- Volume 43, Issue 5, 1995
Volume 43, Issue 5, 1995
- Review Article
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Staphylococcal scalded skin syndrome
More LessSummaryStaphylococcal scalded skin syndrome (SSSS) is a recognised clinical entity that affects primarily the very young and, in rare cases, the very old or the immunocompromised. Koch’s postulates have been fulfilled in that: (i) Staphylococcus aureus is isolated from every case; (ii) S. aureus can reproduce the syndrome in an experimental animal model; (iii) a specific extracellular toxin can reproduce the syndrome; and (iv) antibody to the toxin can protect experimental animals. Although exfoliative toxin (ET) is responsible for the skin loosening seen in SSSS, it does not account for all the symptoms of the disease. Purified ET does not cause erythema in either neonatal mice or man, and the lesions are not painful unless the loosened epidermis is removed. This suggests that other factors, e.g., δ-haemoiysin, are involved in the pathogenesis of this condition. Although much has been learned about the pathogenesis of the syndrome, we are still largely ignorant of the factors which govern host resistance to SSSS (i.e., intoxication by ET-producing strains of S. aureus). It is fortunate from the patient’s point of view that the aetiological agent can be destroyed readily by the use of appropriate antibiotic therapy.
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- Antimicrobial Agents
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Efficacy of sustained release ciprofloxacin microspheres against device-associated Pseudomonas aeruginosa biofilm infection in a rabbit peritoneal model
More LessSummaryThe relative effectiveness of a poly(L-lactic acid) ciprofloxacin hydrochloride (CIP) microsphere formulation (250–425 μm) against peritoneal implanted biofilm of Pseudomonas aeruginosa was investigated in a rabbit model. Correlations between in-vivo CIP pharmacokinetics in peritoneal dialysate and serum after intraperitoneal administration, in-vivo cell counts and rabbit survival rate were obtained. Dialysate and serum concentrations after 12 h (C12h) were greater than those obtained with free drug whereas maximum serum concentrations (Cmax) were lower and the time to reach Cmax (tmax) was longer. A silastic implant device pre-colonised with P. aeruginosa for 2 days was implanted in the rabbit peritoneum, and dialysate with or without drug or microspheres was administered via a catheter. Rabbits receiving no antibiotic and those receiving free drug (10 mg in dialysate) died of peritonitis and septicaemia, whereas all rabbits given CIP microspheres recovered completely from infection. The viable count of P. aeruginosa was markedly reduced or eliminated from the catheter, the device and the peritoneal wall in CIP microsphere-treated rabbits but not in rabbits treated with free drug, as determined from histological and scanning electronmicroscopic evidence. These results demonstrate that sustained release of antibiotics at biofilm eradication concentrations (BEC) is required to treat biofilm infections associated with peritoneal implanted devices.
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Penicillin-resistant Streptococcus pneumoniae in Germany: Genetic relationship to clones from other European countries
SummaryPenicillin-resistant Streptococcus pneumoniae strains isolated in different parts of Germany between 1982 and 1992 were compared with penicillin-resistant isolates, mainly of serogroups 6, 9, 14, 19 and 23, from other European countries. The main clones were recognised by their serotypes, antibiotic resistance patterns and penicillin-binding protein properties, and this typing was confirmed by multi-locus enzyme electrophoresis for a sample of 43 selected isolates. Eleven of the 14 resistant German isolates could be assigned to five genotypes isolated also in other countries. These included representatives of two distinct serotype 23F lineages predominant in Spain and France; a cluster of three serotype 6B isolates identical to clones in Spain, France, Finland and Hungary; and a serotype 9V clone of a type prevalent in Spain and now also in France. Serotype 19A clones of the type found in Hungary were not collected in Germany. The data suggest that two 23F lineages, represented by seven isolates from different locations, have become disseminated in Germany. Several resistant types found in the former West Germany resembled those found elsewhere in Western Europe whereas those from East Germany were distinct or, in one case, resembled a clone from Hungary. These data may reflect pre-unification travel patterns.
