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Biomedical subjects

J Bille

Publications and source records attributed to J Bille.

At least 73 records · Page 4Linked to original sources

Typing Listeria monocytogenes isolates from fish products and human listeriosis cases.

Seventy-two Listeria monocytogenes isolates originating from 10 different fish products of 12 producers and 47 isolates from human listeriosis cases were typed by serotyping and multilocus enzyme electrophoresis. Seventy-five of these isolates were further subtyped by restriction analysis of genomic DNA with the enzyme XhoI and by pulsed-field gel electrophoresis using the enzymes ApaI and SmaI. The results show that several L. monocytogenes clones identified by multilocus enzyme electrophoresis are frequently found in fish products of different origins. One of these clones is the same as another previously shown to be frequently associated with meat and meat products. The epidemic-associated electrophoretic type 1 was only rarely found in fish products. No association was found between any type of fish product and a particular lineage of L. monocytogenes. Both long-term persistence of a strain and simultaneous presence of several clearly distinct strains in the products of single producers were observed. The comparison of L. monocytogenes isolates from human clinical listeriosis cases in Switzerland and those from imported fish products by use of multilocus enzyme electrophoresis showed that they do not form two clearly distinct lineages but nevertheless belong to two separate populations. None of the 48 subtypes distinguished by the combination of all four typing methods could be found in both populations of human origin and those of fish origin.

Animals↗

Typing of Pneumocystis carinii f. sp. hominis by single-strand conformation polymorphism of four genomic regions.

To better investigate Pneumocystis carinii f. sp. hominis epidemiology, we have developed a molecular typing method. Because of the limited genetic variability of the P. carinii hominis genome, a multitarget approach was used. Four variable regions of the genome were amplified by PCR, polymorphism in each region was assessed by the single-strand conformation polymorphism (SSCP) technique, and the results for the four regions of each patient were combined. Bronchoalveolar lavage specimens collected from 11 patients were examined. Four patients were probably infected by a single strain, since their specimens yielded simple SSCP patterns (two bands corresponding to one allele). The combinations of these patterns were unique, suggesting that the strains which infected these patients were different. For the other seven patients, complex patterns were found (three or four bands corresponding to two alleles). The presence of more than one allele of a region in a patient is likely to be due to coinfection. Polymorphism was also assessed by sequencing, which revealed variations at nucleotide positions previously reported to vary. About half of the observed alleles had already been reported by laboratories in different countries. Multitarget typing of P. carinii hominis by PCR-SSCP should allow investigation of strain diversity and thus be useful for future epidemiological studies.

AIDS-Related Opportunistic Infections↗

Widespread geographic distribution of oral Candida dubliniensis strains in human immunodeficiency virus-infected individuals.

Candida dubliniensis is a recently identified chlamydospore-positive yeast species associated with oral candidiasis in human immunodeficiency virus (HIV)-infected (HIV+) patients and is closely related to Candida albicans. Several recent reports have described atypical oral Candida isolates with phenotypic and genetic properties similar to those of C. dubliniensis. In this study 10 atypical chlamydospore-positive oral isolates from HIV+ patients in Switzerland, the United Kingdom, and Argentina and 1 isolate from an HIV-negative Irish subject were compared to reference strains of C. albicans and Candida stellatoidea and reference strains of C. dubliniensis recovered from Irish and Australian HIV+ individuals. All 11 isolates were phenotypically and genetically similar to and phylogenetically identical to C. dubliniensis. These findings demonstrate that the geographical distribution of C. dubliniensis is widespread, and it is likely that it is a significant constituent of the normal oral flora with the potential to cause oral candidiasis, particularly in immunocompromised patients.

AIDS-Related Opportunistic Infections↗

In vitro activity of cefpirome against microorganisms isolated in haematology, oncology and intensive care units in Switzerland.

