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D Curti

Publications and source records attributed to D Curti.

At least 37 records · Page 2Linked to original sources

Age-related modification of enzyme activities in synaptosomes isolated from rat cerebral cortex.

In synaptosomes from rat cerebral cortex, the potential catalytic activity of some enzymes related to energy metabolism--namely, phosphofructokinase and citrate synthase--is not affected by aging. In contrast, the maximum velocity (Vmax) of cytochrome oxidase and of pyruvate dehydrogenase decreases in aged rats. A marked increase is found in the Vmax of glucose 6-phosphate dehydrogenase in aged rats and could be related to the availability of NADPH for antiperoxidative processes. Pretreatments of experimental animals with certain drugs were done to investigate the plasticity of enzyme proteins during aging. Papaverine, which acts on macrocirculation, is ineffective, but delta-yohimbine acting on microcirculation and metabolism and almitrine acting on oxygen availability both could interfere with the potential activity of some enzymes. However, their influence differs with the age of the rats.

Aging↗

Effect of Ca2+-homopantothenate and mild hypoxia on some enzyme activities evaluated in subcellular fractions from different rat brain regions.

The effect of Ca2+-homopantothenate (HOPA) treatment (250 mg/kg for 5 d) has been studied by evaluating the specific activity of enzymes related to: glycolytic pathway (hexokinase, phosphofructokinase, pyruvate kinase, lactate dehydrogenase), tricarboxylic acid cycle (citrate synthase, malate dehydrogenase), mitochondrial electron transfer chain (succinate dehydrogenase, cytochrome oxidase), NADH redox state (NADH cytochrome c reductase), acetylcholine metabolism (acetylcholinesterase), and glutamate metabolism (glutamate dehydrogenase). The enzymatic activity assays were performed on homogenate in toto, nonsynaptic mitochondria and synaptosomes isolated from: cerebral cortex, hippocampus, striatum, hypothalamus, medulla oblongata, and cerebellum of normoxic rats and rats submitted to intermittent normobaric hypoxia (90:10, N2:O2). In normoxic rats, HOPA was unable to induce any modification. Hypoxia per se induced a decrease in the activity of synaptosomal cytochrome oxidase in cerebral cortex, hippocampus, and cerebellum.

Acetylcholinesterase↗

Effect of aging and acetyl-L-carnitine on energetic and cholinergic metabolism in rat brain regions.

The effect of aging and subchronic treatment with acetyl-L-carnitine (50 mg/kg per day) was studied on mitochondrial bioenergetics and cholinergic metabolism in non-synaptic mitochondria and synaptosomes isolated from cerebral cortex, hippocampus and striatum of rats aged 4, 11 and 18 months. Respiratory activity and cytochrome oxidase specific activity were unaffected by aging in non-synaptic mitochondria. In synaptosomes, pyruvate dehydrogenase, choline acetyltransferase and acetylcholinesterase specific activity remained unchanged, but the high-affinity choline uptake decreased in cerebral cortex and striatum of 18-month-old rats. Acetyl-L-carnitine treatment increased the high-affinity choline uptake in cerebral cortex of 18-month-old rats. The treatment caused also an increase in cytochrome oxidase activity in all the three cerebral regions and in choline uptake in the hippocampus, parameters that were not directly affected by aging processes.

Acetylcarnitine↗

Oxidative metabolism of nonsynaptic mitochondria isolated from rat brain hippocampus: a comparative regional study.

