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Biomedical subjects

J M Moreau

Publications and source records attributed to J M Moreau.

At least 19 recordsLinked to original sources

Assessing the contamination potential of freshly extracted Escherichia coli biofilm cells by impedancemetry.

Planktonic bacteria passing to a sessile state during the formation of a biofilm undergo many gene expression and phenotypic changes. These transformations require a significant time to establish. Inversely, cells extracted from a biofilm should also require a significant time before acquiring the same physiological characteristics as planktonic cells. Relatively few studies have addressed the kinetics of this inverse transformation process. We tested one aspect, namely, the contamination potential of freshly extracted Escherichia coli biofilm cells, precultured in a synthetic medium, in a rich liquid growth medium. We compared the time between inoculation and the beginning of the growth phase of freshly extracted biofilm cells, and suspended exponential and suspended stationary phase cells precultured in the same synthetic medium. Unexpectedly, the lag time for the extracted biofilm cells was the same as the lag time of the suspended exponential phase cells and significantly less than the lag time of the suspended stationary phase cells. The lag times were determined by an impedance technique. Cells extracted from biofilms, i.e., biofilms formed in canalizations and broken up by hydrodynamic forces, are an important source of contamination. Our work shows, in the case of E. coli, the high potential of freshly extracted biofilm cells to reinfect a new medium.

Bacterial Adhesion↗

[Autosomal dominant syndrome of retinal arterial tortuosity].

INTRODUCTION: Autosomal dominant syndrome of retinal arterial tortuosity is a rare condition, often discovered after a benign macular hemorrhage. CASE REPORT: We report here the case of a 52-year-old man who was refereed to our center for an abrupt decrease in vision after an effort. The initial visual acuity was 2/10 for distance and Parinaud 6 for near vision. The biomicroscopic examination showed a small foveal hemorrhage associated with loops and bilateral vascular tortuosities limited to the arterioles. The aspect evoked inherited retinal arteriolar tortuosity. Questioning the patient revealed an antecedent of macular hemorrhage in the patient's sister that had spontaneously resolved. After a few months, redfree photographs were obtained from the two asymptomatic daughters of the patient, which showed a dominant arterial tortuosity in one of the two daughters, confirming the familial aspect of the disease. CONCLUSION: The case described here illustrates the advantage of biomicroscopy in establishing the diagnosis and the usefulness of questioning the patient further to disclose family history. Imagery studies complement the examination to eliminate other causes for the decrease in vision. Some recently published data suggest an advantage to including at least the search for a microscopic or macroscopic hematuria during the assessment.

Genes, Dominant↗

A radio frequency electric current enhances antibiotic efficacy against bacterial biofilms.

Bacterial biofilms are notably resistant to antibiotic prophylaxis. The concentration of antibiotic necessary to significantly reduce the number of bacteria in the biofilm matrix can be several hundred times the MIC for the same bacteria in a planktonic phase. It has been observed that the addition of a weak continuous direct electric current to the liquid surrounding the biofilm can dramatically increase the efficacy of the antibiotic. This phenomenon, known as the bioelectric effect, has only been partially elucidated, and it is not certain that the electrical parameters are optimal. We confirm here the bioelectric effect for Escherichia coli biofilms treated with gentamicin and with oxytetracycline, and we report a new bioelectric effect with a radio frequency alternating electric current (10 MHz) instead of the usual direct current. None of the proposed explanations (transport of ions within the biofilm, production of additional biocides by electrolysis, etc.) of the direct current bioelectric effect are applicable to the radio frequency bioelectric effect. We suggest that this new phenomenon may be due to a specific action of the radio frequency electromagnetic field upon the polar parts of the molecules forming the biofilm matrix.

Anti-Bacterial Agents↗

First detection of resistance to QoI fungicides in Mycosphaerella graminicola on winter wheat in Belgium.

