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Biomedical subjects

P Marini

Publications and source records attributed to P Marini.

At least 37 records · Page 2Linked to original sources

Expression of hexokinase mRNA in human hippocampus.

The expression of hexokinase messenger RNA was evaluated in human hippocampus using in situ hybridization technique. The message showed an uneven distribution with high levels present in the granular cell layer of the dentate gyrus and CA3 region. The detection of specific transcripts was also observed in the lateral geniculate nucleus, the dentate polymorphic cell layer and the parahippocampal gyrus. The data suggest that, in the hippocampus, the expression of hexokinase is higher in neurons than in glial cells and that the rate of glucose metabolism may display considerable variations in the different subregions of this area.

Adult↗

Radiation-induced apoptosis in human lymphocytes and lymphoma cells critically relies on the up-regulation of CD95/Fas/APO-1 ligand.

Ionizing radiation is known to induce death by apoptosis in malignant and nonmalignant human lymphocytes. The mechanism which initiates the induction of apoptosis has not been identified. Here we demonstrate that a radiation-induced up-regulation of CD95/Fas/APO-1 ligand (CD95-L) is involved in the induction of apoptosis in lymphocytes and lymphoma cells. Using antibodies against CD95-L, we detected a rapid and persistent up-regulation of CD95-L in Jurkat cells and blood lymphocytes after irradiation. Blocking of interactions between CD95/Fas/APO-1 and CD95-L with inhibitory antibodies reduces apoptosis after irradiation. The hypothesis that an interaction between CD95 and CD95-L is involved in radiation-induced apoptosis is supported by the finding that only peripheral blood lymphocytes that are stimulated with interleukin 2 for 5 days and thus are rendered sensitive to CD95-L show pronounced levels of apoptosis in response to low radiation doses, whereas resting peripheral blood lymphocytes show very low levels of apoptosis after treatment with CD95-L and radiation. The finding that the caspase FLICE is degraded proteolytically after irradiation and the observation of a radiation-induced CD95 receptor capping confirm the functional activation of the CD95 receptor.

Apoptosis↗

Emotional arousal and phobia in transient global amnesia.

OBJECTIVE: To evaluate the role of emotionally stressful or phobogenic events and phobic personality traits in transient global amnesia (TGA). DESIGN: Case-control study. SETTING: Tertiary care center. PATIENTS: Fifty-one case patients with TGA (mean +/- SD age, 62.7 +/- 6.7 years) compared with 51 control patients with transient ischemic attacks (mean +/- SD age, 63.8 +/- 6.7 years). MAIN OUTCOME MEASURES: Precipitant factors, life events, and phobic attitudes. RESULTS: Of the 25 TGA attacks that were triggered by a precipitant, 11 were possibly related to emotionally stressful or phobogenic situations. On a scale that measured phobic attitudes, the case patients with TGA scored significantly higher than the control patients with transient ischemic attacks (mean +/- SD total score, 15.21 +/- 11.0 vs 4.41 +/- 5.2; P < .001 by corrected analysis of variance for age, sex, and education). The amount of stressful live events in the year that preceded the attack did not differ between the case patients with TGA and the control patients with transient ischemic attacks. CONCLUSION: The results support the hypothesis that emotional arousal and phobia are involved in TGA.

Aged↗

Interaction of thiocolchicoside with [3H]strychnine binding sites in rat spinal cord and brainstem.

Radioreceptor binding assays and receptor autoradiography were used to investigate the activity of thiocolchicoside on strychnine-sensitive binding sites in rat brain and spinal cord using [3H]strychnine as a ligand. Thiocolchicoside displaced the binding of [3H]strychnine with an affinity similar to that of unlabeled glycine, and showed a Hill coefficient and proportionality parameter (P) less than unity. The activity of thiocolchicoside toward [3H]strychnine binding sites was confirmed in autoradiographic studies. The results suggest that thiocolchicoside behaves as an allosteric compound acting on the strychnine-sensitive glycine receptor in rat brainstem and spinal cord, and that this may provide a possible mechanism for the myorelaxant activity of this colchicoside derivative, the first clinically useful drug acting on this receptor.

Animals↗

Changes in protein kinase C and its presynaptic substrate B-50/GAP-43 after intrauterine exposure to methylazoxy-methanol, a treatment inducing cortical and hippocampal damage and cognitive deficit in rats.

