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B Joly

Publications and source records attributed to B Joly.

At least 55 records · Page 3Linked to original sources
PubMed1992

R-plasmid-encoded adhesive factor in Klebsiella pneumoniae strains responsible for human nosocomial infections.

Klebsiella pneumoniae strains involved in hospital outbreaks of nosocomial infections, such as suppurative lesions, bacteremia, and septicemia, were resistant to multiple antibiotics including broad-spectrum cephalosporins. Epidemiologic investigations revealed that the reservoir for these K. pneumoniae strains was the gastrointestinal tracts of the patients. The study of the adherence ability of the strains reported here showed that these bacteria adhered to the microvilli of the Caco-2 cell line. This adhesion was mediated by a nonfimbrial protein with a molecular mass of 29,000 Da designated CF29K. Pretreatment of bacteria with antibodies raised against CF29K or Caco-2 cells with purified CF29K prevented the adhesion of K. pneumoniae strains to Caco-2 cells. CF29K immunologically cross-reacted with the CS31A surface protein of Escherichia coli strains involved in septicemia in calves. Genes encoding CF29K were located on a high-molecular-weight conjugative R plasmid, which transferred to E. coli K-12. Transconjugants expressed a large amount of CF29K protein and adhered to the brush border of Caco-2 cells. These findings show that K. pneumoniae strains were able to colonize the human intestinal tract through a plasmid-encoded 29,000-Da surface protein. Hybridization experiments indicated that the gene encoding resistance to broad-spectrum cephalosporins by the production of CAZ-1 enzyme and the gene encoding the adhesive property to intestinal cells were both located on a 20- to 22-kb EcoRI restriction DNA fragment. Genes encoding aerobactin and the ferric aerobactin receptor were also found on this R plasmid.

Bacterial Adhesion↗
PubMed1992

Expression of receptors for enterotoxigenic Escherichia coli during enterocytic differentiation of human polarized intestinal epithelial cells in culture.

To study the expression of human intestinal receptors for enterotoxigenic Escherichia coli (ETEC), the human polarized intestinal epithelial cell line Caco-2 in culture and several subpopulations of HT-29 cells in culture--parental (mainly undifferentiated) HT-29 cells (HT-29 Std), an enterocytelike subpopulation obtained by selection through glucose deprivation (HT-29 Glc-), and an enterocytelike subpopulation obtained by selection through glucose deprivation which maintains its differentiation characteristics when switched back to standard glucose-containing medium (HT-29 Glc-/+)--were used. Since Caco-2 spontaneously differentiated in culture under standard culture conditions (in the presence of glucose) and HT-29 cells were undifferentiated when cultured under standard conditions (HT-29 Std) and differentiated when grown in a glucose-free medium (HT-29 Glc-), we studied the expression of the receptors for colonization factor antigens (CFA) I, II, and III and the 2230 antigen of ETEC in relation to enterocytic differentiation. We provide evidence that expression of ETEC CFA receptors develops in parallel with other differentiation functions of the cultured cells. The expression of ETEC-specific brush border receptors was studied by indirect immunofluorescence using antibodies raised against purified ETEC CFA. No ETEC receptors were detected in HT-29 Std or short-term-cultured Caco-2 cells. However, among the population of HT-29 Std cells, 2 to 4% of the cells were found to bind ETEC, and these cells expressed positive carcinoembryonic antigen immunoreactivity. This indicated that among the population of undifferentiated HT-29 cells, clusters of differentiated cells were present. ETEC CFA receptors were expressed in the apical and basolateral domains of differentiated HT-29 cells, whereas in differentiated Caco-2 cells only apical expression was observed. Both in HT-29 cells (HT-29 Glc-/+) and in Caco-2 cells cultured under standard conditions, ETEC CFA receptors develop as a function of day in culture. This indicated that the expression of the ETEC CFA receptors was a growth-related event. Indeed, ETEC CFA receptors developed in step with the apical expression of differentiation-associated proteins.

Antigens, Bacterial↗
PubMed1992

Inhibition of Haemophilus influenzae adherence to buccal epithelial cells by cefuroxime.

