safety of a Prolonged Hemodynamic Support by the IVAC2L System in Healthy and Cardiogenic Shock Pigs.

BACKGROUND: Mechanical circulatory supports are used in case of cardiogenic shock (CS) refractory to conventional therapy. Several devices can be employed, but are limited by their availability, benefit risk-ratio, and/or cost. AIMS: To investigate the feasibility, safety, and effectiveness of a long-term support by a new available device (IVAC2L) in pigs. METHODS: Experiments were carried out in male pigs, divided into healthy (n = 6) or ischemic CS (n = 4) groups for a median support time of 34 and 12 h, respectively. IVAC2L was implanted under fluoroscopic and TTE guidance under general anesthesia. CS was induced by surgical ligation of the left anterior descending artery. An ipsilateral lower limb reperfusion was created with the Solopath® system. Reperfusion was started after 1 h of support in healthy pigs and upon IVAC2L insertion in CS pigs. Hemodynamic and biological parameters were monitored before and during the whole period of support in each group. RESULTS: Occurrence of an ipsilateral lower limb ischemia was systematic in healthy and CS pigs in a few minutes after IVAC2L implantation, and could be reversed by the arterial reperfusion, as demonstrated by distal transcutaneous pressure in oxygen (TcPO2) and lactate normalization. IVAC2L support decreased pulmonary capillary wedge pressure (PCWP) (15.3 ± 0.3 vs. 7.5 ± 0.9 mmHg, p < 0.001), increased systolic blood pressure (SBP) (70 ± 4.5 vs. 101.3 ± 3.1 mmHg, p < 0.01), and cardiac output (CO) (4.0 ± 0.3 vs. 5.2 ± 0.6 l/min, p < 0.05) in CS pigs; at CS onset and after 12 h of support, without effects on heart rate or pulmonary artery pressure (PAP). Non-sustained ventricular arrhythmias were frequent at implantation (50%). A non-significant hemolysis was observed under support in CS pigs. Bleedings were frequent at the insertion and/or operating sites (30%). CONCLUSION: Long-term support by IVAC2L is feasible and associated with a significant hemodynamic improvement in a porcine model. These preclinical data open the door for a study of IVAC2L in human ischemic CS, keeping in mind the need for systematic reperfusion of the lower limb and the associated risk of bleeding.

Tolerance of torasemide in cats with congestive heart failure: a retrospective study on 21 cases (2016-2019).

BACKGROUND: In dogs with congestive heart failure (CHF), the efficacy of torasemide, a loop diuretic, has been demonstrated. However, unlike in dogs and humans little has been described about the use of torasemide in the cat with spontaneous CHF. The objectives of this retrospective study were therefore to describe the therapeutic use of oral torasemide in cats with spontaneous CHF, document its potential adverse effects while reporting the clinical course of this feline population following torasemide administration in addition to standard medical therapy. RESULTS: Medical records of 21 client-owned cats with CHF (median age = 10.6 years [interquartile range (IQR) = 6.5-11.2]) receiving torasemide were reviewed. Data collected included torasemide dosages, other concurrent medications, physical examination features, echocardiographic data, and potential adverse effects during follow-up. A survival analysis was performed to estimate the time from diagnosis to cardiac death. Dyspnea related to CHF was identified in all cats (pleural effusion [8/21], pulmonary edema [5/21] or both [8/21]), associated with ascites in 4/21 cats. The CHF cause was determined by echocardiography in all cats: hypertrophic (n = 10), restrictive (n = 6), arrhythmogenic right ventricular (n = 3), dilated (n = 1) cardiomyopathies, and aortic valve abnormality (n = 1). At initiation, median torasemide dosage was 0.21 mg/kg [IQR = 0.17-0.23] q24h. Clinical signs declined in most cats (20/21) during the first 2 weeks with no remarkable adverse events. Median survival time after torasemide prescription was 182 days [IQR = 46-330]. A contemporary control group including 54 cats with CHF, receiving furosemide as sole loop diuretic treatment was compared with the study group. Median (IQR) survival time of cats in the control group was not significatively different (p = 0.962) from that of the torasemide group, i.e., 148 days (9-364), although the torasemide group included significantly more cats with recurrent episodes of CHF (52%) that the control group (19%). CONCLUSIONS: This case series demonstrates that torasemide can be used in cats with spontaneous CHF. This therapeutic interest needs to be confirmed by prospective clinical trials.