The in vitro activity of cefpirome, a new parenteral fourth-generation cephalosporin, was investigated in the 5 university hospitals of Switzerland, and compared to 9 other antibiotics mainly used in hospitals, such as ceftazidime, ceftriaxone, cefotaxime, piperacillin, imipenem, gentamicin, vancomycin, ciprofloxacin and ofloxacin. A total number of 992 strains collected only from intensive care units and haematology-oncology units were tested by microdilution according to NCCLS. Cefpirome showed an excellent activity against all Enterobacteriaceae (MIC90 = 4 mg/l), methicillin-susceptible staphylococci (MIC90 = 1 mg/l), Streptococcus pneumoniae (MIC90 = 0.25 mg/l) and Haemophilus influenzae (MIC90 = 0.12 mg/l) isolates. Its activity was superior to that of third-generation cephalosporins against cephalosporinase-depressed mutants of Enterobacter cloacae and Citrobacter freundii isolates (MIC90 > 32 mg/l for third-generation cephalosporins vs 4 mg/l for cefpirome). The MICs of cefpirome of 3 strains of Klebsiella spp. with an extended-spectrum-beta-lactamase were lower (MIC90 = 2 mg/l) than those of third-generation cephalosporins (MICs90 > 32 mg/l). Against Pseudomonas aeruginosa cefpirome was as active as ceftazidime. The activity of cefpirome was poor against methicillin-resistant staphylococci, enterococci and nosocomial Gram-negative bacteria such as Stenotrophomonas maltophilia.

Anti-Bacterial Agents↗

Foodborne listeriosis.

Various epidemiological investigations of outbreaks and sporadic cases have clearly demonstrated that the consumption of contaminated food is responsible for a high proportion of listeriosis cases and Listeria monocytogenes has been increasingly recognized as an important foodborne pathogen over the last 15 years. The emergence of listeriosis is the result of complex interactions of different factors: medical progress which increases the lifespan and allows immunodeficient people to survive, expansion of the food industry and cold storage systems as well as changes in food habits. None of these factors on its own is entirely responsible. Considerable research has attempted to characterize the organism, define the magnitude of the public health problem and its impact on the food industry, identify the risk factors associated with the disease, and devise appropriate control strategies. Nevertheless, a number of crucial questions remains incompletely elucidated (extent of the foodborne transmission of listeriosis, health status of apparently "healthy patients" with the possible role of an intercurrent infection or genetic susceptibility, how to distinguish highly virulent from less virulent strains of L. monocytogenes, factors contributing to the emergence of outbreaks, the possible role of healthy carriers in the epidemiology of listeriosis, etc.). To investigate the complexity of listeriosis requires the close collaboration of clinicians, epidemiologists, clinical and food microbiologists, food scientists and the food industry. A large amount of data has been accumulated during the past 10 years but more research is required to elucidate the epidemiology of the disease and the virulence of the causative agent.

Disease Outbreaks↗

Immunocytochemical reaction of a haemocyanin antibody in the midgut gland of Nautilus (Cephalopoda, Tetrabranchiata).

The branchial gland of the dibranchiate cephalopods is described as the site of haemocyanin synthesis. Because there is no equivalent to this organ in tetrabranchiate cephalopods the localization of haemocyanin synthesis remained unknown for a long time. In this study we could confirm the conclusions from preliminary investigations concerning the copper content of the midgut gland of Nautilus, which gave the first indications for a possible localization of haemocyanin synthesis in this organ. We developed a polyclonal antibody against Nautilus haemocyanin, tested its specificity, and used it on ultra-thin sections of the tissue of the midgut gland. It could be shown that there is a clear immunogold precipitation only on the triangular basal cells in the terminal alveoli. All the other types of cell in this organ were free of any immunoreactivity. It can be supposed that the triangular basal cells in the terminal alveoli of the midgut gland are the sites of haemocyanin synthesis in Nautilus.

Animals↗

Ablation of neural tissue by short-pulsed lasers--a technical report.

The basis for most laser applications in neurosurgery is the conversion of laser light into heat when the incident laser beam is absorbed by the tissue. Irradiation of neural tissue with laser light therefore leads to its thermal damage. However, due to the diffusion of heat energy into the surrounding tissue, often there is thermal damage to neural tissue outside the area of the target volume. These are the characteristics of thermal laser/tissue interaction. In this paper we discuss how we used three different short-pulsed lasers to achieve non-thermal ablation of neural tissue. Three different short-pulsed lasers were used to generate ultrashort laser pulses in the picosecond to femtosecond range. The interaction of such laser pulses with tissue was predicted to be nonthermal. The short-pulsed lasers were used for the ablation of neural tissue using an in vitro calf brain model. The histopathological examination of the lesions revealed that the neural tissue had been removed very precisely without any sign of thermal damage to the surrounding tissue.

Animals↗

Typing of Listeria monocytogenes by monocin and phage receptors.

