J Bacteriol, 1996 Sep, 178(18), 5464 - 71 Role of mecA transcriptional regulation in the phenotypic expression of methicillin resistance in Staphylococcus aureus; Niemeyer DM et al.; The gene required for methicillin resistance in staphylococci, mecA, encodes the low-affinity penicillin-binding protein 2a (PBP2a) . Transcriptional regulation of mecA is accomplished in some isolates by mecR1 and mecI, cotranscribed chromosomal genes that encode a putative signal transducer and a transcriptional repressor, respectively . Two Staphylococcus aureus strains that have identical mecR1-mecI nucleotide sequences, BMS1 and N315P, both exhibit low-level, heterotypic expression of methicillin resistance and contain no beta-lactamase coregulatory sequences . mecR1-mecI was amplified from BMS1 by PCR and was shown to be functional on a high-copy-number plasmid when introduced into an S . aureus strain with a deleted mecR1-mecI locus . Cloned mecR1-mecI repressed phenotypic expression of methicillin resistance, mecA transcription and PBP2a production and mediated PBP2a induction in response to certain beta-lactam antibiotics . However, mecR1-mecI had different regulatory activities in its native chromosomal location in N315P compared with those in BMS1 . Uninduced mecA transcription was markedly repressed in N315P, and mecI inactivation increased mecA transcription and PBP2a production 5- and 40-fold, respectively . Furthermore, the N315P phenotype changed from low-level, heterotypic resistance with intact mecI to high-level, homotypic resistance in strains with disrupted mecI . In contrast, uninduced BMS1 produced abundant mecA transcript and PBP2a, while the disruption of mecI had no effect on phenotype and little effect on mecA transcription or PBP2a production . Thus, mecI-mediated repression of mecA appears to be dysfunctional in BMS1 because of the presence or absence of additional regulatory cofactors . Furthermore, heterotypic resistance expression in this strain is independent of mecA transcriptional regulation. Crit Care Med, 1996 Sep, 24(9), 1482 - 9 Surface heparinization of central venous catheters reduces microbial colonization in vitro and in vivo: results from a prospective, randomized trial; Appelgren P et al.; OBJECTIVE: To evaluate in vitro and in vivo the efficacy of covalent end point-attached heparin to single-lumen polyurethane central venous catheters in reducing microbial adherence and colonization . DESIGN: In vitro study: A controlled bench study . In vivo study: A prospective, randomized, double-blind, clinical trial . SETTING: Intensive care unit in a 1200-bed teaching hospital . INTERVENTIONS: In vitro study: Adhesion of 17 radiolabeled clinical isolates of Staphylococci to catheters was examined in vitro . In vivo study: The outcome of heparinized and control catheters was compared in vivo in patients receiving long-term parenteral nutrition . Fifty-five adult patients were prospectively, blindly randomized to heparinized or control central venous catheters . The catheters, removed on clinical grounds, were analyzed with semiquantitative and quantitative cultures . Blood cultures were done at catheter removal . MEASUREMENTS AND MAIN RESULTS: In vitro study: Coagulase-negative Staphylococci adhered less in vitro to heparinized catheters than to control catheters (p < .05) . In vivo study: Among 32 central venous catheters, or patients who completed the study, catheter-associated bacteremia or fungemia was observed in five patients in the control group (n = 19) and in no patient with a heparinized catheter (n = 13) (p = .047) . Four of 13 catheters in the heparin group were colonized compared with 14 of 19 in the control group (p = .03) . Coagulase-negative Staphylococci were the most frequent microorganisms in both groups . The numbers of organisms found on colonized catheters were larger in the control group than in the heparin group . CONCLUSIONS: Covalent end point surface heparinization appears to have a great impact on both in vitro and in vivo bacterial colonization of central venous catheters . Such heparinization can be a practical and economical approach to the prevention of catheter-associated bacteremia or fungemia. Scand J Immunol, 1996 Sep, 44(3), 261 - 6 Participation of V beta 4(+)-, V beta 7(+)-, and V beta 11(+)-T lymphocytes in haematogenously acquired Staphylococcus aureus nephritis; Verba V et al.; The most frequent pathogen in haematogenously acquired kidney infections in humans in Staphylococcus aureus . In order to characterize in situ the immunological patterns of septic nephritis we developed a murine model of this disease . A single intravenous injection of S . aureus producing toxic shock syndrome toxin-1 resulted in high frequency of inflammatory kidney lesions . Histopathologically, both focal and diffuse inflammatory infiltrates were seen in kidney cortex and medulla . Immunohistochemical evaluation revealed high numbers of Mac-1+ phagocytic cells as well as CD4(+)-and CD8(+)-lymphocytes . The expansion of lymphocytes carrying T-cell receptor V beta chain 4, 7, and 11 families in the kidney was observed . Our results suggest that the haematogenously acquired kidney infection by superantigen-producing staphylococci leads to migration, in situ activation and expansion of responding T-lymphocyte subsets. J Med Chem, 1996 Aug 30, 39(18), 3596 - 600 The chemistry of pseudomonic acid . 17 . Dual-action C-1 oxazole derivatives of pseudomonic acid having an extended spectrum of antibacterial activity; Broom NJ et al.; A series of C-1 oxazole isosteres of pseudomonic acid A (mupirocin) bearing a nitroheterocycle have been synthesized, and significant differences in both spectrum of activity and potency were found between these derivatives and mupirocin . Additionally, the antibacterial potency of two members of this class of compounds against mupirocin-resistant staphylococci could not be accounted for solely by inhibition of the target enzyme isoleucyl-tRNA synthetase (IRS), indicating an additional mode of action . The most potent compound, the nitrofuran 3f (SB 205952), was the most electron affinic derivative prepared and was transformed by NAD(P)H-dependent bacterial reductases at a rate similar to that for nitrofurantoin . The second mode of action of this compound may therefore arise from its reduction to a species with cellular targets other than IRS . In in vivo studies, 3f was shown to be a very effective agent by both the subcutaneous and oral routes of administration. Int J Food Microbiol, 1996 Aug, 31(1-3), 325 - 31 Identification and ribotyping of Staphylococcus xylosus and Staphylococcus equorum strains isolated from goat milk and cheese; Meugnier H et al.; Twenty-five strains of staphylococci isolated from goat milk and cheese were identified as belonging to the Staphylococcus xylosus/equorum group using the ID 32 Staph system (bioMerieux, Marcy-L'Etoile, France) . This system, however, was not able to discriminate between these two species for 19 of the strains tested . Ribotyping was performed on these 25 strains, as well as on three reference strains of each of these two species . Hybridization membranes were scanned and analyzed using the Taxotron software package (Taxolab, Institut Pasteur, Paris, France) . A dendrogram representation showed that ribotypes were distributed in two clear-cut clusters corresponding to S . equorum (21 strains) and S . xylosus (four strains). J Antimicrob Chemother, 1996 Aug, 38(2), 259 - 63 In-vitro activity of ofloxacin, levofloxacin and D-ofloxacin against staphylococci; von Eiff C et al.; The in-vitro activity of levofloxacin was compared with ofloxacin and D-ofloxacin against 130 isolates of Staphylococcus aureaus and 117 isolates of coagulase-negative staphylococci, using the agar dilution method . In general, levofloxacin was equally active or up to fourfold more active than ofloxacin against all staphylococci, including 61 methicillin-resistant S . aureus . In contrast, D-ofloxacin showed little activity against all isolates tested. Eur J Pediatr, 1996 Aug, 155 Suppl 2, S21 - 4 New modalities for treating neonatal infection; Bedford-Russell AR; While the overall incidence of infection has remained constant at approximately 7/1000 livebirths, the last decade has witnessed a reduction in early onset infections and a relative increase in nosocomial sepsis, chiefly with coagulase-negative staphylococci . Immaturity of host defence mechanisms contributes to an increasing susceptibility to infection with decreasing gestational age and birth weight . In the past, efforts to enhance host defence have included the use of granulocyte infusions, fresh frozen plasma, exchange blood transfusions and immunoglobulin therapy . Current trials are investigating the use of agents which enhance endogenous defence mechanisms, such as, recombinant human granulocyte colony-stimulating factant and recombinant human granulocyte-macrophage colony-stimulating factor and of pentoxifylline . In the meantime strict attention to handwashing and aseptic technique remain the best methods of preventing nosocomial sepsis. Chest Surg Clin N Am, 1996 Aug, 6(3), 419 - 38 Current diagnostic methods and medical management of thoracic empyemas; Lee-Chiong TL Jr et al.; Infections can invade the normally sterile pleural space leading to the development of parapneumonia effusions or empyemas . Pneumonia, thoracic surgery, and trauma, together, are responsible for most cases of empyemas . Pneumococci and staphylococci remain the predominant etiologic organisms . In addition, gram-negative aerobic bacteria and anaerobes are emerging as important pathogens . Most parapneumonic pleural effusions, regardless of their underlying etiology and bacteriology, evolve through definable stages . For each stage of the disease, specific therapeutic measures, either medical or surgical, are available. J Bacteriol, 1996 Aug, 178(16), 4975 - 83 Identification and characterization of the origin of conjugative transfer (oriT) and a gene (nes) encoding a single-stranded endonuclease on the staphylococcal plasmid pGO1; Climo MW et al.; The genes mediating the conjugative transfer of the 52-kb staphylococcal plasmid pGO1 are within a 14.4-kb gene cluster designated trs . However, a clone containing trs alone cannot transfer independently and no candidate oriT has been found within or contiguous to trs . In this study, we identified a 1,987-bp open reading frame (ORF) 24 kb 3' and 13 kb 5' to trs that was essential for conjugative transfer: transposon insertions into the ORF abolished transfer and a plasmid containing the ORF could complement these transposon-inactivated pGO1 mutants for transfer . Analysis of the nucleotide sequence of this ORF revealed significant homology between the amino terminus of its predicted protein and those of several single-stranded endonucleases . In addition, a 12-bp DNA sequence located 100 bp 5' to the ORF's translational start site was identical to the oriT sequences of the conjugative or mobilizable plasmids RSF1010, pTF1, R1162, pSC101, and pIP501 . The ability of the ORF, designated nes (for nicking enzyme of staphylococci), to generate a single-stranded nick at the oriT was demonstrated in Escherichia coli by alkaline gel and DNA sequence analysis of open circular plasmid DNA . Plasmids that could be converted to the open circular form by the presence of oriT and nes could also be mobilized at high frequency into Staphylococcus aureus recipients with a second plasmid containing only trs . We propose that the 14.4 kb of trs and the approximately 2.2 kb of the oriT-nes region, coupled with an origin of replication, make up the minimal staphylococcal conjugative replicon. Infect Immun, 1996 Aug, 64(8), 3425 - 8 Ingestion of Staphylococcus aureus, Staphylococcus epidermidis, and Escherichia coli by human peritoneal mesothelial cells; Visser CE et al.; In the present study we examined whether mesothelial cells can ingest and digest bacteria . The results showed that all strains were ingested . Ingested staphylococci proliferated abundantly, and only a few were digested . Escherichia coli, however, was digested during the first 8 h, whereafter the mesothelial cells disintegrated and proliferation of bacteria could be observed . The clinical implications of these findings are discussed. J Bacteriol, 1996 Aug, 178(15), 4696 - 703 Peptidoglycan synthesis and structure in Staphylococcus haemolyticus expressing increasing levels of resistance to glycopeptide antibiotics; Billot-Klein D et al.; The structures of cytoplasmic peptidoglycan precursor and mature peptidoglycan of an isogenic series of Staphylococcus haemolyticus strains expressing increasing levels of resistance to the glycopeptide antibiotics teicoplanin and vancomycin (MICs, 8 to 32 and 4 to 16 microg/ml, respectively) were determined . High-performance liquid chromatography, mass spectrometry, amino acid analysis, digestion by R39 D,D-carboxypeptidase, and N-terminal amino acid sequencing were utilized . UDP-muramyl-tetrapeptide-D-lactate constituted 1.7% of total cytoplasmic peptidoglycan precursors in the most resistant strain . It is not clear if this amount of depsipeptide precursor can account for the levels of resistance achieved by this strain . Detailed structural analysis of mature peptidoglycan, examined for the first time for this species, revealed that the peptidoglycan of these strains, like that of other staphylococci, is highly cross-linked and is composed of a lysine muropeptide acceptor containing a substitution at its epsilon-amino position of a glycine-containing cross bridge to the D-Ala 4 of the donor, with disaccharide-pentapeptide frequently serving as an acceptor for