Absence of SARS-CoV-2 infection in cats and dogs in close contact with a cluster of COVID-19 patients in a veterinary campus.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which originated in Wuhan, China, in 2019, is responsible for the COVID-19 pandemic. It is now accepted that the wild fauna, probably bats, constitute the initial reservoir of the virus, but little is known about the role pets can play in the spread of the disease in human communities, knowing the ability of SARS-CoV-2 to infect some domestic animals. In this cross-sectional study, we tested the antibody response in a cluster of 21 domestic pets (9 cats and 12 dogs) living in close contact with their owners (belonging to a veterinary community of 20 students) in which two students tested positive for COVID-19 and several others (n = 11/18) consecutively showed clinical signs (fever, cough, anosmia, etc.) compatible with COVID-19 infection. Although a few pets presented many clinical signs indicative for a coronavirus infection, no antibodies against SARS-CoV-2 were detectable in their blood one month after the index case was reported, using an immunoprecipitation assay. These original data can serve a better evaluation of the host range of SARS-CoV-2 in natural environment exposure conditions.

Influence of extracorporeal membrane oxygenation on the pharmacokinetics of ceftolozane/tazobactam: an ex vivo and in vivo study.

BACKGROUND: Extracorporeal membrane oxygenation (ECMO) is increasingly used in intensive care units and can modify drug pharmacokinetics and lead to under-exposure associated with treatment failure. Ceftolozane/tazobactam is an antibiotic combination used for complicated infections in critically ill patients. Launched in 2015, sparse data are available on the influence of ECMO on the pharmacokinetics of ceftolozane/tazobactam. The aim of the present study was to determine the influence of ECMO on the pharmacokinetics of ceftolozane-tazobactam. METHODS: An ex vivo model (closed-loop ECMO circuits primed with human whole blood) was used to study adsorption during 8-h inter-dose intervals over a 24-h period (for all three ceftolozane/tazobactam injections) with eight samples per inter-dose interval. Two different dosages of ceftolozane/tazobactam injection were studied and a control (whole blood spiked with ceftolozane/tazobactam in a glass tube) was performed. An in vivo porcine model was developed with a 1-h infusion of ceftolozane-tazobactam and concentration monitoring for 11 h. Pigs undergoing ECMO were compared with a control group. Pharmacokinetic analysis of in vivo data (non-compartmental analysis and non-linear mixed effects modelling) was performed to determine the influence of ECMO. RESULTS: With the ex vivo model, variations in concentration ranged from - 5.73 to 1.26% and from - 12.95 to - 2.89% respectively for ceftolozane (concentrations ranging from 20 to 180 mg/l) and tazobactam (concentrations ranging from 10 to 75 mg/l) after 8 h. In vivo pharmacokinetic exploration showed that ECMO induces a significant decrease of 37% for tazobactam clearance without significant modification in the pharmacokinetics of ceftolozane, probably due to a small cohort size. CONCLUSIONS: Considering that the influence of ECMO on the pharmacokinetics of ceftolozane/tazobactam is not clinically significant, normal ceftolozane and tazobactam dosing in critically ill patients should be effective for patients undergoing ECMO.

Appetite-stimulating effect of gabapentin vs mirtazapine in healthy cats post-ovariectomy.

