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Biomedical subjects

L Benedetti

Publications and source records attributed to L Benedetti.

At least 37 records · Page 2Linked to original sources

Detection of human herpesvirus 6 in plasma of children with primary infection and immunosuppressed patients by polymerase chain reaction.

A sensitive and specific polymerase chain reaction method for the detection of human herpesvirus 6 (HHV-6) DNA in serum or plasma has been developed. In total, 157 human serum or plasma samples were studied. HHV-6 DNA was detected in 6 (85.7%) of 7 children with exanthem subitum, 3 (23.1%) of 13 bone marrow transplant (BMT) recipients, 4 (22.2%) of 18 human immunodeficiency virus (HIV)-infected patients, 1 (2.6%) of 39 patients with chronic fatigue syndrome, and none of 37 healthy adults. In the HHV-6-positive BMT recipients, HHV-6 plasma DNA was transiently detected during episodes of fever and respiratory infection. In children with exanthem subitum and in 1 HIV-infected patient, the HHV-6 strains were characterized as variant B, whereas variant A was detected in all other patients. Detection of viral DNA in serum or plasma is a marker of active infection that can be used to investigate the role of HHV-6 in human disease.

Adult↗

Arginine-vasopressin induces differentiation of skeletal myogenic cells and up-regulation of myogenin and Myf-5.

The neurohypophyseal nonapeptide arginine8-vasopressin (AVP) induces phosphoinositide turnover and calcium and pH changes in skeletal myogenic cells in culture. In order to investigate the effect of AVP on skeletal myogenesis, we examined the effect of this hormone on proliferating mononucleated L6 myoblast cultures. Addition of AVP to the medium resulted in the formation of much larger myotubes than those formed in its absence and in a significant increase (2.2-fold) of the percentage of fusion within 3-4 days of treatment. The effect of AVP was dose dependent, in the 10 nM to 1 microM range, and was observed also in primary cultures of mouse satellite cells. The rate of growth of L6 cells was not affected by AVP treatment. The induction of morphological differentiation by AVP correlated with an increased expression of muscle-specific gene products, such as myosin, and an increased number of acetylcholine receptor sites. The accumulation of mRNA transcripts of the acetylcholine receptor subunits was also enhanced by AVP. The mechanism involved in the myogenic action of AVP was investigated. Using AVP-related peptides and antagonists, we found that a specific chemical structure is required and that V1 receptors probably mediate the effect on myogenesis. Expression of muscle-specific transcription factor genes Myf-5 and myogenin and their products are strongly upregulated by AVP. Our findings support the hypothesis that AVP may represent a novel physiological modulator of skeletal muscle differentiation through its action on muscle regulatory genes.

Amino Acid Sequence↗

Characterization of a phenobarbital-inducible Ah receptor-like protein in the Sprague-Dawley rat.

Immature male Sprague-Dawley rats were treated with phenobarbital (PB) at a dose of 50 mg/kg for 3 successive days and then sacrificed 1, 6, or 9 days after the last treatment. An increase in the concentration of hepatic cytosolic proteins able to bind specifically to [3h]-2,3,7,8-tetrachlorodibenzo-p-dioxin ([3H])TCDD) was observed. The level of the induced protein fell in parallel with the elimination of PB in vivo. Readministration of PB 9 days after the last treatment, by which time only the constitutive receptor was present, induced additional protein at levels similar to those obtained upon the original administration of the drug. There was considerable variability within a given treatment group in both the ratio of constitutive to induced protein and the total amounts of these proteins. The kinetic properties of the induced protein were significantly different from those of the constitutive Ah receptor. In particular, [3H]TCDD was released faster from the liganded induced protein than from the constitutive Ah receptor. The protein induced by PB had properties very similar to those of the protein induced by a combination of 2,3,7,8-tetrachlorodibenzo-p-dioxin and 2,2',4,4',5,5'-hexachlorobiphenyl.

Animals↗

Biodegradation of hyaluronic acid derivatives by hyaluronidase.

Hyaluronic acid (salt) (HA) has been chemically modified as a biomaterial for medical applications such as controlled drug release matrices, nerve guides and wound dressings. A series of HA derivatives, which include different ester types and different degrees of esterification, have been used to investigate the stability of these materials in testicular hyaluronidase. Gel permeation chromatography and capillary viscometer have been employed to determine the size of the molecules, the former used for the water insoluble derivatives that dissolve in dimethyl sulphoxide, the latter for the water soluble samples. The preliminary experimental results indicated that the molecular weight of fully esterified hyaluronic acid (both ethyl and benzyl esters) did not decrease after treatment in the enzyme for 7 and 14 days while the water soluble partially esterified HA were degraded by the enzyme producing a sharp reduction of viscosity within minutes. These observations tend to suggest that the carboxylic groups in the beta-glucoronic acid unit are the activation centre of this enzyme and the total blockage of these groups can restrict the cleavage of beta (1-->4) glycoside bonds by this enzyme.

