PubMed Health⌕ Search

Biomedical subjects

L G Mathieu

Publications and source records attributed to L G Mathieu.

At least 19 recordsLinked to original sources

Evolution of the genomic systems of prokaryotes and its momentous consequences.

The earliest self-reproducing cell on Earth, our common ancestor, was probably as small as present-day bacteria. It gave rise to a very large and durable clone whose descendants must have been the only living occupants of the oceans for about one thousand million years. They reached astronomical numbers of separate, disjunct cells, and synthesized many new genes. Their small volume could not accommodate ever larger genomes and useful new genes replaced resident, less successful sequences, thus increasing diversity and the number of strains with highly specialized, distinct, bioenergetic potentialities. Also, selective pressure favored strains able to participate successfully in division of labor and in the sharing of diverse abilities in mixed communities, counterbalancing the limited capacities of individual genomes. Lateral gene transfer mechanisms appeared and were progressively improved, furthering the development of diversity. The prokaryotes' constructive evolution resulted in the formation of a worldwide web of genetic information, and a global bacterial superbiosystem (superorganism). By contrast, eukaryotic evolution of organisms has been typically Darwinian. Diversification of eukaryotic organisms was, however, considerably enriched and accelerated by symbioses with prokaryotes. The more broadly diversified bioenergetic potential of prokaryotes considerably increased the diversity of eukaryotes. Without their participation, our biosphere would have remained much less diverse and less dynamic. Environmental homeostasis has been maintained all along by guided bacterial evolution.

Evolution, Molecular↗

A powerful bacterial world.

Bacteria (prokaryotes) were the sole form of life on earth for some two billion years--roughly half its history. During this time they evolved into a giant, global superorganism and developed a remarkable mechanism for the creation and exchange of genetic material. Apart from its intrinsic interest, this has practical significance, for example in the development of multiple resistance to antibiotics of pathogenic bacteria such as those of tuberculosis. Eukaryotes, with nucleated cells, may have developed from a permanent symbiosis of three or more prokaryotes.

Bacteria↗

Gastrointestinal colonization and systemic dissemination by Candida albicans and Candida tropicalis in intact and immunocompromised mice.

Gastrointestinal colonization and systemic dissemination by Candida albicans and Candida tropicalis were compared in intact and immunocompromised mice. Five-day-old CFW mice were inoculated by the oral-intragastric route with 1.0 x 10(7) CFU of two C. albicans and two C. tropicalis strains isolated from the blood of patients with acute leukemia and with C. albicans 4918 and its cerulenin-resistant mutant 4918-10. C. albicans and C. tropicalis spread to the lungs, liver, and kidneys within 30 min postinoculation, and organ CFU of the two species were comparable over the following 10 days. Close association of blastoconidia with the villous surface of the small intestine resulted in lysis of microvilli and then progressive invasion of villi. Blastoconidia within villi were surrounded by a conspicuous zone of clearing. Persistent colonization of the small and large intestines by C. albicans blood isolates and strains 4918 and 4918-10 was similar for 31 days after inoculation, but consistently exceeded that of C. tropicalis. In mice colonized with C. albicans, immunosuppression with cortisone acetate and cyclophosphamide on days 30 and 33 after inoculation increased stomach CFU 40- to 370-fold and intestinal CFU 30- to 80-fold. In contrast, persistent colonization by C. tropicalis was undetectable before immunosuppression and only became apparent after treatment. C. albicans disseminated more frequently and with higher organ CFU than C. tropicalis. Despite this fact, 20% of mice infected with C. tropicalis died, compared with 4% infected with C. albicans blood isolates. Indirect immunofluorescence revealed penetrative growth by Candida hyphae exclusively in the mucosa and submucosa of the stomach from immunosuppressed, persistently colonized mice. Taken together, the data indicate that C. tropicalis appears to be more virulent than C. albicans and that factors responsible for gastrointestinal colonization, systemic dissemination, and mortality in immunocompromised mice may not be identical.

Animals↗

Adherence of gram-positive and gram-negative bacterial strains to human lung fibroblasts in vitro.