OBJECTIVES: The aim of the study was to evaluate the appetite-stimulating effect of gabapentin by comparing it with mirtazapine in healthy cats in the first 8 h after ovariectomy surgery. METHODS: This double-masked, placebo-controlled, prospective clinical trial included 60 healthy cats presented to the hospital for ovariectomy: 20 received gabapentin, 21 received mirtazapine and 19 received a placebo immediately before and 6 h after surgery. Food was offered at 2, 4, 6 and 8 h post-ovariectomy. After each meal, food intake was measured. Data were analysed using repeated-measure ANOVA and a linear mixed-model analysis. Post-hoc Tukey's honest significant difference test was performed for multiple comparisons. RESULTS: Food intake increased in both treatment groups vs placebo. No statistically significant difference was found between cats treated with gabapentin or mirtazapine. CONCLUSIONS AND RELEVANCE: Cats receiving gabapentin ate more than cats in the placebo group. Thirty percent of cats in the gabapentin group covered their resting energy requirements, while none of the cats in the placebo group did. Gabapentin and mirtazapine produced similar effects on food intake.

Heated intravenous fluids alone fail to prevent hypothermia in cats under general anaesthesia.

Objectives The objective was to evaluate the clinical efficiacy of a constant rate infusion of heated fluid as the sole means of preventing intraoperative hypothermia in cats. Methods This randomised, prospective, clinical study was conducted at a university teaching veterinary hospital. Female cats (American Society of Anesthesiologists [ASA] grade I) undergoing elective surgery by laparotomy under general anaesthesia (acepromazine 0.05 mg/kg SC; morphine 0.2 mg/kg IV; propofol IV titrated, isoflurane 2% in 100% oxygen) were randomised in two groups. Both groups were infused with fluid (NaCl 0.9%, 5 ml/kg/h) either at room temperature (control group) or prewarmed at 43°C (warmed group) using an Astoflo Plus eco (Stihler Electronic) fluid heating device. No other heating device was used. Temperature, heart rate, respiratory rate and SpO2 were evaluated after induction (T0) and every 15 mins for 1 h (T15, T30, T45, T60). Mean arterial blood pressure was recorded every 30 mins (T0, T30 and T60). Results Thirty-four female cats (ASA grade I) were enrolled in the study. There was no difference in age, weight, propofol dose or room temperature (22.4 ± 1.1°C vs 22.0 ± 1.5°C; P = 0.363) between control and warmed groups, respectively. In both groups, oesophageal temperature significantly decreased during anaesthesia ( P <0.0001). The temperature decrease after 1 h was -3.6 ± 0.7°C in the warmed group and was not significantly different from the control group (-3.4 ± 0.7°C; P = 0.307). The slopes of the temperature decrease did not significantly differ between the two groups (-0.058 ± 0.013°C/min vs -0.060 ± 0.010°C/min for the control and warmed groups, respectively; P = 0.624). Conclusions and relevance This study provides clinical evidence that a constant rate infusion of heated fluid alone fails to prevent intraoperative hypothermia in cats. The low infusion rate (5 ml/kg/h) could partly explain the ineffectiveness of this active warming device in minimising or delaying the onset of intraoperative hypothermia.

Enoxaparin has no significant anticoagulation activity in healthy Beagles at a dose of 0.8 mg/kg four times daily.

Enoxaparin at 0.8 mg/kg every 6 h is used in dogs to prevent prothrombotic events, but has only been validated in healthy Greyhounds, a breed with documented hypocoagulable haemostatic profiles in vitro and bleeding tendencies in vivo. This study investigated the effects of enoxaparin at this dose rate on the coagulation parameters of eight healthy adult Beagles over a 48 h period. Anti-Xa activity was significantly increased 3 h after the second and third of nine injections, and target anti-Xa activity was only reached transiently in 3/8 dogs. Paradoxically, a transient increase in endogenous thrombin potential was observed 6 h after the third injection. Other haematologic, biochemical and coagulation parameters were not significantly altered from the baseline values during enoxaparin treatment (P > 0.05). This study suggests that assumptions regarding the pharmacodynamic effects of enoxaparin are not generalisable across breeds.

Intravenous lipid emulsion therapy in 20 cats accidentally overdosed with ivermectin.