Biocompatible Materials↗

Changing features of proximal vein thrombosis over time.

Recently, the sensitivity of impedance plethysmography (IPG) for the diagnosis of acute deep-vein thrombosis (DVT) in symptomatic outpatients has been questioned. In order to verify whether a change in the venographic pattern of DVT has occurred over years, accounting for the decreased sensitivity of IPG, the authors compared two series of consecutive venograms demonstrating proximal DVT, performed between 1984-1988 (166 patients) and 1990-1992 (140 patients). They evaluated both the extension and the occlusiveness of deep-vein thrombi in the two series. Moreover, changes in the referral characteristics of patients were investigated. In the second series of venograms a significant decrease in thrombi extension, expressed by a lower prevalence of iliac vein thrombosis (29% versus 43%; P = 0.0074) was observed; moreover, a significant increase in the prevalence of nonocclusive thrombi (22% versus 8%; P = 0.0004) was also recorded in the second series when compared with the first. During the study period, among the referral characteristics of patients, the authors observed both a significant decrease in the prevalence of proximal DVT (from 31% to 24%; P < 0.01) and a slight and not statistically significant decrease in the median time elapsed between onset of symptoms and referral for objective testing (from eight and a half to seven days). In conclusion, proximal deep-vein thrombi are currently less extensive and occlusive than observed in the past. These results might depend on earlier referral of less symptomatic patients and might explain the recently reported decrease in IPG sensitivity for proximal DVT.

Analysis of Variance↗

Human neutrophil chemokinesis and polarization induced by hyaluronic acid derivatives.

Neutrophils and macrophages are known to undergo significant modifications in their morphology and basal metabolism in response to chemical factors, in particular changes in the shape, movement, phagocytic activity and degranulation. These phenomena often involve an increase in chemokinesis and cellular secretory activity, usually expressed in antimicrobial activity. Once activated, the cells can move quickly towards the source of the stimulus, where they produce and release great amounts of enzymes (e.g. proteases, hydrolases, lysozyme) and reactive oxygen metabolites (e.g. O2-., H2O2, OH.). This study has examined the ability of surfaces of selected biomaterials to influence neutrophil morphology and locomotion. The surface of two films derived from hyaluronic acid derivatives were compared with that of glass. The two hyaluronic acid derivatives, despite having a similar chemical structure, were shown to interact with human neutrophils in different ways. A hyaluronic acid ethyl ester stimulated the whole population of neutrophils to take up a non-spherical morphology (polarize) and to move with a velocity similar to that of N-formyl-methionine-leucine-phenylalanine-stimulated cells on a glass surface. In contrast, only 44% of the examined cells on the surface of hyaluronic acid benzyl ester were polarized and their mean speed was only slightly higher with respect to that found with non-stimulated cells on glass. Moreover, while on the benzyl ester and on glass a correlation between neutrophil circularity (i.e. the shape of the cell) and cell speed was found, the ethyl ester did not show any correlation.

Biocompatible Materials↗

Biocompatibility and biodegradation of different hyaluronan derivatives (Hyaff) implanted in rats.

Hyaluronan (HL), a naturally occurring glycosaminoglycan, has been chemically modified through the esterification of its carboxylic groups with different types of alcohol. The physico-chemical properties of these new biopolymers allow the preparation of many biomaterials which may be used in several medical applications. In the present study both the biocompatibility and biodegradation of some water-insoluble HL esters have been evaluated, either as raw material or as manufactured devices after subcutaneous and intraperitoneal implantation in male rats. The inflammatory response and the degree of resorption for each tested material are reported. The relationships between the degree of esterification and the type of alcohol used with the above parameters are also investigated.

Animals↗

A simple ultrasound approach for detection of recurrent proximal-vein thrombosis.

BACKGROUND: The objective of this study was to develop a simple ultrasound method for measuring thrombus regression in patients with proximal deep-vein thrombosis (DVT) and to test its utility for the detection of DVT recurrence. METHODS AND RESULTS: The study comprised a cross-sectional survey and a prospective investigation (149 and 145 patients, respectively). In both phases, the normalization rate of a previously abnormal ultrasound test, applying the criterion of full compressibility of the common femoral and popliteal veins (C-US method), was assessed. In the prospective study, the vein diameter under maximum compression (thrombus thickness) was measured in the abnormal venous segments at scheduled times (1, 3, 6, and 12 months). In patients presenting with suspected DVT recurrence, the procedure was repeated and results were compared with those available from the previous examination. Noncompressibility of a previously normal(ized) venous segment and enlargement of thrombus thickness (> or = 2 mm) were considered diagnostic of proximal DVT recurrence. The diagnostic accuracy of the C-US method alone, as well as of the combined ultrasound methods (C-US + thrombus thickness), was assessed against contrast phlebography. C-US test normalization occurred in only 30% of patients within 1 year. A significant reduction of the thrombus mass (P < .0001) was recorded throughout the entire study period. However, a major decrease in thrombus mass (> 50%) was recorded within the first 3 months. Of 29 patients who developed a suspected recurrent DVT, phlebography confirmed diagnosis in 11. The C-US method alone showed an excellent accuracy (100%) but was applicable in only 6 patients (21%). Both the sensitivity and the specificity for proximal DVT recurrence of the combined ultrasound methods were 100% (95% confidence interval, 69% to 100% and 81% to 100%, respectively) and were applicable in all patients. CONCLUSIONS: The serial ultrasound measurement of thrombus mass after an acute episode of DVT may allow the correct identification of patients who develop a recurrent proximal-vein thrombosis.

