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Biomedical subjects

H J Linde

Publications and source records attributed to H J Linde.

At least 19 recordsLinked to original sources

In vitro activities of six quinolones and mechanisms of resistance in Staphylococcus aureus and coagulase-negative staphylococci.

Of 94 clinical isolates of Staphylococcus aureus (n = 51) and coagulase-negative staphylococci (CNS) (n = 43), mutations in the quinolone resistance-determining region of topoisomerases GrlA, GrlB, GyrA, and GyrB together with MICs of six quinolones were analyzed. Amino acid substitutions at identical residues (GrlA residues 80 and 84; GyrA residues 84 and 88) were found in S. aureus and CNS. Active efflux, as suggested by blocking by reserpine, contributed substantially to the resistance phenotype in some strains. Among ciprofloxacin, clinafloxacin, levofloxacin, nalidixic acid, trovafloxacin, and sparfloxacin, a 0.5-microg/ml concentration of sparfloxacin discriminated best between strains with two or three mutations and those with no mutations.

4-Quinolones↗

Different subsets of enteric bacteria induce and perpetuate experimental colitis in rats and mice.

Resident bacteria are incriminated in the pathogenesis of experimental colitis and inflammatory bowel diseases. We investigated the relative roles of various enteric bacteria populations in the induction and perpetuation of experimental colitis. HLA-B27 transgenic rats received antibiotics (ciprofloxacin, metronidazole, or vancomycin-imipenem) in drinking water or water alone in either prevention or treatment protocols. Mice were treated similarly with metronidazole or vancomycin-imipenem before or after receiving 5% dextran sodium sulfate (DSS). Germfree transgenic rats were colonized with specific-pathogen-free enteric bacteria grown overnight either in anaerobic or aerobic atmospheres. Nontransgenic rats colonized with anaerobic bacteria served as negative controls. Although preventive metronidazole significantly attenuated colitis in transgenic rats and DSS-treated mice, it had no therapeutic benefit once colitis was established. Ciprofloxacin also partially prevented but did not treat colitis in B27 transgenic rats. In both animal models vancomycin-imipenem most effectively prevented and treated colitis. Germfree transgenic rats reconstituted with enteric bacteria grown under anaerobic conditions had more aggressive colitis than those associated with aerobic bacteria. These results suggest that a subset of resident luminal bacteria induces colitis, but that a complex interaction of commensal aerobic and anaerobic bacteria provides the constant antigenic drive for chronic immune-mediated colonic inflammation.

Animals↗

[Effects of restrictions on use of vancomycin in a German university hospital].

BACKGROUND: Recently, increasing antibiotic resistance has been observed among gram-positive bacteria. However, only few isolates were found to be resistant against glycopeptides. Therefore, internationally accepted guidelines recommend a restricted use of vancomycin and other glycopeptide antibiotics in order to prevent the development of resistance against these clinically important antibiotics. In many countries, the hospital pharmacies play a key role in control and reinforcement of antibiotic formulary restrictions. In Germany, however, the hospital pharmacies usually do not take over such control functions, and most wards keep a stock of regularly used drugs including antibiotics, which makes reinforcement of restrictions difficult. METHODS: In an attempt to achieve a restriction of vancomycin use, the pharmacy of our university hospital was advised to deliver vancomycin to the wards only on request with a special order form signed by an attending, individually for every patient who should receive vancomycin. The efficacy of this restriction measure was evaluated in 3-month periods before and after the restriction became effective. RESULTS: Hospitalwide, this led to a 20.1% reduction of i.v. vancomycin and an 85.7% reduction of oral vancomycin use per 1000 patient days. If the hematology/oncology units were not considered, the reduction of i.v. vancomycin use was 41.8%, and the total use after the restriction 24.2 g per 1000 patient days. Microbiology results which justified the use of vancomycin decreased by 8.3% (10.9% hematology/oncology units not considered) between the 2 observation periods. Assuming a 7-day mean course of i.v. vancomycin therapy, the empirical use of i.v. vancomycin decreased from 39.9% to 8% after the restriction had been instituted. CONCLUSION: Allowing only experienced physicians (attendings) to decide on the use of vancomycin therapy, proved in our experience to be an effective measure to reduce unnecessary vancomycin use.

Drug Utilization↗

Improved detection of microorganisms by polymerase chain reaction in delayed endophthalmitis after cataract surgery.

