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Biomedical subjects

M Donati

Publications and source records attributed to M Donati.

At least 37 records · Page 2Linked to original sources

Comparative in-vitro activity of moxifloxacin, minocycline and azithromycin against Chlamydia spp.

The in-vitro activity of moxifloxacin, a new 8-methoxyquinolone, was compared with minocycline and azithromycin against 40 strains of Chlamydia trachomatis, Chlamydia pneumoniae and Chlamydia psittaci. Both the MIC and the MBC of moxifloxacin ranged from 0.03 to 0.125 mg/L. MICs of minocycline ranged from 0.015 to 0.06 mg/L and MBCs between 0.03 and 0.25 mg/L. MICs of azithromycin ranged from 0.03 to 0.125 mg/L and the MBCs between 0.06 and 0.5 mg/L. MBC values of moxifloxacin were the same as MICs in 32 (80%) of 40 strains tested, whereas those of minocycline and azithromycin were two to four times higher than their MICs. These data confirm those previously obtained indicating that quinolones kill chlamydial strains at concentrations equivalent to their MICs.

Anti-Bacterial Agents↗

PCR-based restriction pattern typing of the vacA gene provides evidence for a homogeneous group among Helicobacter pylori strains associated with peptic ulcer disease.

The results of PCR-based molecular typing of Helicobacter pylori strains by restriction fragment length polymorphism analysis of a 1, 161-bp nucleotide sequence of the midregion of the vacA gene are reported. A total of 48 H. pylori strains isolated from gastric biopsy specimens obtained from 18 patients with peptic ulcer dyspepsia, 15 patients with nonulcer dyspepsia, and 15 asymptomatic H. pylori-infected subjects were studied. Highly heterogeneous restriction patterns were obtained by digestion of PCR products with SauII, BglII, and HhaI, whereas HaeIII digestion resulted in a strictly homogeneous profile for H. pylori strains isolated from 14 of 18 (77.7%) patients with peptic ulcer dyspepsia, but a strictly homogeneous profile was found for strains from only 8 of 15 (53.3%) patients with nonulcer dyspepsia (P = 0.163) and 5 of 15 (33.3%) asymptomatic H. pylori-infected subjects (P = 0.014). A potentially important aspect of the results obtained is the clinical relevance, since a single restriction pattern seems to be able to identify the majority of H. pylori strains associated with peptic ulcer disease.

Adult↗

Detection of serum antibodies to CagA and VacA and of serum neutralizing activity for vacuolating cytotoxin in patients with Helicobacter pylori-induced gastritis.

Thirty patients with dyspepsia, with histological diagnosis of gastritis, and with endoscopic diagnosis of peptic ulcer disease (PUD) (n = 13) or nonulcer dyspepsia (NUD) (n = 17) were admitted to the study. Helicobacter pylori vacuolating cytotoxin-producing strains (Tox+) were isolated from 14 (46.7%) patients, whereas non-cytotoxin-producing (Tox-) H. pylori strains were isolated from the remaining patients. Of 30 patients studied, 20 (66.7%) had serum cytotoxin neutralizing activity in vitro. Fourteen patients with Tox+ H. pylori strains showed serum cytotoxin neutralizing activity and serum immunoglobulin G (IgG) and IgA antibodies reactive with both 87-kDa H. pylori vacuolating cytotoxin (VacA) and 128-kDa cytotoxin-associated gene product (CagA) by immunoblotting using native enriched preparations of VacA and CagA proteins from H. pylori culture supernatants as the antigens. A 94-kDa antigen cross-reacting with the 87-kDa VacA protein could be demonstrated in culture supernatant with immune sera from humans and animals. All patients (n = 10) lacking serum neutralizing activity were also negative for IgG or IgA against VacA antigen, whereas 6 of the 10 patients showed IgG serum antibody responses against CagA antigen. The prevalence of antibodies to VacA and CagA antigens was significantly (P < 0.001) higher in patients with gastritis (20 and 26 patients for VacA and CagA, respectively, of 30 patients) than in H. pylori culture-negative controls (0 of 27 for both VacA and CagA) and in randomly selected blood donors (17 and 21 for VacA and CagA, respectively, of 120 subjects). All patients with PUD had antibodies to CagA, whereas 13 of 17 (76.5%) patients with NUD had anti-CagA antibodies. Serum IgG antibodies to VacA were present in 9 (69.2%) patients with PUD of 13 patients and in 11 (64.7%) patients with NUD of 17 patients. Anti-CagA antibodies seemed to correlate better with PUD than anti-VacA antibodies.

