N-carbamylglutamate restores nitric oxide synthesis and attenuates high altitudeinduced pulmonary hypertension in Holstein heifers ascended to high altitude

Background: High-altitude pulmonary hypertension(HAPH) is a life-threating condition for animals in high altitude,and disturbance of endothelial nitric oxide(NO) synthesis contributes to its pathogenesis. N-carbamylglutamate(NCG), which enhances arginine synthesis, promotes endogenous synthesis of NO. In this study, we determined the effects of NCG on alleviating HAPH in Holstein heifers that ascended to Tibet(Lhasa, 3,658 m).Methods: Exp. 1, 2,000 Holstein heifers were transported from low elevation(1,027 m) to Lhasa. After being exposed to hypoxia for 1 yr, Holstein heifers were assigned to a healthy group(Control, n = 6) with mean pulmonary hypertension(mPAP) 41 mmHg, and an HAPH affected group(HAPH, n = 6) with mPAP 49 mmHg.Lung tissues were collected to evaluate histopathological changes and the expression of endothelial nitric oxide synthase(eNOS). Exp. 2, ten healthy heifers and 10 HAPH affected heifers were supplemented with NCG(20 g/d per heifer) for 4 wk. Physiological parameters were determined and blood samples were collected on d-1 and d 28 of the feeding trial.Results: Expression of e NOS in small pulmonary arteriole intima was higher in the healthy than HAPH group(P = 0.006), whereas HAPH group had significantly thicker media and adventitia than healthy group(all P 0.05). The mRNA of eNOS and protein level of eNOS were higher in the lungs of heifers in the healthy group than in the HAPH group(both P 0.001), whereas endothelin-1 protein levels were higher in HAPH group than in the healthy group(P = 0.025). NCG supplementation decreased mPAP and ammonia(both P = 0.001), whereas it increased the expression of eNOS, arginine, and plasma NO(all P 0.05).Conclusions: The expression of eNOS was decreased in Holstein heifers with HAPH. NCG supplementation decreased m PAP through the restoration of eNOS and endogenous NO synthesis.     

Antioxidant supplementation of low‐protein diets reduced susceptibility to pulmonary hypertension in broiler chickens raised at high altitude

A reduced‐protein diet (designated as RPD) was prepared and its effects on growth performance and the development of pulmonary hypertension syndrome (PHS) were evaluated in broiler chickens compared to a normal‐protein diet (designated as NPD) or to the RPD supplemented with CoQ₁₀ alone (30 mg/kg) or in combination with vitamin E (30 mg/kg CoQ₁₀ + 100 mg/kg vitamin E). The RPD had 30 g/kg less crude protein compared to the NPD. A total of 208 1‐day‐old male broilers (Ross 308 strain) were used in a 42‐day trial. Serum concentrations of uric acid (UA) and nitric oxide (NO) significantly (p < 0.05) declined when chickens fed on the RPD. However, supplementing RPD with the antioxidants significantly (p < 0.05) increased the serum NO concentration. Although serum malondialdehyde (MDA) concentration was significantly (p < 0.05) higher in the RPD than the NDP, supplementing RPD with CoQ₁₀ and CoQ₁₀ + VE decreased serum MDA concentration to similar levels found in the NPD. Significant overexpression in GPX1 gene observed in the heart and lungs of broilers fed on the RPD, which was effectively restored by supplementation of CoQ₁₀. The right to total ventricular weight ratio (RV:TV) was significantly (p < 0.05) increased in birds fed the RPD, which concurred with an increase in mortality from pulmonary hypertension syndrome (PHS). However, a significant decline in mortality from PHS was observed when birds on RPD received CoQ₁₀ or CoQ₁₀ + VE. In conclusion, antioxidant supplementation effectively improves pulmonary hypertensive response in broiler chicken fed of reduced‐protein diets.     

