Fabio Ferrante

Long term effects of bosentan treatment in adult patients with pulmonary arterial hypertension related to congenital heart disease (Eisenmenger physiology) : safety, tolerability, clinical, and haemodynamic effect

Background: Oral bosentan is an established treatment for pulmonary arterial hypertension (PAH). Objective: To evaluate safety, tolerability, and clinical and haemodynamic effects of bosentan in patients with PAH related to congenital heart disease (CHD). Patients: 22 patients with CHD related PAH (8 men, 14 women, mean (SD) age 38 (10) years) were treated with oral bosentan (62.5 mg×2/day for the first 4 weeks and then 125 mg×2/day). Main outcome measures: Clinical status, liver enzymes, World Health Organisation (WHO) functional class, resting oxygen saturations and 6-min walk test (6MWT) were assessed at baseline and at 1, 3, 6, and 12 months. Haemodynamic evaluation with cardiac catheterisation was performed at baseline and at 12 month follow-up. Results: 12 patients had ventricular septal defect, 5 atrioventricular canal, 4 single ventricle, and 1 atrial septal defect. All patients tolerated bosentan well. No major side effects were seen. After a year of treatment, an improvement was seen in WHO functional class (2.5 (0.7) v 3.1 (0.7); p<0.05), oxygen saturation at rest (87 (6%) v 81 (9); p<0.001), heart rate at rest (81 (10) v 87 (14) bpm; p<0.05), distance travelled in the 6MWT (394 (73) v 320 (108) m; p<0.001), oxygen saturation at the end of the 6MWT (71 (14) v 63 (17%); p<0.05), Borg index (5.3 (1.8) v 6.5 (1.3); p<0.001), pulmonary vascular resistances index (14 (9) v 22 (12) WU m2; p<0.001), systemic vascular resistances index (23 (11) v 27 (10) WU.m2; p<0.01), pulmonary vascular resistances index/systemic vascular resistances index (0.6 (0.5) v 0.9 (0.6); p<0.05); pulmonary (4.0 (1.3) v 2.8 (0.9) l/min/m2; p<0.001) and systemic cardiac output (4.2 (1.4) v 3.4 (1.1) l/min/m2; p<0.05). Conclusions: Bosentan was safe and well tolerated in adults with CHD related PAH during 12 months of treatment. Clinical status, exercise tolerance, and pulmonary haemodynamics improved considerably.

Age-dependent nerve cell loss in the brain of Sprague-Dawley rats: effect of long term acetyl-L-carnitine treatment.

The age dependent loss of nerve cells was investigated in 22 brain areas from young (3 month), adult (13 month) and old (25 month) Sprague-Dawley rats. As in previous studies, we observed an age-related neuronal loss primarily in the archicortex and in the hippocampus and in other subcortical structures (amigdaloid nucleus, pontine nuclei, cerebellar cortex). In sensory areas of cerebral cortex the neuronal loss was less marked. The effect of a 6 month treatment with acetyl-L-carnitine (ALCAR) on the number of nerve cells in the same brain areas was also investigated. ALCAR treatment began when the rats were aged 16 months. ALCAR treatment was able to counteract the age-dependent decrease in nerve cell number primarily in the temporal and occipital cortical areas, in the archicortex and hippocampus. The above findings suggest that long term ALCAR treatment may be effective in slowing down the age-related nerve cell loss in some rat brain areas.

Acetylcholinesterase-containing nerve fibers in the dura mater of guinea pig, mouse, and rat

The cholinergic innervation of the dura mater in the guinea pig, mouse and rat has been investigated. The dura mater is provided with a cholinergic innervation. After short incubation times (4–6 hours) we found acetylcholinesterase (AChE)-containing nerve fibers in close relationship with the main meningeal blood vessels. After longer incubation times, AChE-containing nerve fibers appear also in the meningeal tissue proper. Chemical sympathectomy, performed with the neurotoxin 6-hydroxydopamine, does not seem to interfere with the pattern of the dural cholinergic innervation. The findings are discussed.