transpeptidation . The predominant cross bridges were found to be COOH-Gly-Gly-Ser-Gly-Gly-NH2 and COOH-Ala-Gly-Ser-Gly-Gly-NH2 . Liquid chromatography-mass spectrometry analysis of the peptidoglycan of resistant strains revealed polymeric muropeptides bearing cross bridges containing an additional serine in place of glycine (probable structures, COOH-Gly-Ser-Ser-Gly-Gly-NH2 and COOH-Ala-Gly-Ser-Ser-Gly-NH2) . Muropeptides bearing an additional serine in their cross bridges are estimated to account for 13.6% of peptidoglycan analyzed from resistant strains of S . haemolyticus . A soluble glycopeptide target (L-Ala-gamma-D-iso-glutamyl-L-Lys-D-Ala-D-Ala) was able to more effectively compete for vancomycin when assayed in the presence of resistant cells than when assayed in the presence of susceptible cells, suggesting that some of the resistance was directed towards the cooperativity of glycopeptide binding to its target . These results are consistent with a hypothesis that alterations at the level of the cross bridge might interfere with the binding of glycopeptide dimers and therefore with the cooperative binding of the antibiotic to its target in situ . Glycopeptide resistance in S . haemolyticus may be multifactorial. Ugeskr Laeger, 1996 Jul 29, 158(31), 4403 - 5 {Infections in connection with epidural catheterization}; Holt HM et al.; Seventy-eight patients with culture-positive epidural catheters were studied . Fifty-nine had symptoms of exit site infection and 11 patients had clinical meningitis, two of whom also had en epidural abscess . This corresponds to a local infection incidence of at least 4.3% and an incidence of central nervous system infection of at least 0.7% at Odense University Hospital . The patients with generalized symptoms of infection had been catheterized for a longer time, and were older than patients with only local symptoms of infection . The microorganisms isolated from the epidural catheters were coagulase- negative staphylococci (41%), Staphylococcus aureus (35%), Gram-negative bacilli (14%) and other bacteria (10%) . The Gram-negative bacilli and S . aureus caused serious infections more frequently than the others . We discuss the symptoms and diagnosis of spinal epidural abscess and propose prophylactic and diagnostic guidelines for epidural catheter-related infections. FEBS Lett, 1996 Jul 29, 390(3), 327 - 33 Surface display on staphylococci: a comparative study; Robert A et al.; Two different host-vector expression systems, designed for cell surface display of heterologous receptors on Staphylococcus xylosus and Staphylococcus carnosus, respectively, were compared for the surface display of four variants of a 101 amino acid region derived from the G glycoprotein of human respiratory syncytial virus (RSV) . Surface localization of the different chimeric receptors was evaluated by a colorimetric assay and by fluorescence-activated cell sorting . It was concluded that the S . carnosus system was better both in the ability to translocate inefficiently secreted peptides and in the number of exposed hybrid receptors . The potential use of the described staphylococci as live bacterial vaccine vehicles or alternatives to filamentous phages for surface display of protein libraries is discussed. Int J Cardiol, 1996 Jul 26, 55(2), 193 - 7 Successful treatment of Staphylococcus lugdunensis endocarditis complicated by multiple emboli: a case report and review of the literature; Koh TW et al.; We describe the prompt diagnosis and successful treatment of mitral valve endocarditis in a 52-year-old woman due to a recently described coagulase negative staphylococcus, Staphylococcus lugdunensis . Endocarditis due to this organism has a high mortality rate with 8 out of 12 published cases ending in fatality . Review of the literature revealed that use of commercial identification systems can lead to misidentification of Staphylococcus lugdunensis and consequently delay appropriate treatment . It is clinically important to distinguish Staphylococcus lugdunensis from other coagulase negative staphylococci by detailed microbiological testing . The awareness of this from of endocarditis and its natural history is important since it differs significantly from other coagulase negative staphylococci . It highlights the need for early surgical intervention not only for haemodynamic complications but also for recurrent emboli . Multiple emboli appears to be a frequent finding on review of the literature. Arch Intern Med, 1996 Jul 8, 156(13), 1470 - 4 Pneumococcal pyomyositis . Case report, review of the literature, and comparison with classic pyomyositis caused by other bacteria; Collazos J et al.; Pyomyositis is caused by staphylococci in 70% to 90% of patients . We report a case of pneumococcal pyomyositis (PP), review the 11 cases previously published, and compare the features of pneumococcal pyomyositis with those of classic (nonpneumococcal) pyomyositis . Several clinical characteristics have been identified that are notably different in both groups . Psoas muscle involvement was observed in two thirds of the patients with PP, and a source for the infection was identified in half of the patients . Patients with PP were older than those with classic pyomyositis . Men were affected less often than women with PP, but the opposite was the rule in classic pyomyositis . The systemic response to the infection was more prominent in patients infected with pneumococci than from other causes . Most patients with PP were successfully treated with antibiotics and drainage . Secondary meningitis was observed in 3 patients with psoas muscle abscess caused by pneumococci . Mortality is low in pyomyositis regardless of the causative pathogen. Biochemistry, 1996 Jul 2, 35(26), 8686 - 95 Regiospecificity of aminoglycoside phosphotransferase from Enterococci and Staphylococci (APH(3')-IIIa); Thompson PR et al.; The broad-spectrum aminoglycoside phosphotransferase, APH(3')-IIIa, confers resistance to several aminoglycoside antibiotics in opportunistic pathogens of the genera Staphylococcus and Enterococcus . The profile of the drug resistance phenotype suggested that the enzyme would transfer a phosphate group from ATP to the 3'-hydroxyl of aminoglycosides . In addition, resistance to the 3'-deoxyaminoglycoside antibiotic, lividomycin A, suggested possible transfer to the 5"-hydroxyl of the ribose {Trieu-Cuot, P., & Courvalin, P . (1983) Gene 23, 331-341} . Using purified overexpressed enzyme, we have prepared and purified the products of APH(3')-IIIa-dependent phosphorylation of several of aminoglycoside antibiotics . Mass spectral analysis revealed that 4,6-disubstituted aminocyclitol antibiotics such as amikacin and kanamycin are monophosphorylated, while 4,5-disubstituted aminoglycosides such as butirosin A, ribostamycin, and neomycin B are both mono- and diphosphorylated by APH(3')-IIIa . Using a series of one- and two-dimensional 1H, 13C, and 31P NMR experiments, we have unambiguously assigned the regiospecificity of phosphoryl transfer to several antibiotics . The 4,6-disubstituted aminocyclitol antibiotics are exclusively phosphorylated at the 3'-OH hydroxyl, and the 4,5-disubstituted aminocyclitol antibiotics can be phosphorylated at both the 3'- and 5"-hydroxyls . The first phosphorylation can occur on either the 3'- or 5"-hydroxyl group of neomycin B or butirosin A . Initial phosphotransfer to the 3'-position predominates for butirosin while the 5"-OH is favored for neomycin . These results open the potential for the rational design of aminoglycoside kinase inhibitors based on functionalization of either the 6-aminohexose or the pentose rings of aminoglycoside antibiotics. Biochemistry, 1996 Jul 2, 35(26), 8680 - 5 Catalytic mechanism of enterococcal kanamycin kinase (APH(3')-IIIa): viscosity, thio, and solvent isotope effects support a Theorell-Chance mechanism; McKay GA et al.; Bacterial resistance to the aminoglycoside antibiotics is manifested primarily through the production of enzymes which covalently modify these drugs . The Enterococci and Staphylococci produce an ATP-dependent kinase, APH(3')-IIIa, which phosphorylates such antibiotics as kanamycin, amikacin, and neomycin, and this enzyme shows a Theorell-Chance kinetic mechanism by traditional product and analogue inhibitor analysis and by the alternative substrate diagnostic {McKay, G . A., & Wright, G . D . (1995) J . Biol . Chem . 270, 24686-24692} . We report that the APH(3')-IIIa exhibits small solvent (VH/VD approximately equal to 1.50) and thio effects (VATP/VATP gamma S = 2) indicating hydroxyl group deprotonation and nucleophilic attack on ATP do not significantly contribute to the overall steady-state rate . The enzymatic rates were determined with the viscogens PEG 8000, glycerol, and sucrose, and these experiments demonstrate that ATP binding and ADP release are diffusion controlled and that ADP release is solely rate limiting for APH(3')-IIIa . In addition, the slope of V/K for ATP vs relative viscosity is greater than the theoretical limit of 1, suggesting a possible enzyme conformational change upon binding of ATP . This new experimental evidence supports a Theorell-Chance mechanism for APH(3')-IIIa. Zh Mikrobiol Epidemiol Immunobiol, 1996 Jul-Aug, (4), 30 - 3 {The taxonomic importance of the capacity of bacteria in the genus Staphylococcus for the inactivation of a number of natural anti-infectious resistance factors}; Bukharin OV et al.; Bacteria of the species S . aureus are characterized by a wide range of signs indicating the inactivation of natural resistance factors, including anticomplement activity, which makes them different from coagulase-negative staphylococci exhibiting only antilysozyme activity . The taxonomic importance of these tests and their possible use for the identification of staphylococcal species are discussed. Mycoses, 1996 Jul-Aug, 39(7-8), 313 - 21 Characteristics of Malassezia pachydermatis strains isolated from canine otitis externa; Kiss G et al.; The morphological, cultural and biochemical characteristics of 80 M . pachydermatis strains isolated from cases of canine otitis externa were studied . Microscopically, the strains could be subdivided into two phenotypes . All M . pachydermatis strains grew well on Sabouraud glucose, yeast morphology and modified malt extract agar, but formed two distinct colony types . All strains were characterized by no fermentation . Assimilation of glucose, mannitol (42 strains), sorbitol (40 strains) and peptone was observed, but no ethanol assimilation . Urease and catalase tests were positive, while indole and acetoin production was not detected . All strains showed proteinase, caseinase, lecithinase and peroxidase positivity but to varying extents . Esterase activity was observed for all Malassezia strains when using Tween 20, 40 and 60, whereas Tween 80 was hydrolysed by only 42 strains . No coagulase or haemagglutinating activities were detected . When compared for satellite phenomenon and vitamin requirements, some Malassezia strains could not grow in the absence of nicotinic acid but grew well in the presence of staphylococci . In susceptibility tests, all strains showed the highest susceptibility to ketoconazole . On the basis of the biochemical differences, M . pachydermatis seems to be a heterogeneous species and can be divided into two groups. Klin Lab Diagn, 1996 Jul-Aug, (4), 53 - 4 {Micromethod of determining phosphatase activity of staphylococci}; Sboichakov VB et al.; A rapid method for measuring phosphatase activity of staphylococci has been developed . Russian 96-well polystyrene plates are used . 0.1 ml of 0.2% sodium phenolphthalein phosphate solution in 1% peptone water, pH 6.8 to 7.0, and 0.1 ml of the tested strain suspension in normal saline are put into wells and incubated for 5 h at 37 degrees C . The results are assessed after adding 0.05 ml of 2.5% ammonia spirit into each well . If the result is positive, the medium is colored raspberry-red, if negative-light-yellow . The method is simple, available, and cheap. Diagn Microbiol Infect Dis, 1996 Jul, 25(3), 107 - 12 Phenotypic detection of mec A-positive staphylococcal blood stream isolates: high accuracy of simple disk diffusion tests; Cormican MG et al.; Detection of oxacillin-resistance in staphylococci by phenotypic methods remains problematic . Although standardized susceptibility test methods are adequate for Staphylococcus aureus, many are less satisfactory for the coagulase-negative staphylococci (CNS) . We have studied 108 consecutive blood culture isolates of staphylococci . The mec A gene was detected by PCR in one S . aureus and 55 CNS isolates . Susceptibility testing was performed as follows: oxacillin (1-microgram), ceftizoxime (30-microgram), and cephalothin (30-microgram) by disk diffusion; oxacillin, ceftizoxime, cephalothin, methicillin, ampicillin, ampicillin/ sulbactam, penicillin, cefazolin, imipenem, and meropenem by the broth microdilution method . In addition, isolates were tested by the oxacillin agar screen plate method . The single oxacillin-resistant S . aureus strain was detected by all oxacillin susceptibility test methods and by the ceftizoxime disk and MIC methods . Two oxacillin-susceptible S . aureus were intermediate (minor error) by ceftizoxime broth microdilution (MIC, 16 micrograms/mL) . The most sensitive, simple phenotypic methods for detection of oxacillin-resistant CNS (mec A positive) were as follows: oxacillin disk diffusion at 98%, oxacillin screen plate at 91%, oxacillin broth microdilution at 87%, ceftizoxime disk diffusion at 100%, ceftizoxime broth microdilution at 87%, and methicillin broth microdilution at 83% . These results indicate that