Biomedical subjects

B Pozzetto

Publications and source records attributed to B Pozzetto.

At least 73 records · Page 4Linked to original sources

PubMed1994

Arbitrarily primed PCR, ribotyping, and plasmid pattern analysis applied to investigation of a nosocomial outbreak due to Enterobacter cloacae in a neonatal intensive care unit.

In December 1992, Enterobacter cloacae was isolated from the oropharynx and respiratory tract of six ventilated neonates hospitalized in the intensive care unit (ICU) of our hospital. To establish the spread of the outbreak, 41 strains of E. cloacae were analyzed for genotypic markers by three methods: plasmid profile analysis, ribotyping with EcoRI or PvuII endonuclease, and arbitrarily primed (AP) PCR. The tested strains included 12 isolates from the 6 epidemic cases, 4 isolates from the respiratory tract of 4 children hospitalized in other wards during the same period, 13 isolates from 12 children hospitalized in pediatric units before or after the outbreak, and 12 epidemiologically unrelated isolates. Ribotyping and AP PCR demonstrated that each of the last 12 strains exhibited distinct genomic patterns, as did each of the strains isolated from neonates hospitalized before or after the epidemic peak. Conversely, two clones of strains were found among the isolates recovered in December, with concordant results being obtained by the three typing methods: the first clone included seven strains from five ventilated children in the ICU and two children from another ward; another clone was shared by one neonate in the ICU and an infant from another ward. These results indicate that ribotyping and AP PCR-the latter applied, to our knowledge, for the first time to the genotypic analysis of E. cloacae--represent very discriminatory tools for the investigation of nosocomial outbreaks caused by this species.

Bacterial Typing Techniques↗

PubMed1994

Is there IgA of gut mucosal origin in the serum of HIV1 infected patients?

This study was performed in 77 HIV1 seropositive adult patients to characterise the IgA hyperglobulinaemia seen in the serum during the course of HIV infection. It was shown that both IgA1 and IgA2 subclass concentrations were simultaneously increased but the IgA1 increase was predominant. Secretory IgA (SIgA) concentration was significantly increased and IgA activity to gliadin, bovine serum albumin, and casein could be detected and was correlated with SIgA concentration. In contrast, IgA activity to cytomegalovirus and to tetanus toxoid did not correlate with total IgA concentration. These data suggest the presence of IgA from gut mucosal origin in the serum of these patients. Hyper IgA was inversely correlated with the CD4+ cell number. The increase of all parameters studied varied according to the total IgA concentration in the serum but was also directly related to the stage of immune deficiency in patients with hyper IgA.

Acquired Immunodeficiency Syndrome↗

PubMed1994

[Immunoglobulins E and human immunodeficiency virus infection (HIV)].

Except for Ig E, serum immunoglobulin abnormalities in persons with human immunodeficiency virus (HIV) infection have been well described. Serum IgE levels have been shown to rise with progressive disease. The authors evaluated IgE in 148 HIV-seropositive individuals with or without acquired immunodeficiency syndrome (AIDS). Mean serum IgE levels were compared between groups based on absolute CD4 lymphocyte counts or clinical status (CDC) and with a seronegative control group. Higher serum IgE levels were observed in seropositive-patients. A rise in IgE serum is common in patients with HIV infection; it could be link with an earlier dysregulation in the IgE synthesis. No correlation was found between IgE level and CD4 counts.

Adolescent↗

PubMed1993

Is there a relationship between malnutrition, inflammation, and post-vaccinal antibody response to influenza viruses in the elderly?

This study was designed to explore the relationship between malnutrition, inflammation, and the specific antibody response after influenza vaccination in the elderly. Eighty-two aged subjects, immunized annually against influenza with a trivalent inactivated vaccine, were evaluated for 9 protein markers (albumin, thyroxin-binding prealbumin, transferrin, immunoglobulins (Ig) G, M, and A, orosomucoid, haptoglobin, and C reactive protein) and for their antibody response to influenza viruses in comparison to 29 younger adults who received the same vaccine and 21 unvaccinated adults. IgM and nutritional markers were significantly reduced in the aged as compared to controls, while the opposite pattern was seen for IgA and inflammatory markers. No difference was observed between the elderly and the controls with regard to the antibody response to influenza virus after vaccination. Reciprocally, influenza immunization had no influence on the levels of the protein variables. These results suggest that the protein status does not play an important role in the antibody response to influenza vaccination in the elderly, a fact which could be related to the slight involvement of cellular immunity in the defense against influenza reinfection.