The adherence to eukaryotic cells of Escherichia coli, Neisseria gonorrhoeae, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis and the yeast Candida albicans was studied by light microscopy with an in vitro micromethod involving different cell lines. The method is inexpensive, consumes little time and material, and is reproducible. It was used to show that the gram-positive Cowan I strain of S. aureus, which naturally forms protein A on its surface, adheres in much larger numbers to human lung fibroblasts than the protein A-free Wood 46 strain, the strain of S. epidermidis, and the encapsulated Smith strain. The presence of a capsule on the latter strain apparently prevented its attachment to the fibroblasts. Among the gram-negative species studied, a piliated clinical isolate of N. gonorrhoeae, displaying the opaque colonial phenotype, adhered in larger numbers than another isolate lacking pili and displaying the transparent phenotype. E. coli K12 attached slightly to the cell line, whereas P. aeruginosa adhered to it moderately. One strain of C. albicans tested did not attach in any detectable numbers. No clear correlation between bacterial cell surface hydrophobicity, as evaluated by the hexadecane assay, and adherence to eukaryotic cells could be demonstrated for these microorganisms. With our method, bacterial attachment proceeded best at 37 degrees C and did not require more than 1 h of contact with the cell monolayer. The method described revealed differences in the adherence to eukaryotic cells, not only among species, but also between strains of the same species.

Candida albicans↗

Inhibitory effects of mannose and N-acetylneuraminic acid on the envelope and the adherence of a Neisseria gonorrhoeae isolate to rabbit mesentery.

Sugars and sugar derivatives were investigated as potential inhibitors of adherence of Neisseria gonorrhoeae; in this experiment rabbit mesentery was used as a reliable and convenient tissue. The adherence of N. gonorrhoeae isolate 2299 was reduced by half and more, as compared to controls, in the presence of 1,0 mg N-acetylneuraminic acid or D-mannose per ml of culture media, but not by the other sugars tested. Our data suggest that the N. gonorrhoeae isolate used in this work has mannose-binding related activity which participates in the process of its adherence to rabbit mesentery cells. In addition we observed that pretreatment of rabbit mesentery with concanavalin A, but not with peanut and wheat germ agglutinins, markedly decreased the adherence of gonococci as determined by microscopic counts of adhering bacterial cells.

Adhesiveness↗

Effect of subminimal inhibitory concentrations of antibiotics on adherence of Neisseria gonorrhoeae in an experimental model.

The effect of subminimal inhibitory concentrations (subMICs) of six commonly used antibiotics on adherence of Neisseria gonorrhoeae to rabbit mesentery was studied. We found that, at 1/2 MIC, ampicillin, cefoxitin and penicillin G decreased adherence of gonococci by more than 65%, and erythromycin by approximately 40%; doxycycline and tetracycline had no appreciable effect. Results suggest that subMICs of some antibiotics could affect interactions of gonococci with some mammalian cells, more specifically adherence, a critical step in the pathogenesis of an infection.

Adhesiveness↗

Elimination of plasmidic resistance to ampicillin and of enterotoxinogenicity in certain enteric gram-negative bacteria after incubation with high concentrations of amprolium.

We have observed that treatment with high concentrations of a thiamine analog (amprolium) can lead to the elimination of the plasmidic resistance to ampicillin and the production of enterotoxin in wild Escherichia coli strains and in E coli and Salmonella typhimurium strains which had received the pKM101 plasmid through bacterial conjugation. By computer analysis, we also have determined that there is a highly significant (P less than 0.01) synergism between ampicillin and amprolium which reduces considerably the growth of certain enteric bacterial strains which have a plasmidic resistance to ampicillin and which were not markedly affected by amprolium alone, in our experimental conditions. Our data indicate that the rate of loss of the plasmid pKM101 after treatment of R+ bacterial strains with amprolium can be increased.

Ampicillin↗

Analysis of interactions between treatments applied concurrently onto biological systems.

A program, written in BASIC, has been developed to analyse results of experiments studying individual and combined effect of two treatments (physical or chemical) on a biological system. Analysis is done in regard to: (1) level of significance selected for statistical test; (2) number of data available; (3) expected combined effect of treatments resulting from addition which is computed using an original formula; (4) experimental hypothesis tested. Using this program we have determined that there is a highly significant synergism between ampicillin (antibiotic) and amprolium (antiparasitic drug) to reduce the growth of some enteric bacterial strains which have a plasmidic resistance to ampicillin. Other applications are suggested.

Anti-Bacterial Agents↗

Antibiotic susceptibilities and auxotypes of Neisseria gonorrhoeae strains from women with pelvic inflammatory disease or uncomplicated infections.

The purpose of the present study was to compare the auxotypes and the minimal inhibitory concentrations of four antibiotics for 50 isolates from women with gonococcal pelvic inflammatory disease with those of 55 isolates from uncomplicated anogenital gonococcal infections. No significant differences in auxotype patterns and susceptibilities were found between isolates from the two groups.

Anti-Bacterial Agents↗

Modulation by staphylococci of the response of mice infected with Neisseria gonorrhoeae.

