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Biomedical subjects

D Cavallotti

Publications and source records attributed to D Cavallotti.

At least 19 recordsLinked to original sources

Dopaminergic receptors in rat dura mater: pharmacological characteristics.

1. The location and distribution of dopaminergic receptors in rat dura mater was studied by examining several dural zones (vascular, perivascular, intervascular) in different cranial and spinal regions. 2. The pharmacological characteristics and anatomical distribution of dopamine D1- and D2-like receptors sites were investigated using combined pharmacological techniques and immunofluorescent microscopy. 3. Samples of rat dura mater were obtained from 10 adult Wistar rats. On frozen slices, dopaminergic D1 and D2 receptors were stained immunohistochemically using monoclonal antibodies. 4. Inhibition studies were performed using fluorescent and non-fluorescent agonists or antagonists to define the pharmacological specificity of the immunostaining. 5. The greater sensitivity to displacement by amisulpride, bromocryptine, domperidone, haloperidol, raclopride and l-sulpiride than to displacement by N-propyl-nor-apomorphine, quinpirole and clozapine suggests that the immunofluorescent sites observed in these experiments are likely to belong to the dopamine D2 receptor subtype. 6. Our observations provide evidence of the presence of D1 and D2 receptors in the wall of meningeal vessels. The dopaminergic receptors are located in the adventitia, media and intima of dural arteries. Furthermore, the density of receptors is higher in close proximity to arteries and decreases passing from the vascular to the perivascular and intervascular zones. 7. In the rat dura mater, dopamine regulates the meningeal blood vessels and, through this action, dopamine and its receptors can play an important role in the pathogenesis of cephalalgia.

Animals↗

Effect of interferon on human thymus microenvironment.

Interferon, a thymic immunostimulator, was used with the aim of assessing the importance of adrenergic nerve fibers (ANF) and/or AChE-positive nerve fibers (AChENF) in the regulation of some immunological functions in humans. Thymic normal control fragments and/or thymic fragments of immunostimulated patients were removed during surgical biopsies. Thymic slices were stained with eosin-orange (for the recognition of microanatomical details of the microenvironment) and with Bodian's method for staining of nerve fibers. Histofluorescence microscopy was employed for staining ANF. AChENFs were detected by means of the direct-coloring thiocholine method. All images were submitted to quantitative morphometrical analysis and statistical comparisons of data. Moreover, the amount of proteins and noradrenaline was measured on thymic homogenates of the same patients. Treatment with interferon induces substantial changes in the thymic microenvironment, on ANF, on AChENFs and on the total amount of proteins and noradrenaline in thymic tissue homogenates. In conclusion immunostimulation with interferon induces substantial changes in the whole thymus and in its microenvironment, involving both sympathetic and parasympathetic nerve fibers.

Adrenergic Agents↗

Age-related changes in rat optic nerve: morphological studies.

Age-related changes of the optic nerve were studied in 3-month-old (young), 12-month-old (adult) and 24-month-old (aged) male Sprague-Dawley rats. Cross sections of the intracranial portion of the optic nerves of animals of different age groups were stained with haematoxylin-eosin and examined under a light microscope at low and high magnification. Other sections were stained with crystal violet for demonstration of glial cells. A third group of sections were stained immunohistochemically to detect glial fibrillary acidic protein (GFAP) which is a marker for localizing and characterizing astrocytes. All morphological results were subjected to the quantitative analysis of images and to statistical analysis to identify significant morphometrical data. Tissue protein concentrations were determined on homogenized fragments of optic nerve. Our results demonstrate the following age-related changes: (1) increase of the optic nerve sheaths (meningeal membranes); (2) increased number of astrocytes; (3) increase of areal density of GFAP immunoreactivity; (4) increased diameter and area of the optic nerve; (5) decreased number of nerve fibres; (6) decreased-size of nerve fibres and (7) decrease of the nerve fibres/meningeal membrane ratio from 3:1 to 1:1. Moreover, the protein amount does not change with age. The rat optic nerve, therefore, appears sensitive to ageing processes and can be considered as a useful model for the studies on neuronal ageing.

Aging↗

Morphological changes in the sciatic nerve of diabetic rats treated with low molecular weight heparin OP 2123/parnaparin.

