Variation of heat shock protein gene expression in the brain of cold-induced pulmonary hypertensive chickens

1. Quantitative real-time PCR was carried out to evaluate gene expression of heat shock proteins (HSP) (HSP27, HSP56, HSP60, HSP70, HSP90 and ubiquitin) in the brain (hindbrain, midbrain, forebrain) of chickens with cold-induced pulmonary hypertension.

2. The ratio of the right ventricle to the total ventricle (index of pulmonary hypertension in chickens) was increased in the cold-induced pulmonary hypertensive chickens at 42 d of age compared with control. The HSP genes were expressed in the three parts of the brain in the two experimental groups.

3. In the hindbrain of cold-induced pulmonary hypertensive chickens, the relative gene expression of HSP27, HSP60, HSP70 and HSP90 was decreased while gene expression of HSP56 and ubiquitin was increased compared with controls.

4. In the midbrain of cold induced-pulmonary hypertensive chickens, the expression of HSP56, HSP60, HSP70 and ubiquitin genes was increased compared with controls while HSP27 and HSP90 were decreased.

5. In the forebrain of cold induced-pulmonary hypertensive chickens, the expression of HSP56, HSP60, HSP70 and ubiquitin genes was increased while the expression of the HSP27 gene was decreased compared with controls.

6. It is concluded that overexpression of HSPs in the forebrain and midbrain probably delays the pathological process of cold stress whereas diminished expression of HSP genes in the hindbrain may affect the normal function of brain centres in this area to exacerbate pulmonary hypertension.

     

No expression of angiotensin II receptors and angiotensin-converting enzyme in myxomatous canine mitral valve leaflets. An autoradiographic study

The renin-angiotensin system, including angiotensin (Ang) II and angiotensin-converting enzyme (ACE), plays an important role in cardiac fibrous tissue formation. Since changes in valvular collagen are a central part of myxomatous mitral valve disease in the dog, we speculated that Ang II and ACE might play a role in the pathogenesis of this disease. In 10 mitral valves, five with and five without clear myxomatous changes, the presence and distribution of Ang II receptors and ACE was examined autoradiographically, using 125I-Ang II and 125I-lisinopril, respectively. At postmortem examination, diseased valves were taken from old dogs, control valves from young adult dogs. No significant level of Ang II and lisinopril binding was found in normal as well as diseased valve leaflets. Equally low, insignificant levels of 125I-Ang II binding were found in the myocardium of dogs with and without valvular disease. No significant level of myocardial 125I-lisinopril binding was found. The lack of autoradiographic evidence of Ang II receptors and ACE in normal and diseased canine mitral valve leaflets suggests that the renin-angiotensin system does not play a major role in the pathogenesis of the valvular changes.     

Comparative studies on the histochemical properties of M. iliotibialis lateralis from Kumamoto Cochin crossbred roaster and broiler chickens

1. Histochemical properties of M. iliotibialis lateralis were compared between Kumamoto Cochin (a Japanese native breed) crossbred roasters (KC roasters, 112 d of age) and normal broilers (56 d of age).

2. Myofibres were divided into Types II‐R, II‐I and II‐W showing high, moderate and low reduced nicotinamide adenine dinucleotide dehydrogenase (NADH‐DH) activities, respectively.

3. Subsarcolemmal formazan granules indicating very strong NADH‐DH activity in Type II‐R fibres were observed only in KC roasters. In both sexes the percentage of Type II‐R fibres present was greater in KC roasters than in broilers.

4. These results indicated marked differences in histochemical properties and meat quality between KC roasters and broilers.     

Angiotensin Converting Enzyme in Bovine Ovarian Follicular Fluid and its Relationship with Oestradiol and Progesterone

The purpose of the present study was to identify angiotensin converting enzyme (ACE) in bovine ovarian follicular fluid and to relate the ACE activity to the phase of the oestrous cycle, pregnancy, and the follicular fluid concentrations of oestradiol and progesterone. The ACE activity was similar to that found in bovine serum and was completely inhibited by the specific ACE inhibitor captopril. The 50% inhibitory concentration (IC₅₀) was 1.4 × 10^?8 mol/l (range 0.8 × 10^?8 to 5.0 × 10^?8 mol/l; n=6), which is similar to that found in bovine and human serum. The ACE activity did not differ in the pre‐ovulatory and luteal phase, pregnancy or cystic follicles. It correlated with the follicular fluid concentration of progesterone in cycling cows (ρ=0.476; p < 0.005; n=36), but did not correlate with the diameter of the follicles, the follicular fluid concentration of oestradiol or the ratio between the oestradiol and progesterone concentrations. The demonstration of ACE in bovine ovarian follicular fluid provides further evidence for the presence of a local renin–angiotensin system in the bovine ovary.     