oxacillin and ceftizoxime disk diffusion tests are the most accurate phenotypic methods in routine clinical use for detection of oxacillin-resistant CNS . Oxacillin broth microdilution MIC testing (2% NaCl supplement) would perform more satisfactorily (100% sensitivity) with an adjusted interpretive breakpoint at < or = 0.5 microgram/mL, in contrast to the lower accuracy of the "so-called" reference agar screen test. Eur J Clin Microbiol Infect Dis, 1996 Jul, 15(7), 567 - 73 Comparison of disk diffusion, the E test, and detection of mecA for determination of methicillin resistance in coagulase-negative staphylococci; Mulder JG; The aim of this study was to find a reliable, fast, and simple alternative to the methicillin disk method for determination of methicillin resistance in coagulase-negative staphylococci, since results of this method are often difficult to read due to growth within the zone of inhibition . The sensitivity of 319 strains of coagulase-negative staphylococci to a 5 microgram methicillin disk on Mueller-Hinton agar using an incubation period of 48 h was compared with that of 1 microgram and 5 micrograms oxacillin disks on Mueller-Hinton agar with or without 2% NaCl, using an incubation period of 24 h . The detection of mecA (MecAgen) by the polymerase chain reaction was used as a standard . Minimum inhibitory concentrations were determined by means of the E test . Of the 225 mecA-positive strains, 190, 215, and 193 were resistant to 5 micrograms methicillin, 1 microgram oxacillin and 5 micrograms oxacillin disks on Mueller-Hinton agar, respectively, and 216, 218, and 223 were resistant on Mueller-Hinton agar with 2% NaCl . Of the 94 mecA-negative strains, 89, 93, and 94 were susceptible to 5 micrograms methicillin, 1 microgram oxacillin, and 5 micrograms oxacillin disks on Mueller-Hinton agar, respectively, and 92, 93, and 94 were susceptible on Mueller-Hinton agar with 2% NaCl . Using breakpoints of 2 micrograms/ml for oxacillin resistance and 8 micrograms/ml for methicillin resistance, the E test yielded sensitivities of 99.6 and 99.1% and specificities of 97.9 and 98.9% after 48 h of incubation . The 5 microgram oxacillin disk was faster and easier to read than the methicillin disk and correlated better with detection of mecA than the methicillin disk of the 1 microgram oxacillin disk. Int J Food Microbiol, 1996 Jul, 30(3), 373 - 8 Identification of micrococcaceae isolated from goat's milk and cheese in the Poitou-Charentes region; Vernozy Rozand C et al.; One hundred and ninety strains of coagulase-negative staphylococci were isolated from goat's milk, whey and cheese at various stages of manufacture . Sixteen different coagulase negative Staphylococci (CNS) species were recovered, 3 of which were predominant: Staphylococcus simulans, Staphylococcus epidermidis and Staphylococcus xylosus . The prevalent species were recovered at least at two different stages of cheese manufacturing, suggesting a better adaptation to the environment . After 15 days of ripening, the cheeses showed lower counts of Micrococcaceae. Int J Food Microbiol, 1996 Jul, 30(3), 271 - 80 Enterotoxin production by coagulase-negative staphylococci isolated from goats' milk and cheese; Vernozy-Rozand C et al.; An antigen related to the Enterotoxin E from Staphylococcus aureus was produced by ten of 187 coagulase-negative staphylococci (CNS) isolated from goats' milk, whey and cheese in quantities ranging from 10 to 90 ng/ml supernatant . The enterotoxin-producing strains were identified at the species level as S . simulans, S . xylosus, S . equorum, S . lentus and S . capitis . Detection of the enterotoxins was done by the VIDAS SET test (bioMerieux) and by an indirect double-sandwich ELISA technique using anti-enterotoxin monoclonal antibodies . The results obtained were further confirmed by Southern blotting, using two radioactive oligonucleotide probes that hybridized specifically with the gene of S . aureus coding for the enterotoxin E. Infect Control Hosp Epidemiol, 1996 Jul, 17(7), 419 - 22 The effect of a portable HEPA-filtered body exhaust system on airborne microbial contamination in a conventional operating room; Bohn WW et al.; OBJECTIVE: To study the effect of a portable HEPA-filtered air exhaust system (Stackhouse Freedom Surgical Helmet System) on airborne microbial contamination in a modern conventional operating room . DESIGN AND SETTING: Microbial air sampling was done with a two-stage Anderson sampler at the wound site during 46 total joint replacements . All operations were performed by the same surgeon in the same operating room at a large community hospital . RESULTS: In 18 cases done without air exhaust hoods, the number of bacterial and fungal colony-forming units (CFU) ranged from 0.6 to 11.7 (mean, 3.6) . Air sampling during 28 operations with the operating team in air exhaust hoods revealed a mean of 3.6 CFU (range, 0 to 11.4) . Bacterial CFU averaged 3.4 without hoods and 3.2 with exhaust hoods . Coagulase-negative staphylococci were the most common isolates (48% of isolates with hood, 55% without hood) . No infections occurred . CONCLUSION: We concluded that these air exhaust hoods did not lower airborne microbial contamination detectable with this air sampling method, as compared to standard head cover and mask, in a modern conventional operating room. Zentralbl Bakteriol, 1996 Jul, 284(2-3), 390 - 401 Drug delivery concepts for the efficacious prevention of foreign-body infections; Schierholz JM et al.; Several classes of antibiotics released from polyurethanes and silicones were examined for their activity against foreign body colonization by coagulase-negative staphylococci . Beta-lactams, gyrase inhibitors, aminoglycosides, macrolides and rifampicin were used as antimicrobials to impregnate or load the polymers . Coating of polymeric surfaces by precipitation or adsorption techniques lead to a lower and shorter drug release as compared to polymeric devices with incorporated antibiotics . Prolonged drug delivery of matrix-loaded polymers exceeding the microenvironmental minimal bactericidal concentrations (mMBC), ensured the prevention of bacterial colonization . In this study, we have been able to demonstrate the usefulness of a reproducible long-time antimicrobial dosage regime from the internal phase of the implant as compared to surface coated polymers . In addition, pharmacodynamic aspects and the potential of bonded antibiotics for inducing adverse effects such as resistance development and allergic reactions are discussed. Zentralbl Bakteriol, 1996 Jul, 284(2-3), 318 - 28 Effects of the lantibiotic mersacidin on the morphology of staphylococci; Molitor E et al.; Mersacidin is a lanthionine-containing peptide antibiotic (lantibiotic), able to inhibit the growth of a number of Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA) in a manner similar to, but distinct from, vancomycin . In order to further understand the mode of action of this lantibiotic, Staphylococcus simulans 22 cells were treated either with the antibiotics penicillin, tunicamycin or vancomycin or with mersacidin and then compared with untreated cells after electron microscopic examination . Mersacidin treatment brought about a time-dependent, generalised decrease in the thickness of the bacterial cell wall . In addition, mersacidin treatment caused a roughening of the cell wall surface layer and also reduced the thickness and frequency of formation of dividing cell septa . Reduction of cell wall thickness appears to result from inhibition of new wall biosynthesis combined with cell wall turnover . These features of mersacidin-induced effects on cell morphology confirm that it has a novel mode of action (Brotz, H., G . Bierbaum, A . Markus, E . Molitor, and H.-G . Sahl: Antimicrob . Agents Chemother . 39 {1995} 714-719), probably directed towards a membrane-bound biosynthetic step but not towards a specific penicillin-binding-protein. Zentralbl Bakteriol, 1996 Jul, 284(2-3), 297 - 301 In-vitro activity of penicillin G plus sulbactam in comparison with other beta-lactamase inhibitor combinations and oxacillin against staphylococci; Jansen B et al.; The in-vitro activity of penicillin G in combination with the beta-lactamase inhibitor, sulbactam, against penicillin-sensitive S . aureus (n = 10) and penicillin-resistant, methicillin-sensitive S . aureus (n = 69) and S . epidermidis (n = 20) was tested in comparison with ampicillin/sulbactam, amoxicillin/clavulanic acid, piperacillin/tazobactam and oxacillin . The combination of penicillin G plus sulbactam was found to lead to MIC values for beta-lactamase producing staphylococci comparable to those for penicillin-sensitive staphylococci, with MIC90 values between < or = 0,03 mg/L and 0.06 mg/L. Zentralbl Bakteriol, 1996 Jul, 284(2-3), 285 - 96 Isolation of a new epidermin variant from two strains of Staphylococcus epidermidis--frequency of lantibiotic production in coagulase-negative staphylococci; Israil AM et al.; A novel lantibiotic identified during the screening of 187 coagulase-negative staphylococci, has been structurally characterised . (Val1, Leu6)-epidermin was purified from culture supernatants of two Staphylococcus epidermidis strains, BN-V1 and BN-V301 following successive separation by adsorption, cation exchange and C18 reversed-phase chromatography . Separation of the purified peptides by SDS-polyacrylamide gel electrophoresis suggested a molecular mass of ca . 2000 Da and electrospray mass spectrometry subsequently demonstrated that both peptides had a mass of 2151 Da; a mass which is different from that of all previously described antimicrobial peptides . Amino acid analysis showed that both peptides contained lanthionine and were very similar to epidermin, while automated N-terminal sequencing by Edman degradation confirmed the identity of the two peptides as a natural variant of the antimicrobial lantibiotic epidermin, and demonstrated the conservative amino acid exchanges in positions one and six. J Accid Emerg Med, 1996 Jul, 13(4), 283 - 4 Is it safe to use preprepared endotracheal tubes in the resuscitation room? Bleetman A, Ashwood N. OBJECTIVE: To investigate the bacteriological safety of preprepared endotracheal tubes in accident and emergency departments . METHODS: Swabs were taken and cultured from luminal and exterior surfaces of every preprepared exposed endotracheal tube in a hospital resuscitation room . RESULTS: Coagulase negative staphylococci (common skin commensals) were isolated from 50% of the tubes . No other bacteria were isolated . CONCLUSIONS: The practice of leaving preprepared airway equipment exposed in the resuscitation room is unlikely to contribute to the development of nosocomial pneumonia, provided the equipment is kept dry and that personnel minimize handling. J Clin Microbiol, 1996 Jul, 34(7), 1603 - 5 Evaluation of Vitek GPS-SA card for testing of oxacillin against borderline-susceptible staphylococci that lack mec; Knapp CC et al.; Fifty-one Staphylococcus aureus strains lacking mec for which oxacillin MICs were 1 to 8 micrograms/ml were tested against oxacillin and the combination of oxacillin and clavulanic acid with the Vitek GPS-SA card, the reference broth microdilution method, and the oxacillin agar screen plate . Of the 51 strains, 44 (86%) did not grow on the oxacillin agar screen plate, broth microdilution MICs were 1 to 2 micrograms/ml, and GPS-SA card MICs were < or = 2 micrograms/ml, with the exception of 3 strains that failed to grow in the card on repeated attempts . Another seven strains did grow on the oxacillin agar screen plate . For four of the latter group of strains, oxacillin broth microdilution MICs were > 4 micrograms/ml and GPS-SA card MICs were > or = 4 micrograms/ml; for the other 3 strains, corresponding MICs were 4 and < or = 2 micrograms/ml, respectively . The GPS-SA card classified 86% of strains as oxacillin susceptible. Int J Syst Bacteriol, 1996 Jul, 46(3), 792 - 6 Staphylococcus saprophyticus subsp . bovis subsp . nov., isolated from bovine nostrils; Hajek V et al.; A new coagulase-negative subspecies, Staphylococcus saprophyticus subsp . bovis, is described on the basis of a study of five strains isolated from the anterior nares of cows . This subspecies is differentiated from the other novobiocin-resistant staphylococci by its phenotypic properties, cell wall composition, and levels of genetic relatedness . The type strain of the new subspecies is KV 12 (=CCM 4410). J Pediatr, 1996 Jul, 129(1), 63 - 71 Late-onset sepsis in very low birth weight neonates: a report from the National Institute of Child Health and Human Development Neonatal Research Network; Stoll BJ et al.; OBJECTIVE: Late-onset sepsis (occurring after 3 days of age) is an important problem in very low birth weight (VLBW) infants . To determine the current incidence of late-onset sepsis, risk factors for disease, and the impact of late-onset sepsis on subsequent hospital course, we evaluated a cohort of 7861 VLBW (401 to 1500 gm) neonates admitted to the 12 National Institute of Child Health and Human Development (NICHD) Neonatal Research Network centers during a 32-month period (1991 to 1993) . METHODS: The NICHD Neonatal Research Network maintains a prospectively collected registry of all VLBW neonates cared for at participating centers . Data from this registry were analyzed retrospectively . RESULTS: Of 6911 infants who survived beyond 3 days, 1696 (25%) had one or more episodes of blood culture-proven sepsis . The vast majority of infection (73%) were caused by gram-positive organisms, with coagulase-negative staphylococci accounting for 55% of all infections . Rate of infection was inversely related to birth weight