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Biomedical subjects

G Di Trapani

Publications and source records attributed to G Di Trapani.

At least 37 records · Page 2Linked to original sources

Treatment of progressive supranuclear palsy with methysergide. A clinical study.

We present a clinical study of a patient aged 69 years with a clinical history of severe difficulty in walking and voluntary conjugate eye movement reduction in sideways glance and absence in vertical gaze. These symptoms led to diagnosis of Parkinson disease and treatment with L-Dopa + anticholinergics drugs. The treatment with antiparkinsonian drugs was suspended and no change in his clinical condition resulted. Methysergide therapy was initiated and the patient's response was monitored by video recording. On the basis of our experience and the data reported in the literature we believe that methysergide therapy affords some relief of symptoms in patients suffering from PSP.

Aged↗

Isolation from human placental extracts of a preparation possessing 'early pregnancy factor' activity and identification of the polypeptide components.

Protocols are described for the isolation from human placental extracts of a preparation which is active in the rosette inhibition assay by inducing an increased rosette inhibition titre. In this respect the preparation mimics the effects of pregnancy sera, an ability ascribed to the presence in these sera of a so-called 'early pregnancy factor' (EPF). In addition to this activity in the rosette inhibition assay, the preparation was also shown to modulate the expression of cell surface immunoglobulin on peripheral blood mononuclear cells. The polypeptide composition of the preparation was relatively simple, as revealed by SDS-PAGE. There was a major 12 kd polypeptide previously isolated in a rosette inhibition titre-active preparation from ovine placental extracts, and a small amount of 68 kd polypeptide. N-Terminal amino acid sequence analyses obtained after blotting onto polyvinylidene difluoride membranes identified the latter polypeptide as serum albumin and the major 12 kd polypeptide as human thioredoxin. Gel permeation analysis partially resolved activity expression away from the 12 kd polypeptide as activity expression was also found to be associated with low mol. wt material. It is concluded that EPF activity expression in pregnancy sera involves molecules related to and associated with thioredoxin.

Amino Acid Sequence↗

Clinical-electrophysiological correlations in a long-term case of subacute sclerosing panencephalitis with partial clinical improvement.

An electrophysiological follow-up was performed in a rare case of subacute sclerosing panencephalitis (SSPE) that from the vegetative stage had an extremely favourable course, lasting 4 years, in which clinical conditions passed, according to the Risk and Haddad scale, from stage 3 to stage 1. In this period of time, 5 CT scans and a series of EEGs, visual evoked potentials (VEPs) and brainstem auditory evoked potentials (BAEPs) were performed to investigate the correlation between clinical course and electrophysiological diagnostic data. The results showed a significant improvement in diagnostic results, directly correlated with the remission of clinical symptoms. Electrophysiological signs of improvement in VEPs were obtained before observable improvement was seen on CT scans. These results indicate that an accurate electrophysiological study (through cerebral evoked potentials) has an important role in investigating suspected cases of SSPE, in following the clinical evolution of this disease and providing prognostic data.

Adolescent↗

Congenital hydrocephalus: a new experimental model with histopathological study.

We describe a new experimental model of fetal hydrocephalus in the lamb. 14 sheep were operated on at 100-120 days gestation for the insertion of a catheter into the fetal aqueduct of Sylvius to block cerebrospinal fluid (CSF) flow. After the operation the intracranial pressure (ICP) was measured daily from the distal end of the catheter. The progress of ventricular dilatation was recorded by ultrasound. At ICP 100 mm/H2O the animals were killed for postmortem examination of the fetuses. Neuropathological examination showed massive dilatation of the ventricles. The ependymal cells appeared to be flat and the cellular lining disrupted. Growth of pseudocysts, cellular stratification and proliferation of the paraventricular germinal cells were observed also. With our new experimental model we were able to control the rise in ICP and correlate the evolution of the anatomical damage with the duration of high ICP and with the gestational age at which it began. Our model can also be used at early stages of gestation for reversing the development of hydrocephalus. It might therefore provide information on the suitability of fetal hydrocephalus surgery.

Animals↗

Modulation of rat peritoneal mast cell and human basophil histamine release by estrogens.