血管紧张素转换酶2在维持肠道稳态中的作用

血管紧张素转换酶2(ACE2)是首先克隆出的人类血管紧张素转换酶(ACE)的同源基因,是肾素-血管紧张素系统(RAS)的关键调节因子,其主要通过水解血管紧张素Ⅱ(AngⅡ)生成血管紧张素1-7[Ang(1-7)]从而发挥舒张血管、抗炎、抗增殖等作用。近期研究发现,ACE2不仅对心血管和肾功能等具有明显的调控作用,对抑制肠道炎症、维持肠道稳态、保护肠道健康等同样具有积极的影响。当前对ACE2在肠道健康领域的研究较少。论文就ACE2通过影响肠道微生态对维持肠道稳态作用的研究进行综述,为发掘ACE2的新功能提供参考。     

Feeding reduced-protein diets to broilers subjected to hypobaric hypoxia is associated with the development of pulmonary hypertension syndrome

1. There are few studies on the effect of dietary protein content on pulmonary hypertension syndrome (PHS) in broiler chickens. Conflicting results from these studies prevent a clear conclusion on the effects of reduced-protein diets on development of the syndrome.

2. To obtain an understanding of the mechanisms involved, the current study, conducted at a high altitude (2100?m above sea level), evaluated the effect of three treatments that varied only according to dietary protein (CP) levels. One treatment with dietary CP advocated by National Research Council (1994) acted as a control. Two reduced-protein diets were also prepared with CP reduced 20 and 40?g/kg relative to the CP of the control, which were designated as LCPD2 and LCPD4, respectively. A total of 180?d-old male broilers (Ross 308) were randomised across 15 floor pens measuring 1.5?m² (12 birds per pen). Five such pens (replicates) were allotted to each dietary treatment. The protein treatments were applied from 1 to 42?d of age in which growth performance was measured and the mortality from PHS was monitored. At the end of trial (42?d), blood sampling was done and carcase characteristics were recorded.

3. Birds receiving LCPD4 gained more weight throughout the trial and had increased right ventricular weight ratio (RV:TV), relative liver weight, haematocrit, and heterophil: lymphocyte ratio at the end of the trial compared to the control. Plasma concentrations of nitric oxide (NO) and uric acid, however, were significantly lower in birds fed on LCPD4 than in those receiving the control diet.

4. Mortality from PHS was increased in birds fed the reduced-protein diets, which may have been associated with reduced concentration of plasma NO and increased haematocrit and RV:TV. In conclusion, feeding reduced-protein diets to broilers subjected to hypobaric hypoxia was associated with the development of PHS.     

Gene expression of endothelin-1 and its receptors in the heart of broiler chickens with T(3)-induced pulmonary hypertension

To investigate the effect of T₃-induced pulmonary hypertension on endothelin (ET) production and genes expression of ET-1, ET_A and ET_B receptors (ET_AR and ET_BR) during rearing, semiquantitative RT-PCR and enzyme immunometric assay were performed in the heart ventricles and serum, respectively. The ET-1 and its receptor genes were expressed in the right and left ventricles of control and T₃-treated broilers at 12, 28 and 49 days of age. There were significant (P < 0.05) reductions of the relative amounts of ET-1 (in both ventricles) and ET_AR (in the right ventricle) mRNAs at 28 and 49 days of age, in T₃-treated broilers compared to controls. The relative amounts of ET_BR mRNA in the right and left ventricles did not significantly differ between control and T₃-treated broilers at any age. The serum level of ET was significantly (P < 0.05) increased in T₃-treated chickens at 28 and 49 days of age when compared with that of the control. It is concluded that ET-1, ET_AR and ET_BR genes are normally expressed in the heart ventricles of broilers. It is likely that increased serum level of ET and decreased ET-1/ET_AR genes expression in the ventricles are involved in the heart dysfunction of broiler chickens with developmental pulmonary hypertension.     

