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Biomedical subjects

A Castelli

Publications and source records attributed to A Castelli.

At least 55 records · Page 3Linked to original sources

Mixed function oxidation and enzymes: kinetic and structural properties of an oxidatively modified alkaline phosphatase.

No major structural alteration of alkaline phosphatase can be observed in the early stages of enzyme oxidative inactivation by the ascorbate model system. Fluorescence changes of protein-bound 8-anilino-1-naphthalenesulfonic acid suggest, however, that localized modifications take place. Oxidized alkaline phosphatase displays less catalytic efficiency (decrease of Vmax), while retaining the other kinetic properties, including the same affinity for substrates and inhibitors and the same activation energy of the native enzyme. Typical features of the modified protein are a decreased thermal stability and a biphasic heat inactivation profile, which make the oxidized form quite similar to aged enzymes. The lower response to Mg2+ activation indicates that the magnesium binding sites of alkaline phosphatase are probably the targets of the ascorbate system oxidative modifications.

Alkaline Phosphatase↗

Metergoline, naloxone, and sodium valproate did not modify arginine vasopressin response to insulin-induced hypoglycemia in man.

The present study was carried out in order to determine whether insulin-induced hypoglycemia exerts its stimulatory effect on plasma concentrations of arginine vasopressin (AVP) by interacting with a serotonergic, a GABA-ergic or an opioid pathway. For this purpose, the effect of the serotonergic antagonist metergoline (10 mg/day for 4 days po), the GABA-ergic agonist sodium valproate (600 mg in three divided doses po) and the opioid-receptor blocker naloxone (10 mg in a iv bolus) on the AVP response during an insulin (0.15 IU/kg bw) tolerance test (ITT) was evaluated in three groups of 6 normal men each. In all men, control ITTs were performed without drug treatments. Basal and ITT-stimulated AVP secretion was not modified by drug administration, suggesting that serotonergic, GABAergic and naloxone-sensitive opioid receptors are not involved in the regulation of AVP secretion in response to insulin-induced hypoglycemia.

Adrenocorticotropic Hormone↗

Early conformational changes and activity modulation induced by guanidinium chloride on intestinal alkaline phosphatase.

Moderate concentrations of guanidinium chloride induce both instantaneous and time-dependent modifications of the catalytic and optical properties of intestinal alkaline phosphatase, which undergoes consecutive conformational transitions at about 0.05 M, 0.25 M and 1.0 M denaturant. A paradoxical activation is observed up to 1.0 M-guanidine, with a maximum at 0.25 M- and a mid-point around 0.5 M-guanidine. Difference absorbance and fluorescence spectra imply a change in the state of ionization of the protein residues, with variation in molecular size suggested by light-scattering. Random-coil formation is indicated by a lower fluorescence yield, a more polar environment of the aromatic residues and another separate tryptophan emission. Iodide quenching confirms the alterations of conformation. Deprotonation favours the loss of the intramolecular constraints and the enhancement of the structure disruption by guanidine.

Alkaline Phosphatase↗

Alkaline phosphatase inactivation by mixed function oxidation systems.

Alkaline phosphatase is inactivated by mixed function oxidation systems. OH. radicals, generated via an ascorbate-modified Haber-Weiss cycle or a Fenton-type reaction, seem to be responsible for the protein oxidative damage. Experiments with hydroxyl radical scavengers, enzyme substrates, products, and metal cofactors suggest that a "site-specific" radical attack takes place at or near the active center. Vitamin E fails to protect alkaline phosphatase; uric acid, instead, is particularly effective in shielding the protein against covalent modifications.

Alkaline Phosphatase↗

The relationship between the optical properties and the kinetic behaviour of ascorbate-inhibited alkaline phosphatase.

Aromatic residues of bovine kidney alkaline phosphatase appear to be involved in the interaction with ascorbate, as shown by the strong quenching of intrinsic fluorescence and absorption. Difference u.v.-absorption spectra clearly indicate that conformational changes also occur. The pH value at which the greatest fluorescence deactivation is found is close to that necessary for optimal catalytic activity and for maximal inhibition by ascorbate. A protective effect against ascorbate is afforded by Pi. Time profiles of inactivation on one side and of absorbance and emission quenching on the other display opposite behaviours. Attempts to reverse the effects by the use of KOH fail to restore enzyme activity or to modify the spectral effects of ascorbate. The protein alterations are related, directly or indirectly, to the enzyme active centre and can be probably ascribed to the redox and chelating properties of ascorbate.

