Effects of L-NAME, a non-specific nitric oxide synthase inhibitor, on AlCl3-induced toxicity in the rat forebrain cortex

The present experiments were done to determine the effectiveness of a non-specific nitric oxide synthase inhibitor, N-nitro-L-arginine methyl ester (L-NAME), on oxidative stress parameters induced by aluminium chloride (AlCl₃) intrahippocampal injections in Wistar rats. Animals were sacrificed 3 h and 30 d after treatments, heads were immediately frozen in liquid nitrogen and forebrain cortices were removed. Crude mitochondrial fraction preparations of forebrain cortices were used for the biochemical analyses: nitrite levels, superoxide production, malondialdehyde concentrations, superoxide dismutase (SOD) activities and reduced glutathione contents. AlCl₃ injection resulted in increased nitrite concentrations, superoxide anion production, malondialdehyde concentrations and reduced glutathione contents in the forebrain cortex, suggesting that AlCl₃ exposure promoted oxidative stress in this brain structure. The biochemical changes observed in neuronal tissues showed that aluminium acted as a pro-oxidant. However, the non-specific nitric oxide synthase (NOS) inhibitor, L-NAME, exerted anti-oxidant actions in AlCl₃-treated animals. These results revealed that NO-mediated neurotoxicity due to intrahippocampal AlCl₃ injection spread temporally and spatially to the forebrain cortex, and suggested a potentially neuroprotective effect for L-NAME.     

Effect of L-NAME on pulmonary arterial pressure,plasma nitric oxide and pulmonary hypertension syndrome morbidity in broilers

1. Experiments were conducted to evaluate the effect of a synthetic inhibitor of nitric oxide (NO) synthase (L-NAME) on pulmonary arterial pressure (PAP) and pulmonary hypertension syndrome (PHS) morbidity in broilers. 2. In Experiment 1, broilers were infused intravenously with L-NAME, and the mean pulmonary arterial pressure (mean PAP) and plasma NO were measured at 0, 1, 2 and 4 h after the start of infusion. The mean PAP increased and plasma NO was reduced at 1 to 2 h in broilers treated with L-NAME. 3. In Experiment 2, 180 Arbor Acres broilers were evenly divided into three groups: a control group (group C), and two groups exposed to low environmental temperatures and fed a 3, 3, 5-triiodothyronine (T3) supplemented diet alone (group A) or also including 100 ppm L-NAME (group B). 4. The PHS morbidity of group A was higher than for group C but lower than for group B. Plasma endothelin-1 was higher in broilers in groups A and B than in group C. Plasma NO was not significantly lower in broilers of group B when compared with those in group A. 5. The right/total ventricular weight ratio (RV/TV) and mean PAP were higher in groups A and B than in group C, and the RV/TV ratio increased one week earlier in group B than in group A. 6. These results suggest that L-NAME increases broiler PAP by inhibiting the endogenous synthesis of NO, leading to pulmonary hypertension, right ventricular hypertrophy and the increased morbidity of PHS in broilers.     

Supplementary dietary nitric oxide donor (sodium nitroprusside) or inhibitor (NG-nitro-l-arginine methyl ester) depressed growth performance and ovarian primordial and primary follicles in Japanese quail (Coturnix coturnix japonica) in a dose-dependent manner

1. The aim of the study was to determine the effects of dietary supplementation with sodium nitroprusside (SNP), a nitric oxide (NO) exogenous donor, and N^G-nitro-l-arginine methyl ester (l-NAME), a NO inhibitor, on growth performance, some biochemical parameters and ovarian primordial and primary follicles of quail.

2. A total of 480 Japanese quail (Coturnix coturnix japonica), one-day-old, including both males and females, were randomly allocated into one control group and 4 treatment groups each consisting of 96 birds. The control group was fed on the basal diet, whereas the experimental groups were fed on the basal diet supplemented with 50 mg SNP/kg, 200 mg SNP/kg, 50 mg l-NAME/kg or 200 mg l-NAME/kg.

3. In the group receiving 200 mg SNP/kg, BW was lower on d 28 and d 42 compared to the control group and body weight gain (BWG) was lower between weeks 2 and 4 compared to the control group. In the same group, BWG and feed consumption were lower compared with the control group. In the group receiving 200 mg l-NAME/kg, BW on d 42 and BWG were lower, whereas feed consumption and FCR was higher than in the control group.