This study was undertaken to investigate the effect of estrogens on the histamine release mediated by IgE in rat peritoneal mast cells (PMC) and in sensitized human basophils. The estrogens were found to enhance the histamine release of either rat PMC and sensitized human basophils upon stimulation with anti-IgE. The enhancement was estrogens dose-dependent reaching the maximum value of 23% for rat PMC and 41% for sensitized human basophils stimulated with anti-IgE upon preincubation with 10(-8) M estrogens. Moreover, when purified PMC were used, the enhancing effect was still detected, suggesting a direct interaction between estrogens and mast cells. The enhancing effect took place quite rapidly reaching plateau levels in about 60 min. Basophils preincubated at 4 instead of 37 degrees C did not give any appreciable enhancement, suggesting that it was temperature-dependent and that the effect observed was not due to cytotoxicity. Incubation of PMC or human basophils with estrogens alone, without challenge with anti-IgE, did not give any detectable histamine release. The enhancement of histamine release by estrogens is probably mediated by IgE molecules present on the cell membrane, since this effect was not observed on challenge with substance P or compound 48/80, two segretagogues known to induce histamine release not via IgE.

Animals↗

Experimental allergic encephalomyelitis in guinea pig: variability of response to intradermal emulsion injection.

Different forms of experimental allergic encephalomyelitis were obtained in 4 groups of guinea pigs: 7 adult Hartley guinea pigs (Group I), 12 adults of the same strain (Group II), 6 juvenile strain 2 guinea pigs (Group III) and 6 juvenile strain 13 animals (Group IV), by the injection of emulsions. Groups I and II received emulsions containing 250 mg and 500 mg respectively of fresh isologous spinal cord tissue, complete Freund adjuvant (CFA) and saline solution while Groups III and IV received an emulsion containing 120 mg of isologous spinal cord, CFA, saline solution and 15 mg of Mycobacterium tuberculosis. The increased antigen load induced a disease with delayed onset and prolonged progressive course (C-P-EAE) in Groups I and II, although 8 animals showed no symptoms of illness. The findings in C-P-EAE were large demyelinated plaques, perivenous fibrosis and large areas of infiltration. Demyelinated areas occurred within the spinal cord white matter only in two asymptomatic animals. C-P-EAE was obtained in 4 of the Strain 2 animals. In conclusion, the increased antigen load induced a range of lesions in Hartley guinea pigs, although not all animals were affected. C-P-EAE was induced also in strains of guinea pig other than the Heartley strain. These different reactions may have been the outcome of partial or complete inactivation of the cell-mediated response to the inoculated antigens.

Acute Disease↗

Evoked potentials suggest cranial nerves and CNS involvement in chronic relapsing polyradiculoneuropathy.

Brainstem auditory and visual potentials were recorded from 6 patients with chronic idiopathic polyradiculoneuropathy. Evoked potentials were either completely absent or their latency was prolonged suggesting slow conduction in visual or auditory pathways. The results of our study provide further evidence suggesting subclinical involvement of cranial nerves and central nervous system sensory pathways in chronic inflammatory demyelinating polyradiculoneuropathy.

Adult↗

Identification of a histamine-releasing factor secreted by human pre-implantation embryos grown in vitro.

Human pre-implantation stage embryos cultured in vitro spontaneously secreted a factor capable of inducing histamine-release from human blood basophils. The embryo-derived histamine-releasing factor (EHRF) has been isolated from the culture medium by means of heparin-Sepharose affinity chromatography. The factor bound to the column and was then eluted by increasing the buffer molarity to 1.5 M NaCl. EHRF was detected using an enzymatic-isotopic microassay and sensitized basophils known to undergo release with anti-IgE. The EHRF-induced histamine-release was calcium and temperature dependent and the relatively slow kinetics (10 min) were similar to those obtained with anti-IgE. EHRF caused the release of a substantial amount of histamine (48%, n = 18) in a dose-dependent manner. The equivalent fraction isolated from medium containing unfertilized oocytes gave less than 10% of histamine-release using the same source of basophils, suggesting that EHRF was secreted after fertilization. EHRF was very stable since it was resistant to boiling, lyophilization, and to several freeze and thaw treatments. The histamine-releasing activity induced by EHRF was measured in vitro also by means of purified leukocytes containing sensitized basophils. EHRF could represent a message sent by the embryo to the mother to induce histamine release at the time of implantation.

Antibodies, Anti-Idiotypic↗

Dispersal of rat uterine mast cells and their functional response to an embryo-derived histamine releasing factor: a possible model for embryo implantation.

