Anaesth Intensive Care, 1999 Oct, 27(5), 447 - 51 Pulsed-field gel electrophoresis is a useful tool in the monitoring of methicillin-resistant Staphylococcus aureus epidemic outbreaks in the intensive care unit; Cameron RJ et al.; We wished to determine how pulsed-field gel electrophoresis may be of use in monitoring methicillin-resistant Staphylococcus aureus (MRSA) outbreaks in the intensive care unit (ICU) . A retrospective epidemiological analysis was conducted . All 27 ICU patients and 11 patients from other hospital wards from whom MRSA was isolated over a one year period were included in the study . Seventeen of the 27 ICU MRSA isolates were analysed by pulsed-field gel electrophoresis for clonality and compared with the 11 other hospital isolates genotypes over the same period . During three MRSA outbreaks, five MRSA genotypes were identified in ICU whilst the same five genotypes and three additional were found in the rest of the hospital . Pulsed-field gel electrophoresis analysis was useful in identifying clonality of ICU MRSA infections and establishing that they were imported from hospital wards, rather than arising de novo in ICU . We were further able to identify clonal clusters within the unit linked by temporal and geographical proximity, suggestive of cross-infection . Pulsed-field gel electrophoresis typing might be additionally useful in tracing the source of human and/or environmental factors if a genotype were persistently identified. Scand J Immunol, 1999 Oct, 50(4), 363 - 70 Staphylococcus aureus Cowan strain 1 activation of B-chronic lymphocytic leukaemia cells augments the response to CD40 stimulation; Soderberg O et al.; The signals involved in regulating the proliferation, differentiation and survival of B-chronic lymphocytic leukemia (B-CLL) cells are fully understood . B-CLL cells have been found to respond poorly to various activation signals and only after successful Epstein-Barr virus (EBV) transformation has it been possible to maintain such cells in long-term cultures . In this work we describe a new method to activate and induce proliferation in B-CLL cells and to maintain such cells in long-term culture for longer than 1 month . We used a combination of protocols in an attempt to mimic some of the signals of a thymus-dependent immune response . The B-CLL cells were first activated with Staphylococcus aureus Cowan strain 1 (SAC) particles plus thioredoxin (Trx), followed by stimulation with interleukin (IL)-2 + Trx . This treatment primed the cells for further stimulation with anti-CD40 monoclonal antibody (MoAb) presented on irradiated CD32L cells (the CD40-system) or soluble CD40 Ligand, and a combination of Trx and cytokines (IL-4 + IL-10), which allowed the cells to be maintained for up to 1 month with preserved viability and a variable rate of proliferation . However, induced proliferation of the B-CLL cells was limited to approximately 1 month, suggesting that additional signals are required to facilitate further proliferation. FEMS Microbiol Lett, 1999 Oct 15, 179(2), 233 - 9 Potentiation of methicillin activity against methicillin-resistant Staphylococcus aureus by diterpenes; Nicolson K et al.; Totarol is a diterpene compound extracted from the totara tree . Totarol and eight other diterpenes were found to potentiate methicillin, one reducing the minimum inhibitory concentration of methicillin against resistant Staphylococcus aureus 256-fold . Totarol did not inhibit the synthesis of DNA or peptidoglycan in S . aureus, but reduced the respiration rate by 70% . Under potentiation conditions, diterpenes had only a slight effect on the respiration rate, but had a significant effect on expression of PBP 2a . We conclude that the primary staphylococcal target for totarol is the respiratory chain, but that potentiation of methicillin by diterpenes is by interference with PBP 2a expression. Med Pregl, 1999 Jun-Aug, 52(6-8), 267 - 9 {Effect of the "baby friendly" program on the number of neonatal infections at the maternity ward in Senta}; Molnar-Sabo I et al.; INTRODUCTION: The aim of this paper was to assess how the Baby Friendly Hospital Initiative affects occurrence, structure and outcome of neonatal infections at our department . MATERIAL AND METHODS: This retrospective study included all newborn infants born in 1995, when all the babies were at the neonatal ward and all the babies born in 1998, who were with their mothers . Newborn infants with low-birth-weight or shortened gestation were excluded . The assumption was that faster onset of lactation and thus breast-feeding decrease incidence of infections, but that there is an increased risk due to hygienic habits of mothers, especially those with no qualifications and difficult living conditions . The paper assesses the percentage of infections occurrence . RESULTS: Occurrence of infection was established clinically, whereas general signs of infection, as well as local signs of infection were confirmed by laboratory and bacteriological findings . Antibiotic therapy was applied . In great number of infections Staphylococcus aureus was isolated . In 1998 a certain increase of low-birth-weight and low gestation newborn infants was registered . In mature babies included into the Baby Friendly Program, number of infections has not changed, but the treatment was a little shorter . Infections were much more frequent in low-birth-weight and low gestation newborn infants . On the average the treatment in such cases was a little longer, but not only due to infection . DISCUSSION: Baby Friendly Hospital Initiative has not significantly affected the incidence of intrahospital infections in newborn infants . On the average the treatment in mature newborn infants was shorter, probably due to better lactation and transfer of immunoglobulins from mother to child . CONCLUSION: If Baby Friendly Hospital Initiative means adequate epidemiological supervision of mothers, this program does not significantly affect the risk from intrahospital infections. Microbiology, 1999 Sep, 145 ( Pt 9), 2497 - 505 Matrix-binding proteins of Staphylococcus aureus: functional analysis of mutant and hybrid molecules; Hartford O et al.; The fibrinogen-binding protein ClfA and the collagen-binding protein Cna are surface-associated adhesins of Staphylococcus aureus . ClfA has a dipeptide repeat region R composed mainly of serine and aspartate residues, more than 40 of which are required along with the 28-residue region W, the LPXTG motif and region M to display the ligand-binding region A on the cell surface in a functional form . Cna has a 61-residue region W and at least one 187-residue region B linking the collagen-binding region A to peptidoglycan . A cna mutant of S . aureus lacking region B was shown to bind collagen at the same level as wild-type Cna+ cells, indicating that region B is not necessary for ligand binding . Furthermore, altering the number of B repeats did not influence the level of collagen binding . In order to study the ability of C-terminal domains of Cna and ClfA to support functional ligand-binding activity of different adhesins, chimeric proteins were constructed and expressed in S . aureus . Surprisingly, the presence of a single Cna B domain and a nonapeptide linker located between ClfA region A and Cna region WM failed to support fibrinogen binding by S . aureus cells, despite the fact that ClfA region A was detected on the bacterial surface by immunoblotting . In contrast, the ClfA region A-Cna region B hybrid expressed as a recombinant protein in Escherichia coli did bind fibrinogen in Western ligand blots and in an ELISA-type assay . It is concluded that Cna region B cannot support functional display of ClfA region A on the bacterial cell surface . However, the ClfA dipeptide repeat region R and region WM did promote functional surface expression of the Cna collagen-binding domain in a hybrid Cna-ClfA protein. Int J Biol Macromol, 1999 Nov, 26(2-3), 167 - 71 Micropurification of beta- and gamma-crystallins from rabbit aqueous humor; Leone MG et al.; Soluble crystallins are normally present in the aqueous humor, originating from the lens, and their concentration may increase in certain conditions such as cataract, possibly contributing to aqueous outflow pathway obstruction, leading to glaucoma . Whether the stability and the tendency of aqueous crystallins to aggregate are different in patients with certain forms of open-angle glaucoma has not so far been established, mainly due to the lack of a suitable purification procedure from this fluid in which crystallins are present at very low concentration together with dozens of other proteins . About 4 microg each of beta- and gamma-crystallins were obtained from 20 ml of rabbit aqueous humor by C8 reversed-phase high-performance liquid chromatography (HPLC) and high-performance electrophoresis chromatography (HPEC) . The identity of the proteins was confirmed by amino acid analysis following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and electrophoretic blotting onto polyvinylidene fluoride membranes, with or without previous digestion with Staphylococcus aureus protease V8. Mol Gen Genet, 1999 Sep, 262(2), 323 - 31 Interactive regulatory pathways control virulence determinant production and stability in response to environmental conditions in Staphylococcus aureus; Lindsay JA et al.; The accessory gene regulator (agr) and staphylococcal accessory regulator (sar) loci are important regulators of toxin production in Staphylococcus aureus . In this study we examined how environmental conditions degree of aeration and salt concentration - affect the transcription and translation of mRNAs for alpha-haemolysin (Hla) and serine protease (Ssp) via these pathways and influence the stability of these proteins . Using Northern analysis, we have confirmed earlier observations that sarA is involved in the upregulation of RNAIII, the effector molecule encoded by the agr locus . However, this effect was abolished in highly aerated cultures . While sarA does appear to have an up-regulatory effect on hla transcription that is independent of agr, we propose that the PC1839 (sarA) mutant produces less alpha-haemolysin activity mainly as a result of post-translational inactivation by proteases . The most obvious phenotypic feature of PC1839 (sarA) is the upregulation of proteases . In this study we show that ssp is repressed by SarA at the transcriptional level . Western analysis using an anti-alpha-haemolysin antibody identified a major breakdown product that is only present in the supernatant of strains that are overexpressing serine protease . We have also confirmed that agr exerts a significant regulatory influence on hla at the level of translation, as well as transcription . Finally, the addition of salt upregulates ssp transcription and dramatically downregulates transcription of hla, and is an example of an environmental parameter that affects toxin production independently of agr and sarA . How environmental signals are transduced to control alpha-haemolysin and serine protease production, activity and stability at multiple levels are discussed. Mult Scler, 1999 Oct, 5(5), 327 - 34 Impaired interleukin-12 production in multiple sclerosis patients; Rohowsky-Kochan C et al.; Multiple Sclerosis (MS), a disease of the human central nervous system, is believed to be a T cell mediated autoimmune disorder with genetic and environmental influences . Interleukin-12 (IL-12), a proinflammatory cytokine produced primarily by antigen presenting cells is a potent inducer of interferon-gamma (IFN-gamma) and other Th1 cytokines that may play an important role in MS pathogenesis . We have investigated IL-12 production induced by the T cell independent pathway in untreated and IFN-beta treated MS patients, healthy individuals and other neurological disease (OND) patients in response to the human pathogen Staphylococcus aureus . We report that peripheral blood mononuclear cells (PBMC) from untreated MS patients produce normal amounts of the biologically active IL-12 p70 heterodimer but significantly less free IL-12 p40 heavy chain than PBMC from both healthy and disease controls when challenged in vitro with Staphylococcus aureus . Both mRNA expression of the inducible IL-12 p40 chain and protein levels were found to be reduced in untreated MS patients . No decrease in the production of the IL-12 p40 was seen in MS patients on IFN-beta therapy . The decreased production of IL-12 p40 heavy chain is not attributed to increased IL-10 secretion, a defect in the production of cytokines by macrophages or the number of cytokine producing cells . The factor(s) responsible for the decrease in p40 remain to be determined . Since IL-12 p40 antagonizes the biological activity of IL-12 in vitro and in vivo, identification of a defect in the 'natural' antagonist of IL-12, may provide the basis for immune therapy. J Infect Dis, 1999 Nov, 180(5), 1561 - 8 Influence of in vitro susceptibility phenotype against thrombin-induced platelet