The involvement of protein kinase C (PKC)-dependent processes in adaptive and plastic changes underlying neuronal plasticity was tested in an in vivo animal model characterized by targeted cellular ablation of cortical and hippocampal neurons, cognitive impairment and lack of induction of long-term potentiation. [3H]Phorbol ester binding performed on brain slices revealed a 67.4 and 35.0% increase in membrane-bound protein kinase C in the cortex and hippocampus respectively of rats treated with methylazoxy-methanol acetate compared with saline-treated control rats, and there was no modification in the expression of mRNAs of different protein kinase C isozymes. In situ phosphorylation experiments performed with 32Pi-labelled synaptosomes from the affected areas demonstrated that the phosphorylation of the nervous tissue-specific presynaptic membrane-associated protein kinase C substrate B-50/GAP-43 was increased by 51.4 and 44.8% in cortex and hippocampus respectively. Western blot analysis of protein kinase C in synaptosomal cytosol and membrane fractions prepared from cortex and hippocampus showed an increased proportion of protein kinase C in the membrane compartment in treated animals, but no change in the total synaptosomal protein kinase C activity. Our data are consistent with increased activity of presynaptic protein kinase C and predict a sustained increase in glutamate release in methylazoxy-methanol-treated rats.

Animals↗

Expression of neuronal acetylcholine nicotinic receptor alpha 4 and beta 2 subunits during postnatal development of the rat brain.

The expression of the alpha 4 and beta 2 subunits of neuronal nicotinic acetylcholine receptors (nAChRs) was studied in developing rat brain using in situ hybridization. The levels of both transcripts were already high at birth in cerebral cortex, medial habenula, CA1/CA3 regions of the hippocampus and several thalamic nuclei. In general, the beta 2 subunit showed a higher density of hybrids than the alpha 4. Beta 2 expression did not change with age in the medial habenula, medial geniculate nucleus or in the hippocampus whereas it decreased in the cortex. The developmental pattern of the hybridization signal for alpha 4 was different according to the brain area considered. The expression of the two transcripts showed a biphasic pattern in some thalamic nuclei: the lowest levels occurring during the first and second postnatal weeks respectively, and the highest levels during the second and fourth postnatal weeks. The ontogenetic profile of the expression of the alpha 4 subunit in the thalamic nuclei coincided with that of [3H]-L-nicotine binding sites. These findings suggest that the two subunits of nAChRs are independently regulated in most of the brain areas examined, and that in some regions, such as the thalamus, the ontogenetic variations reported for the alpha 4 subunit correlate with those observed for the [3H]-L-nicotine binding sites.

Aging↗

The genes encoding the biotin carboxyl carrier protein and biotin carboxylase subunits of Bacillus subtilis acetyl coenzyme A carboxylase, the first enzyme of fatty acid synthesis.

The genes encoding two subunits of acetyl coenzyme A carboxylase, biotin carboxyl carrier protein, and biotin carboxylase have been cloned from Bacillus subtilis. DNA sequencing and RNA blot hybridization studies indicated that the B. subtilis accB homolog which encodes biotin carboxyl carrier protein, is part of an operon that includes accC, the gene encoding the biotin carboxylase subunit of acetyl coenzyme A carboxylase.

Acetyl-CoA Carboxylase↗

Defective activity of Na+,K(+)-ATPase in peripheral nerve of diabetic rats is independent of the axonal transport of the enzyme.

This study addressed the question as to whether the reduced activity of Na+,K(+)-ATPase reported to occur in diabetic nerves and to play a crucial role in the pathogenesis of diabetic neuropathy could be due to derangements in the axonal transport of the enzyme. A micromethod was developed to evaluate the ATPase accumulation in individual segments of ligated sciatic nerves from streptozotocin-induced diabetic rats. The results confirmed a approximately 40% decrease in the background activity, but showed that the enzyme was transported at similar rates in both anterograde and retrograde directions, suggesting that the decrease in its activity does not depend on an altered delivery along the axons.

Analysis of Variance↗

Localization and age-dependent expression of hexokinase mRNA in the rat brain.

Localization of mRNA encoding for the enzyme hexokinase and its regulation in aged animals was carried out in rat brain using the in situ hybridization technique. The highest levels of the hybridization signal were observed in the olfactory bulb, piriform cortex, tenia tecta, hippocampus and granular cells of the cerebellum. Other brain areas and nuclei including cerebral cortex, thalamus, hypothalamus, substantia nigra, subiculum, choroid plexus and superior colliculus displayed moderate to low density of transcripts. Correlation between relative hexokinase content and levels of its mRNA was found only for some brain regions such as caudate-putamen, geniculate nucleus, ventral and lateral thalamic nuclei, superior colliculus and granular cells of the cerebellum. In the cerebral cortex and hippocampus of old animals the expression of hexokinase was significantly increased at 18 and 24 months of age. From the present data we conclude that although hexokinase is an ubiquitous enzyme, sites of synthesis display a discrete and uneven localization in rat CNS and expression, in the aging brain, might be regulated to compensate for reduced oxidative phosphorylation in the brain tissue.