Cross-Sectional Studies↗

[Achilles tendon ossification and achillodynia].

Achilles tendon ossification is a rare and asymptomatic condition. The possible sequelae of tendinitis, bursitis and fracture are some of the causes of achillodynia. Achillodynia is often unrelieved by conservative management and excision of the ossification may be necessary. Moreover, ossification were found in 3 of 41 patients with spontaneous rupture of the Achilles tendon. Predisposing degenerative changes of the tendon were found in all patients. Thus there is no evidence that ossification by itself may increase the risk of tendon rupture.

Achilles Tendon↗

Synthesis and preliminary characterisation of new esters of the bacterial polysaccharide gellan.

Under the appropriate experimental conditions, ethyl, propyl, and methylprednisolon-21-yl esters of gellan can be obtained without significant degradation. At low degrees of esterification (de), depending on the ester moiety, the products are water-soluble, which allows the influence of hydrophilicity and charge density on their ability to assume an ordered conformation in dilute aqueous solution to be studied. With high de, the products were soluble only in organic solvents (e.g., methyl sulphoxide) with good film-forming capacity. The methylprednisolon-21-yl esters have been characterised in a preliminary manner in terms of drug-release kinetics.

Alcohols↗

Prosomatostatin is processed in the Golgi apparatus of rat neural cells.

Proteolytic processing of somatostatin precursor produces several peptides including somatostatin-14 (S-14), somatostatin-28 (S-28), and somatostatin-28 (1-12) (S-28(1-12)). The subcellular sites at which these cleavages occur were identified by quantitative evaluation of these products in enriched fractions of the biosynthetic secretory apparatus of rat cortical or hypothalamic cells. Each of the major cellular compartments was obtained by discontinuous gradient centrifugation and was characterized both by specific enzyme markers and electron microscopy. The prosomatostatin-derived fragments were measured by radioimmunoassay after chromatographic separation. Two specific antibodies were used, allowing the identification of either S-28(1-12) or S-14 which results from peptide bond hydrolysis at a monobasic (arginine) and a dibasic (Arg-Lys) cleavage site, respectively. These antibodies also revealed prosomatostatin-derived forms containing at their COOH terminus the corresponding dodeca- and tetradecapeptide sequences. Whereas the reticulum-enriched fractions contained the highest levels of prosomatostatin, the proportion of precursor was significantly lower in the Golgi apparatus. In the latter fraction, other processed forms were also present, i.e. S-14 and S-28(1-12) together with the NH2-terminal domain (1-76) of prosomatostatin (pro-S(1-76). Inhibition of the intracellular transport either by monensin or by preincubation at reduced temperature resulted in an increase of prosomatostatin-derived peptides in the Golgi-enriched fractions. Finally, immunogold labeling using antibodies raised against S-28(1-12) and S-14 epitopes revealed the presence of these forms almost exclusively in the Golgi-enriched fraction mainly at the surface of saccules and vesicles. Together these data demonstrate that in rat neural cells, prosomatostatin proteolytic processing at both monobasic and dibasic sites is initiated at the level of the Golgi apparatus.

Animals↗

Prosomatostatin II processing is initiated in the trans-Golgi network of anglerfish pancreatic cells.

Anglerfish prosomatostatin II, the precursor of somatostatin-28 II, is produced in different cells from prosomatostatin I, by a cleavage at Arg73. Antibodies were raised against the carboxy-terminal [64-72] portion of the precursor II upstream from somatostatin-28 II sequence. These antibodies recognized only this epitope when unmasked from the entire precursor, allowing the detection of the [1-72] domain which was isolated from pancreatic islets extracts. The antibodies were used to monitor the peptide bond cleavage occurring at the carboxy terminus of Arg73 to generate somatostatin-28 II. Immunocytochemistry revealed labeling both in the vesicles budding from the trans-Golgi network and in the dense core granules. Together, these data support the conclusions that i) prohormone processing is initiated in the Golgi apparatus of the pancreatic islet cells; ii) the "non-hormonal" [1-72] amino-terminal domain of the precursor may be involved in some intra and/or extra-cellular function(s).

Animals↗