Capturing the dynamics of systemic Renin‐Angiotensin‐Aldosterone System (RAAS) peptides heightens the understanding of the effect of benazepril in dogs

Mochel, J. P., Peyrou, M, Fink, M, Strehlau, G, Mohamed, R, Giraudel, J. M., Ploeger, B, Danhof, M. Capturing the dynamics of systemic Renin‐Angiotensin‐Aldosterone System (RAAS) peptides heightens the understanding of the effect of benazepril in dogs. J. vet. Pharmacol. Therap.  36 , 174–180. In dogs, activation of the Renin‐Angiotensin‐Aldosterone System (RAAS) is an important feature of congestive heart failure (CHF). Long‐term increases in angiotensin II (AII) and aldosterone (ALD) lead to the progression of heart failure to its end stage. Angiotensin‐converting enzyme inhibitors (ACEIs) are the foremost therapeutic option in the management of CHF. Recent literature has challenged the efficacy of ACEIs, based on modest reduction in urinary aldosterone (UALD) excretion despite marked inhibition of ACE activity. This study was designed to heighten the understanding of the effect of benazepril, a potent ACEI, on the RAAS, using a low‐sodium diet as an experimental model of RAAS activation. Time course profiles of RAAS peptides and related areas under the curve (AUC_{24 hours}) were used for comparison between benazepril and placebo groups. Results indicated substantial changes in the dynamics of these biomarkers. At presumed benazeprilat steady state, significant differences in AUC_{24 hours} of plasma renin activity (+90%), angiotensin I (+43%), and AII (?53%) were found between benazepril and placebo‐treated dogs. ALD decreased by 73% in plasma but only by 5% in urine. In conclusion, despite modest reduction in UALD excretion, benazepril markedly influences RAAS dynamics in dogs.     

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.     

Central histaminergic system interplay with suppressive effects of immune challenge on food intake in chicken

1. The aim of the current study was to investigate the interaction of the lipopolysaccharide (LPS) and histaminergic systems on appetite regulation in broilers. Effects of intracerebroventricular (ICV) injection of α-fluoromethylhistidine (α-FMH, histidine decarboxylase inhibitor), chlorpheniramine (histamine H₁ receptor antagonist), famotidine (histamine H₂ receptor antagonist) and thioperamide (histamine H₃ receptor antagonist) on LPS-induced hypophagia in broilers were studied.

2. A total of 128 broilers were randomly allocated into 4 experiments (4 groups and 8 replications in each experiment). A cannula was surgically implanted into the lateral ventricle. In Experiment 1, broilers were ICV injected with LPS (20 ng) prior to α-FMH (250 nmol). In Experiment 2, chickens were ICV injected with LPS followed by chlorpheniramine (300 nmol). In Experiment 3, broilers were ICV injected with famotidine (82 nmol) after LPS (20 ng). In Experiment 4, ICV injection of LPS was followed by thioperamide (300 nmol). Then, cumulative food intake was recorded until 4 h post-injection.

3. According to the results, LPS significantly decreased food intake. Chlorpheniramine significantly amplified food intake, and LPS-induced hypophagia was lessened by injection of chlorpheniramine. α-FMH, famotidine and thioperamide had no effect on LPS-induced hypophagia.

4. These results suggest that there is an interaction between central LPS and the histaminergic system where LPS-induced hypophagia is mediated by H₁ histamine receptors in 3 h food-deprived broilers.

     

Viral shedding and emission of airborne infectious bursal disease virus from a broiler room

1. The significance of airborne transmission in epidemics of infectious diseases in the livestock production industry remains unclear. The study therefore investigated the shedding route (faeces vs. exhaled air) of a vaccine strain of infectious bursal disease virus (IBDV) by broilers and the emission of airborne virus.

2. The experimental room contained 526 broilers which were orally inoculated at the age of 20?d. The airborne virus was sampled by three different bioaerosol samplers: Andersen six-stage impactor, all-glass impinger (AGI-30) and OMNI-3000.

3. Infected broilers started to shed virus in faeces on d 5 post inoculation (PI), and stopped shedding on d 12 PI. The faecal virus remained detectable for at least two d after drying under broiler room conditions. No virus was detected in the air exhaled by broilers.