We studied the effect of subinhibitory concentrations of cefuroxime on the capacity of Haemophilus influenzae to adhere to buccal epithelial cells (BEC). Two encapsulated strains (serotype b) and two nonencapsulated, nontypable strains were studied. All four strains adhered strongly to BEC, with indices (mean number of bacteria adhering to a single BEC) ranging from 19 to 48. Subinhibitory concentrations of cefuroxime (serial dilutions from MIC/2 to MIC/32) were added to cultures in tryptic soy broth and their effect on adherence was tested after 18 h incubation at 37 degrees C. Adherence was diminished by more than 50% by concentrations of cefuroxime ranging from MIC/2 to MIC/8 and varied according to the strain studied. These results show that the adherence of H. influenzae to BEC is inhibited by subinhibitory concentrations of cefuroxime.

Adult↗
PubMed1991

Sequence and molecular characterization of the ROB-1 beta-lactamase gene from Pasteurella haemolytica.

The ROB-1 beta-lactamase-encoding plasmids from eight Pasteurella and two Haemophilus strains were compared by restriction endonuclease and hybridization analyses. Two types of ROB-1-encoding plasmids, which differed in size, were detected. One (4.1 kb) was found only in Pasteurella strains. The other (4.4 kb) was found in both Haemophilus influenzae and in one of the eight Pasteurella strains examined. These two plasmids shared multiple homologous fragments, suggesting that one was derived from the other. The ROB-1-encoding gene from Pasteurella haemolytica LNPB 51 was cloned and sequenced. An open reading frame of 915 nucleotides was found; it encoded a 305-amino-acid protein. Analysis of this amino acid sequence confirmed that the enzyme was found; it encoded a 305-amino-acid protein. Analysis of this amino acid sequence confirmed that the enzyme is a class A beta-lactamase. It had 32 to 48% homology with other class A enzymes and exhibited several common features of the gram-positive beta-lactamases. The ROB-1 mature protein, however, contained only one cysteine residue at position 123. These results suggest that ROB-1 is a link between beta-lactamases of gram-negative and gram-positive bacteria. An internal 230-bp DraI fragment from ROB-1 hybridized only with plasmid DNA from ROB-1-producing strains. This specific probe could be useful in epidemiological studies.

Amino Acid Sequence↗
PubMed1991

Scanning and transmission electron microscopic study of adherence of Escherichia coli O103 enteropathogenic and/or enterohemorrhagic strain GV in enteric infection in rabbits.

The GV strain (serotype O103:H2:K-), originally isolated from a diarrheic rabbit, is an enteropathogenic Escherichia coli strain that produces diarrhea without synthesizing the classical enterotoxins and that is not invasive. This strain is characterized by a 117-kb plasmid (pREC-1). Histological study of the gut by scanning electron microscopy and transmission electron microscopy was performed on the GV strain, on a derivative strain cured of pREC-1, and on transconjugants obtained by transfer of pREC-1 to nonpathogenic strains E. coli K-12 and 6100, not belonging to the O103 serogroup. The GV strain adhered to the epithelial cells of the ileum and large intestine, whereas the cured GV strain did not. Transfer of plasmid pREC-1 to E. coli K-12 or 6100 allowed the bacteria to attach to the intestinal mucosa in the same manner as that of the wild-type GV strain. Thus, pREC-1 seems to play an important role in attachment to and colonization of the intestinal tract of rabbits by E. coli serogroup O103. Scanning electron microscopy showed numerous bacteria attached together and closely associated with intestinal villi. Transmission electron microscopy revealed effacing lesions characteristic of enteropathogenic E. coli strains: effacing of microvilli and cuplike projections (pedestal formations) associated with an acute inflammatory and hemorrhagic response. In contrast with the results reported for rabbit pathogenic O15 strains, it appeared that the Peyer's patches were not involved in the early stages of infection with the O103 GV strain. This strain may represent a model for the study of the virulence and pathogenic effects of enteropathogenic and enterohemorrhagic E. coli strains.

Animals↗
PubMed1991

New adhesive factor (antigen 8786) on a human enterotoxigenic Escherichia coli O117:H4 strain isolated in Africa.