OBJECTIVE: To evaluate whether the use of polymerase chain reaction (PCR) improves the identification of the causative pathogen in eyes developing delayed endophthalmitis after cataract surgery. DESIGN: Prospective, noncomparative case series. PARTICIPANTS: Consecutive series of 25 eyes with the clinical diagnosis of delayed endophthalmitis after cataract. MAIN OUTCOME MEASURE: Presence of bacterial or fungal DNA in aqueous humor and vitreous samples. RESULTS: In the aqueous humor the causative pathogen was identified in 84% (n = 21) of the eyes by PCR compared with 0% by diagnostic culture and 0% by microscopy. In the vitreous samples the pathogen was identified in 92% (n = 23) of the eyes by PCR compared with 24% by diagnostic culture (n = 6) and 0% by microscopy. CONCLUSIONS: PCR is useful for the identification of the causative pathogen in delayed endophthalmitis and had a higher rate of positive identification of the causative organism than microscopy or diagnostic culture.

Aqueous Humor↗

[Disciform keratatis caused by Bartonella henselae infection: detection of a rare ocular complication of cat-scratch disease with PCR].

BACKGROUND: Cat scratch disease (CSD) is a common infectious disease, however its association with a disciforme keratitis is to our knowledge a previously unreported ocular complication. PATIENT: A 65-year old female was treated unsuccessfully with topical antibiotics, acyclovir and corticosteroids for several weeks because of a disciforme keratitis. Repeatedly, microbiological investigations (microscopy and culture) were negative. 16S rDNA-PCR with subsequent DNA-sequencing on material obtained by corneal scrape identified Bartonella henselae. Based on this result we have initiated a treatment with topical cephotaxim and within a short period of time the cornea healed with a paracentral scar. CONCLUSION: 16S rDNA-PCR was very helpful in identifying the intracellular sited bacterium Bartonella henselae as the causative pathogen of an unusual form of disciforme keratitis.

Aged↗

A bacteria-induced switch of sympathetic effector mechanisms augments local inhibition of TNF-alpha and IL-6 secretion in the spleen.

It is believed that an inflammation-induced activation of the CNS leads to an inhibition of overshooting immune responses to prevent extensive local cytokine secretion. However, immunosuppression by the sympathetic nervous system may be unfavorable when bacteria are present locally and when TNF-alpha is necessary to overcome infection. We now report in a superfusion model, using mouse spleen slices, that although local Pseudomonas aeruginosa increased splenic TNF-alpha and IL-6 secretion severalfold over basal levels, electrically released neurotransmitters attenuated cytokine secretion to similar basal level as under bacteria-free conditions. Bacteria reversed noradrenergic inhibitory effector mechanisms: Under bacteria-free conditions, TNF-alpha secretion was very low and IL-6 secretion was mainly inhibited by alpha2-adrenoreceptor ligation. In the presence of bacteria, TNF-alpha and IL-6 secretion were high and IL-6 secretion was mainly inhibited by beta-adrenoreceptor ligation. The alpha- to beta-adrenoswitch of IL-6 inhibition in the presence of bacteria was mediated by the prior adrenergic regulation of TNF-alpha. In vivo, chemical abrogation of sympathetic inhibition reduced accumulation of bacteria in the spleen, which depended at least in part on TNF-alpha. This suggests that activation of the sympathetic nervous system may be a forerunner for accumulation of bacteria in tissue and consecutively sepsis due to intensified inhibition of TNF-alpha secretion.

Animals↗

In vivo increase in resistance to ciprofloxacin in Escherichia coli associated with deletion of the C-terminal part of MarR.

We recovered two isolates (EP1 and EP2) of Escherichia coli from the same patient that had identical pulsed-field gel electrophoresis patterns but required different MICs of ciprofloxacin (CIP): 16 and 256 mg/liter for EP1 and EP2, respectively. Both isolates had mutations in the quinolone resistance-determining regions of GyrA (Ser83Leu and Asp87Tyr) and ParC (Ser80Ile), but not in those regions of GyrB or ParE. Isolate EP2 was also more resistant to chloramphenicol, tetracyclines, cefuroxime, and organic solvents. A deletion of adenine (A) 1821 was found in marR of isolate EP2, which resulted in an 18-amino-acid C-terminal deletion in the MarR protein. The causative relationship between DeltaA1821 and the Mar phenotype was demonstrated both by the replacement of the wild-type marR by marR DeltaA1821 in isolate EP1 and by complementation with the wild-type marR in trans in isolate EP2. In isolate EP2 complemented with wild-type marR, susceptibility to chloramphenicol was restored completely, whereas susceptibility to CIP was restored only incompletely. Northern blotting demonstrated increased expression of marA and acrAB but not of soxS in isolate EP2 compared to EP1. In conclusion, the deletion of A1821 in marR in the clinical isolate EP2 caused an increase in the MICs of CIP and unrelated antibiotics. Presumably, the C-terminal part of MarR is necessary for proper repressor function.

