In vitro bactericidal activity of cefpirome in combination with vancomycin against Staphylococcus aureus and coagulase-negative Staphylococcus.
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Anti-Bacterial Agents↗
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In vitro susceptibility of Staphylococcus aureus and coagulase-negative Staphylococcus strains to fusidic acid.
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Anti-Bacterial Agents↗
The pls gene found in methicillin-resistant Staphylococcus aureus strains is common in clinical isolates of Staphylococcus sciuri.
pls, a gene found in type I staphylococcal cassette chromosome mec (SCCmec) regions of methicillin-resistant Staphylococcus aureus strains, was present in 12 of the 15 human clinical Staphylococcus sciuri isolates studied. Pls was expressed in the S. sciuri isolates, although at a lower level than in S. aureus. Other parts of SCCmec could also be found in the S. sciuri genome.
Bacterial Proteins↗
Survey of staphylococcal enterotoxin genes, exfoliative toxin genes, and toxic shock syndrome toxin 1 gene in non-Staphylococcus aureus species.
The aim of this study was to analyze the presence of staphylococcal exotoxin genes in staphylococci other than Staphylococcus aureus. Whereas for Staphylococcus aureus a large spectrum of different exotoxins responsible for toxin-mediated diseases has been described, only few and conflicting data are available regarding toxin production in all other staphylococcal species. A collection of clinical non-Staphylococcus aureus staphylococcal isolates were systematically screened for the presence of genes for toxic shock syndrome toxin 1, exfoliative toxins, and classical enterotoxins by using multiplex polymerase chain reaction enzyme immunoassays. In a total of 461 isolates of 18 different (sub)species derived from clinical specimens, no toxin gene sequences were amplified. The results indicate that the occurrence of toxin genes involved in staphylococcal toxin-mediated diseases is at least very rare in human-derived isolates of non-Staphylococcus aureus species.
Bacterial Toxins↗
Molecular cloning and expression of a novel Staphylococcus aureus antigen.
A novel gene encoding an antigen from Staphylococcus aureus was isolated from an expression library by screening with antisera from patients with deep Staphylococcus aureus infections. In one positive clone an open reading frame, named ORF-2, was identified. Recombinant ORF-2 protein reacted with human immune serum. ORF-2 was shown to be present in other Staphylococcus aureus strains, but not in related species.
Amino Acid Sequence↗
Factors involved in the early pathogenesis of bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion. A review.
Staphylococcus aureus is the most important and prevalent contagious mammary pathogen; it causes clinical and subclinical intramammary infection with serious economic loss and herd management problems in dairy cows. In vitro studies have shown that Staphylococcus aureus adheres to mammary epithelial cells and extracellular matrix components and invades into mammary epithelial as well as other mammary cells. Staphylococcus aureus strains from intramammary infection produce several cell surface-associated and extracellular secretory products. The exact pathogenic roles of most of the products and their effects on adhesion and invasion are not well evaluated. It is also known that mammary epithelial cell-associated molecules and extracellular matrix components interact with S. aureus during the pathogenesis of mastitis, but their roles on adhesion and invasion have not been characterized. The adhesion of S. aureus to epithelial cells may involve non-specific physicochemical interactions and/or specific interactions between bacterial cell-associated ligands and host cell surface receptors. In vitro adhesion depends on the S. aureus strain, the growth phase of the bacteria, the growth medium and the origin of the epithelial cells. Adhesion is hypothesized to be a prerequisite and crucial early step for mammary gland infection. Staphylococcus aureus invades mammary epithelial cells. It also invades other cells such as endothelial cells and fibroblasts. Bacteria are found enclosed in membrane bound vacuoles in the cytoplasm of mammary epithelial cells. Recent observations indicate that S. aureus escapes from the phagosome into the cytoplasm and induces apoptosis. The invasion into mammary epithelial cells may occur through an endocytic process that requires involvement of elements of the cytoskeleton or by direct binding of bacteria to epithelial cells through a process mediated by specific receptors that needs de novo protein synthesis by both cells. Thus, the recurrent subclinical infection may result from this intracellular existence of bacteria that are protected from host defenses and effects of antibiotics. This review emphasizes on recent findings on S. aureus adhesion to mammary epithelial cells and extracellular matrix components and invasion into mammary epithelial cells.
Animals↗
Direct detection of human Staphylococcus aureus carriage in the nose using the Lightcycler Staphylococcus kit.
The Lightcycler Staphylococcus kit is a diagnostic tool for direct real-time detection of Staphylococcus aureus in clinical materials. We show here that detection of S. aureus nasal carriage using this test is hampered by competition of DNA from coagulase-negative staphylococci. However the test is well suited for species identification after culture and the identification of high-load S. aureus carriers.