OBJECTIVE: To describe the outcome of 20 cats treated with intravenous lipid emulsion (IVLE) after an accidental parenteral ivermectin overdose. CASE SERIES SUMMARY: Twenty adult cats presented after receiving a 4 mg/kg accidental subcutaneous overdose of ivermectin. After admission, two IVLE treatments were initiated in asymptomatic cats: a single bolus (1.5 mL/kg; n = 16) versus a bolus followed by a 30-minute constant rate infusion (0.25 mL/kg/min; n = 4). Six out of the 16 cats that received only the single bolus developed clinical signs of ivermectin intoxication. Based on the severity of the clinical signs and their duration (approximately 48 hours), these 6 cats were retrospectively considered either moderately (n = 3) or severely (n = 3) intoxicated by ivermectin. Cats with a low body condition score (BCS) had more severe signs of ivermectin toxicity. Additional IVLE was administered until clinical resolution was complete. Median (min to max) cumulative dose of IVLE per cat was 4.5 (3.0-4.5) mL/kg for 36 (12-36) hours and 19.5 (7.5-37.5) mL/kg for 96 (72-168) hours for moderately and severely intoxicated cats, respectively. NEW OR UNIQUE INFORMATION PROVIDED: Our series describes the treatment of accidental ivermectin parenteral overdose in 20 cats with early initiation of IVLE therapy. An early bolus followed by a 30-minute constant rate infusion of IVLE appeared to mitigate the signs of ivermectin toxicosis in cats compared to a single treatment bolus. Our observations also suggest that cats with a low BCS given only a bolus of IVLE treatment were more likely to develop signs of ivermectin intoxication and require a greater amount of IVLE for the resolution of clinical signs. Based on our observations, BCS appears to influence the severity of ivermectin toxicity with a low BCS being associated with more severe signs of ivermectin toxicity.

Anatomical and ultrasonographic study of the femoral nerve within the iliopsoas muscle in beagle dogs and cats.

OBJECTIVES: An ultrasound (US)-guided ventral suprainguinal approach to block the femoral nerve (FN) within the iliopsoas muscle (IPM) has recently been described in dogs. The goal of the present study was to provide the operator with additional information to locate the FN within the IPM in dogs and cats using US. STUDY DESIGN: The study was carried out in three phases: a dissection of the FN (phase 1); an in vivo US-assisted nerve study (phase 2), and an anatomical cross-sectional study (phase 3). ANIMALS: Nine healthy adult beagle dogs and nine healthy adult cats. METHODS: Dissections were performed to investigate the anatomical characteristics of the FN and its related structures in one dog and one cat. Ultrasound scans of the left and right FN were performed in eight dogs and eight cats. The FN diameter and the distance between the FN and the external iliac artery (EIA) in US images and in anatomical cryosections were measured. RESULTS: The median FN diameter did not differ significantly between cats and dogs (1.1 mm versus 1.0 mm) or between the two techniques (US versus anatomical cross-sectional study) (1.1 mm versus 1.1 mm in dogs; 1.0 mm versus 1.1 mm in cats). The US and anatomical measurements of the median distances between the FN and EIA differed significantly between dogs and cats (8.2 mm versus 5.8 mm by US; 5.7 mm versus 4.8 mm in the anatomical study). CONCLUSIONS AND CLINICAL RELEVANCE: The distance between the EIA and FN is reproducible in beagle dogs and cats and can be used in locating the FN within the IPM.

In vivo pharmacodynamic interactions between two drugs used in orthostatic hypotension--midodrine and dihydroergotamine.

A combination of midodrine and dihydroergotamine (DHE) is frequently used clinically in patients suffering from severe orthostatic hypotension (OH). Whereas midodrine acts as a selective, peripheral alpha1-receptor agonist, DHE displays complex pharmacology and can behave as an alpha-adrenergic receptor agonist or antagonist. Surprisingly, the consequences of such a combination on blood pressure have never been investigated. The present study was performed in order to evaluate the pressor effects induced by the administration of both midodrine and DHE in old conscious dogs (n = 6) in experimental condition reproducing autonomic failure-related baroreflex dysfunction (atropine 0.1 mg/kg). For this purpose, we first studied the relative potency and intrinsic activity of each agonist and noradrenaline (NA) for the alpha1-adrenergic receptor. The orders of potency obtained in our study were 0.35, 11 and 400 microg/kg for NA, DHE and midodrine, and intrinsic activity: NA > midodrine > DHE. These results strongly suggest that DHE really acts in vivo as an alpha1-adrenoceptor partial agonist. Afterwards, the pressor effects of coadministration of midodrine (0.4 mg/kg) and DHE (15 microg/kg) were investigated: in one setting, midodrine was first administered, followed by DHE; in another, DHE was first administered, followed by midodrine. Our results show that in conscious dogs, the combination of midodrine and DHE leads to near-complete abolition of the pressor effect induced by the first administered drug. This in vivo proof of such antagonistic effects on blood pressure could explain clinical observations of worsening of OH in humans administered midodrine plus DHE. Although in vivo results obtained in conscious healthy dogs need to be experimentally and clinically confirmed in humans suffering from OH, these results strongly suggest that a midodrine-DHE combined treatment should be avoided in clinical practice.