This study points out the early alterations that affect the sciatic nerve of rats with experimentally induced diabetes. It emphasizes the advantages deriving from treatment with low molecular weight heparin (LMWH) OP 2123/parnaparin and correlates these findings with observations emerging from a review of the relevant literature. In fact, the number and diameter of the capillaries within the sciatic nerve of diabetic rats appear to be increased consistent with a microangiopathy (the main characteristic of which is the fragility of new vessels), that is related to the evolution of diabetes in humans and animals. Our results suggest a possible therapeutic role for OP 2123/parnaparin in both diabetic neuropathy and microangiopathy, frequent complications of diabetes.

Animals↗

Interleukin 1beta as stimulator of the rat thymus.

The effects of interleukin 1beta administration on the thymus of adult and old rats were studied in order to study the interactions between the nervous and immune systems and to confirm the important role played by catecholaminergic nerve fibres (CNF) in the regulation of thymic functions. Moreover, chemical sympathectomy was performed in a group of rats to study the effects on thymus of the destruction of the majority of CNF. Our results indicate that thymic stimulation (performed by means of interleukin 1beta) induces substantial changes in the fresh weight of the whole thymus, as well as in the thymic microenvironment, thymic nerve fibres, CNF, neuropeptide Y (NPY)-like positive nerve fibres and total amount of both proteins and norepinephrine in rat thymic tissue homogenates. The majority of CNF are destroyed after chemical sympathectomy with 6-OH-Dopamine (DA) and remain unchanged after treatment with interleukin 1beta.

Adrenergic alpha-Agonists↗

Autosomal dominant distal myopathy: further evidence of a chromosome 14 locus.

In 1995 Laing et al. (Am J Hum Genet 56(1995)422) described a single family with nine members affected by an autosomal dominant infantile onset distal myopathy. This family generated a LOD score of 2.6 for a locus on chromosome 14. We describe two families with an infantile onset distal myopathy: a new family with four affected members and the family previously described by Scoppetta et al. (Acta Neurol Scand 92(1955)122) in both of which haplotype segregation was compatible with linkage to the same chromosome 14 locus, generating LOD scores of 0.9 at a penetrance of 100% for the markers D14S283 and D14S64 (theta=0) in both families. The loci for autosomal recessive hereditary inclusion body myopathy and Nonaka myopathy on chromosome 9 and for autosomal dominant distal myopathy of Markesberry-Griggs and Udd on chromosome 2q31-33 were excluded by linkage analysis. The disease followed a uniform course with selective wasting of the anterior tibial muscles, starting in infancy and recognizable by a characteristic clinical sign of the 'hanging big toe'. This was followed by slow progression, with involvement of the finger and wrist extensor muscles in the third decade and proximal limb muscles in the fourth decade. Interestingly, we also found evidence of an accompanying mild peripheral neuropathy in the oldest individual with hypomyelination of numerous large myelinated fibres. In addition, this patient's muscle biopsy also showed autophagic vacuoles and numerous intranuclear tubulo-filamentous inclusions of 15-20 nm diameter. Given that all three families with infantile onset distal myopathy are compatible with linkage to the same locus on chromosome 14, this study supports evidence for, and enlarges the clinical and neuropathological spectrum of the distal myopathy on chromosome 14.

Adolescent↗

Occurrence of catecholaminergic nerve fibers in the human uveoscleral tissue in conditions of normal and raised intraocular pressure.

The distribution of catecholaminergic nerve fibers (CNF) in human uveoscleral tissue was studied in six human eyes with normal intraocular pressure and in five eyes with increased pressure. The eyes with increased pressure had no visual field alterations and the patients did not have any glaucoma-related optic neuropathies. The amount of norepinephrine in these structures was also analysed. Catecholaminergic nerve fibers were detected by means of fluorescence microscopy and were counted using the quantitative analysis of images. Our results demonstrate that the occurrence of catecholaminergic nerve fibers (expressed in Conventional Units = C.U.) in human uveoscleral tissue is 15.4 +/- 1.6 C.U. in eyes with normal intraocular pressure. In eyes with increased intraocular pressure, these values were 12.2 +/- 1.2 C.U. Moreover, the amount of norepinephrine in tissue homogenates of the same eyes was evaluated and found to be 21.7 +/- 1.3 microg/gr tissue fresh weight of the human uveoscleral tissue in eyes with normal intraocular pressure. This value decreased to 18.8 +/- 1.1 microg/gr tissue fresh weight in the same tissue in conditions of raised intraocular pressure. In these experiments, the small number of eyes examined made it difficult to draw general conclusions. However, the role of human uveoscleral tissue was emphasized by the rich catecholaminergic innervation. A decrease of catecholaminergic nerve fibers and norepinephrine occurs when intraocular pressure is elevated. The modifications of these parameters, involved in the sympathetic control of aqueous humor outflow, may support the hypothesis of a possible relevant role for the human uveoscleral tissue in different pathological conditions.