Aging↗

[Postoperative ischemic ileocolitis in the elderly. Suggested therapy with intraluminal administration of oxygen and glutamine].

Ischaemic ileocolitis in postoperative course of major abdominal surgery is a great challenge for the surgeon: the mortality rate is very high, and therapeutic choices are poor. In the elderly patients ischaemic bleeding ileocolitis is often determined by low flow: sepsis and cytologic damage are primed by activation of endotoxins and chemical mediators, and bacterial translocation could develop across intestinal wall. In our case the patient (male, caucasian, 68 years old) underwent bilio-hepatic resection for hilar cholangiocarcinoma. In the postoperative period continuous enteric haemorrhage was determined by an ischaemic ileocolitis demonstrated by colonoscopy. Abdominal angiography did not show stenosis or occlusion of mesenteric vessels. We administered dopamine and dobutamine as vasodilator drugs for splanchnic circulation without any positive response. Surgical removal of the colon was unsuccessful to stop bleeding. ileostomy and sigmostomy were performed. Histologic samples of the specimen showed ischaemic ileocolitis. After a few days the patient bled again. As last therapeutic choice, we bubbled oxygen in a solution of L-glutamin 500 mml (3 liters/min for 5 min). We administered 500 mml of this solution three times a day by enteral sond, and 100 mml twice a day by sigmoidostomy and endoluminal oxygenation was performed twice a day (1l/min for 1-2 minutes) under continuous control. Bleeding was reducing during the next five days, until stopping. If glutamine and O2 can be considered the fuel of enterocytes, we hypothesized endoluminal oxygenation and glutamine enteral supply of the small intestine could feeding enterocytes, until a complete restoration of enteral mucosa and stopping of the haemorrhage.

Aged↗

Effects of postnatal or adult chronic acetylcholinesterase inhibition on muscarinic receptors, phosphoinositide turnover and m1 mRNA expression.

Muscarinic receptor number, receptor-stimulated phosphoinositide hydrolysis and m1 mRNA expression were examined in the cerebral cortex and hippocampus of rats treated during postnatal development or in adult age with the organophosphate diisopropylfluorophosphate. Developing rats were treated from postnatal days 4-9 or from postnatal days 4-20 and killed on days 10 and 21, respectively, 24 h after the last administration of diisopropylfluorophosphate. Adult animals were treated for 14 days. Acetylcholinesterase activity and muscarinic receptor number were significantly reduced in all groups of treatment. Muscarinic receptor-stimulated phosphoinositide turnover, however, was significantly reduced in postnatal days 4-20 and adult treated rats but not in the postnatal days 4-9 group. No differences were observed in ED50 values. Conversely, m1 mRNA expression was significantly reduced both in the cerebral cortex and hippocampus of postnatal days 4-9 treated rats, but not of postnatal days 4-20 and adult treated rats. These results indicate that chronic inhibition of acetylcholinesterase in developing rats results in significant alterations in muscarinic neurotransmission. These alterations may delay the maturation of the cholinergic system and, therefore, may account for some of the long-lasting neurotoxic effects observed after developmental exposure to organophosphate pesticides.

Acetylcholinesterase↗

Identification and functional significance of nicotinic cholinergic receptors in the rat pineal gland.

The existence and subunit identification of the nicotinic cholinergic receptors in the rat pineal gland were examined by autoradiography, using [3H]cytisine and [125I]alpha-bungarotoxin as labelled ligands. The experiments performed with radioactive cytisine did not reveal specific binding, while iodinated alpha-bungarotoxin disclosed moderate specific binding density, suggesting that the nicotinic cholinergic receptor in the rat pineal is structurally organized with the alpha 7 or alpha 8 subunits present, the only ones that bind alpha-bungarotoxin with high affinity. In vitro functional experiments using pineal explants demonstrated that the binding site may represent a readily accessible nicotinic cholinergic receptor. Nicotine, though having no effect per se on the synthesis and release of melatonin, significantly diminished, in a dose-dependent manner, the norepinephrine-stimulated melatonin accumulation. This effect could be blocked by coincubation with the cholinergic antagonist d-tubocurarine, suggesting that the nicotinic cholinergic receptor in the rat pineal could be involved in the functional regulation of the gland.

Alkaloids↗

Distribution of nicotinic receptors in cynomolgus monkey brain and ganglia: localization of alpha 3 subunit mRNA, alpha-bungarotoxin and nicotine binding sites.

