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Biomedical subjects

G D Rottini

Publications and source records attributed to G D Rottini.

At least 19 recordsLinked to original sources

Physical characterization of the pore forming cytolysine from Gardnerella vaginalis.

The cytolytic toxin (CTox) produced by Gardnerella vaginalis is able to form voltage-dependent cationic channels when incorporated in lipid membranes (Moran et al. (1991) FEBS Lett. 283, 317-320). Osmotic protection experiments show that toxin incorporated in human erythrocytes forms pores between 18 A and 28 A in diameter. A hypothesis of pore formation as a primary event to produce cytolysis is proposed. The CTox activity increases when cells are depolarized by increasing the extracellular K+ concentration, probably reflecting the voltage dependent character of CTox formed channels. The cytolytic effect of the toxin was prevented by low temperatures and was a function of the extracellular Ca2+ concentration, suggesting a Ca2+ influx as part of the lytic mechanism. Binding of CTox to erythrocytes was dependent on external Ca2+ and was less temperature-dependent. Dose-response analysis suggests cooperativity of the toxin for the lytic activity, although no direct evidence of oligomerization has been found.

Blotting, Western↗

A new method for assessment of serum-induced damage to E. coli.

A simple and rapid spectrophotometric assay for the kinetic evaluation of serum-induced damage to E. coli is described, based on changes in the optical density (OD) of a bacterial suspension. Exposure of antibody-coated E. coli to human absorbed serum results in a diphasic response, namely an increase in OD, which reaches a maximum at about 17 min and is followed by a progressive decrease in OD until a minimum value is reached after 45 min. The increase and the decrease in OD are related to bacterial death and bacterial lysis, respectively.

Blood Bactericidal Activity↗

Incidence of myeloperoxidase deficiency in an area of northern Italy: histochemical, biochemical and functional studies.

Forty-five subjects with a complete deficiency of myeloperoxidase were identified in an area of the region Friuli-Venezia Giulia in north-eastern Italy using the Hemalog D system as the screening technique. Histochemical and biochemical tests performed on the leucocytes of some of these subjects confirmed the defects shown by the Hemalog D system. The defect was of genetic origin in seven subjects. The genetic origin could be suspected in another eight subjects since more than two affected members were present in a given family. Eosinophil peroxidase, which is present in MPO deficient subjects, interfered with the guaiacol assay of MPO, and in several cases masked the genetic transmission. An assay was developed using o-dianisidine as the electron donor which considerably reduced the interference by EPO. With this assay an autosomal recessive pattern of inheritance was found. The MPO deficient leucocytes had a higher respiratory burst than control cells and an impaired bactericidal activity, at early post-phagocytic periods, which became comparable to that of control cells at later stages. Particle ingestion by the MPO-deficient cells was normal.

Blood Bactericidal Activity↗

Effect of antibodies and complement on the interaction between Escherichia coli 0111:B4 and polymorphonuclear leukocytes.

The interaction of Escherichia coli 0111:B4 with polymorphonuclear leukocytes in the presence of specific antibodies and complement was studied. This strain, which is resistant to phagocytosis by polymorphonuclear leukocytes, may be ingested and killed by the phagocytes in the presence of both antibodies and fresh serum. The ineffectiveness of fresh serum to promote ingestion of E. coli 0111:B4 by the phagocytes in the absence of antibodies reflects the inability of this strain to activate the complement system through the alternative pathway. Investigation of the mechanisms of the bacterial killing by polymorphonuclear leukocytes showed that both antibodies and complement were required for the oxygen-independent bactericidal system, whereas they were not needed for the oxygen-dependent system.

Antibodies↗

[Pathogenicity of E. coli: biological aspects and problems related to laboratory diagnosis (author's transl)].

The biological properties which may contribute to the pathogenicity of E. coli are reviewed in this article. Specifically the following topics are discussed in detail: 1) adhesion to the intestinal epithelial cells, 2) production of enterotoxins, 3) invasiveness of and ability to multiply within the epithelial cells, 4) insensitivity to complement lysis or inability to activate the alternative pathway of complement, 5) resistance to phagocytic killing. The various techniques which might be of potential usefulness in the clinical laboratory to test the parameters of the pathogenicity of E. coli are briefly outlined. The dissociation between the definition of pathogenicity, as established on the basis of the results of E. coli serotyping, and the criteria of pathogenicity listed above is brought to the attention of the reader. As specificity regards the toxinogenicity, what emerged from a survey of the literature, was that only one of the so called "enteropathogenic" strains of E. coli, according to the serotype classification, was found to produce an enterotoxin.

Clinical Laboratory Techniques↗