Alkaline Phosphatase↗

Ultrastructural observations in lichen nitidus.

Lichen nitidus (LN) and lichen planus (LP) are considered by some investigators to be two variants of the same disease, and by others to be two distinct dermatoses. In order to obtain further information about the relationship between LN and LP we examined the ultrastructure of lesions from two LN patients. In the central part of the lesion, the basement membrane was absent, or was interrupted by migrating phagocytes or lymphocytes. The basal cells and the lower cells of the stratum spinosum exhibited karyolysis and appeared to be compressed and often necrotic. In the upper dermis irregular cell debris full of clumps of tonofilaments and colloid-body-like structures was observed. A dense dermal infiltrate of macrophages, lymphocytes, fibrocytes, and Sezary-like cells was present. Signs of cooperation between lymphocytes and macrophages were also evident. The periphery of the lesion showed no pathological features except for enlargement of the intercellular spaces and the presence of mononuclear cells scattered through the epidermis. Several normal Langerhans cells were noticed. These ultrastructural findings were quite similar to those reported for LP.

Adult↗

Biochemical properties of alkaline phosphatase from endometrial cancer cells.

The occurrence of alkaline phosphatase (AP) activity was examined in several human endometrial adenocarcinomas. Catalytic activities were detectable only in 10 out of 15 tumors, with no apparent correlation between elevated AP and histological type. The apparent molecular weight of the enzyme after partial purification was about 140,000 daltons. Kinetic activity, thermodynamic properties and the pH dependence of the activity were in the ranges reported for other subforms. Several other physicochemical properties were also investigated and compared with those displayed by enzymes obtained from normal human tissues. The inhibition studies show that the enzyme shares several properties with the placental form, particularly in resistance to zinc chloride and EDTA action. On the other hand, in sensitivity to uncompetitive inhibitors and to urea and ascorbic acid, it is closer to other non-Regan heat-sensitive forms. The results support the view that a polymorphism in the expression of AP in neoplastic tissues can occur. A wider spectrum of physicochemical properties is clearly needed to define better the characteristics of oncodevelopmental enzymes.

Adenocarcinoma↗

Ascorbic acid stability in aqueous solutions.

Different water purity provokes a great variation of the stability of ascorbic acid and isoascorbic acid solutions. The effect of temperature on ascorbate aerobic oxidation was assessed by means of Arrhenius plots from which thermodynamic parameters were derived. The presence of bovine serum albumin drastically reduces the vitamin oxidation rate regardless of stereoisomerism. On the other hand the interaction with alkaline phosphatase, an enzyme inhibited by preincubation with vitamin C, does not modify significantly the stability in the experimental conditions used.

Alkaline Phosphatase↗

[Therapy with bromocriptine and behavior of various hormones in psoriasis patients].

The activity of bromocriptin has been clinically tested on 18 patients suffering from psoriasis. The plasma levels of Human Growth Hormone (HGH), Human Adrenocorticotropic Hormone (ACTH), Thyroid Stimulating Hormone (TSH), Prolactin (PL), Aldosterone and Cortisol were investigated in these 18 patients along with 35 untreated psoriatic patients and 19 normal subjects. Bromocriptin was shown to be effective in 13 of our 18 psoriatic patients. Plasma levels of HGH, ACTH, and Aldosterone, measured in all 53 psoriatic patients, were found to be higher than normal in 11, 26 and 40 patients respectively (HGH and Aldosterone: p less than 0,005; ACTH: p less than 0,001).

Adrenocorticotropic Hormone↗

[Cosmetic acne and a test of comedogenicity].

A comedogenic test was carried out on the internal ear canal of four adult, masculine, albino rabbits, using butter of cacao and linseed oil, both known to be present in various cosmetic products. Histologic observation after 14 days showed follicular hyperkeratosis conferming the validity of this test.

Acne Vulgaris↗

Characterization of alkaline phosphatase inactivation by ascorbic acid.

Ascorbic acid, isoascorbic acid and dehydroascorbic acid inhibit bovine kidney alkaline phosphatase activity. Ascorbic acid free radicals seem not to be involved. Dialysis does not make the inactivation reversible. A competitive mechanism can be inferred from experiments with phosphate and substrates, which block the activity decay. The influence of temperature, pH, other inhibitors and tertiary structure modifications on the inactivation process is also investigated.