and gestational age . Complications of prematurity associated with an increased rate of infection included intubation, respiratory distress syndrome, prolonged ventilation, bronchopulmonary dysplasia, patent ductus arteriosus, severe intraventricular hemorrhage, and necrotizing enterocolitis . Among infants with bronchopulmonary dysplasia, those with late-onset sepsis had a significantly longer duration of mechanical ventilation (45 vs 33 days; p <0.01) . Late-onset sepsis prolonged hospital stay: the mean number of days in the hospital for VLBW neonates with and without late-onset sepsis was 86 and 61 days, respectively (p <0.001) . Even after adjustment for other complications of prematurity, including intraventricular hemorrhage, necrotizing enterocolitis, and bronchopulmonary dysplasia, infants with late-onset sepsis had a significantly longer hospitalization (p <0.001) . Moreover, neonates in whom late-onset sepsis developed were significantly more likely to die than those who were uninfected (17% vs 7%; p <0.000 1), especially if they were infected with gram-negative organisms (40%) or fungi (28%) . Deaths attributed to infection increased with increasing chronologic age . Whereas only 4% of deaths in the first 3 days of life were attributed to infection, 45% of deaths after 2 weeks were related to infection . CONCLUSIONS: Late-onset sepsis is a frequent and important problem among VLBW preterm infants . Successful strategies to decrease late-onset sepsis should decrease VLBW mortality rates, shorten hospital stay, and reduce costs. Kyobu Geka, 1996 Jul, 49(8 Suppl), 693 - 6 {Successful redo-AVR with cryopreserved allograft for prosthetic valve endocarditis : report of a case}; Imanaka K et al.; A 20-year-old male undergoing mechanical aortic valve replacement suffered from prosthetic valve endocarditis by Coagulase negative Staphylococci 3 months postoperatively . Echocardiography revealed left ventricular distention, severe paravalvular leakage, valve detachment and large periannular abscess adjacent to the left main coronary artery . Redo-AVR was successfully performed with a cryopreserved aortic allograft by cylinder technique . The orifice of abscess was covered with the aortic wall of the graft . Postoperative echocardiography showed trivial central regurgitation and the abscess cavity went on vanishing . He discharged from hospital 2 months later without any symptom. Infect Immun, 1996 Jul, 64(7), 2804 - 7 Role of intercellular adhesion molecule 1 in pathogenesis of staphylococcal arthritis and in host defense against staphylococcal bacteremia; Verdrengh M et al.; Intercellular adhesion molecule 1 (ICAM-1) is a member of the immunoglobulin superfamily that interacts with two integrins, LFA-1 and Mac-1 . These interactions are critical for leukocyte extravasation into inflamed tissue . To assess the role of ICAM-1 expression in the pathogenesis of bacterial infection, homozygously mutant mice lacking the ICAM-1 gene were exposed to Staphylococcus aureus . Within 6 days after inoculation 50% of the animals in the ICAM-1(-/-) group, but none of the controls, had died . Despite the high level of mortality, ICAM-1(-/-) mice developed less frequent and less severe arthritis than their wild-type littermates . In agreement, normal mice inoculated with staphylococci and administered anti-ICAM-1 antibodies exhibited a higher frequency of mortality but less severe arthritis than the controls . Our results indicate that ICAM-1 on the one hand provides protection against systemic disease but on the other hand aggravates the local disease manifestation. Am J Med, 1996 Jul, 101(1), 68 - 76 Oral antibiotic treatment of right-sided staphylococcal endocarditis in injection drug users: prospective randomized comparison with parenteral therapy; Heldman AW et al.; PURPOSE: To compare the efficacy and safety of inpatient oral antibiotic treatment (oral) versus standard parenteral antibiotic treatment (intravenous) for right-sided staphylococcal endocarditis in injection drug users . PATIENTS AND METHODS: In a prospective, randomized, non-blinded trial, febrile injection drug users were assigned to begin oral or intravenous (IV) treatment on admission, before blood culture results were available . Oral therapy consisted of ciprofloxacin and rifampin . Parenteral therapy was oxacillin or vancomycin, plus gentamicin for the first 5 days . Antibiotic dosing was adjusted for renal dysfunction . Administration of other antibacterial drugs was not permitted during the treatment or follow-up periods . Bacteremic subjects having right-sided staphylococcal endocarditis received 28 days of inpatient therapy with the assigned antibiotics . Test-of-cure blood cultures were obtained during inpatient observation 6 and 7 days after the completion of antibiotic therapy, and again at outpatient follow-up 1 month later . Criteria for treatment failure and for drug toxicity were prospectively defined . RESULTS: Of 573 injection drug users who were hospitalized because of a febrile illness and suspected right-sided staphylococcal endocarditis, 93 subjects (16.2%) had two or more sets of blood cultures positive for staphylococci; 85 of these bacteremic subjects (14.8%) satisfied diagnostic criteria for at least possible right-sided staphylococcal endocarditis (no other source of bacteremia was apparent) and entered the trial . Forty-four (oral, 19; IV, 25) of these 85 subjects completed inpatient treatment and evaluation including test-of-cure blood cultures . There were four treatment failures (oral, 1 {5.2%}; IV, 3 {12.0%}; not significant, Fisher's exact test) . Drug toxicity was significantly more common in the parenterally treated group (oral, 3%; IV, 62%; P < 0.0001), consisting largely of oxacillin-associated increases in liver enzymes . CONCLUSIONS: For selected patients with right-sided staphylococcal endocarditis, oral ciprofloxacin plus rifampin is effective and is associated with less drug toxicity than is intravenous therapy. Ann Thorac Surg, 1996 Jul, 62(1), 48 - 53 Long-term results of operation for paravalvular abscess; d'Udekem Y et al.; BACKGROUND . Operation for infective endocarditis with paravalvular abscess is reportedly associated with high mortality and morbidity rates . In an attempt to improve surgical outcome, an approach of radical resection of the abscess and inflamed tissues and reconstruction of the heart with either fresh or glutaraldehyde-fixed bovine pericardium was adopted by two surgeons at our institution . METHODS . From 1979 to 1995, 70 consecutive patients with active infective endocarditis and paravalvular abscess underwent operation . Their mean age was 49 years (range, 16 to 75 years), and 50 patients (71%) were men . Thirty-four patients had native and 36 had prosthetic valve endocarditis (8 had had composite replacement of the aortic valve and ascending aorta) . Most patients (78%) were in New York Heart Association functional class IV . The principal indication for operation was cardiogenic or septic shock in 11 patients, or one or more of the following: persistent sepsis despite adequate antibiotic therapy in 36, congestive heart failure in 31, and recurrent emboli in 16 . Staphylococci were responsible for the infection in 37 patients (53%) . The abscess was in the mitral annulus in 11 patients, in the aortic root in 44, and in the aortic root and at least one other annulus in 15 . After wide resection of the abscess, we reconstructed the heart and annuli with autologous or bovine pericardium . Mechanical heart valves were implanted in 36 patients, bioprostheses in 30, and aortic homografts in 2; valve repair was possible in 2 . Sixteen patients required composite replacement of the ascending aorta and aortic valve . RESULTS . There were 9 operative deaths (13%) . Infections caused by staphylococci and infections in multiple annuli were associated with increased operative mortality rates . Only 1 patient had persistent infection and required reoperation . The mean follow-up was 56 +/- 40 months . There were 12 late deaths, mostly cardiac . The actuarial survival including operative deaths was 64% +/- 8% at 8 years . In 8 patients, recurrent infective endocarditis developed 10 to 102 months after operation . The freedom from recurrent endocarditis was 76% +/- 10% at 8 years . CONCLUSIONS . This experience indicates that radical resection of the abscess and reconstruction of the heart with pericardium yield an excellent chance of eradicating the infection in patients with infective endocarditis and paravalvular abscess . The type of valve implanted may not be as important as radical resection of the abscess . These patients appear to have a greater than average risk of recurrent endocarditis. J Biol Chem, 1996 Jun 28, 271(26), 15803 - 9 Molecular cloning and expression of the gene for elastin-binding protein (ebpS) in Staphylococcus aureus; Park PW et al.; Interactions between staphylococci and components of the extracellular matrix mediate attachment of the bacteria to host tissues and organs and define an important mechanism leading to colonization, invasion, and formation of metastatic abscesses . We have previously demonstrated a specific binding interaction between Staphylococcus aureus and elastin, one of the major protein components of the extracellular matrix . Available evidence suggests that this association is mediated by a 25-kDa elastin-binding protein on the surface of S . aureus (EbpS) . To study the molecular structure and function of EbpS, the gene encoding EbpS was cloned, sequenced, and expressed in Escherichia coli . DNA sequence data indicate that the ebpS open reading frame consists of 606 base pairs and encodes a novel polypeptide with a predicted molecular mass of 23,345 daltons and pI of 4.9 . A polyclonal antibody raised against recombinant EbpS interacted with the native 25-kDa cell surface EbpS and inhibited staphylococcal elastin binding . Furthermore, recombinant EbpS bound specifically to immobilized elastin and inhibited binding of S . aureus to elastin . A degradation product of recombinant EbpS lacking the first 59 amino acids of the molecule and a C-terminal fragment of CNBr-cleaved recombinant EbpS, however, did not interact with elastin . Together, these results confirm that EbpS is the cell surface molecule mediating binding of S . aureus to elastin . The inability of truncated forms of recombinant EbpS to bind to elastin suggests that the elastin binding site in EbpS is contained in the first 59 amino acids of the molecule. J Antimicrob Chemother, 1996 Jun, 37(6), 1103 - 9 Influence of inoculum, medium and serum on the in-vitro susceptibility of coagulase-negative staphylococci to teicoplanin and vancomycin; Kennedy HF et al.; The susceptibility to teicoplanin of 100 coagulase negative staphylococci, predominantly isolated from intravenous catheter tips and exit sites was determined by agar dilution on IsoSensitest media with and without the addition of 20% inactivated horse serum using an inoculum of 10(4) cfu/spot . Incorporation of serum resulted in a three-fold increase in the geometric mean MIC and a four-fold increase in the geometric mean MBC . Further tests, performed in DST and IsoSensitest medium supplemented with 20% inactivated horse serum, resulted in a wide variation in teicoplanin MICs with differences in the geometric mean MICs being up to 8.8-fold in serum media compared to unsupplemented media, whereas the MICs of vancomycin were unaffected . Under the various experimental conditions used, the susceptibility of the coagulase negative staphylococci to teicoplanin varied from 31-100%, while all were consistently susceptible to vancomycin . We therefore recommend that teicoplanin MICs be determined in the presence of serum as these are better related to serum drug levels and reflect more accurately the conditions in vivo . Trials of teicoplanin as monotherapy are required to provide further insight into the relationship between its in-vitro antibacterial activity and its clinical efficacy as the drug is easier to administer, better tolerated and less toxic than vancomycin which might compensate for its reduced activity in the presence of serum. Infect Control Hosp Epidemiol, 1996 Jun, 17(6), 356 - 9 Vancomycin use in a university medical center: comparison with hospital infection control practices advisory committee guidelines; Evans ME et al.; OBJECTIVE: To compare actual vancomycin use in a hospital with the guidelines proposed by the Hospital Infection Control Practices Advisory Committee (HICPAC) of the Centers for Disease Control and Prevention . DESIGN: One-month prospective survey of all patients given vancomycin . Data were collected from patient and laboratory records and discussions with prescribing physicians . SETTING: Four hundred sixty-one-bed tertiary-care university hospital . RESULTS: Only 35% of the 101 vancomycin orders written during the audit were consistent with HICPAC guidelines . Twenty-eight patients were begun on vancomycin for the treatment of documented infections due to beta-lactam-resistant gram-positive bacteria or because preliminary culture results suggested such bacteria were present . Seven methicillin-resistant Staphylococcus aureus, two ampicillin-resistant enterococci, and 10 coagulase-negative staphylococci were recovered . Vancomycin use was judged inappropriate in 66 patients . The drug was used empirically without obtaining appropriate cultures (44 patients), as surgical prophylaxis (6 patients), for primary treatment of antibiotic-associated colitis (3 patients), for convenience in a hemodialysis patient (1 patient), or for other reasons not recommended by HICPAC (12 patients) . CONCLUSIONS: Vancomycin use in our teaching center often was inconsistent with HICPAC guidelines . Information from the audit will be useful for designing strategies to improve vancomycin use. J Heart Lung Transplant, 1996 Jun, 15(6), 646 - 9 Infective endocarditis of the tricuspid valve in an orthotopic heart transplant recipient; Stewart MJ et al.; A 43-year-old orthotopic heart transplant recipient had coagulase-negative staphylococcus endocarditis 26 weeks after the operation . A diagnosis of endocarditis was confirmed and followed up by serial transoesophageal echocardiography . Treatment with intravenous gentamycin and vancomycin cured her endocarditis, and a 2.5 cm vegetation regressed significantly . She has been well since and, at 14 months after transplantation, was back to her normal activities . Although repeated blood culture yielded only intermittent light growths of coagulase-negative staphylococci, there were several positive samples . In a setting of infective features, light growths of coagulase-negative staphylococcus should be taken seriously if repeatedly positive in heart transplant recipients or other immunocompromised patients . Transesophageal echocardiography offers significant advantages over the transthoracic modality in suspected endocarditis. Burns, 1996 Jun, 22(4), 279 - 82 Staphylococcal infections in the Sofia Burn Centre, Bulgaria; Lesseva MI et al.; This study analysed staphylococcal infections in the Sofia Burns Centre in order to estimate their frequency, features and role in burns . For an 8-year period (1987-94) the bacterial aetiology of wound infections and bacteraemia in burned patients was studied . The prevalence of staphylococci in both wound exudation (31.4 per cent) and in blood cultures (60.7 per cent) was established . During the last year of the study there was a significant increase in the incidence of methicillin-resistant Staphylococcus aureus (MRSA) from 19.4 per cent in 1993 to 28.0 per cent in 1994 (P < 0.001) . This raised serious therapeutic problems . MRSA were found more frequently in the ICU than in the wards and in wounds and blood cultures compared with other species/strains . MRSA caused infections in 18.8 percent of patients . Over 70 per cent of the MRSA strains were resistant to gentamicin, erythromycin and tetracycline and about one-third to lincomycin, co-trimoxazole, chloramphenicol and ciprofloxacin . All the MRSA strains were sensitive to vancomycin and 71.1 per cent to rifampicin . These findings show the necessity of urgent measures for restriction of the further distribution of MRSA infections in our burns centre. FEMS Microbiol Lett, 1996 Jun 1, 139(2-3), 109 - 15 Immunological crossreactivity to the catabolite control protein CcpA Bacillus megaterium is found in many gram-positive bacteria; Kuster E et al.; The catabolite control protein CcpA from Bacillus megaterium was overproduced as a fusion protein to a 6xhis affinity tag and purified to homogeneity . Polyclonal antibodies of high affinity and specificity were raised against the purified protein . The serum did not crossreact with purified Lac repressor despite the fact that CcpA and LacI belong to the same protein family . Using this antiserum we identified proteins that share antigenic determinants with CcpA in many Gram-positive bacteria, including bacilli, staphylococci, lactic acid bacteria, and some actinomycetes. J Neurol Neurosurg Psychiatry, 1996 Jun, 60(6), 671 - 5 Reduced bacterial adhesion to hydrocephalus shunt catheters mediated by cerebrospinal fluid proteins; Brydon HL et al.; BACKGROUND--Prosthetic infections are a major problem, requiring complex and lengthy management . The role of blood proteins in the pathogenesis of implant infection has been investigated, but research into the role of CSF protein in shunt infections has not been undertaken, even though a high CSF protein has been assumed to increase the risk of such infections . METHODS--New shunt catheters were exposed to either CSF or individual protein solutions, and the numbers of radiolabelled staphylococci that adhered to them were compared with controls that had been exposed to saline only . RESULTS--A significant reduction in bacteria adhering to the test catheter was found in each instance . Furthermore, the CSF with the highest protein content, from a patient with intraventricular haemorrhage, had the greatest inhibitory effect on bacterial adhesion . The effect of the solutions on the hydrophobicity of the silicone rubber was also investigated . The silicone rubber was more hydrophilic, and bacterial adhesion was less, with solutions containing a higher protein content, and these findings were in keeping with the current theories on the mechanism of bacterial adhesion to polymers . CONCLUSIONS--A high CSF protein content does not predispose to the development of shunt infections. Arch Intern Med, 1996 May 27, 156(10), 1109 - 12 A 1-year trial of nasal mupirocin in the prevention of recurrent staphylococcal nasal colonization and skin infection; Raz R et al.; BACKGROUND: The usefulness of nasal mupirocin in preventing recurrent staphylococcal nasal colonization and skin infection has been examined in immunodeficient patients and in healthy staphylococcal carriers but not in immunocompetent staphylococcal carriers who experience recurrent skin infections . We studied 34 such patients . METHODS: After an initial 5-day course of nasal mupirocin ointment for all patients, 17 patients continued to apply a 5-day course of nasal mupirocin every month for 1 year, and the other 17 patients applied a placebo ointment . Nasal cultures were obtained monthly, and all episodes of skin infection were recorded . RESULTS: The overall number of positive nasal cultures was 22 in the mupirocin group and 83 in the placebo group (P < .001), and the number of skin infections was 26 and 62, respectively (P < .002) . Eight of the 17 mupirocin-treated patients but only 2 in the placebo group remained free of positive staphylococcal nasal cultures . One of the 10 patients who were free of colonization during the 12-month treatment period had skin infections, in contrast to all 24 of the patients with positive cultures (P < .01) . Staphylococci resistant to mupirocin were observed in 1 patient . No adverse effects were reported . CONCLUSIONS: A monthly application of mupirocin ointment in staphylococcal carriers reduces the incidence of nasal colonization, which in turn lowers the risk of skin infection. J Card Surg, 1996 May-Jun, 11(3), 180 - 6 Surgical management of infected permanent transvenous pacemaker systems: ten year experience; Vogt PR et al.; BACKGROUND: Between January 1985 and June 1995, more than 1800 consecutive patients underwent implantation of a new permanent cardiac pacemaker at our institution . Thirty-six patients (0.02%) had 45 reinterventions for infected pacemaker systems . METHODS: in group A, 24 of 27 patients received simultaneous implantation of a new pacemaker . One had reimplantation of the same pacemaker in the same pocket, and two did not require reimplantation . The leads were retained in 19 (70%) of the patients . In group B, nine patients underwent cardiopulmonary bypass or "pursestring" surgery for removal of an infected pacemaker; a new epicardial pacemaker system was simultaneously implanted in seven patients . RESULTS: Identification of an infectious agent failed in 17 patients (47%), and Staphylococci were found in 15 patients (42%) . The time from pacemaker implantation to onset of infection ranged from 1 month to 11 years (mean 31 months; median 19 months) and the time from onset of infection to surgical treatment from 1 month to 7 years (mean 7 months; median 2 months) . The mean follow-up time is 74 months (range, 1 month to 10 years; median 5 years) . There were 9 reoperations in 3 patients (16%) of group A for recurrent infection of their retained leads ultimately necessitating the use of open cardiac surgery . There was no early death; six patients died late due to unrelated causes . CONCLUSIONS: Complete removal of all pacemaker leads is recommended; open heart surgery with the use of cardiopulmonary bypass is indicated in selected cases and is effective and safe. Diagn Microbiol Infect Dis, 1996 May, 25(1), 9 - 13 Disk diffusion interpretive criteria for fusidic acid susceptibility testing of staphylococci by the National Committee for Clinical Laboratory Standards method; Coutant C et al.; Fusidic acid is used in many countries for the treatment of multiresistant staphylococcal infection, especially multiresistant Staphylococcus aureus infection (MRSA) . We collected consecutive fusidic acid-resistant isolates of staphylococci from the routine laboratory over several years, and compared these strains with fusidic acid-susceptible staphylococci to establish interpretive criteria for disk diffusion testing by National Committee for Clinical Laboratory Standards (NCCLS) methods . The minimum inhibitory concentrations (MICs) and zone diameters for strains of S . aureus (n = 102), including MRSA, S . saprophyticus (n = 20) and other coagulase-negative staphylococci (n = 115) were determined by NCCLS agar dilution and disk diffusion tests using a 2.5-micrograms disk of fusidic acid . MICs were bimodally distributed . No isolates had MICs of 0.5 or 1 microgram/ml; thus, we chose these values to define strains of intermediate susceptibility . The error-rate-bounded method was used to determine interpretive zone diameters for disk testing . Interpretive zone diameter criteria were found to be: susceptible > or = 22 mm, intermediate 18-21 mm, and resistant < or = 17 mm . All S . saprophyticus were intrinsically resistant to fusidic acid (MIC > or = 2 micrograms/ml). Eur J Clin Microbiol Infect Dis, 1996 May, 15(5), 405 - 7 Staphylococcus lugdunensis as a cause of abscesses in the perineal area; Ortiz de la Tabla V et al.; The first two patients with skin infections in the perineal area due to Staphylococcus lugdunensis are described . One had an abscess of the Bartholin gland, and the other presented with several cutaneous abscesses in the pubic area, which had a prolonged and recurrent clinical course despite appropriate antibiotic treatment . This report emphasizes the pathogenic role of Staphylococcus lugdunensis and the importance of identifying coagulase-negative staphylococci to species level in some instances. Ann Vasc Surg, 1996 May, 10(3), 244 - 53 Local application of vancomycin for prophylaxis of graft infection: release of vancomycin from antibiotic-bonded Dacron grafts, toxicity in endothelial cell culture, and efficacy against graft infection in an animal model; Muhl E et al.; Methicillin-resistant strains of Staphylococcus epidermidis cause an increasing number of prosthetic infections . This prompted us to test the uptake of vancomycin in various graft materials in vitro, its influence on graft healing, and its efficacy against graft infection in pigs . Incubation of six different Dacron graft materials in a vancomycin solution (20 gm/L) was performed . Grafts were then placed in plasma, and samples were taken over 72 hours to determine vancomycin levels . Release of vancomycin ranged from 775 micrograms/cm2 to 3691 micrograms/cm2 after 1 hour of incubation . Gelatin-covered grafts increased release of vancomycin fourfold when incubation time was extended to 24 hours: uncovered grafts or the collagen-covered graft did not . Graft healing was not complicated when a vancomycin-bonded, gelatin-impregnated Dacron graft was implanted to replace the common femoral artery in pigs . Four weeks after implantation, histologic examination revealed normal development of neointima and perigraft scar tissue in the vancomycin-treated (n = 4) and untreated (n = 5) grafts . To test the efficacy of local vancomycin against graft infection, grafts were implanted in the groin of pigs and contaminated with 2 x 10(7) colony-forming units of Staphylococcus aureus . Four weeks after implantation, all grafts were infected in the untreated group (n = 6), with abscess, nonincorporated graft, and detection of S . aureus from the graft . In the treatment group (n = 6) vancomycin was added to the contaminated grafts . As a carrier for the vancomycin, we used a resorbable gelatin-glycerol foam . All grafts healed without infection . The difference between the treated and untreated groups is statistically significant (p < 0.05) . We conclude that it may be effective to prevent graft infection with local application of vancomycin if an in situ replacement of infected graft (infected by gram-positive bacteria) is necessary or if there is a high risk of infection by methicillin-resistant- staphylococci. Zh Mikrobiol Epidemiol Immunobiol, 1996 May-Jun, (3), 78 - 80 {The dynamics of the persistence characteristics of staphylococci under the action of the microclimate of a speleotherapy mine}; Chernova OP et al.; A decrease in the persistence characteristics of staphylococci under the influence of microclimate in a spelean pit has been demonstrated under experimental conditions . Clinical investigations have confirmed the capacity of speleotherapy to decrease the microbial contamination of the upper respiratory tract and to inhibit the persistence properties of staphylococcal microflora in children with respiratory allergosis, which seems to be the basis of the positive effect achieved by treatment with microclimate in a spelean pit. Zh Mikrobiol Epidemiol Immunobiol, 1996 May-Jun, (3), 68 - 70 {The construction of a diagnostic model for the differentiation of resident and transient staphylococcal microflora in bacterial carriage}; Bukharin OV et al.; The comparative analysis of 16 biological characteristics of staphylococci isolated from specially selected groups of persons with different types of carrier state has been made . Five signs (antilysozyme, anti-interferon and anticomplementary activities, sensitivity to fusidin, microbial contamination) forming the main informative combination have been determined . The diagnostic model of the differentiation of resident and transitory staphylococcal microflora in carrier state has been developed. J Antimicrob Chemother, 1996 May, 37(5), 901 - 9 Choice of a routine method for detecting methicillin-resistance in staphylococci; Wallet F et al.; Four methods were compared for their abilities to detect methicillin-resistance of Staphylococcus strains using mecA gene PCR analysis in 6 h as the gold standard . 