Adult↗

PubMed1993

Genotypic homogeneity of nosocomial Pseudomonas aeruginosa O12 strains demonstrated by analysis of protein profiles, DNA fingerprints and rRNA gene restriction patterns.

The spread in Europe of a single multiresistant strain of Pseudomonas aeruginosa serotype O12 has been suggested. This bacterium was responsible for a nosocomial outbreak in our hospital in 1988-1989. Three different epidemiological methods were used to analyze 30 strains isolated during five consecutive years. Protein profile analysis and chromosomal DNA fingerprinting with four different enzymes revealed closely related patterns. rRNA gene restriction fragment length analysis performed with a digoxigenin-labelled probe showed identical hybridization patterns with four to six bands according to the endonuclease used. Combination of the three typing methods showed genotypic homogeneity of these Pseudomonas aeruginosa O12 strains, despite a relative increase in their antibiotic resistance.

Bacterial Proteins↗

PubMed1993

Ventilator temperature sensors: an unusual source of Pseudomonas cepacia in nosocomial infection.

A prospective study was undertaken to determine the source of Pseudomonas cepacia colonization and infection that had affected ventilated patients in an Intensive Care Unit (ICU) for three years. Thirty-eight patients undergoing mechanical ventilation were enrolled during a six-week period. Samples were taken from patients, ventilator circuits and the environment for culture. P. cepacia was isolated from the condensate formed in the ventilator circuit and the source of the contamination was shown to be the temperature sensor. Ribotyping of the representative strains of P. cepacia performed with two endonucleases, EcoRI and PvuII, confirmed the homogeneity of the isolates from patients and ventilator circuits. A modification of the procedure for disinfection of the temperature sensors resulted in the eradication of P. cepacia from the ICU.

Burkholderia cepacia↗

PubMed1993

[Enterovirus infections and systemic clinical manifestations with prolonged inflammatory syndrome: association with a persistence of specific IGM antibodies].

We report 9 cases of enteroviral infection associated with systemic inflammatory disease (including 4 cases of vasculitis, 1 case of periarteritis nodosa, 1 case of Sharp's syndrome). We then reviewed 36 cases of enteroviral infection with persistent IgM antibodies diagnosed in the virology laboratory in a 6-year period: among them 11 cases were found to present with subacute or chronic inflammatory disease. We conclude that enteroviruses might be important triggers of systemic inflammatory disease.

Enterovirus Infections↗

PubMed1993

Prognostic value of serum immunoglobulin A antibodies to pol gene products during HIV-1 infection.

Serum specimens from 66 HIV-1-infected subjects were tested by ELISA for the presence of IgA antibodies to HIV-1: 44 samples were found positive and 37 were confirmed by immunoblot. In these subjects, the presence of anti-HIV IgA antibodies was studied in relation to the total count of circulating CD4+ lymphocytes and to the level of serum IgA. A significative correlation (P < 0.03) was found between the absence of IgA to the subunit p68 of the reverse transcriptase and a count of CD4+ cell < 400/mm3 or total IgA level over 4.25 g/l. The same pattern was observed for the IgA antibodies to the p52 subunit but the association was just not significant (P < 0.07). No significant decrease was noted for the IgA directed towards the other proteins of HIV-1, especially the products of the gag gene.

Adult↗

PubMed1993

Transient erythroblastopenia of childhood presenting with echovirus 11 infection.

We report a case of transient erythroblastopenia in a three-year-old girl presenting with echovirus 11 infection. Viral infection was demonstrated by isolation of echovirus 11 in stool cultures and the presence of echovirus 11-specific IgM antibody in serum. We suggest that echovirus may have played a role in the pathogenesis of transient erythroblastopenia of childhood in this patient.

Anemia↗

PubMed1993

Characterization of unrelated strains of Staphylococcus schleiferi by using ribosomal DNA fingerprinting, DNA restriction patterns, and plasmid profiles.

The molecular characteristics of 31 unrelated strains of Staphylococcus schleiferi isolated from 13 hospitals between 1973 and 1991 were determined by ribosomal DNA fingerprinting by using a digoxigenin-labeled DNA probe, genomic DNA restriction patterns, and plasmid profiles. Only six strains harbored one or two plasmids. DNA restriction analysis, which was carried out with five endonucleases (EcoRI, HindIII, PstI, PvuII, and ClaI), did not allow us to discriminate between isolates. Ribotyping with HindIII, ClaI, or EcoRI enzymes generated six, seven, and nine distinct patterns, respectively. With the combination ClaI-EcoRI, 13 ribotypes were obtained among the 31 strains, suggesting a relative heterogeneity within the species. Moreover, all strains shared two or three common bands, according to the endonuclease used, which were relatively specific for S. schleiferi in comparison with the ribosomal banding patterns described for other coagulase-negative staphylococci. These results illustrate that ribotyping can be used for the epidemiological investigation of S. schleiferi isolates and possibly for taxonomic analysis in this species.