An enterotoxigenic Escherichia coli strain, E. coli 8786, of serotype O117:H4 produced only heat-stable enterotoxin and gave mannose-resistant hemagglutination with human and bovine erythrocytes. The strain adhered to the brush border of human enterocytes and to enterocytelike cell line Caco-2. Adhesion inhibition assays using Caco-2 cells with different adhesive factor extracts showed that the adhesive factor of E. coli 8786 is different from colonization factor antigen I (CFA/I). CFA/II, CFA/III of Darfeuille et al. (A. Darfeuille, B. Lafeuille, B. Joly, and R. Cluzel, Ann. Microbiol. Inst. Pasteur 134A:53-64, 1983), CS6, and antigen 2230. A bacterial surface protein, designated antigen 8786, with a molecular mass of 16,300 Da was responsible for the adhesion to intestinal cells. It was immunologically different from previously described adhesive factors as determined by immunoblotting. Antigen 8786 was detected on the bacterial cell surface and appeared to be nonfimbrial. NH2-terminal analysis of antigen 8786 showed no homology with the previously described adhesive factors. Nevertheless, antigen 8786 is closely related to the NH2-terminal sequence of Salmonella enteritidis fimbrin. A hybridization experiment using a synthetic oligonucleotide probe based on the NH2-terminal amino acid sequence of antigen 8786 revealed that the coding region was located on a 70-MDa plasmid.

Amino Acid Sequence↗
PubMed1991

R plasmid in Escherichia coli O103 coding for colonization of the rabbit intestinal tract.

One rabbit pathogenic Escherichia coli strain, belonging to serogroup O103, harbors a self-transferable 117-kb plasmid (pREC-1) encoding resistance to several antibiotics. The role of this R plasmid in the colonization of the digestive tract in specific-pathogen-free (E. coli O103-free) rabbits was studied. Five-week-old rabbits were inoculated with the wild-type strain, with its variant cured of the plasmid, with an E. coli K-12 strain, or with an untypeable E. coli strain from a healthy rabbit. No symptoms and no mortality were observed in animals inoculated with strains without the plasmid pREC-1, but 87.5% of the rabbits infected by the wild strain died, generally with bloody diarrhea, between days 5 and 15 postinfection. The weight gain of animals was strongly reduced. Transfer of the plasmid to the cured strain or to nonvirulent strains led these strains to induce the same pathology but with a lower mortality. Colonization of the gut by the O103 strain and symptoms of bloody diarrhea are thus related to the presence of the pREC-1 plasmid. The GV strain, which does not produce classical heat-labile enterotoxin or heat-stable enterotoxin and is not invasive, could be considered an enteropathogenic E. coli-like strain. The presence of a conjugative plasmid such as pREC-1 encoding both antibiotic resistance and virulence determinants in O103 E. coli from rabbits could represent a prominent epidemiological hazard under selective pressure by antibiotic therapy.

Animals↗
PubMed1991

Adhesive properties of Haemophilus influenzae to different human cells.

The adhesion of 19 nontypable strains and 3 typable (type b) Haemophilus influenzae to human cells was examined using buccal epithelial cells (BEC), the continuous HEp-2 cell line and human 0 erythrocytes. The strains were classified into three phenotypes, according to their adhesive properties. Phenotype 1 consists of strains that adhere to both buccal epithelial cells and HEp-2 cells. Phenotype 2 consists of strains that adhere to both buccal epithelial cells and erythrocytes and strains belonging to phenotype 3 adhere to none of the three cell types used. Among 22 strains studied, 18 (81.8%) belonged to phenotype 1, 2 (9.1%) to phenotype 2 and 2 (9.1%) to phenotype 3. Fimbriae were observed for 11 (61%) among the 18 adherent strains belonging to phenotype 1. The 7 nonpiliated strains adhered with a significant adhesion index, thus this results would indicate that a non fimbrial adhesin exists.

Bacterial Adhesion↗
PubMed1991

[Factors intervening in the variations of in vitro adhesion power of enterotoxinogenic colibacillus (ETEC) to human enterocytes].

Human enterotoxigenic Escherichia coli adhere to the brush border of human enterocytes. The mean number of bacteria adhering to one enterocyte (adhesion index) varied from 0.5 to 3.1 when the strains produce adhesins. Different factors related to enterocytes and to bacteria are involved in this variability. The number of bacteria which adhered to enterocytes issued from the same donor varied from from 0 to 12. Moreover the proportion of enterocytes on which several bacteria sticked did not exceed 20%. This variability might be due to the disparity in the maturation of the enterocytes. On the other hand, whatever the adhesion factors considered, the adhesion index varied according to the donors. ETEC strains did not express adhesion when bacteria were grown in a liquid medium but this capacity could be restored after transfer on solid medium. This phenomenon seemed like a phase-variation and appeared to be linked to a 4 to 6 kilobases (kb) plasmid. On the other hand, when the bacteria were grown on agar medium (CFA-agar or Mueller-Hinton agar) two phenotypes of colonies could be observed: large colonies (LC) which were composed of non-adhesive bacteria and small colonies (SC) which were composed of a majority of adhesive bacteria; when the number of subcultures was not too great, a majority of colonies presented the small colonies phenotype. The plasmid content analysis showed the segregation of a high molecular weight plasmid DNA (approximately 100 kb) for the bacteria issued from large colonies phenotype.

