J Bacteriol, 1998 Nov, 180(21), 5780 - 3 trans-complementation of a Staphylococcus aureus agr mutant by Staphylococcus lugdunensis agr RNAIII; Benito Y et al.; RNAIII from Staphylococcus lugdunensis (RNAIII-sl) in a Staphylococcus aureus agr mutant partially restored the Agr phenotype . A chimeric construct consisting of the 5' end of RNAIII-sl and the 3' end of RNAIII from S . aureus restored the Agr phenotype to a greater extent, suggesting the presence of independent regulatory domains. J Surg Res, 1998 Nov, 80(1), 28 - 34 Immobilized IgG and fibrinogen differentially affect the cytoskeletal organization and bactericidal function of adherent neutrophils; De La Cruz C et al.; The infection of a vascular prosthesis is a dreaded clinical outcome . Since fibrinogen (FBGN) and immunoglobulin (IgG) coat the implanted biomaterial surface, it is with these immobilized proteins that the neutrophil (PMN) interacts . This study tests the hypothesis that PMN are impaired in their ability to kill bacteria when bound to immobilized IgG or FBGN . Isolated human PMN were bound to FBGN or IgG and then left untreated or exposed to phorbol myristate acetate (PMA; 10(-7) M) . PMN adhered loosely, but did not spread, onto FBGN . In contrast, PMN spread fully onto IgG, exhibiting polarized pseudopodia . PMA treatment induced spreading of the FBGN bound cells . Suspended and adherent PMN were incubated 1 h with 14C-labeled Staphylococcus aureus; then, phagocytosis was assessed by radioactive uptake or bacterial kill was determined by plating recovered bacteria and colony counting . Data were analyzed by unpaired t test . We observed that both the phagocytic and the killing ability of FBGN-bound PMN were similar to that of suspended PMN . Conversely, IgG-bound PMN displayed a 62 +/- 6% (P < 0.01) decrease in phagocytosis and 33 +/- 7% (P < 0.05) reduction in kill vs suspended cells . PMA induced a 74 +/- 6% (P < 0.01) reduction in phagocytosis and 68 +/- 5% (P < 0.01) reduction in kill of bacteria for PMN bound to FBGN with no further effect on IgG-bound PMN . Using fluorescent vital dyes and confocal microscopy we determined that 33% fewer PMN were engaged in phagocytosis when bound to IgG vs FBGN . We conclude that Fc receptor ligation by immobilized IgG or PMA treatment of the FBGN-adherent PMN triggers cell spreading and reduced bactericidal activity . These results indicate that excessive cytoskeletal organization may impair the ability of PMN to kill bacteria and result in vascular graft infections . Proc Natl Acad Sci U S A, 1998 Oct 27, 95(22), 13125 - 9 Interleukin 12 deficiency associated with recurrent infections; Haraguchi S et al.; A 3-yr-old female patient exhibited interleukin 12 (IL-12) deficiency that was associated with recurrent episodes of pneumococcal pneumonia with sepsis and other infections in the absence of fevers . The patient's peripheral blood mononuclear cells (PBMCs) exhibited normal proliferative responses to antigens . Immune responses, including in vivo production of antibodies to diphtheria, tetanus, or pneumococcal antigens, were normal . Ig levels and B cell and T cell phenotypes were also normal . In contrast, IL-12 p70 heterodimer production was undetectable by using supernatants of the patient's stimulated PBMCs when compared with control cells treated similarly . Although present, interferon gamma (IFN-gamma) was reduced . The addition of recombinant IFN-gamma to control cells enhanced the production of IL-12 by up to sixfold . By contrast, IL-12 was undetectable in supernatants of the patient's cells in the presence of recombinant IFN-gamma . IL-12 p40 subunit mRNA by using the patient's PBMCs after stimulation with Staphylococcus aureus Cowan strain 1 or lipopolysaccharide was also undetectable by reverse transcription-PCR when compared with control cells . Production of IL-2, IL-6, tumor necrosis factor alpha, or IFN-gamma of the patient's PBMCs after appropriate stimulation was observed . This patient has either a defect in Staphylococcus aureus Cowan strain 1-lipopolysaccharide- or staphylococcal enterotoxin A-induced signaling pathways for the activation of IL-12 p40 gene expression, or an abnormality in the IL-12 p40 gene itself. Zentralbl Hyg Umweltmed, 1998 Sep, 201(3), 285 - 96 {Frequency and antibiotic susceptibility of oxacillin resistant Staphylococcus aureus in a teaching hospital}; Ritter E et al.; From October 1993 till October 1994, 115 oxacillin resistant Staphylococus aureus strains were isolated in the laboratory of a teaching hospital . This was 2.4% of all of the Staphylococcus aureus strains . The bacteria were isolated from 30 patients, 7 medical personnel and in the environment of the infected patients . Most of the isolates were cultured from blood cultures, wound swabs and drains . If the referring hospitals has been informed about the MRSA status of the patients, several transmissions could have been prevented . In 10 infected patients, the MRSA strains were isolated from the nose, throat and hands . The isolates were also found on the hands of several personnel and in the patients environment, suggesting that the strains had been widely spread . The MRSA strains predominated in the medical and surgical intensive care units and in 2 general surgical wards . They were only found sporadically in other departments (Ophthalmology, Gynaecology, Paediatrics and Urology) . MRSA-strains were more resistant to imipenem, ofloxacin, gentamicin, trimethoprim-sulfamethoxazole, tetracycline, erythromycin and clindamycin as oxacillin-sensitive Straphylococcus aureus strains . Genotyping (Restriction-Fragment-Length-Polymorphism) revealed six different strain patterns . The same RFLP types were mainly found on different wards . We conclude that various clones of MRSA may have emerged independently within one hospital and that their spread between wards was remarkably limited . Subsequent intensive hygiene measures have been successful in reducing the number of new isolates. J Clin Invest, 1998 Oct 15, 102(8), 1583 - 90 Antibacterial activity of human neutrophil defensins in experimental infections in mice is accompanied by increased leukocyte accumulation; Welling MM et al.; Neutrophil defensins (or human neutrophil peptides-HNP) are major constituents of the azurophilic granules of human neutrophils and have been shown to display broad-spectrum antimicrobial activity . Other activities of these defensins, which are released from stimulated neutrophils, include cytotoxic, stimulatory, and chemotactic activities toward a variety of target cells . We studied the potential use of HNP-1 for antibacterial therapy of experimental bacterial infections in mice . In experimental peritoneal Klebsiella pneumoniae infections in mice, HNP-1 injection was shown to markedly reduce bacterial numbers in the infected peritoneal cavity 24 h after infection . This antibacterial effect was found to be associated with an increased influx of macrophages, granulocytes, and lymphocytes into the peritoneal cavity . These leukocytes appeared to be a requirement for the antibacterial effect, since in leukocytopenic mice administration of HNP-1 did not display antibacterial activity . HNP-1 treatment also reduced bacterial numbers in experimental K . pneumoniae or Staphylococcus aureus thigh muscle infections . In this model, radiolabeled HNP-1 was found to accumulate at the site of infection, whereas most of the injected HNP-1 was rapidly removed from the circulation via renal excretion . These results demonstrate that neutrophil defensins display marked in vivo antibacterial activity in experimental infections in mice and that this activity appears to be mediated, at least in part, by local leukocyte accumulation. J Clin Invest, 1998 Oct 15, 102(8), 1473 - 80 Interactions between stromal cell--derived keratinocyte growth factor and epithelial transforming growth factor in immune-mediated crypt cell hyperplasia; Bajaj-Elliott M et al.; Immune reactions in the gut are associated with increased epithelial cell proliferation . Here we have studied the role of keratinocyte growth factor (KGF; FGF7) and transforming growth factor-alpha (TGF-alpha) in the epithelial cell hyperplasia seen in explants of fetal human small intestine after activation of lamina propria T cells with the superantigen Staphylococcus aureus enterotoxin B (SEB) . After the addition of SEB to the explants there is a 10-fold increase in KGF mRNA by 72 h of culture . KGF transcripts were abundant in the lamina propria using in situ hybridization and the culture supernatants contained elevated amounts of KGF protein . SEB had no direct effect on KGF mRNA and protein production by cultured lamina propria mesenchymal cells, but both were upregulated by TNF-alpha . Accompanying the increase in KGF there was also an increase in TGF-alpha precursor proteins in the culture supernatants and the phosphorylated form of the EGFR receptor was also detected in the tissue . Increased TGF-alpha precursor proteins were also detected in the supernatants of control explants stimulated with KGF alone . The direct addition of KGF and TGF-alpha enhanced epithelial cell proliferation and antibodies against KGF and TGF-alpha partially inhibited SEB-induced crypt hyperplasia . These results suggest molecular cross-talk between the KGF/KGFR and the TGF-alpha/EGFR in immune-mediated crypt cell hyperplasia. J Biol Chem, 1998 Oct 30, 273(44), 29143 - 9 Anchor structure of staphylococcal surface proteins . III . Role of the FemA, FemB, and FemX factors in anchoring surface proteins to the bacterial cell wall; Ton-That H et al.; Surface proteins of Staphylococcus aureus are covalently linked to the bacterial cell wall by a mechanism requiring a COOH-terminal sorting signal with a conserved LPXTG motif . Cleavage between the threonine and the glycine of the LPXTG motif liberates the carboxyl of threonine to form an amide bond with the pentaglycyl cross-bridge in the staphylococcal peptidoglycan . Here, we asked whether altered peptidoglycan cross-bridges interfere with the sorting reaction and investigated surface protein anchoring in staphylococcal fem mutants . S . aureus strains carrying mutations in the femA, femB, femAB, or the femAX genes synthesize altered cross-bridges, and each of these strains displayed decreased sorting activity . Characterization of cell wall anchor structures purified from the fem mutants revealed that surface proteins were linked to cross-bridges containing one, three, or five glycyl residues, but not to the epsilon-amino of lysyl in muropeptides without glycine . When tested in a femAB strain synthesizing cross-bridges with mono-, tri-, and pentaglycyl as well as tetraglycyl-monoseryl, surface proteins were found anchored mostly to the five-residue cross-bridges (pentaglycyl or tetraglycyl-monoseryl) . Thus, although wild-type peptidoglycan appears to be the preferred substrate for the sorting reaction, altered cell wall cross-bridges can be linked to the COOH-terminal end of surface proteins. J Biol Chem, 1998 Oct 30, 273(44), 29135 - 42 Anchor structure of staphylococcal surface proteins . II . Cooh-terminal structure of muramidase and amidase-solubilized surface protein; Navarre WW et al.; Surface proteins of the Gram-positive organism Staphylococcus aureus are anchored to the bacterial cell wall by a transpeptidation mechanism during which the polypeptide is cleaved between the threonine (T) and the glycine (G) of the LPXTG motif . The carboxyl of threonine is subsequently amide linked to the amino of the pentaglycyl cross-bridge within the staphylococcal peptidoglycan . Previous work examined the anchor structure of surface proteins solubilized from the peptidoglycan by treatment with lysostaphin or phi11 hydrolase and identified COOH-terminally linked triglycyl or L-Ala-D-iGln-L-Lys(Gly5)-D-Ala and MurNAc-{L-Ala-D-iGln-L-Lys(Gly5)-D-Ala}(beta1-4)-GlcNAc, respectively . Here, we report the anchor structure of surface proteins solubilized with N-acetylmuramidase and N-acetylmuramyl-L-alanine amidase . N-Acetylmuramidase-released surface protein was linked to MurNAc-{L-Ala-D-iGln-L-Lys(Gly5)-D-Ala}(beta1-4)-GlcNAc, whereas N-acetylmuramyl-L-alanine amidase treatment of the cell wall solubilized surface proteins linked to L-Ala-D-iGln-L-Lys(Gly5)-D-Ala . Most, but not all, anchor structures were cross-linked to other cell wall subunits, in which the D-alanyl at position four was amide linked to the pentaglycyl of a neighboring wall peptide. J Antimicrob Chemother, 1998 Sep, 42(3), 315 - 20 Increase in glutamine-non-amidated muropeptides in the peptidoglycan of vancomycin-resistant Staphylococcus aureus strain Mu50; Hanaki H et al.; The peptidoglycan compositions of vancomycin-resistant Staphylococcus aureus (VRSA) strain Mu50 (MIC 8 mg/L) and hetero-VRSA strain Mu3 (MIC 3 mg/L) were compared in order to understand the mechanism of vancomycin resistance . As compared with Mu3, the cell wall of Mu50 had increased amounts of glutamine-non-amidated muropeptides and decreased cross-linking of peptidoglycan with a greatly decreased dimer/monomer ratio of muropeptides . In agreement with this observation, the peptidoglycan of Mu50 bound 1.4 times more vancomycin than that of Mu3 . The increase in non-amidated muropeptides and the reduced cross-linking of the cell-wall peptidoglycan may contribute to the vancomycin resistance by increasing the consumption of vancomycin by the pre-existing cell wall of Mu50 and reducing the amount of vancomycin reaching the cytoplasmic membrane where the vital targets of the antibiotic are situated. N Z Med J, 1998 Sep 11, 111(1073), 345 - 7 Spinal epidural abscess; Wong D et al.; AIMS: To review the clinical presentation and outcome of patients with spinal epidural abscess . METHODS: Following an index case, additional cases were identified during 1991-6 . RESULTS: There were a total of seven patients with spinal epidural abscess and an average age of 52 years (range 24-75 years) . The abscess locations were cervical (3), thoracic (3) or thoracolumbar (1), and extended on average 4.3 vertebral bodies (range 2-9) . Staphylococcus aureus was the aetiologic agent in all of the six microbiologically confirmed cases . Three abscesses arose from adjacent vertebral osteomyelitis, one followed epidural anaesthesia and two arose by haematogenous spread . New spinal or radicular pain were the most frequent early symptoms, later, nerve root weakness or a sensory level . An ESR > 30 mm/hour was consistently present but fever and leukocytosis were absent in some patients . MRI (five cases) and myelography (one case) were diagnostic . Five patients underwent laminectomy and abscess drainage; in three, limb weakness improved markedly post operatively . Three of the four patients with paralysis died, two despite laminectomy . CONCLUSIONS: New spinal pain, radicular symptoms or signs, and a raised ESR were the most consistent early abnormalities in patients with a spinal epidural abscess . Diagnosis at an early clinical stage was associated with a better outcome. Microbios, 1998, 94(378), 95 - 102 Attenuated mutants of Staphylococcus aureus with two temperature-sensitive lesions: isolation and characterization; Sordelli DO et al.; The feasibility of constructing attenuated mutants of Staphylococcus aureus with two temperature-sensitive (ts) lesions for ultimate development of a live-attenuated strain was investigated . Temperature-sensitive S . aureus strain G/1/2, which grows well at 31 degrees C but does not replicate at 37 degrees C, was subjected to chemical mutagenesis . After two enrichment cycles, fifteen mutants able to grow at 25 degrees C but unable to grow at 31 degrees C, were identified . Growth curves with temperature shifts from 25 to 31 degrees C, and from 31 to 37 degrees C confirmed that these were mutants with two lesions (dts), each with a different cut-off temperature . The reversion frequency of mutant G/1/2 at 37 degrees C was 2 x 10(-6) whereas those of several dts mutants were much lower (dts7: 7 x 10(-9) and dts12: 1 x 10(-9)) . There was no increase in ts mutation reversion rate in response to prolonged incubation at 37 degrees C . The data support the further development of these mutants for use as a stable attenuated vaccine. Infect Immun, 1998 Nov, 66(11), 5183 - 9 Staphylococcus aureus serotype 5 capsular polysaccharide is antiphagocytic and enhances bacterial virulence in a murine bacteremia model; Thakker M et al.; Controversy persists over the role that the capsular polysaccharide plays in the pathogenesis of Staphylococcus aureus infections . To address this issue, we compared the mouse virulence of S . aureus Reynolds and capsule-defective mutant strains cultivated under conditions of high or low capsule expression . Strain Reynolds cells cultivated on Columbia salt agar plates expressed approximately 100-fold more type 5 capsular polysaccharide than did cells cultivated in Columbia salt broth . The relative virulence of strain Reynolds and its capsule-defective mutants after growth on either solid or liquid medium was examined in mice challenged intraperitoneally or intravenously . The results indicated that agar-grown Reynolds cells were cleared from the bloodstream of mice less readily than broth-grown Reynolds cells . When the parental and mutant strains were cultivated on solid medium, strain Reynolds sustained a higher level of bacteremia than did the capsular mutants . We performed in vitro opsonophagocytic killing assays to determine whether staphylococcal virulence for mice correlated with resistance to phagocytosis . S . aureus Reynolds cultivated on solid medium was susceptible to phagocytic killing only in the presence of specific capsular antibodies and complement . Strain Reynolds grown in broth showed opsonic requirements for phagocytic killing that were similar to those of the capsular mutants (grown in broth or on agar); i.e., the bacteria were opsonized for phagocytosis by nonimmune serum with complement activity . These studies indicate that optimal expression of capsule enhances bacterial virulence in the mouse model of bacteremia, probably by rendering the organisms resistant to opsonophagocytic killing by leukocytes. J Nat Toxins, 1998 Oct, 7(3), 193 - 213 Induction of type 2 cytokines by a staphylococcal enterotoxin superantigen; Ferens WA et al.; Persistent intramammary infections of dairy cows with Staphylococcus aureus may involve immunosuppression mediated by bacterial toxins such as enterotoxins and other super-antigens (SAgs) . Previously we found that stimulation of bovine PBMC with staphylococcal enterotoxin C (SEC) induced a unique phenotype of activated CD8+ T cells expressing a newly identified activation molecule, ACT3 . In the present study we found that SEC induced the expression of interleukin (IL)-4 and IL-10 mRNAs, two cytokines associated with type 2 responses . Elevated levels of IL-4 and IL-10, observed between day 0 and day 4 of culture, were associated with temporary inhibition of proliferative responses of T cells, evidenced by a decrease in numbers of CD4+ T cells and a small increase in numbers of CD8+ T cells . Vigorous proliferation of T cells occurred between days 4 and 7 of culture and with a bias towards CD8+ T cells . Acquisition of the ACT3+ phenotype by CD8+ T cells was preceded by induction of IL-4 mRNA . Thus, in the bovine system, SAgs may hinder protective responses by inducing type 2 cytokines, which interfere with immune clearance of many microbial pathogens . The results of the study are consistent with the hypothesis that SAgs are involved in immunosuppression, and suggest possible immunomodulatory mechanisms. Eur Arch Otorhinolaryngol, 1998, 255(7), 347 - 51 A molecular epidemiologic study of methicillin-resistant Staphylococcus aureus infection in patients undergoing middle ear surgery; Suh HK et al.; The incidence of methicillin-resistant Staphylococcus aureus (MRSA) infections after middle ear surgery has recently increased at our hospital . Most of these infections were thought to be hospital-acquired when medical personnel in contact with an MRSA-infected patient may have inadvertently transmitted the pathogen to other patients . To prevent further transmission it is essential that such sources of MRSA infection and transmission routes be selected out and eradicated . Therefore, it is necessary to determine whether the strains of MRSA isolated from infected patients are identical to those obtained from medical personnel in order to prove a reciprocal transmission of organisms between medical personnel and patients . Surveillance bacterial cultures from the anterior nares and hands of medical personnel working in the Department of Otolaryngology, Korea University Guro Hospital, were performed at two different time points: 6 December 1994 and 17 June 1996 . Ribotyping with Southern blot technique was used to compare 12 MRSA strains from medical carriers with 60 strains identified from the otorrhea of MRSA-infected patients undergoing middle ear surgery . As results, six different MRSA strains were identified (types I, II, III, IV, V and VI) from ribotyping with EcoR1 . One distinct subtype, type I strain, was the most frequently identified strain in both medical carriers and patients . Results also showed that 6 MRSA isolates from 10 medical carriers and 20 from 30 patients contained type I ribotype at first culture . Two medical carriers' isolates and 13 isolates from 30 patients shared the same type I strain at the second surveillance culture . In all, 41 out of 72 MRSA strains (56.9%) shared an identical ribotype pattern . Postoperative MRSA infection rates after treatment of medical carriers and the application of rigorous preventive procedures decreased from 11.9 to 5.7% after first culture and 9.0 to 7.7% following second cultures . These findings confirm that MRSA transmission can occur between medical personnel and patients and that effective preventive measures can reduce the postoperative infection rate. Ostomy Wound Manage, 1998 Aug, 44(8), 32 - 40; discussion 34-8; quiz 41-2 Bacteria for the nineties; Devlin HR; Staphylococcus aureus and Enterococci have gained prominence as the causes of wound infections during this decade . Methicillin-resistant Staphylococcus aureus (MRSA) became commonplace in the United States during the 1980s . In Canada, infections with MRSA have been increasing in frequency since 1995 . MRSA develops resistance by producing an altered penicillin-binding protein, PBP 2a, coded for by the mecA gene . Vancomycin is the usual drug of choice . Recently, strains with intermediate resistance to vancomycin (VISA) have been isolated from patients in Japan and the United States . Interim guidelines for their control have been developed by the Centers for Disease Control . Enterococci have developed a resistance to a variety of antimicrobials during the past three decades, including beta-lactams and aminoglycosides . Recently, strains resistant to vancomycin (VRE) have been found in the United States and Canada . They are particularly difficult to treat, although some success has been achieved with experimental drugs . These microorganisms have the ability to escape control by antimicrobials almost as soon as they are developed . Thus, we must practice good infection control and reserve antimicrobials only for clear cases of infection if we are to prevent or delay the emergence of resistance. Microbiology, 1998 Sep, 144 ( Pt 9), 2469 - 79 The role of environmental factors in the regulation of virulence-determinant expression in Staphylococcus aureus 8325-4; Chan PF et al.; Staphylococcus aureus is a major human pathogen, which produces a variety of virulence determinants . To study environmental regulation of virulence-determinant production, several transcriptional reporter gene fusions were constructed . Chromosomal fusions were made with the staphylococcal accessory regulator (sarA), alpha-haemolysin (hla), surface protein A (spa) and toxic-shock syndrome toxin-1 (tst) genes . The effect of many different environmental conditions on the expression of the fusions was examined . Expression of hla, tst and spa was strongly repressed in the presence of sodium chloride (1 M) or sucrose (20 mM), but sarA was relatively unaffected . The global regulator of expression of virulence-determinant genes, agr (accessory gene regulator) was not involved in the salt or sucrose repression . Novobiocin, a DNA gyrase inhibitor, did not significantly increase the expression of tst in wild-type or agr backgrounds and failed to relieve the salt suppression . Expression of tst was strongly stimulated in several low-metal environments, independently of agr, whilst spa levels were significantly reduced by EGTA . The complex, interactive role of environmental factors in the control of expression of the virulence determinants is discussed. Chirurg, 1998 Aug, 69(8), 872 - 6 {The infected arterial stent}; Muller G et al.; Vascular infections after percutaneous endovascular procedures are rare . We report a case of bacterial arteritis following PTA and stent implantation into an iliac artery . It was complicated by septic embolization, ipsilateral empyema of the knee joint and formation of an infected false aneurysm . Contamination of the stent with Staphylococcus aureus is likely to be the cause . The treatment included resection of the infected arterial segment, removal of the stent, and autologous reconstruction. Chirurg, 1998 Aug, 69(8), 801 - 5 {Methicillin-resistant Staphylococcus aureus (MRSA)--clinical implications}; Peltroche-Llacsahuanga H et al.; The isolation rate of MRSA from Staphylococcus aureus infections rose to 8.7% in German hospitals between 1990 and 1995 . Patients undergoing surgical treatment or physiotherapy showed a threefold increase in the risk of being colonized or infected with MRSA . Surgical wound infection is the most frequent among MRSA-induced infections (28%) . The main transmission path of MRSA inside a hospital is bacterial spread from one patient to another through contact with the hands of nursing staff . Therefore, the crucial strategy in avoiding the spread of bacteria is strict hygiene management through hand disinfection . The most widespread therapeutic regimen for simultaneous eradication of nasal colonization and treatment of infection on other body sites is mupirocin nasal ointment combined with parenteral vancomycin application . The non-indicated use of vancomycin, e.g., for perioperative prophylaxis or prevention of catheter-induced infections, should be avoided, especially after the appearance of vancomycin-intermediately sensitive S . aureus (VISA) strains that have been reported recently from Japan and the USA. J Orthop Trauma, 1998 Sep-Oct, 12(7), 479 - 84 High pressure pulsatile lavage of contaminated human tibiae: an in vitro study; Bhandari M et al.; OBJECTIVE: This study was designed to examine the effect of high pressure pulsatile lavage (HPPL) on bone destruction and propagation of bacteria in experimentally contaminated human tibiae . METHODS: Using an in vitro model, nine human tibiae from above-knee amputations were tested . A mid-diaphyseal tibial shaft fracture was created, and each end of the fracture was contaminated with bacteria (six tibiae with Staphylococcus aureus, three tibiae with Escherichia coli) . The proximal end was designated as the control and the distal end was the test site . The test site was debrided by HPPL (seventy pounds/square inch, 1,200 milliliters/minute, 1,050 cycles/minute) with three liters of normal saline, whereas the control site did not receive any form of irrigation . Serial sections at increasing distance from the fracture site were cultured and the numbers of bacterial colony-forming units (CFUs) were determined at each level . The degree of macroscopic architectural change in each serial section was graded on an ordinal scale . RESULTS: Analysis of culture data revealed a reproducible pattern of bacterial propagation into the intramedullary canal . Peak bacterial