Rat uterine tissue was dissociated by enzymatic digestion with collagenase and viable mast cells were obtained. Their viability was assessed by the ability to exclude trypan blue dye and to respond functionally to different stimuli. Challenge with anti-IgE gave a calcium-dependent histamine release of 49%, whilst the undigested uterine fragments gave 23%. Moreover, they were capable of releasing histamine on challenge with the compound 48/80, suggesting a similarity with connective tissue mast cells. This similarity was further supported by their insensitivity to aldehyde blocking of dye binding. The final dispersed cell preparation contained 3 X 10(5) mast cells/g of uterine tissue, representing about 2% of total nucleated cells. The total histamine content of the undigested uterus was 2.5 micrograms/g of tissue, whilst after digestion the histamine determined was 1.2 pg per mast cell with a yield of 14%. The total histamine content of the uterus changed throughout the reproductive cycle, increasing before ovulation, reaching a maximum during ovulation and then decreasing after embryo implantation. This suggests that the implanting embryo, interacting with the uterus, may be capable of inducing the release of histamine. The embryo-derived histamine releasing factor (EHRF) that we have described previously is capable of inducing 22% histamine-release on uterine mast cells, thus supporting this hypothesis.

Animals↗

Nerve fibres in osteoid osteoma.

Many authors have attempted to explain the mechanism of pain in osteoid osteoma and, in the past, several histological studies have been carried out with the specific aim of verifying the existence of nerve fibres within the tumour. The histochemical procedures based on the affinity of nerves for silver stains have made it possible to demonstrate the presence of fine nerve fibres disposed alongside and around the vessels. An analogous study was conducted to confirm this and to supply further morphological and functional information useful for an understanding of the genesis of pain. The study involved three cases of osteoid osteoma using both traditional histochemical procedures and electron microscopy. The latter, in particular, allowed us to confirm the myelinic nature of the nerve fibres observed under optic microscopy. Based on these observations, the authors present a hypothesis on the function of the nerve fibres in the mechanism of pain stimulation.

Bone Neoplasms↗

Light and ultrastructural studies in sural biopsies of the pseudopolyneuropathic form of ALS.

The aim of this work was to study the neuropathological picture of the sural nerve in the pseudopolineuropathic form of A.L.S. Five patients were considered: in all cases the clinical and electromyographic follow-up excluded other diseases. EMG-studies were repeatedly performed: they showed the progressive evolution of the spinal anterior horn cell pathology from lower spinal to cervical levels. The sural nerve was processed according to the procedure used in our laboratory for light and ultrastructural examination. Quantitative analysis of myelinated fiber density was carried out on photographic enlargement and reported on histograms. The light and ultrastructural examination showed a severe loss of myelinated fibers; the decrease affected all types of fibers, but predominated in the largest ones. In the axons there was an increase of mitochondria, dilation of the small vesicles and increase in the number of neurofilaments. It is our opinion that all histopathologic data obtained in the sural nerve biopsy in this form of A.L.S. reveal a clear involvement of the sensory neurons of the spinal ganglia and the results can be useful for the study of precocius lesions in the A.L.S.

Amyotrophic Lateral Sclerosis↗

Isolation of a histamine releasing factor from human embryo culture medium after in-vitro fertilization.

In-vitro fertilization (IVF) of human oocytes in our laboratories gave a percentage pregnancy rate per transfer close to 20% during 1985. Embryos were grown until the two-four cell stage and then transferred to the maternal uterus. The media from these embryo cultures were collected and subjected to chromatography on heparin-Sepharose affinity columns. The bound protein fraction contained a factor capable of inducing histamine release from sensitized basophils. The effect of this embryo-derived histamine-releasing factor (EHRF) was to induce a maximum 56 +/- 7% release of the total histamine available. This value varied between 20 and 60%, resulting from 10-30 micrograms/ml of EHRF. Since the histamine release assay performed with basophils from non-atopic donors gave no positive results, we conclude that the release was not due to a cytotoxic mechanism. This was also supported by the absence of histamine release when the assay was performed at 0 degree C, or in the presence of 2 mM EDTA, suggesting that release was dependent on an immunological interaction between EHRF and some receptor on the basophils. The immunosuppressive role of histamine is well known, and a model involving EHRF and histamine is suggested here to explain the mechanism mounted by the embryo to escape maternal immune rejection.

Culture Media↗

Peripheral neuropathy in course of progressive systemic sclerosis. Light and ultrastructural study.