Age Factors↗
PubMed1990

Adhesion of enterotoxigenic Escherichia coli to the human colon carcinoma cell line Caco-2 in culture.

Enterotoxigenic Escherichia coli (ETEC) strains possessing colonization factor antigen I (CFA/I), CFA/II, CFA/III, and antigen 2230 were tested for their ability to adhere to the following cell lines: HeLa, HEp-2, HRT 18, Hutu 80, MDBK, MDCK, Vero, and Caco-2. ETEC strains adhered only to the Caco-2 cell line. Irrespective of the known adhesive factors, the ETEC strains that adhered to the brush border of human enterocytes also adhered to the Caco-2 cell line. The negative variants, which were cured of the plasmid encoding the adhesive factor, did not adhere. Adhesion of ETEC strains no longer occurred when the Caco-2 cells were pretreated with the homologous colonization factor antigen or when the bacterial cells were pretreated with homologous antibodies raised against the adhesive factors. This indicates that this adhesion is specific and that a different receptor exists for each type of adhesion factor. Electron micrographs of cross sections of the monolayer showed that the adhesion of ETEC strains to the brush border microvilli does not induce any lesion. Therefore, the Caco-2 cell line behaves in the same way as human enterocytes do.

Antigens, Bacterial↗
PubMed1990

[Piperacillin-amikacin combinations: killing curves].

Bactericidal activity as a function of time of piperacillin (PIP) and amikacin (AKN) alone and in combination was evaluated by killing curves technique on 23 clinical isolates: E. coli (6), K. pneumoniae (5), E. cloacae (6) and P. aeruginosa (6), for which the minimal inhibitory concentrations ranges of piperacillin were 0.25 to 64 mg/l and of amikacin 1 to 8 mg/l. For each species, the strains were chosen according to the most frequent phenotypes: beta-lactams susceptible, penicillinase (Pase), cephalosporinase (Case) and Pase + Case producers. Killing curves were carried out with the following concentrations (mg/l): piperacillin (2, 16, 64); amikacin (4, 8, 16); piperacillin (2) + amikacin (4); piperacillin (16) + amikacin (8); piperacillin (64) + amikacin (16). Antibiotic concentrations corresponded to pharmacokinetics and/or to critical values of piperacillin and amikacin. Bactericidal activity was defined as a 4 log 10 decrease in CFU/ml between 2 and 24 hours. When piperacillin (64) was combined with amikacin (16), the bactericidal effects were nearly the same as those with amikacin alone. But piperacillin (16) + amikacin (8) combination had bactericidal effect for the majority of strains (21/23) and it prevented for some of them the bacterial regrowth observed with amikacin alone at the same concentration. A bactericidal activity without regrowth (until the 24th hour) was obtained for 9 strains; 2 susceptible E. coli, 3 K. pneumoniae (chromosomal Pase producer) and 4 cefotaxime susceptible E. cloacae, with low dose combination piperacillin (2) + amikacin (4). Finally, only combinations piperacillin (64) + amikacin (16) or piperacillin (16) + amikacin (8) had bactericidal activity on 2 Ticarcillin-resistant P. aeruginosa, the two antibiotics being separatedly bacteriostatic.(ABSTRACT TRUNCATED AT 250 WORDS)

Amikacin↗
PubMed1990

[In vitro antibacterial activity of rokitamycin, a new macrolide antibiotic. Results of a multicenter study].