Kinetic analysis of cardiac transcriptome regulation during chronic high-fat diet in dogs.

In the present study, we investigated, using custom dog cDNA arrays, the time course of transcriptional changes in the left ventricle of dogs fed a normal diet or a high-fat diet (HFD) for 9-24 wk. Array hybridizations were performed with complex probes representing mRNAs expressed in left ventricles from obese hypertensive and lean control dogs. We identified 63 differentially expressed genes, and expression of 17 of 20 randomly chosen genes was confirmed by real-time PCR. Transcripts were categorized into groups involved in metabolism, cell signaling, tissue remodeling, ionic regulation, cell proliferation, and protein synthesis. Hierarchical clustering indicated that the pattern of coregulated genes depends on duration of the HFD, suggesting that HFD-induced obesity hypertension is associated with continuous cardiac transcriptome adaptation despite stability of both body weight and blood pressure. GenMAPP analysis of the data pointed out the crucial importance of the ventricle TGF-beta pathway. Our results suggest that this system may be involved in molecular remodeling during HFD and in changes observed in the transcription profile, reflecting functional and morphological abnormalities that arise during prolonged HFD. These results also suggest some novel regulatory pathways for cardiac adaptation to obesity.

Adrenomedullin upregulates M2-muscarinic receptors in cardiomyocytes from P19 cell line.

1. The effects of AM on expression of muscarinic (M) receptors from P19-derived cardiomyocytes were examined. 2. RT-PCR experiments revealed expression of M(1)-M(4) receptor genes. Immuno-histochemistry indicated that M(2) expression is restricted to contractile cells. Carbachol inhibition of isoprenaline-induced increase in beating rate was prevented by atropine and methoctramine (pA(2): 8.1). Inhibition of [(3)H]-NMS binding by atropine (pK(i): -8.4+/-0.2) and methoctramine (pK(i): -8.3+/-0.2) suggests that M(2) is the functional expressed isoform. 3. [(3)H]-NMS binding and semiquantitative RT-PCR studies showed a dome shaped time course of M(2) expression with a maximum at 7 days of differentiation followed by a progressive decline. 4. AM concentration-dependently upregulated M(2) receptor mRNA during late differentiation stages in P19 cells but also in rat atrial cardiomyocytes. This effect was potentiated by factor H. AM (100 nM) plus factor H (50 nM) treatment of P19 cells for 24 h significantly increased [(3)H]-NMS-specific binding (B(max): 81+/-7 vs 31+/-6 fmol mg(-1) prot). The effect of AM on mRNA levels was prevented by AM receptor antagonist AM(22-52) (1 micro M) but not by CGRP antagonist, CGRP(8-37) (1 micro M). 5. The mRNA levels encoding CRLR receptor declined with culture duration, whereas those encoding L1/G10D receptor remained stable. 6. Our findings demonstrate that AM regulates M(2) receptors expression in cardiomyocytes probably through a mechanism involving L1/G10D receptors. The 'in vivo' significance of this phenomenon remains to be demonstrated.

Endothelin-1 inhibits TNF alpha-induced iNOS expression in 3T3-F442A adipocytes.