Contribution of live heartworms harboring in pulmonary arteries to pulmonary hypertension in dogs with dirofilariasis

To investigate whether adult heartworms harboring in the pulmonary arteries contribute to pulmonary hypertension, we determined the cardio-pulmonary values immediately before and after removal of heartworms from the pulmonary arteries and before and after insertion of live worms in their place. In 10 heartworm-infected dogs, 8 to 46 worms were removed. The mean pulmonary arterial pressure fell significantly from 24.5 +/- 7.9 mmHg to 16.3 +/- 4.9 mmHg (p less than 0.01) immediately after removal. The right cardiac output decreased in 7 of the 10 cases. The total pulmonary resistance and right ventricular stroke work index also decreased. At 24 hours after removal, live heartworms were put back into the pulmonary arteries of their host dog. The mean pulmonary arterial pressure elevated significantly (p less than 0.01) immediately after insertion. The right cardiac output further decreased in 7 of the 10 dogs, and the total pulmonary resistance and right ventricular stroke work index increased. Separate from this, 12 to 42 heartworms were transplanted into the pulmonary arteries of 5 heartworm-free dogs. Immediately after transplantation, the pulmonary arterial pressure did not show any significant change. However, the stroke volume decreased, and the total pulmonary resistance increased. These facts suggest a contribution of live heartworms to the pulmonary hypertension, although there is a complicated interaction among the presence of heartworms, the pulmonary lesions and the pulmonary hypertension.     

Pulmonary arteriopathy and idiopathic pulmonary arterial hypertension in six dogs

Pulmonary arteriopathy (PA) is the pathologic hallmark in human medicine of diffuse constrictive (medial and intimal remodeling) or multifocal complex (plexiform and dilatative lesions) arterial lesions, or both, that lead to irreversible obliteration of the arterial lumen. Clinically, PA leads to pulmonary arterial hypertension (PAH), of which idiopathic (IPAH) is one of the 5 subsets, and ultimately, to right-sided heart failure (RHF). Clinical and pathologic findings from 6 dogs with diagnosis of IPAH and PA were reviewed. These dogs were of various pure (5/6, 83%) and mixed (1/6, 17%) breeding, 5 months to 9 years (mean 5.2 years) old, and predominantly female (4/6, 67%) and reproductively intact (4/6, 67%). Doppler echocardiography (n = 5) indicated increased pulmonary arterial pressures during systole (70-135 mm Hg, mean 98 mm Hg) and diastole (35-80 mm Hg, mean 58 mm Hg). All 6 dogs had right ventricular pressure overload, right ventricular eccentric hypertrophy, and RHF. Histologic examination confirmed the clinical diagnosis of IPAH in all dogs, revealing PA characterized by 1 of the 4 main human histologic subsets: 1) isolated medial hypertrophy (1/6, 17%); 2) medial hypertrophy-intimal thickening without the plexiform lesion (1/6, 17%); 3) medial hypertrophy-intimal thickening concurrent with the plexiform lesion, which often was regionally clustered and situated near branching points of the respiratory artery, the poststenotic dilatation lesion, and vasculitis (4/6, 66%); and 4) isolated arteritis (1/6, 17%). Ancillary lesions similar to those in humans also complicated the PA (5/6, 83%). The complex lesions and ancillary exudative alveolitis seemed to be important indicators of severe, likely rapidly progressive and fatal, IPAH.     

Sildenafil citrate therapy in 22 dogs with pulmonary hypertension

BACKGROUND: Pulmonary hypertension (PH) is a disease condition characterized by abnormally increased pulmonary artery pressures and often is associated with a poor prognosis. Sildenafil is a phosphodiesterase inhibitor that causes pulmonary arterial vasodilation and reduction in pulmonary artery pressures. HYPOTHESIS: Treatment with sildenafil will improve echocardiographic determinants of PH in dogs, while also improving quality of life and survival. ANIMALS: Twenty-two dogs with clinical and echocardiographic evidence of pulmonary hypertension. METHODS: A retrospective study evaluating the effects of sildenafil on physical examination, ECG and radiographic findings, blood pressure and echocardiographic findings of PH, clinical score, and outcome was completed. PH was defined as a peak tricuspid regurgitation flow velocity > or = 2.8 m/s or a peak pulmonic insufficiency flow velocity > or = 2.2 m/s. RESULTS: Sixteen of 22 dogs with PH were elderly females of small body size. Their clinical score was significantly improved (P = .0003) with sildenafil treatment, but physical examination findings remained unchanged. Heart rate, respiratory rate, vertebral heart size, ECG heart rate, and systolic blood pressure did not change significantly with sildenafil treatment (P > .05). Peak tricuspid regurgitation flow velocities did not change significantly with the treatment of sildenafil, but selected systolic time intervals were significantly improved. Survival times for all dogs ranged from 8 to > 734 days. CONCLUSIONS AND CLINICAL IMPORTANCE: Sildenafil did not significantly lower the degree of measurable PH in dogs. Clinical improvement and increased quality of life was seen with sildenafil treatment, despite lack of significant change in other variables.     