One hundred strains of Listeria monocytogenes from both sporadic and epidemic cases were typed by monocin production combined with phage receptor and reverse phage receptor methods. The monocin-phage combination gave 72 types with 100% typability and 97% reproducibility. The results were compared to those of serotyping, phage typing, ribotyping, multilocus enzyme electrophoresis, restriction enzyme analysis and RAPD (random amplification of polymorphic DNA). The monocin/phage types were comparable in terms of discrimination with other methods for epidemiological investigations. The index of discrimination of using the monocin typing and phage receptor/reverse phage receptor method combination (0.99) for both the 87 epidemiologically unrelated strains and the epidemiologically important serogroup 4 strains was the highest of the seven different methods analysed. This combination of methods was simple, highly discriminatory and reproducible and can be carried out in a non-specialized laboratory. However, like most of the other Listeria typing methods, both the method and the indicator test strains need to be standardized.

Bacterial Typing Techniques↗

Quantitative antibiogram as a typing method for the prospective epidemiological surveillance and control of MRSA: comparison with molecular typing.

OBJECTIVE: Evaluation of the quantitative antibiogram as an epidemiological tool for the prospective typing of methicillin-resistant Staphylococcus aureus (MRSA), and comparison with ribotyping. METHODS: The method is based on the multivariate analysis of inhibition zone diameters of antibiotics in disk diffusion tests. Five antibiotics were used (erythromycin, clindamycin, cotrimoxazole, gentamicin, and ciprofloxacin). Ribotyping was performed using seven restriction enzymes (EcoRV, HindIII, KpnI, PstI, EcoRI, SfuI, and BamHI). SETTING: 1,000-bed tertiary university medical center. RESULTS: During a 1-year period, 31 patients were found to be infected or colonized with MRSA. Cluster analysis of antibiogram data showed nine distinct antibiotypes. Four antibiotypes were isolated from multiple patients (2, 4, 7, and 13, respectively). Five additional antibiotypes were isolated from the remaining five patients. When analyzed with respect to the epidemiological data, the method was found to be equivalent to ribotyping. Among 206 staff members who were screened, six were carriers of MRSA. Both typing methods identified concordant of MRSA types in staff members and in the patients under their care. CONCLUSIONS: The quantitative antibiogram was found to be equivalent to ribotyping as an epidemiological tool for typing of MRSA in our setting. Thus, this simple, rapid, and readily available method appears to be suitable for the prospective surveillance and control of MRSA for hospitals that do not have molecular typing facilities and in which MRSA isolates are not uniformly resistant or susceptible to the antibiotics tested.

Bacterial Typing Techniques↗

Susceptibilities of Candida albicans multidrug transporter mutants to various antifungal agents and other metabolic inhibitors.

Some Candida albicans isolates from AIDS patients with oropharyngeal candidiasis are becoming resistant to the azole antifungal agent fluconazole after prolonged treatment with this compound. Most of the C. albicans isolates resistant to fluconazole fail to accumulate this antifungal agent, and this has been considered a cause of resistance. This phenomenon was shown to be linked to an increase in the amounts of mRNA of a C. albicans ABC (ATP-binding cassette) transporter gene called CDR1 and of a gene conferring benomyl resistance (BENr), the product of which belongs to the class of major facilitator multidrug efflux transporters (D. Sanglard, K. Kuchler, F. Ischer, J. L. Pagani, M. Monod, and J. Bille, Antimicrob. Agents Chemother. 39:2378-2386, 1995). To analyze the roles of these multidrug transporters in the efflux of azole antifungal agents, we constructed C. albicans mutants with single and double deletion mutations of the corresponding genes. The mutants were tested for their susceptibilities to these antifungal agents. Our results indicated that the delta cdr1 C. albicans mutant was hypersusceptible to the azole derivatives fluconazole, itraconazole, and ketoconazole, thus showing that the ABC transporter Cdr1 can use these compounds as substrates. The delta cdr1 mutant was also hypersusceptible to other antifungal agents (terbinafine and amorolfine) and to different metabolic inhibitors (cycloheximide, brefeldin A, and fluphenazine). The same mutant was slightly more susceptible than the wild type to nocodazole, cerulenin, and crystal violet but not to amphotericin B, nikkomycin Z, flucytosine, or pradimicin. In contrast, the delta ben mutant was rendered more susceptible only to the mutagen 4-nitroquinoline-N-oxide. However, this mutation increased the susceptibilities of the cells to cycloheximide and cerulenin when the mutation was constructed in a delta cdr1 background. The assay used in the present study could be implemented with new antifungal agents and is a powerful tool for assigning these substances as putative substrates of multidrug transporters.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Molecular epidemiology of fluoroquinolone-resistant Escherichia coli bloodstream isolates from patients admitted to European cancer centers.

