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PubMed1999

Clinical features of Staphylococcus aureus endocarditis: a 10-year experience in Denmark.

BACKGROUND: Both morbidity and mortality resulting from Staphylococcus aureus endocarditis are known to be high, and the incidence of this disease seems to increase. The Statens Serum Institut, Copenhagen, Denmark, made it possible for us to analyze the clinical features of S aureus endocarditis in a nation-wide population of non-drug addicts. METHODS: Almost all Danish cases of bacteremia due to S aureus are reported to the Staphylococcus laboratory, Statens Serum Institut. The medical records were reviewed in cases reported from 1982 to 1991 in which the diagnosis of endocarditis was reported or suspected. RESULTS: A total of 260 patients, 145 males and 115 females, fulfilled the diagnostic criteria. The median age was 67.5 years. In 83 patients, the diagnosis of endocarditis was not suspected clinically. The overall mortality rate among those patients whose disease was diagnosed clinically was 46%. Among the subset of patients who received medical therapy only and appropriate antistaphylococcal treatment, mortality was significantly associated with late congestive heart failure, age, and involvement of the central nervous system. CONCLUSIONS: A raised awareness of the paucity of clinical findings and a more frequent use of echocardiography as a screening method seem essential to improve the prognosis of patients with S aureus endocarditis. Involvement of the central nervous system constitutes a relative indication of early valve replacement.

Adolescent↗

PubMed1981

Unusual multiresistant Staphylococcus aureus in a newborn nursery.

An apparently unique strain of Staphylococcus aureus, resistant to penicillins (including penicillinase-resistant penicillins), cephalosporins, cephamycins, aminoglycosides (all but amikacin), tetracycline, lincomycin, erythromycin, and, in a lesser percentage, to chloramphenicol as well, was isolated on more than 100 occasions over a 20-month period in a general hospital in San Sebastian, Spain. Forty-two (60%) of the 70 patients infected were patients in the newborn nursery. Many of these infants contracted serious disease. In the epidemiologic study of the nursery, this Staphylococcus strain was found in the air and on the clothing of the staff, although no nasal, perineal, or axillary carriers were found among the personnel.

Anti-Bacterial Agents↗

PubMed1984

Staphylococcus epidermidis sepsis in surgical patients.

We examined 58 surgical patients with two or more blood cultures positive for Staphylococcus epidermidis to determine factors associated with risk, mortality, and invasive sepsis. Bacterial sepsis was associated with gastrointestinal (GI) operations, total parenteral nutrition, and a regimen of two or more parenteral antibiotics. Mortality was 46% overall and significantly increased with age greater than 50 years, GI operations, other gram-positive sepsis, recurrent sepsis (positive blood culture 24 hours or more after the first blood culture), and the presence of organisms sensitive to three or less antibiotics. Antibiotic therapy appropriate for more than 50% of the S epidermidis organisms cultured from each patient resulted in significant reduction in mortality. Staphylococcus epidermidis should be considered a significant pathogen in critically ill surgical patients. Invasive S epidermidis sepsis can be recognized and requires specific antibiotic therapy.

Adolescent↗

PubMed1988

Immunomodulation by altered gastrointestinal tract flora. The effects of orally administered, killed Staphylococcus epidermidis, Candida, and Pseudomonas on systemic immune responses.

We investigated the role of microbial products from the gastrointestinal tract in the pathogenesis of immunosuppression in critical illness. Rats were gavaged daily for three weeks with killed Staphylococcus epidermidis, Candida, and Pseudomonas--organisms frequently isolated from the upper gastrointestinal tract of the critically ill patient; nonspecific immunity was measured by the delayed-type hypersensitivity (DTH) response. Gavage with either Pseudomonas or Candida resulted in significant suppression of DTH responses, while gavage with S epidermidis produced modest enhancement of DTH. Animals given Pseudomonas demonstrated impaired ability to localize a subcutaneous Staphylococcus aureus challenge and decreased IgM but enhanced anti-Pseudomonas IgA and IgG responses after intraperitoneal immunization. Pulmonary bacterial clearance at seven days was normal. These findings suggest that gut colonization with Candida or Pseudomonas may contribute to impairment of cell-mediated immunity in the critically ill patient.