microbicidal protein on treatment and prophylaxis outcomes of experimental Staphylococcus aureus endocarditis; Dhawan VK et al.; Thrombin-induced platelet microbicidal protein-1 (tPMP-1) is a small, cationic staphylocidal peptide from rabbit platelets . In the current study, the outcomes of vancomycin treatment and prophylaxis were compared in experimental infective endocarditis (IE) caused by an isogenic Staphylococcus aureus strain pair differing in tPMP-1 susceptibility (tPMPS) or resistance (tPMPR) in vitro (ISP479C and ISP479R, respectively) . Vancomycin therapy (selected for its intrinsically slow bactericidal activity) reduced ISP479C (but not ISP479R) densities in vegetations compared with controls (P<.01) . In contrast, prophylactic administration of vancomycin yielded no differences in efficacies for the 2 challenge strains . These data suggest that the tPMPR phenotype in vitro has a negative effect on the antimicrobial therapy (but not the prophylaxis) of experimental S . aureus IE . These disparate results may be explained in part by the requirement for microbicidal effects in the treatment of established IE, whereas prophylactic efficacy depends more on growth inhibitory and antiadhesion effects. J Orthop Trauma, 1999 Sep-Oct, 13(7), 470 - 6 Influence of the design for fixation implants on local infection: experimental study of dynamic compression plates versus point contact fixators in rabbits; Arens S et al.; OBJECTIVES: Comparison of infection resistance after local bacterial challenge associated with two different designs for fixation implants: the conventional dynamic compression plate (DCP) and the point contact fixator (PC-Fix) . DESIGN: Randomized, prospective study in experimental animals . Grouped sequential experimental procedure . Observation time was twenty-eight days, with twenty animals per group . SETTING: Following surgery, animals were kept without restrictions in individual hutches . ANIMALS: Forty White New Zealand rabbits . Thirty-eight animals, nineteen per group, were included in the final evaluation . INTERVENTION: Under sterile conditions, specially manufactured titanium DCP or PC-Fix of identical dimensions were fixed to rabbit tibiae . After wound closure, different concentrations of Staphylococcus aureus, between 2 x 10(4) and 2 x 10(8) colony-forming units (CFU), were inoculated percutaneously at the implant site . MAIN OUTCOME MEASUREMENTS: Implants, underlying bone, and surrounding soft tissues were removed under sterile conditions and quantitatively evaluated for bacterial growth . Infection was defined as positive bacterial growth at the bone-implant interface . RESULTS: The overall infection rate was 45 percent . The infection dose of 50 percent (ID50) was 7.08 x 10(5) CFU for the DCP group and 8.51 x 10(6) CFU for the PC-Fix group . The infection rate was 63 percent (twelve of nineteen animals) for the DCP group and 26 percent (five of nineteen animals) for the PC-Fix group . This difference was statistically significant (p = 0.022) . CONCLUSIONS: After local bacterial challenge, we found a statistically significant difference in the infection rates depending on the implant design . The higher infection resistance associated with the PC-Fix design seems to be related to the reduced contact area at the bone-implant interface. J Antibiot (Tokyo), 1999 Jul, 52(7), 660 - 5 In vivo antibacterial activity of FK041, a new orally active cephalosporin; Tawara S et al.; The therapeutic activities of orally administered FK041 were evaluated in mouse models of systemic and local infections with a variety of bacteria and were compared with those of cefdinir (CFDN) and cefditoren pivoxil (CDTR-PI) . FK041 exhibited potent therapeutic activity against lethal systemic infections induced by intraperitoneally inoculated Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae with 50% effective doses (ED50) in the range of 0.20 to 0.36 mg/kg and was more active than CFDN and CDTR-PI . This result correlated well with its in vitro activity . The therapeutic effects of FK041 and reference drugs on murine local infections were evaluated in an in vivo pharmacokinetic model simulating human plasma concentrations for oral administration of 50 mg, 100 mg, and 200 mg . Against murine subcutaneous abscess induced by S . aureus, FK041 was as effective as CFDN and significantly more effective than CDTR-PI in reducing the number of recoverable viable bacteria in the skin at the infection sites . The efficacy of FK041 against murine pneumonia with H . influenzae was comparable to that of CDTR-PI and was superior to that of CFDN in reducing viable bacteria activity in the lungs . These results strongly suggest that FK041 has potential for clinical use against various bacterial infections. Arthritis Rheum, 1999 Sep, 42(9), 1823 - 7 Fcgamma receptor polymorphisms in Wegener's granulomatosis: risk factors for disease relapse; Dijstelbloem HM et al.; OBJECTIVE: Several studies have recently identified polymorphisms of receptors for the Fc fragment of IgG (FcgammaR) as genetic factors influencing susceptibility to multiple autoimmune and infectious diseases . This genetic predisposition could also influence the expression of Wegener's granulomatosis (WG), a systemic autoimmune disease with chronic nasal carriage of Staphylococcus aureus as an important risk factor for disease relapses . Therefore, we analyzed 3 functional FcgammaR polymorphisms from 91 patients with WG and 154 controls for a possible relationship with disease expression and occurrence of relapses . METHODS: FcgammaR phenotypes were determined using amplification of FcgammaR-genomic regions in allotype-specific polymerase chain reactions . Of particular interest in the analysis were 2 allotypic forms of FcgammaRIIa (R131 or H131) and 2 allotypic forms of FcgammaRIIIa (V158 or F158), all of which are functionally different . RESULTS: Analysis of FcgammaR phenotypes demonstrated that patients with WG were more prone to disease relapse in the first 5 years after diagnosis if they were homozygous for both the R131 form of FcgammaRIIa and the F158 form of FcgammaRIIIa (relative risk 3.3, 95% confidence interval 1.6-6.8) . These polymorphisms are both associated with decreased FcR-mediated clearance, which may be relevant to the chronic nasal carriage of S aureus . CONCLUSION: Both the R/H131 polymorphism of FcgammaRIIa and the V/F158 polymorphism of FcgammaRIIIa represent heritable risk factors for the development of disease relapses in WG. J AOAC Int, 1999 Sep-Oct, 82(5), 1171 - 4 An immunoassay method for rapid detection of Staphylococcus aureus in cosmetics, pharmaceutical products, and raw materials; Hughes D et al.; The TECRA Staphylococcus aureus Visual Immunoassay allows a presumptive positive or negative result for the presence of S . aureus to be obtained within 26 h, in contrast to 4-5 days by traditional cultural methods . Presumptive positive immunoassay results are confirmed by streaking the enrichment broth onto conventional agar media . A validation study was undertaken to compare the TECRA assay with a cultural reference method based on the Bacteriological Analytical Manual (8th Ed.), which is also consistent with U.S . Pharmacopoeia requirements . The products tested included a range of cosmetics (toothpaste, shampoos, conditioners, sunscreens, moisturizers, lip and eye creams) and pharmaceuticals (cough mixtures, laxatives, ulcer treatments, infant formulae, antiseptic cream), as well as some pharmaceutical ingredients . Samples were inoculated with S . aureus at 10-20 cfu/g, and then enriched for 24 h at 35 degrees-37 degrees C at a product-to-sample ratio of 1:100 . Two different enrichment broths were used for the study: Tryptone Soya Broth with 4% Tween 80 and Modified Letheen Broth . For both enrichment broths, results of the immunoassay and the reference method showed close correlation . The TECRA S . aureus Visual Immunoassay provides a rapid and convenient alternative to cultural methods and provides advantages to industry, such as greater speed of product and ingredient release and faster tracing of contamination problems . Because the immunoassay may be read either visually or with the aid of a plate reader, there is no need for an initial outlay on capital equipment . However, the assay can be automated if required. Vet Microbiol, 1999 Sep 15, 69(3), 217 - 24 Genotyping of Staphylococcus aureus isolated from bovine mastitis; Annemuller C et al.; The present study was designed to comparatively investigate 25 Staphylococcus aureus strains isolated from bovine subclinical mastitis . The S . aureus strains, obtained from six different farms at five locations in one region of Germany, were characterized by phenotypic and genotypic methods . The S . aureus could be identified and further characterized by their cultural, biochemical and hemolytic properties . To analyze the epidemiological relationship the isolates were subjected to DNA fingerprinting by macrorestriction analysis of their chromosomal DNA, by PCR amplification of the gene encoding the 16S-23S rRNA intergenic spacer, by PCR amplification of the gene encoding the IgG binding region and the X region of protein A and by amplifying, and subsequent, digestion of the gene encoding staphylococcal coagulase . The macrorestriction analysis revealed five DNA restriction patterns with DNA patterns I, III and IV occurring in three, four, and three different farms, respectively . In addition, clones with different DNA patterns could be found within one herd . The PCR products for the spacer DNA, the spa gene encoding the X region of protein A and the coa gene encoding coagulase corresponded mostly to the pattern observed by DNA fingerprinting . Amplification of the gene encoding the IgG binding region revealed sizes of 620 bp for 20 of the isolates and 280 bp for four isolates indicating, for the latter, a deletion of segments in this region . These findings show, that single, widely distributed clones seemed to be responsible for cases of bovine subclinical mastitis found in one region of Germany. Vet Microbiol, 1999 Sep 15, 69(3), 189 - 98 Comparison of Staphylococcus aureus recovered from personnel in a poultry hatchery and in broiler parent farms with those isolated from skeletal disease in broilers; Rodgers JD et al.; Personnel from one broiler hatchery, and workers on 18 separate broiler parent farms which supply the hatchery, were tested for hand and nasal carriage of Staphylococcus aureus . In both locations, nasal carriage of S . aureus was more common than hand carriage . A total of 63 S . aureus strains were characterised by biotyping, protein A analysis and pulsed field gel electrophoresis (PFGE) typing . Of these, 36 were recovered from broiler hatchery personnel, 14 from broiler parent farm personnel and 13 from cases of skeletal disease in commercial broilers . Biotyping and protein A analysis indicated that none of the strains recovered from hatchery personnel were of the poultry biotype, but that two strains recovered from the hands of two broiler parent farm personnel could be grouped together with 12/13 of strains recovered from skeletal disease in broilers, as poultry biotypes . PFGE-typing could not distinguish 9/13 strains recovered from skeletal disease in broilers and one of the strains from the broiler parent farm personnel from isolate 24 (I . 24), which is the predominant S . aureus strain type associated with clinical disease in N . Ireland broiler flocks . The present study found no evidence of nasal carriage of S . aureus strains of poultry biotype by humans . The finding of hand carriage by broiler parent farm personnel, suggests that handling by personnel may contribute to the dissemination of I . 