seeding occurred at two to three centimeters from the fracture site (p = 0.023, Wilcoxon signed rank test) . The degree of bone destruction varied proportionally with the depth into the canal and was found to be predictive of the extent of bacterial propagation determined by culture data . CONCLUSION: In an in vitro model of a contaminated fracture, HPPL resulted in bacterial seeding into the intramedullary canal and significant damage to the architecture of the bone . These observations might have clinical significance. FEBS Lett, 1998 Oct 2, 436(2), 202 - 8 Characterization of a novel structural member, LukE-LukD, of the bi-component staphylococcal leucotoxins family; Gravet A et al.; A new member of the staphylococcal bi-component leucotoxins family, LukE (32 kDa) and LukD (34.3 kDa) has been characterized from Staphylococcus aureus strain Newman . LukE was 58-68% identical with the class S proteins, whereas LukD was 71-77% identical with the class F proteins of the family . A partial immunoreactivity with the various affinity-purified antibodies specific for the other proteins was observed . Immunoprecipitation assay and gene probing confirmed a 30% frequency among human clinical isolates, differing from the distribution of the other known leucotoxins (P<0.005) . LukE+LukD was as effective as the Panton-Valentine leucocidin for inducing dermonecrosis when injected in the rabbit skin, but not hemolytic and poorly leucotoxic compared to other leucotoxins expressed by Staphylococcus aureus. Adv Exp Med Biol, 1998, 443, 321 - 30 Effect of lactoferrin on the phagocytic activity of polymorphonuclear leucocytes isolated from blood of patients with autoimmune diseases and Staphylococcus aureus allergy; Manev V et al.; Phagocytic number (PN) and phagocytic index (PI) of neutrophils isolated from blood of patients with autoimmune diseases, allergy to Staphylococcus aureus and from blood of healthy individuals were examined . Our results concerning the influence of lactoferrin (Lf); (6.7 mg/l) on the PI of PMN showed that: 1) Lf enhances reliable PI of PMN at the 30-th minute starting the phagocytic reaction in patients with autoimmune disease in an active stage, in blood donors treated as healthy with the presence of autoantibodies, in patients with autoimmune diseases and proved autoantibodies against tissue, cell antigens and collagen, 2) Lf influences non-significantly PI of PMN in patients with autoimmune collagen diseases in remission, 3) Lf increases PI of PMN with 19% only in 58% from the assessed patients with Staphylococcus aureus, and 4) Lf decreases non-significantly PI of PMN in the healthy controls . Our studies on the effect of Lf on the phagocytic activity of PMN suggest that Lf has stronger effect on the PN compared to the PI: 1) Lf enhances with 86% the PN in patients with Staphylococcus aureus, 2) Lf increases PN of PMN in all of the assessed patients with autoimmune collagen diseases in active stage (mean with 72%), and 3) Lf increases PN of PMN in 4 from the 5 investigated healthy controls (mean with 22%) . Our results show a "corrective" effect of Lf on the phagocytic functions in the investigated groups of patients . The possible mechanisms, by which Lf increases PN and PI of neutrophils, is discussed: 1) they may concern the antioxidative properties of Lf to block the iron ions in their catalytic inactive form or to take part as ferric-Lf in an oxidative-reduction processes on the plasma membrane and controlling transmembrane transport systems, 2) Lf decreases the negative surface charge and thus enhances the adherent ability of the PMN . Probably to this stimulated adherent ability dues the increased ingestion of bacteria in the presence of Lf, and 3) The "changed" membrane of PMN may have higher number receptors for Lf to bind more molecules of exogenous Lf . The increase of Lf binding which enhances the adherence and aggregation of neutrophils, facilitates the phagocytosis. Rev Prat, 1998 Mar 1, 48(5), 523 - 7 {Surgery of infectious endocarditis}; Gandjbakhch I et al.; Thirty to fifty percent of patients with infective endocarditis are operated on during the active phase of the disease; this percentage is higher in case of some valvular localizations (aortic), in case of early prosthetic valve endocarditis, in case of some microorganisms (Staphylococcus aureus, gram-negative, fungus, intracellular microorganism) . Operative death (at 30 days) is below 10% in native valve endocarditis, close to 50% in early prosthetic valve endocarditis, and below 20% in late prosthetic valve endocarditis . When active infective disease has been healed by medical treatment alone, half the patients need surgery in the first 2 years of follow-up; the indications for surgery are the functional status, the degree of valvular leaks and other lesions, the degree of ventricular dilatation. Yeast, 1998 Sep 15, 14(12), 1139 - 46 New constructs and strategies for efficient PCR-based gene manipulations in yeast; Puig O et al.; Gene disruption and tagging can be achieved by homologous recombination in the yeast genome . Several PCR-based methods have been described towards this end . However these strategies are often limited in their applications and/or their efficiencies and may be technically demanding . Here we describe two plasmids for C-terminal tagging of proteins with the IgG binding domain of the Staphylococcus aureus protein A . We also present simple and reliable strategies based on PCR to promote efficient integration of exogenous DNA into the yeast genome . These simple methods are not limited to specific strains or markers and can be used for any application requiring homologous recombination such as gene disruption and epitope tagging . These strategies can be used for consecutive introduction of various constructs into a single yeast strain. Mol Biotechnol, 1998 Aug, 10(1), 9 - 16 A study of the interactions between an IgG-binding domain based on the B domain of staphylococcal protein A and rabbit IgG; Brown NL et al.; The nonantigenic interaction between a recombinant immunoglobulin G (IgG)-binding protein based on the B domain of Protein A from Staphylococcus aureus (termed SpA1) and the Fc fragment of rabbit IgG has been investigated . The contribution to binding of four putative hydrogen bond contacts between SpA1 and IgG-Fc were examined by the individual substitution of the residues in SpA1 involved in these interactions by others unable to form hydrogen bonds . It was found that the most important of the hydrogen bonds involved Tyr 18 which, when replaced by Phe, resulted in a twofold decrease in IgG-binding affinity . The residues of SpA1 proposed to make close, mainly hydrophobic, contacts with Fc were replaced by residues with potential electrostatic charge to establish the importance of the hydrophobic interaction in the complex . The IgG-binding affinities of the mutant proteins were compared to the wild-type protein by a competitive enzyme-linked immunosorbent assay . The replacement of individual hydrophobic residues by His generated a number of novel IgG-binding proteins with reduced binding affinity at pH 5.0 but which maintained strong binding affinities at pH 8.0 . The elution profile of human IgG1-Fc (Fc fragment of human IgG1) from a column made from an immobilized two-domain mutant protein shows that the complex dissociates at a higher pH relative to that of the non-mutated protein thus offering favorable elution characteristics. Infect Dis Clin North Am, 1998 Sep, 12(3), 613 - 30, viii Resistance issues and treatment implications: pneumococcus, Staphylococcus aureus, and gram-negative rods; Low DE; During the last decade there has been an unexpectedly rapid evolution of antimicrobial resistance in the respiratory pathogens for community- and hospital-acquired pneumonia . In order to choose the most optimal therapy for their patients, it is essential that physicians be aware of the prevalence and mechanisms of resistance and their implications on the effectiveness of the various antimicrobials. Can J Cardiol, 1998 Sep, 14(9), 1148 - 50 Staphylococcus aureus pericarditis with tamponade complicating coronary angioplasty and stenting; Amin H et al.; Infectious complications of coronary angioplasty and stenting are uncommon . A 70-year-old man is presented who underwent percutaneous transluminal coronary angioplasty (PTCA) and stenting of an occluded left anterior descending artery . This was complicated by Staphylococcus aureus pericarditis with tamponade . He was successfully treated with a closed drainage and antibiotics . This is the first reported case in the literature that documents purulent pericarditis and tamponade following percutaneous revascularization. Infect Control Hosp Epidemiol, 1998 Sep, 19(9), 643 - 6 Central venous catheters as a source of hemodialysis-related bacteremia; Taylor GD et al.; OBJECTIVE: To describe investigations into an increase in hemodialysis-related bacteremia that occurred in our hospital in the first 6 months of 1996 . SETTING: Hemodialysis unit in a tertiary-care medical center . METHODS: Prospective surveillance for hemodialysis bacteremia has been performed for several years . Cases that occurred in 1995 were compared to cases in the first 6 months of 1996 . Unit data on dialysis runs and method of dialysis access were used to calculate rates . Nested polymerase chain reaction (PCR) was used to type 18 Staphylococcus aureus isolates from 1996 . A case-control study comparing 80 randomly selected hemodialysis patients from 1995 and 1996 was performed to examine infection risk factors . RESULTS: The hemodialysis bacteremia rate was 1.2 per 1,000 runs in 1995 and 2.8 per 1,000 in the first 6 months of 1996 (P=.0009) . The 25 cases in 1995 and 32 in the first half of 1996 were similar in age, gender, means of vascular access, and microbial etiology . Central venous catheter (CVC) access accounted for >90% of cases in both time periods . S aureus was the most common microbial etiology (53% of the 1996 cases) . PCR typing of S aureus isolates from 1996 demonstrated five different strains, the most common having six isolates . The use of CVCs as a means of vascular access abruptly increased in the unit in January 1996, from <30% of dialysis runs in 1995 to >40% in 1996 (P<.001), associated with structural changes in healthcare delivery in the region resulting in delays in performing surgical procedures, such as creation of vascular grafts and fistulae . CONCLUSION: A marked increase in hemodialysis bacteremia occurred in 1996, associated with increased reliance on CVCs for vascular access in hemodialysis patients during a period of healthcare restructuring. J Hosp Infect, 1998 Sep, 40 Suppl B, S39 - 44 New strategies for the use of mupirocin for the prevention of serious infection; Mehtar S; Nasal mupirocin has an important role to play in the prevention of Staphylococcus aureus infection by eliminating nasal carriage of this organism . Indeed, in many countries nasal mupirocin is one of the mainstays for controlling outbreaks of methicillin-resistant S . aureus . Eradication of nasal S . aureus with mupirocin has been shown to be effective in preventing postoperative infections in patients undergoing cardiothoracic surgery and in preventing exit-site infections in patients undergoing haemodialysis . It has been proposed that the use of mupirocin should be extended to other situations, such as the prevention of postoperative infections in patients undergoing implant surgery and the prevention of bacteraemias in high-risk patients . Clinical trials are needed to establish the efficacy of mupirocin in these situations . Both low-level and high-level resistance have been reported during treatment with nasal mupirocin . Low-level resistance does not represent a significant clinical problem but high-level resistance resulting from indiscriminate use may give grounds for concern . Further review of these issues is required . As with any antibiotic, mupirocin should be used judiciously, as part of an integrated programme of infection control. J Hosp Infect, 1998 Sep, 40 Suppl B, S31 - 7 Eradication of nasal carriage of Staphylococcus aureus--is it cost-effective? Davey P. In cardiothoracic surgery, the costs of surgical-site infection (SSI) arise from additional postoperative procedures (approximately US $5000 per patient) and prolonged hospital stay (approximately $11,500 per patient) . Application of nasal mupirocin reduced SSIs by 63% compared with historical controls . This would have resulted in savings provided that the attributable cost of an SSI was more than $245 . Mupirocin was estimated to reduce the risk of bacteraemia in haemodialysis patients by 84% compared with historical controls . A model using data on Medicare payments for haemodialysis admissions was used to estimate the impact on hospital costs . The conclusion was that mupirocin would have been cost-saving but the model did not provide sufficient detail about hospital costing to allow assessment of its relevance in other settings . In a prospective, randomized, placebo-controlled trial in continuous ambulatory peritoneal dialysis (CAPD) patients, mupirocin reduced the risk of staphylococcal exit-site infection (ESI) from 0.42 to 0.14 per patient-year . However, as in a previous comparison with historical controls, there was an increase in the rates of ESIs caused by Gram-negative bacteria in patients who received mupirocin, bringing the rate of total ESIs up to that observed in the placebo group . There was some evidence that infections caused by Gram-negative bacteria had less severe consequences than staphylococcal infections . It is concluded that application of nasal mupirocin to nasal carriers of Staphylococcus aureus may be cost-saving in patients undergoing cardiac surgery or haemodialysis but, if the analysis is restricted to the cost of management of ESIs, it may not be cost-saving in CAPD . However, reducing the risk of staphylococcal ESI may reduce the risk of catheter loss and subsequent transfer