Administration, Oral↗

PubMed1997

Staphylococcus epidermidis graft infection is associated with locally suppressed major histocompatibility complex class II and elevated MAC-1 expression.

OBJECTIVES: To determine the local cellular immune response in a series of human patients with Staphylococcus epidermidis prosthetic graft infection and to use a murine model to investigate the response in polytef (PTFE) and in a nonslime-producing S epidermidis variant. METHODS: Externally supported Dacron and PTFE grafts, either sterile or colonized with slime (RP-62A)- or nonslime (RP-62NA)-producing S epidermidis (10(7) colony forming units/cm2) were implanted in a dorsal subcutaneous pocket of Swiss Webster mice (Taconic, Germantown, NY). The grafts were harvested at 7, 10, 14, and 28 days with local bacterial and leukocyte counts obtained. Perigraft and blood monocyte major histocompatibility complex class II (MHC-II) (immune antigen) and membrane attack complex type 1 (MAC-1) (glycoprotein) expression were analyzed by flow cytometry in the murine model and in 3 patients representing 4 Dacron graft infections. RESULTS: The human infected Dacron perigraft monocytes revealed a suppressed MHC-II and elevated MAC-1 expression, and early correlation with the murine model was seen. No notable perigraft monocyte MHC-II suppression occurred in the infected PTFE graft. The reciprocal relationship in Dacron between monocyte MAC-1 and MHC-II expression was exaggerated with the lack of slime production. Bacterial clearance was variable. Supranormal expression was observed at 1 month in sterile Dacron but not in PTFE grafts. CONCLUSIONS: Staphylococcus epidermidis infection is associated with local cellular immune suppression in Dacron but not PTFE grafts. Slime-producing S epidermidis induced a lesser cytotoxic-phagocytic response than the nonslime variant. The reduced immunologic response to slime-producing S epidermidis may explain, in part, its indolent nature and resistance to eradication.

Animals↗

PubMed1978

Staphylococcus aureus endocarditis. Combined therapy with vancomycin and rifampin.

Two children with persistent bacteremia and endocarditis due to Staphylococcus aureus failed to respond to vancomycin therapy, even though serum levels greatly exceeded the inhibitory concentrations. The Staphylococcus from one patient was resistant to methicillin; the other patient had a penicillin hypersensitivity. There was a wide disparity between the minimum inhibitory and the minimum bactericidal concentrations of vancomycin. Striking clinical and laboratory evidence of improvement was demonstrated with the addition of rifampin therapy.

Blood↗

PubMed2004

Association between carriage of Streptococcus pneumoniae and Staphylococcus aureus in Children.

CONTEXT: Widespread pneumococcal conjugate vaccination may bring about epidemiologic changes in upper respiratory tract flora of children. Of particular significance may be an interaction between Streptococcus pneumoniae and Staphylococcus aureus, in view of the recent emergence of community-acquired methicillin-resistant S aureus. OBJECTIVE: To examine the prevalence and risk factors of carriage of S pneumoniae and S aureus in the prevaccination era in young children. DESIGN, SETTING, AND PATIENTS: Cross-sectional surveillance study of nasopharyngeal carriage of S pneumoniae and nasal carriage of S aureus by 790 children aged 40 months or younger seen at primary care clinics in central Israel during February 2002. MAIN OUTCOME MEASURES: Carriage rates of S pneumoniae (by serotype) and S aureus; risk factors associated with carriage of each pathogen. RESULTS: Among 790 children screened, 43% carried S pneumoniae and 10% carried S aureus. Staphylococcus aureus carriage among S pneumoniae carriers was 6.5% vs 12.9% in S pneumoniae noncarriers. Streptococcus pneumoniae carriage among S aureus carriers was 27.5% vs 44.8% in S aureus noncarriers. Only 2.8% carried both pathogens concomitantly vs 4.3% expected dual carriage (P =.03). Risk factors for S pneumoniae carriage (attending day care, having young siblings, and age older than 3 months) were negatively associated with S aureus carriage. CONCLUSIONS: Streptococcus pneumoniae carriage, specifically of vaccine-type strains, is negatively associated with S aureus carriage in children. The implications of these findings in the pneumococcal vaccine era require further investigation.