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Susceptibility of Streptococcus pyogenes to azithromycin, clarithromycin, erythromycin and roxithromycin in vitro
More LessSummaryThe susceptibility of 180 clinical isolates of Streptococcus pyogenes from six regions of The Netherlands to the macrolide antibiotics azithromycin, clarithromycin, erythromycin and roxithromycin was analysed. The results of a microbroth MIC method, the E-test method and a disk diffusion assay were compared, and the MBC determined. In addition, the susceptibility to erythromycin of 436 clinical isolates of S. pyogenes from the Leiden region was determined. The microbroth MIC90s of azithromycin, clarithromycin, erythromycin and roxithromycin for group A streptococci were ≤ 0.5 mg/L. Erythromycin had the lowest MIC90 (0.09mg/L). The MIC data obtained with the E-test method suggested that clarithromycin and erythromycin had slightly higher anti-streptococcal activity than azithromycin and roxithromycin in vitro. MICs obtained with the E-test were lower than those found with the microbroth method. Only minor discrepancies were observed among the three methods. The MBC50 for both clarithromycin and erythromycin was 0.75 mg/L and 5.0mg/L for azithromycin and roxithromycin. None of the 180 strains and two of the collection of 436 strains (0.5%) were resistant to erythromycin and the other macrolides tested; MICs ranged from 1 to 16 mg/L. The erythromycin-resistant strains showed an inducible type of macrolide-lincosamide-streptogramin B (MLS) resistance.
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- Bacterial Characterisation
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Pheno-genotyping of verotoxin 2 (VT2)-producing Escherichia coli causing haemorrhagic colitis and haemolytic uraemic syndrome by direct analysis of patients’ stools
More LessSummaryThe subtype of verotoxin 2 (VT2) found in 22 VT2-positive stool samples from severely diseased Italian and German children with haemorrhagic colitis or haemolytic uraemic syndrome, or both, and that produced by the corresponding VT-producing Escherichia coli (VTEC) strains isolated from the stools were studied by cytotoxicity sero-neutralisation assays and by polymerase chain reaction (PCR) amplification of the VT2 B-subunit gene, followed by restriction fragment length polymorphism (RFLP) analysis. The free faecal toxin was serotyped as the classical VT2 in 21 stool samples, and as the VT2 variant VT2c in one. For all but one of the VTEC isolates, the toxin phenotype was consistent with the type of VT produced in vivo and found in the corresponding stool samples. Genotyping was in agreement with phenotyping for those strains harbouring a single type of VT2 gene. Three O157: H7 isolates carrying both VT2 and VT2c genes had the VT2 phenotype, instead of the expected VT2c phenotype. Direct PCR analysis of stools detected VT genes in only 11 of 20 VT-positive stool samples suggesting that the Vero cell cytotoxicity assay is more sensitive in diagnosing VTEC infection. Immunological and genetic subtyping of VT2 performed directly on stool samples from patients with haemolytic uraemic syndrome could be a useful complementary approach to understanding the role of the different types of VT in this syndrome.
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Cytotoxin detection in Campylobacter jejuni strains of human and animal origin with three tissue culture assay systems
More LessSummaryCytotoxin (CTX) production in 34 human and 22 animal strains of Campylobacter jejuni isolated in Japan and other countries was studied by three assay systems described previously. Furthermore, cholera-like enterotoxin production by these strains was tested by reversed passive latex agglutination (RPLA). CTX titres in the fetal calf serum (FCS) and newborn calf serum (NCS) assays were relatively lower, with a maximum of 4 and 8, respectively, than the maximum of 128 for the serum-free culture (SFC) assay. CTX detection rates were 62, 85 and 100% in human isolates and 64, 77 and 100% in animal isolates for the FCS, NCS and SFC assay systems, respectively. There was no significant difference in the detection rate of CTX between human and animal isolates, or between human isolates from Japan and other countries. With the three assay systems, the strains were divided into four groups from the pattern of CTX detection; 54% of strains gave positive results in all three assay systems, and 9 % of them were positive in the SFC assay only. Morphological changes on CHO cells showed distended instead of rounded cells with eight of 21 strains negative in the FCS assay. Cholera-like enterotoxin was not detected in the culture filtrate of any of the strains when tested by RPLA. These results indicate that cytotoxin production by C. jejuni is complex as compared with that of other enteric pathogens.
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- Immunological Response To Infection
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Antibody response to Staphylococcus aureus collagen binding protein in patients with S. aureus septicaemia and collagen binding properties of corresponding strains
More LessSummaryAn ELISA was developed for the detection of IgG antibodies to Staphylococcus aureus collagen binding protein (CnBP) in 95 patients with S. aureus endocarditis, complicated septicaemia with bone and joint involvement or uncomplicated septicaemia and in 95 control patients. Sixty percent of S. aureus-infected patients showed a positive peak anti-CnBP antibody level or a significant rise in titre, or both, during infection, but patients with S. aureus endocarditis or complicated septicaemia could not be differentiated from those with uncomplicated S. aureus septicaemia. The collagen binding capacity of S. aureus strains from 82 of the 95 patients was investigated in a particle agglutination assay and a 125 I-labelled assay. All strains bound collagen in the particle agglutination assay as did 68 % in the 125 I-labelled assay, but there was no correlation between collagen binding of the patient strain and endocarditis, joint or skeletal involvement. An anti-CnBP antibody response was seen in 16 patients infected with a S. aureus strain negative for collagen binding in vitro, indicating in-vivo expression of CnBP.