DNA Fingerprinting↗

PubMed1993

Role of influenza vaccination in the elderly during an epidemic of A/H1N1 virus in 1988-1989: clinical and serological data.

During a winter epidemic of A/H1N1 influenza virus, we evaluated the protection conferred by vaccination of 285 residents of a nursing home. Fifteen of 204 members of the nursing staff were also vaccinated. Serological determinations were performed before and after vaccination using radial hemolysis (RH) and neuraminidase inhibition (NI) tests. In the outbreak period, only one influenza case was noted in the vaccinated elderly and none among the vaccinated nursing staff. On the other hand, 38 cases (20%) occurred in the unvaccinated hospital personnel. Twenty-one percent of the elderly people exhibited seroconversion to the vaccinal strain by RH and NI while 27 and 20% of the vaccinated nursing staff seroconverted by the same tests, respectively. Thus, the clinical protection conferred by influenza vaccination was excellent and much greater than expected from serological results.

Aged↗

PubMed1992

Antibodies to Epstein-Barr virus and cytomegalovirus in relation to CD4 cell number in human immunodeficiency virus 1 infection.

Interaction between herpesviruses and human immunodeficiency virus (HIV)1 is postulated in the progression of HIV disease. In order to evaluate the specific antibody responses directed to Epstein-Barr virus (EBV) and cytomegalovirus (CMV) and to provide serological evidence suggesting reactivation of these viruses able to accelerate the immunodeficiency, we studied IgA and IgG titres to EBV and CMV in the serum of HIV positive patients in relation to the CD4 cell number. The titres of IgG antibodies to EBV and the prevalence of IgG to CMV were significantly higher in HIV positive patients compared to control high risk HIV negative subjects. In HIV infected patients, anti-VCA IgG antibodies increased and anti-EBNA IgG antibodies decreased progressively in relation to the decline of CD4 cell number whereas anti-CMV IgG antibodies did not varied significantly at the same time. Anti-VCA IgA and anti-EA IgG antibodies were found uncommonly and with low titres. IgA antibodies to EA and CMV were not detected in any patient. The variations in EBV antibody response that we describe in HIV infection were previously reported in other immunodeficiency states and could be distinctive of these diseases.

Acquired Immunodeficiency Syndrome↗

PubMed1992

Evidence for T-cell involvement during the acute phase of echovirus meningitis.

Little is known about the cellular immune response during the acute phase of enterovirus infection. This was studied in patients with echovirus meningitis by analysing changes in the lymphocyte subset distribution both in blood and in cerebrospinal fluid (CSF) and their stage of activation. A significant increase of whole T-cell and CD4+ T-cell counts was observed in CSF in parallel with a slight decrease of T-cell percentage in blood. Activation of the cellular immune response is supported by the observation of elevated neopterin levels in serum and CSF. However, the phenotypic markers of T-cell activation, IL-2 receptor (CD25), and HLA-DR antigens were not detected in blood or in CSF.

Adolescent↗

PubMed1992

Evaluation of four methods for rapid detection of adenovirus.

Four methods for rapid detection of adenovirus were evaluated by testing retrospectively 28 frozen clinical specimens from which an adenovirus strain had been isolated. After thawing all specimens were retested for the presence of adenovirus by conventional culture on KB cells and found to be positive. The four tests used for rapid detection of adenovirus were a 48-hour culture technique, and an immunoassay, a latex agglutination test and an immunofluorescence assay for direct detection of viral antigen using commercially available reagents. Of the 28 specimens all were positive in the 48-hour culture, 25 (89%) positive in the immunoassay and 10 (36%) positive in the latex agglutination test. Six of eight nasopharyngeal aspirate specimens were positive in the immunofluorescence assay. Twenty-five clinical specimens negative for adenovirus on conventional culture were also negative in the 48-hour culture technique. Overall, the rapid (48-hour) culture technique was 100% sensitive and 100% specific compared to conventional culture. The direct detection of viral antigen by immunoassay was less sensitive, however results were available within a few hours. Prospective comparative studies are warranted to determine whether these rapid techniques could replace conventional culture in the routine diagnosis of adenovirus infection.