57 Staphylococcus aureus and 100 coagulase negative staphylococci (CNS) were evaluated by the oxacillin disc diffusion method (Kirby-Bauer), the automated API (ATB-plus) system (bioMerieux, La Balme les Grottes, France) in 24 h, the rapid BBL Crystal MRSA ID system (Becton Dickinson, Cockeysville, Md.), the oxillin MICs using the NCCLS agar dilution method in 24 h, and the mecA gene PCR analysis . For S . aureus, the correlation was excellent between the BBL Crystal MRSA ID system and mecA gene PCR analysis (positive predictive value = 100%; negative predictive value = 97%) and oxacillin MIC (positive predictive value = 96%; negative predictive value = 96%) . The correlation between BBL Crystal MRSA ID and mecA gene PCR was not reliable for CNS (negative predictive value = 68%) . For CNS, the slower routine susceptibility methods to identify intrinsic methicillin-resistance were better: API ATB Staph has a positive predictive value = 94% and a negative predictive value = 82%, and the disc diffusion test has a positive predictive value = 95% and a negative predictive value = 74% . However, BBL Crystal MRSA ID was as reliable as some of the other methods tested for CNS after 6 h incubation when the inoculum was increased: positive predictive value = 94% and a negative predictive value = 77% . These results emphasize that genotypic detection of methicillin-resistance will undoubtedly become important to detect rapidly methicillin-resistance, especially for CNS. Br J Dermatol, 1996 May, 134(5), 824 - 30 Ultraviolet B-exposed major histocompatibility complex class ii positive keratinocytes and antigen-presenting cells demonstrate a differential capacity to activate T cells in the presence of staphylococcal superantigens; Skov L et al.; In this study we tested the capacity of ultraviolet B (UVB)-irradiated major histocompatibility complex (MHC) class II+ keratinocytes, monocytes and dendritic cells to activate T cells in the presence of Staphylococcus enterotoxin B . We demonstrated that UVB irradiation of MHC class II+ keratinocytes does not change their capacity to activate T cells in the presence of Staphylococcus enterotoxin B . In contrast, UVB irradiation of antigen-presenting cells decreases their capacity to activate T cells . This differential capacity to activate T cells after UVB irradiation was not due to factors released from UVB-irradiated cells . The interferon-gamma induced upregulation of HLA-DR and intercellular adhesion molecule-1 on keratinocytes does not seem to be the only explanation, since UVB irradiation decreased the accessory cell function of interferon-gamma pretreated monocytes . Differential requirements for and UVB regulation of costimulatory molecules may be involved, since blocking of the B7/CD28 pathway affects the capacity of dendritic cells but not keratinocytes to activate T cells in the presence of Staphylococcus enterotoxin B . Thus, MHC class II+ keratinocytes in the presence of superantigens released from staphylococci may activate T cells and maintain inflammation despite UVB treatment. Infect Control Hosp Epidemiol, 1996 May, 17(5), 276 - 80 Pediatric primary gram-negative nosocomial bacteremia: a possible relationship with infusate contamination; Macias-Hernandez AE et al.; OBJECTIVE: To evaluate the potential contribution of "extrinsic" contamination of intravenous fluids in hospital bacteremia and infection . DESIGN: Prospective cross-sectional survey of infusate contamination, December 1992 to December 1993 . SETTING: A pediatric department (1,500 admissions per year) in a general, urban teaching hospital, serving low-income patients . SAMPLES AND PATIENTS: Infusate samples (0.5 to 1.0 mL) from the injection port used by the staff were taken for cultures from all febrile or septic patients in hospital wards . At least four samples were taken each day; if no febrile or septic patients were available, other patients were sampled at convenience . RESULTS: A 6.8% positive culture rate (87 contaminates in 1,277 infusates) was obtained, without significant differences among the wards . Gram-negative organisms were recovered from 56 samples (62.9%), mainly of the tribe Klebsielleae (56.1%) . Coagulase-negative staphylococci were isolated in 30 samples (33.7%) . There was no significant difference between the febrile-septic group and the asymptomatic group in the rate of infusate contamination (P = .59) . In eight patients, the same organisms were recovered from infusate and blood culture . The overall bacteremia rate was 2.5 per 100 discharges . CONCLUSIONS: Compared to previous reports, higher infusate contamination rates and different organisms (mainly gram-negative) were observed . In hospitals of underdeveloped countries, nosocomial infection control frequently is disregarded . Infusate contamination may be common and could lead to gram-negative bacteremia . In such settings, it seems advisable to perform surveillance studies to identify infusate contamination, because a single infusate contamination could be a signal for an epidemic. J Clin Pathol, 1996 May, 49(5), 365 - 8 Significance of endotoxin in lethal synergy between bacteria associated with sudden infant death syndrome: follow up study; Sayers NM et al.; AIM: To investigate the role of endotoxin in synergy between bacterial toxins associated with sudden infant death syndrome (SIDS) . METHODS: Extracellular toxins of 13 isolates of Staphylococcus from SIDS victims and matched healthy infants were tested for lethal toxicity in chick embryos with and without standard endotoxin (used at 1.00 ng/embryo) . Endotoxin and toxins from staphylococci were used at dilutions with negligible lethality . RESULTS: Simultaneous injection of non-lethal levels of endotoxin and toxins from 11 of the 13 staphylococcal isolates tested produced lethal toxicity that was 111 to 613% greater than expected by an additive effect alone . This was highly significant and occurred even in the absence of staphylococcal enterotoxins or toxic shock syndrome toxin-1 . CONCLUSION: Endotoxin enhancement of staphylococcal toxin lethality could be partly responsible for the clinical outcome in SIDS. J Invest Dermatol, 1996 May, 106(5), 982 - 8 Cutaneous exposure to the superantigen staphylococcal enterotoxin B elicits a T-cell-dependent inflammatory response; Saloga J et al.; We analyzed the impact of superantigens secreted by skin-colonizing Staphylococci on the skin and the associated lymphoid tissue following epicutaneous application and intracutaneous injection of small amounts of staphylococcal enterotoxin B (SEB) . A single intracutaneous injection of 50 ng of SEB elicited a strong inflammatory response in the skin of BALB/c mice . Three to 6 h later, we observed langerhans cell activation, mast cell degranulation, vasodilation, upregulation of ICAM-1, and induction of VCAM-1 on dermal blood vessels, with vascular adhesion of granulocytes . by 12 to 24 h, cell infiltration of the dermis increased, reaching the epidermis . Among the infiltrating leukocytes, a substantial number of eosinophils was found . After 48 h, the infiltrate was dominated by mononuclear cells . The response to SEB was dose-dependent, and signs of inflammation slowly disappeared over 5 to 7 days . Although the induction of VCAM-1 on dermal blood vessels suggested a role for interleukin-1/tumor necrosis factor-alpha in this reaction, the activation of monocytes/macrophages was not able to substitute for lymphocytes, as severe combined immunodeficiency (SCID) mice (which are lymphocyte-deficient) did not mount an inflammatory skin response to intradermal injection of SEB . The fact that nude mice (T-cell-deficient) also did not mount an inflammatory response to SEB indicated the T-cell dependency of the response . The V beta specificity of the SEB effect was demonstrated by the fact that SJL/J mice, which lack V beta 8+ T cells (the major SEB-reactive T cell population in mice), exhibited much weaker responses . Deletion or tolerization of SEB-reactive V beta T cells was not observed after a single intradermal injection of such minute amounts of SEB. Toxicology, 1996 Apr 30, 108(3), 207 - 15 Species specificity of 2-aryl carbapenem-induced immune-mediated hemolytic anemia in primates; Lankas GR et al.; L-695,256 is a novel 2-fluorenonyl carbapenem antibiotic with significant antimicrobial activity against strains of methicillin-resistant Staphylococci . This prototype compound was administered intravenously to rhesus monkeys (Macaca mulatta) at does of 50 or 200 mg/kg/day for 2 weeks to assess toxicity and found to induce a hemolytic anemia characterized by extravascular hemolysis and splenomegaly . A subsequent study in this species in which 100 mg/kg/day was administered i.v . for 4 weeks showed that all animals were direct antiglobulin test (Coombs' test) positive for IgG with 20-25% reductions in the erythron . Following 3 weeks of recovery, the erythron had returned to normal, although it took an additional 2 months for the Coombs' test to become negative . Challenge of these same animals with 0.5 million U/kg (300 mg/kg/day) of penicillin intravenously indicated no apparent cross-reactivity . Since attempts to establish a model for this immune-mediated hemolytic anemia with this drug in rats or mice were unsuccessful, a 2-week i.v . study in squirrel monkeys (Saimiri sciureus) was conducted at a dose of 200 mg/kg/day . All animals in this study remained Coombs' test negative with no changes in the erythron, suggesting an increased sensitivity to beta-lactam-induced anemia in rhesus monkeys compared to other species . Further support for this hypothesis was obtained using the cephalosporin antibiotic, cefotetan . This compound induced a high incidence of Coombs' test positive hemolytic anemia at clinically relevant doses in rhesus monkeys, despite a very low incidence of this adverse effect in patients with many years of clinical use . These data suggest that although rhesus monkeys respond in a qualitatively similar manner to humans with regard to high doses of beta-lactam antibiotics, their sensitivity may overestimate the risk of immune-mediated hemolytic anemia for clinical use. Tidsskr Nor Laegeforen, 1996 Apr 30, 116(11), 1328 - 30 {Bacterial endocarditis in premature children . A complication caused by central venous catheters}; Brunvand L et al.; During the last ten years, the literature has included an increasing number of reports of bacterial endocarditis in prematurely born neonates . We describe the cases of two prematurely born infants with structurally normal hearts who, when examined by echo cardiography, were shown to have intercardial vegetations . They were diagnosed as having infective endocarditis caused by coagulase negative staphylococci . Both infants had central venous lines and received total parenteral nutrition . Both infants were treated successfully with antibiotics . One of them died later of sudden infant death syndrome. Ugeskr Laeger, 1996 Apr 29, 158(18), 2538 - 41 {Peritonitis in continuous ambulatory peritoneal dialysis . Culture of peritoneal dialysate fluid}; Moller JK et al.; Conventional aerobic and anaerobic culture of peritoneal dialysate effluent from patients in continuous peritoneal dialysis (CAPD) was compared to culture in a semiautomated blood culture system . During a two-year period 78 of 79 consecutive episodes of peritonitis among 45 Danish CAPD patients were cultured and the etiology of the infection found in 73 (94%) . The sensitivity of the blood culture system was 88%, whereas the sensitivity of the conventional culture of the dialysate effluent was 81% . This difference is not significant (McNemar test; 0.5 > p > 0.3) . The majority of isolates were Gram-positive bacteria dominated by coagulase-negative staphylococci (38%) . In comparison, only 2% of the cultures of peritoneal dialysate effluent taken within the same period from patients without clinical signs of peritonitis were positive . All the Gram-positive aerobic bacteria were sensitive to vancomycin whereas 97% of the Gram-negative aerobic bacteria were sensitive to gentamicin . An initial empiric treatment of peritonitis with a combination of vancomycin and gentamicin is recommended. Ugeskr Laeger, 1996 Apr 29, 158(18), 2532 - 7 {Peritonitis in continuous ambulatory peritoneal dialysis . An evaluation of the empiric initial antibiotic treatment}; Hagelskjaer LH et al.; Retrospectively, the clinical outcome and the initial empiric antibiotic treatment of peritonitis in 106 patients on continuous ambulatory peritoneal dialysis (CAPD) were evaluated during a two-year period . A mean frequency of 0.89 episodes of peritonitis per year of dialysis was found . There was a tendency towards an increased frequency of peritonitis in older patients . Diabetic patients constituted a younger age group and had a tendency towards having a lower risk of peritonitis . Patients with polycystic renal disease had a significantly increased risk . The risk