to haemodialysis and this merits further study. J Hosp Infect, 1998 Sep, 40 Suppl B, S25 - 9 Reduction of surgical site infections in major surgery by elimination of nasal carriage of Staphylococcus aureus; Kluytmans J; Staphylococcus aureus has long been recognized as an important pathogen in human disease . Staphylococcal infections occur regularly in hospital patients and, despite antibiotic therapy, have severe consequences . An increasing number of such infections are caused by methicillin-resistant S . aureus (MRSA) strains, many of which have become multi-resistant to treatment . In an unblinded intervention trial, with historical controls, perioperative nasal carriage of S . aureus was eliminated using mupirocin nasal ointment . A significant reduction in surgical site infection was observed post-intervention in the treated group of patients . No resistant to mupirocin was observed . The results of this study warrant a prospective randomized, placebo-controlled study to confirm the efficacy of mupirocin. J Hosp Infect, 1998 Sep, 40 Suppl B, S13 - 23 Reduction of Staphylococcus aureus nasal carriage and infection in dialysis patients; Herwaldt LA; Numerous studies conducted in different countries and in different populations of patients on dialysis have consistently documented that a large proportion of such patients carry Staphylococcus aureus in their nares and that the risk of them becoming infected with their own strains is quite high . Furthermore, S . aureus infections can cause considerable morbidity and mortality in these patients . Thus, decolonization of the nares may prevent S . aureus infections and the attendant complications . The published data that support the use of rifampicin, intranasal mupirocin and povidone-iodine to prevent S . aureus infections in patients on dialysis are reviewed in detail. J Hosp Infect, 1998 Sep, 40 Suppl B, S3 - 11 The nose: an underestimated source of Staphylococcus aureus causing wound infection; Casewell MW; For the last fifty years, the nose has been intermittently recognized and targeted as a source of Staphylococcus aureus causing surgical site infection . In London in 1959, Williams and co-workers established for the first time that nasal carriers had increased rates of surgical sepsis compared with non-carriers . For half of these patients, the source was the patient's own nose . Post-admission acquisition of tetracycline-resistant strains was associated with even higher rates of infection . The increasing appearance of epidemic methicillin-resistant S . aureus (MRSA) in the 1980s rekindled interest in these (largely overlooked) studies, when the elimination of nasal carriage by topical mupirocin proved pivotal for the control of MRSA in Northern Europe and elsewhere . In the late 1980s and 1990s, Boelaert, Holton and others, appreciating the work performed forty years previously, used nasal mupirocin for the successful prevention of sepsis with S . aureus in patients on haemodialysis and continuous ambulatory peritoneal dialysis without incurring problems with mupirocin resistance . In 1995, Kluytmans and colleagues demonstrated that nasal carriage of S . aureus is a significant risk factor for wound infection after cardiac surgery . Towards the year 2000, the use of prophylactic nasal mupirocin for the prevention of serious sepsis in major clean surgery is emerging as a plausible and exciting new strategy. J Hosp Infect, 1998 Sep, 40(1), 61 - 5 Investigation of gaseous ozone for MRSA decontamination of hospital side-rooms; Berrington AW et al.; A domestic, gaseous ozone generator was investigated for use in the decontamination of hospital side-rooms that have housed patients colonized with methicillin-resistant Staphylococcus aureus (MRSA) . Three models of bacterial contamination were used . These were exposed to ozone generation in a standard hospital side-room for 4 and 7 h . A methicillin-sensitive and a methicillin-resistant strain of S . aureus were compared . Ozone concentrations of 0.14 ppm were reached, levels which are sufficient to cause mild pulmonary toxicity . Bacterial counts were reduced in the vicinity of the gas generator in most instances, but the effect elsewhere in the room was, at best, limited . MRSA appeared more resistant to the effects of ozone than methicillin-sensitive S . aureus . We conclude that the device tested would be inadequate for the decontamination of such hospital side-rooms. Ann Acad Med Singapore, 1998 May, 27(3), 358 - 62 Use of central venous lines in paediatrics--a local experience; Chua MC et al.; Central venous catheters are widely used in the care of critically ill patients . This paper reviews our experience with central lines in paediatric patients requiring intensive care, between the period August 1994 and August 1995 . A total of 57 insertions were performed in 40 patients, all less than 12 years of age . We found that the most common indication for catheter use was nutritional support (40%) . The overall complication rate was 58% . Catheter-related infection was the most serious problem, occurring in 32% of all insertions . Coagulase-negative Staphylococcus aureus was the organism most frequently isolated . Maintenance problems affected 17 of our catheters in which 9 were blocked . Both infected and blocked catheters were promptly removed . We had 3 cases of perforation and 2 cases of thrombosis . There were no deaths directly attributed to catheter use . Recommendations made include: 1) staff education and new guidelines for catheter care, 2) use of bacteria filters, 3) careful prospective monitoring of catheter infection rate, 4) heparinisation when infusion rate less than 2 ml/h, 5) eliminate use of stiff polyethylene catheters and 6) routine confirmatory X-ray or waveform monitoring before catheter use, if possible . We concluded that central venous catheterisations greatly facilitated the management of our patients . However, one must bear in mind that the use of such catheters is associated with problems which must be recognised early and promptly treated and, if possible, prevented with safe practice. Wound Repair Regen, 1998 Mar-Apr, 6(2), 149 - 56 Nonviable Staphylococcus aureus and its peptidoglycan stimulate macrophage recruitment, angiogenesis, fibroplasia, and collagen accumulation in wounded rats; Kilcullen JK et al.; We have previously shown that local application at the time of operation of Staphylococcus aureus, nonviable S . aureus, its cell wall, or S . aureus peptidoglycan accelerates wound healing . We hypothesized that this effect is due to both direct and indirect mechanisms, among which is an increase in the inflammatory response to wounding, resulting in an increase in macrophages, angiogenesis, and fibroblasts . Twenty-seven Sprague-Dawley male rats were anesthetized, and two 7-cm paravertebral skin incisions were made . Four polyvinyl alcohol sponges, two on each side, containing either 100 microliter of isotonic saline or 0.5 mg of nonviable S . aureus or S . aureus peptidoglycan in 100-microliter saline were implanted subcutaneously . Nonviable S . aureus or S . aureus peptidoglycan (860 microgram/cm incision) in 200-microliter saline were inoculated into the incisions at closure . The rats ate a commercial rat chow and drank tap water ad libitum throughout . After days 3 and 7 postwounding, rats were euthanized, and tissues were examined for immunohistochemical features of reparative tissue using ED-1, Factor VIII, and vimentin antibodies, markers for monocyte/macrophages, endothelial cells, and mesenchymal cells (including fibroblasts), respectively . Incisions treated with nonviable S . aureus or S . aureus peptidoglycan showed more macrophages along and deep in the wound tract 7 days postoperatively . Nonviable S . aureus or S . aureus peptidoglycan-treated sponges were surrounded and penetrated by much larger capsules of reparative tissue than saline-treated sponges at both 3 and 7 days . Neutrophil influx was much greater in nonviable S . aureus or S . aureus peptidoglycan-treated sponges, especially in central regions, and there were many more ED-1-stained macrophages in distinct geographic locations, specifically, the more peripheral-cortical areas . Some clustering of macrophages occurred around areas of invasion by reparative tissue into the surrounding subcutaneous fat and within the interstices of the sponges at the interface between reparative tissue and acute inflammatory cells . In contrast, saline-treated sponge reparative tissue had significantly fewer macrophages, much thinner and flimsy reparative tissue, with proportionately fewer macrophages clustering centrally . There were many more mesenchymal cells (notably fibroblasts) and new blood vessels and much more reparative collagen in the nonviable S . aureus or S . aureus peptidoglycan-treated sponges . We conclude that local application of nonviable S . aureus or S . aureus peptidoglycan at wounding induces an increased number and alteration in location of macrophages, increased influx (or proliferation) of mesenchymal cells (notably fibroblasts), and increased angiogenesis and reparative collagen accumulation, as well as increasing the overall acute inflammatory response to wounding. Rev Bras Enferm, 1997 Apr-Jun, 50(2), 291 - 6 {Nursing research on the use of venous catheters for hospitalized patients}; Utyama IK et al.; This report shows that intravenous catheterisation has been required for 59.4% of the patients in this sample . The main reason identified for catheterisation was the difficulty in accessing peripheral veins . The wound dressing was applied for 100% of the patients and the procedure itself was carried out each 48 hours in 89.8% of them . The intravenous catheter stood for 10 days in 52% of the cases and it was removed in 34.8% for intravenous drugs interruption and the other reasons were related as to technical problems . The most present microorganism was Staphylococcus aureus in 20.3% of the catheter cultures. Rev Med Interne, 1998 Mar, 19(3), 185 - 91 {Rare hyperimmunoglobulinemia E syndromes}; Katchourine I et al.; Total immunoglobulin E levels are very frequently measured, particularly for allergy . Their high levels are ordinarily found in atopy and parasitosis . This article subject is to determine rare diseases where high levels of total immunoglobulin E are found except in classic diseases . These rare hyper-immunoglobulin E syndromes are more often associated with immunologic deficiency more or less intense that lead to viral, fungal and bacterial infections . Buckley syndrome: deep infections due to Staphylococcus aureus, chronic dermatitis . Netherton disease: cutaneous and hair disease, allergic manifestations . Wiskott-Aldrich syndrome: thrombocytopeny and -pathy, dermatitis . Di-George disease: hypoparathyroidism, infections . Selective deficit of immunoglobulin A: recurrent infections . It is difficult to differentiate between atopic dermatitis and Buckley syndrome and Netherton disease because of eczematiform chronic dermatitis and high-levels of total immunoglobulin E found in all these diseases. Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1995 Feb, 28(1), 47 - 58 {Coagulase type and antimicrobial susceptibility of Staphylococcus aureus isolated from various areas in Taiwan}; Hsieh SE et al.; A total of 129 strains of Staphylococcus aureus, isolated from clinical specimens in Taiwan between February 1992 and December 1993, were subjected to coagulase typing and susceptibility testing to 21 kinds of antimicrobial agents using Pasco MIC Gram-positive panels . In the determination of minimum inhibition concentration (MIC), there were 94 strains (72.9%) resistant to penicillin and ampicillin, 54 strains (41.9%) resistant to tetracycline and erythromycin, and 21 strains (16.3%) resistant to oxacillin (oxacillin-resistant S . aureus; ORSA), but none of them was resistant to vancomycin or nitrofurantoin . As the susceptibility of the isolates from four different geographic districts was compared, no statistical difference was found except that the resistance rate to penicillin and ampicillin was higher in southern Taiwan, and resistance rate to rifampin and gentamicin was higher in central Taiwan . The ORSA strains were all resistant to penicillin, ampicillin, tetracycline; 95.2% of the strains were resistant to gentamicin, tobramycin and erythromycin . The resistance rates to drugs tested for ORSA strains were statistically higher than those for OSSA strains except vancomycin, nitrofurantoin, trimethoprim/sulfamethoxazole and ampicillin/sulbactam . In the coagulase typing of 127 strains, Type IV, III and VII were most frequently encountered . Among the coagulase types, Type IV was mostly encountered in the North, the South and the East of Taiwan; Type III was mostly encountered in central Taiwan . Among the ORSA strains, coagulase Type III was most predominant (85%) . In conclusion, analysis of an antibiogram is easy to perform and the results can provide clinicians not only with correct guides for patient treatment but also with a useful tool for epidemiological studies . However, if antibiogram and coagulase typing are carried out simultaneously, results will be more reliable in epidemiological studies, including nosocomial infection survey. Tidsskr Nor Laegeforen . 