A total of 740 Mycosphaerella graminicola strains were isolated between 2000 and 2002 from winter wheat F1 or F2 leaves showing Septoria leaf blotch lesions (SLB) collected mainly at the soft dough stage in fungicide trials, analysing at 12 locations in Belgium the possibilities and risks associated with the use of epoxiconazole and azoxystrobin at various doses, mixtures and application dates. Fungicide sensitivity tests were performed in microtitre plates on potato dextrose broth amended with various concentrations of azoxystrobin. A wide range of sensitivity to azoxystrobin was observed, with EC50 values ranging for 735 strains between 0.002 to 0.7 microg/ml, the highest frequency gradually shifting from EC50 classes 0.01 and 0.02 microg/ml azoxystrobin in 2000 to EC50 classes 0.02 and 0.04 microg/ml in 2002. No clear selection effect of particular fungicide use strategies was observed. Among the 382 strains isolated in 2002, five originating from 2 locations, showed azoxystrobin EC50 values >1 microg/ml. On medium amended with 100 microg/ml salicylhydroxamic acid (SHAM), 58% of the 2002 strains were strongly inhibited, which affected adequate azoxystrobin ED50 determination. This suggests widespread occurrence of M. graminicola strains relying in vitro on the alternative respiration pathway. In the presence of SHAM, strains 339 and 880 showed azoxystrobin EC50 values of 3 and >30 microg/ml, respectively. This high level of resistance to a QoI fungicide was confirmed by analysing mycelium growth inhibition on PDA. Cross-resistance to trifloxystrobin and kresoxim-methyl was demonstrated. Greenhouse assays on wheat plants revealed that control of QoI resistant strains by azoxystrobin is decreased, compared to control of sensitive ones. This highlights the risk of resistance to QoI fungicides also in M. graminicola populations, although up to now no decrease in field performance was noticed. It is recommended to delay build up of QoI resistance by an integrated approach, combining optimised fungicide use with the choice of SLB resistant cultivars and the application of farming practices promoting stubbles break down and so the reduction of the teleomorph stage.

Acrylates↗

Assessment of tumour response to chemotherapy for metastatic colorectal cancer: accuracy of the RECIST criteria.

Evaluation of tumour size modifications in response to treatment is a critical issue in the management of advanced malignancies. In 1981, the World Health Organization (WHO) established guidelines for tumour response assessment. These WHO1981 criteria were recently simplified in a revised version, named RECIST (Response Evaluation Criteria in Solid Tumours), which uses unidimensional instead of bidimensional measurements, a reduced number of measured lesions, withdrawal of the progression criteria based on isolated increase of a single lesion, and different shrinkage threshold for definitions of tumour response and progression. In order to validate these new guidelines, we have compared results obtained with both classifications in a prospective series of 91 patients receiving chemotherapy for metastatic colorectal cancer. Data from iterative tomographic measurements were fully recorded and reviewed by an expert panel. The overall response and progression rates according to the WHO1981 criteria were 19% and 58%, respectively. Using RECIST criteria, 16 patients were reclassified in a more favourable subgroup, the overall response rate being 28% and the progression rate 45% (non-weighted kappa concordance test 0.72). When isolated increase of a single measurable lesion is not taken into account for progression with the WHO1981 criteria, only 7 patients were reclassified and the kappa test was satisfying, i.e. > or =0.75, for the whole population as well as for each of the responding and progressive subgroups. Since it provides concordant results with a simplified method, the use of RECIST criteria is recommended for evaluation of treatment efficacy in clinical trials and routine practice.

Adenocarcinoma↗

Lessons from the year 2001 Mycosphaerella graminicola epidemic on winter wheat in Belgium.

Infection by Mycosphaerella graminicola (anamorph Septoria tritici) was monitored between April and July 2001 on F6 to flag leaf in 11 farmers' fields or fungicide trials. Data were analysed by mean of the decision support system "Proculture" which links an automatic weather station of the PAMESEB network to a particular field, simulates plant development with adjustment by one phenological observation during the stem elongation and analyses superposition of emerged leaves and infection events (http://www.fymy.ucl.ac.be/proculture). Several climatic events favourable for the infection and dissemination of M. graminicola occurred between October 2000 and March 2001 and allowed build up of a large amount of inoculum on the lower leaves at the end of the winter. The start of stem elongation was associated with frequent rainy periods during April, causing early infection of F5, F4 and up to F3 in some precocious fields. Dry weather with only a few local showers during most of May and June slowed down spread of infection to the upper leaves, leading to absence of M. graminicola infection of the flag leaf in 9 out of the 11 fields. Yield increase by a single fungicide spray ranged from 800 to 2200 kg/ha. A second treatment was cost effective in none of the fields. The interest and limitation of the decision support system for understanding M. graminicola epidemic and for guiding decision on spray timing are discussed.