24 or other S . aureus strains associated with skeletal disease in broilers. No To Shinkei, 1999 Sep, 51(9), 815 - 8 {Methicillin-resistant Staphylococcus aureus subdural abscess in an elderly patient with dementia}; Takahashi O et al.; A 74-year-old man was admitted to our hospital because of a fever of 38.2 degrees C and drowsiness . Two months before admission, he was admitted to another hospital with the diagnosis of Alzheimer's disease . One week before admission, he had a fever which was judged to be due to pyelonephritis . Because imipenem cilastatin and minocycline were not effective in relieving symptoms, he was transferred to our hospital . Methicillin-resistant Staphylococcus aureus (MRSA) was cultured from the blood, and vancomycin was started on the 5th hospital day . Because of the persistent fever and signs of inflammation, Gallium-scintigraphy was performed, showing abnormal accumulation in the left fronto-parietal region of the brain and the sacral region . Enhanced brain CT revealed a crescentic low density area and a fine, intense line of enhancement in the left fronto-parietal region . An emergency drainage of abscess was performed via single left fronto-parietal burr hole . A slightly yellowish, bloody, purulent fluid was obtained . The subdural space was irrigated with saline containing antibiotics and a drain was inserted . MRSA was cultured from the obtained fluid . The fever gradually subsided and drowsiness disappeared . He had had decubitus ulcer, stage I on the surface, in the sacral region, which later turned out to have unexpectedly deep undermining lesion reaching to periosteum . MRSA was cultured from this decubitus lesion . MRSA which entered into blood stream from the decubitus site might have been implanted in the subdural hematoma . Thus, subdural abscess should be kept in mind as an active differential diagnosis in elderly patients with fever and drowsiness. Diabet Med, 1999 Sep, 16(9), 767 - 71 Methicillin-resistant Staphylococcus aureus: an increasing problem in a diabetic foot clinic; Tentolouris N et al.; AIM: To study the prevalence of pathogenic organisms and the prevalence and outcome of methicillin-resistant Staphylococcus aureus (MRSA) infection in foot ulcers in diabetic patients . METHODS: A retrospective analysis of wound swabs taken from infected foot ulcers in diabetic patients, selected from an outpatient diabetic foot clinic . Seventy-five patients (79 ulcers) with positive wound swabs were included . Size of ulcer and time to healing, in particular for MRSA-infected ulcers, were measured in all patients . RESULTS: Gram-positive aerobic bacteria were the commonest micro-organism isolated (56.7%) followed by gram-negative aerobic bacteria and anaerobes (29.8% and 13.5%, respectively) . Of the gram-positive aerobes, S . aureus was found most frequently and 40% were MRSA . MRSA was isolated more commonly in patients treated with antibiotics prior to the swab compared to those who had not received antibiotics (P = 0.01) . Patients whose foot ulcers were infected by MRSA had longer healing time than patients whose ulcers were infected by methicillin-sensitive S . aureus (mean (range) 35.4 (19-64) and 17.8 (8-24) weeks, respectively, P = 0.03) . CONCLUSION: MRSA infection is common in diabetic foot ulcers and is associated with previous antibiotic treatment and prolonged time to healing . Further studies are required to assess the need for antibiotics in treating foot ulcers in diabetes and to assess the optimum therapeutic approach to this problem. J Immunol, 1999 Oct 15, 163(8), 4519 - 26 Nitric oxide participates in the recovery of normal jejunal epithelial ion transport following exposure to the superantigen, Staphylococcus aureus enterotoxin B; McKay DM et al.; Bacterial superantigens (SAgs) are potent T cell activators . Mice treated 4 h previously with the SAg, Staphylococcus aureus enterotoxin B (SEB), display reduced ion transport (assessed by short circuit current) responses to prosecretory stimuli, which normalize 24 h posttreatment . Here, mice were treated with SEB alone or in combination with an inhibitor of the inducible form of NO synthase (iNOS), l -NIL . Subsequently, jejunal iNOS expression was detected by immunohistochemistry, ion transport was evaluated in Ussing chambers, and serum levels of TNF-alpha and IFN-gamma were measured by ELISA . SEB-treated mice had increased epithelial iNOS immunoreactivity, and numerous iNOS-positive CD3+ T cells occurred in their mucosa and submucosa . Concomitant treatment with l -NIL did not affect the reduced short circuit current responsiveness to electrical nerve stimulation or the prosecretory agents, carbachol and forskolin, that occurred 4 h post-SEB (5 microgram) treatment . However, Isc responses in l -NIL- plus SEB-treated mice were still significantly reduced 24 h posttreatment, indicating a role for NO in the restoration of normal ion transport following exposure to SAgs . The prolongation of epithelial ion transport abnormalities correlated with elevated serum levels of TNF-alpha and IFN-gamma in mice treated 24 h previously with l -NIL plus SEB compared with those in controls and SEB-only-treated mice . Additionally, mice treated with l -NIL plus SEB and TNF-alpha- or IFN-gamma-neutralizing Abs displayed normal jejunal ion transport characteristics 24 h posttreatment . We conclude that NO mobilization is important in the homeostatic recovery response following immune stimulation by SAgs and that the beneficial effect of NO in this model system is probably via regulation of TNF-alpha and IFN-gamma production. Bioorg Med Chem Lett, 1999 Sep 20, 9(18), 2685 - 90 3D QSAR studies on new oxazolidinone antibacterial agents by comparative molecular field analysis; Pae AN et al.; Three-dimensional QSAR studies for two series of new oxazolidinone antibacterial agents were conducted using the comparative molecular field analysis (CoMFA) . In vitro activities (MICs) of the compounds against methicillin-resistant Staphylococcus aureus 88 (MRSA 88) exhibited a strong correlation with their steric, electrostatic factors and lipophilicities. Antimicrob Agents Chemother, 1999 Oct, 43(10), 2565 - 8 Treatment of experimental staphylococcal endocarditis due to a strain with reduced susceptibility in vitro to vancomycin: efficacy of ampicillin-sulbactam; Backo M et al.; We evaluated several 3-day antimicrobial regimens in the treatment of experimental endocarditis caused by an oxacillin-resistant Staphylococcus aureus strain exhibiting intermediate susceptibility in vitro to vancomycin (VISA) . Neither vancomycin alone nor trovafloxacin exhibited in vivo efficacy; addition of amikacin to vancomycin yielded a modest in vivo effect . In contrast, the combination of ampicillin and sulbactam was highly effective in vivo, causing a mean decrease in VISA vegetation densities of >5 log(10) CFU/g versus those of untreated controls. Antimicrob Agents Chemother, 1999 Oct, 43(10), 2404 - 8 Multiple novel inhibitors of the NorA multidrug transporter of Staphylococcus aureus; Markham PN et al.; The multidrug transporter NorA contributes to the resistance of Staphylococcus aureus to fluoroquinolone antibiotics by promoting their active extrusion from the cell . Previous studies with the alkaloid reserpine, the first identified inhibitor of NorA, indicate that the combination of a chemical NorA inhibitor with a fluoroquinolone could improve the efficacy of this class of antibiotics . Since reserpine is toxic to humans at the concentrations required to inhibit NorA, we sought to identify new inhibitors of NorA that may be used in a clinical setting . Screening of a chemical library yielded a number of structurally diverse inhibitors of NorA that were more potent than reserpine . The new inhibitors act in a synergistic manner with the most widely used fluoroquinolone, ciprofloxacin, by substantially increasing its activity against both NorA-overexpressing and wild-type S . aureus isolates . Furthermore, the inhibitors dramatically suppress the emergence of ciprofloxacin-resistant S . aureus upon in vitro selection with this drug . Some of these new inhibitors, or their derivatives, may prove useful for augmentation of the antibacterial activities of fluoroquinolones in the clinical setting. Perit Dial Int, 1999 Jul-Aug, 19(4), 376 - 9 Effect of duration of chronic peritoneal dialysis therapy on the development of peritonitis; Troidle L et al.; OBJECTIVE: Long-term chronic peritoneal dialysis (CPD) therapy has been associated with alterations in peritoneal membrane structure and peritoneal macrophage function . We thus reviewed our experience with the development of peritonitis among patients maintained on CPD therapy for various time periods to determine if the spectrum of organisms, rates of peritonitis, and outcome changed with the duration of CPD therapy . SETTING AND PATIENTS: Patients maintained on CPD therapy in our out-patient unit in New Haven, Connecticut . DESIGN: Retrospective review of the charts of patients maintained on CPD therapy (HomeChoice Cycler or Ultrabag, Baxter, McGaw Park, IL, U.S.A.) between 1 January 1997 and 31 March 1998 . These patients were divided into three groups: group 1, patients maintained on CPD therapy < or = 12 months; group 2, patients maintained on CPD therapy for 13-36 months; and group 3, patients maintained on CPD therapy for > or = 37 months . RESULTS: The study included 256 patients: 101 patients in group 1, 110 patients in group 2, and 45 patients in group 3 . All groups of patients were similar in age . There were significantly fewer Caucasians and fewer males in group 3 in comparison to groups 1 and 2 . The incidence of diabetes mellitus, coronary artery disease, and peripheral vascular disease was significantly lower among patients in group 3 in comparison to groups 1 and 2 . There were 155 episodes of peritonitis during the study period for an overall rate of 1 episode in 18.7 patient-months . The overall, gram-positive, and gram-negative rates of peritonitis were not significantly different among the patients in groups 1, 2, and 3 . There were more episodes of Staphylococcus aureus peritonitis among patients in group 3 in comparison to group 2 (1 episode in 59.6 vs 1 episode in 280.2 patient-months, respectively) . Two weeks after the development of peritonitis, 94.6% of the patients in group 3 continued CPD therapy, while 79.4% of the patients in group 1 continued CPD therapy (p < 0.05) . No patient in group 3 transferred to hemodialysis, while 10.3% and 8.2% of the patients in groups 1 and 2 transferred to hemodialysis (p < 0.05) . The death rate 2 weeks after the onset of peritonitis was 10.3%, 9.8%, and 5.4% in groups 1, 2, and 3, respectively (p = NS) . CONCLUSIONS: Despite the immunological and morphological changes that occur in the peritoneal cavity with increased time on CPD therapy, there was no difference in the overall, gram-positive, or gram-negative rates of peritonitis for patients maintained on CPD therapy for various time periods . Patients in group 3 continued CPD therapy more often than did patients in group 1 . Patients in group 3 transferred to hemodialysis less often than did the remaining patients in the study period . The incidence of death was not significantly different for the three groups of patients. Crit Care Med, 1999 Sep, 27(9), 1908 - 15 Effect of short-term enteral feeding with eicosapentaenoic and gamma-linolenic acids on alveolar macrophage eicosanoid synthesis and bactericidal function in rats; Palombo JD et al.; OBJECTIVES: Because vasoactive eicosanoids derived from arachidonic acid present in immune cell phospholipids promote lung inflammation in critically ill patients, novel experimental diets containing eicosapentaenoic acid from fish oil and gamma-linolenic acid from borage oil have been designed to limit arachidonic acid metabolism . However, excess dietary eicosapentaenoic acid impairs superoxide formation and bacterial killing by immune cells . The present study determined whether short-term enteral feeding with diets enriched with either eicosapentaenoic acid alone or in combination with gamma-linolenic acid would modulate alveolar macrophage eicosanoid synthesis without compromising bactericidal function . DESIGN: Prospective, randomized, controlled, blinded study . SETTING: University medical center . SUBJECTS: Adult male Sprague-Dawley rats . INTERVENTIONS: Rats underwent surgical placement of a gastroduodenal feeding catheter and were randomly assigned to receive one of three high-fat (55.2% of total calories), low-carbohydrate diets containing isocaloric amounts of lipids for 4 days . The control diet was enriched with linoleic acid, whereas the two test diets were low in linoleic acid and enriched with either 5 mole % eicosapentaenoic acid alone or in combination with 5 mole % gamma-linolenic acid . Alveolar macrophages were then procured to assess phospholipid fatty acid composition, eicosanoid synthesis after stimulation with endotoxin, superoxide formation and phagocytosis by flow cytometry, and killing of Staphylococcus aureus MEASUREMENTS AND MAIN RESULTS: Alveolar macrophage levels of arachidonic acid were significantly (p < .01) lower and levels of eicosapentaenoic and dihomo-gamma-linolenic acids were higher after feeding the eicosapentaenoic and gamma-linolenic acid diet vs . the linoleic acid diet . Ratios of thromboxane B2,/B3, leukotriene B4/B5, and prostaglandin E2/E1 were reduced in the macrophages from rats given either the eicosapentaenoic acid or eicosapentaenoic acid with gamma-linolenic acid diet compared with ratios from rats given the linoleic acid diet . Macrophages from rats given the eicosapentaenoic with gamma-linolenic acid diet released 35% or 24% more prostaglandin E1 than macrophages from rats given either the linoleic acid or the eicosapentaenoic acid diet, respectively . Macrophage superoxide generation, phagocytosis of opsonized zymosan, and killing of S . aureus were similar irrespective of dietary treatment . CONCLUSION: Short-term enteral feeding with an eicosapentaenoic acid-enriched or eicosapentaenoic with gamma-linolenic acid-enriched diet rapidly modulated the fatty acid composition of alveolar macrophage phospholipids, promoted a shift toward formation of