Nonsynaptic mitochondria isolated from rat brain hippocampus were compared with those obtained by means of the same preparative procedure from cerebral cortex and striatum. Protein recovery, marker enzyme activities (lactate dehydrogenase, citrate synthase, and acid phosphatase), state 4 respiration, and response to hypoosmotic shock showed no difference among the three cerebral regions, suggesting homogeneous behavior during the subfractionation procedure. Cholinergic markers--choline acetyltransferase, acetylcholinesterase activities, and high-affinity choline uptake--evaluated on synaptosomes showed the classic regional pattern with an enrichment in the striatum (striatum much greater than hippocampus). The coupling state of the mitochondrial fractions was maintained (respiratory control ratios ranging from 3.62 to 5.08 with glutamate + malate as oxidizable substrates), showing a metabolic competence sufficient to perform metabolic studies. Regional differences were found in state 3, uncoupled state of respiration, and cytochrome oxidase activity. Hippocampus showed the lower values (hippocampus less than striatum less than cortex). A possible role of this lower capacity of mitochondrial energy metabolism in determining the sensitivity of hippocampal neurons to ischemia or epileptic seizures is suggested.

Acetylcholinesterase↗

Phenobarbital and 6-aminonicotinamide effect on cerebral enzymatic activities related to energy metabolism in different rat brain areas.

The effect of phenobarbital (100 mg/kg i.p.) and 6-aminonicotinamide (6AN) (35 mg/kg i.p.) on enzyme activities related to energy transduction was investigated on the homogenate "in toto", non-synaptic mitochondrial fraction and synaptosomal fraction isolated from different rat brain areas (cerebral cortex, hippocampus, hypothalamus, striatum, and medulla oblongata). 6AN treatment decreased: phosphofructokinase in all the areas tested; lactate dehydrogenase on the homogenate "in toto" in striatum and hypothalamus, and on the synaptosomal fraction in cerebral cortex and corpus striatum; succinate dehydrogenase on non-synaptic mitochondrial fraction in hippocampus and striatum. Finally, aspartate aminotransferase was increased on non-synaptic mitochondrial fraction in striatum and medulla oblongata. Phenobarbital treatment induced an increase of total NADH cytochrome c reductase on mitochondrial fraction in hippocampus and hypothalamus, and a decrease of cytochrome oxidase activity on non-synaptic mitochondrial fraction in hypothalamus and medulla oblongata.

6-Aminonicotinamide↗

Brain enzyme adaptation to mild normobaric intermittent hypoxia.

The adaptation to repeated periods of intermittent normobaric hypoxia (oxygen:nitrogen = 10:90, 12 hr daily for 5 days) of some specific enzymatic activities related to energy metabolism has been observed in different rat brain areas (cerebral cortex, hippocampus, corpus striatum, hypothalamus, cerebellum, and medulla oblongata). The evaluation of the maximum rate (Vmax) of the enzymes was carried out on: the homogenate "in toto," the nonsynaptic mitochondrial fraction, and the crude synaptosomal fraction. The adaptation to intermittent normobaric hypoxic exposure was characterized by significant modifications of some enzyme activities in the homogenate "in toto" (decrease of hexokinase activity in cerebellum), in the nonsynaptic mitochondrial fraction (increase of succinate dehydrogenase activity in corpus striatum and decrease of cytochrome oxidase activity in cerebral cortex), and, particularly, in the synaptosomal fraction (decrease of cytochrome oxidase activity in cerebral cortex, hippocampus, corpus striatum, and cerebellum, and decrease of malate dehydrogenase and lactate dehydrogenase activity in cerebellum). The adaptation to normobaric intermittent hypoxia differs according to the brain area, subcellular fraction, and enzyme activity tested.

Adaptation, Physiological↗

Nuclear and mitochondrial DNA synthesis and energy metabolism in primary rat glial cell cultures.

DNA synthesis in nuclei and mitochondria purified from serum-supplemented rat glial cell cultures at different days after plating was studied. Furthermore in mitochondria, some enzymatic activities related to energy transduction (citrate synthase, malate dehydrogenase, total NADH-cytochrome c reductase, cytochrome oxidase and glutamate dehydrogenase) were measured. For DNA labeling [methyl-3H]thymidine was added to the culture medium at different days after plating. During the culture times studied the specific activity of total, nuclear, and mitochondrial DNA decreased from 8 days in vitro (DIV) to 21 DIV and increased at 30 DIV. The specific activity of nuclear DNA was always higher than that of mitochondrial DNA. The specific activity of the above mentioned mitochondrial enzymes increased from 8 DIV up to 21 DIV and decreased at 30 DIV, suggesting a relationship between the energy metabolism and the differentiation of glial cells in culture.