4. In the groups supplemented with SNP at 50 and 200 mg/kg, serum total protein and albumin were higher than the control group; however, serum lipid profile, and liver and kidney enzymes were not affected by supplementation with SNP or l-NAME.

5. The numbers of ovarian primordial and primary follicles were greater in the group fed on the diet supplemented with 200 mg SNP/kg compared with the control group. Supplementation at 200 mg l-NAME/kg diet reduced the number of primary follicles compared to the controls, whereas the diameter of primordial and primary follicles increased.

6. In conclusion, supplementation with SNP and l-NAME depressed quail growth. Furthermore, the increase in NO following dietary supplementation with the NO-donor SNP delayed the growth process from primordial to primary and primary to secondary follicle transition in quail.

     

In ovo treatment with an aromatase inhibitor masculinizes postnatal hormone levels,abdominal fat pad content,and GH pulsatility in broiler chickens

Vorozole, a selective aromatase inhibitor, was administered in ovo to test the specific embryonic role of estrogen in conferring the sex distinction in GH release and body phenotype in broilers. On Day 6 of incubation, eggs were injected with saline or with different concentrations of vorozole. Postnatal blood samples were analyzed for T₃, T₄, GH, estradiol (E₂), and testosterone (T). At the age of 4 wk, control and vorozole-treated birds were cannulated, and serial blood samples were withdrawn every 10 min for 5 hr, wherein GH pulsatility characteristics were determined using deconvolution analysis. The proportional abdominal fat pad weight was reduced significantly in the treated groups, especially in female birds. The vorozole treatment increased plasma T₃, E₂, T, and GH concentrations, and decreased T₄. The frequency of the GH pulses was lower and the interval between the bursts (min) was higher in the vorozole-treated group, as were the mass secreted per burst (ng/ml), the amplitude (ng/ml/min) and the production rate (ng/ml/5 hr). In conclusion, early in ovo treatment with a potent aromatase inhibitor is able to increase the mean serum T₃ and GH concentration and masculinize the GH pulse pattern, resulting in an economically favorable decrease in abdominal fat pad content in male and female broilers at slaughter age.     

Endometrial nitric oxide synthase activity in mares susceptible or resistant to persistent breeding‐induced endometritis and the effect of a specific iNOS inhibitor in vitro

Emerging research suggests that the nitric oxide system may play a role in persistent breeding‐induced endometritis (PBIE) in the mare. Differences in uterine nitric oxide (NO) levels between mares susceptible or resistant to PBIE and a dose‐dependent inhibitory effect of NO on uterine contractility have been demonstrated. The objectives of this study were to investigate the difference in total nitric oxide synthase (NOS) activity of the endometrium between susceptible and resistant mares and the effect of a specific inducible nitric oxide synthase (iNOS) inhibitor on the endometrial NOS activity in vitro. Six susceptible and six resistant mares were selected based on preset criteria and the results of an intrauterine challenge with killed spermatozoa during oestrus. Endometrial biopsy samples were collected 24 hr post‐challenge and cultured at 37°C for 24 hr in L‐arginine supplemented minimum essential medium with or without a specific iNOS inhibitor (1,400 W dihydrochloride, 1 mM). The medium and the cultured endometrial tissue were collected after 24 hr of culture and assayed for NO and total protein, respectively. Total NO content of the medium, normalized to endometrial tissue wet weight or total protein, was used as a measure of endometrial NOS activity. Non‐parametric tests were applied for statistical analysis. Susceptible mares had significantly greater endometrial NOS activity than resistant mares. The iNOS inhibitor treatment significantly reduced NOS activity in endometrial samples derived from susceptible and resistant mares. These findings provide a basis for in vivo testing of specific iNOS inhibitors as preventative or therapeutic options for PBIE in mares.     