Belgium↗

Pseudomonas aeruginosa protease IV enzyme assays and comparison to other Pseudomonas proteases.

Pseudomonas aeruginosa secretes multiple proteases that have been implicated as virulence factors and the detection of each specific enzyme can be difficult to determine. Unlike the three Pseudomonas enzymes that have been well characterized (elastase A, elastase B, and alkaline protease), the activity of protease IV in multiple assays has yet to be described. This study defines new assays for Pseudomonas proteases and compares protease IV activity to the activities of elastase A, elastase B, and alkaline protease. Six in vitro assays were studied: zymography, elastin congo red assay, staphylolytic assay, colorimetric peptide assay, solid-phase colorimetric peptide assay, and poly-l-lysine degradation. Casein zymography distinguished protease IV from elastase B and alkaline protease, and gelatin zymography differentiated all four proteases. The elastin congo red assay detected mainly elastase B while the staphylolytic assay was specific for elastase A. Protease IV activity was assayed specifically by the colorimetric assay and two new assays, the solid-phase colorimetric assay and degradation of poly-L-lysine in the presence of EDTA. Alkaline protease could be specifically assayed by poly-L-lysine degradation in the presence of N-alpha-p-tosyl-L-lysine chloromethyl ketone. The results identified three specific assays for protease IV, a new assay specific for alkaline protease, and showed that protease IV has a distinct enzymatic specificity relative to the three other Pseudomonas proteases.

Colorimetry↗

The effectiveness of tobramycin and Ocuflox in a prophylaxis model of Staphylococcus keratitis.

PURPOSE: To determine the effectiveness of prophylactic antibiotic treatment prior to intra-corneal infection with Staphylococcus aureus. METHODS: One topical drop of Tobrex (0.3% tobramycin), tobramycin (0.3%) in the Tobrex vehicle with 0.05% dodecyl maltoside (DDM)/4.0% hydroxypropylmethycellulose (HPMC), Ocuflox (0.3% ofloxacin) or DDM/HPMC vehicle were applied to rabbit eyes at one or five hours prior to injection of bacteria. Approximately 500 colony-forming units (CFU) of S. aureus strain 8325-4 were injected into the corneal stroma. Rabbits were sacrificed five hours after infection and corneal homogenates were cultured to determine the number of colony forming units (CFU) per cornea. RESULTS: Rabbits treated at five hours prior to infection with tobramycin-DDM/HPMC reduced the bacterial load by approximately 2.4 log CFU/cornea as compared to the untreated control (3.47 +/- 0.98 vs. 5.71 +/- 0.14 log CFU/cornea, respectively; P = 0.0010); however, Ocuflox, Tobrex, or DDM/HPMC vehicle did not significantly reduce the log CFU (P >or= 0.4837). Rabbits treated at 1 hour prior to infection with Ocuflox or tobramycin-DDM/HPMC had significantly reduced CFU/cornea (1.31 +/- 0.86 and 0.48 +/- 0.31 log CFU/cornea, respectively) as compared to the untreated group (5.71 +/- 0.14 log CFU/cornea; P or= 0.2312). CONCLUSIONS: This pre-treatment model of Staphylococcus keratitis quantitatively measured the prophylactic effectiveness of topical antibiotic formulations. An important finding was that a tobramycin-DDM/HPMC formulation was highly effective as a prophylactic medication.

Animals↗

Effectiveness of ciprofloxacin and ofloxacin in a prophylaxis model of Staphylococcus keratitis.