4. Airborne virus was collected on d 5, 8 and 12 PI at 20?cm above the floor, and on d 8 and 12 PI in exhausted air. The emission rates of IBDV were 4·0 log₁₀ 50% tissue culture infectious dose (TCID₅₀)/bird/d on d 8 PI, and 4·5 log₁₀ TCID₅₀/bird/d on d 12 PI.

5. We concluded that broilers shed IBDV mainly through their faeces. The presence of indoor airborne virus is associated with the viral presence in faeces. The successful recovery of airborne virus in exhausted air indicates there is a potential risk of virus spreading to the ambient environment via air.     

Apocynin activity in spontaneously hypertensive rats (SHR): preliminary studies in vivo

An elevation in angiotensin II (Ang II) levels is a common occurrence in spontaneously hypertensive rats (SHRs). Infusions of Ang II and a high salt diet increase the activity of NADPH oxidase that stimulates superoxide anion (O⁻²) generation and increases the expression of certain subunits of NADPH oxidase. Apocynin, an NADPH oxidase inhibitor with antihypertensive effects, is able to inhibit the release of superoxide anion by inhibiting NADPH oxidase activity and blocking the migration of p47 phox to the mitochondrial membrane. The aim of our study was to evaluate the antihypertensive effects of apocynin in SHRs and Wistar rats (WKYs) using a micropuncture technique. After microperfusion of both the proximal and distal tubules, we found that SHRs treated with apocynin showed a decrease in the free-flow collection of the proximal tubule (PT), which was not affected in WKYs. Moreover, significant differences were not demonstrated in the distal tubule (DT), probably due a mechanism of compensation that occurs in the loop of Henle. In conclusion, it is possible that the mechanisms of reabsorption in the PT are controlled by the interactions of O⁻² and nitric oxide (NO). These data could suggest a higher activity of NADPH oxidase and increase in reactive oxygen species (ROS) production in the PT during hypertension.

     

Echocardiographic study of pulmonary hypertension syndrome in broiler chickens

Echocardiography was used to study cardiovascular structure and function during the development of pulmonary hypertension syndrome (PHS) in broiler chickens. Body weight-normalized right and left ventricular diameters at both end-diastole (RVDD, LVDD) and end-systole (RVDS, LVDS) were determined weekly in broilers reared under either normobaric (altitude, 96.7 m) or hypobaric conditions (simulated altitude, 2900 m) until 5 wk of age. Hypobaric-exposed broilers had larger RVDD at 3 and 4 wk of age and larger RVDS at 3, 4, and 5 wk of age. Hypobaric-exposed broilers also had larger LVDD at 2, 3, 4, and 5 wk of age and larger LVDS at 4 wk of age. Right (RVFS) and left ventricular fractional shortening (LVFS) were smaller in hypobaric- vs. normobaric-exposed broilers at 3, 4, and 5 wk of age and at 4 wk of age, respectively. Among hypobaric-exposed birds, PHS-positive (+) broilers had larger RVDD and RVDS than PHS-negative (-) broilers on week 3 and on weeks 1 and 3 after hypobaric exposure, respectively. PHS-positive (+) broilers also had smaller RVFS on week 1 after hypobaric exposure. Electrocardiographic and post-mortem data indicated that PHS+ broilers also developed right ventricular hypertrophy when compared with PHS-negative (-) broilers. These results are consistent with the hypothesis that PHS develops as a result of pulmonary hypertension and cardiac overload and suggest that PHS+ broilers have a greater and more persistent reaction to hypoxia than PHS- broilers.     

Interaction of transport distance and body weight on preslaughter stress and breast meat quality of broilers

1. The objective of the study was to investigate the effect of transport distance on blood metabolites and breast meat quality of broilers slaughtered at different weights.

2. The study was conducted on Ross 308 broilers from 27 different flocks reared under similar conditions. Slaughter weight was classified as <2·0?kg, 2·0–2·4?kg, and >2·4?kg. Transport distance was categorised as short (65?km), medium (115?km) and long (165?km) distance representing 90, 155 and 220?minutes at an average 45?km/h speed, for each slaughter weight.

3. Higher heterophils and heterophil:lymphocyte (H/L) ratios were obtained for broilers transported over a long distance. Long distance transport increased blood albumin, glucose, and triglycerides levels for <2·0?kg broilers, which did not differ from broilers slaughtered at >2·4?kg after long-distance transport.

4. Broilers slaughtered at >2·4?kg after long-distance transport had lower pH_u, and paler and tougher breast meat, than those broilers slaughtered at <2·0?kg after long-distance transport.