Progressive systemic sclerosis (PSS) is a chronic inflammatory disease of the connective tissue with involvement of the skin and other organs. The disease is characterized by an abnormal accumulation of collagen in all tissues and by microangiopathy. The involvement of the peripheral nervous system during PSS is very unusual and few cases are reported in the literature. A morphological study on the neuropathy associated with sclerodermia has been performed in rare cases. In this paper we demonstrate the role that the vascular lesions have in the pathogenesis of neuropathy during scleroderma. In particular, the primary role of the peripheral microangiopathy during PSS (observed in different clinical cases) is verified.

Female↗

Dementia-peripheral neuropathy during combined deficiency of vitamin B12 and folate. Light microscopy and ultrastructural study of sural nerve.

We present a case of a 56 year old woman who, in consequence of repeated surgical resections of the small bowel, developed a slowly progressive and mainly motor polyneuropathy affecting the lower limbs. The polyneuropathy due to a combined deficiency of vitamin B12 and folate, was associated with intellectual deterioration, involving especially the mnesic functions. Light microscopy and ultrastructural study of sural nerve revealed: marked reduction of every order of myelinated fibers: aspects of wallerian-like myelin degeneration; various stages of myelin leaflet delamination into the cytoplasm of Schwann cells and/or macrophages; lack of alterations attributable to a primary axonal degeneration. The morphological results suggest a primary demyelination, followed secondarily by either axonal changes or some aspects of wallerian-like degeneration.

Brain↗

Immunosuppressive effect of early pregnancy factor on early expression of cell surface membrane IgG.

We have adopted a new assay to investigate the influence of early pregnancy factor (EPF) on the modulation of lymphocyte activity. Lymphocytes were attached to the plastic surfaces of microplates in serum-free medium in the presence of Sepharose-Con A. After 2-3 days incubation with EPF, and ELISA assay was used to detect the expression of surface membrane IgG (smIgG); this was done in the same microplates used for the culture, thus avoiding cell manipulation. Using only a few picograms of EPF a significant inhibition (in the range 26-40%) was obtained. The variation in the inhibition observed was mainly due to the different sources of lymphocytes used. Unrelated proteins and hormones, tested at the same concentration as EPF, did not show any inhibitory activity. Using the F(ab)2 fragment of anti-human IgG instead of the whole molecule the same levels of inhibition were obtained, suggesting that the observed inhibition by EPF was not due to a non-specific interaction between the anti-human IgG and the Fc receptors on the cell. Such inhibitory activity detected in vitro by this method provides additional support for a suppressive role for EPF during pregnancy.

Adult↗

A new method to detect directly in culture cell surface membrane immunoglobulins.

An ELISA assay is described for the measurement of the smIgG. The method is based on the detection of cell-smIgG directly on the same microplate used for the culture. The cells, preincubated at 37 degrees C for one hour, were cultured in the presence of S-ConA and serum-free medium for two days. Using this strategy, the background noise due to non specific adsorption of IgG to plastic wells and cytophilic antibodies was eliminated. The cells in the presence of S-ConA and serum-free medium adhered to the plastic wells, and the cell-smIgG were detected using an anti-human IgG covalently linked to alkaline phosphatase or its F(ab')2 fragment. The possibility of measuring the modulation of the expression of the cell-smIgG without any additional manipulation is stressed.

Adult↗

Binding of human EPF to receptors on PMBC as a first signal of pregnancy immunomodulation.

The interaction between human early pregnancy factor (EPF) and a specific receptor present on human peripheral blood mononuclear cells (PMBC) has been assessed. EPF was radioiodiated by the lactoperoxidase method to a high specific activity, and the [125I]EPF obtained bound in a specific and saturable manner to the receptors on PBMC. Saturation of the binding sites occurred at 10(-9) M. There were 6600 specific binding sites per cell in cells partially purified from pregnant women and fewer in those from non-pregnant women. Unlabelled EPF was capable of competing for binding sites with [125I]EPF, while the binding of [125I]EPF was not inhibited by human chorionic gonadotrophin. A new assay was used that permits the culture of PBMC and the detection of the surface IgG expression in the same microplate. With this method the influence in vitro of human EPF on lymphocyte expression was tested. The results demonstrated a specific inhibition of surface IgG expression of PBMC using a very low concentration of EPF (10 pg/ml).

Binding, Competitive↗