PURPOSE: To determine the effectiveness of prophylactic fluoroquinolone treatment against staphylococci in a rabbit keratitis model. METHODS: Prophylactic ciprofloxacin or ofloxacin was applied as one topical drop 15 minutes before infection or as one drop at three time points (19, 17, and 15 minutes) before infection. In a second experiment, rabbits were treated with two, three, or four drops of ciprofloxacin 1 hour before infection. Approximately 250 colony-forming units (CFUs) of Staphylococcus aureus were injected intrastromally, and CFUs were determined 5 hours after infection. RESULTS: The CFUs per cornea in all treatment groups were significantly less than the 5.6 +/- 0.11 log CFUs per cornea in the untreated group ( p < or = 0.0001). Rabbit eyes treated 15 minutes before infection with Ciloxan or Ocuflox had 0.96 +/- 0.48 log CFUs per cornea (three of six sterile corneas) or 1.26 +/- 0.31 log CFUs per cornea (one of six sterile corneas), respectively ( p = 0.5226). Eyes treated with Ciloxan 19, 17, and 15 minutes before infection had 0.0 +/- 0.0 log CFUs per cornea, and all eyes were sterile, whereas eyes treated with Ocuflox had 0.98 +/- 0.48 log CFUs per cornea and two of six eyes sterile ( p = 0.0435). Eyes treated 1 hour before infection with two, three, or four drops of Ciloxan had 2.61 +/- 0.69 log CFUs, 1.23 +/- 0.32 log CFUs, or 0.85 +/- 0.28 log CFUs per cornea, respectively, which was significantly less than untreated eyes ( p < or = 0.0001). CONCLUSIONS: Multiple topical drops of a fluoroquinolone administered prophylactically were effective for subsequent staphylococcal ocular infection.

Animals↗

Phospholipase A(2) in rabbit tears: a host defense against Staphylococcus aureus.

PURPOSE: This study analyzed rabbit tears for anti-staphylococcal activity, the role of phospholipase A(2) (PLA2) in this reaction, and the ability of enzyme inhibitors to promote bacterial survival. METHODS: Contact lenses with Staphylococcus aureus were applied to scarified rabbit eyes. The colony-forming units (CFU) per cornea or lens were determined and pathology was scored by slit-lamp examination (SLE). The bactericidal activity was measured by incubating bacteria with rabbit tears or PLA2 at 33 degrees or 37 degrees C. Radiolabeled S. aureus was incubated with PLA2 or tears to quantify the release of a membrane component that was identified by thin-layer chromatography. Inhibitors of these reactions were also analyzed. RESULTS: Application of Staphylococcus, on contact lenses, to rabbit corneas resulted in bacterial killing and limited inflammation. Incubation of tears and bacteria (1:1; v/v) in tryptic soy broth at 33 degrees C decreased CFU approximately 4 logs. Tears (> or =30 microl) or PLA2 (> or =30 U) incubated with bacteria in phosphate-buffered saline were bactericidal. PLA2 (> or =0.2 U) or tears (> or =2 microl) cleaved bacterial membranes, liberating arachidonic acid. Spermidine or tetracaine inhibited cleavage of bacterial membranes by tears or PLA2 and spermidine promoted bacterial survival and growth in tears. Tears (60 microl) killed >99% of the bacterial inoculum, whereas bacteria incubated in tears plus spermidine approximately doubled in number. CONCLUSIONS: PLA2 in rabbit tears kills Staphylococcus by hydrolyzing bacterial membranes to release arachidonic acid. Spermidine and tetracaine inhibited PLA2 activity and spermidine protected Staphylococcus from PLA2 in rabbit tears.

Animals↗

Staphylococcus corneal virulence in a new topical model of infection.