Carrier State↗

PubMed1986

The in vitro effects of methotrexate on the phagocytosis and intracellular killing of Staphylococcus aureus by human neutrophils.

The in vitro effect of therapeutic concentrations of methotrexate on the phagocytosis and intracellular killing of Staphylococcus aureus by circulating human neutrophils was assessed. Neutrophils were isolated from whole blood of six healthy human volunteers by density centrifugation and incubated with 10(-3) M methotrexate. Staphylococcus aureus was opsonized in human serum and added to the prepared neutrophils. Phagocytosis was determined by serial dilutions and plating of unphagocytized bacteria. After treatment with lysostaphin and trypsin, intracellular killing of bacteria was determined at set intervals by serial dilutions and platings of viable bacteria released by neutrophil lysis. Methotrexate-treated neutrophils phagocytized 56% and control neutrophils 67% of bacteria in 15 minutes (P greater than 0.5), and 96% of ingested bacteria were killed in 15 minutes by both populations. In these experiments, the in vitro incubation of circulating human neutrophils with methotrexate produced no significant alterations in neutrophilic phagocytosis or intracellular killing of S. aureus.

Humans↗

PubMed2001

Combinatorial synthesis through disulfide exchange: discovery of potent psammaplin A type antibacterial agents active against methicillin-resistant Staphylococcus aureus (MRSA).

Psammaplin A is a symmetrical bromotyrosine-derived disulfide natural product isolated from the Psammaplysilla sponge, which exhibits in vitro antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA). Inspired by the structure of this marine natural product, a combinatorial scrambling strategy for the construction of heterodimeric disulfide analogues was developed and applied to the construction of a 3828-membered library starting from 88 homodimeric disulfides. These psammaplin A analogues were screened directly against various gram positive bacterial strains leading to the discovery of a series of potent antibacterial agents active against methicillin-resistant Staphylococcus aureus (MRSA). Among the most active leads derived from these studies are compounds 104, 105, 113, 115, 123, and 128. The present, catalytically-induced, disulfide exchange strategy may be extendable to other types of building blocks bearing thiol groups facilitating the construction of diverse discovery-oriented combinatorial libraries.

Anti-Bacterial Agents↗

PubMed2002

Antibacterial activity of Australian plant extracts against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE).

Ethanolic extracts of five traditional Australian medicinal plants, previously shown to display antibacterial activity against laboratory strains of the Gram positive bacteria Staphylococcus aureus and Enterococcus faecalis, were investigated for their abilities to inhibit clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). Using plate-hole diffusion assays, the following results were obtained: (a) extract from the leaves of Eremophila alternifolia (Myoporaceae) showed activity against MRSA; (b) extract from the leaves of Acacia kempeana (Mimosaceae) showed incomplete inhibition of VRE; (c) extracts from the leaves of Amyema quandong (Loranthaceae) and Eremophila duttonii (Myoporaceae) were active against both types of bacteria; (d) extract from the stem base of Lepidosperma viscidum (Cyperaceae) was active against MRSA and exhibited incomplete inhibition of VRE. All active extracts were evaluated using time-kill assays. Most of the extracts showed bactericidal effects and reduced the number of viable cells by 4-6 logs within four hours, while the extracts from Acacia kempeana leaves and Lepidosperma viscidum stem base exhibited bacteriostatic activity against VRE. The extract from the leaves of Eremophila duttonii was the most active and reduced the number of viable cells of MRSA and VRE to undetectable levels within 1 hour.