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Antibodies to meningococcal class 1 outer-membrane protein and its variable regions in patients with systemic meningococcal disease
More LessSummaryAntibodies to the meningococcal serosubtype-specific P1. 7, 16 protein and its variable regions (VR) were analysed in 28 convalescent sera drawn 8-36 months after systemic meningococcal disease by immunoblotting and enzyme immunoassay (EIA) methods. EIA antigens were the meningococcal P1. 7, 16 protein, produced in Bacillussubtilis, and peptides covering its VR1 (P1. 7 region) and VR2 (P1. 16 region) inserted into a bacterial penicillinase protein. In the immunoblotting method, three meningococcal reference strains were used; they expressed either the P1. 7, 16 protein, or only its VR1 or VR2 epitopes in their class 1 proteins. Both methods showed a strong IgG response in four sera to P1. 7, 16 and VR2, but not to VR1; 18 sera had no or weak anti-class 1 protein activity. The six remaining sera were positive only on blots. The VR2-specific sera had 30-fold higher bactericidal activity than those with negligible P1. 7, 16 responses. Previous vaccination of the patients with a B: 15: P1. 7, 16 meningococcal vaccine was associated with a strong anti-P1. 7, 16 and anti-VR2 booster response that declined with time. The subtype-specific antibody activity in some sera indicated colonisation after disease by meningococci with class 1 proteins different from the strain that had caused disease.
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Outer-membrane protein- and rough lipopolysaccharide-specific monoclonal antibodies protect mice against Brucella ovis
More LessSummaryBrucella ovis, a naturally virulent rough Brucella species, is the aetiological agent of ram epididymitis. The identification of protective antigens is necessary to obtain a safe, specific subcellular vaccine. Monoclonal antibodies (MAbs) directed at both brucella outer-membrane proteins (OMPs) and rough lipopolysaccharide (R-LPS) in a mouse protection test were used to identify potential targets for humoral immunity. Mixtures of MAbs directed at the 16.5-, 25–27-, 31-34- and 36-38-kDa OMPs conferred significant protection 7 days after challenge with reference strain B. ovis 63/290 compared with controls receiving either saline or an anti-brucella O-polysaccharide MAb. Furthermore, an anti-R-LPS MAb tested alone conferred protection at a level comparable with that obtained with the mixture of anti-OMP MAbs. The combination of protective OMP MAbs with the anti-R-LPS MAb was also strongly protective. One combination of OMP MAbs, which bound intensely to B. ovis in vitro, was ineffective. These results indicate that B. ovis OMPs and R-LPS are targets for protective antibodies and that they can be regarded as candidates for ram epididymitis subcellular vaccines.
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- Mycology
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An α5β1 -like integrin receptor mediates the binding of less pathogenic Candida species to fibronectin
More LessSummaryThe present study was undertaken to investigate whether less pathogenic Candida species (C. tropicalis, C. stellatoidea, C. krusei and C. glabratd) express a fibronectin receptor (FNr) antigenically related to α5βl integrin, which mediates their binding to fibronectin (FN). By flow cytometric analysis, a monoclonal antibody (MAb) directed against human α5 integrin subunit (clone SAM-1) and two different antisera to FNr positively stained C. tropicalis, C. stellatoidea and C. glabrata, with the greatest expression observed for C. tropicalis. No or only marginal immunoreactivity was found on C. krusei. C. tropicalis, C. stellatoidea, C. glabrata, but not C. krusei yeasts specifically adhered to FN; higher levels of adhesion were found for C. tropicalis and C. stellatoidea with respect to C. glabrata. Less pathogenic Candida spp. bound to the Arg-Gly-Asp (RGD) containing 120-kDa fragment of FN and adhesion to intact FN was markedly inhibited by Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP), but not by Gly-Arg-Gly-Glu-Ser-Pro (GRGESP) peptides. In addition, anti-α5 SAM-1 MAb and both anti-FNr antisera strongly blocked binding of less pathogenic Candida spp. to FN. Overall, these results indicate that less pathogenic Candida spp., including C. tropicalis, C. stellatoidea and C. glabrata, express a receptor antigenically related to α5βl integrin which mediates their adhesion to FN.
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- Icsb Announcement
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- Announcement
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- Book Received
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Volume 73 (2024)
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