Cholinergic nerves in the human liver

SummaryThe cholinergic innervation of the human liver was studied. Slices (150–200μm thick) of human liver and of the greater hepatic blood vessels (hepatic artery and vein, portal vein) were incubated in a solution of 6-hydroxydopamine (6-HDA) in order to obtain a selective degeneration of adrenergic nerves. Controls were prepared from samples incubated with buffer alone. The slices were cut on a cryostat into 15–20μm thick sections and processed for the histochemical detection of cholinesterases.Cholinergic nerve fibres innervate the extra hepatic and the intrahepatic branches of the hepatic artery, the portal vein as well as the hepatic vein. Fewer cholinergic fibres innervate the hepatocytes and the hepatic sinusoids. The 6-HDA treatment does not seem to alter the pattern of the cholinergic innervation of the liver. The findings indicate the presence of a cholinergic parasympathetic innervation in the human liver.

Prognostic factors in severe pulmonary hypertension patients who need parenteral prostanoid therapy: The impact of late referral

BACKGROUND Oral drugs have made the treatment of pulmonary hypertension (PH) feasible in non-expert centers, which could delay patient access to prostanoid therapy. METHODS Fifty-seven consecutive patients with precapillary PH received a prostanoid in our center. Data at prostanoid initiation included modality of center referral, medical history, New York Heart Association [NYHA] class, exercise capacity, echocardiographic parameters, and hemodynamics. RESULTS Overall survival at 1, 2, and 3 years was 85%, 69%, 55%, respectively. Non-survivors had worse NYHA class III/IV (17/12) than survivors (27/1; p < 0.01) and exercise capacity on 6-minute-walk distance (254 ± 114 vs 354 ± 91 meters; p < 0.01). Non-survivors were more frequently referred on oral therapy (83% vs 36%; p < 0.01) and had a higher rate of urgent prostanoid treatment (69% vs 17%; p < 0.0001). Multivariate analysis (hazard ratio [95% confidence interval]) found the independent prognostic factors were urgent prostanoid therapy (2.0 [1.1-3.9]) and NYHA class (3.5 [1.5-8.2]). Survivors had a significant response to prostanoid, improving NYHA class from 2.8 ± 0.4 to 2.3 ± 0.5 (p = 0.002), 6-minute walk distance from 354 ± 91 to 426 ± 82 meters (p = 0.0001), and pulmonary hemodynamics (pulmonary artery pressure from 56 ± 13 to 44 ± 18 mm Hg [p < 0.05]; cardiac index from 2.0 ± 1.2 to 3.1 ± 1.2 liters/min/m(2) [p = 0.002], and pulmonary vascular resistance from 17 ± 10 to 8 ± 6 WU [p = 0.001]). CONCLUSIONS Referral of patients on oral treatment to a tertiary PH center is delayed and significantly affects prognosis.

Enzyme histochemistry of the choroid plexus in old rats

The influence of ageing on the metabolic profile of lateral ventricle choroid plexus epithelial cells from young (3-month-old) and aged (26-month-old) male Wistar rats was studied using enzyme histochemical techniques. The following enzymatic activities related to energy transduction were examined: lactate-(LDH) and succinate- (SDH) dehydrogenases; NADH2-tetrazolium reductase (NADHD) and alpha-glycerophosphate-dehydrogenase (GPDH). The intensity of enzymatic staining within single choroid plexus epithelial cells from young and old animals was assessed microphotometrically. In the choroid plexus epithelial cells of young rats NADHD was the enzymatic activity more heavily stained; cell levels of LHD and GPDH were approximately the same and SDH reactivity was less intense. In old age LDH was reduced by 9.3%, SDH was reduced by 26.1%, NADHD was reduced by 8.6% and GPDH was reduced by 3.6%. The possibility that impaired energy transduction mechanisms at the level of choroid plexus epithelium in old age may influence functional activity of the choroid plexus is discussed.