Anti-Bacterial Agents↗

PubMed2001

Extracellular proteins of Staphylococcus aureus and the role of SarA and sigma B.

Staphylococcus aureus synthesizes a large number of extracellular proteins that have been postulated to play a role in bacterial virulence. The proteomic approach was used to analyse the pattern of extracellular proteins of two different S. aureus strains, RN6390 and COL. Thirty-nine protein spots were identified by N-terminal sequencing or MALDI-TOF-MS. The differences of the extracellular protein patterns between both strains are striking. Among the 18 proteins identified in S. aureus COL there are nine proteins not yet discovered in S. aureus RN6390. These are enterotoxin B, leukotoxin D, enterotoxin, serin proteases (SplA and SplC), thermonuclease, an IgG binding protein and two so far unknown proteins in S. aureus with similarities to SceD precursor in Staphylococcus carnosus and to synergohymenotropic toxin precursor in Streptococcus intermedius. In contrast, lipase as well as staphylokinase identified in S. aureus RN6390 were not detectable in S. aureus COL under the same conditions. By using a regulatory mutant of sarA (ALC136) isogenic to strain RN6390 we identified five proteins positively regulated by SarA and 12 proteins negatively regulated by SarA. Besides V8 protease (StsP) and Hlb already described to be regulated by the sar locus new putatively sarA-dependent proteins were identified, e.g. glycerolester hydrolase and autolysin both down-regulated in the sarA mutant, and aureolysin, staphylokinase, staphopain and format tetrahydrofolate lyase up-regulated in the mutant. Moreover, the role of sigma B in expression of extracellular proteins was studied. Interestingly, we found 11 proteins at an enhanced level in a sigB mutant of S. aureus COL, among them enterotoxin B, alpha and beta hemolysin, serine proteases SplA and SplB, leukotoxin D, and staphopain homologues. The sigma B-dependent repression of gene expression occurs at the transcriptional level. Only one protein, SceD, was identified whose synthesis was down-regulated in the mutant indicating that its gene belongs to the sigma B-dependent general stress regulon.

Amino Acid Sequence↗

PubMedDate not supplied

Enumeration of phagocytosed Staphylococcus aureus inside cells of the mouse macrophage cell line J774 by bioluminescence, fluorescence and viable counting techniques.

In order to quantify intracellular Staphylococcus aureus within a macrophage-like cell line by a bioluminescence technique, the mouse cell line J774 and opsonized Staphylococcus aureus were incubated together to allow phagocytosis to occur. Experiments using UV microscopy and fluorescent stained S. aureus were performed to determine an estimate of the mean intracellular bacterial numbers. For enumeration of intracellular bacteria by a bioluminescence technique, extracellular bacteria were removed by washing, the macrophages lysed mechanically and osmotically and treated with apyrase to remove somatic ATP. Bacterial cells were washed and the intracellular ATP measured by firefly luciferase bioluminescence in a luminometer. This new method of enumerating intracellular bacteria was compared to the conventional method of viable counts and found to correlate (r = 0.78). The bioluminescence assay developed was found to be a relatively rapid alternative method to the techniques currently used to enumerate intracellular bacteria and could prove advantageous in studies of intracellular killing and effects of antimicrobial agents on intracellular pathogens.

Animals↗

PubMed1999

Inhibition of cysteine protease and growth of Staphylococcus aureus V8 and poliovirus by phosphorylated cystatin alpha conjugate of skin.