Previous reports have suggested an increasing incidence of highly fluoroquinolone-resistant Escherichia coli causing bacteremia among cancer patients on prophylactic therapy. We used genotyping by pulsed-field gel electrophoresis of chromosomal DNA digests and random amplified polymorphic DNA fingerprinting to study clonal relationships among such isolates obtained at 10 cancer centers located across Europe and the Middle East. Analysis by both methods indicated that isolates from different centers were genotypically unrelated to each other. There were five centers from which more than one individual patient isolate was available, and most demonstrated significant within-center genetic diversity of strains. Strains shared among patients could be identified at two centers. At the center with the largest number of bloodstream isolates from cancer patients available, fluoroquinolone-resistant control isolates from surgical patients and fluoroquinolone-susceptible control isolates from patients admitted to medical services during the same time period were unrelated to resistant cancer patient isolates and to each other as well. A substantial number of fluoroquinolone-resistant isolates (19 of 58) were nontypeable by pulsed-field gel electrophoresis. Fluoroquinolone resistance was commonly associated with multiple antibiotic resistance to chemically unrelated antibacterial agents irrespective of the origin of the isolates.

Anti-Infective Agents↗

Typing Candida albicans oral isolates from human immunodeficiency virus-infected patients by multilocus enzyme electrophoresis and DNA fingerprinting.

A total of 189 Candida albicans isolates have been typed by multilocus enzyme electrophoresis. The results obtained confirm the clonal mode of reproduction of C. albicans. The C. albicans populations found in the oropharynx of human immunodeficiency virus (HIV)-infected patients, in the oropharynx of healthy carriers, or in association with invasive candidiasis could not be distinguished. No clone or group of clones could be associated with the appearance of clinical disorders or with a reduced in vitro susceptibility to the antifungal agent fluconazole. Multiple and sequential oral isolates from 24 HIV-infected patients were also typed by restriction enzyme analysis with the enzymes EcoRI and HinfI and by use of the Ca3 repetitive probe. The results obtained by the combination of all three typing methods show that all but one patient each carried a unique major C. albicans clone in their oropharynx. The 21 patients with sequential isolates had the same C. albicans clones in their throats during recurrent oropharyngeal candidiasis episodes, independently of clinical status or of changes of in vitro susceptibility to fluconazole. Finally, several isolates of the same C. albicans clone found simultaneously in the oropharynx of a patient may present different levels of susceptibility to fluconazole.

AIDS-Related Opportunistic Infections↗

Subtyping Listeria monocytogenes isolates genetically related to the Swiss epidemic clone.

Macrorestriction analysis by pulsed-field gel electrophoresis was used to assess the diversity of strains within the epidemic-associated electrophoretic type 1 (ET1) clone of Listeria monocytogenes. For this purpose, a total of 144 isolates from Switzerland shown by multilocus enzyme electrophoresis to belong to the ET1 were examined. These isolates were subtyped by macrorestriction analysis using the enzymes ApaI and SmaI and field inversion gel electrophoresis. Among these 144 isolates, 45 were isolated in human listeriosis cases of the postepidemic period of 1988 to 1993 and 44 were isolated in animal listeriosis cases of the same period. Forty-seven isolates were from the epidemic period of 1983 to 1987, and eight additional isolates were from cattle from two different farms. Twenty-nine different subtypes could be identified among the 144 isolates tested. Five major subtypes were found more frequently than the others during the postepidemic period, both in humans and in animals. Two of these subtypes had been previously implicated in outbreaks of listeriosis, thus suggesting that particular pulsed-field gel electrophoresis subtypes may be frequently associated with disease in humans and animals. Two of these frequent subtypes were also suspected to be related to small clusters of listeriosis cases during the postepidemic period. The results obtained by typing epidemiologically related isolates from different animals within the same farms and from different body sites of a given patient confirmed the potential of macrorestriction analysis for epidemiological studies restricted to short periods of time and to small number of isolates. The analysis of 47 isolates related to the Swiss listeriosis epidemic period of 1983 to 1987 and the use of Southern blotting and hybridization experiments show that the interpretation of relatedness between isolates presenting slightly different macrorestriction patterns may be more complex than commonly accepted. In such cases, careful interpretation of the potential molecular mechanisms leading to the differences observed between patterns is necessary.

Animals↗

Bacterial overgrowth during treatment with omeprazole compared with cimetidine: a prospective randomised double blind study.

