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Biomedical subjects

P Bruni

Publications and source records attributed to P Bruni.

At least 55 records · Page 3Linked to original sources

Bradykinin increases ceramide and sphingosine content in human fibroblasts: possible involvement of glycosphingolipids.

Sphingolipid-derived products are recognized as second messengers able to mediate the action of extracellular signals such as cytokines and growth factors. In the present study it is shown that also bradykinin (BK), a pro-inflammatory peptide acting through G-protein-coupled receptors, is able to activate sphingolipid metabolism. Fibroblast treatment with the peptide provokes a rapid and significant increase in ceramide followed by a transient rise in sphingosine content. Sphingomyelin does not appear to be the source of ceramide since BK was unable to decrease the [3H]sphingomyelin pool and no increase in neutral or acidic sphingomyelinase activities could be detected after treatment with the peptide. The observation that the labeled glycosphingolipid pool is decreased upon BK stimulation would rather suggest that the peptide increases ceramide cellular content by rapidly mobilizing neutral glycolipids. Even though the physiological relevance of ceramide and sphingosine increase induced by BK is not known, it is noteworthy that glycosphingolipids may participate in this lipid signalling pathway.

Bradykinin↗

Fructose 2,6-bisphosphate metabolism during megakaryocytic differentiation of K562 and MEG-01 cells.

Fructose 2,6-bisphosphate (Fru-2, 6-P2) represents the most powerful activator of 6-phosphofructo-1-kinase, rate-limiting enzyme of glycolysis. Fru-2,6-P2 content is tightly regulated and appears to be under the control of different hormones and growth factors, acting either through covalent modification of isoenzymatic forms of 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase (PFK-2/FBPase-2), the bifunctional enzyme responsible for the synthesis and the degradation of the compound, or through changes in transcription rate and/or in the expression of different isoforms of the enzyme. In the present study the metabolism of Fru-2,6-P2 was investigated during the differentiation toward megakaryocytes induced by phorbol 12-myristate 13-acetate (PMA) treatment of human leukemia K562 and MEG-01 cell lines. Fru-2,6-P2 content as well as PFK-2 activity were increased in a dose-dependent manner after 4 days of incubation with PMA. MEG-01 cells resulted more sensitive to the effect of the inducer, anyway in both cell types cytostatic concentrations of the phorbol ester were able to affect Fru-2,6-P2 metabolism. The effect of PMA was maximal at 4 days of incubation in both examined cell lines. Interestingly, the effect induced by the phorbol ester at 4 days was still appreciable subculturing K562 and MEG-01 cells for 3 days in the absence of the inducer and was associated with relevant changes in the molecular properties of PFK-2: namely increased Vmax and K(m). This latter finding suggests that the rise in Fru-2,6-P2 content during the differentiation process toward megakaryocytes might result from the expression of a novel PFK-2 isoform.

Carcinogens↗

Life threatening intracranial complications of frontal sinus osteomas: report of two cases.

Paranasal sinuses osteomas are known as biological benign tumours. However, due to the peculiar anatomical relationships, patients harbouring an osteoma within the frontal sinus are exposed to serious orbital and intracranial complications. We report two unusual cases of intracranial mucocoeles associated with frontal osteomas. In one of them, a superposed tension aerocoele required emergency surgery. Although aggressive treatment of asymptomatic osteomas is not warranted, these lesions must be carefully observed and resected as soon as they show clinical and/or radiological signs of progression. The physiopathological and clinical aspects are discussed.

Bone Neoplasms↗

Surgical treatment of spasmodic torticollis: is there a role for microvascular decompression? With an illustrative case report.

Spasmodic torticollis (ST) is a puzzling movement disorder, characterized by involuntary tonic or clonic contractions of various neck muscles. From time to time, psychogenic, extrapyramidal and neuroperipheral origins have been postulated and reflecting the variety of theories proposed, as many different treatments have been attempted, none of which has shown absolute effectiveness. Surgery of ST classically includes destructive procedures such as myotomies, stereotactically placed lesions, rhizotomies and neurectomies. The recent application of the concept of "neurovascular conflict" to ST induced several authors to perform microvascular decompression (MVD) in these patients, with encouraging and in some cases long-lasting results. Our case report joins this group. From the analysis of pertinent literature we conclude that: a. Spasmodic torticollis is probably a collection of separate clinical entities; b. Even in the so-called "ST of neuroperipheral origin" some coexising central factor must be admitted; c. Neurovascular compression underlies at least some cases of ST. Historical, anatomical, physiopathological and clinical aspects of ST are extensively discussed.