The distribution of nicotinic receptors in the brain and ganglia of the Cynomolgus monkey was studied by in situ hybridization and receptor autoradiography. A 35S-labeled antisense riboprobe for the mRNA of the alpha 3 subunit of the human nicotinic receptor, [3H]L-nicotine and [125]alpha-bungarotoxin were used as markers. The highest levels of alpha 3-mRNA were observed in the hippocampus, the medial habenula, the lateral geniculate, the granular layer of the cerebellum, as well as in the pineal gland; moderate levels were found in other nuclei of the thalamus and in the deeper layers of the cerebral cortex. High-affinity binding sites for [3H]L-nicotine were observed mainly in the thalamus. The distribution of [125I]alpha-bungarotoxin binding sites was different from that observed for alpha 3-mRNA and [3H]L-nicotine; they were most abundant in a few specific thalamic nuclei, in the medial habenula and in lamina I of the cerebral cortex. The localization of these three markers was also investigated in the sympathetic, parasympathetic and sensory ganglia of the monkey. Intense labeling was observed for alpha 3-mRNA and for [125I]alpha-bungarotoxin in the sympathetic and parasympathetic ganglia, whereas no positive signal was seen in the ganglion of Gasser. [3H]L-nicotine binding was not detected in any of the ganglia examined. High levels of mRNA for the alpha 3 subunit of the nicotinic receptor were also detected in human sympathetic ganglia. Comparison between alpha 3-mRNA distribution and [3H]L-nicotine binding suggests that in the Cynomolgus monkey brain, the alpha 3 subunit may participate in the formation of more than one nicotinic receptor subtype: a high-affinity binding site for [3H]L-nicotine in the thalamus, and other sites with low affinity for nicotine in the medial habenula and cerebral cortex. Both the alpha 3-mRNA and the [125I]alpha-bungarotoxin are highly expressed in the sympathetic ganglia; however, since no information is presently available on the intraneuronal cellular localization, it cannot be established whether or not they are both present at synaptic sites.

Animals↗

Inner ester derivatives of gangliosides protect autonomic nerves of alloxan-diabetic rats against Na+, K(+)-ATPase activity defects.

Bovine brain gangliosides have been shown to prevent decay in Na+,K(+)-ATPase activity in sciatic and optic nerves of alloxan- and streptozotocin-diabetic rats. In the search for a drug with greater bioavailability and increased incorporation into neural tissue, ganglioside inner ester derivatives (AGF1) were recently developed. We evaluated the effect of AGF1 treatment on Na+,K(+)-ATPase activity in homogenates of vagus nerve from alloxan-diabetic rats (100 mg/kg s.c.). Animals were treated with AGF1: 10 mg/kg 6 days/week i.p., or 30 mg/kg biweekly i.p. Treatment began 10 d post-alloxan and continued for 8 consecutive weeks. Normal age- and sex-matched rats were used as controls. Alloxan intoxication produced a 39% decrease in Na+,K(+)-ATPase activity of the vagus nerve, which was completely restored (96-97% recovery) by both AGF1 regimes. Results suggest that ganglioside inner ester derivatives may be used in the clinical setting for the management of diabetic autonomic neuropathy.

Animals↗

Cytomegalovirus encephalitis in a non-immunocompromised patient: CSF diagnosis by in situ hybridization cells.

Cytomegalovirus (CMV) encephalitis in immunologically normal patients is rarely reported in the literature. CMV infection was diagnosed by viral DNA probe techniques on CSF cells in a 32-year-old, immunologically normal male presenting with a severe clinical picture due to encephalitis. Administration of ganciclovir was followed by an immediate improvement in the patient's condition. More sensitive techniques for CMV detection could allow to discover more cases of CMV encephalitis in non-immunocompromised patients than previously recognized.

Adult↗

Evaluation of Parkinson's disease: a new approach to disability.

Testing the reliability and usefulness of disability scales in Parkinson's disease has been the object of a study carried out by 4 neurologists on 48 patients using 2 rating scales--Hoehn and Yahr staging and Columbia University Rating Scale--and 2 disability scales--Northwestern University Disability Scale and Extensive Disability Scale, a new scale conceived for this purpose, which is more accurate in examining in a different way the physical incapacity and handicap of parkinsonian patients in their daily living. The examiners worked in pairs and did not discuss their rating scores before the end of the study. As far as interobserver agreement is concerned, the results of the disability scales are better than those of the rating scales. The Northwestern Disability Scale and the new scale are similar in reliability and consistency; however, the Extensive Disability Scale has proved to be a better indicator of the functional status. We suggest a more extensive employment of disability scales in clinical studies.

Activities of Daily Living↗