Adult↗

Effect of interleukin 1beta on rat thymus microenvironment.

The effect of interleukin 1beta on the thymus of control and chemically sympathectomized adult and aged rats was studied with the aim of assessing the importance of adrenergic nerve fibres (ANF) in the regulation of some immunological functions. The whole thymus was removed from normal, sympathectomized (with the neurotoxin 6-OH-dopamine) and treated (interleukin 1beta) rats. Thymic slices were stained with eosin orange (for the recognition of microanatomical details of the thymic microenvironment) and with Bodian's method for staining of nerve fibres. Histofluorescence microscopy was employed for staining ANF and immunofluorescence was used for detecting NPY-like immunoreactivity. All images were submitted to quantitative morphometrical analysis and statistical analysis of data. Moreover, the amount of proteins and noradrenaline was measured on thymic homogenates. The results indicate that in normal conditions the formation of the thymic nerve plexi in the rat is complex: the majority of ANF are destroyed after chemical sympathectomy with 6-OH-dopamine and do not change after treatment with interleukin 1beta; on the contrary, treatment with interleukin 1beta induces substantial changes in the fresh weight of the thymus, the thymic microenvironment, thymic nerve fibers, ANF, NPY-like positive nerve fibres, and on the total amount of proteins and noradrenaline in rat thymic tissue homogenates. Immunostimulation with interleukin 1beta induces substantial changes in the whole thymus, in its microenvironment and in ANF and NPY-like nerve fibres. After chemical sympathectomy, no significant immune response was evoked by interleukin 1beta, since the majority of ANF was destroyed by chemical sympathectomy.

Adrenergic Agents↗

Interleukin 1beta and GABA-transaminase activity in rat thymus.

The occurrence and distribution of 4-aminobutyrate:2 oxoglutarate transaminase (GABA-t) activity were examined in the rat thymus of normal and immunostimulated rats using biochemical and histoenzymatical methods. Specific GABA-t reactivity was confined primarily to the arteries and, to a lesser extent, to the veins. Only a few activities could be observed in association with the subcapsular and medullar part of the parenchyma and nerve fibers. GABA-t was considered a linking enzyme between the immune and the nervous system and it was studied with the aim of analyzing the relationships between these two systems. Our findings indicate that the GABA-t activity in the thymus is specifically located in the wall of the blood vessels. Moreover, our results demonstrate the presence of a GABA-t activity in the peripheral blood vessels. Treatment with interleukin 1beta induces an increase of protein content of the amounts of GABA-t biochemically assayed and of the levels of histoenzymatically stained GABA-t. Furthermore, staining of the different structures of the thymus in treated or untreated rats shows that the significant modifications concern the parenchyma, the structures resembling nerve fibers and finally, the whole thymus. On the contrary, the highest activity of the GABA-t is located in the walls of arteries, veins and lymphatic vessels.

4-Aminobutyrate Transaminase↗

Quantification of acetylcholinesterase-positive structures in human thymus during development and aging.

Acetylcholinesterase (AChE) localization in the human thymus has been studied by biochemical and morphological methods during development and aging. The occurrence, the amount and the distribution of acetylcholinesterase and the changes with age were examined in 24 human thymuses. The whole human thymus was removed during autopsies in males of the following age-groups: prenatal of six months, new-born, infant, young, adult and elderly. The thymuses were weighed, measured and dissected: the microanatomical details were stained with Eosin-orange, nervous structures were identified by means of Bodian's method. Protein content was determined with biochemical methods. Histoenzymatical and biochemical demonstration of acetylcholinesterase was performed. The morphological results obtained were submitted to quantitative image analysis. Our results show that the thymic microenvironment changes with age; moreover, an increase of acetylcholinesterase-positive structures can be observed with age. Biochemical results are in agreement with morphological results and both are confirmed by the outcome of quantitative analysis of images. Acetylcholinesterase activity in human thymus may play a key role in thymic functions.