Minimal inhibitory concentrations (MIC) of rokitamycin (R) were evaluated by agar dilution for 914 bacterial strain isolated in 4 hospitals and classed as a function of susceptibility and resistance to Macrolides-Lincosamides-Streptogramines group (MLS). MICs of R ranged from 0.06 to 1 microgram/ml (mode MIC 0.25-0.5) on Staphylococci susceptible to MLS and on MLSB inducible strains; R was inactive on MLSB constitutive strains. MICs of R ranged from 0.008 to 0.5 microgram/ml (mode MIC 0.06 to 0.25) for Streptococci and Pneumococci susceptible to erythromycin (E) and from 0.06 to greater than 128 for strains resistant to E. Enterococci susceptible to E were inhibited by 0.06 to 0.5 microgram/ml (mode MIC 0.5) and strains resistant to E by 0.25 to greater than 128. Haemophilus were inhibited by 0.5 to 0.65 microgram/ml (mode MICs of R ranged generally from 0.016 to 0.5 microgram/ml (mode MIC 0.12) for C. perfringens and from 0.016 to 1 (mode MIC 0.06) for B. fragilis. Thus, R was shown to be among macrolide antibiotics of resistance strains. Its activity was superior to that of other products of this group spiramycin, josamycin, miokamycin, particularly on Gram positive cocci. R had a good activity on Neisseria, Branhamella, anaerobes and, as others macrolides, was poorly active on Haemophilus.

Anti-Bacterial Agents↗
PubMed1989

Adhesive properties of enteropathogenic Escherichia coli isolated from infants with acute diarrhea in Africa.

The adhesive properties of 69 enteropathogenic Escherichia coli (EPEC) strains were studied. The strains were isolated from diarrheal stools of infants in Burundi, Africa, and identified by serotyping with 12 classical EPEC O serogroup antisera. A test for adhesion to HEp-2 cells revealed that 52% of the strains showed localized adherence and 13% diffuse adherence. Localized adherence phenotype was found in previously described serogroups O86, O111, O125, O127, O128 and O142; strains belonging to another serogroup, O126, also exhibited localized adherence. Use of an EPEC adherence factor DNA probe in colony hybridization and in Southern blot techniques revealed that all strains exhibiting localized adherence and no strain exhibiting diffuse adherence produced a positive reaction; the genes were localized on high molecular weight plasmids (50-70 megadaltons). In vitro adhesion tests with human enterocytes performed concurrently with all 69 strains showed that only six of them adhered. These strains belonged to the O26, O117, O125, O128 and O142 serogroups. The adhesin CFA/I was detected only in the O128 Escherichia coli. The strain of serogroup O142 exhibited both adhesion to human enterocytes and localized adherence to HEp-2 cells, which suggests that the adhesive systems of enterotoxigenic and enteropathogenic Escherichia coli may coexist.

Bacterial Adhesion↗
PubMed1989

[Inhibition by nifurzide of the adhesion capacity of enteropathogenic colibacillus to HEP-2 cells].

Enteropathogenic Escherichia coli (EPEC) adhere in vivo to enterocytes. This adhesion capacity can be obtained in vitro with Hep-2 cells on which a characteristic localized adherence (LA) is observed. We studied the effect of subinhibitory concentrations (SIC) of nifurzide, a nitrofurane derivative, on this bacterial adherence phenomenon. Three EPEC strains are used: 11201 (026 serogroup), 7958 (0128) and 7836 (0142). Various SIC (MIC/2; MIC/4;...; MIC/32) were added either to the culture medium of bacteria or to the Eagle medium in which bacteria and Hep-2 cells were mixed during the adhesion experiments. In each case an adhesion index is determined. Nifurzide strongly inhibits the adherence capacity of the three strains when concentrations ranging from MIC/2 to MIC/16 where added in both culture media. On the other hand three other nitrofurane derivatives which have no antibacterial effect did not inhibit adherence. The mechanism of the adherence inhibition by nifurzide is discussed.

Anti-Bacterial Agents↗
PubMed1988

Characterization of Haemophilus influenzae isolated from patients with otitis media.

Two hundred and twenty-three Haemophilus influenzae strains isolated from patients with otitis media throughout France were characterized by biotype, serotype, antibiotic susceptibility, type of beta-lactamase production and human erythrocyte agglutination properties. All strains fell in one of two groups. One group consisted of encapsulated type b strains, 50% of which were biotype I, often resistant to ampicillin (38.5% of beta-lactamase producing strains) and seldom positive for haemagglutination (3.8%). The second group was composed of non-encapsulated strains, 42% of which were of biotype II, 10.6% beta-lactamase producers and 10.5% positive for haemagglutination.

Adolescent↗