Caudal pulmonary artery to vein ratio on radiography can predict pulmonary hypertension in dogs with mitral regurgitation

Pulmonary hypertension (PH) is an important predictor of poor outcomes in dogs with mitral regurgitation (MR). The feasibility of radiography to predict PH in dogs with MR is unknown. This retrospective, observational, and analytic study aimed to identify a radiographic parameter to predict PH in dogs with MR. A total of 302 dogs diagnosed with MR on echocardiography were enrolled. Medical record and radiographic findings such as the size of the main pulmonary artery, left atrium, left ventricle, and right chamber, and cranial and caudal pulmonary arteries and veins were evaluated according to the presence of PH. The diameters of the cranial and caudal pulmonary vessels were compared to the fourth rib and the ninth rib, respectively, and the ratio of the pulmonary artery to the corresponding vein (CdPA/CdPV) was calculated. Pulmonary hypertension was diagnosed in 77 dogs (25.5%) and the prevalence of PH increased with MR grade. The CdPA/CdPV was significantly higher (P < 0.001) in the presence of PH. Multivariate analysis showed that the CdPA/CdPV was the only independent radiographic parameter that had a significant association with PH in dogs with MR (P = 0.028). The cut-off value of the CdPA/CdPV = 1.10 showed 90.6% specificity and 31.1% sensitivity for detecting PH in dogs with MR. In dogs with MR, PH can be predicted with high specificity when the caudal pulmonary artery is 1.1 times larger than the corresponding vein on radiographs.     

Haemodynamic effects of ATP in dogs during hypoxia-induced pulmonary hypertension

Pulmonary hypertension may result from an increase in vascular resistance caused by persistent hypoxia. We have investigated the effects of adenosine triphosphate (ATP), administered into the pulmonary artery, on haemodynamic changes occurring in anaesthetized adult dogs subjected to acute hypoxic pulmonary vasoconstriction. Hypoxia alone (ventilation with 10% O2/90% N2) caused significant increases in mean pulmonary arterial blood pressure (PAP), central venous pressure (CVP), and cardiac index (CI) by 71, 102 and 38%, respectively. ATP (0.03-3.0 micromol/kg/min approximately 0.02-1.65 mg/kg/min), when infused under hypoxic conditions, significantly reduced both mean PAP and systemic arterial blood pressure (ABP) in a dose-dependent manner. The maximum decrease in mean PAP amounted to 20%; mean ABP, on the other hand, was decreased by up to 52% (P<0.01). Heart rate, CI, CVP and pulmonary occlusion pressure were not dose-dependently affected by ATP. Our data indicate that while pulmonary arterial administration of ATP in mature dogs during hypoxic pulmonary hypertension causes dilation in the pulmonary vascular bed, it is even more effective in dilating the systemic vasculature. This result suggests a need for further evaluation and warrants cautious use of ATP in the treatment of hypoxic pulmonary hypertension in adult dogs.     

NT-proBNP, NT-proANP and cTnI concentrations in dogs with pre-capillary pulmonary hypertension

Objectives

To compare [NT-proBNP], [NT-proANP] and [cTnI] between control dogs with respiratory disease without pulmonary hypertension (PH) and dogs with pre-capillary PH, and to assess the accuracy of [NT-proBNP], [NT-proANP], [cTnI] to predict Doppler-derived peak tricuspid regurgitation (TR) gradient.

Animals

20 dogs. 8 control dogs with respiratory disease with no PH and 12 with pre-capillary PH.