Anti-Infective Agents↗

Rapid identification of methicillin-resistant Staphylococcus aureus and simultaneous species confirmation using real-time fluorescence PCR.

A duplex LightCycler PCR assay targeting the mecA gene and a Staphylococcus aureus-specific marker was used to test 165 S. aureus strains and 80 strains of other bacterial species. Within an assay time of 60 min plus 10 min for sample preparation, S. aureus as well as the presence or absence of the mecA gene was correctly identified.

Bacterial Proteins↗

Varying cecal bacterial loads influences colitis and gastritis in HLA-B27 transgenic rats.

BACKGROUND & AIMS: Recent data support an important role of resident luminal bacteria in experimental colitis. We determined how altered cecal bacterial loads influence colitis and gastritis. METHODS: A cecal self-filling blind loop (SFBL) was created or the cecum was excluded from the fecal stream in specific pathogen-free HLA-B27 transgenic (TG) rats with early colitis and in nontransgenic (nonTG) littermates; controls underwent sham operation (SHAM). Luminal bacterial concentrations were determined by culture and counting chamber. RESULTS: TG rats with SFBL had more severe cecal inflammation and leukocytosis than TG SHAM controls. TG excluded rats with low cecal bacterial loads had no cecal inflammation and less colitis and gastritis than SHAM controls, despite having normal distal colonic and gastric bacterial concentrations. Metronidazole attenuated cecal inflammation and eliminated Bacteroides in SFBL TG rats. NonTG SFBL rats had mild cecal inflammation and no gastritis and colitis. The ratio of total anaerobic to aerobic bacteria was 1000-fold greater in SFBL than in SHAM rats, with a 10,000-fold increased ratio of Bacteroides spp. to aerobes. CONCLUSIONS: The luminal bacterial load and composition determines the activity of cecal inflammation in genetically susceptible hosts. Lowering cecal bacterial concentrations can diminish inflammation in remote organs.

Animals↗

Identification of Yersinia species by the Vitek GNI card.

The Vitek GNI card was used to identify 212 isolates of 10 Yersinia species. Identification was correct for 96.3% of the isolates (156 of 162) to the genus level and for 57.4% of the isolates (93 of 162) to the species level for Yersinia spp. listed in the Vitek database. We recommend additional identification methods for isolates assigned to the genus Yersinia by the Vitek system.

Bacteriological Techniques↗

Diagnosis of infectious endophthalmitis after cataract surgery by polymerase chain reaction.

PURPOSE: To ascertain whether the use of the polymerase chain reaction (PCR) technique leads to more rapid diagnosis of infectious endophthalmitis after cataract surgery. SETTING: University Eye Clinic Regensburg, Germany. METHODS: The aqueous humor and vitreous of 16 eyes with infectious endophthalmitis (10 acute, 6 delayed) were evaluated by microscopy, diagnostic culture, and PCR to detect the infectious agent. RESULTS: Microscopy of the vitreous was positive in 3 eyes and the culture media results were positive in 7 eyes, all with acute endophthalmitis. Significantly fewer positive results were obtained in the aqueous humor. Using PCR, an infectious agent was detected in the aqueous humor of all 16 eyes and in the vitreous of 14. The vitreous sample was negative in 2 eyes with delayed endophthalmitis. CONCLUSIONS: Detection of the infectious agent was more successful using PCR than using conventional microbiological tests, especially in the diagnosis of delayed endophthalmitis where the pathogen was detected in the aqueous humor in all eyes.

Acute Disease↗

[Listeria monocytogenes-induced endogenous endophthalmitis in an otherwise healthy patient: PCR-assisted rapid diagnosis as the basis for successful therapy].

BACKGROUND: Listeria monocytogenes is a rare cause of endogenous endophthalmitis. Only 14 cases are published in the literature so far. All eyes showed similar clinical features and profound visual loss. PATIENTS AND METHODS: We report on a case of an otherwise healthy 73-year-old male. He was referred to our hospital because of acute hypopyoniritis with secondary glaucoma. Within a few hours the severity of the intraocular infection increased dramatically resulting in the clinical picture of an acute endophthalmitis. RESULTS AND CONCLUSION: Early identification of the causative pathogen in the aqueous humor after anterior chamber paracenthesis using polymerase chain reaction (PCR) and the initiation of a specific, systemic antibiotic medication resulted in a complete recovery of visual acuity.