The inhibitory properties of phosphorylated cystatin alpha (P-cystatin alpha) and a conjugated protein of the P-cystatin alpha with filaggrin linker segment peptide (FLSP) against the growth of Staphylococcus bacteria and poliovirus were investigated. Both the P-cystatin alpha and the conjugated protein (P-cystatin alpha-FLSP conjugate) as a model for the cornified envelope of skin inhibited the cysteine protease activity of Staphylococcus aureus V8. The protease activity was inhibited by normal cornified envelope of newborn rat skin, which contains P-cystatin alpha, and P-cystatin alpha in cornified envelope of newborn rat skin also suppressed the growth of S. aureus V8. When P-cystatin alpha or P-cystatin alpha-FLSP conjugate was added to cultured HeLa cells infected with poliovirus, 50-70% of the cell-death due to poliovirus infection was prevented. The poliovirus 3C protease activity in the infected HeLa cells was inhibited by P-cystatin alpha or P-cystatin alpha-FLSP conjugate. As a result, the processing of viral capsid peptides was suppressed. These findings suggest that P-cystatin alpha and P-cystatin alpha-FLSP conjugate could play the role of the barrier against microorganism infections due to inhibition of their cysteine protease activities.

Amino Acid Sequence↗

PubMed1975

Absorption of blocking activity from human tumor-bearer sera by Staphylococcus aureus, Cowan I.

Staphylococcus aureus, strain Cowan I, contains a cell-wall substance, protein A, which combines with the Fc part of IgG in most mammalian species. It can therefore be used as a solid-phase immunoabsorbant for elimination of the reacting immunoglobulins. Since it has been shown that Cowan I could absorb out the blocking activity of sera from rats bearing isografts of polyoma-virus-induced sarcomas or chemically induced colon carcinomas, we investigated what effects Cowan I absorption of human tumor-bearer sera might have. In all tumor-bearer sera tested, from patients with melanomas or colon carcinomas, treatment with protein-A-containing staphylococci decreased the sera's ability to inhibit lymphocyte-mediated cytotoxicity in vitro. Cowan-I-treated sera from healthy controls had no effect on lymphocyte cytotoxicity. Nor did Cowan-I-treated tumor-bearer sera potentiate or "arm" normal lymphocytes against tumor target cells. There was no evidence of complement-dependent cytotoxicity with added human complement in sera from melanoma and colon carcinoma bearing patients either before or after absorption with Staphylococcus aureus, Cowan I, The concentrations of IgA, IgG and IgM were determined in sera used for in vitro tests of blocking activity and complement-dependent cytotoxicity before and after absorption. No reduction of IgA, reduction to undetectable levels of IgG and 20-30 percent reduction of IgM immunoglobulins as compared to unabsorbed sera were demonstrated.

Absorption↗

PubMedDate not supplied

Effect of human serum on killing activity of vancomycin and teicoplanin against Staphylococcus aureus.

STUDY OBJECTIVE: To investigate the effects of pooled human serum (PHS) on the killing activity of vancomycin and teicoplanin against two isolates of Staphylococcus aureus from patients treated for endocarditis. DESIGN: An in vitro assessment of antibiotic susceptibility and killing rates. SETTING: An urban university teaching hospital. PATIENTS: Pooled human serum from patients treated for endocarditis. INTERVENTIONS: Two clinical isolates of Staphylococcus aureus were obtained from patients treated for endocarditis. Media consisted of cation-supplemented Mueller-Hinton broth alone and in 1:1 dilutions with PHS, 2-hour heat-inactivated PHS (HI-PHS), ultrafiltrate (UF), and 2-hour heat-inactivated ultrafiltrate (HI-UF). Heat inactivation of PHS and UF was accomplished by treatment at 56 degrees C for 2 hours. MEASUREMENTS AND MAIN RESULTS: Killing curves with vancomycin and teicoplanin were performed using drug concentrations of 45 micrograms/ml and a starting inoculum of approximately 1 x 10(6) colony-forming units (cfu)/ml. Bactericidal rates (-log cfu/ml/hr) were calculated from the slope of the killing curves over 0-12 hours (mean 3-8 replicates). CONCLUSIONS: The killing activity of vancomycin in PHS and HI-PHS against both isolates was significantly greater than all other media tested (p < 0.0001). Ultrafiltrate tended to reverse this enhancement effect. Addition of PHS or UF did not enhance teicoplanin's killing activity against either isolate. Further investigations in our laboratory will determine if the factor is antibiotic class or organism specific.