1. Endothelin-1 (ET-1) and tumor necrosis factor alpha (TNFalpha) by their action on adipocytes have been independently linked to the pathogenesis of insulino-resistance. In isolated adipocytes, TNFalpha induces the expression of the inducible nitric oxide synthase (iNOS). The purpose of the present work was, in the 3T3-F442A adipocyte cell line, to characterise TNFalpha-induced iNOS expression and to determine whether or not ET-1 could influence TNFalpha-induced iNOS expression and NO production. 2. In differentiated 3T3-F442A, treatment with TNFalpha (20 ng ml(-1)) induced the expression of a functional iNOS as demonstrated by nitrite assay, Western blot, reverse transcription-polymerase chain reaction and Northern blot analysis. TNFalpha-induced iNOS expression requires nuclear factor kappaB activation, but does not necessitate the activation of the PI-3 kinase/Akt and P38-MAP kinase pathways. 3. ET-1, but not ET-3, inhibited the TNFalpha-induced expression of iNOS protein and mRNA as well as nitrite production. The effects of ET-1 were blocked by a specific ETA (BQ123, pA(2) 7.4) but not by a specific ETB receptor antagonist (BQ788). 3T3-F442A adipocytes express the mRNAs for prepro-ET-1 and the ET-A receptor subtype, but not for the ET-B subtype. 4. The inhibitory effect of ET-1 was not affected by bisindolylmaleimide, SB 203580 or indomethacin, inhibitors of protein kinase C, p38-MAP kinase and cyclooxygenase, respectively, and was not associated with cAMP production. However, the effect of ET-1 was partially reversed by wortmannin, suggesting the involvement of PI3 kinase in the transduction signal of ET-1. 5. Differentiated 3T3-F442A adipocytes did not release ET-1 with or without exposure to TNFalpha, although the mRNA for preproET-1 was detected in both pre- and differentiated adipocytes. 6. Thus, these results confirm that adipocytes are a target for circulating ET-1 and demonstrate that the activation of the ETA receptor subtype can prevent TNFalpha-induced iNOS expression.

Cardiac transcriptome analysis in obesity-related hypertension.

Obesity is associated with volumetric arterial hypertension and with early increase in heart rate and decreased heart rate variability. The consequences of obesity-related hypertension on heart gene regulation are poorly known and were investigated in a model of obesity-related hypertension induced by high fat diet in dogs. When compared with control animals (n=6), a 9-week high fat diet (n=6) provoked significant weight gain and increased blood pressure load and heart rate but failed to significantly change left ventricular mass assessed by echocardiography. Subtractive hybridization of dog heart cDNA libraries were used to generate sublibraries containing differentially expressed cDNAs that were in turn spotted onto membranes to create custom microarrays. Hybridizations of these microarrays with complex probes representing mRNAs expressed in right atria and left ventricles from obese hypertensive and control dogs were performed. Thirty-eight differentially expressed genes were identified; altered expression was confirmed by Northern blot analysis in 15. In addition, real-time quantitative polymerase chain reaction confirmed differential expression for 80% of the randomly chosen tested genes. Once identified, transcripts were categorized into groups involved in metabolism, cell signaling, ionic regulation, cell proliferation, protein synthesis, and tissue remodeling. In addition, we found a set of 11 cDNAs encoding proteins with unknown functions. This study clearly shows that obesity-related hypertension, lasting for only 9 weeks, causes marked changes in gene expression in right atrium as well as the left ventricle that may contribute to early functional changes in heart function and to long-term structural changes such as left ventricular hypertrophy and remodeling.

High isoproterenol doses are required to activate beta3-adrenoceptor-mediated functions in dogs.