1998 Sep 10;118(21):3282. {Tropical pyomyositis}; Iversen PO et al.; Tropical pyomyositis is rarely observed among permanent residents of temperate or cold climates and is, to our knowledge, not described among Norwegians . It is a clinical entity comprising general symptoms of infection and abscesses in skeletal muscles . We present one case of tropical pyomyositis acquired in the Dominican Republic . The patient, a female, had an insidious progression of the disease with fever, chills, and general malaise . On admission she had also developed multiple abscesses affecting muscles of the extremities . She required surgical drainage in addition to antibiotics . Cultures from purulent material revealed Staphylococcus aureus. Clin Infect Dis, 1998 Sep, 27(3), 478 - 86 Outcome of Staphylococcus aureus bacteremia according to compliance with recommendations of infectious diseases specialists: experience with 244 patients; Fowler VG Jr et al.; To determine whether recommendations of infectious diseases specialists affect outcome for patients, we evaluated 244 hospitalized patients with Staphylococcus aureus bacteremia . We offered our management recommendations to each patient's physicians and then assessed the clinical outcome for both patients for whom our consultative advice was followed and those for whom our advice was not heeded . All patients were followed up for 12 weeks after their first positive blood culture . Our management advice was followed for 112 patients (45.9%) and partially or completely ignored for 132 patients (54.1%) . Patients for whom our recommendations were followed were more likely to be cured of their S . aureus infection and less likely to relapse (P < .01), despite having significantly more metastatic infections (P < .01) at the outset of therapy, than were those for whom our recommendations were not followed . Failure to follow recommendations to remove an infected intravascular device was the most important risk for treatment failure . After controlling for other factors, logistic regression analysis revealed that patients whose intravascular device was not removed were 6.5 times more likely to relapse or die of their infection than were those whose device was removed . Our findings suggest that patient-specific management advice by infectious diseases consultants can improve the clinical outcome for patients with S . aureus bacteremia. FEMS Microbiol Lett, 1998 Sep 15, 166(2), 225 - 30 A human transferrin-binding protein of Staphylococcus aureus is immunogenic in vivo and has an epitope in common with human transferrin receptor; Lim Y et al.; To understand human immune responses against the human transferrin-binding protein of Staphylococcus aureus (SA-tbp), we examined cell wall proteins from S . aureus ATCC 6538 using human convalescent sera, and a monoclonal antibody specific for human transferrin receptor (McAb-HTR) . The SA-tbp, detected by immunoblot assay, was iron-repressible, reacted with the convalescent sera, and cross-reacted with McAb-HTR . Immunoelectron microscopy probed with McAb-HTR showed a reaction zone around the test strain from the deferrated BHI . After being preincubated with an S . aureus-bacteremic serum, the electroblot of the SA-tbp still reacted with McAb-HTR, but not with human transferrin-horseradish peroxidase conjugate . We conclude, there are at least two kinds of epitopes in the SA-tbp; one able to bind to human transferrin is immunogenic in humans, but the other sharing epitopes common with human transferrin receptor is not immunogenic in humans. Pathology, 1998 Aug, 30(3), 299 - 303 Heterogeneous expression of fusidic acid resistance in Staphylococcus aureus with plasmid or chromosomally encoded fusidic acid resistance genes; O'Brien FG et al.; Fusidic acid resistance expression in a methicillin susceptible Staphylococcus aureus strain (WBG1576), which carries fusidic acid resistance on plasmid pUB101, and a prevalent Western Australian methicillin-fusidic acid resistant strain (WBG8287) were compared . WBG8287 carries fusidic acid resistance on the chromosome and its plasmid content has no effect on the levels of this resistance . WBG1576 and WBG8287 exhibited similar heterogeneous populations in respect to fusidic acid resistance levels in population analyses . A high-level fusidic acid resistant mutant of WBG1576 (BE8) had alterations in Smal chromosomal profiles, but not in plasmid size or resistance expression . Mutations causing increased fusidic acid resistance in WBG1576 are chromosomally located . A high-level fusidic acid resistant mutant of WBG8287 (BE3) had no alterations in Smal chromosomal profiles, or plasmid content and resistances . Comparison of resistance levels to kanamycin and spectinomycin, between high-level resistant colonies of WBG8287 and WBG8287, indicate that mutations in the chromosomal gene fusA, which encodes elongation factor-G, are probably the cause of the increased resistance levels observed in these mutant strains. Clin Infect Dis, 1998 Sep, 27(3), 543 - 50 Selective screening of carriers for control of methicillin-resistant Staphylococcus aureus (MRSA) in high-risk hospital areas with a high level of endemic MRSA; Girou E et al.; Screening for methicillin-resistant Staphylococcus aureus (MRSA) carriage in patients at risk was evaluated as part of a control program in a 26-bed medical intensive care unit (ICU) of a university hospital with a high level of endemic MRSA . Control measures included isolation and barrier precautions, skin decolonization with chlorhexidine of patients from whom MRSA was recovered, and mupirocin treatment of nasal carriers of MRSA . Of 3,686 patients admitted during a 4-year period, 44% were screened, which occurred during admission for 38%; MRSA was recovered from 293 patients (8%) . There were 150 imported cases and 143 ICU-acquired cases, of which 51% and 45%, respectively, were first identified through screening . Nasal swab cultures identified 84% of MRSA carriers . The incidence of all ICU-acquired cases and of acquired colonization or infection decreased from 5.8% and 5.6% to 2.6% and 1.4% (P = .002 and P < .001), respectively, whereas that of imported cases remained unchanged (range, 3.8% to 4.3%; P = .8) . Selective screening for nasal carriage during admission to high-risk areas may contribute to identification of a substantial proportion of cases of MRSA and to early implementation of effective control measures. Clin Infect Dis, 1998 Sep, 27(3), 458 - 62 Is bacterial tracheitis changing? A 14-month experience in a pediatric intensive care unit; Bernstein T et al.; Bacterial tracheitis is characterized by acute upper-airway obstruction and purulent secretions within the trachea . Historically, affected children were young, stridorous, and toxic-appearing and required tracheal intubation, and morbidity and mortality were significant . Staphylococcus aureus was the most common organism involved . During the 14 months of this retrospective study, 46 children were admitted to the pediatric intensive care unit because of this diagnosis, and their medical records were reviewed . Compared with those in previous reports, children in this study were older (mean +/- standard error of the mean {SEM}, 69.3 +/- 6.8 months) and less toxic . Only 26 (57%) of 46 patients required tracheal intubation . Intubated patients were significantly younger than nonintubated patients (mean +/- SEM, 46.9 +/- 6.5 vs . 98.9 +/- 9.9 months) . Moraxella catarrhalis was identified in 12 (27%) of 45 bacterial respiratory cultures, while influenza A virus was recovered from 18 (72%) of 25 viral respiratory cultures . There were no major complications . This series represents the largest reported cohort of patients with this condition and suggests an epidemiological change toward a less morbid condition. Pathol Biol (Paris), 1998 Apr, 46(4), 227 - 34 {Methicillin-resistant Staphylococcus aureus: evolution and epidemiology, clinical impact, and prevention}; Brun-Buisson C; Methicillin-resistant Staphylococcus aureus (MRSA) strains have become endemic in hospitals in many countries . In most cases, epidemic clones emerge occasionally on the endemic background . Early institution of simple measures aimed at preventing hand-borne cross-contamination has resulted in low rates of MRSAs in a few countries of northern Europe . Although environmental contamination plays a role in some cases, colonized or infected patients are the main reservoir . Three significant problems have been identified in countries with high levels of endemicity, namely identification of the reservoir in the hospital, including detection of healthy carriers who are readmitted; notification of departments that receive healthy carriers transferred from another department in the same hospital or from another hospital; and the increasing and hard-to-control reservoir of resistant bacteria in extended-care facilities . These three problems are related to the current level of endemicity and to the risk of dissemination of resistant strains during hospital-to-hospital patient transfers . The technical responses to these problems vary with the environment and available means . They include detection of carriers in high-risk units, use of standardized reporting forms for patient transfers, and recording of the presence of MRSA in computerized medical records . Decontamination of carriers can help to prevent cross-contamination in some situations . It is of the utmost importance that all the facets of MRSA endemicity be taken into account at the level of the hospital as a whole and that all decision makers and health care providers make a commitment to reducing MRSA endemicity. Pathol Biol (Paris), 1998 Apr, 46(4), 213 - 6 {Antibiotic resistance in the European countries}; Carlet J; The overall level of resistance to antimicrobials is high in Europe, most notably in intensive care units . Variations exist across countries, with the levels being highest in southern Europe . Staphylococcus aureus remains the most hazardous pathogen . Glycopeptide-resistant enterococci are relatively infrequent in Europe for the time being . Differences in measures used to control antimicrobial use and to prevent cross-contamination probably explain country-to-country differences in resistance levels . Substantial efforts are needed in both these areas in many countries, including France. Pathol Biol (Paris), 1998 Jun, 46(6), 435 - 41 {Pore-forming leukotoxins from Staphylococcus aureus: variability of the target cells and 2 pharmacological processes}; Prevost G et al.; The staphylococcal bi-component leukotoxins constitute a family included in the super-family of the beta-sheet-structured pore-forming toxins . They may be produced by Staphylococcus aureus and by Staphylococcus intermedius and their target cells vary according to the molecules . The mode of action proceeds by the sequential binding of the class S proteins, then by that of the class F proteins at the surface of the membranes . Then, the activation of cellular calcium-channels precedes the pore formation which seems to be sensitive to several monovalent cations . The cell response is inflammatory and includes the neosynthesis as well as the secretion of leukotriene B4, interleukin -8, histamine . The injection of leukotoxins to rabbits generates cell chemotaxis , vasodilatation, and tissue necrosis . The association of the production of leukotoxins with clinical syndromes concerns several aspects of the pathology of S . aureus, and confers to these leukotoxins an important role of virulence factors. J Allergy Clin Immunol, 1998 Sep, 102(3), 428 - 35 Reduced prevalence of allergic disease in patients with multiple sclerosis is associated with enhanced IL-12 production; Tang L et al.; BACKGROUND: We previously showed that the prevalence of allergic disease is decreased in patients with multiple sclerosis (MS); however, the mechanisms that explain this finding have not previously been defined . OBJECTIVES: We have demonstrated that protection of patients with MS from allergic disease may be caused by the production in monocytes from these patients of elevated quantities of IL-12 compared with that observed in monocytes from individuals with allergies . METHODS: Purified monocytes from peripheral blood of subjects with or without allergies and from individuals with MS were directly stimulated with Staphylococcus aureus Cowan strain I in the absence of T cells . IL-12 was quantitated by a sensitive reverse transcription, competitive PCR . RESULTS: IL-12 production was 5-fold greater in monocytes from patients with MS (n = 11) than that from individuals with allergies (n = 10) (for subjects with MS, 1.90+/-0.18 vs 1.24+/-0.19 log10 fmol/microL for individuals with allergies) (P = .02) . Although the production of IL-12 in monocytes from patients with MS was slightly higher than that from subjects without allergies, this difference was not statistically significant . CONCLUSIONS: IL-12 production in individuals with MS is much greater than in individuals with allergies . Because IL-12 induces TH1 cytokine synthesis and reduces the production of TH2 cytokines, which amplify and prolong allergic inflammation, these studies suggest that enhanced IL-12 production may protect individuals with MS from the development of allergy but may predispose such individuals toward autoimmune inflammation in the central nervous system. Ann Chir Plast Esthet, 1997 Feb, 42(1), 70 - 4 {Covering of a thoraco-lumbar defect by omentoplasty}; Le Fourn B et al.; With a case of thoraco-lumbar defect, the authors discuss about different procedures to cover it . In this place, the better procedure is certainly the latissimus dorsi flap, in all combinations . The indication for omentoplasty at this spinal site should not be performed by first intention but by exclusion of other procedures, as in the case considered by the authors . It was a 37-year-old man, paraplegic from the age of 16, with a deep chronic spinal wound, secondary to sepsis of a posterior segmental fixations . A staphylococcus aureus infection which developed as a surgical complication was initially treated with antibiotics and surgical cleaning procedures without removing instrumentation . However, the infection remained active and the material was finally removed . Spinal immobilisation was strengthened by external fixation . The area was cleared of all suspect material, including bone graft, leaving a wide back-wound open to the spine . Spontaneous healing was first attempted, but the size and the chronicity of the wound led us to use pedicled greater omentum to close the defect . The omentum was pedicled on the right gastroepiploic vessels and transferred to the back wound through the posterior