Most strains of Neisseria gonorrhoeae are rapidly eliminated from the peritoneal cavity of mice following an experimental infection. However, we could modify the rate of clearance of N. gonorrhoeae by preteating (48 h) the mice with sublethal doses of Staphylococcus aureus coagulase-positive strains which provoke the rapid appearance of activated phagocytes; we showed that these have increased activities of their B-galactosidase and their enzymes able to reduce nitroblue tetrazolium (NBT). Presumably, these activated phagocytes participated in the more efficient elimination of N. gonorrhoeae cells from the peritoneal cavity, that is 3 log units instead of 1 for the control, within the first 2 hours after their injection. There was no increase in the clearance of N. gonorrhoeae from the peritoneal cavity of mice when these were pretreated with cyclophosphamide. The use of enzyme-treated staphylococci (protoplasts) showed that some component of the coagulase-positive S. aureus cell wall seems responsible for the stimulation observed. This modulation of the response of mice to N. gonorrhoeae could serve as an improved method to evaluate the virulence of different N. gonorrhoeae strains.

Animals↗

In vitro adhesion of Staphylococcus strains to rabbit tissues.

Differences in the in vitro adhesion of Staphylococcus strains on rabbit tissues were evaluated by viable unit counts and radio-labeling. Among coagulase-positive strains, two freshly isolated strains and the Cowan I strain adhered much more than the Wood 46 strain especially to peritoneum. In addition, the Smith strain, which is coagulase-positive and encapsulated, showed on rabbit peritoneum a degree of adhesion smaller than that of the Cowan I strain and similar to that of the Wood 46 strain. These quantitative results were corroborated by examination of the bacteria on thin sections of peritoneum or on whole mesentery samples with fluorescence microscopy. With fluorochroming either with acridine orange or fluorescent homologous antibodies, we could detect a preferential localization of the bacteria on the epithelium and also on fibers of the mesentery. Even if the Cowan I strain produces Protein A, this substance does not seem to have been involved in the adhesion of this strain to rabbit tissues, since IgG did not affect the degree of its adhesion. Treatment by Na metaperiodate of the peritoneum had an inhibitory effect of about 50% on the degree of adhesion; several other known inhibitors had no effect. The higher affinity of the Cowan I strain for the peritoneum or the mesentery, when compared with the Wood 46 strain, is an observation which could contribute to the study and understanding of adhesion, presumably an element of the experimental pathogenicity of Staphylococcus.

Animals↗

Changes in the activity of antifungals in mixed cultures of bacteria and yeasts.

When Candida cells were grown in mixed cultures with Escherichia coli, Salmonella typhimurium, or Pseudomonas aeruginosa, their sensitivity to antifungal agents affecting sterol metabolism or functioning (polyenes and miconazole) was increased by several log units. This phenomenon was overcome by the addition to the culture medium of sugars utilized either by the bacteria or the yeast. Antifungals without direct effect on sterol metabolism (5-fluorocytosine, methylparaben, and 4-hydroxyquinazoline) were not more active in mixed than in pure cultures.

Amphotericin B↗

Increase in the in vitro susceptibility of Staphylococcus aureus to antimicrobial agents in the presence of Candida albicans.

In experiments with mixed cultures of Staphylococcus aureus and Candida albicans both in the absence and in the presence of 5-fluorocytosine (5-FC), we have observed that (1) there is an inhibition of S. aureus growth in mixed cultures with C. albicans in media supplemented with 1 microgram/mL of 5-fc and that 5-FC has no effect on staphylococci in pure cultures; (2) this inhibition occurred with clinically isolated and laboratory strains and could be reversed by specific metabolites; (3) Staphylococcus aureus was inhibited by filtrates of C. albicans cultures treated with 5-FC and this seemed to be favored by some C. albicans filterable product which can affect the cell wall and the permeability of the staphylococcal cells since they become sensitive to 5-FC; (4) nine other commonly used antimicrobials showed an increased inhibitory activity against S. aureus in mixed cultures with C. albicans; and (5) there is a decrease in the number of precipitating antigens of S. aureus and of the activity of alpha toxin when this species was grown with both C. albicans and 5-FC. Our results indicate that the susceptibility of some species to antimicrobials could be significantly modified in the presence of other species. One cannot exclude that a similar phenomenon could happen in hosts under treatment with antibiotics against infection.

Anti-Bacterial Agents↗

Increased in vitro sensitivity of Candida albicans to amphotericin B when grown in mixed culture with Escherichia coli.