Acetylcholinesterase↗

gamma-aminobutyric acid-transaminase activity in the human thymus after administration of interferons.

The purpose of this article is to study the amounts of gamma-aminobutyric acid-transaminase (GABA-t) during immune response in the human thymus. GABA-t was studied by biochemical and histochemical methods in 7 immunostimulated (treated) and 7 non-immunostimulated (untreated) patients (4 young adult, age range: 24-36 years; 3 older adult, age range: 56-66 years). Immunostimulation was performed using interferon drugs for 6 months. After the histoenzymatic staining of GABA-t activity, the slides containing the samples of thymus of treated and untreated patients underwent quantitative analysis of images. The present results provide direct evidence that the immune response increases the level of GABA-t contained in vessels, parenchyma and nerve fibers of the thymus. Treatment with interferon is also capable of increasing the protein content of the thymus. The biochemical data together with the histoenzymatic results provide evidence for a localization of GABA-t in the thymic gland. Moreover, gamma-aminobutyric acid can be considered as one of the linking molecules between the immune and nervous functions of the human thymus.

4-Aminobutyrate Transaminase↗

Acetylcholinesterase activity in rat thymus after immunostimulation with interleukin beta.

The occurrence and distribution of Acetylcholinesterase (AChE) activity were examined in the thymus of normal and immuno-stimulated adult and aged rats using biochemical and enzymehistochemical methods. Specific AChE reactivity was found primarily in the arteries and, to a lesser extent, in the veins. Only a small amount of activity could be observed in association with the subcapsular and medullary part of the parenchyma and nerve fibers. Our findings indicate that AChE activity in the rat thymus increases after treatment with interleukin beta. In fact treatment with interleukin beta induces an increase of protein content, of the amounts of AChE biochemically assayed and at the levels of AChE histoenzymatically stained. Furthermore, staining of the different structures of the thymus in treated or untreated rats shows that the significant modifications concern the parenchyma, the structures resembling nerve fibers and the whole thymus, while only small changes are observed in AChE activity located in the walls of arteries, veins and lymphatic vessels.

Acetylcholinesterase↗

Microfilaments in regenerating cells of rabbit cornea: immunological and ultrastructural observations.

The role of myosin-like protein in regenerating and proliferating corneal cells following a standard alkali-injury in the rabbit eye has been studied. Microfilaments were observed by conventional transmission electron microscopy (T.E.M.) in injured epithelium and in fibroblasts. Typical microfilament bundles with electron dense structures and with stress fibers were evident. The presence of myosin-like proteins was demonstrated by means of immunochemical and autoradiographical techniques. Both epithelial cells and fibroblasts bind antimyosin-like antibodies (AMA). The same cells studied with the E.M. showed bundles of microfilaments in the cortical areas of their cytoplasm in correspondence with the same side of fluorescent or labelled AMA. The immunochemical and ultrastructural results suggest that both cells are able to produce in vivo movements involved in morphogenetic phenomena. Therefore, these structures play a role in post-traumatic corneal regeneration.

Actin Cytoskeleton↗

[Amnioinfusion: techniques, indications, and controlled retrospective study of 55 cases].

Amnioinfusion is a relatively recent procedure introduced among fetal medicine techniques. Its applications focus on two different methods: transcervical and transabdominal. The first procedure usually is carried out during "intrapartum amnioinfusion" to prevent or treat fetal heart rate (FHR) decelerations related to oligohydramnios or to dilute thick meconium staining of the amniotic fluid. The latter method used during "antepartum amnioinfusion" is usually indicated for severe oligohydramnios in order to avoid the complications related such as pulmonary hypoplasia, deforming effects of oligohydramnios, variable FHR decelerations and intraventricular hemorrhages. Antepartum amnioinfusion, also used to improve ultrasound visualisation in presence of oligohydramnios, is less employed as compared to intrapartum amnioinfusion, therefore its risks are not well established. In order to study possible adverse effects on the mother or foetus, fifty five patients affected by oligohydramnios at 17th-34th week of gestational age were submitted to antepartum amnioinfusion (1-5 procedures) and were matched retrospectively with forty seven women with the same characteristics treated with the conservative and expectant management. The trend of pregnancy was the same for both groups in relation to maternal fever > 38 degrees (10.9% in the amnioinfused group vs 17.0% in control group ns), leukocyte count > 18,000/mm3 (25.5% vs 21.3%, ns), C-reactive protein > 10 ng/ml (10.9% vs 6.4%, ns). The latency period between admission and delivery was significantly longer in the amnioinfused group than in the control one [21 (range 1-98) vs 9 days (range 0-72); p < 0.001] and the frequency of Apgar score < 7 at the 5th min was less represented in the amnioinfused group than in the control group (32.3% vs 66.6%; p < 0.001). In conclusion, it was interesting to note that antepartum amnioinfusion seems to increase the latency period between premature rupture of membranes and delivery, but it remains to clarify if this procedure is as much safe for the fetus as for the mother.