less inflammatory eicosanoids by stimulated macrophages, but did not impair alveolar macrophage bactericidal function relative to responses observed after feeding a linoleic acid diet. Pathol Int, 1999 Jul, 49(7), 672 - 5 Marked histiocytosis in the portal tract in a patient with reactive hemophagocytic syndrome: An autopsy case; Terada T et al.; We report an autopsy case of reactive hemophagocytic syndrome with peculiar liver histology . A 71-year-old female was diagnosed as having acute myelogenous leukemia and treated with chemotherapy . During her course, methicillin-resistant Staphylococcus aureus (MRSA) was noted in blood culture and she was diagnosed as having MRSA sepsis . She died of respiratory failure 5 months after the onset of leukemia and 10 days after the MRSA sepsis . Ante-mortem liver function tests were within normal ranges . At autopsy, myeloblastic leukemia cells positive for CD13 were present in the bone marrow and, to a much lesser extent, in the spleen and liver . Numerous histiocytes of a bland appearance with erythrophagocytosis were noted in the bone marrow and spleen . The histiocytes were positive for CD68, but negative for S-100 and lysozymes . In the liver, many histiocytes of bland appearance with erythrophagocytosis and CD68 positivity were present in the portal tracts with no Kupffer cell hyperplasia . There were no hepatocellular degeneration, fatty changes or sinusoidal dilations . We consider that this histiocytosis was associated with MRSA infection and diagnosed this as infection-associated hemophagocytic syndrome . In previously reported cases, hemophagocytosis in hyperplastic Kupffer cells was the main liver change of reactive hemophagocytic syndrome . The present case suggests that marked histiocytosis in portal tracts only may be a main feature of liver changes in reactive hemophagocytic syndrome and that such cases may not show abnormal liver function tests. Nippon Jinzo Gakkai Shi, 1999 Aug, 41(5), 505 - 10 {A case of non-Hodgkin's lymphoma presenting with acute renal failure diagnosed by renal biopsy}; Yaomura T et al.; We report a case of non-Hodgkin's lymphoma (NHL) presenting with acute renal failure . A-56-year-old male was admitted to our hospital on October, 1997 with fever and renal dysfunction . Physical examination showed no abnormality except for hepatomegaly . Body surface lymphadenopathy was not observed . Computed tomography (CT) of the abdomen showed markedly enlarged kidneys bilaterally and a mass of soft tissue density, which was considered as a swelling lymph node, around the aortic artery . The renal biopsy revealed parenchymal involvement of the NHL cells without normal tubulo-interstitial structure, but the glomeruli were almost intact . Our case rapidly fell into oliguria and acute renal failure, hence needed hemodialysis . After chemotherapy was performed, his renal function gradually improved and the kidney became smaller on subsequent CT . Unfortunately, the patient happened to suffer from methicillin-resistant staphylococcus aureus (MRSA) infection in a neutropenic state and died . Necropsy revealed recovery of the renal interstitium without residual NHL cells . Renal lymphoma without any other organ or nodal involvement is a rare type of NHL, which considered primary renal lymphoma (PRL) . However, we believe this case to have been a result of lymphomatous infiltration of the kidneys in disseminated lymphoma. Infect Control Hosp Epidemiol, 1999 Sep, 20(9), 604 - 6 Prospective evaluation of a hospital epidemiologist's activities at a European tertiary-care medical center; Ruef C; OBJECTIVE: Assessment of the distribution of tasks and consultations provided by the hospital epidemiologist (HE) at University Hospital of Zurich (UHZ) . DESIGN: Prospective collection of data on hospital epidemiology consultations over a 3-year period (1995-1997) . Time spent per consultation and activities of infection control practitioners were not recorded . SETTING: A 1,040-bed tertiary-care university hospital in Zurich, Switzerland . RESULTS: Between January 1, 1995, and December 31, 1997, the HE received 1,660 requests for consultation . Advice or action was sought in the following areas: epidemiology (27.5% of requests); quality assurance, including antibiotic utilization and technology assessment (24.8%); infection control and practice guidelines (22.5%); disinfection and sterilization (11.6%); clinical infectious diseases (13.4%) . During 1997, 35% of epidemiology consults were related to methicillin-resistant Staphylococcus aureus and 5.8% to tuberculosis . Public or private hospitals not affiliated with UHZ requested 40% of all consults . CONCLUSIONS: This study shows that HEs are involved in many different activities . Only 27.5% of hospital epidemiology consultations were directly related to issues of epidemiology . Practical knowledge of the methodologies for continuous quality improvement and assessment of various new technologies is important for HEs . The results of this study may be useful in discussions between HEs and administrators about allocation of resources or issues of reimbursement. In Vitro Cell Dev Biol Anim, 1999 Sep, 35(8), 472 - 80 Staphylococcus aureus adherence to nasal epithelial cells in a physiological in vitro model; Hoefnagels-Schuermans A et al.; Nasal carriage of Staphylococcus aureus represents a risk factor for subsequent invasive infections and interpatient transmission of strains . No physiological in vitro model of nasal epithelial cells is available to study both patient- and bacteria-related characteristics and their interaction, leading to adherence and colonization . Starting with tissues from human nasal polyps, a confluent, squamous, nonkeratinized epithelium in collagen-coated 96-well microtiter plates was obtained after 14 d . This in vitro cell-layer was characterized histologically, ultrastructurally, and immunohistochemically and showed features that were indistinguishable from those observed in the squamous nonkeratinized epithelium found in the posterior part of the vestibulum nasi . Adherence experiments were performed with four different 3H-thymidine-labeled Staphylococcus aureus strains . The effect of bacterial inoculum size, temperature of incubation, and incubation medium were studied . The adherence results were found to be reproducible, reliable and sensitive, allowing detection of small quantitative differences in adherence between the Staphylococcus aureus strains . There was no significant difference in adherence at 23 degrees C and 37 degrees C, nor between the incubation medium M199 and phosphate-buffered saline . Plastic adherence could be reduced and standardized with use of siliconized tips and a constant bacterial inoculum volume of 100 microl/well . This physiological and reliable in vitro cell-culture model offers a unique opportunity to study Staphylococcus aureus adherence to squamous, nonkeratinized nasal epithelial cells and both patient and bacterial characteristics involved in this interaction. FEMS Microbiol Lett, 1999 Sep 15, 178(2), 271 - 5 Proteolytic cleavage of the repressor (BlaI) of beta-lactamase synthesis in Staphylococcus aureus; Lewis RA et al.; The proteolytic cleavage of BlaI was shown to correlate with beta-lactamase synthesis in Staphylococcus aureus . BlaI was found to be autoregulatory when expressed from the blaZ promoter . Insertion of a 10-bp linker into the SnaBI site of blaRI resulted in constitutive synthesis of beta-lactamase. J Bacteriol, 1999 Oct, 181(19), 6184 - 7 4,4'-diapophytoene desaturase: catalytic properties of an enzyme from the C(30) carotenoid pathway of Staphylococcus aureus; Raisig A et al.; Staphylococcus aureus synthesizes C(30) carotenoids . Their formation involves the introduction of three double bonds, which is catalyzed by a single enzyme . This enzyme, 4,4'-diapophytoene desaturase from S . aureus, was overexpressed in Escherichia coli and purified in one step by affinity chromatography, and then the protein was characterized with respect to substrate specificity, cofactor requirement, and oligomerization. J Bacteriol, 1999 Oct, 181(19), 5909 - 14 Expression of the Staphylococcus aureus UDP-N-acetylmuramoyl- L-alanyl-D-glutamate:L-lysine ligase in Escherichia coli and effects on peptidoglycan biosynthesis and cell growth; Mengin-Lecreulx D et al.; The monomer units in the Escherichia coli and Staphylococcus aureus cell wall peptidoglycans differ in the nature of the third amino acid in the L-alanyl-gamma-D-glutamyl-X-D-alanyl-D-alanine side chain, where X is meso-diaminopimelic acid or L-lysine, respectively . The murE gene from S . aureus encoding the UDP-N-acetylmuramoyl-L-alanyl-D-glutamate: L-lysine ligase was identified and cloned into plasmid vectors . Induction of its overexpression in E . coli rapidly results in abnormal morphological changes and subsequent cell lysis . A reduction of 28% in the peptidoglycan content was observed in induced cells, and analysis of the peptidoglycan composition and structure showed that ca . 50% of the meso-diaminopimelic acid residues were replaced by L-lysine . Lysine was detected in both monomer and dimer fragments, but the acceptor units from the latter contained exclusively meso-diaminopimelic acid, suggesting that no transpeptidation could occur between the epsilon-amino group of L-lysine and the alpha-carboxyl group of D-alanine . The overall cross-linking of the macromolecule was only slightly decreased . Detection and analysis of meso-diaminopimelic acid- and L-lysine-containing peptidoglycan precursors confirmed the presence of L-lysine in precursors containing amino acids added after the reaction catalyzed by the MurE ligase and provided additional information about the specificity of the enzymes involved in these latter processes. J Trauma, 1999 Sep, 47(3), 551 - 4 Anterior mediastinal abscess after closed sternal fracture; Cuschieri J et al.; BACKGROUND: Although sternal fractures after blunt chest trauma are markers for significant impact, the fracture itself is generally not associated with any specific wound complications . Mediastinal abscess and sternal osteomyelitis rarely occur after blunt trauma or cardiopulmonary resuscitation . Management of such complications is difficult, and requires a spectrum of operative procedures that range from simple closure to muscle flap reconstruction . METHODS: The trauma registry of a Level I trauma center was used to identify patients suffering a sternal fracture between January of 1994 and August of 1997 . Records were reviewed for the mechanism of injury, length of hospital stay, and posttraumatic mediastinal abscess . RESULTS: Twenty-six patients were identified with sternal fracture . No clinically significant cardiac or aortic complications were noted . Three patients, all with a history of intravenous drug abuse and requiring central venous access in the emergency room, developed methicillin resistant Staphylococcus aureus mediastinitis . Sternal re-wiring and placement of an irrigation system successfully treated all three patients . CONCLUSION: Posttraumatic mediastinal abscess is an uncommon complication of blunt trauma in general and sternal fracture in particular . It can be recognized by the development of sternal instability . Risk factors include the presence of hematoma, intravenous drug abuse, and source of staphylococcal infection . Treatment with early debridement and irrigation can avoid the need for muscle flap closure. South Med J, 1999 Sep, 92(9), 909 - 11 Smitten by a kitten; Israeli E et al.; Mammalian bite wounds are commonly encountered in the emergency department . When patients come early (<8 hours after injury), local infection is not usually evident . At this stage, the issue of providing prophylactic antibiotic therapy arises . We report a complication of a cat bite to the hand in a previously healthy 32-year-old man . This patient did not seek medical treatment immediately after the cat bite, and distinct local infection did not develop . Nevertheless, his course was complicated with acute Staphylococcus aureus endocarditis . We discuss the common pathogens involved in a cat bite infection, including S aureus, and delineate the indications for prophylactic antibiotic therapy after a mammalian bite wound. Infect Immun, 1999 Oct, 67(10), 5541 - 4 Alpha-toxin damages the air-blood barrier of the lung in a rat model of Staphylococcus aureus-induced pneumonia; McElroy MC et al.; We have shown that injury to alveolar epithelial type I cells may account, in part, for damage to the air-blood barrier of the lung in a rat model of Staphylococcus aureus pneumonia . We have also shown that alpha-toxin is an important cause of damage to the air-blood barrier; however, our data suggest that the toxin is not acting directly on alveolar type I cells. Infect Immun, 1999 Oct, 67(10), 5001 - 6 Development and characterization of a Staphylococcus aureus nasal colonization model in mice; Kiser KB et al.; Staphylococcus aureus nasal carriage is a