abdominal wall muscles, next to the right kidney . This procedure allows rapid healing . In association with suitable antibiotics, it has prevented any recurrent infection after 18 months of follow-up . It was no feasible to cover the wound with a latissimus dorsi flap, considering the importance of this muscle in the movements of a paraplegic and considering the initial impossibility of removing the external fixation. Br J Dermatol, 1998 Aug, 139(2), 319 - 24 Chronic staphylococcal scalded skin syndrome; Shelley ED et al.; Staphylococcal scalded skin syndrome (SSSS), not previously recorded as a chronic disease, persisted for 2 years in a 50-year-old woman with epilepsy and cerebellar ataxia . Lesions initially suggestive of erythema multiforme and toxic epidermal necrolysis evolved over 2 years into those typical for SSSS, with extensive erosions and subcorneal blisters, showing an epidermal split at the granular cell layer . Exfoliatin A-producing phage I-III Staphylococcus aureus, previously linked only to acute mild adult cases of SSSS, was cultured from purulent discharge in the patient's eyes, ears and open skin lesions . The roles of epilepsy and antiepileptic medications are discussed as possible predisposing factors. J Antibiot (Tokyo), 1998 Aug, 51(8), 750 - 6 Antibiotic activities and affinities for bacterial cell wall analogue of N-demethylvancomycin and its derivatives; Yan H et al.; N-Demethylvancomycin, which has been clinically used in China, is one member of vancomycin group (glycopeptide) antibiotics . It differs from vancomycin only in that methyl group on the amino group of the N-terminal residue of vancomycin has been replaced by H . By reductive alkylation of N-demethylvancomycin, we synthesized N-alkyl and N,N'-dialkyl N-demethylvancomycins, which closely correlated with vancomycin in structure . The association constants of the complexes of N-demethylvancomycin and its analogues with di-N-Ac-L-Lys-D-Ala-D-Ala and the antibiotic activity against Staphylococcus aureus of the glycopeptides were determined . Results showed that N-demethylvancomycin has higher affinity for bacterial cell wall analogue di-N-Ac-L-Lys-D-Ala-D-Ala and more potent antibiotic activity against Staphylococcus aureus than vancomycin . Both N-alkylation and N,N'-dialkylation of N-demethylvancomycin reduced the affinity and antibiotic activity . The longer the alkyl groups, the less potent antibiotic activities and lower affinities have the glycopeptides . The antibiotic activities against Staphylococcus aureus of N-demethylvancomycin and its analogues roughly parallel their affinities for di-N-Ac-L-Lys-D-Ala-D-Ala. Res Microbiol, 1998 Jul-Aug, 149(7), 497 - 507 Genetic diversification of methicillin-resistant Staphylococcus aureus as a function of prolonged geographic dissemination and as measured by binary typing and other genotyping methods; van Leeuwen W et al.; The aim of the present study was to determine the extent of genome evolution among methicillin-resistant Staghylococcus aureus (MRSA) strains . Three different collections of strains were analysed, comprising locally, nationally and internationally disseminated genotypes . Various genotyping assays displaying different levels of resolution were used . Geographically and temporally diverse MRSA strains comprised the international group . MRSA strains recovered during an outbreak in a New York City hospital and Portuguese MRSA isolates, all resembling the so-called Iberian clone, were included in the local and national collections, respectively . Genotypes were determined by genome scanning typing techniques and procedures which analyse specific DNA elements only . The outbreak strains showed subclonal variation, whereas the Portuguese isolates displayed an increased number of genotypes . Among the epidemiologically unrelated MRSA strains, the different genotyping techniques revealed a wide heterogeneity of types . Different typing techniques appeared to show different levels of resolution, which could be correlated with the extent of geographic spread; the more pronounced the spread, the higher the degree of genome evolution . Binary typing and randomly amplified polymorphic DNA analysis are the typing methods of choice for determining (non)identity among strains that have a recent common ancestor and have undergone yet limited dissemination. Ann Acad Med Stetin, 1998, Suppl 41, 1 - 72 {Aortoiliac graft infection as a diagnostic and treatment problem}; Gutowski P; Aortoiliac graft infection occurs in 2-6% of patients with such prosthesis . This condition is seldom properly diagnosed by conventional radiographic methods, leading to high morbidity and mortality . Clinically, the diagnosis of aortic graft infection is difficult because patients may have a variety of nondescript clinical complaints . The diagnosis of graft infection when associated with minimal or absent clinical signs of low-grade infection is uncertain, but is critically important to avoid frequently catastrophic complications such as sepsis, gastrointestinal hemorrhage, and suture line disruption . Aim of the study: identification of bacterial flora present in aortoiliac graft infection; the presentation of my own experience in the detection of aortoiliac graft infection with special description of isotopic study with WBC labeled 99mTc HM-PAO and estimation of the usefulness of this test in comparison with computed tomography, ultrasonography, fistulography and angiography; evaluation of the usefulness of various treatment methods; establishing principles (algorithm) of diagnostic and therapeutic procedures in the case of the suspicion of aortoiliac graft infection . As many as 1190 patients with implanted aortoiliac graft in 1986-1996 at the General and Vascular Surgery Clinic in Szczecin were studied (Tab . 2) . Thirty-one patients in the study had deep aortoiliac graft infection (Tab . 3), while 9 patients had, in addition, prosthetic-enteric fistulae and 1 had arterio-enteric secondary fistulae . The group of 31 patients with deep graft infection, that is 2.6% of all patients (1190), had aortoiliac graft implanted at the mentioned time period (Tab . 4, 5) . Test results for detection of graft infection have been analysed (Tab . 17) . The results of isotopic investigation (Tab . 16), computed tomography (Tab . 11), ultrasonography (Tab . 13), fistulography (Tab . 14) and angiography (Tab . 15) were compared with intraoperative state or in a case of exclusion of infection, with results of follow up . Results of various paths of treatment were estimated (Tab . 18) . Based on performed cultures most common bacterial flora from infected grafts, was identified (Tab . 9, 10) . The sensitivity of the isotopic study with labeled white blood cells in detection of graft infection was 88%, specificity was 97%, accuracy 93%, positive predictive value 96% . Other useful diagnostic procedures in detection of aortic graft infection are: computed tomography with an accuracy of 75%, endoscopic investigation useful in detection of arterio-enteric fistulae with an accuracy of 50% and ultrasonography with an accuracy of 35.5% (Tab . 17) . The choice of the best treatment is still controversial . In my material total excision of infected graft and extraanatomic revascularization were burdened with 50% mortality rate . Among patients treated less radically the mortality rate was considerably lower . In a group of patients with the excision of the infected graft only, the mortality rate was 9% but the amputation rate was 36.4% and in a group of patients with excision of infected graft and reconstruction in situ, the mortality rate was 25% (Tab . 18) . Taking into consideration our results, less aggressive methods of aortic graft infection treatment such as the excision of the infected part of the prosthesis with or without in situ revascularization if only possible should be recommended . Most common in bacterial cultures from infected aortoiliac grafts with prosthetic-enteric fistulae were Escherichia coli found (Tab . 10) . In infections without fistula various types of Staphylococcus aureus were identified (Tab . 9) . CONCLUSIONS: 1 . In cases of aortoiliac graft infection the most common cultured bacteria are found to be Staphylococcus aureus . If there is additional prosthetic-enteric fistula Escherichia coli is the most common cultured bacteria . 2 . In a case there was suspicion of aortoiliac graft infection, proper diagnostic procedures are most important for effective ma J Pharmacol Exp Ther, 1998 Oct, 287(1), 128 - 36 Epithelial ion transport and barrier abnormalities evoked by superantigen-activated immune cells are inhibited by interleukin-10 but not interleukin-4; Lu J et al.; Many studies have indicated an association between bacteria and the severity of enteric secretory or inflammatory disorders . We previously showed that monolayers of human T84 epithelial cells display altered ion transport and permeability after coculture with Staphylococcus aureus enterotoxin B (SEB, a model superantigen)-activated immune cells, where interferon-gamma and tumor necrosis factor-alpha were key mediators in the pathophysiology . Here we examined whether the regulatory Th2-type cytokines, interleukin (IL)-10 and IL-4, could prevent these epithelial irregularities . T84 monolayers were cocultured with human peripheral blood mononuclear cells (PBMC) or T cell-enriched, monocyte-depleted PBMC (T + B cells) +/- SEB for 20 hr in the presence or absence of IL-10 or IL-4 . Subsequently, T84 monolayers were mounted in Ussing chambers and ion transport (short-circuit current (Isc) and DeltaIsc evoked by forskolin) and permeability (ion resistance and probe fluxes) were assessed . IL-10 dose-dependently inhibited the increased T84 permeability and the reduced responsiveness to forskolin that were evoked by coculture with SEB-activated PBMC or T + B cells . Similar changes in T84 function occurred in response to conditioned medium from SEB-activated immune cells; however, addition of IL-10 to the conditioned medium did not prevent the changes in epithelial function . In contrast, when PBMC were stimulated with SEB in the presence of IL-10, the subsequent conditioned medium was less effective in evoking altered epithelial function . These data suggest that the affect of IL-10 was due to effects on the immune cells and not directly on the epithelium . In contrast to IL-10, IL-4 did not ameliorate any of the immune-mediated changes in T84 function . We conclude that IL-10 can reduce the epithelial functional changes caused by SEB-activated immune cells and this data adds further support for IL-10 immunotherapy in the treatment of intestinal secretory or inflammatory disorders. Acta Crystallogr D Biol Crystallogr, 1998 Mar 1, 54 ( Pt 2), 276 - 8 Crystallization of the alpha-hemolysin heptamer solubilized in decyldimethyl- and decyldiethylphosphine oxide; Song L et al.; Crystals of the alpha-hemolysin heptamer, a transmembrane pore-forming toxin from Staphylococcus aureus, have been grown using the nonionic detergents n-decyldimethyl- and n-decyldiethylphosphine oxide, phosphorus homologs of the ionic amine oxide detergents . Five crystal forms were obtained, one of which diffracted X-rays to 3 A resolution . This crystal form displayed elements of pseudo mm symmetry in screened precession photographs yet it was triclinic with unit-cell dimensions a = 173, b = 173, c = 102 A, alpha = 92.6, beta = 94.8, gamma = 90.2 degrees. J Bone Joint Surg Am, 1998 Sep, 80(9), 1336 - 40 Anti-infective efficacy of antiseptic-coated intramedullary nails; Darouiche RO et al.; The coating of medical devices with antimicrobial agents has recently emerged as a potentially effective method for the prevention of device-related infections . We examined the anti-infective efficacy of intramedullary nails coated with an antiseptic combination of chlorhexidine and chloroxylenol in a rabbit model of device-related infection after fixation of an open tibial fracture . The rabbits were randomized to receive 2.8-by-100-millimeter stainless-steel tibial intramedullary nails that either were uncoated or were coated with antiseptic . After administration of anesthesia and preoperative antibiotic prophylaxis, a tibial fracture was created and then reduced with insertion of the intramedullary nail . A bacterial inoculum of 10(6) colony-forming units of Staphylococcus aureus was injected into the intramedullary canal, and the wound was sutured . Radiographs of the tibiae were made postoperatively, and the rabbits were monitored daily . They were killed at six weeks, or earlier if there was dehiscence of the wound, the fracture became grossly unstable, or the rabbit failed to thrive . The use of the antiseptic-coated nails was associated with a significantly lower rate of device-related osteomyelitis (two of twenty-two; 9 per cent) than the use of the uncoated nails (thirteen of twenty-one; 62 per cent) (p = 0.0003) . The radiographic and histopathological findings were generally similar in the two groups of rabbits . Antiseptic agents were not detected in serum . The results suggest that antiseptic-coated fracture-fixation devices provide significant local protection against Staphylococcus aureus, which is the most common cause of infections related to orthopaedic devices. Infect Control Hosp Epidemiol, 1998 Aug, 19(8), 552 - 9 Methicillin-resistant Staphylococcus aureus and its relationship to antimicrobial use: possible implications for control; Monnet DL; The control of methicillin-resistant Staphylococcus aureus (MRSA) is still an unresolved issue in numerous healthcare institutions worldwide . Guidelines for the control of MRSA in hospitals focus on measures to control cross-transmission and prevent colonization, but rarely specifically mention the control of antimicrobial use . We reviewed the different types of evidence for a causal relationship between MRSA and antimicrobial use by classifying them in four categories: consistent associations, dose-effect relationships, concomitant