Age-dependent changes in the expression of dopamine receptor subtypes in human peripheral blood lymphocytes

The pharmacological profile and the density of dopamine D3 and D5 receptor subtypes expressed by human peripheral blood lymphocytes of subjects of different ages (ranging from 20 to 75 years) were assessed using radioligand binding techniques. Dopamine D3 receptor was assayed with [3H]7-hydroxy-N,N-di-n-propyl-2-aminotetraline ([3H]7-OH-DPAT) as a ligand. Dopamine D5 receptor was assayed using [3HIR]-(+)-(-chloro-2,3,4,5, tetrahydro-5-phenyl-1H-3-benzazepin-al-hemimaleate) ([3H]SCH 23390) as a ligand. The affinity and the pharmacological profile of [3H]7-OH-DPAT and [3H]SCH 23390 at dopamine D3 and D5 receptor, respectively, were similar in subjects of different ages. The density of dopamine D3 receptor binding sites was slightly decreased in subjects of 30-39 years in comparison with younger individuals. A remarkable loss of dopamine D3 receptor was then found between 40 and 49 years of age in comparison with younger subjects. A further slight decrease was noticeable between 50 and 59 years of age. The number of [3H]7-OH-DPAT binding sites was then stabilized after 60 years of age. The density of dopamine D5 receptor binding sites did not show age-dependent changes. The above findings indicate the occurrence of a decline in the density of lymphocyte dopamine D3 but not D5 receptor between adult and mature subjects. The possibility that dopamine D3 receptor assay in peripheral blood lymphocytes may represent a tool for investigating dopamine receptor function in aging and age-related neurological disorders is discussed.

Microanatomical changes in the frontal cortex of aged rats: effect of L-deprenyl treatment.

The present study was designed to assess whether treatment with L-deprenyl has any effect on the age-related microanatomical changes in the rat frontal cortex. Male Sprague-Dawley rats of 19 months of age were treated until the 24th month with an oral daily dose of 1.25 mg/kg or of 5 mg/kg of L-deprenyl. Eleven-month-old untreated rats were used as an adult reference group. The density of nerve cell profiles and of glial fibrillary acidic protein-(GFAP) immunoreactive astroglial profiles, lipofuscin accumulation within the cytoplasm of pyramidal neurons, and MAO-B reactivity were assessed. A decreased density of nerve cell profiles and an increased density of astroglial profiles as well as augmented lipofuscin deposition and MAO-B reactivity were observed in the frontal cortex of rats of 24 months in comparison with 12-month-old animals. In the frontal cortex of rats treated with 5 mg/kg/day L-deprenyl, which is a dose inhibiting MAO-B activity, the density of nerve cell and GFAP-immunoreactive astrocyte profiles is increased and decreased respectively in comparison with age-matched untreated subjects. Lipofuscin deposition is reduced. The lower dose of L-deprenyl (1.25 mg/kg/day) which did not affect MAO-B activity, decreased lipofuscin deposition but was without effect on the density of nerve cell or GFAP-immunoreactive astrocyte profiles. The above findings suggest that treatment with L-deprenyl is able to counter some microanatomical changes occurring in the frontal cortex of aged rats. Some of these effects are probably not related to the inhibitory MAO-B activity of the compound.

Histochemical studies on the autonomic innervation of the femoral artery and vein

SummaryThe innervation of the femoral artery and vein in the dog was studied using: a) glyoxylic acid fluorescence (for the histochemical localization of adrenergic nerve fibers); b) acetylcholinesterase (for the histochemical localization of cholinergic nerve fibers).—Andrenergic nerve fibers and related terminals in both vessels are confined to the adventitial-medial transitional zone and the outer layers of the media. Acetylcholinesterase containing nerve fibers are localized in the adventitial-medial transitional zone of the femoral artery. These findings may indicate the presence of a cholinergic vosodilatator system in the control of the circulation in the femoral artery.

Effects of Long-Term Hydergine® Administration on Lipofuscin Accumulation in Senescent Rat Brain

The effects of ageing and of 6 months of Hydergine treatment on lipofuscin deposition within the cytoplasma of pyramidal neurons of rat prefrontal cortex, hippocampus (fields CA1 and CA3) and of Purkinje neurons were assessed microfluorimetrically. No lipofuscin autofluorescence was detected in the nerve cell populations of 3-month-old rats, but lipopigment had accumulated in nerve cell bodies of 16-month-old animals and increased significantly thereafter in rats of 22 months of age. In 22-month-old rats, Hydergine administration (0.6 and 1 mg/kg p.o.) started at 16 months caused a significant dose-related decrease in lipofuscin accumulation within the cytoplasm of the various kinds of nerve cells examined.