Decompression, Surgical↗

Agonist-regulated L-arginine uptake in human platelets. Evidence against intracellular utilization of the amino acid.

L-arginine uptake takes place in human platelets through a saturable high affinity Na(+)-independent process mediated by the y(+) carrier for cationic amino acids. In the present study the effect of thrombin and collagen on L-arginine transport in human platelets was investigated. Thrombin significantly affected L-arginine uptake whereas collagen was uneffective. In particular, thrombin increased Vmax of the uptake by 77%, while it reduced the affinity of the carrier for L-arginine. The effect of thrombin on the transport process did not result in any increase in L-arginine metabolism since no conversion of the amino acid, either to L-citrulline (indicative of the presence of the L-arginine/nitric oxide pathway) or to any other metabolite, could be detected in resting or stimulated platelets.

Arginine↗

1,25-dihydroxyvitamin D3 inhibits proliferation of IMR-90 human fibroblasts and stimulates pyruvate kinase activity in confluent-phase cells.

This study tested the hypothesis that 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) plays a role in regulating some aspects of metabolism in IMR-90 normal human fetal lung fibroblasts. Among the enzymes studied, only pyruvate kinase showed a significant increase after treatment of confluent-phase cells with 1,25(OH)2D3 at various concentrations (0.1-100 nM range) for 24 h. A parallel increase in lactate output was observed. Steroid specificity was established by the failure of 10 nM levels of 25-hydroxyvitamin D3, estradiol-17 beta and progesterone to affect pyruvate kinase activity. The determination of the time course of [3H]-2-deoxy-D-glucose transport indicated that the hormone did not influence the transmembrane transport system of D-glucose. The addition of the inhibitors cycloheximide and actinomycin D to the culture medium abolished, at least in part, the 1,25(OH)2D3 stimulation of pyruvate kinase activity, suggesting the probable dependence of the hormone effect on cellular RNA and protein synthesis. 1,25(OH)2D3 also affected fibroblast growth and DNA synthesis. Cell number significantly decreased after 2-5 days treatment with 10 nM hormone in comparison with control fibroblasts, and also the incorporation of [3H]thymidine into DNA decreased after treatment of the cells with 1 and 10 nM hormone for 48 h. In conclusion, these data demonstrate that 1,25(OH)2D3 stimulates pyruvate kinase activity in confluent-phase IMR-90 human fibroblasts by a mechanism probably dependent on de novo protein synthesis, and also affects cell growth and DNA synthesis in sub-confluent-phase cells.

Biological Transport↗

Potentiated bradykinin-induced increase of 1,2-diacylglycerol generation and phospholipase D activity in human senescent fibroblasts.

1. The comparative study of the effect of bradykinin (BK) in young and old IMR-90 human fibroblasts shows that old cells are characterized by a reduced increase in 1,2-diacylglycerol (1,2-DAG) generation upon stimulation after short-term treatment and a significant higher increase after long-term agonist treatment. BK-induced activation of phospholipase D (PLD), the major enzyme involved in sustained 1,2-DAG generation, was 2.5-fold higher in old cells, strongly suggesting that it is involved in the potentiated increase of 1,2-DAG formation. The increased activation of PLD by BK in old cells was specific, since in parallel experiments the effect of thrombin was not significantly different in young and old cells. PLD activity in old cells was only reduced by down-regulation of protein kinase C (PKC) activity, in contrast to what was observed in young cells where it was completely abolished. This indicates that the enzyme activity in old cells was partially PKC-independent. BK was also able to increase the release of [14C]ethanolamine, a water-soluble product of hydrolysis of phosphatidylethanolamine (PtdEtn), through PLD activation in young and old cells. The BK effect was significantly higher in old cells and, very likely, PKC-independent, since phorbol 12-myristate 13-acetate failed to induce PtdEtn hydrolysis. 2. The present results indicate that the PLD/1,2-DAG pathway is specifically potentiated by BK in old fibroblasts, demonstrating that the formation of positive effectors of PKC activation is not necessarily decreased in cellular senescence. It remains to be established whether the increased generation of DAG upon BK stimulation plays any role in the altered PKC signalling pathway which characterizes old fibroblasts.