The "in vivo" conditions for beta3-adrenoceptors (beta-AR) activation by isoproterenol were investigated in dog. Experiments were carried out in anesthetized dogs using isoproterenol as a nonselective beta-AR agonist. Intravenous infusion of isoproterenol (0.4 nmol/kg/min) induced arterial hypotension and tachycardia with a slight decrease in cutaneous blood flow. At this dose, isoproterenol increased glucose, glycerol, and nonesterified fatty acid plasma levels. The changes in cardiovascular and endocrine-metabolic parameters, induced by the low dose of isoproterenol, were suppressed by pretreatment with nadolol (1 mg/kg, i.v.). After nadolol administration, however, a 10-fold higher dose (4 nmol/kg/min) of isoproterenol was able to induce a decrease in arterial blood pressure with a slight tachycardia and an increase in cutaneous blood flow. This high dose of isoproterenol increased nonesterified fatty acid and glycerol plasma levels but failed to change glucose plasma levels. All these effects were abolished by a pretreatment with nadolol (1 mg/kg, i.v.) plus SR59230A [a selective beta3-adrenoceptor antagonist; (3-(2-ethylphenoxy)-1(1S)-1,2,3,4-tetrahydronaphth-1-ylaminol-(2S)2-propanol oxalate); 1 mg/kg, i.v.]. Moreover, as observed with the high dose of isoproterenol under nadolol pretreatment, an infusion of SR58611A [a selective beta3-adrenoceptor agonist; ((N2S)-7-carbethoxymethoxy-1,2,3,4-tetrahydronaphth-2-yl-(2R)-2-hydroxy-2-chlorophenyl) ethanamine hydrochloride] induces a decrease in mean arterial blood pressure associated with an increase in heart rate, cutaneous blood flow, and nonesterified fatty acid and glycerol plasma levels. These results demonstrate that the in vivo activation of beta3-adrenoceptors requires higher doses of catecholamine than those necessary for beta1- and/or beta2-adrenoceptor stimulation. These results also argue for the lack of a beta3-AR involvement in the control of heart rate and glycogenolysis in dogs.

Alpha2 -adrenoceptor function in arterial hypertension associated with obesity in dogs fed a high-fat diet.

OBJECTIVE: To investigate the status of alpha2-adrenoceptors in a model of obesity-related arterial hypertension. DESIGN: A parallel study in dogs randomly assigned to a high-fat diet (HFD group, n = 6) or normal canine food (controls, n = 6) for 9 weeks. METHODS: Postsynaptic vascular alpha2-adrenoceptors were assessed through analysis of dose-pressor responses to clonidine [2.5, 5.0 and 15.0 microg/kg intravenously (i.v.)] after muscarinic, beta- and alpha1-adrenergic receptor blockade. Presynaptic and central alpha2-adrenoceptors were studied through measurement of changes in plasma concentrations of catecholamine induced by yohimbine (0.05 mg/kg i.v.). The number of platelet alpha2-adrenoceptors (expressed as fmol/mg protein) and the percentage in a state of high affinity were measured using [3H]RX821002. RESULTS: Clonidine, when administered to dogs that were under autonomic blockade, elicited a dose-dependent increase in blood pressure. The doses of clonidine required to induce a 50% maximum increase in systolic and diastolic blood pressures remained unchanged after 9 weeks of a high-fat diet (systolic: 6.0 +/- 0.3 microg/kg at baseline and 5.6 +/- 0.2 microg/kg after 9 weeks; diastolic: 4.2 +/- 0.2 microg/kg at baseline and 3.9 +/- 0.2 microg/kg after 9 weeks). After 9 weeks of the regimen, plasma concentrations of noradrenaline were significantly greater in the HFD group than in controls (337 +/- 22 pg/ml compared with 212 +/- 37 pg/ml). The increment in plasma concentrations of noradrenaline elicited by yohimbine after 9 weeks was smaller in the HFD group than in controls (93 +/- 44% compared with 181 +/- 46%; P = 0.024). In the HFD group, the number of platelet alpha2-adrenoceptors and the percentage that were in a state of high affinity were significantly lower after 9 weeks, compared with baseline (number: 239 +/- 21 fmol/mg protein at baseline and 95 +/- 7 fmol/mg protein after 9 weeks; high-affinity: 30 +/- 3% at baseline and 21 +/- 4% after 9 weeks; P < 0.05). CONCLUSIONS: These results suggest that presynaptic or central alpha2-adrenoceptor function, or both, is specifically impaired after 9 weeks of a high-fat diet. These modifications may account for the development of arterial hypertension in this model.