Adult↗

Occurrence of adrenergic nerve fibers and of noradrenaline in thymus gland of juvenile and aged rats.

Adrenergic nerve fibers (ANF) were studied in juvenile, adult and old rats by biochemical and morphological methods and by quantitative analysis of images (QAI). After chemical sympathectomy with neurotoxin 6-OH dopamine, the greater part of ANF disappeared. The whole thymus was drawn in juvenile normal or sympathectomized rats, in adult normal or sympathectomized rats and in old normal or sympathectomized rats. Thymuses from the above-mentioned animals were weighed, measured and dissected. Thymic slices were stained with eosin orange for the detection of the microanatomical details and with Bodian's method for the recognition of the whole nerve fibers. Histofluorescence microscopy was used for staining of ANF while immunofluorescence microscopy was employed for staining of neuropeptide Y (NPY)-like immunoreactivity. Biochemical dosage of proteins and of noradrenaline amount was performed. Finally, all morphological results were subjected to QAI. Our results suggest that: (1) total innervation of the thymus increases with age; (2) ANF do not change with age; (3) the content of noradrenaline in the thymus increases with age; and (4) NPY-like immunoreactive structures in the thymus decrease with age. Biochemical results are in accordance with the morphological ones and both are confirmed by means of QAI. The probable function of sympathetic innervation of rat thymus is also discussed.

Adrenergic Fibers↗

Occurrence of gamma-aminobutyric acid-transaminase activity in nerve fibers of human thymus.

The specific localization of gamma-aminobutyric acid-transaminase (GABA-t) in the thymus of young and elderly men was studied. Our results show a specific vascular localization of GABA-t in the human thymus, and deal with the amount and distribution of GABA-t and its changes with age. Samples of human thymus were harvested throughout of 12 autopsies in infants (n = 3), as well as young (n = 3), adult (n = 3) and elderly (n = 3) men. Histologic staining of the human thymus was performed with eosin-orange, while histologic staining of nerve fibers was performed with the Bodian method. Histochemical and biochemical demonstration of GABA-t, including protein dosage, was performed by the methods of Van Gelder and Jung, respectively. Finally, quantitative analysis of images was performed. Staining with eosin-orange reveals the micro-anatomical details of the thymic micro-environment. The Bodian method shows the nerve fibers and neurofibrils. Histochemical staining for GABA-t shows an increase of this enzyme with age and a marked localization in the nerve fibers of the thymus in infant, young, adult, and elderly men, as well as specific vascular localization of this enzyme. These biochemical data are in accordance with the histoenzymatic results and confirm all of our previous observations. Finally, quantitative analysis of images performed on slices let us confirm all the morphological changes induced by age. We can conclude that GABA is an inhibitory neurotransmitter of the human thymus, while GABA-t plays an important role in GABA metabolism.

4-Aminobutyrate Transaminase↗

Thymectomy and multiple sclerosis: ultrastructural study of an experimental model.

Transmission electron microscopy was performed on specimens of the thymus of rats induced for acute experimental allergic encephalomyelitis (EAE). The ultrastructural alterations of the thymus were progressive and correlated with EAE development. The thymic disorganization was due to a progressive degeneration of both epithelial cells and thymocytes. These data suggest a direct involvement of the epithelial thymic cells and thymocytes in EAE pathogenesis and may suggest the intriguing therapeutic concept of thymectomy in the management of multiple sclerosis.

Animals↗