Acute Disease↗

Development and characterization of a murine monoclonal antibody reactive with a 64 kDa somatic antigen of Burkholderia cepacia.

Monoclonal antibodies (MAbs) to Burkholderia cepacia were produced from mice immunized with inactivated whole-cell antigen. For screening of resulting MAbs an enzyme-linked immunosorbent assay (ELISA) was used. A stable hybridoma cell line (BC-2) producing specific antibodies to a 64 kDa somatic antigen from B. cepacia was established. In ELISA and immunoblotting analysis the MAb BC-2 recognized all tested strains of B. cepacia whereas no cross-reaction with 32 Pseudomonas aeruginosa strains was found. From a wide range of other bacteria only strains of the species Burkholderia mallei, Burkholderia pseudomallei, and Burkholderia gladioli showed cross-reactions. The MAb BC-2 will be used to develop a diagnostic assay for the identification of B. cepacia and B. gladioli, important agents of nosocomial infections in immunocompromised patients suffering especially from cystic fibrosis (CF).

Animals↗

[Rapid diagnosis of infectious endophthalmitis using polymerase chain reaction (PCR): a supplement to conventional microbiological diagnostic methods].

BACKGROUND: Endophthalmitis is, although relatively rare, a serious intraocular infection, which could result in a loss of visual function. Therefore, the rapid diagnosis and initiation of the appropriate treatment is of critical importance. To date, approximately 60 percent of eyes with a clinical diagnosis of infectious endophthalmitis show a positive microscopic or culture result. By using the very sensitive polymerase chain reaction this number might increase. PATIENTS AND METHODS: In a series of 12 eyes with infectious endophthalmitis we have performed microscopic investigations, diagnostic culture and polymerase chain reaction in aqueous humor and vitreous in order to detect the infectious agent. RESULT: Microscopic investigations showed a positive result in the vitreous of 3 eyes. This number improved to 6 eyes using culture media. Significant less positive results were obtained in the aqueous humor. The infectious agent could be detected in the aqueous humor in all 12 eyes and in the vitreous in 9 eyes by PCR. Only in 2 eyes with a delayed endophthalmitis the vitreous was negative. CONCLUSIONS: The detection of the infectious agents was more successful using PCR compared to conventional microbiological tests. In particular, for the diagnosis of delayed endophthalmitis PCR proves to be very superior. In all cases of delayed endophthalmitis the pathogen could be detected in the aqueous humor.

Aqueous Humor↗

[Eradication of Legionella from warm water systems--documentation of our own experiences with thermal disinfection].

A nosocomial case of Legionellosis in a recently built hospital was the reason for an investigation and thermal disinfection of the complete warm water distribution system. Furthermore weak points in the tubing of the warm water system, which promoted the contamination of potable water, were eliminated as far as technically possible. A lasting reduction of the numbers of Legionella spp. isolated could be measured (factor 10-1000), but a complete decontamination of the warm water distribution system was not possible. As Legionella spp. may cause serious infections in immunodeficient patients, additional measures for the disinfection of the hospital water distribution system must be taken into consideration.

Cross Infection↗

Uptake and killing of Candida by human peritoneal macrophages and amphotericin B.

The effects of amphotericin B at subinhibitory and inhibitory concentrations on ingestion and intracellular killing of C. albicans ATCC 10,231 and C. tropicalis ATCC 13,803 by human peritoneal macrophages in vitro was investigated. Peritoneal macrophages were harvested from overnight peritoneal dialysate of 26 patients undergoing regular continuous ambulatory peritoneal dialysis (CAPD) using a new simple isolation technique. Macrophages were suspended with Candida (1:2-3) together with pooled human serum and with or without amphotericin B at various concentrations. Vital staining with acridine orange at a very low concentration using the metachromatic property of the dye allowed simultaneous assessment of ingestion and intracellular viability of the yeasts. Counts of Candida in 100 macrophages were performed at 1, 2, 3, 4, 6 and 24 h under a fluorescence microscope at 1000x and the ratios of living to dead intracellular Candida were calculated. Amphotericin B was added at concentrations of 0.1, 1 and 10 times the MIC. Ingestion was rapid and complete, while intracellular killing ranged from 4-69% for C. albicans and from 9-48% for C. tropicalis. Amphotericin B at 1 MIC enhanced the killing of C. tropicalis (factor 1.32) but reduced killing of C. albicans (factor 0.6) after 6 h.

Adult↗