The growth of Candida albicans was inhibited by some Escherichia coli strains both in conventional batch cultures and also in a chemostat under conditions of constant addition of fresh medium. Concentrations of 0.2 microgram amphotericin B per millilitre and of 2 microgram nystatin per millilitre, which caused a slight inhibition of C. albicans in pure culture, exerted a strong fungicidal effect when the yeast was placed in mixed cultures with certain strains of E. coli. Candida albicans cells, inhibited by either E. coli or in mixed culture with polyene antibiotics, appeared larger and less uniformly stained by acridine orange than control cells from pure cultures. Addition of chloramphenicol to the mixed cultures, in quantities sufficient to kill the E. coli cells, abolished the increased sensitivity of C. albicans to amphotericin B or nystatin. In preliminary in vivo tests, E. coli did not sensitize C. albicans to the polyene antibiotics.

Amphotericin B↗

[Diminution of the antibacterial activity of antibiotics in cultures and in experimental mixed infections].

We have studied interactions between Staphylococcus aureus and Pseudomonas aeruginosa in the absence and the presence of four different antimicrobials in mixed cultures and experimental infections. These two bacterial species, in addition to having different properties, are known to be opportunistic pathogens often present in human microflora. Two main aspects have been investigated and they are related to modifications in two species affecting their equilibrium in the mixed bacterial population and also their pathogenicity markers. Our results indicate that individual growth of S. aureus and P. aeruginosa is not modified in vitro in mixed cultures in the absence of antimicrobials; in vivo, in mouse peritoneal cavity, there is a synergism favorable to S. aureus. In the presence of rifamycin SV and three cell wall inhibitors, pencillin G,D-cycloserine, and vancomycin, we have observed that P. aeruginosa protected S. aureus against the inhibitory effect of these antimicrobials in vitro and in vivo. Such results were obtained in different conditions of culture, stationary, shaken, and in special apparatuses, an "Ecologen" and a "Chemostat." When any one of the antimicrobials was allowed to be in contact for 6 to 8 h with P. aeruginosa cells in a culture, we observed a decrease in their inhibitory effects against S. aureus. These results were supported by microscopical observation. It seems that the inhibitory effects of the antimicrobials have hindered the formation of toxic products of S. aureus, e.g., alpha toxin, and that it was not restored in the presence of P. aeruginosa. Conversely, P. aeruginosa remained apparently unchanged through all these experiments. Our observations may imply that the inhibitory effect of an antimicrobial towards a bacterial species may be significantly decreased in the presence of another species, sometimes present in human microflora.

Animals↗

Production of colicin V in vitro and in vivo and observations on its effects in experimental animals.

In recent years, a possible relationship between pathogenicity and colicinogeny in some Escherichia coli strains responsible for gastrointestinal infection and bacteremia in man and animals has been inferred. Using enterotoxigen-negative, colicin V-producing E. coli strains, we have (i) elaborated a simple in vitro method for producing greater yields of colicin V free of bacterial cells and large, non-dialyzable molecules; (ii) detected the presence of the bacteriocin in peritoneal fluids of moribund mice injected intraperitoneally 18 h previously with colicin V-producing strains (in these mice, Col V+ exconjugants survived and multiplied more extensively than the Col V- recipient strains from which they were derived; and (iii) observed an increased vascular permeability and inflammatory response in rabbits and guinea pigs when a culture supernatant demonstrating colicin activity was injected intradermally. The vascular response obtained after the injection of either a colicin V-containing dialysate alone or that of a trypsinized colicin-containing supernatant was always smaller than when the colicin V-active supernatant was injected. An enterotoxin-free dialysate containing colicin V also increased markedly in rabbits and guinea pigs the mild inflammatory reaction that occurred in the skin when purified endotoxin was injected subcutaneously in microgram doses. Our results are consistent with the hypothesis that colicin V may act as a virulence factor in some E. coli strains.

Animals↗

[Production of colicins V and V2 in vitro and in vivo. Study of their inhibitory action on phagocytosis by peritoneal macrophages].

In recent years, a possible relationship between pathogenicity and colicinogeny in some Escherichia coli colicin V-producing strains had been inferred. In our laboratory, we have elaborated a simple in vitro method for the production of colicin V free of large, non dialyzable macromolecules and presumably of bacterial endotoxin. This allows study of the effects of colicin V in vivo without an undesirable added physiological response of the experimental animal to endotoxin. All the Col V+ strains we have studied displayed a greater ability to survive in the peritoneal cavity of mice than the Col V- strains. Also, we have detected colicin V in peritoneal fluids of agonizing mice injected with Col V+ strains. Phagocytosis by peritoneal macrophages seemed to be inhibited in vitro in the presence of colicin V. Colicin V is not toxic in vivo in low concentration, after intraperitoneal or intravenous injection but it may favor the multiplication and the invasiveness of the strains that produce it.

Animals↗