Carrier State↗
Methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative Staphylococcus ocular surface infection efficacy of chloramphenicol eye drops.
PURPOSE: Methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococcus (MR-CNS) are two major multidrug-resistant pathogens. In this paper we report the prevalence of MRSA and MR-CNS in ocular surface infections. METHODS: We investigated the proportion of MRSA and MR-CNS in bacterial isolates from conjunctiva of elderly patients with and without bacterial conjunctivitis. The relationship between conjunctival MRSA carriers and general background conditions was studied. We evaluated the efficacy of chloramphenicol eye drops in the treatment of MRSA conjunctivitis. We also investigated the presence of MRSA and MR-CNS in lid skin and conjunctiva of patients with atopic dermatitis. RESULTS: In conjunctival bacterial flora of elderly patients the proportion of MRSA in S. aureus was 57%, and the proportion of MR-CNS in coagulase-negative Staphylococcus was 25%. Conjunctival MRSA carriers were more likely to have anemia, malignant tumor, liver dysfunction, and dementia, and to be postoperation and chronically bedridden. The efficacy rate of chloramphenicol eye drops in the treatment of MRSA conjunctivitis was 81%. In conjunctival sacs of patients with atopic dermatitis, S. aureus was the most frequent species (48%), and the proportion of MRSA was 18%. CONCLUSION: Methicillin-resistant S. aureus and MR-CNS are widespread in elderly hospitalized patients and in patients with atopic dermatitis. Chloramphenicol eye drops were useful for the treatment of MRSA ocular surface infections.
Aged↗
[The change in CD14 mRNA expression in rats with scald injury and Staphylococcus aureus challenge].
OBJECTIVE: To investigate the potential role and changes in CD14 in postburn Staphylococcus aureus sepsis. METHODS: Wistar rats were subjected to a 20% total body surface area (TBSA) full-thickness thermal injury and followed by Staphylococcus aureus challenge. Plasma and tissue samples from liver, kidneys, lungs and heart were collected to determine endotoxin (lipopolysaccharide, LPS) levels and CD14 mRNA expression. RESULTS: It was found that after thermal injury combined with Staphylococcus aureus sepsis, LPS levels in various tissues were markedly increased, peaking at 2 approximately 6 h (P < 0.05), then returned to baseline at 24 h. Plasma LPS level was also increased significantly at 2hrs after Staphylococcus aureus challenge compared with that in normal control animals (0.3056 EU/ml vs. 0.1250 EU/ml, P < 0.05). In addition, the intestinal diamine oxidase (DAO) activity decreased (P < 0.05). This indicated that the integrity of intestinal mucosa was damaged. Meanwhile, tissue CD14 mRNA expressions were up-regulated in all tissues (P < 0.05), especially in lungs. CONCLUSION: These data suggested that gut-originated LPS translocation and CD14 mRNA expression could be induced by thermal injury with Staphylococcus aureus sepsis. The up-regulation of CD14 gene expression might be associated with the local endotoxin activation secondary to postburn sepsis.
Animals↗
[Arbekacin resistant gene, aacA/aphD, in Staphylococcus aureus is lost during in vitro passage].
We analyzed for the presence of the aacA/aphD gene in arbekacin resistant-Staphylococcus aureus. Tested strains were 50 clinically isolated Staphylococcus aureus that had an MIC of more than 2 micrograms/ml for arbekacin. Among the primary cultures, aacA/aphD genes were detected in 42 of 50 strains(84%) overall, including 34 of 41 Methicillin resistant Staphylococcus aureus(MRSA) strains, and 8 of 9 strains of Methicillin sensitive Staphylococcus aureus(MSSA). After 10 or 20 passages, the aacA/aphD gene was lost in 5 strains of MRSA and 2 strains of MSSA. In the strains that lost the gene, the MIC of arbekacin was reduced remarkably in 3 strains of MRSA and 2 strains of MSSA. Further, MICs of arbekacin decreased along with passages even in the strains that did not loose the aacA/aphD gene, as well as in the strains that did not possess the aacA/aphD gene originally. The fact that MIC values are easily reduced with in vitro passage is important to consider when performing the arbekacin-resistance test for Staphylococcus aureus.
Aminoglycosides↗
Lipase versus teichoic acid and alpha-toxin as antigen in an enzyme immunoassay for serological diagnosis of Staphylococcus aureus infections.