BACKGROUND: Gastric and duodenal bacterial overgrowth frequently occurs in conditions where diminished acid secretion is present. Omeprazole inhibits acid secretion more effectively than cimetidine and might therefore more frequently cause bacterial overgrowth. AIM: This controlled prospective study compared the incidence of gastric and duodenal bacterial overgrowth in patients treated with omeprazole or cimetidine. METHODS: 47 outpatients with peptic disease were randomly assigned to a four week treatment regimen with omeprazole 20 mg or cimetidine 800 mg daily. Gastric and duodenal juice were obtained during upper gastrointestinal endoscopy and plated for anaerobic and aerobic organisms. RESULTS: Bacterial overgrowth (> or = 10(5) cfu/ml) was present in 53% of the patients receiving omeprazole and in 17% receiving cimetidine (p < 0.05). The mean (SEM) number of gastric and duodenal bacterial counts was 6.0 (0.2) and 5.0 (0.2) respectively in the omeprazole group and 4.0 (0.2) and 4.0 (0.1) in the cimetidine group (p < 0.001 and < 0.01; respectively). Faecal type bacteria were found in 30% of the patients with bacterial overgrowth. Basal gastric pH was higher in patients treated with omeprazole compared with cimetidine (4.2 (0.5) versus 2.0 (0.2); p < 0.001) and in patients with bacterial overgrowth compared with those without bacterial overgrowth (5.1 (0.6) versus 2.0 (0.1); p < 0.0001). The nitrate, nitrite, and nitrosamine values in gastric juice did not increase after treatment with either cimetidine or omeprazole. Serum concentrations of vitamin B12, beta carotene, and albumin were similar before and after treatment with both drugs. CONCLUSIONS: These results show that the incidence of gastric and duodenal bacterial overgrowth is considerably higher in patients treated with omeprazole compared with cimetidine. This can be explained by more pronounced inhibition of gastric acid secretion. No patient developed signs of malabsorption or an increase of N-nitroso compounds. The clinical significance of these findings needs to be assessed in studies with long-term treatment with omeprazole, in particular in patients belonging to high risk groups such as HIV infected and intensive care units patients.

Adult↗

[Systemic mycotic infections: epidemiology and diagnostic criteria].

Among the systemic mycotic infections, disseminated candidiasis is the most frequent and is constantly on the increase. Candidiasis occurs in patients with particular risks, i.e., in oncologic neutropenic patients and in surgical patients after intraabdominal surgery. Independent predictive factors are multiple prior antibiotic therapy, the presence of intravascular devices and previous colonization by Candida, as well as the rupture of skin or cutaneous anatomical barriers. C. albicans is still the predominant species. Diagnosis of disseminated candidiasis is difficult and sometimes delayed. It relies essentially on a high index of suspicion, on blood cultures and the analysis of other samples (biopsies) by a combination of microbiological and anatomopathological procedures. The detection of Candida antigens, antibodies or metabolites still suffers from suboptimal sensitivity and specificity. Molecular diagnosis by amplification (PCR) is promising. The reservoir of Candida is mainly the human digestive tract, and a colonization stage usually precedes an established infection. Transmission of Candida by human carriage or through the environment could play an important role, particularly in intensive care units.

Antigens, Fungal↗

[Pneumococcal bacteremia: what is new?].

85 patients who displayed Streptococcus pneumoniae (S. pneumoniae) bacteremia during hospitalization at the CHUV between January 1990 and December 1991 are reviewed retrospectively to reassess the importance of this pathology after the introduction in Switzerland of antipneumococcal vaccination. The data were compared with those obtained from a similar study at Lausanne between 1974 and 1978. Epidemiology, underlying diseases (present in 82% of patients), clinical findings (78% pneumonia, 8% meningitis, 14% bacteremia without detected primary focus) and mortality (31%) were comparable in the two series. There were, however, points of difference. First, the incidence of pneumococcal bacteremia increased between 20 and 40 years, affecting HIV positive patients in particular, with high mortality. Second, nosocomial pneumococcal bacteremia was relatively frequent (7%) and particularly severe, with very high mortality. Third, 17% of strains proved resistant to one or more antibiotics. Finally, use of the antipneumococcal vaccine is not widespread since only one of the 85 patients had been vaccinated whereas 82% presented a recognized indication for the vaccine. In conclusion, pneumococcal bacteremia remains frequent, involves major mortality and is more often due to resistant strains. These are important arguments in favour of vaccinating patients at risk.

AIDS-Related Opportunistic Infections↗