Adult↗

Characterization of a new isolate of Chlamydia trachomatis which lacks the common plasmid and has properties of biovar trachoma.

A Chlamydia trachomatis urethral isolate, alpha/95, yielding pgp3-negative but otherwise normal inclusions by immunofluorescence also gave negative results when pCT-homologous DNA was searched by PCR and Southern blotting. omp-1 sequence analysis identified alpha/95 as a new genotype B variant. These findings confirm that pCT is not required for chlamydial growth in vitro.

Amino Acid Sequence↗

Characteristics of non-O1 Vibrio cholerae isolated from the effluents of a treatment plant.

We report the results of a study concerning the characteristics of 19 Non-O1 Vibrio cholerae strains isolated from the incoming sewage and the effluents of the treatment plant in Bologna (Italy). These strains were compared to those of a strain of Vibrio cholerae biotype El Tor. The behaviour of the Non-O1 Vibrios was seen to be quite similar to those of the El Tor biotype in all aspects studied and antigenic correlations were found by means of immunoblotting and cytotoxin production on VERO cells. Since these bacterial strains may be pathogenic in humans, we believe it useful to describe some of their characteristics.

Agglutination Tests↗

Differences in the envelope proteins of Chlamydia pneumoniae, Chlamydia trachomatis, and Chlamydia psittaci shown by two-dimensional gel electrophoresis.

Analysis by two-dimensional gel electrophoresis of the N-laurylsarkosinate(Sarkosyl)-insoluble envelope complexes of L-[35]S-cysteine-labeled elementary bodies of Chlamydia pneumoniae strain IOL-207, Chlamydia trachomatis serovar LGV2, D, and F, and Chlamydia psittaci strain 6BC showed differences in the molecular charges of chlamydial outer membrane proteins. The apparent isoelectric point (pI) of the major outer membrane protein of C. pneumoniae strain IOL-207 was 6.4, whereas the pI of the major outer membrane protein of the C. trachomatis and C. psittaci strains differed little from one another, ranging from 5.3 to 5.5. The 60-kDa cysteine-rich protein of C. pneumoniae was the only 60-kDa chlamydial protein with a pI value (5.9) more acidic than that of the corresponding major outer membrane protein. As a general rule, the charges of both the 60-kDa and the low-molecular-mass (12-15 kDa) cysteine-rich proteins were widely variable, depending on the strain. However, in each individual strain, the variation of the charge of the 60-kDa protein had a compensatory change in the low-molecular-mass cysteine-rich protein.

Bacterial Outer Membrane Proteins↗

Detection of serum antibodies against Chlamydia pneumoniae by in vitro neutralization and microimmunofluorescence assays.

Antibodies to Chlamydia pneumoniae (IOL-207) were studied by the microimmunofluorescence (MIF) assay and in vitro neutralization in serum samples from 230 healthy adults and 332 patients at risk of C. pneumoniae infection. In the MIF test, the prevalence of immunoglobulin G (IgG) antibody was 53.9% among healthy subjects and 40.7% in the patients. An MIF antibody titre of > or = 1 : 512, consistent with acute infection by C. pneumoniae was documented in 23 out of 332 patients, and in none of the healthy adults. C. pneumoniae complement-dependent neutralizing antibody was detected in 25.8% and 31.2%, respectively of MIF-positive sera from healthy subjects and patients. The neutralizing antibody detection rate was 52.2% among the 23 patients with MIF titres of > or = 1 : 512. Complement-independent neutralization was observed in only 5 sera from healthy subjects and in 3 sera from patients. The complement-dependent neutralizing ability of sera significantly (p < 0.001) correlated with MIF titres.