of episodes with coagulase-negative staphylococci increased significantly with age . Repeated peritonitis episodes with coagulase-negative staphylococci was associated with a significant increase in the appearance of methicillin drug resistance . Carriers of Staphylococcus aureus had a significantly increased risk of Staphylococcus aureus peritonitis . Microorganisms were cultured in 94% of the episodes . The initial antibiotic therapy was only sufficient in 66% due to antimicrobial drug resistance . The initial antibiotic treatment was changed in 58% of the episodes . The treatment could have been changed to antibiotics with a narrower antimicrobial spectrum in 51% of the episodes . Relapse was seen in 11% of culture positive episodes . In 16% of the episodes (29% of patients with peritonitis) it was necessary to remove the dialysis catheter and transfer the patient to haemodialysis to clear the infection . Only 15% of these patients returned to CAPD again . We found that an initial empiric antibiotic regime of vancomycin combined with an aminoglycoside is to be recommended as achieving an antibiotic coverage of 88%, and this is now the standard regime in the department. J Med Chem, 1996 Apr 26, 39(9), 1864 - 71 Synthesis and structure-activity relationship of (lactamylvinyl)cephalosporins exhibiting activity against staphylococci, pneumococci, and enterococci; Heinze-Krauss I et al.; The synthesis and structure-activity relationships of a new class of vinylcephalosporins substituted with a lactamyl residue are described . These compounds show excellent activity against enterococci and retain the broad spectrum activity of third-generation cephalosporins such as ceftriaxone. Microb Drug Resist, 1996 Spring, 2(1), 29 - 41 Staphylococcal peptidoglycan interpeptide bridge biosynthesis: a novel antistaphylococcal target? Kopp U, Roos M, Wecke J, Labischinski H. In staphylococci, crosslinking of the peptide moiety of peptidoglycan is mediated via an additional spacer, the interpeptide bridge, consisting of five glycine residues . The femAB operon, coding for two approximately 50-kDa proteins is known to be involved in pentaglycine bridge formation . Using chemical mutagenesis of the beta-lactam-resistant strain BB270 and genetic, biochemical, and biophysical characterization of mutants selected for loss of beta-lactam resistance and reduced lysostaphin sensitivity it is shown that peptide bridge formation proceeds via three intermediate bridge lengths (cell wall peptides with no, one, three, and five glycine units) . To proceed from one intermediate to the next, three genes appear necessary: femX, femA, and femB . The drastic loss of beta-lactam resistance after inactivation of FemA or partial impairment of FemX even beyond the level of the sensitive wild-type strains renders these proteins attractive antistaphylococcal targets. J Paediatr Child Health, 1996 Apr, 32(2), 158 - 61 Late-onset infections of infants in neonatal units; Isaacs D et al.; OBJECTIVE: To examine regional variations in the incidence of late-onset neonatal infections in Australian and New Zealand neonatal units . METHODOLOGY: A longitudinal, prospective surveillance study of systemic sepsis (septicaemia or meningitis) in 11 neonatal units: 10 in the Australian States of the Northern Territory, New South Wales, Queensland, Victoria and Western Australia, and 1 in Christchurch, New Zealand . The results are reported of late-onset neonatal infection (defined as sepsis after 48 h) for the second year of prospective surveillance, data being collected from 1 October 1992 to 30 September 1993 . RESULTS: Data were available on 24535 live births in Australia, representing approximately 10% of all live births in the country . There were 320 episodes of sepsis in Australian units affecting 294 babies . One hundred of these episodes (31%) were early-onset; 3.0% of babies admitted to six tertiary care neonatal units attached to maternity hospitals developed late sepsis, and this rate did not differ between units . The proportion of babies infected was inversely related to birthweight: 22.6% of babies under 1OOOg, but 0.6% over 2000g . Coagulase negative staphylococci were the commonest cause of late-onset sepsis . There were 26 episodes of S . aureus septicaemia, of which only one was due to MRSA . Meningitis occurred in 13 babies (5.9%) with late-onset sepsis . The mortality from late-onset sepsis was 7.7% . CONCLUSIONS: Coagulase-negative staphylococci are the commonest cause of late-onset sepsis of babies in neonatal units . There were no major regional differences in the incidence of, or the organisms causing, late sepsis. Pediatr Pol, 1996 Apr, 71(4), 307 - 11 {Staphylococci and enteric rods in the oral cavity of children with acute lymphoblastic leukemia}; Krzeminski Z et al.; The frequency of isolation and the number of staphylococci and Gram-negative enteric rods in saliva were estimated in a group of 48 children of both sexes, aged 4-15 who were in the course of treatment of acute lymphoblastic leukemia (ALL) . The results were compared with the same parameters in a group of 44 healthy children of the same age . Staphylococci (both coagulase-negative and coagulase-positive) were found in the oral cavities of all healthy children and of 91.6% children with ALL . However, the diseased children harbouring staphylococci revealed a significantly higher average number of these bacteria than healthy children; the respective values were 3.59 and 3.02 log CFU/ml of saliva (p = 0.0148) . The average number of staphylococci in saliva was in both groups of children in negative correlation with the number of polymorphonuclear granulocytes in blood . Gram-negative enteric rods were present in the oral cavities of 13.6% of healthy children and 33.3% of children with ALL (p = 0.0005), but the counts of these bacteria were comparable . The negative correlation between the number of these bacteria and the number of polymorphonuclear granulocytes in blood was not significant. Curr Microbiol, 1996 Apr, 32(4), 201 - 7 Staphylococci bind heparin-binding host growth factors; Pascu C et al.; Staphylococcus aureus, which mediated binding to heparan sulfate, and also strains of coagulase-negative staphylococci (CNS) adhered in high numbers to polymers with end-point attached heparin . A characteristic feature of several cell growth factors is strong affinity for heparin . In the present study, binding of the 125I-labeled heparin-binding growth factors (HBGF), acidic and basic fibroblast growth factor (aFGF, bFGF), and platelet-derived growth factor (PDGF) by S . aureus and CNS strains was examined . Staphylococcal strains used in this study bind bFGF and PDGF, but not aFGF . The binding of bFGF and PDGF was time dependent, influenced by pH and ionic strength for S . aureus Cowan 1 . Preincubation of staphylococcal cells with unlabeled bFGF enhanced bFGF binding, but heparin, protamine sulfate, poly-L-lysine, and suramin were potent inhibitors of 125I-bFGF binding to cells of S . aureus Cowan 1 . Glycosaminoglycans of comparable size (chondroitin sulfate), other polysulfated polymers (lambda-carrageenan, fucoidan), and some polysulfated polysaccharides (dextran sulfate, pentosan polysulfate) inhibited binding of both GFs to various extents . The partial inhibition of binding of both GFs after protease and periodate treatments indicates that both proteinaceous and other carbohydrate moieties participate in the binding . A lysozyme cell surface extract and bacterial lysates of S . aureus Cowan 1 competitively inhibited binding of 125I-bFGF and 125I-PDGF . These results suggest that staphylococci have the ability to bind two of the HBGFs, bFGF and PDGF, but not aFGF, via more than one cell structure . These binding structures seem to be exposed on the cell surface and deeply anchored in the cytoplasmic membrane as well. J Clin Microbiol, 1996 Apr, 34(4), 818 - 23 HSP60 gene sequences as universal targets for microbial species identification: studies with coagulase-negative staphylococci; Goh SH et al.; A set of universal degenerate primers which amplified, by PCR, a 600-bp oligomer encoding a portion of the 60-kDa heat shock protein (HSP60) of both Staphylococcus aureus and Staphylococcus epidermidis were developed . However, when used as a DNA probe, the 600-bp PCR product generated from S . epidermidis failed to cross-hybridize under high-stringency conditions with the genomic DNA of S . aureus and vice versa . To investigate whether species-specific sequences might exist within the highly conserved HSP60 genes among different staphylococci, digoxigenin-labelled HSP60 probes generated by the degenerate HSP60 primers were prepared from the six most commonly isolated Staphylococcus species (S . aureus 8325-4, S . epidermidis 9759, S . haemolyticus ATCC 29970, S . schleiferi ATCC 43808, S . saprophyticus KL122, and S . lugdunensis CRSN 850412) . These probes were used for dot blot hybridization with genomic DNA of 58 reference and clinical isolates of Staphylococcus and non-Staphylococcus species . These six Staphylococcus species HSP60 probes correctly identified the entire set of staphylococcal isolates . The species specificity of these HSP60 probes was further demonstrated by dot blot hybridization with PCR-amplified DNA from mixed cultures of different Staphylococcus species and by the partial DNA sequences of these probes . In addition, sequence homology searches of the NCBI BLAST databases with these partial HSP60 DNA sequences yielded the highest matching scores for both S . epidermidis and S . aureus with the corresponding species-specified probes . Finally, the HSP60 degenerate primers were shown to amplify an anticipated 600-bp PCR product from all 29 Staphylococcus species and from all but 2 of 30 other microbial species, including various gram-positive and gram-negative bacteria, mycobacteria, and fungi . These preliminary data suggest the presence of species-specific sequence variation within the highly conserved HSP60 genes of staphylococci . Further work is required to determine whether these degenerate HSP60 primers may be exploited for species-specific microbic identification and phylogenetic investigation of staphylococci and perhaps other microorganisms in general. Zentralbl Bakteriol, 1996 Apr, 283(4), 497 - 501 In-vitro activity of RP 59500, a new semisynthetic injectable pristinamycin against staphylococci; von Eiff C et al.; We compared the in vitro activity of RP 59500, a new streptogramin, with that of vancomycin and ciprofloxacin against 130 strains of S . aureus and 117 strains of coagulase-negative staphylococci, using the agar dilution method . The antistaphylococcal activity of RP 59500 was similar to that of vancomycin . All staphylococcal strains were inhibited by < or = 2 micrograms/mL, including 61 methicillin-resistant S . aureus, 28 methicillin-resistant S . epidermidis and 23 methicillin-resistant S . haemolyticus strains . In contrast to ciprofloxacin, the in vitro activity of RP 59500 as well as of vancomycin remained almost unchanged, irrespective of the resistance phenotype for methicillin. Ophthalmologe, 1996 Apr, 93(2), 126 - 9 {Contamination of intraocular fluid in pars plana vitrectomy}; Egger SF et al.; Endophthalmitis after pars plana vitrectomy is rare, with an incidence of 0.05-0.14% . The aim of this study was to evaluate the microbiological situation during pars plana vitrectomy and to ascertain what organisms and how many enter the eye during the operation . PATIENTS AND METHODS: Twenty-five consecutive subjects undergoing primary pars plana vitrectomy were included in the study . Patients were excluded if they had evidence of local or systemic infections or had undergone antibiotic therapy within 3 weeks before surgery . A standard three-port pars plana vitrectomy was performed on each patient . Preoperative smears of the conjunctiva and intraoperative aspirates of the vitreous were taken immediately after sclerotomy, and aspirates of the intraocular fluid at the conclusion of operation . RESULTS: We obtained preoperative smears from the conjunctival sac of all patients, and found that 19 patients (76%) had positive cultures, with coagulase-negative staphylococci as the most commonly isolated organisms, (n = 14; 56%) . Vitreous--aspirated immediately after sclerotomy--was sterile in 68% (n = 17) . In 32% (n = 8) contamination occurred, the microorganisms isolated being coagulase-negative staphylococci (20%) and Staphylococcus aureus (12%) . Five of the samples (20%) of intraocular fluid from the vitreous cavity--aspirated before wound closure--were contaminated, coagulase-negative staphylococci (8%) and Staphylococcus aureus (12%) again being found in culture . In no case did postoperative endophthalmitis develop . CONCLUSIONS: This study demonstrates that bacteria enter the eye during pars plana vitrectomy and that there is a change in the contaminating bacterial species during operation . Even if bacteria remain in the eye after pars plana vitrectomy, postoperative endopthalamitis does not necessarily develop. J Neurosurg, 1996 Apr, 84(4), 617 - 23 The significance of bacteriologically positive ventriculoperitoneal shunt components in the absence of other signs of shunt infection; Steinbok P et al.; The purpose of this study was to determine the significance of "asymptomatic bacteriological shunt contamination" (ABSC), defined as a positive bacteriological culture found on a ventricular shunt component in the absence of bacteria in the cerebrospinal fluid (CSF) culture and/or clinical evidence of infection . Of 174 ventriculoperitoneal shunt revisions, 19 cases of ABSC were identified and reviewed retrospectively . In all