Bradykinin↗

Identification of a specific transport system for L-arginine in human platelets.

The present study demonstrates that human platelets possess a specific L-arginine transport system able to provide adequate amounts of L-arginine for endogenous nitric oxide production. L-arginine uptake takes place through a saturable high affinity carrier-mediated Na(+)-independent process which is significantly inhibited by L-ornithine, L-lysine and N omega-methyl-L-arginine. The high affinity of the transport process and the pattern of inhibition are consistent with mediation of L-arginine transport via the Na(+)-independent y+ system. The data on the kinetic parameters of the transporter suggest a possible role for arginine plasma levels in the regulation of platelet nitric oxide production.

Arginine↗

Glutamine transport and enzymatic activities involved in glutaminolysis in human platelets.

Glutamine is actively metabolized in human platelets, representing a preferential mitochondrial oxidative substrate in these cells. The primary importance of this metabolic route of glutamine is further confirmed here by the observation that platelet glutaminase activity is entirely represented by the phosphate dependent glutaminase or glutaminase I, most probably localized in the mitochondrial platelet fraction and classified by kinetic analysis as a kidney-type form. The following step of the glutamine metabolizing pathway, allowing the entrance of the amino acid skeleton carbons in the Krebs cycle, might be catalyzed by both glutamate dehydrogenase and aspartate transaminase, the first being entirely mitochondrial and the latter 65% mitochondrial. We also investigated platelets for the presence of one or more specific transport systems involved in glutamine uptake and we present the first evidence for two glutamine transport systems in human platelets that by inhibition analysis appear to share characteristics with the Na(+)-dependent ASC system and the Na(+)-independent L system for dipolar amino acid uptake. Both systems display affinity characteristics for glutamine in the range of plasma glutamine concentration and may thus have physiological relevance for the uptake of the amino acid in these cells.

Alanine↗

Acromegaly and pituitary tumors: early anatomoclinical observations.

The elucidation of pituitary physiopathology has been a major challenge for physicians since early ages. Due to the unawareness of the endocrine system, acromegaly, the most striking pituitary disorder, was commonly regarded as an intrinsic bone disease. Andrea Verga and Vincenzo Brigidi, Italian authors of the 19th century, reported the first macroscopic and microscopic descriptions of pituitary adenomas in acromegalic patients. Although far from providing a correct pathogenetic interpretation of the disease, they opened the way to forthcoming observations and discoveries. A short history of acromegaly and pituitary physiopathology is drawn.

Acromegaly↗

Meat allergy: I--Specific IgE to BSA and OSA in atopic, beef sensitive children.

OBJECTIVE: The use of lamb meat products has been suggested as an alternative diet for polyallergic children, although until now this clinical practice has not been supported by in-depth biochemical/immunological studies. The aims of this research were: to evaluate cross-reactivity between lamb and beef; to evaluate the role of BSA and OSA as allergens in beef allergic children; and to evaluate cross-reactivity between BSA and OSA. METHODS: 16 children suffering from atopic dermatitis (AD), aged 12 months-8 8 years (mean age 2.61 +/- 1.93 years) were found skin prick test (SPT)--positive to bovine meat; all of them were also SPT-positive to ovine meat and to milk. After a period of restricted diet, the selected 16 children were recalled; 12 AD-free children (8 males and 14 females, aged 12 months-4.33 years (mean age 2.21 +/- 1.05 years) were evaluated by SPT and radioallergosorbent test (RAST) for the following allergens: bovine meat, ovine meat, BSA 1 mg/ml, OSA 1 mg/ml. Double-blind, placebo-controlled food challenge (DBPCFC) with bovine serum albumin (BSA) and ovine serum albumin (OSA) were performed. For SPT, the results were expressed in mm of wheal, and 3 mm was considered as the end point; correlation between wheal diameters was calculated by Spearman rank test. For DBPCFC, according to the Sampson's experimental procedure, BSA and OSA were given in pear juice (the dermal negative response to the pear juice was verified by fresh food SPT before starting the oral challenge test). The total dose administered to the children corresponded to the amount of albumin present in 180 g of calf or lamb meat (90 and 63 mg respectively, as calculated by Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SPS-PAGE). The administration of pear juice (containing placebo or albumin) and symptom evaluation were entrusted to medical people who did not know what the child received. RESULTS: All children tested SPT positive to bovine and ovine meat, and to BSA and OSA. Significant correlations were observed between the following diameters of wheal: BSA vs OSA (R = 0.846, p < 0.0001); ovine meat vs OSA (R = 0.769, p < 0.005); b.meat vs o.meat (R = 0.771, p < 0.005); and ovine meat vs BSA (R = 0.594, p < 0.043). In RAST, 6 of 12 children were positive to bovine meat, 3 to lamb meat, 4 to BSA and 3 to OSA. DBPCFC showed an immediate reaction to BSA or OSA in 2 and 3 children, respectively. One other child developed severe dyspnea, cough and asthma 3 hours after OSA challenge. CONCLUSIONS: BSA and OSA are important beef and lamb allergens; they share not only proteic sequences, but also allergenic properties. Clinical tolerance to BSA and OSA can be present in beef and lamb SPT-positive children.