Titres of IgG antibodies to Staphylococcus aureus lipase were analysed in 448 sera from patients suspected of having Staphylococcus aureus infections and the results compared to those for the routinely used staphylococcal antigens teichoic acid and alpha-toxin. The results indicated that determination of serum antibodies to lipase is a sensitive assay for serological diagnosis of staphylococcal infections and increased sensitivity may be achieved by selection of optimal antigen combinations.
Adolescent↗
Toll-like receptor 2 expression on human conjunctival epithelial cells: a pathway for Staphylococcus aureus involvement in chronic ocular proinflammatory responses.
BACKGROUND: Staphylococcus aureus colonization is common in atopic keratoconjunctivitis, potentially activating epithelial cells via toll-like receptor 2 (TLR-2) and the receptor for platelet-activating factor (PAFR). OBJECTIVES: To examine human conjunctival epithelial cells for the expression of TLR-2 in vitro and in vivo and to evaluate the role of TLR-2 in S aureus-mediated activation of these cells. METHODS: Conjunctival epithelial cells isolated from cadaveric tissues were stimulated with interferon gamma (IFN-gamma) or a commercial S aureus cell wall extract (Staphylococcus aureus-CWE) (with or without anti-TLR-2 blocking antibody or PAFR antagonist) and were analyzed for tumor necrosis factor alpha (TNF-alpha) and interleukin 8 (IL-8) release; surface expression of TLR-2, intercellular adhesion molecule-1, HLA, and CD14; and TLR-2 messenger RNA expression. Ocular surface cells collected via impression cytology were examined for TLR-2 expression via flow cytometry. RESULTS: Expression of TLR-2 was up-regulated on conjunctival epithelial cells by IFN-gamma and Staphylococcus aureus-CWE. Expression of TLR-2 messenger RNA was increased by IFN-gamma. Staphylococcus aureus-CWE up-regulated intercellular adhesion molecule 1, HLA, and CD14 expression and increased TNF-alpha and IL-8 release in a dose-dependent manner. Anti-TLR-2 significantly inhibited TNF-alpha release, whereas PAFR antagonist significantly inhibited IL-8 release. Toll-like receptor 2 was expressed on conjunctival epithelial cells from 4 of 5 patients with atopic keratoconjunctivitis, 3 of 5 with seasonal allergies, and 0 of 3 without allergies. CONCLUSIONS: Conjunctival epithelial cells express TLR-2 and may play an active role in the chronic ocular inflammatory response to S aureus through pathways that involve TLR-2 and PAFR.
Adolescent↗
Relation of cholesterol-stimulated Staphylococcus aureus growth to chronic blepharitis.
PURPOSE: Many types of chronic blepharitis have been believed to be primarily microbial in origin; however, it was proposed that differences and changes in lipid composition of meibomian secretion may be the initiating factor in some of these. It was recently reported that there are two subgroups of normals, those whose meibomian secretions contain high levels of cholesterol esters and those whose secretions contain very low levels of these esters. Thus, these subgroups of normals were defined on the basis of detailed lipid analyses of meibomian secretions from individuals showing no clinical signs of chronic blepharitis. All secretions from patients in the various disease groups contain high levels of these esters. Based on previous observations that in some chronic blepharitis disease groups certain Staphylococcus species were capable of hydrolyzing cholesterol esters, the authors tested the hypothesis that the resulting cholesterol might affect growth of Staphylococcus aureus. METHODS: Staphylococcus aureus growth stimulation in Mueller-Hinton broth by cholesterol was determined by colony forming units. Growth stimulation by cholesterol and other additives was also determined by the optical density 650 nm method. Statistical analyses included analysis of variance and the Student's t test. RESULTS: Cholesterol stimulated Staphylococcus aureus growth was significant during the first 24 hr period (20% increase at 25 microM cholesterol, P < 0.02), and for the total 48 hr period (40% increase at 400 microM cholesterol, P < 0.005) when compared to the respective control. Growth stimulation, determined by OD at 650 nm, in the presence of cholesterol was significantly greater (P < 0.02) than that in the presence of either sitosterol or cholestanol when the sterol concentration was 190 microM. CONCLUSION: These results suggest that the presence and hydrolysis of cholesterol esters of meibomian secretions may contribute to the proliferation of Staphylococcus spp, especially Staphylococcus aureus, observed in some chronic blepharitis disease groups.
Blepharitis↗
Contamination of room door handles by methicillin-sensitive/methicillin-resistant Staphylococcus aureus.