Methods

[NT-proBNP], [NT-proANP] and [cTnI] were compared between the 2 groups and simple linear regression analysis was used to predict peak TR gradients from various blood biomarkers.

Results

Median [NT-proBNP] was higher in the dogs with PH (2011 pmol/L, 274–7713 pmol/L) compared to control dogs (744 pmol/L; 531–2710 pmol/L) (p = 0.0339). [NT-proBNP] was associated with peak TR gradient (R² = 0.7851, p = 0.0001). Median [NT-proANP] did not differ between dogs with PH (1747 fmol/L; 894–2884 fmol/L) and control dogs (1209 fmol/L; 976–1389 fmol/L (p = 0.058). [NT-proANP] was not associated with peak TR gradient (R² = 0.2780, p = 0.0781). Median [cTnI] did not differ between dogs with PH (0.2850 ng/mL; 0.19–1.13 ng/mL) and control dogs (0.2 ng/mL; 0.19–0.82 ng/mL, p = 0.3051). Median [TnI] was not associated with peak TR gradient (R² = 0.024, p = 0.6307).

Conclusions

[NT-proBNP] concentration is significantly higher in dogs with pre-capillary PH when compared to dogs with respiratory disease without PH, and [NT-proBNP] may be useful to predict the severity of estimated PH. Elevations in [NT-proBNP] due to pre-capillary PH may complicate the interpretation of [NT-proBNP] elevations in patients presenting with cardiorespiratory abnormalities. [NT-proANP] and [cTnI] were not elevated in dogs with pre-capillary PH.     

Evaluation of red cell distribution width in dogs with pulmonary hypertension

ObjectivesTo compare red cell distribution width (RDW) between dogs with different causes of pulmonary hypertension (PH) and a control dog population to determine whether RDW was correlated with severity of PH as measured by echocardiography. A further aim was to determine the prognostic significance of increased RDW for dogs with PH.AnimalsForty-four client-owned dogs with PH and 79 control dogs presented to a single tertiary referral institution.MethodsSignalment, clinical pathological and echocardiographic data were obtained retrospectively from the medical records of dogs with PH, and RDW measured on a Cell-Dyn 3500 was compared between dogs with pre- and post-capillary PH and a control population. Referring veterinary surgeons were contacted for follow-up information and Kaplan–Meier analysis was conducted to investigate differences in survival time between affected dogs with different RDW values.ResultsThe RDW was significantly greater in dogs with pre-capillary PH compared to control dogs. There was no difference in median survival times between dogs with PH divided according to RDW values. The RDW was positively correlated with mean corpuscular volume and haematocrit in dogs with PH, but did not correlate with echocardiographic variables.ConclusionsAn association was found between dogs with PH and increased RDW; however there was considerable overlap in values between control dogs and dogs with PH. The RDW was not associated with survival in this study.     

Comparison of a computed tomographic pulmonary trunk to aorta diameter ratio with echocardiographic indices of pulmonary hypertension in dogs

There are limited criteria for the detection of pulmonary hypertension in dogs undergoing computed tomography (CT) for pulmonary disease. This retrospective analytical exploratory study compared a CT pulmonary trunk to aorta ratio with echocardiographic estimates of pulmonary hypertension. Dogs having both a contrast thoracic CT and echocardiogram were selected and maximal pulmonary trunk and descending aorta diameters were measured by two observers on a single transverse CT image. Computed tomographic diameter ratios were compared with the echocardiographic parameters of tricuspid regurgitation gradient, right ventricular acceleration time‐to‐ejection time ratio, pulmonary insufficiency gradient, and pulmonary artery to aorta diameter. A total of 78 dogs were sampled, with 44 dogs having one or more finding suggestive of pulmonary hypertension. A moderate positive correlation was shown between tricuspid regurgitation gradient and CT pulmonary trunk to aorta ratio (r = 0.61, P‐value < 0.0001). Mean CT pulmonary trunk to aorta ratio of dogs with moderate (P = 0.0132) and severe (P < 0.0003) pulmonary hypertension were significantly higher than normal dogs. There was no significant difference in mean CT pulmonary trunk to aorta ratio between normal and mild pulmonary hypertension dogs (P = 0.4244). The intraclass correlation coefficient (0.72) showed good reproducibility of the ratio. Findings indicated that CT pulmonary trunk to aorta ratio is a reproducible and potentially useful method to predict moderate and severe pulmonary hypertension in dogs, but not mild pulmonary hypertension. In dogs undergoing thoracic CT for pulmonary disease, an increased ratio should prompt follow up echocardiography.     