Animals↗

Influence of intermittent hypoxia and pyrimidinic nucleosides on cerebral enzymatic activities related to energy transduction.

The effect of intermittent normobaric hypoxia and of biological pyrimidines (uridine and cytidine) on the specific activities of some enzymes related to cerebral energy metabolism were studied. Measurement were carried out on the following: homogenate in toto; purified mitochondrial fraction; crude synaptosomal fraction, in different areas of rat brain: cerebral cortex, hippocampus, corpus striatum, hypothalamus, cerebellum, and medulla oblongata. Intermittent normobaric hypoxia (12 hours daily for 5 days) caused modifications of the enzyme activities in the homogenate in toto (decrease of hexokinase in cerebellum; increase of pyruvate kinase in medulla oblongata), in the purified mitochondrial fraction (increase of succinate dehydrogenase in the corpus striatum) and in the crude synaptosomal fraction (decrease of cytochrome oxidase activity in cerebral cortex, hippocampus, and cerebellum; decrease of malate dehydrogenase in hippocampus and cerebellum; decrease of lactate dehydrogenase in cerebellum). Daily treatment with cytidine or uridine altered some enzyme activities either affected or unaffected by intermittent hypoxia.

Animals↗

Kinetics of irreversible inhibition of choline transport in synaptosomes by ethylcholine mustard aziridinium.

Ethylcholine mustard aziridinium (ECMA) inhibits choline transport in synaptosomes at a half-maximal concentration of about 20 microM. The rate of inhibition falls off rapidly after 10 min and the concentration dependency reaches a plateau at about 100 microM. The inhibition is not removed by washing the synaptosomes, and choline and hemicholinium-3 protect the carrier against attack by the mustard. Choline efflux, particularly that stimulated by choline in the medium (transactivation) is also inhibited by the aziridinium compound. Similarly choline influx activated by preloaded internal choline is inhibited by ECMA. The mustard can enter the synaptosomes in an active form but most of the carrier is alkylated when facing the outside. Prior depolarization of the synaptosomes causes an increase in the rate of inhibition by ECMA which is proportionally about the same as the increase in choline influx also caused by depolarization. At low ECMA concentrations the rate of inhibition is that of a first-order reaction with the carrier but at high ECMA concentrations the translocation of the carrier to the outward-facing conformation controls the rate of inhibition. Using a model of choline transport with some simplifying assumptions it is possible to estimate the amount of carrier; cholinergic synaptosomes carry about six times the concentration of carrier found in noncholinergic ones. In noncholinergic synaptosomes the carrier faces predominately out, the reverse in cholinergic ones. The rate constant of carrier translocation is increased by combination with choline some six- to sevenfold to about 3.5 min-1. The rate constant of ECMA attack on the carrier is about 440 M-1 sec-1.

Animals↗

Effect of prolonged and intermittent hypoxia on some cerebral enzymatic activities related to energy transduction.

The adaptation to repeated, alternate normobaric hypoxic and normoxic exposures (12 h/day, for 5 days) and to pharmacological treatment was evaluated by studying the specific activities of some enzymes related to cerebral energy metabolism. Measurements were carried out on (a) the homogenate in toto, (b) the purified mitochondrial fraction, and (c) the crude synaptosomal fraction in different areas of rat brain--cerebral cortex, hippocampus, corpus striatum, hypothalamus, cerebellum, and medulla oblongata. The adaptation to intermittent normobaric hypoxic-normoxic exposures was characterized by significant modifications of some enzyme activities in synaptosomes (decrease of cytochrome oxidase activity in the hippocampus, corpus striatum, and cerebellum; decrease of malate dehydrogenase activity in the cerebellum) and in the purified mitochondrial fraction (increase of succinate dehydrogenase activity in the corpus striatum). Daily treatment with three doses of naftidrofuryl (10, 15, and 22.5 mg/kg i.m.) modified some enzyme activities affected or unaffected by intermittent hypoxia and, particularly, decreased acetylcholinesterase activity.