risk factor for infection in humans, particularly in the hospital environment . Attenuation of carriage has proven effective in reducing the prevalence of infection in some high-risk groups . To study staphylococcal factors that influence nasal colonization, a mouse model of S . aureus nasal colonization was developed . Mice were inoculated intranasally with S . aureus Reynolds, and nasal carriage was evaluated by quantitating cultures of the nasal tissues from mice sacrificed at various time points after inoculation . The majority of mice inoculated with 10(8) CFU of S . aureus maintained nasal carriage for at least 20 days . Nasal colonization rates were similar for inbred (BALB/c and C57BL/6) and outbred (ICR) mice . Colonization was not affected by mouse passage of strain Reynolds . Lower inoculum doses (<10(7) CFU) resulted in reduced colonization after 7 days . However, mice given streptomycin in their drinking water developed long-term carriage of S . aureus, and they were colonized with inocula as low as 10(5) CFU . Nasal colonization was also established with two other S . aureus strains (one strain each of human and murine origins) . S . aureus recovered from the nares of experimentally colonized mice expressed high levels of capsule, and the ability of a capsule-defective mutant to persist in the nares was reduced in comparison to that of the parent strain . This nasal colonization model should prove useful for studies of factors that mediate S . aureus colonization and for assessment of targets for antimicrobial intervention or vaccine development. Jpn J Thorac Cardiovasc Surg, 1999 Aug, 47(8), 368 - 76 Computerized antibiogram for methicillin-resistant Staphylococcus aureus in chest surgery; Yoshida J et al.; BACKGROUND: An increasing number of cases of postoperative morbidity involving methicillin-resistant Staphylococcus aureus have been reported in thoracic surgery . To prevent its outbreak, cluster analysis using a personal computer was employed . METHODS: A total of 120 patients undergoing operations on the lung and mediastinum were included into this study . Materials were isolates of methicillin-resistant Staphylococcus aureus newly recovered from across the hospital . The cluster analysis used antimicrobial susceptibility in 12 drugs, which were categorically valued to produce Euclidean distance to form clusters of similarity . RESULTS: Six of the 120 patients were found to be positive for the microbe before or after thoracotomy . A total of two patients (1.7%) became symptomatic postoperatively, i.e., one of four preoperatively-positive patients and one of two postoperatively-positive cases . The analysis suggested that preoperative patients shared the strains in the same non-surgical ward . DISCUSSION: A computerized antibiogram does not always strictly type Staphylococcal strains but has advantages in typing with ease and at decreased cost . The current analysis suggested that patient harboring the strains migrated across wards . CONCLUSION: Computerized antibiograms for Staphylococcal strains may assist to prevent an outbreak of their infection in chest surgery. Rev Soc Bras Med Trop, 1999 Jul-Aug, 32(4), 395 - 400 {A longitudinal study of healthy Staphylococcus aureus carriers among the students of a nursing aide course}; Santos BM; The objective of the present study was to determine the extent of colonization by Staphylococcus aureus and the evolution of carrier status among students of a technical nursing course during their professional training . Forty students participated in the study, samples were collected from their nasal cavity and right and left hands at six different times during the technical nursing course . Nineteen students (45.2%) were found to be occasional carriers, 12 (28.6%) were intermittent carriers, 6 (14.3%) were persistent carriers, and 5 (11.9%) were non-carriers, showing that colonization did not increase during the course . Twenty-four of them (57.1%) did not perform activities related to nursing before or during the course, whereas 18 (42.9%) performed them. Acta Derm Venereol, 1999 Sep, 79(5), 360 - 2 Bactericidal activity of manganese and iodide ions against Staphylococcus aureus: a possible treatment for acute atopic dermatitis; Inoue T et al.; We reported previously that balneotherapy using Kusatsu hot-spring water is useful for controlling the skin symptoms of acute flares/exacerbations of refractory cases of atopic dermatitis . As Staphylococcus aureus on the skin surface decreased in number or disappeared after balneotherapy, the hot-spring water was suspected to act against the microorganism . The hot-spring water showed strong bactericidal activity against S . aureus in vitro . In order to clarify the mechanism further, the bactericidal activity of the hot-spring water was examined by adding back cations and anions in same concentrations as those in the original hot-spring water, one at a time to cation- and anion-exchanged hot-spring water . The findings clearly demonstrated that the bactericidal activity was expressed by manganese and iodide ions in acidic conditions (pH 2.0-3.0) . Thus, the probable mechanism for the improvement of skin manifestations through Kusatsu balneotherapy is the bactericidal activity of the hot-spring water against S . aureus . When added to water acidified with sulphuric acid (pH 2.0-3.0) a synergistic effect of the 2 ions was observed, so that an anti-staphylococcal effect was obtained even at low concentrations (1 mg/kg) . Acidic solutions containing manganese and iodide ions may thus be clinically useful for treating skin conditions caused by S . aureus. J Neurosurg Sci, 1999 Mar, 43(1), 63 - 7 Cervical spinal epidural abscesses . A report on five cases; Piccolo R et al.; The authors report a series of five cases of non tuberculous cervical spinal epidural abscesses . There were neither patients suffering from immunodeficiency syndromes nor drug addicts; all the patients were in their seventh decade; two patients were affected by diabetes mellitus refractory to medical treatment . Retropharyngeal abscess was the main etiological risk factor (two cases); Staphylococcus aureus was cultured in two cases . Gadolinium MRI was necessary for a preoperative diagnosis, planning surgical approach and postoperative prognosis . Surgical debridement was performed via an anterior approach in those cases where the collection was located lower than C4 and did not span more than three vertebral segments; posterior approach, via a laminectomy, was performed in a case of C1-C2 location of the lesion and in a case of involvement of the whole cervical spine . Surgical results were poor in those patients affected by diabetes mellitus, a lesion involving the high cervical segments (higher than C4) or a lesion spanning more than three levels . Medical treatment with MRI follow-up was not undertaken in any of the patients and we opted for surgical drainage in all the cases due to the possibility of a sudden neurological deterioration, caused both by spinal cord mechanical compression and vascular compromission. Biochemistry, 1999 Sep 21, 38(38), 12296 - 304 Cooperative structural dynamics and a novel fidelity mechanism in histidyl-tRNA synthetases; Qiu X et al.; The crystal structure of the Staphylococcus aureus histidyl-tRNA synthetase apoprotein has been determined at 2.7 A resolution . Several important loops in the active site either become disordered or adopt very different conformations compared to their ligand-bound states . These include the histidine A motif (Arg257-Tyr262) that is essential for substrate recognition, a loop (Gly52-Lys62) that seems to control the communication between the histidine and ATP binding sites, the motif 2 loop (Glu114-Arg120) that binds ATP, and the insertion domain that is likely to bind tRNA . These ligand-induced structural changes are supported by fluorescence experiments, which also suggest highly cooperative dynamics . A dynamic and cooperative active site is most likely necessary for the proper functioning of the histidyl-tRNA synthetase, and suggests a novel mechanism for improving charging fidelity. Vestn Khir Im I I Grek, 1999, 158(1), 45 - 8; discussion 48-9 {The information value of the biological properties of the causative agent in the prognosis of the duration of the course of suppurative inflammatory diseases of staphylococcal etiology}; Deriabin DG et al.; The informative value of biological characteristics of Staphylococcus aureus reflecting the ability of microbes to inactivate certain factors of natural resistance (lysozyme, complement, immunoglobulins, bactericidal component of interferon) was established in determining the duration of the caused by them pyo-inflammatory diseases such as postinfection abscesses, lactation mastitis . On this basis the heterogeneous successive procedure of pattern recognition was used to develop algorithms allowing prognosis of the duration of pyo-inflammatory diseases of staphylococcal etiology in 84.6-94.4% of the patients after the quantitative and qualitative assessment of the biological properties of the pathogen with not less than 90% reliability. Biochem Biophys Res Commun, 1999 Sep 24, 263(2), 301 - 7 Mechanism of antibacterial and degradation behavior of a chlorinated isoxazolylnaphthoquinone; Bogdanov PM et al.; The chemical stability of 3-chloro-2-hydroxy-(3, 4-dimethyl-5-isoxazolyl)-1,4-naphthoquinon-4-imine (ClQ(1)), a new potential antimicrobial agent was analyzed at different pH values by first-derivative spectroscopy . The degradation of ClQ(1) followed a pseudo-first-order kinetics in aqueous media at different pH values . The interaction of antibiotics with respiratory chain of Staphylococcus aureus generates superoxide anion, an oxygen radical capable of producing damage to the bacteria . The performed assays have demonstrated that ClQ(1) presents higher activity and toxic oxidant generation at pH 5.0 than at pH 7.5 . In addition, the antibacterial activity of other halogenated isoxazolylnaphthoquinones was also studied in different collection and clinical strains which presented the following decreasing activity, ClQ(1) > BrQ(1) > DClQ(1) whereas DBrQ(1) did not show inhibition properties . The antibacterial and stability properties evidenced by ClQ(1) are so important that must be taken into account when new alternative treatments against beta-lactamase-positive S . aureus strains are investigated . Acta Crystallogr D Biol Crystallogr, 1999 Sep, 55 ( Pt 9), 1626 - 9 Crystal engineering: deletion mutagenesis of the 24 kDa fragment of the DNA gyrase B subunit from Staphylococcus aureus; Dale GE et al.; The 24 kDa fragment of DNA gyrase B from Staphylococcus aureus was expressed in Escherichia coli and purified for crystallization . Crystals of the wild-type protein grew in the presence of cyclothialidine but proved difficult to reproduce . In order to improve the crystallization, the flexible regions of the protein were deleted by mutagenesis . The mutant proteins were analyzed by differential scanning calorimetry and the most stable mutants produced crystals . It was possible to reproducibly grow single well defined crystals in the microbatch system which belonged to the space group C2 and diffracted isotropically to approximately 2 A resolution. Eur Respir J, 1999 Jul, 14(1), 113 - 7 Exhaled nitric oxide in patients with Wegener's granulomatosis; Haubitz M et al.; In Wegener's granulomatosis (WG), a pathogenic role of infections, in particular of a chronic colonization of the nasal mucosa with Staphylococcus aureus, has been postulated . Nitric oxide (NO), which is thought to play a role in primary host defence and inflammation, is produced endogenously within the respiratory tract, mainly from the paranasal sinuses . In order to further characterize its role in WG, nasal and pulmonary NO excretion in WG patients in comparison to healthy volunteers was measured . Seventeen patients with WG were included in the study . Five patients had active disease (bloody rhinitis with ulceration and crusting) and immunosuppressive therapy (IST), and 12 were in remission (six with, and six without, IST) . S . aureus was found in the swabs of all patients with active WG and in three patients in remission . NO was measured in exhaled gas using a chemiluminescence analyser . The NO excretion rate in nasally sampled gas was significantly reduced (p<0.05) in patients with active WG ((mean+/-SD)102+/-100 nL x min(-1)) compared to healthy controls (299+/-13 nL x min(-1)), and patients in remission (281+/-86 nL x min(-1) with IST, 280+/-133 nL x min(-1) without IST) . Pulmonary NO excretion in active or nonactive WG patients did not significantly differ from that of healthy volunteers (48+/-21 nL x min(-1)) . These results demonstrate a reduced nasal NO excretion in active Wegener's granulomatosis . This may be caused by destruction and/or functional impairment of sinus epithelium . The reduced NO concentration may well compromise host defence in the upper airways, thus