variations, and arguments to support a plausible biological model to explain this relationship . Although the relative participation of cross-transmission and antimicrobial selection pressure in the level of MRSA observed in a healthcare setting remains to be determined, we found lines of evidence to support the existence of a relationship between MRSA and antimicrobial use in each of the four categories . This review points out the relative lack of studies specifically designed to investigate this aspect of MRSA epidemiology and the need to implement such studies quickly . In the meantime, the results presented here should encourage the implementation of antimicrobial-use improvement programs in hospitals in addition to existing infection control measures, which are still a priority in countries with high MRSA prevalence. Gene, 1998 Sep 28, 219(1-2), 9 - 17 Sequence of the putative alanine racemase operon in Staphylococcus aureus: insertional interruption of this operon reduces D-alanine substitution of lipoteichoic acid and autolysis; Kullik I et al.; A gene cluster comprising the alanine racemase gene alr was identified 5' to the sigB operon in Staphylococcus aureus . It is flanked upstream by four ORFs of which one shows similarity to the dpj gene of Escherichia coli, and downstream by two ORFs of which the last shows similarity to the E . coli pemK gene . Preliminary data suggest that the seven ORFs orf1-orf2-orf3-dpj-alr-orf6-pemK may form an operon . Disruption of the proposed operon by insertional mutagenesis leads to a drastic loss in the d-alanine (d-Ala) substitution of lipoteichoic acid and to delayed autolysis, without affecting the d-Ala substitution of the wall teichoic acid. Antimicrob Agents Chemother, 1998 Oct, 42(10), 2739 - 44 In vitro studies of pharmacodynamic properties of vancomycin against Staphylococcus aureus and Staphylococcus epidermidis; Lowdin E et al.; The bactericidal activities of vancomycin against two reference strains and two clinical isolates of Staphylococcus aureus and Staphylococcus epidermidis were studied with five different concentrations ranging from 2x to 64x the MIC . The decrease in the numbers of CFU at 24 h was at least 3 log10 CFU/ml for all strains . No concentration-dependent killing was observed . The postantibiotic effect (PAE) was determined by obtaining viable counts for two of the reference strains, and the viable counts varied markedly: 1.2 h for S . aureus and 6.0 h for S . epidermidis . The determinations of the PAE, the postantibiotic sub-MIC effect (PA SME), and the sub-MIC effect (SME) for all strains were done with BioScreen C, a computerized incubator for bacteria . The PA SMEs were longer than the SMEs for all strains tested . A newly developed in vitro kinetic model was used to expose the bacteria to continuously decreasing concentrations of vancomycin . A filter prevented the loss of bacteria during the experiments . One reference strain each of S . aureus and S . epidermidis and two clinical isolates of S . aureus were exposed to an initial concentration of 10x the MIC of vancomycin with two different half-lives (t1/2s): 1 or 5 h . The post-MIC effect (PME) was calculated as the difference in time for the bacteria to grow 1 log10 CFU/ml from the numbers of CFU obtained at the time when the MIC was reached and the corresponding time for an unexposed control culture . The difference in PME between the strains was not as pronounced as that for the PAE . Furthermore, the PME was shorter when a t1/2 of 5 h (approximate terminal t1/2 in humans) was used . The PMEs at t1/2s of 1 and 5 h were 6.5 and 3.6 h, respectively, for S . aureus . The corresponding figures for S . epidermidis were 10.3 and less than 6 h . The shorter PMEs achieved with a t1/2 of 5 h and the lack of concentration-dependent killing indicate that the time above the MIC is the parameter most important for the efficacy of vancomycin. Antimicrob Agents Chemother, 1998 Oct, 42(10), 2678 - 81 Inhibitory activities of gatifloxacin (AM-1155), a newly developed fluoroquinolone, against bacterial and mammalian type II topoisomerases; Takei M et al.; We determined the inhibitory activities of gatifloxacin against Staphylococcus aureus topoisomerase IV, Escherichia coli DNA gyrase, and HeLa cell topoisomerase II and compared them with those of several quinolones . The inhibitory activities of quinolones against these type II topoisomerases significantly correlated with their antibacterial activities or cytotoxicities (correlation coefficient {r} = 0.926 for S . aureus, r = 0.972 for E . coli, and r = 0.648 for HeLa cells) . Gatifloxacin possessed potent inhibitory activities against bacterial type II topoisomerases (50% inhibitory concentration {IC50} = 13.8 microg/ml for S . aureus topoisomerase IV; IC50 = 0.109 microg/ml for E . coli DNA gyrase) but the lowest activity against HeLa cell topoisomerase II (IC50 = 265 microg/ml) among the quinolones tested . There was also a significant correlation between the inhibitory activities of quinolones against S . aureus topoisomerase IV and those against E . coli DNA gyrase (r = 0.969) . However, the inhibitory activity against HeLa cell topoisomerase II did not correlate with that against either bacterial enzyme . The IC50 of gatifloxacin for HeLa cell topoisomerase II was 19 and was more than 2,400 times higher than that for S . aureus topoisomerase IV and that for E . coli DNA gyrase . These ratios were higher than those for other quinolones, indicating that gatifloxacin possesses a higher selectivity for bacterial type II topoisomerases. Antimicrob Agents Chemother, 1998 Oct, 42(10), 2590 - 4 Characterization of mutations in the rpoB gene that confer rifampin resistance in Staphylococcus aureus; Aubry-Damon H et al.; Mutations in the rifampin resistance-determining (Rif) regions of the rpoB gene of Staphylococcus aureus mutants obtained during therapy or in vitro were analyzed by gene amplification and sequencing . Each of the resistant clinical isolates, including five nonrelated clones and two strains isolated from the same patient, and of the 10 in vitro mutants had a single base pair change that resulted in an amino acid substitution in the beta subunit of RNA polymerase . Eight mutational changes at seven positions were found in cluster I of the central Rif region . Certain substitutions (His481/Tyr and Asp471/Tyr {S . aureus coordinates}) were present in several mutants . Substitutions Gln468/Arg, His481/Tyr, and Arg484/His, which conferred high-level rifampin resistance, were identical or in the same codon as those described in other bacterial genera, whereas Asp550/Gly has not been reported previously . Substitutions at codon 477 conferred high- or low-level resistance, depending on the nature of the new amino acid . The levels of resistance of in vivo and one-step in vitro mutants carrying identical mutations were similar, suggesting that no other resistance mechanism was present in the clinical isolates . On the basis of these data and the population distribution of more than 4,000 clinical S . aureus isolates, we propose </=0.5 and >/=8 microg/ml as new breakpoints for the clinical categorization of this species relative to rifampin. J Biochem (Tokyo), 1998 Oct, 124(4), 778 - 83 Pyrrolidone carboxyl peptidase from the hyperthermophilic Archaeon Pyrococcus furiosus: cloning and overexpression in Escherichia coli of the gene, and its application to protein sequence analysis; Tsunasawa S et al.; A gene for a pyrrolidone carboxyl peptidase (Pcp: EC 3.4.19.3, pyroglutamyl peptidase), which removes amino-terminal pyroglutamyl residues from peptides and proteins, has been cloned from the hyperthermophilic Archaeon Pyrococcus furiosus using its cosmid protein library, sequenced, and expressed in Escherichia coli . The DNA sequence encodes a protein containing 208 amino acid residues with methionine at the N-terminus . Analysis of the recombinant protein expressed in E . coli, including amino acid sequence analysis from the N-terminus by automated Edman degradation and ionspray mass spectrometric analysis of the peptides generated by enzymatic digestions with lysylendopeptidase and Staphylococcus aureus V8 protease, showed its primary structure to be completely identical with that deduced from its cDNA sequence . Comparison of the amino acid sequence of P . furiosus Pcp (P.f.Pcp) with those of bacterial Pcps revealed that a high degree of sequence identity (more than 40%) and conservation of the amino acid residues comprising the catalytic triad, Cys142, His166, and Glu79 . On the other hand, a unique short stretch sequence (positions around 175-185) that is absent in bacterial Pcps was found in P.f.Pcp . A similar stretch has also been reported recently in the amino acid sequence of Pcp from the hyperthermophilic Archaeon Thermococcus litoralis {Littlechild et al., in abstracts of the "International Congress on Exthermophiles '98" p . 58 (1998)} . To elucidate their contribution to the hyperthermostability of these enzymes, further structural studies are required. Int J Immunopharmacol, 1998 Jul, 20(7), 345 - 57 IL-6 functions in cynomolgus monkeys blocked by a humanized antibody to human IL-6 receptor; Imazeki I et al.; A humanized antibody to the human interleukin-6 receptor (IL-6R), hPM-1, blocked the interleukin-6 (IL-6) functions in normal cynomolgus monkey lymphocytes in vitro . The binding activity of hPM-1 to non-human primate IL-6R was examined in peripheral blood lymphocytes by flow cytometry . PM-1 recognized the IL-6R on T lymphocytes of cynomolgus and rhesus monkeys, but did not on those of marmosets . The homology between human IL-6R and its cynomolgus monkey counterpart was 97.3% in the extracellular domain of the amino acid sequence, as determined by DNA sequencing of the PCR product from peripheral blood mononuclear cells . PM-1 inhibited two functional parameters in vitro in cynomolgus monkeys: (1), T-cell proliferation stimulated by phytohemaglutinin and human IL-6; (2), Immunoglobulin G-production evoked by Staphylococcus aureus Cowan-1- and human IL-6-stimulated B lymphocytes . These data show that hPM-1 binds to and functionally blocks the cynomolgus monkey IL-6 receptors. Jpn J Antibiot, 1998 Jun, 51(6), 432 - 6 {Transferability of tazobactam/piperacillin (TAZ/PIPC) to cerebrospinal fluid of rabbit with meningitis caused by Staphylococcus aureus}; Haruta T et al.; The transferability of tazobactam/piperacillin (TAZ/PIPC) to cerebrospinal fluid (CSF) was studied employing rabbits with experimental meningitis caused by Staphylococcus aureus . 125 or 250 mg/kg of TAZ/PIPC was intravenously administered to rabbits with experimental meningitis then concentrations of TAZ and PIPC in CSF and serum were measured . In the group to which 125 mg/kg of TAZ/PIPC was administered, mean concentration of TAZ in CSF was 7.3 and 2.4 micrograms/ml at 30 and 60 min after administration, respectively, and concerning PIPC, it was 10.1 and 3.5 micrograms/ml, respectively . CSF/serum ratio of TAZ was 29.4% and 31.4%, respectively, and that of PIPC was 24.3 and 35.6%, respectively . In the group to which 250 mg/kg of TAZ/PIPC was administered, mean concentration of TAZ in CSF was 16.5 and 12.6% micrograms/ml, respectively, and concerning PIPC, it was 25.6 and 18.2 micrograms/ml, respectively . CSF/serum ratio of TAZ was 22.1 and 56.1%, respectively, and that of PIPC was 12.2 and 51.9%, respectively . Addition of TAZ did not make significant change of transferability of PIPC to CSF . Considering the antibacterial effect of TAZ/PIPC against main causative organism of meningitis, this agent was thought to be effective for the treatment of purulent meningitis. Jpn J Antibiot, 1998 Jul, 51(7), 494 - 500 {Resistance to macrolide antibiotics found in methicillin-resistant Japanese clinical isolates of Staphylococcus aureus in 1996}; Nakamura A et al.; Minimum inhibitory concentrations (MICs) of erythromycin, clarithromycin, roxithromycin, oleandomycin, triacetyloleandomycin, azithromycin, josamycin and midecamycin were investigated using 200 strains of methicillin-resistant Staphylococcus aureus (MRSA) clinically isolated in Japan during 1996 . The results show that the MRSAs could be classified into five groups according to MIC patterns to various macrolides and that more than 88% of the strains used were highly-resistant to all macrolides tested . It was found that 9.0% of the strains examined showed a unique MIC pattern different to that of macrolide-lincosamide-streptogramin B antibiotic resistance type . This group was found to be highly resistant to 14-membered but susceptible to 16-membered macrolides . The resistance induction by erythromycin or oleandomycin was observed to increase for clarithromycin and roxithromycin resistances in a part of strains used . On the other hand, for azithromycin, such induction was not observed. Ned Tijdschr Geneeskd, 1998 Jun 20, 142(25), 1425 - 9 {Pain in the hip area accompanied by a fever}; Veldman BA et al.; In 3 patients, 2 women aged 16 and 64 years and 1 man aged 64 years, with pain in the left hip region and fever, the diagnosis psoas abscess was made . After antibiotic treatment and drainage they recovered well . The primary from of psoas abscess is presumably caused by haematogenous spread of bacteria, mostly Staphylococcus aureus . The secondary form is caused by spread of infection from surrounding tissue, mostly gastrointestinal micro-organisms with Crohn's disease and diverticulitis . Painful passive extension and endorotation as well as a painful flexion stress-test of the hip joint can indicate a psoas abscess . Echography and blood cultures should be performed if a psoas abscess is suspected . If echography is inconclusive, CT-scan can establish the diagnosis . The psoas abscess should be treated by percutaneous or surgical drainage combined with antibiotic therapy . The underlying cause of a secondary psoas abscess should be treated separately. Ann Fr Anesth Reanim, 1997, 16(8), 964 - 6 {Peridural