but one case, no antibiotic medications were instituted because of the positive bacteriological culture . The most common infecting organisms were coagulase-negative staphylococci (seven) and propionibacteria (eight) . A comparison of the 19 study cases with the authors' overall shunt experience, as documented in the British Columbia's Children's Hospital shunt database for the time period of the study, lead the authors to suggest that ABSC was not of significance in causing the shunt failure at which contamination was identified and, more importantly, did not increase the risk of future shunt malfunction . The results of this study indicate that in the absence of clinical evidence of shunt infection or a positive bacteriological culture from CSF, bacteria in a shunt component removed at revision in a child almost always represents a contaminant that may be ignored . Therefore, the authors advise that routine culture of shunt components removed at revision of a shunt is not indicated. Am J Clin Pathol, 1996 Apr, 105(4), 380 - 3 An evaluation of the necessity of 24-hour incubation for oxacillin minimum inhibitory concentrations; Wasilauskas BL et al.; Current standards of the National Committee for Clinical Laboratory Standards (NCCLS) for microtube dilution recommend 24-hour incubation of staphylococci when testing for oxacillin/methicillin resistance . This study was conducted to quantify the need for this requirement . Standard 16-hour readings were compared with subsequent 24-hour readings of 515 fresh clinical isolates (256 Staphylococcus aureus, 259 coagulase-negative staphylococci) that were susceptible to oxacillin (microtube dilution minimum inhibitory concentration {MIC} < or equal to 2 micrograms/mL) after 16 hours . Five hundred two of the susceptible isolates (97.5%) were still susceptible at 24 hours . The remaining 13 isolates were resistant (MIC > 2 micrograms/mL) at 24 hours . Duplicate retesting, alternative method testing (Kirby-Bauer, E-Test) and mec A gene analysis were performed on these 13 isolates . All 13 isolates possessed the mec A gene . Four isolates always tested resistant (including all 16-hour repeat microtube dilution readings), and one isolate always tested susceptible . The remaining eight isolates produced variable results suggestive of an MIC very close to the resistance/susceptible break point . Overall , conversion from susceptible to resistant was entirely dependent upon the 16-hour MIC . There were 53 isolates with 16-hour MICS of 2 micrograms/mL . All 13 converters came from this group . No isolate with an oxacillin MIC < 2 micrograms/mL at 16 hours was resistant at 24 hours . Based on these results, the authors instituted a selective reincubation of only those staphylococcal isolates with oxacillin readings of 2 micrograms/mL at 16 hours. Eur J Biochem, 1996 Mar 15, 236(3), 904 - 10 Purification and characterisation of a plasmin-sensitive surface protein of Staphylococcus aureus; Hilden P et al.; Certain methicillin-resistant Staphylococcus aureus strains contain a 230-kDa cell-wall protein which is not present on the surface of other staphylococci . The presence of this 230-kDa protein is associated with a negative test result in commercial assays designed to detect fibrinogen-binding proteins and/or protein A on the staphylococcal surface . We have purified and partially characterised the 230-kDa protein from a lysostaphin digest of a non-agglutinating methicillin-resistant S . aureus strain . Partial amino acid sequence data obtained from the purified protein did not reveal any significant similarities to known proteins which indicates that the protein is novel . The 230-kDa protein was very sensitive to proteolysis; soluble plasmin, or plasmin formed on the bacterial-cell surface, rapidly degraded the 230-kDa protein to a 175-kDa form . The finding that the 230-kDa protein bound to lectins allowed its purification by affinity chromatography on immobilised wheat germ agglutinin . Furthermore, the degradation of the 230-kDa protein was associated with an increased adherence of non-agglutinating methicillin-resistant S . aureus cells to solid-phase fibronectin, fibrinogen or IgG. Clin Perform Qual Health Care, 1996 Apr-Jun, 4(2), 107 - 9 Staphylococcus cohnii: a case report on an unusual pathogen; Fernandes AP et al.; Coagulase-negative staphylococci have become increasingly important causes of infection in predisposed hosts . such as patients receiving immunosuppressive therapy and broad-spectrum antimicrobial drugs, patients who have prosthetic devices, or those who have prolonged hospital or intensive care unit stays . However, human infections caused by Staphylococcus cohnii rarely have been reported in the literature . In this report, we review the current literature and describe a 38 year-old immunosuppressed woman who developed catheter-related S . cohnii bacteremia . The case illustrates why microbiology laboratories under certain circumstances should identify coagulase-negative staphylococci to the species level . This information may be critical because it may allow clinicians to identify the source of the infecting organism and to choose appropriate antibiotics . Yet in this era of cost containment many laboratories may decrease costs by decreasing services, including species identification. J Antimicrob Chemother, 1996 Mar, 37(3), 445 - 56 Identification of mecA-related oxacillin resistance in staphylococci by the E test and the broth microdilution method; Petersson AC et al.; A set of 165 strains of different staphylococcal species, 67 Staphylococcus aureus, 71 novobiocin-sensitive coagulase-negative staphylococci (CNS) and 27 novobiocin-resistant CNS was used . The oxacillin and methicillin MICs were recorded after 24 and 42 h of incubation at 35 degrees C and at 30 degrees C . Significantly higher MICs were recorded at 30 degrees C compared with 35 degrees C . While a poor discrimination between mecA-positive and mecA-negative strains was obtained with methicillin, the oxacillin MICs enabled identification of resistant strains under certain conditions . The distribution of MICs differed between the three groups of species . Separation of uninduced mecA-positive (> or = 4.0 mg oxacillin/L) and mecA-negative (< or = 2.0 mg oxacillin/L) strains of S . aureus was only achieved with the E test and after 42 h of incubation . Oxacillin-induction yielded higher MICs for mecA-positive strains of S . aureus, and a separation from mecA-negative strains was achieved with the E test after 24 h and with the broth microdilution method after 42 h . Separation of mecA-positive and mecA-negative strains of novobiocin-sensitive CNS required agar supplemented with 5% blood, incubation of MIC trays and E test for 42 h, and species-specific oxacillin MIC breakpoints (S < or = 0.5 mg/L and R > or = 1.0 mg/L) . The mecA-positive and mecA-negative strains of novobiocin-resistant CNS were clearly separated after 24 h of incubation by either method. Pol Tyg Lek, 1996 Mar, 51(10-13), 145 - 7 {Pseudoaneurysm after anastomosis of a prosthesis with the femoral artery}; Witkowski M et al.; One of the complications of femoral artery synthetic prostheses are anastomotic aneurysms . In 1985-1993, 38 patients were treated for anastomotic aneurysms . Diagnosis was based on clinical and ultrasound examinations . A retrospective analysis has shown an occurrence of fever in 12 patients, and superficial pus oozing in 10 patients . Bacteriological cultures of intra-operatively collected specimens were positive in 25 patients . An infection with staphylococci was predominating . Repeated reconstructive surgery was performed within 6 months to 5 years after the primary one, mean 1.6 years . Aneurysm with the segment of prosthesis was excised and replaced by a fragment of new prosthesis (26 patients) or the vessel after excision of aneurysm was sutured with additional sutures on the whole anastomosis circumference (6 patients) . On 7 patients bypass grafting was performed with ligation of vessels supplying anastomotic aneurysm . In all patients flow drainage was used . Positive result of re-operation was achieved in 21 (68.4%) patients . Limb was amputated in 10 cases (26.3%), and 2 patients (5.3%) died . An analysis of the treated patients showed an important role of the anastomotic infection in the anastomotic aneurysms formation . The treatment should include prolonged antibiotic therapy prior to the operation . In case of reoperation possible infection should always be considered. Semin Respir Infect, 1996 Mar, 11(1), 54 - 61 Histology and microbiology of ventilator-associated pneumonias; Rouby JJ; A good knowledge of histological and bacteriological characteristics of experimental and human ventilator-associated bronchopneumonias (BPN) is of critical importance for the intensivist . BPN can be experimentally produced by intratracheal inoculation of microorganisms in high concentrations and ventilator-associated BPN by ventilating baboons with oleic-acid lung injury . Experimental ventilator-associated BPN is frequently polymicrobial, and bacterial lung burden increases with the severity of lung infection . In human ventilator-associated BPN, gross examination is of poor value for diagnosing lung infection . Four histologic categories of increasing severity have been described: bronchiolitis, focal bronchopneumonia, confluent bronchopneumonia, and lung abscess . Nonspecific inflammatory lesions are always associated with histologic lung infection: primary lung infection causes secondary inflammatory lung damage, whereas non-specific alveolar injury is rapidly superinfected when the lungs are mechanically ventilated . Infectious pulmonary lesions are disseminated within all pulmonary segments but preferentially found in the dependent segments . This fact suggests that ventilator-associated BPN has a bronchogenic origin and that gravity plays an important role in the dissemination of microorganisms within lung parenchyma . Ventilator-associated BPN is a nosocomial infection with a predominance of gram-negative bacteria, staphylococci species, and yeasts . It is frequently polymicrobial, and the lung bacterial burden depends on the histologic grade, the administration of topical and intravenous antibiotics, and the host's local antibacterial defenses . The bacterial complexity of human lung infection does not support the concept of a threshold for the diagnosis of nosocomial BPN . Intensivists should always keep in mind that human ventilator-associated BPN is a complex and rapidly changing entity. Arch Dis Child Fetal Neonatal Ed, 1996 Mar, 74(2), F99 - 104 Reservoirs of coagulase negative staphylococci in preterm infants; Eastick K et al.; This investigation was undertaken to determine the magnitude of, and interrelations between, reservoirs of coagulase negative staphylococci on infants' skin at various sites (including sites used for insertion of intravascular catheters) and in faeces during the first six months of life . Sites with large numbers of coagulase negative staphylococci were identified by sampling 16 skin sites and stools from 20 preterm neonates at 8-30 days of life . A more detailed survey of numbers and types of coagulase negative staphylococci in stool and at six skin sites of 10 preterm infants was then performed over the first six months of life . Isolates of coagulase negative staphylococci were collected and characterised by speciation, antibiotic susceptibility profiling, and plasmid restriction fragment length polymorphism analysis . Large, relatively stable reservoirs were identified in the faeces, around the ear, and in the axilla and nares . Skin on the forearm and leg, sites at which peripheral catheters are frequently sited, carried small unstable numbers of coagulase negative staphylococci, which were usually indistinguishable from coagulase negative staphylococci isolated from other body sites on the same baby . Contamination of catheter insertion sites with coagulase negative staphylococci from reservoir sites on the same baby could explain these observations . These data suggest that interventions reducing cross-contamination between sites on the same baby might be as important in preventing coagulase negative staphylococcal bacteraemias as measures taken to prevent cross infection between babies . Procedures which are likely to result in heavy coagulase negative staphylococcal contamination of the hands of healthcare staff, such as changing soiled nappies, should receive particular attention. Blood Coagul Fibrinolysis, 1996 Mar, 7 Suppl 1, S45 - 51 Bacteriology, safety and prevention of infection associated with continuous intravenous infusions; Mermel LA; There are over 50,000 intravascular catheter-associated bloodstream infections in the United States each year; globally, the number of these infections is likely to be much higher . At least half of these bloodstream infections are caused by staphylococci . The source of most pathogens causing endemic catheter-associated bloodstream infections is the catheter insertion site or the catheter hub, whereby microbes migrate into the bloodstream along the outside or inside of the catheter, respectively . The pathogenesis of epidemic intravascular catheter-related bloodstream infections is quite different . Epidemic bloodstream infections are due to manufacturer-related contamination or contamination that occurs at the location of catheter use, such as the hospital . These epidemics have most often been traced to contamination of intravenous solutions such as hyperalimentation or medications, blood products, contaminated cutaneous antiseptics or faulty decontamination of reusable devices . The prevention of infection