Blood Bactericidal Activity↗

PubMed1995

Bacteria/blood/material interactions. I. Injected and preseeded slime-forming Staphylococcus epidermidis in flowing blood with biomaterials.

Blood-material interactions were studied using in vitro recirculation with human blood, slime-forming Staphylococcus epidermidis, and cardiovascular materials. Staphylococcus epidermidis, under preseeded or injected conditions, adhered to nonsmooth materials and elevated plasma levels of fibrinopeptide A (FpA) and C3a in the presence of all materials. Increased white blood cell (WBC) and platelet adhesion and thrombospondin and platelet factor 4 (PF4) release were noted for respective materials in the presence of injected bacteria. Materials that adhered significant quantities of injected S. epidermidis exhibited low levels of adsorbed proteins. Materials with high levels of preseeded S. epidermidis showed high levels of adsorbed proteins. Adhesion of preseeded bacteria and blood plasma elevations of C3a and FpA were lowest on semicrystalline polymer substrates, intermediate on halogenated substrates, and highest on amorphous substrates. In the presence of injected bacteria, WBCs and platelets adhered at earlier recirculation times to amorphous substrates than to semicrystalline substrates.

Bacterial Adhesion↗

PubMed1984

Recruitment of neutrophils by an encapsulated coagulase-negative strain of Staphylococcus simulans in the mammary gland of the mouse.

The encapsulated coagulase-negative strain of Staphylococcus simulans (strain 76) was inoculated into the mammary glands of lactating mice. In contrast to the coagulase-positive encapsulated Staphylococcus aureus (strain M), which elicited a neutrophil response within 18 hr, strain 76 organisms multiplied in the glands but failed to elicit a neutrophil response for 3 days; they were then eliminated from the gland by 8 days. When strain 76 was inoculated into mice whose offspring had been removed 3 days earlier, a neutrophil response was induced within 18 hr and, although phagocytic ingestion was resisted for at least 42 hr, the organisms were eliminated from most glands within 5 days. Inoculation of a naturally occurring double-stranded RNA (BRL 5907) into the mammary gland induced a macrophage infiltration of the alveolar lumen that was qualitatively similar to 3 days involution. Intramammary inoculation of strain 76 18 hr after BRL 5907 resulted in a neutrophil infiltration within 6 hr. In vitro, strain 76 stimulated the release of chemotactic factors for neutrophils from bovine mammary gland macrophages. The results suggest that the recruitment of neutrophils by strain 76 in the mammary gland of the mouse is mediated by macrophages.

Animals↗

PubMed2007

Staphylococcus aureus peptidoglycan impairs fracture healing: an experimental study in rats.

Staphylococcus aureus is the common organism causing musculoskeletal infections. Staphylococcus aureus peptidoglycan (SaPG) has been identified to increase the acute inflammatory response to wounding, increase reparative granulation tissue, and improve healing. The healing of bone fractures is a balanced process of granulation tissue that is calcified to obtain increasing stability. By increasing reparative collagen accumulation, however, SaPG may induce a shift towards immature fibrous callus production. Therefore, it was our hypothesis that SaPG would impair bone healing after fracture. In three groups, each of nine rats, a mid-diaphyseal osteotomy/fracture of the femoral bone was performed and then nailed. In one group of animals, SaPG was applied locally at the fracture site, and in another group SaPG was applied intraperitoneally (systemically). Control littermate received saline. The animals were sacrificed after 6 weeks, and the mechanical characteristics of the healing osteotomies were evaluated. We found that application of SaPG locally induced a hypertrophic and immature callus as evaluated by callus production, by bone mineral content and density, and by bending moment and rigidity. In the rats given SaPG intraperitoneally, bone healing went uneventful compared to the control rats. Collectively, these data show that SaPG induces an alteration in the normal bone healing response towards a less calcified callus production.

Animals↗