We investigated the contamination of room door handles by Staphylococcus aureus in wards of a university hospital. Door handles in 53 (27.0%) of 196 rooms were contaminated by methicillin-sensitive Staphylococcus aureus (MSSA) and/or methicillin-resistant Staphylococcus aureus (MRSA); MSSA was detected on door handles of 41 rooms (20.9%), MRSA on door handles of 17 rooms (8.7%), and MSSA and MRSA on the same door handles of five rooms (2.6%). The density of MSSA contamination was 1-2.6x10(4) colony forming units (cfu)/door handle, and that of MRSA was 1-6.0x10(3) cfu/door handle. The MRSA contamination rate on door handles of rooms with patients with MRSA was 19.0% (4/21 rooms) while that on door handles of rooms with patients without MRSA was 7.4% (13/175); the difference was not significant. These results suggest extensive contamination of MSSA and MRSA in the hospital environment.
Environmental Microbiology↗
[Identification of Staphylococcus aureus by the sero-inhibition of nuclease in blood culture broths].
Identification of Staphylococcus aureus in blood cultures by sero-inhibition nuclease test is described in this paper. No positive results were found in 392 Staphylococcus other than Staphylococcus aureus bacteremia. Only 0,8% Staphylococcus aureus bacteremia could not be detected. This sero-inhibition nuclease test provides a rapid, less than 3 hours, easy to perform, reliable method to identify Staphylococcus aureus in blood cultures. Results are obtained 24 hours earlier than the conventional methods.
Culture Media↗
Purification of a bone sialoprotein-binding protein from Staphylococcus aureus.
Bone sialoprotein (BSP) is selectively bound by Staphylococcus aureus cells isolated from patients suffering from infections of bone and joint tissues [Rydén C., Maxe, I., Franzén, A., Ljungh, A., Heinegård, D. & Rubin, K. (1987) Lancet II, 515]. We now report on the purification of a cell-wall protein from Staphylococcus aureus, strain O24, that possesses affinity for bone sialoprotein. Staphylococcal cell-wall components with capacity to inhibit binding of 125I-labeled BSP to staphylococcal cells were solubilized with LiCl (1.0 M, pH 5.0). Preparative SDS/PAGE and protein-overlay experiments revealed that inhibitory activity present in LiCl extracts resided in a fraction of polypeptides with M(r) 75,000-110,000. Staphylococcal proteins solubilized with LiCl were chromatographed on a Mono-Q anion-exchange column. Inhibitory activity was eluted at 0.6-0.8 M NaCl and could be further purified by affinity chromatography on BSP-Sepharose. Elution of the affinity matrix with 0.1 M glycine, pH 3.0, specifically eluted inhibitory activity. Analysis by SDS/PAGE revealed a single M(r) 97,000 polypeptide in the eluate. The purified M(r) 97,000 protein bound BSP in protein-overlay experiments. LiCl extracts from S. aureus, strain E514 or Staphylococcus epidermidis, strain 7686, both lacking the capacity to bind BSP did not contain the M9r) 97,000 protein. Our data demonstrate the presence of a S. aureus cell-surface BSP-binding protein. This protein could be involved in bacterial tropism in osteomyelitis.
Bacterial Proteins↗
In vitro and in vivo study of fosfomycin in methicillin-resistant Staphylococcus aureus septicaemia.
Five hundred strains of methicillin-resistant Staphylococcus aureus were tested against various anti-staphylococcal agents. Vancomycin, fusidic acid and fosfomycin were found to be the most effective. Only 1 strain out of 500 was resistant to fosfomycin. Three patients with methicillin-resistant Staphylococcus aureus septicaemia were successfully treated by fosfomycin. We conclude that fosfomycin could be the drug of choice for methicillin-resistant Staphylococcus aureus infection.
Adult↗
Failure of cephalosporins to prevent Staphylococcus aureus surgical wound infections.
Approximately 35,000 Staphylococcus aureus surgical wound infections occur annually in the United States. To investigate why S aureus causes infection despite the perioperative administration of cephalosporins, we compared 35 methicillin-susceptible isolates recovered from deep wound infections that complicated cefazolin prophylaxis (18 of 1650 patients) and cefamandole prophylaxis (17 of 3702 patients) with 64 colonizing isolates from presurgical patients. Compared with both colonizing and cefamandole-associated isolates, S aureus isolates from cefazolin-associated infections were more resistant to cefazolin by specialized assays. Staphylococcus aureus isolates that produced the A and C variants of staphylococcal beta-lactamase were associated with infections following cefazolin and cefamandole prophylaxis, respectively. These isolates hydrolyze the respective cephalosporins rapidly, suggesting that staphylococcal survival after perioperative prophylaxis may be mediated by in vivo degradation of the prophylactically administered cephalosporin. These data indicate that some S aureus wound infections occur because of deficiencies in antimicrobial effectiveness that are not detectable by routine susceptibility tests. This finding has important implications for the therapy and prevention of S aureus infection.
Cefamandole↗