Effect of 7-nitroindazole, a selective neuronal nitric oxide synthase inhibitor, on parvalbumin immunoreactivity after cerebral ischaemia in the hippocampus of the Mongolian gerbil

Previous studies have demonstrated that a loss of parvalbumin-immunoreactive (PV-ir) neurones is observed in the hippocampus after transient cerebral ischaemia. However, whether the loss of parvalbumin (PV) immunoreactivity is related to the over-production of nitric oxide (NO) during cerebral ischaemia has not been evaluated. This study was designed to test the effect of 7-nitroindazole pre-treatment (7-NI, 50 mg/kg), a selective neuronal NO synthase inhibitor, on PV immunoreactivity and its cellular activity following forebrain ischaemia. PV-ir neurones in the hippocampus of the control group were widely distributed in the pyramidal cell layer and stratum oriens of CA1 and CA3, and the granular cell layer of dentate gyrus. 7-NI pre-treatment completely suppressed the reduction of PV immunoreactivity in CA1 that was observed in the ischaemia-induced group. Subsequently, 7-NI pre-treatment also protected against the structural loss of microtubule-associated protein 2 (MAP2) immunoreactivity in CA1 after ischaemic insult. In addition, the Fos-defined neuronal activity of PV-ir neurones was slightly increased by the 7-NI pre-treatment 3 h after ischaemia. Based on these data, we conclude that the neuronal toxicity of NO may be involved in the loss of PV-ir neurones after cerebral ischaemia.     

Ileal starch digestibility in growing broiler chickens fed on a wheat-based diet is improved by mash feeding, dilution with cellulose or whole wheat inclusion.

1. An experiment was conducted to study causes of low ileal starch digestibilities when broiler chickens were fed on wheat-based diets. Each of 5 cold-pelleted diets containing 771 g/kg DM wheat and one cold-pelleted diet containing 694 g/kg DM wheat were fed to 24 male broiler chickens in 8 cages from 10 to 21 d of age. 2. Feed intake and weight gain were significantly reduced when the wheat diet was crushed and fed in a mash form. 3. Ileal starch digestibility increased significantly from 0.79 to 0.95, 0.93 and 0.91, respectively, when the diet was crushed and fed in a mash form, was diluted with cellulose prior to pelleting, or when parts of the wheat were fed as whole grains. Correspondingly, random variation between individual birds was reduced. 4. These results indicate that an overload of wheat starch in the digestive tract may be the cause of poor digestibility for some broilers in a flock. Grinding of the wheat may also influence starch digestibility.     

Pharmacokinetic properties of toceranib phosphate (Palladia™, SU11654), a novel tyrosine kinase inhibitor,in laboratory dogs and dogs with mast cell tumors

Yancey, M. F., Merritt, D. A., Lesman, S. P., Boucher, J. F., Michels, G. M. Pharmacokinetic properties of toceranib phosphate (Palladia?, SU11654), a novel tyrosine kinase inhibitor, in laboratory dogs and dogs with mast cell tumors. J. vet. Pharmacol. Therap. 33 , 162–171. Toceranib phosphate (Palladia?, SU11654), an oral tyrosine‐kinase inhibitor, is under investigation for the treatment of mast cell tumors in dogs. The pharmacokinetics of toceranib phosphate has been characterized in dogs. Means of the following pharmacokinetic parameters were estimated following a 1.0 mg/kg i.v. dose to laboratory beagles: plasma clearance of 1.45 L/kg/h, volume of distribution of 29.7 L/kg, and terminal half‐life of 17.7 h. Following single oral doses of 3.25 mg/kg administered to laboratory beagles, mean C_max estimates ranged from 68.6 ng/mL to 112 ng/mL with t_max ranging from 5.3 h and 9.3 h postdose. Terminal half‐life was estimated at 31 h. Oral bioavailability was 76.9%. There were no statistically significant (P > 0.05) differences with any pharmacokinetic parameter due to fed/fasted state or with time during 13 weeks of every‐other‐day dosing at 3.25 mg/kg. Toceranib concentrations were proportional with dose over the range of 2.0 to 6.0 mg/kg. The pharmacokinetics of toceranib in client‐owned dogs of a variety of pure and mixed breeds with mast cell tumors was similar to that in healthy laboratory dogs. In summary, toceranib phosphate exhibited moderate clearance, a high volume of distribution, and a moderate elimination half‐life. After a single oral dose at 3.25 mg/kg, the concentration vs. time curve showed broad, sustained exposure with measurable concentrations for more than 48 h. These pharmacokinetic parameters support every‐other‐day administration of toceranib phosphate at an initial dose of 3.25 mg/kg for the treatment of mast cell tumors in dogs.