The effect of dietary l-carnitine supplementation on pulmonary hypertension syndrome mortality in broilers exposed to low temperatures

Oxidative stress is involved in the development of pulmonary hypertension syndrome (PHS) in broilers. l-Carnitine has an antiperoxidative effect and supplemental l-carnitine has been revealed to increase broiler heart weight. The present study was conducted to evaluate the effect of an addition of 100 mg/kg l-carnitine to the basal diets on PHS mortality in cold-exposed broilers. Two-hundred and forty mixed-sex broilers were equally assigned to three groups. The control group was reared in normal temperatures throughout the experiment. Starting on day 14 continuing until the end of the experiment, the other two groups were subjected to a step-down temperature programme (by lowering the temperature 1-2 degrees C per day down to 12-14 degrees C) with or without l-carnitine added to the basal diets. Cold exposure increased the right/total ventricle ratio (RV/TV) and plasma malondialdehyde (MDA), reduced superoxide dismutase (SOD) and led to pulmonary vascular remodelling in birds without feeding additional l-carnitine. Supplemental l-carnitine reduced plasma MDA, increased SOD, inhibited remodelling and postponed the occurrence of PHS for 1 week in cold-exposed broilers; nevertheless, it did not significantly influence the cumulative PHS mortality (p > 0.05). On days 24 and 32, birds fed supplemental l-carnitine had lower RV/TV and higher total ventricle/body weight (p < 0.05) but unchanged right ventricle/body weight ratios (p > 0.05) compared to their cold-exposed counterparts, indicating an increase in left ventricle weight. However, from day 39 on, their RV/TV ratios were suddenly increased (p < 0.05). It was suggested that the l-carnitine-induced increase in left heart weight might partially account for the postponed occurrence of pulmonary hypertension in the early stage by elevating cardiac output, which might, in turn, lead to the resulting increase in pulmonary pressure. In view of its complex effects on cardiopulmonary haemodynamics, l-carnitine supplementation may be impractical for reducing PHS.     

Diagnostic value of echo-Doppler and tissue Doppler imaging in dogs with pulmonary arterial hypertension

BACKGROUND: Diagnosis of pulmonary arterial hypertension (PAH) relies on Doppler measurement of pulmonic and tricuspid regurgitation (TR). However, these are not always detectable. HYPOTHESIS: Tissue Doppler imaging (TDI), a novel noninvasive ultrasound technique, provides indirect but sensitive and specific assessment of elevated systolic pulmonary artery pressure (SPAP) in dogs. ANIMALS: One hundred and five dogs with TR. METHODS: Prospective observational study. Dogs were categorized as presenting normal (group 1, n = 45), mildly increased (group 2, n = 19), or moderately to severely increased (group 3, n = 41) SPAP, based on TR peak velocities (< 2.5, 2.5-3.0, and > 3.0 m/s, respectively). Ten quantitative echo-Doppler- and TDI-derived variables were assessed, including the main pulmonary arterial diameter to aortic diameter ratio, pulmonary flow acceleration time, and acceleration-to-ejection time ratio, the Tei index of right ventricular function, and 6 longitudinal basal right ventricular TDI variables. RESULTS: A significant correlation was observed between SPAP and each of the 10 tested variables (P < .05). Conventional echo-Doppler variables were less discriminating than the TDI for predicting increased SPAP. The combined systolic and diastolic right TDI index had the highest sensitivity and specificity (89% and 93% respectively, for a cutoff of 11.8 cm/s) and could discriminate between dogs in group 1 from dogs in group 2. CONCLUSIONS AND CLINICAL IMPORTANCE: TDI provided effective predictors of systolic PAH and demonstrated that both alterations in right-sided systolic and diastolic myocardial function can occur with mild increases in SPAP.