contributing to colonization with Staphylococcus aureus and further promoting Wegener's granulomatosis. Plasmid, 1999 Sep, 42(2), 144 - 9 Rapid method for the identification of essential genes in Staphylococcus aureus; Xia M et al.; A strategy based on a vector host-dependent for autonomous replication, pSA3182, was utilized both for the rapid screening for Staphylococcus aureus genes essential for cell viability and for the introduction of specific polarity-neutral deletions in nonessential genes . The results obtained support the use of pSA3182 for both purposes . Plasmid, 1999 Sep, 42(2), 134 - 8 Comparative analysis of staphylococcal plasmids carrying three streptogramin-resistance genes: vat-vgb-vga; Allignet J et al.; Several staphylococcal plasmids (26-45 kb) carry all three streptogramin-resistance (Sg(R)) genes, vat, vgb, and vga . Seven such plasmids harbored by independent strains belonging to three taxa (Staphylococcus aureus, S . simulans, and S . cohnii subsp . urealyticum) were compared and the deleted derivative of one of them, pIP680 (11.3 kb), carrying the three streptogramin-resistance genes was sequenced . The seven native plasmids had in common a 12.1-kb part cocarrying the three Sg(R) genes . Sequence analysis of pIP680 revealed that the simultaneous presence of these three genes has probably resulted from cointegration of two plasmids: (i) a pAMbeta1-like plasmid harboring vat-vgb and whose replication gene has been inactivated by an IS257 insertion and (ii) a functional vga plasmid whose replication is similar to that of two staphylococcal plasmids, pSX267 and pSK41 . Mayo Clin Proc, 1999 Sep, 74(9), 885 - 9 Pseudo-outbreak of methicillin-resistant Staphylococcus aureus; Ender PT et al.; OBJECTIVE: To determine whether a high rate of methicillin-resistant Staphylococcus aureus at our institution was due to laboratory misclassification and to evaluate the effect of this misclassification . MATERIAL AND METHODS: We evaluated all S . aureus isolates identified at our institution during a 60-day period in 1997 . Automated susceptibility test results (using the Vitek system) from our clinical microbiology laboratory and an independent laboratory were compared with oxacillin agar screen plate results at both laboratories . Isolates with discordant results for susceptibility to oxacillin were tested by broth microdilution minimal inhibitory concentrations and for the presence of the mecA gene . RESULTS: Eighteen (72%) of the 25 organisms (obtained from 17 patients) found to be resistant to oxacillin by the Vitek system at our institution were susceptible by the oxacillin agar screen . Discordant isolates tested by broth microdilution minimal inhibitory concentrations and for the mecA gene were found to be oxacillin susceptible and mecA gene negative . Thus, at our hospital, almost three fourths of the organisms initially identified as methicillin-resistant S . aureus by the Vitek system were actually susceptible to oxacillin . This misclassification resulted in needless infection control measures and unnecessary vancomycin use . CONCLUSION: Hospitals that use only automated susceptibility testing for S . aureus should periodically validate their results with additional testing. Z Naturforsch {C}, 1999 Jul-Aug, 54(7-8), 549 - 53 Synergism between ethanolic extract of propolis (EEP) and anti-tuberculosis drugs on growth of mycobacteria; Scheller S et al.; Ethanolic extract of propolis exerts a strong anti-bacterial activity, in addition to antifungal, antiviral and antiprotozoal properties . In previous studies from these laboratories we have demonstrated that the intensity of the bactericidal activity of EEP is correlated with the virulence of the mycobacteria tested, and that EEP has a synergistic effect with antibiotics on growth of staphylococcus aureus . In the present study we investigated whether the same synergism and correlation exists between EEP and some anti-tuberculosis drugs on tuberculosis mycobacteria with different degrees of virulence . Six standard strains and 11 wild strains of mycobacteria were exposed for 30 days to EEP, with or without streptomycin, rifamycin, isoniazid or ethambutol . Out of the 17 strains, 8 were resistant to at least two standard antibiotics, and were considered "multi-resistant strains" . The rest were either susceptible or resistant to only one of the antimycobacterial drugs . Antagonism was recorded only in one case, when Staphylococcus aureus were treated with a mixture of EEP and ethambutol, suggesting that a chemical bond could have been formed between this anti-tuberculosis antibiotic and one of the active components of the ethanol extract of propolis. Zhonghua Gan Zang Bing Za Zhi, 1999 Jun, 7(2), 77 - 9 {Apoptosis of peripheral blood lymphocytes of patients with chronic hepatitis B and clinical significance}; Ji W et al.; OBJECTIVE: To explore the effects of apoptosis of peripheral blood lymphocytes (PBLs) of patients with chronic hepatitis B (CHB) on the persistent infection of hepatitis B virus (HBV) . METHODS: The PBLs of 15 patients with CHB were isolated and cultured with or without Staphylococcus aureus enterotoxin B (SEB; 0.2 mg/L), or in the presence or absence of recombinant HBcAg (rHBcAg; 1.0 mg/L) for 48 hours in vitro . After incubations, the cells were harvested by centrifugation and then apoptosis of the PBLs was studied by staining with fluorescent dyes YOPRO-1 and Hoechst33342 . RESULTS: The percentage of apoptotic cells of PBLs of patients with CHB was significantly higher than in normal controls (P < 0.01) in the presence or absence of SEB or rHBcAg, but the ratio of apoptotic cells in rHBcAg-stimulated PBLs of the patients was the highest, reaching (24.6 +/- 6.1)% . The patients with seropositive for HBeAg had higher percentage of apoptotic cells in their cultured PBLs than those with seronegative for HBeAg had, but the ratio of apoptotic cells in cultured PBLs of the patients with chronic heavy and severe hepatitis B was lower than that of the patients with chronic mild and moderate hepatitis B (P < 0.01) . CONCLUSION: Activation-induced cell death of PBLs of the patients with CHB may be related to persistent infection of HBV, and assay of apoptosis of PBLs of patients with CHB may provide valuable information about pathogenesis of CHB. J Clin Microbiol, 1999 Oct, 37(10), 3411 - 4 Use of multiplex PCR to detect classical and newly described pyrogenic toxin genes in staphylococcal isolates; Monday SR et al.; Staphylococcus aureus may contain one or more genes that encode a variety of immunomodulatory pyrogenic toxins (PTs), including the staphylococcal enterotoxins and toxic shock syndrome toxin (TSST) . The PTs interact with several cellular targets to produce disease, such as food poisoning and toxic shock syndrome . At present, nine serologically distinct enterotoxins and one immunoreactive form of TSST have been identified and characterized . As isolates of S . aureus are further assessed, it is anticipated that this number will increase . To facilitate screening, a multiplex PCR was designed to simultaneously determine which of these 10 currently known PT genes an individual S . aureus isolate possesses . We show here, using S . aureus isolates with characterized PT phenotypes, that this novel PCR technique reliably detects each of the known PTs in a single reaction. J Clin Microbiol, 1999 Oct, 37(10), 3198 - 203 Genotyping of epidemic methicillin-resistant Staphylococcus aureus phage type 15 isolates by fluorescent amplified-fragment length polymorphism analysis; Grady R et al.; Fluorescent amplified-fragment length polymorphism (FAFLP) analysis was investigated for its ability to identify and subtype isolates of an epidemic methicillin-resistant phage type of Staphylococcus aureus, EMRSA-15 . These isolates were also characterized by PCR-restriction fragment length polymorphism (PCR-RFLP) of the coagulase gene and pulsed-field gel electrophoresis (PFGE) . For FAFLP, DNA was double digested with restriction enzymes ApaI plus TaqI or EcoRI plus MseI . Site-specific adaptors were ligated to one or the other set of restriction fragments, and PCR amplification was carried out with adaptor-specific primers . Amplified fragments separated on an ABI 377 automated sequencer and analyzed with Genescan version 2.1 software generated FAFLP profiles for all the isolates . The presence or absence of fragments was scored, similarity coefficients were calculated, and UPGMA (unweighted pair group method using arithmatic averages) cluster analysis was performed . Either enzyme-primer combination readily differentiated EMRSA-15 from other methicillin-resistant S . aureus (MRSA) isolates and also revealed heterogeneity within the phage type . The discriminatory power of FAFLP was high . By combining both enzyme-primer data sets, 24 isolates were divided into 11 profiles . PCR-RFLP did not discriminate among these EMRSA-15 isolates . PFGE could discriminate well between isolates but was not as reproducible as FAFLP . All S . aureus and MRSA isolates in this study were typeable by FAFLP, which was easy to perform, robust, and reproducible, with evident potential to subtype MRSA for purposes of hospital infection control. J Clin Microbiol, 1999 Oct, 37(10), 3133 - 40 Follow-up of Staphylococcus aureus nasal carriage after 8 years: redefining the persistent carrier state; VandenBergh MF et al.; Studies of Staphylococcus aureus nasal carriage have distinguished three carriage patterns: persistent, intermittent, and noncarriage . The criteria used to identify these carriage patterns have been inconsistent . In 1988 the S . aureus nasal carrier index, i.e., the proportion of nasal swab specimen cultures yielding S . aureus, was determined for 91 staff members of various departments of a large university hospital by obtaining weekly nasal swab specimens for culture over a 12-week period . Thirty-three (36%) persons had carrier indices of 0.80 or higher, 15 (17%) had indices between 0.1 and 0.7, and 43 (47%) had indices of zero . In 1995, 17 individuals with carrier indices of 0.80 or higher in 1988 were available for reexamination . For 12 (71%) of these individuals, S . aureus was again isolated from a single nasal swab, i.e., from each individual with a 1988 carrier index of 1.0 but from only half of those with indices below 1.0 . Genotyping (by randomly amplified polymorphic DNA analysis and pulsed-field gel electrophoresis) of all S . aureus strains showed that strains isolated from only three individuals, all with 1988 carrier indices of 1.0, in 1988 and 1995 showed genetic similarity . In conclusion, persistent S . aureus nasal carriage is a unique characteristic of a fraction of the population, and the attribute "persistent" should be confined to those individuals for whom serial nasal swab specimen cultures consistently yield S . aureus. Int J Dermatol, 1999 Aug, 38(8), 582 - 6 Characteristics of bacterial skin infections in children compared to adults at a tertiary dermatologic center; Sugeng MW et al.; BACKGROUND: Bacterial skin infections in children and adults are caused by different organisms with different antimicrobial susceptibility . METHODS: A comparative retrospective study was carried out on 233 adults and 53 children with bacterial skin infections . Skin swab cultures and sensitivity tests were performed using standard methods . Statistical analysis was performed using the Pearson chi-squared and Fisher tests . A P value of < 0.05 was considered to be significant . RESULTS: Primary and secondary skin infections occurred in equal proportions in children, whereas secondary skin infections were more common in adults (70.8%) . Staphylococcus aureus was the main cause of skin infections, particularly in children (72.6%) . S . aureus in children and adults was highly susceptible to cloxacillin, cephalexin, chloramphenicol, neomycin, cotrimoxazole, and clindamycin, moderately susceptible to erythromycin, and insensitive to tetracycline, ampicillin, and penicillin . CONCLUSIONS: It is important to monitor the trends of bacterial infections and their antibiotic susceptibility as this can assist medical practitioners in their choice of antimicrobial therapy . Such monitoring will also help to detect the emergence of resistant bacterial strains and caution us to take care in the use of certain drugs. J Biotechnol, 1999 Aug 20, 73(2-3), 181 - 4 Expression of a clone specific DNA sequence from Staphylococcus aureus in Escherichia coli; el-Adhami W; Staphylococcus aureus produces a large number of factors thought to contribute to virulence, although the precise role of some of these individual factors is not clearly defined . To investigate whether specific virulence factors might be responsible for the selection and dominance of certain genotypes of methicillin- and multiply resistant S . aureus (MRSA), the method of subtractive hybridisation was used to identify conserved DNA sequences associated with the clinical, clonal populations of S . aureus . The findings described in this report indicate that the method of subtractive hybridisation is a valuable tool to identify clone specific virulence factors, which might be of potential as diagnostic markers and as alternative vaccine targets. Undersea Hyperb Med, 1999 Fall, 26(3), 169 - 74 Therapy with hyperbaric oxygen and cefazolin for experimental osteomyelitis due to Staphylococcus aureus in rats; Mendel V et al.; Hyperbaric oxygen (HBO2) is used as adjunctive therapy for chronic osteomyelitis, yet its efficacy remains controversial . A recently developed rat model for osteomyelitis due to Staphylococcus aureus was used to compare the results of treatment with HBO2, cefazolin, a combination of both, or no treatment . For the induction of tibial osteomyelitis, S . aureus was inoculated into the medullary cavity . Arachidonic acid was used as the sclerosing agent . With that procedure, an infection rate of 96% was attained . For long-term antibiotic treatment, a port system was developed and implanted . Hyperbaric treatment alone reduced the colony-forming units (CFU) from 2.9 x 10(6) to 6.2 x 10(5) x g(-1) of tibial bone . The effect on the infection was more pronounced with antibiotic therapy alone, 10.5 x 10(4) CFU per g of tibial bone were measured . However, changes were most marked using a 4-wk combination therapy consisting of HBO2 and an antibiotic agent . The colony count was 2.7 x 10(3) CFU . Each of the treatment modalities resulted in a significant therapeutic effect . The results not only demonstrated the effectiveness of HBO2 in the treatment of osteomyelitis, but revealed a potential additive effect with the combination of HBO2 and an antibiotic. Am J Rhinol, 1999 Jul-Aug, 13(4), 273 - 7 Post-functional endoscopic sinus surgery methicillin-resistant Staphylococcus aureus sinusitis; Jiang RS et al.; Methicillin-resistant Staphylococcus aureus (MRSA) is a highly virulent bacterium that is difficult to eradicate . It has become a common nosocomial pathogen, but it also causes sporadic infections in some outpatients . Among 358 chronic sinusitis patients who received functional endoscopic sinus surgery (FESS) for treatment between July 1995 and June 1997 in our department, 18 were infected postoperatively by MRSA by the end of August 1997 . One patient was excluded because she received another nasal surgery, partial turbinectomy, and submucous resection of the nasal septum, after FESS . Most of 17 MRSA infected patients presented themselves with mucopurulent nasal discharge and/or nasal crust . The treatment was generally difficult because MRSAs were resistant to multiple antibiotics . When quinolone antibiotics were used to treat most patients, the improvement rate was 76.5% . We conclude that MRSA infections in post-FESS patients might affect the outcome of FESS. Aust N Z J Ophthalmol, 1999 Jun-Aug, 27(3-4), 237 - 40 Detection of staphylococcal superantigens from contact-lens-induced inflammatory diseases; Zhu H et al.; PURPOSE: We investigated the correlation between the production of superantigens in Staphylococcus-aureus-induced ulcer and contact-lens-induced peripheral ulcer (CLPU) and infiltrative keratitis (IK) . METHODS: Twenty-five S . aureus strains used in the study were isolated either from CLPU cases (six), or IK events (13) or asymptomatic eyes (six) . A polymerase chain reaction (PCR) was employed to detect genes for superantigenic staphylococcal enterotoxins A-D, toxic shock syndrome toxin-1 and exfoliative toxins A and B . Reversed passive latex aggregation, enzyme-linked immunosorbent assay (ELISA) and immunoblot were used to examine the production of staphylococcal enterotoxins A-D and toxic shock syndrome toxin-1 . RESULTS: The frequency of the superantigenic genes was not significantly different among the clinical isolates when asymptomatic isolates were compared with the strains isolated from CLPU and IK events . ELISA and immunoblot showed non-specific protein cross-reactions . CONCLUSIONS: The results suggest that detection of superantigenic toxin genes in ocular strains of S . aureus by PCR is a specific, sensitive, rapid and inexpensive alternative to traditional immunological assays . The role, if any, of the S . aureus superantigens in the production of CLPU/IK remains uncertain. Aust N Z J Ophthalmol, 1999 Jun-Aug, 27(3-4), 224 - 7 Effect of lysozyme on adhesion and toxin release by Staphylococcus aureus; Thakur A et al.; PURPOSE: Staphylococcus aureus is a common cause of ocular infection and inflammation . We hypothesized that potential for S . aureus to cause an ocular infection would be enhanced if these bacteria are able to adhere to the biomaterials used in contact lenses . In turn, bacterial adhesion could also be influenced by other factors, such as properties of contact lenses and the absorbance of some tear components . We investigated the effect of the tear protein lysozyme on S . aureus adhesion to contact lenses and its effect on production of toxins or enzymes . METHODS: Bacterial adhesion on contact lenses was determined by counting the total number of bacteria as well as viable bacteria on lysozyme-coated or non-coated lenses, and by counting bacteria grown in the presence or absence of lysozyme in the medium . Toxin and enzyme production was assessed by haemolysis and proteolysis assays . RESULTS: Our results indicate that adhesion was significantly increased in the presence of lysozyme, both in the medium and coated onto contact lenses (P=0.04) . The presence of lysozyme did not alter the production of alpha- or beta- toxins . However, the presence of lysozyme inhibited elastase activity . CONCLUSION: These results indicate that lysozyme deposition on contact lenses promoted S . aureus adhesion . The tear protein lysozyme might modify elastase activity and thus modulate the production of corneal degradation resulting from the action of this enzyme. Surg Today, 1999, 29(8), 724 - 9 New methods of control against postoperative methicillin-resistant Staphylococcus aureus infection; Kusachi S et al.; The incidence of postoperative infections caused by methicillin-resistant Staphylococcus aureus (MRSA) in Japan has been increasing dramatically . In March 1990, we assigned special doctors in infection control (infection control doctor, ICD), and defined comprehensive controls against MRSA infection . A total of 3536 cases of digestive tract surgery performed at our department were studied during the period between September 1987 and August 1997 . We changed the use of antibiotics to prevent postoperative infection . Cefazolin (CEZ) was employed for surgery of the upper digestive tract, including esophagus, stomach, duodenum, and gallbladder . Cefotiam (CTM) was employed for surgery of the lower digestive tract, liver, and pancreas . In esophageal resection, the tracheal tube was extracted during the early postoperative period, and for cervical esophagogastroanastomosis, the autosuture was changed to layer-to-layer anastomosis . We have achieved successful control of postoperative MRSA infection, the incidence having decreased to 0.3% (9/2703) . In conclusion, our methods of control against postoperative MRSA infection implies that comprehensive measures of prevention, including the reviewed specification and usage of antibiotics and operation management, have been well implemented . This value is the lowest and the first of any domestic hospital or institute in Japan, suggesting a continued and significant decrease. Mikrobiol Z, 1999 May-Jun, 61(3), 46 - 51 {The abdominal cavity microflora of children with appendicular peritonitis}; Veselyi SV; Specific features and variability of microflora have been investigated in 75 patients with appendicular peritonitis . The patients' age was from 2 to 15 . Forty-five (45) children had local peritonitis, 17, diffuse and 13 children had total peritonitis . Complications were found in 18 (49.5%) patients . Appendicular peritonitis in children was accompanied by the permanent competition and changes in microflora . Microbiological pattern depended on the process stage and varied during a rather short period . The abundance of microorganism species during local peritonitis was gradually replaced by the prevalence of Staphylococcus aureus and especially Escherichia coli under the spread inflammation . The change of microflora during post-operation period was observed in 21 patients (56.8%) . In 9 patients the change was detected on the 15th--3rd day after the operation . Complications were observed in 85.7% of patients with changed microflora in the post-operation period . A conclusion was made that the prescription of the optimal antibacterial schemes of peritonitis treatment in children should be made with the account of change of bacterial agents during pathological process. Scand J Infect Dis, 1999, 31(3), 319 - 21 Primary psoas abscess due to Fusobacterium nucleatum; Smetana GW; A case of primary pyogenic psoas abscess due to Fusobacterium nucleatum is described . Clinicians must maintain a high index of clinical suspicion for the diagnosis of psoas abscess . Although Staphylococcus aureus accounts for most cases of primary psoas abscess, this report emphasizes the importance of bacteriological confirmation of the microorganism involved. Pneumonol Alergol Pol, 1999, 67(1-2), 60 - 4 {A case of empyema after pneumonectomy caused by methicillin-resistant staphylococcus aureus infection treated successfully with local administration with vancomycin}; Kachel T et al.; A 55-year-old man underwent left pneumonectomy due to squamous cell carcinoma . Three weeks later bronchopleural fistula and pleural empyema with MRSA infection were recognized . Treatment was based on closed pleural drainage and antibiotic therapy . Initially patient was treated with trimethoprin-sulfamethoxazole and then vancomycin intravenously but empyema was not cured completely . Therefore repeated instillation of vancomycin into the empyema cavity was applied . After 6 days of treatment, culture studies of the pleural fluid became negative and drainage tube was removed 5 days later . We suggest that local administration of vancomycin is an effective method in postpneumonectomy empyema with MRSA infection. Acta Med Port, 1999 Apr-Jun, 12(4-6), 169 - 76 {The diversity of Staphylococcus aureus strains isolated in a Lisbon hospital over a 4-year period}; Cristino JM et al.; Over a 4-year period, 2020 Staphylococcus aureus strains isolated in Santa Maria Hospital were studied, 26.3% of which were methicillin-resistant (MRSA) . The main specimens from which the strains were isolated included pus, blood and sputum/bronchial secretions . Isolation in blood cultures was the most common source among patients from medical units . Antimicrobial susceptibility studies showed that while in methicillin susceptible strains sensitivity to other antimicrobial agents (apart from penicillin resistance) was the rule, in MRSA strains there was resistance to most antibiotics . Only vancomycin was active against all strains . Phage typing showed that 75.5% of the strains were typable with phages at 100 x R.T.D . Among methicillin sensitive strains, a big diversity of phage patterns was observed, including phage groups I, II, III and V, as well as with phage association D11/95 . The large majority of MRSA strains were lysed by group III phages, although several distinct patterns were observed . Within these strains, lysis by groups II and V phages was not observed . Plasmid profiling was the least discriminant issue in the characterization of these micro-organisms because most of the strains harboured only one plasmid (or none) . These results showed that a dominant MRSA strain did not exist in this hospital, but rather several distinct strains . The importance, as well as the difficulties in controlling the spread of MRSA strains in the present conditions of high prevalence, are highlighted. Microbiol Immunol, 1999, 43(6), 505 - 12 Quantification of phagocytosis in human neutrophils by flow cytometry; Heinzelmann M et al.; Phagocytosis represents a central element of the host response to microbial invasion . We describe a flow cytometric method for measuring the kinetics of phagocytosis of two bacteria by human polymorphonuclear leukocytes (PMNs) . Over a 60-min period, isolated human PMNs were exposed to Staphylococcus aureus (rapidly phagocytosed) and Klebsiella pneumoniae (slowly phagocytosed) . This method distinguished adherent from ingested bacteria by quenching fluorescein isothiocyanate-labeled extracellular bacteria with ethidium bromide . This further allowed the exclusion of dead, highly permeable, and subsequently bright-red fluorescent PMNs . Our experiments with two different bacteria, various PMN-to-bacteria ratios (1:1, 1:10, 1:100), and different individuals proved that 1) flow cytometric analysis is accurate and useful for characterizing phagocytosis, 2) adherent bacteria can be distinguished from ingested bacteria after quenching with ethidium bromide, and that 3) phagocytosis