Adenoviridae↗

PubMed1992

Comparison of excretory/secretory and circulating antigens of Toxoplasma gondii by enzyme immunoassay and immunoblotting.

Toxoplasma gondii trophozoites (RH strain) were cultured in embryonic fibroblasts in order to study the kinetics of production of excretory/secretory antigens, and the results were compared to the production of circulating antigens in an in vivo mouse model. By capture-ELISA, excretory/secretory antigens were first detected on the fourth day of culture whereas circulating antigens were first detected 1 day after infection. Similar concentrations of antigens were detected in both models as evidenced by comparable absorbance values. By immunoblotting, the excretory/secretory antigens were also detected later compared to circulating antigens (day 4 vs day 1). Seven major polypeptides were detected in both antigen preparations, six of them having the same molecular mass (110, 75, 48, 30, 24 and 22 kDa).

Animals↗

PubMed1992

A 44,000 glycoprotein is involved in the attachment of echovirus-11 onto susceptible cells.

Cellular receptors play an important role in viral pathogenesis. Until now little was known on echovirus (EV) receptor. Using detergent-treated KB cell extracts as immunogen, a mouse monoclonal antibody (Mab 143) was produced that selectively blocks the attachment of EV-11 to KB and other susceptible cells. By immunoblotting, Mab 143 detected a 44,000 protein on susceptible cell lines but not on cell lines from nonprimate origin. The receptor protein complex, purified from KB cell membranes by immunoaffinity using Mab 143 as ligand, was shown to contain a single glycoprotein with apparent molecular weight of 44,000 (gp44). The role of gp44 in the attachment of EV-11 onto KB cells was demonstrated by the ability (i) of affinity-purified gp44 to reduce the infectivity of EV-11 and (ii) of rabbit polyclonal antisera raised against gp44 to protect cells from the replication of various EV, as did Mab 143.

Animals↗

PubMed1992

Emergence of quinolone-imipenem cross-resistance in Pseudomonas aeruginosa after fluoroquinolone therapy.

Emergence of resistance to fluoroquinolones was observed in two clinical isolates of Pseudomonas aeruginosa after ciprofloxacin or norfloxacin monotherapy. In the first case, the resistant variants exhibited quinolone-imipenem cross-resistance (MIC of norfloxacin and ciprofloxacin: 16 mg/L; MIC of imipenem: 8 mg/L), although the patient had never received imipenem treatment, while the strain from the second case remained imipenem-susceptible (MIC of norfloxacin or ciprofloxacin: 8 mg/L; MIC of imipenem: 2 mg/I). The frequency of in-vitro emergence of variants resistant to imipenem and fluoroquinolones was studied for the two strains, with imipenem or fluoroquinolones as selecting agents. Ciprofloxacin and three other quinolones (norfloxacin, temafloxacin and tosufloxacin) selected imipenem-resistant variants in a similar way to imipenem for the first strain, but not for the other. In contrast, imipenem did not select quinolone-resistant variants from either strain. For both strains, killing curves demonstrated that a bactericidal effect could be obtained with a drug combination (2 x MIC of ciprofloxacin and 2 x MIC of imipenem) without any selection of resistant mutants after 24 h, thereby suggesting the possible use of this combined regimen for treating severe P. aeruginosa infection.

Anti-Infective Agents↗

PubMed1992

Immunoblotting patterns with Mycoplasma pneumoniae of serum specimens from infected and non-infected subjects.

Two hundred and ninety-four serum specimens from 248 subjects, whose complement fixation (CF) titres to Mycoplasma pneumoniae were known, were further investigated by IgG immunoblotting. After analysis of M. pneumoniae proteins by SDS-PAGE, nine polypeptides (p) with mol. wts of 180-43 Kda were selected for immunoblotting studies. Antibodies to M. pneumoniae measured by immunoblotting appeared progressively with age; most subjects more than 19 years old gave positive results. For most of the polypeptides, there was an increase in the frequency of band detection when the CF titres were higher. Furthermore, paired serum specimens from 10 patients with M. pneumoniae infection, as demonstrated by a rise in CF antibody titre, were tested for IgG blotting patterns. Generally, p180 (the P1 adhesin of M. pneumoniae), p172 and p84 were shown to be the dominant targets of the immune response to this organism and may have diagnostic value.

Adolescent↗