PURPOSE: To develop a topical inoculation model of Staphylococcus aureus keratitis in which scarification, contact lenses, and spermidine are used to inhibit the host defenses and to investigate the role of alpha-toxin in this infection. METHODS: An alpha-toxin-positive parent strain (8325-4), its isogenic alpha-toxin-negative mutant (DU1090), and a genetically rescued form of the mutant (DU1090/pDU1212) were bound to rabbit-specific contact lenses, treated with spermidine (50 mM), and applied to scarified rabbit corneas. Eyes were treated topically with spermidine before and after lens application. Eyes were graded for disease by slit lamp examination (SLE) every 6 hours until 24 hours PI (PI), and erosion diameters were measured. Histopathologic changes and colony forming units (CFUs) of bacteria were determined. RESULTS: Spermidine treatment and inoculation of eyes with Staphylococcus on contact lenses resulted in significant increases in both CFUs per cornea (P = 0.0041) and SLE score (P or= 0.1959) multilog increase in CFUs over the inoculum at 24 hours PI. The alpha-toxin-producing strains, 8325-4 and DU1090/pDU1212, caused significantly more disease than the alpha-toxin-deficient mutant DU1090 at 24 hours PI (P <or= 0.0001). Histopathology revealed bacteria in scarified regions of the corneas and, for 8325-4 and DU1090/pDU1212, extensive epithelial sloughing and severe inflammation. CONCLUSIONS: A new topical model of infection has been developed, and alpha-toxin is an important virulence factor in this model.

Animals↗

Effects of 100 mT time varying magnetic fields on the growth of tumors in mice.

The effects of 100-mT, 0.8-Hz square-wave magnetic fields on the growth of chemically induced tumors in mice were investigated. Tumors were initiated using one injection of benzo(a)pyrene (either 0.2 mg or 2.0 mg/animal). Male and female mice (Balb/c, C3H and C57/bl/6 strains) were exposed for 8 h/day from the onset of tumor until death or until the tumor volume reached a predetermined volume. Statistically significant decrease in the rate of tumor growth and increase in survival were observed in all cases. Results are discussed in terms of previous published work and of possible mechanisms.

Animals↗

Lysostaphin treatment of methicillin-resistant Staphylococcus aureus keratitis in the rabbit.

PURPOSE: To determine the efficacy of lysostaphin treatment of methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MRSA) keratitis in a rabbit model. METHODS: The sensitivity to lysostaphin and vancomycin were compared for 34 MRSA and 12 methicillin-sensitive strains. Methicillin-resistant S. aureus strain 301 (MRSA 301) or a methicillin-sensitive strain of low virulence, ISP546, was intrastromally injected into rabbit corneas. Rabbit eyes were treated topically every 30 minutes from 4 to 9 or 10 to 15 hours postinfection with 0.28% lysostaphin or 5.0% vancomycin. Rabbits were killed and corneas were excised and cultured to determine the number of colony forming units (CFU) per cornea. RESULTS: Ninety percent minimal inhibitory concentrations were at least 19-fold lower for lysostaphin than for vancomycin. With early therapy (4 -9 hours postinfection) lysostaphin sterilized all MRSA 301-infected corneas, whereas untreated corneas contained 6.52 log CFU/cornea (P < or = 0.0001). Corneas infected with MRSA 301 and treated similarly with vancomycin retained 2.3 +/-0.85 log CFU/cornea, and none were sterile. When therapy was begun later (10-15 hours postinfection) the residual bacteria in lysostaphin-treated eyes were significantly less numerous than in vancomycin-treated eyes (0.58 +/- 0.34 vs. 5.83 +/- 0.16 log CFU/cornea, respectively; P < or = 0.0001). Three experiments were performed to demonstrate that lysostaphin penetrated the cornea to kill bacteria in vivo; lysostaphin-treated eyes were found to recover from infection, bacteria that did not cause epithelial defects (ISP546) were susceptible to lysostaphin, and inhibition of lysostaphin when harvesting corneas did not alter the observed therapeutic values of lysostaphin. CONCLUSIONS: Lysostaphin is very effective in treating keratitis mediated by methicillin-sensitive or methicillin-resistant S. aureus.

Animals↗

Clarithromycin for experimental Staphylococcus aureus keratitis.