abscess complicating spinal anesthesia in a diabetic patient}; Popesco D et al.; Infectious complications of spinal or epidural anaesthesia are rare, particularly after spinal anaesthesia . Most of them consist of a meningitis . We report a case of epidural abscess due to Staphylococcus aureus following spinal anaesthesia in a 62-year-old diabetic patient, diagnosed 45 days after the puncture with bacterial samples and magnetic resonance imaging . The pejorative neurological outcome required a laminectomy in spite of an efficient anti-staphylococcal treatment. Lett Appl Microbiol, 1998 Aug, 27(2), 98 - 100 Anti-MRSA activity of sophoraflavanone G and synergism with other antibacterial agents; Sakagami Y et al.; Anti-MRSA activity of sophoraflavanone G (SFG) and synergism between SFG and antibacterial agents against MRSA (methicillin-resistant Staphylococcus aureus) were evaluated by means of Minimal Inhibitory Concentrations (MIC) . The MICs of SFG against 27 strains of MRSA ranged from 3.13 to 6.25 micrograms ml-1 . Synergism between SFG and vancomycin hydrochloride (VCM) or fosfomycin (FOM) was observed (the fraction inhibitory concentration (FIC) indices were 0.16 and 0.48), while partial synergism was admitted between SFG and other antibacterial agents such as methicillin (DMPPC), cefzonam (CZON), gentamicin (GM), minocycline (MINO) and levofloxacin (LVFX) (the FIC indices were 0.71, 0.73, 0.69, 0.65 and 0.58, respectively) . These findings suggest that SFG in combination with VCM or FOM may be useful in controlling MRSA infections. J Appl Microbiol, 1998 Sep, 85(3), 537 - 44 Growth and enterotoxin production of Staphylococcus aureus during the manufacture and ripening of Camembert-type cheeses from raw goats' milk; Meyrand A et al.; Tests were carried out to determine the effect of manufacturing procedures for a Camembert-type cheese from raw goats' milk on the growth and survival of Staphylococcus aureus organisms added to milk at the start of the process, and to study the possible presence of staphylococcal enterotoxin A in these cheeses . The initial staphylococcal counts were, respectively, 2, 3, 4, 5 and 6 log cfu ml-1 . Cheese was prepared following the industrial specifications and ripened for 41 d . Detection of enterotoxins was done by the Vidas SET test and by an indirect double-sandwich ELISA technique using antienterotoxin monoclonal antibodies . Generally, numbers of microbes increased at a similar rate during manufacture in all cheeses until salting . During the ripening period, the aerobic plate count population and Staph . aureus levels remained stable and high . There was an approximately 1 log reduction of Staph . aureus in cheeses made with an initial inoculum of Staph . aureus greater than 10(3) cfu ml-1 at the end of the ripening period (41 d) compared with the count at 22 h . The level of staphylococcal enterotoxin A recovered varied from 1 to 3.2 ng g-1 of cheese made with an initial population of 10(3)-10(6) cfu ml-1 . No trace of enterotoxin A was detected in cheeses made with the lowest Staph . aureus inoculum used in this study. Arch Biochem Biophys, 1998 Oct 1, 358(1), 68 - 73 Neuronal nitric oxide synthase-membrane phospholipid interactions; Watanabe Y et al.; Most of the neuronal nitric oxide synthase (nNOS) is present in the particulate fraction of tissue extracts . Here, we show that the calmodulin (CaM)-binding domain of nNOS interacts with anionic phospholipid vesicles but not with neutral ones . Identification of residues in the CaM-binding domain of nNOS as the key domain for the interaction is also documented . Recombinant wild-type nNOS was found to associate with phosphatidylserine (PS) or phosphatidic acid (PA) but not with phosphatidylethanolamine (PE) or phosphatidylcholine (PC), indicating that nNOS-phospholipid binding requires an electrostatic interaction . A synthetic peptide corresponding to residues 732-754 blocked the interaction of nNOS with PS . Furthermore, a purified fusion protein containing residues 724-755 interacted with PS in a competitive fashion with CaM . Inactive nNOS lacking CaM-binding ability, generated by mutation of (Lys732LysLeu) to (Asp732AspGlu) (Watanabe, Y., Hu, Y., and Hidaka, H., FEBS Lett . 403, 75-78, 1997) did not interact with PS . Preincubation of nNOS with PS protected subsequent limited proteolysis of the synthase by Staphylococcus aureus V8 protease, probably as a result of conformational changes in the protein . Wild-type nNOS was found almost entirely in the membrane fraction of Sf9 cells, whereas inactive nNOS was also found in cytosolic fraction in Sf9 cells expressing the mutant enzyme . These results demonstrate that the mutated hydrophobic/basic amino acid cluster in nNOS sequence, Lys732LysLeu, is essential for nNOS-PS and nNOS-CaM interactions . Arch Mal Coeur Vaiss, 1998 Jun, 91(6), 753 - 7 {Infections secondary to implantation of cardiac pacemakers}; Da Costa A et al.; Infectious complications of pacemaker implantation are not common but may be particularly severe . Localised wound infections at the site of implantation have been reported in 0.5% of cases in the most recent series with an average of about 2% . The incidence of septicaemia and infectious endocarditis is lower, about 0.5% of cases . The operator's experience, the duration of the procedure and repeat procedures are considered to be predisposing factors . The main cause of these infections is though to be local contamination during the implantation . The commonest causal organism is the staphylococcus (75 to 92%), the staphylococcus aureus being the cause of acute infections whereas the staphylococcus epidermis is associated with cases of secondary infection . The usual clinical presentation is infection at the site of the pacemaker but other forms such as abscess, endocarditis, rejection of the implanted material, septic emboli and septic phlebitis have been described . The diagnosis is confirmed by local and systemic biological investigations and by echocardiography (especially transoesophageal echocardiography) in cases of right heart endocarditis . There are two axes of treatment: bactericidal double antibiotherapy and surgical ablation of the infected material either percutaneously or by cardiotomy . Though controversial, and unsupported by scientific evidence, the role of systematic, preoperative, prophylactic antibiotic therapy in the prevention of these complications seems to be increasing. Arch Mal Coeur Vaiss, 1998 Jul, 91(7), 893 - 7 {Communication between the left ventricle and the right atrium in infectious endocarditis . Diagnosis using Doppler-echocardiography}; Grand A et al.; The diagnosis of a communication between the left ventricle and right atrium was made by transthoracic and transoesophageal echocardiography in a 67 year old man with a recurrence of a methicillin-resistant staphylococcus aureus infectious endocarditis complicating aortic valve replacement with a bioprosthesis seven weeks previously . This diagnosis was confirmed at surgery; the left ventricular-right atrial communication was closed by suturing its edges and a new aortic valve prosthesis was implanted . Unfortunately, the patient died 4 months later of myocardial dysfunction although the infectious endocarditis seemed to have been sterilised by antibiotic therapy . Doppler echocardiography, especially using the transoesophageal approach is the best diagnostic method for rare complications of infectious endocarditis, usually of the aortic valve, the prognosis of which is improved by early surgery and appropriate antibiotic therapy for the causal organisms. Br J Dermatol, 1998 Jun, 138(6), 1036 - 8 Phagocytosis and oxidative burst by neutrophils in patients with recurrent furunculosis; Demircay Z et al.; Neutrophil phagocytosis of fluorescently labelled Staphylococcus aureus and oxidative burst by the neutrophils were assessed by flow cytometry in 22 patients with recurrent furunculosis and in 17 controls . Phagocytosis and oxidative burst were not found to be significantly different between the patients and controls . Low serum iron concentrations were demonstrated in six patients (27%) . In these patients with hypoferraemia, oxidative burst was significantly lower than in the patients without hypoferraemia and in the controls . These data suggest that hypoferraemia may be an important predisposing factor in a subgroup of patients with recurrent furunculosis in impairing oxidative killing by neutrophils. Br J Dermatol, 1998 Jun, 138(6), 1022 - 9 Flucloxacillin in the treatment of atopic dermatitis; Ewing CI et al.; Although colonization of atopic dermatitis by Staphylococcus aureus is universal and bacterial infection is common, it is not known whether antibiotic therapy is helpful in eczematous children who do not have any signs suggestive of bacterial infection . Fifty children aged 1-16 years with atopic dermatitis took part in a randomized double-blind placebo-controlled study of 4 weeks treatment with oral flucloxacillin, with an 8-week follow-up period . The change in the mean of the log10 of the counts/cm2 of S . aureus after 4 weeks of treatment was significantly different for patients receiving treatment, compared with the change for those receiving the placebo (P = 0.008) . However, the difference in the change at 14 days after stopping treatment was not significant (P = 0.32) . Methicillin-resistant strains of S . aureus were cultured from five children during or after treatment . Flucloxacillin did not improve the symptoms or clinical appearance of atopic dermatitis and only temporarily changed skin colonization by S . aureus. Anasthesiol Intensivmed Notfallmed Schmerzther, 1998 Aug, 33(8), 497 - 500 {Methicillin-reistant Staphylococcus aureus: risk factors for infection-colonization and clonal heterogeneity in intensive care units}; Wichelhaus TA et al.; PURPOSE: The aim of this study was to determine the risk factors associated with colonisation/infection by methicillin-resistant Staphylococcus aureus (MRSA) and to demonstrate the chain of infections by genotyping of all MRSA isolates . METHODS: A total of 6143 microbiological samples from 1753 patients was obtained at a surgical ICU of Frankfurt University hospital during 1995 . RESULTS: MRSA was detected in 1.6% of patients and three members of staff (3.3%) . Typing of these 31 MRSA-strains (first isolates) by macrorestriction analysis of chromosomal DNA revealed nine different genotypes . More than 60% of all isolates belonged to one type that was confirmed to be closely related to the "South Germany" epidemic strain . A strong correlation between severity of underlying disease and length of hospitalisation on the one hand and detection of MRSA on the other could be demonstrated . Detection of MRSA was significantly more common in patients with adult respiratory distress syndrome (ARDS), sepsis, kidney failure, prolong respiratory treatment and in patients with prolonged phases of haemodynamic instability . It appeared that the mortality rate of MRSA-infected patients was higher (28.6%) than the mortality rate of all patients (6.5%) . CONCLUSION: Following of a strict hygiene regime is important to prevent clonal spread of MRSA and especially to protect immunocompromised patients from complicating infections. Caries Res, 1998, 32(6), 441 - 6 Efficacy of sterilisation methods and their effect on enamel demineralisation; Amaechi BT et al.; The aim of this project was to determine the effectiveness of sterilisation methods for dental enamel for use in intra-oral cariogenicity tests, and their possible effect on the degree of demineralisation of enamel . Bovine incisors were cut vertically into five portions and each assigned to one of five groups . Group 1 was used as a control while the other four groups were subjected, respectively, to gamma irradiation ( congruent with 25kGy), steam autoclaving (121 degrees C for 15 min), sodium hypochlorite (12% w/v for 24h) and povidone-iodine (7.5% w/v for 24h) . Total viable counts of microorganisms remaining following sterilisation of the specimens were performed following incubation of the specimens for 24h at 37 degrees C . Caries-like lesions were produced in each specimen using an acidic buffer solution (pH4.5) . Sections were cut from each specimen, ground to 80-microgram thickness, and microradiographed . Mineral loss and lesion depth were quantified using transverse microradiography . Statistical analysis was by ANOVA . Dunnett's and Tukey's tests . Microbial growth (Staphylococcus aureus and bacilli) was observed only in control specimens in both brain heart infusion broth and on blood agar plates . The sterilisation methods affected the enamel surface as follows: gamma irradiation (cream discolouration), NaOCl (bleaching), and povidone-iodine (white spot-like lesion) . Compared with the control, there was no significant difference in mineral loss and lesion depth with any of the groups, but the numerical values of mineral loss and lesion depth in groups can be ranked as follows: gamma irradiation <povidone-iodine <control <autoclave <NaOCl . In conclusion, the four sterilisation methods were all effective to sterilise enamel, but gamma irradiation proved the most acceptable method for enamel to be used in cariogenicity tests having the least adverse effect. Infect Immun, 1998 Oct, 66(10), 4588 - 92 Antigenic determinants of Staphylococcus aureus type 5 and type 8 capsular polysaccharide vaccines; Fattom AI et al.; Bacterial capsular polysaccharides (CP) are carbohydrate polymers comprised of repeating saccharide units . Several of these CP have side chains attached to their backbone structures . The side chains may include O-acetyl, phosphate, sialic acid, and other moieties . Those moieties represent the immunodominant epitopes and the most functional ones . The clinically significant Staphylococcus aureus type 5 CP (CP 5) and type 8 CP (CP 8) are comprised of a trisaccharide repeat unit with one O-acetyl group attached to each repeat unit . The immunogenicity of these