Alkaline Phosphatase↗

[Differences in the culture of pigment cells from various sources].

In order to look for a new culture system suitable for investigating on melanocyte biology, in-vitro experiments have been carried out on pigmented basal cell carcinoma, pigmented seborrhoeic keratosis and melanocytic nevus. Melanocytes cultured from pigmented basal cell carcinoma and pigmented seborrheic keratosis are dendritic and DOPA positive. These cells can be considered similar to those of normal skin. Pigmented cells cultured from nevi are quite different. They are spindle-shaped and only to a little extent are DOPA positive. These findings suggest that nevus cells could be non mature melanocytes stopped at an intermediate stage of differentiation.

Basal Cell Carcinoma↗

[Use of monoclonal antibody 225.28S in the study of nevus cells in culture].

The aim of this work is to identify and purify cultured nevus cells. We used for our study monoclonal antibody 225.28S. This antibody reacts with a surface antigen which is expressed by nevus cells and melanoma cells, but it does not react with normal melanocytes. We studied 10 dermic nevi and we observed that antigenic determinant survives in cultured nevus cells. These results allowed us to employ the method of panning for purify cultured nevus cells.

Antibodies, Monoclonal↗

[Endothelial cell culture as a model for the study of wound healing].

Endothelial cells culture can be considered a reliable method for investigating about granulation tissue production in wound healing and for evaluating the pharmacological action of some chemicals on granulation tissue development. Endothelial cells have been obtained from human umbilical cords after trypsin treatment and their endothelial origin has been demonstrated by light microscopy, by immunofluorescence against factor VIII associated protein and by the platelet adhesion assay. The influence of fibronectin as substratum and of hyaluronic acid as soluble factor on adhesion and growth of endothelial cells has been investigated. Both these substances, but especially hyaluronate, determine a better attachment and an increase in the growth rate when compared with control cultures plated on plastic substratum and without hyaluronic acid in the culture medium.

Cell Adhesion↗

Spontaneous CSF rhinorrhea through the lamina cribrosa associated with primary empty sella.

Spontaneous CSF rhinorrhea associated with primary empty sella is a recently recognized condition. Because of its surgical indication, the preoperative evaluation of the possible site of the fistulous tract could be of fundamental importance. Most cases of primary empty sella reported in the literature have shown evidence of leakage through the sellar floor into the sphenoid sinus, thus requiring a transphenoidal approach. Non-traumatic defects in the anterior fossa are rarely reported in the literature. We have collected three new cases in which CT cisternography with non-ionic water-soluble contrast medium ruled out the possibility of a transellar fistula, thus suggesting a possible leak in the area of the lamina cribrosa. This was confirmed by the surgical outcome and by the follow-up of each patient.

Adult↗

Different effects of metoclopramide and domperidone on GH release in type I male diabetics.

Dopamine (DA) has a physiological role in the control of GH release from the pituitary. Studies have been carried out using DA agonist or antagonist, since in normal subjects DA does not cross the blood-brain-barrier (BBB). In contrast, the BBB is altered in type I diabetics, who respond to DA with a significant increase of GH release. In these patients Metoclopramide (MCP), an antidopaminergic drug, is also capable of stimulating GH release. In a previous paper, we suggested that this effect could be related to enhanced blood concentrations of DA, due to a reduced peripheral DA catabolism determined by MCP. However, since MCP crosses the BBB, an effect of this drug on other hypothalamic neurotransmitters could not be excluded. The purpose of the present study was to determine whether Domperidone (DOM) a drug which does not cross the BBB, but as well as MCP is thought to increase blood levels of DA, is also capable of inducing GH release in diabetics. Sixteen type I male diabetics were injected intravenously with MCP (6) or DOM (10) and a week later with normal saline. Ten normal subjects participated as controls to all three tests. MCP and DOM did not produce any effects in the normal controls; as expected, MCP induced a marked increase in serum levels of GH in the diabetics, while in contrast DOM did not stimulate GH secretion in diabetics. These data suggest that the effect of MCP is not due to the dopaminergic pathway previously described, but rather to a modulation of some other neurotransmitter at hypothalamic level, or to a direct effect on the pituitary in diabetics.

Adult↗