PURPOSE: Clarithromycin, a macrolide antibiotic not previously tested against the common causes of bacterial keratitis, was analyzed for its effectiveness in reducing the number of viable bacteria in a Staphylococcus keratitis model. An in vivo comparison of the effectiveness of clarithromycin to erythromycin, minocycline, and tetracycline for three strains of Staphylococcus aureus was done. METHODS: Rabbit eyes were intrastromally injected with 100 colony forming units of one of three strains of S. aureus. Two strains were methicillin-sensitive (ATCC 25923 and MSSA 309) and one strain methicillin-resistant (COL). Eyes were treated every 30 minutes with 0.3% clarithromycin, erythromycin, tetracycline, or minocycline from 4 to 9 hours postinfection. The number of colony forming units (CFU) per cornea in all eyes was determined at 10 hours postinfection. RESULTS: Vehicle-treated and untreated eyes (controls) contained over 6 logs of CFU per cornea, a value significantly higher than any of the antibiotic-treated eyes (P < or = 0.0001). Clarithromycin or erythromycin therapy significantly decreased the number of CFU per cornea by approximately 5 logs in the eyes infected with the methicillin-sensitive strains and by approximately 4 logs in the eyes infected with the methicillin-resistant strain. Tetracycline and minocycline were also successful in treating these strains, but overall showed less effectiveness than clarithromycin and erythromycin. CONCLUSIONS: Clarithromycin proved to be an effective ocular medication for the therapy of experimental S. aureus keratitis. The effectiveness of clarithromycin in this model and its known effectiveness for a variety of bacterial pathogens suggests a role for this drug as a useful ocular antibiotic.

Animals↗

Serratia marcescens keratitis: strain-specific corneal pathogenesis in rabbits.

PURPOSE: The purpose of this study was to develop an animal model of Serratia keratitis that is suitable to demonstrate the pathology of specific strains. METHODS: Serratia marcescens ocular strains 93-1399-1 and 94-EI-185-2, and an environmental strain (ATCC 14041) were characterized in vitro in terms of their motility, metabolic profiles, ribotypes, and protease production. The strains were then analyzed in the rabbit intrastromal injection model. Slit lamp examination (SLE) and enumeration of bacteria in the cornea was conducted every 6 hours for 30 hours post-infection. In vivo motilities were analyzed by quantification of bacteria in the peripheral and central areas of infected rabbit corneas. RESULTS: All strains were similar in their metabolic activity and production of extracellular proteases. The ocular isolates were distinct from the environmental strain in their ribotyping patterns and in their motility. Each strain grew logarithmically in the cornea up to 6 hours post-infection. SLE scores increased from 0 to 30 hours post-infection for strains ATCC 14041 and 93-1399-1, while the SLE score of strain 94-EI-185-2 reached its maximum at 18 hours post-infection. Strain-specific differences in pathology were noted from 18 to 30 hours post-infection. Strain 94-EI-185-2 produced iritis but only mild corneal changes. Strain 93-1399-1 produced a severe corneal infiltrate encompassing the entire corneal surface as well as severe conjunctival inflammation and iritis. Strain ATCC 14041 produced a localized, severe, exudative corneal abscess that contained infecting bacteria. CONCLUSIONS: A rabbit model of Serratia keratitis was developed in which bacterial growth kinetics and strain-specific ocular pathologic changes were reproducible.

Animals↗

No effect of short-term exposure to GSM-modulated low-power microwaves on benzo(a)pyrene-induced tumours in rat.

PURPOSE: In view of current interest in the biological effects of amplitude-modulated microwaves arising from the rapid development of mobile communications, the effects of low-level microwaves on cancer development were investigated using a rat sarcoma model. MATERIALS AND METHODS: Two-month-old female Sprague-Dawley rats were treated by injection of benzo(a)pyrene and irradiated with GSM (Global System for Mobile)-modulated 900-MHz microwaves in an anechoic chamber at 55 or 200 microW cm(-2) (75 and 270 mW kg(-1) average whole-body SAR, 2h daily for 2 weeks). Rats were exposed from day 20, 40 or 75 after carcinogen injection. Additional groups of rats were sham-exposed in a second anechoic chamber. Anti-phosphatidylinositol autoantibody levels were evaluated in sera to monitor malignant transformation. RESULTS: Microwave exposure had no effect on the development of tumours. No acceleration or delays in tumour onset were observed. Animal survival was not modified and serum autoantibody levels were similar in exposed and sham-exposed groups. CONCLUSION: Low-level GSM microwave exposure of rat bearing benzo(a)pyrene-induced tumours had no effect on auto-antibody levels, tumour appearance and survival. The low exposure levels used here correspond to exposure limits for whole-body exposure of humans.

Animals↗