Animals↗

Effect of hypoxia and pharmacological treatment on some enzyme activities in dog brain areas.

The effects of nicergoline on changes in enzymatic activities induced by hypoxia and post-hypoxic recovery were studied in various brain areas of young-adult and mature Beagle dogs. In different fractions (homogenate in toto, purified mitochondria, crude synaptosomes, SM1 and SM2 synaptic mitochondria) the maximal rate (Vmax) was investigated of the more representative enzymatic activities of: a) glycolysis, b) Krebs' cycle, c) electron transfer chain, d) amino acid and acetylcholine metabolism, e) lysosomal function. The physiopathological conditions caused alterations in different enzymatic activities depending on the area and subfraction investigated. Nicergoline tended to antagonize some of these alterations. Its action was mainly on non-synaptic mitochondria by a "braking" effect on some key enzyme activities of mitochondrial metabolism (i.e. citrate synthase, cytochrome oxidase and glutamate dehydrogenase) which suggests a sparing action in the brain.

Aging↗

[Variability of the radiologic characteristics of osteoid osteoma].

The authors have studied the anatomical-radiographic correlations in 55 patients with osteoid osteoma. On the basis of the results gathered we feel able to state that the variability of radiologic features of osteoid osteoma (nidus surrounded by a zone of sclerosis) depends on the site of the lesion and on the advanced stage of the neoplastic tissue.

Adolescent↗

Influence of age upon the cerebral metabolic changes induced by acute hypoxia on the synaptosomes from dog brain.

The synaptosomal fraction obtained from the motor area of the cerebral cortex of normocapnic, normoxic or hypoxic "young adult," "mature" and "senescent" beagle dogs is incubated and analyzed for : ATP, ADP, AMP, creatine phosphate, pyruvate and lactate. The data are compared with those obtained from the whole controlateral cortical motor area, by the surface freezing technique. After hypoxic hypoxia /15 min; PaO2 = 17-19 mm Hg), the metabolite contents and ratios are differently affected by ageing when the evaluations are performed in the incubated synaptosomal preparation or in the controlateral whole cerebral tissue. In fact, ageing does not affect so much the cerebral changes that occur in the overall energetic state during the hypoxic assault in vivo, but rather those that the synaptosomes remember the tend to reverse during the subsequent incubation in vitro. The protective action of several drugs on the synaptosomal phosphorylation state is tested. Phenobarbital shows a quite broad, age-independent spectrum of action. (-)Eburnamonine and dihydroergocristine exhibits a more limited, age-dependent effectiveness, but are devoid of anesthetic action. Papaverine proves unable to affect the tested biochemical parameters.

Adenine Nucleotides↗

Effects of ischemia and pharmacological treatment on subcellular fractions from neonatal rat brain.

The effects of complete ischemia and of in vivo pharmacological treatment with trimetazidine were studied on some enzymatic activities related to energy transduction: lactate dehydrogenase for anaerobic glycolysis; citrate synthase and malate dehydrogenase for the Krebs' cycle; total NADH-cytochrome c reductase and cytochrome oxidase for the electron transport chain; glutamate dehydrogenase for amino acid metabolism and acetylcholine esterase for acetylcholine metabolism. These enzymatic activities were evaluated in brains of 10-day-old rats, at three different subcellular levels: homogenate in toto, purified mitochondrial fraction, crude, synaptosomal fraction. Complete normothermic post-decapitative ischemia of 30 min duration increased the activity of cytochrome oxidase in the homogenate in toto and increased the activities of citrate synthase and malate dehydrogenase in the purified mitochondrial fraction, the activities of the enzymes evaluated in the crude synaptosomal fraction being unaffected. The i.p. treatment with trimetazidine (at the dose level of 50 mg . kg-1) was without any significant effect on the tested enzymatic activities.