Adolescent↗

Value of serology (ELISA) for the diagnosis of Helicobacter pylori infection: evaluation in patients attending endoscopy and in those with fundic atrophic gastritis.

In the present study we assessed the diagnostic accuracy of four commercial IgG enzyme-linked immunosorbent assay (ELISA) kits (Autoplate, H.pylori-EIA-Well, Enzygnost, Helori-test) and evaluated the performance of these tests in patients with fundic atrophic gastritis. Serum antibodies to Helicobacter pylori were measured in 70 out-patients attending endoscopy for dyspepsia and 43 patients with non-autoimmune fundic atrophic gastritis. Using the cut-off values recommended by the manufacturers, and comparing serological findings with gastric biopsy results of dyspeptic out-patients attending endoscopy, the four kits showed a sensitivity and specificity, respectively, of 91% and 96%, for Autoplate, 67% and 100% for H.pylori-EIA-Well, 79% and 100% for Enzygnost, and 81% and 96% for Helori-test. Evaluation in patients with atrophic gastritis revealed a high prevalence of antibodies to Helicobacter pylori (84%) and it demonstrated that patients with and those without gastric colonization by this microorganism had a similar rate of seropositivity (76-84% vs 50-78%). In conclusion, our data demonstrate that: a) this assay is a reliable and valid method to detect gastric colonization by Helicobacter pylori; b) positive serum antibody associated with a negative detection of Helicobacter pylori in the gastric mucosa suggests mucosal atrophy; c) patients with fundic atrophic gastritis should be excluded from studies investigating the value of serology in diagnosing Helicobacter pylori infection.

Adult↗

An immunofluorescence assay in microwell plate (IFA) for detecting serum antibodies to Chlamydia pneumoniae.

An indirect immunofluorescence assay using a 96 microwell cell culture plate (IFA) was developed for the detection of antibodies to C. pneumoniae. The results obtained by IFA on 230 sera of healthy subjects were compared with those obtained with the microimmunofluorescence test. The correlation coefficient for IgG detection with the two methods was 0.96 showing good agreement.

Adult↗

Influence of centrifugation on the infectivity of Chlamydia pneumoniae IOL-207.

The influence of centrifugation on the infectivity of Chlamydia pneumoniae IOL-207 was compared with seven serovars of C. trachomatis biovar trachoma, the three serovars of C. trachomatis biovar lymphogranuloma and four C. psittaci strains. Prolonged centrifugation from 1 to 4 hours resulted in a 3-fold increase in the infectivity of C. pneumoniae and variable or no increase in the infectivity of the other Chlamydia strains studied. These findings indicate that a 4 hour centrifugation of C. pneumoniae is recommended to improve the isolation procedures of this organism in cell cultures.

Animals↗

Partial characterization of an 89-kDa highly immunoreactive protein from Chlamydia psittaci A/22 causing ovine abortion.

An 89-kDa immunogen from Chlamydia psittaci A/22 causing ovine abortion was partially characterized. The 89-kDa protein, localized on the outer membrane complex of chlamydiae, was synthesized relatively early in the developmental cycle. The protein contained cysteine but was not extensively cross-linked by disulfide bonds. Treatment with proteases apparently did not cleave the protein. The infectivity of strain A/22 was partially (60%) reduced by treatment of chlamydial elementary bodies with monoclonal antibody BS/89 specifically reacting with the 89-kDa antigen. Species-specific as well as strain-specific antigenic determinants were present on the 89-kDa protein.

Abortion, Veterinary↗

Amperometric monitoring of bacteria-induced milk acidity using a platinum disc microelectrode.

The acidity induced by the action of bacteria in milk samples was monitored amperometrically by using a platinum microelectrode. The measurements were performed directly on commercial packs of milk, stored at 32 degrees C, and were continued for 9-10 d after inoculation. The data were compared with those obtained by measuring the pH of the samples and the results are discussed on the basis of the metabolism of each bacterial species. The effects of the following bacteria were examined: Staphylococcus aureus, Bacillus cereus, Streptococcus faecalis, Bacillus subtilis, Aeromonas, and Corynebacterium.

Animals↗