5. A negative correlation was obtained between pH_u and L*, thawing loss and texture. The L* value was negatively correlated with a*; and positively correlated with b*, thawing and cooking losses.

6. It was concluded that the effect of transport distance could not be evaluated independently of slaughter weight. The interaction between transport distance and slaughter weight contributes to preslaughter stress and meat quality.     

Effect of Angiotensin II Type 1 receptor blocker on cardiac angiotensin-converting enzyme and chymase-like activities, and cardiac fibrosis in cardiomyopathic hamsters

It has been reported that cardiac chymase has an effect on cardiac fibrosis through the Angiotensin (Ang) II formation and an Ang II-independent mechanism. In the present study, Ang II type 1 (AT1) receptor blocker (candesartan cilexetil) was administered to dilated cardiomyopathic (DCM; Bio TO2) hamsters for 4 weeks to study the effect of AT1 receptor blocker on cardiac chymase-like activity and cardiac fibrosis. Echocardiography, histological examination, and assessment of cardiac angiotensin-converting enzyme (ACE)/chymase-like activities were conducted. Hamsters showed cardiac dysfunction due to increased left ventricular dimensions and decreased ventricular wall thickness, significant increase in cardiac chymase-like activity, and fibrosis. This result indicates that the cardiac chymase-like activity is responsible for cardiac fibrosis. When candesartan cilexetil was administered to Bio TO2 hamsters, cardiac chymase-like activity increased significantly, whereas cardiac fibrosis decreased significantly. Cardiac ACE and chymase-like activities were unchanged in non-DCM hamsters with candesartan cilexetil. This suggests that the cardiac Ang II formation mechanism was stimulated by suppressing the effect of cardiac Ang II, and cardiac chymase-like activity could be increased. Moreover, this mechanism may be more highly activated if cardiac Ang II is activated in the heart. In conclusion, we demonstrated that AT1 receptor blocker reduced cardiac fibrosis, although cardiac chymase-like activity increased. Because the Ang II-forming pathway and the effect of chymase in hamsters is similar to that in dogs, the results of the present study may supplement the available information for dogs.     

Blood flow: a key regulatory component of corpus luteum function in the cow

Prostaglandin F2alpha (PGF2alpha) is the primary luteolysin in the cow. During the early luteal phase, the corpus luteum (CL) is resistant to the luteolytic effect of PGF2alpha. Once mature, the CL becomes responsive to PGF2alpha and undergoes luteal regression. These actions of PGF2alpha coincide with changes in luteal blood flow (BF): PGF2alpha has no effect on BF in the early CL, but acutely increases BF in the peripheral vasculature of the mature CL within 30 min of PGF2alpha injection. During spontaneous luteolysis, luteal BF increases on Days 17-18 of the estrous cycle, prior to any decrease in plasma progesterone (P). The increase in luteal BF is synchronous with an increase in plasma PGFM levels, suggesting that pulsatile release of PGF2alpha from uterus stimulates the increase in luteal BF. Serial biopsies of these CL showed that mRNA expression for endothelial nitric oxide synthase (eNOS) together with endothelin-1 (ET-1) and angiotensin converting enzyme (ACE) increases on Days 17-18 when the luteal BF is elevated. On Day 19 when plasma P level firstly decreases, eNOS mRNA returns to the basal level whereas ET-1 and ACE mRNA remains elevated. Cyclooxygenase-2 (COX-2) mRNA expression increases on Day 19. In support of these data, an in vivo microdialysis study revealed that luteal ET-1 and angiotensin II (Ang II) secretion increases and precedes PGF2alpha secretion during spontaneous luteolysis. In conclusion, we show for the first time that an acute increase of BF occurs in the peripheral vasculature of the mature CL together with increases in eNOS expression and ET-1 and Ang II secretion in the CL during the early stages of luteolysis in the cow. We propose that the increase in luteal BF may be induced by NO from large arterioles surrounding the CL, and simultaneously uterine or exogenous PGF2alpha directly increases ET-1 and Ang II secretion from endothelial cells of microcapillary vessels within the CL, thereby suppressing P secretion by luteal cells. Taken together, our results indicate that an acute increase in luteal BF occurs as a first step of luteolysis in response to PGF2alpha. Therefore, local BF plays a key role to initiate luteal regression in the cow.