Animals↗

Metabolic changes induced by acute hypoxia on the synaptosomes from dog brain.

Synaptosomal preparations from the motor area of the cerebral cortex of normocapnic, normoxic or hypoxic untreated beagle dogs and phenobarbital-, papaverine-, and (-) eburnamonine-treated dogs were incubated for 10 min at 24 degrees C and analyzed for ATP, ADP, AMP creatine phosphate, pyruvate, and lactate. The data were compared with those obtained from the whole controlateral cortical motor area, by the surface-freezing technique. Both during normoxia and after hypoxic hypoxia (15 min, at PaO2 equal to 17-19 mm Hg) the metabolite contents and ratios were very different in the incubated synaptosomal preparations and in the whole cerebral tissues. As concerns the drug treatment, papaverine was always inactive, while (1) eburnamonine increased the synaptosomal phosphorylation state in hypoxic dogs, being ineffective on the glycolytic metabolites evaluated. Phenobarbital increased the synaptosomal phosphorylation state both in normoxic and in hypoxic animals, and was effective also on the glycolytic metabolites studied.

Adenine Nucleotides↗

Cerebral enzymatic activities during chronic hyperammonemia and treatment with S-adenosyl-L-methionine, adenosine and methionine in the rat.

The effect of the chronic intramuscular administration of some agents related to the S-adenosyl-L-methionine system on the hyperammonemia syndrome was evaluated. This experimental syndrome was induced in the rat by intraperitoneal administration of high doses of ammonium acetate (33, 100 and 300 mg/kg/day, 6 days a week for 80 days) followed by the assay of the activities of some cerebral enzymes involved in energy transduction. The enzymatic activities studied in the homogenate and in the mitochondrial fractions of brain tissue were: lactate dehydrogenase, citrate synthase, malate dehydrogenase, total NADH-cytochrome c reductase and cytochrome oxidase. All three doses of ammonium acetate induced significant modifications in the cerebral enzymatic activities. These doses reduced the activity of the total NADH-cytochrome c reductase both in the homogenate and in the mitochondrial fraction. On the other hand the activity of malate dehydrogenase was reduced limited to the two lower doses in the homogenate only. The simultaneous daily treatment (i.m.) with equimolar doses of substances involved in the S-adenosyl-L-methionine system (adenosine, methionine and S-adenosyl-L-methionine) did not cause any significant modification of the cerebral enzymatic activities associated with the administration of ammonium acetate at the three dose levels, thus confirming our previous results.

Adenosine↗

Effect of vincamine on some enzymatic activities from various areas of the beagle dog cerebral cortex.

The effect of a chronic (3 months) treatment with vincamine on the enzymatic activities related to energy transduction was studied on several areas of the cerebral cortex of dog brain. About enzymatic activities of the four different cortical areas, in controls, no difference was observed between the enzymatic activities evaluated in the crude mitochondrial fraction, with regard to both the tricarboxylic acid cycle (citrate synthase, malate dehydrogenase) and the electron transport chain (total NADH-cytochrome c reductase, cytochrome oxidase). On the contrary, in the homogenate, lactate dehydrogenase, malate dehydrogenase and acetylcholine esterase showed different maximal activities. In the crude mitochondrial fraction the intravenous treatment with the three different doses of vincamine failed to cause any significant change as compared to controls. On the contrary, with regard to the enzymatic activities evaluated in the homogenate in toto, the analysis of variance revealed an effect on cytochrome oxidase at the dose of 3 mg/kg intravenously.

Animals↗