Animals↗

Celebrating the centennial (1894-1994): Leonardo Gigli and his wire saw.

In spite of the recent introduction of craniotomes in neurosurgical practice, the simple but brilliant wire saw invented by Leonardo Gigli still holds an important place in neurosurgical instrumentation. Born in Florence in 1863, Gigli was forced by circumstances to leave Italy soon after getting his medical degree. He first spent 2 years attending the most celebrated obstetrical clinics in Paris and London and then, in 1892, moved to Breslau where he worked with Fritsch and Mikulicz. During this successful and rewarding period, Gigli proposed the lateralized pubiotomy (Gigli's operation) for safe delivery in cases of maternal pelvic deformities and, inspired by the sight of a jagged knife during a country banquet, conceived his wire saw to simplify the procedure. In 1894, at Professor Obalinski's suggestion, he successfully tested a modified saw type with a whalebone guide for the preparation of osteoplastic cranial flaps. In spite of his great popularity and the high esteem in which he was held abroad, Gigli's aims were systematically belittled in Italy, where he never qualified for a university teaching position. He died in 1908, at age 44. Although the once celebrated Gigli's operation has merely historical interest today, the favorable features of his wire saw make it a safe and efficient tool in the hands of twentieth-century neurosurgeons worldwide.

History, 19th Century↗

Involvement of protein kinase C and arachidonate signaling pathways in the alteration of proliferative response of senescent IMR-90 human fibroblasts.

The proliferative response of IMR-90 fibroblasts at low and high population doubling level (PDL) to protein kinase C activation has been investigated to clarify whether the reduced mitogenic responsiveness of senescent cells can be ascribed to an alteration in protein kinase C signal transduction pathway. The results show that the signaling pathway leading to DNA synthesis through protein kinase C activation, appears to be modified in senescent IMR-90 human fibroblasts. High PDL fibroblasts exhibit a different sensitivity to phorbol 12-myristate 13-acetate (PMA) and dioctanoylglycerol (diC8); high glucose reduced responsiveness to PMA only in these cells. In addition, high PDL fibroblasts are characterized by an increase in diacylglycerol (DAG) cellular mass that could contribute to the different regulatory properties of the signaling pathway. On the other hand, the ability of the cyclooxygenase inhibitor indomethacin to strikingly improve the proliferative response of high PDL cells to PMA indicates that an altered overall metabolism of arachidonate may represent a crucial step in the reduced mitogenic response involving protein kinase C activation.

Arachidonic Acid↗

Bradykinin stimulates fructose 2,6-bisphosphate metabolism in human fibroblasts.

Bradykinin (BK), a peptide released during inflammatory response, has been investigated for its ability to regulate glucose metabolism in human fibroblasts. The peptide is able to significantly increase glycolytic flux in these cells. The strict relationship between the glycolytic rate and the levels of fructose 2,6-bisphosphate (Fru-2,6-P2) strongly suggests that the metabolite plays a key role in the regulation of glucose metabolism by bradykinin. The mechanism by which bradykinin increases Fru-2,6-P2 content involves the activation of 6-phosphofructo-2-kinase (PFK-2), the enzyme responsible for the synthesis of the metabolite. The study of the multiple signalling systems triggered by bradykinin demonstrates the involvement of the rise in intracellular Ca2+ concentration and of protein kinase C mediated pathway in the mechanism by which bradykinin increases Fru-2,6-P2 content and PFK-2 activity.

Bradykinin↗