kinetics of two bacteria with different onsets of phagocytosis can be determined by flow cytometry and the assessment of a score that quantifies phagocytosis. J Infect Dis, 1999 Oct, 180(4), 1370 - 3 Protection against Staphylococcus aureus sepsis by vaccination with recombinant staphylococcal enterotoxin A devoid of superantigenicity; Nilsson IM et al.; Staphylococcal exotoxins are virulence determinants in Staphylococcus aureus arthritis and septicemia . To assess the utility of enterotoxins as vaccine candidates for these diseases, a genetically modified staphylococcal enterotoxin A (SEA) that lacks superantigenic properties was used . Mice immunized with recombinant (r) SEA had significantly longer survival than control immunized mice and lost significantly less weight than the controls . Transfer of SEA-specific antibodies to naive mice resulted in good protection against death in staphylococcal sepsis . In vitro proliferative responses to SEA by naive lymphocytes were almost totally abolished on incubation with serum from rSEA but not with control antigen-immunized mice . These results suggest that immunization with rSEA devoid of superantigenic properties provides good protection against S . aureus sepsis . In addition, the data indicate that the protection is at least in part mediated by SEA neutralizing antibodies. J Infect Dis, 1999 Oct, 180(4), 1365 - 9 Cross-reactive antibodies prevent the lethal effects of Staphylococcus aureus superantigens; Bavari S et al.; The exotoxins produced by Staphylococcus aureus, staphylococcal enterotoxins (SE) A-E and toxic shock syndrome toxin (TSST)-1, which are associated with serious diseases, including food poisoning and toxic shock syndrome, are termed superantigens (SAgs) . To examine whether common antigenic epitopes were present and whether vaccination with 1 bacterial SAg could protect against challenge with a different SE or TSST-1, mice were vaccinated with SEA, SEB, SEC1, or TSST-1 individually or in combination . Mice injected with a single toxin developed high antibody titers against other SAgs . Marked improvement in survival was observed when immunized mice were challenged with a heterologous toxin . Mice vaccinated with a mixture of toxins were fully protected against 1 or multiple toxin challenges, indicating no interference effects of multivalent vaccinations . More importantly, higher titers were found against each SAg with the multivalent vaccination than with injection with a single SAg . Thus, immunizations with 1 SAg can induce cross-protective antibodies to heterologous SAgs, and multicomponent vaccination can enhance antibody responses against each bacterial SAg. Cell Transplant, 1999 Jul-Aug, 8(4), 405 - 11 Effectiveness of acidic oxidative potential water in preventing bacterial infection in islet transplantation; Miyamoto M et al.; At a number of points in the current procedures of islet isolation and islet culture after the harvesting of donor pancreata, microorganisms could potentially infect the islet preparation . Furthermore, the use of islets from multiple donors can compound the risks of contamination of individual recipients . Acidic oxidative potential water (also termed electrolyzed strong acid solution, function water, or acqua oxidation water), which was developed in Japan, is a strong acid formed on the anode in the electrolysis of water containing a small amount of sodium chloride . It has these physical properties: pH, from 2.3 to 2.7; oxidative-reduction potential, from 1,000 to 1,100 mV; dissolved chlorine, from 30 to 40 ppm; and dissolved oxygen, from 10 to 30 ppm . Because of these properties, acidic oxidative potential water has strong bactericidal effects on all bacteria including methicillin-resistant Staphylococcus aureus (MRSA), viruses including HIV, HBV, HCV, CMV, and fungi as a result of the action of the active oxygen and active chlorine that it contains . We conducted this study to evaluate the effect of acidic oxidative potential water irrigation on bacterial contamination on the harvesting of porcine pancreata from slaughterhouses for islet xenotransplantation by counting the number of pancreatic surface bacteria using the Dip-slide method, and on the results of islet culture; and to evaluate the direct effect on isolated islets when it is used to prevent bacterial contamination by the static incubation test and by morphological examination . Direct irrigation of the pancreas by acidic oxidative potential water was found to be very effective in preventing bacterial contamination, but direct irrigation of isolated islets slightly decreased their viability and function. Radiology, 1999 Sep, 212(3), 687 - 92 Osteoid osteoma: CT-guided percutaneous resection and follow-up in 38 patients; Sans N et al.; PURPOSE: To reevaluate at medium term the results of computed tomography (CT)-guided percutaneous resection of osteoid osteomas . MATERIALS AND METHODS: Thirty-eight patients who had undergone treatment by means of this technique were reexamined with a mean follow-up of 3.7 years . The short- and medium-term clinical course and histologic features of the resection specimens were analyzed . RESULTS: The bone fragment could be analyzed in all cases, and the diagnosis of osteoid osteoma was confirmed in 28 patients (74%) . A different diagnosis was made in six patients: mucoid cyst, subchondral arthritic geode, fibrous dysplastic lesion, focal osteochondritis, or focal chronic osteomyelitis . Cure was obtained in 32 patients (84%), whatever the cause . Complications, generally minor and transient, were observed in nine patients (24%) . The most severe complications were two femoral fractures and one focal chronic osteomyelitis due to Staphylococcus aureus infection . CONCLUSION: The results of this study confirm the efficacy of percutaneous resection of osteoid osteomas and the possibility of using this method for successful treatment of other small bone lesions. Radiol Med (Torino), 1999 Jun, 97(6), 467 - 71 {Chronic spondylodiscitis . Clinical aspects and imaging features}; Meneghello A et al.; INTRODUCTION: The diagnosis of a chronic inflammatory process involving the vertebral body and disk is often very difficult because patient's history, subjective symptoms and physical findings are often unconclusive . Thus imaging techniques play a decisive role . Radiography, tomography, CT and MR have different capabilities and limitations and provide different findings in spondylodiscitis . MATERIAL AND METHODS: We observed 18 cases of spondylodiscitis in the last three years . The responsible microbe, a Staphylococcus aureus from extraosseous sites, was found in two cases at blood culture . Small cell inflammatory infiltration was confirmed with CT-guided biopsy in one case, while the other cases were diagnosed based on constant chronic back pain, feveret, moderate neutrophile leukocytosis or increased erythrosedimentation speed, plus changes in radiographic patterns following antibiotic therapy . RESULTS: Plain radiography and tomography are the techniques of choice to detect or suspect the lesion, which is then studied with CT or MRI . Clear-cut irregularities and erosions on opposing vertebral bodies, reactive bone sclerosis and reduced disk space were typical signs in our series; nine patients presented irregular cavitations(s), like bone caries, surrounded by reactive sclerosis in the body near the frontal vertebral plate . CONCLUSIONS: Together with the imaging patterns of all cases, we studied in detail three cases, relative to physical findings and diagnostic techniques . We also compared the changes in chronic spondylodiscitis with those in intraspongious herniation, intervertebral osteochondritis and severe degenerative arthritis . Bone erosions on the anterior cortical surface of the vertebral body were seen in 50% of our cases and may represent a specific sign of chronic spondylodiscitis if the finding is confirmed in further studies. Lik Sprava, 1999 Apr-May, (3), 79 - 82 {The development of a coagglutination reaction using staphylococcal peptidoglycan for the diagnosis of staphylococcal infection}; Pozur VK et al.; Our objective in this study was to develop and assess the diagnostic value of a coagglutination test with making use of peptidoglycane of Staphylococcus aureus in the identification of diseases of staphylococcal etiology . A total of 166 patients with diseases of staphylococcal etiology were examined . A test was elaborated of coagglutination with making use of peptidoglycane of Staphylococcus aureus for a differential identification of antibodies to peptidoglycan in healthy persons and patients with staphylococcosis. Lik Sprava, 1999 Apr-May, (3), 42 - 4 {The effect of ecological and microbiological factors on the health status of pregnant women with pyelonephritis and on their newborn infants}; Iashchukevych MIe; An analysis was performed of history, course of pregnancy, parturition, and condition of the newborn babies in female patients with pyelonephritis depending on the microbiological factor and environmental situation in the zone of residence . The pregnant women living under adverse environmental conditions display high levels of endocrine, cardiovascular, gastrointestinal disorders, and of tonsillitis . In the majority of cases, pregnancy is noted to be complicated by anemia (76.7%) and fetoplacental incompetence (62.9%), with infants being born in asphyxia presenting with signs of hypotrophy, congenital infection . Of the above infants, 37% develop postnatal inflammatory conditions . Two variants were shown to play a part in the etiology of pyelonephritis: monoetiological one marked by predominance of Staphylococcus aureus and Escherichia coli and polyetiological variant characterized by predominance of Candida fungi, Staphylococcus aureus, and mycoplasma . Irrespective of the microbiological factor, the female patients with pyelonephritis demonstrated high levels of premature birth, had a history of infertility, and were presenting with genital and extragenital pathologies. Nucleosides Nucleotides, 1999 Jun-Jul, 18(6-7), 1297 - 9 Detection of nucleic acids by cycling probe technology on magnetic particles: high sensitivity and ease of separation; Bhatt R et al.; Cycling Probe Technology (CPT) is a signal amplification system that allows detection of nucleic acid target sequences without target amplification . CPT employs a sequence specific chimeric probe, typically DNA-RNA-DNA, which hybridizes to a complementary target DNA sequence and becomes a substrate for RNase H . Cleavage occurs at the RNA internucleotide linkages and results in dissociation of the probe from the target, thereby making it available for the next probe molecule . This communication describes the use of oligonucleotides attached to solid supports for target capture and release followed by solution and solid phase cycling . Through the attachment of chimeric probes to Sera-Mag magnetic particles (SMP) a simple and effective method of separating the cleaved probe from non-cycled probe has been developed . By capturing the target DNA on particles and separating it from the extraneous non-specific DNA we are able to dramatically reduce background and thus discriminate between samples of Methicillin Resistant (MRSA) and Methicillin Sensitive (MSSA) Staphylococcus Aureus . We conjugated oligonucleotide probes to SMPs (approximately 1 um) and Nylon beads (NB) which were coated with ID Biomedical's proprietary coating materials (R, patent pending) . The general structure of the constructs is shown below: {table: see text} Ophthalmic Res, 1999, 31(6), 446 - 51 Ofloxacin levels after intravitreal injection . Effects of trauma and inflammation; Ozturk F et al.; PURPOSE: This study was carried out to get an insight into the ofloxacin elimination after intravitreal injection in rabbits . We also studied the effects of trauma and inflammation on the vitreous ofloxacin levels after intravitreal injection of ofloxacin . METHODS: A penetrating eye injury in the right eye was inflicted on 24 rabbits and another 12 animals were used as control . A standardized intraocular inflammation was induced by intravitreal injection of a suspension of Staphylococcus aureus in half of the traumatized eyes . Ofloxacin (200 microg/0.1 ml) was injected into the midvitreous cavity of both traumatized and control right eyes, and samples were obtained at 2, 8, 24 and 48 h after injection . Drug concentrations were measured using high-pressure liquid chromatography analysis . RESULTS: Vitreous levels of ofloxacin were above the MIC(90) at 2 and 8 h in all groups for most of the common microorganisms causing endophthalmitis and also at 24 h in traumatized-infected eyes . At the second hour, the mean vitreous concentrations of ofloxacin both in traumatized and traumatized-infected eyes were lower than that in the control eyes (p < 0.05) . At 8 h, the mean vitreous concentrations of ofloxacin in the traumatized and in the traumatized-infected eyes were higher than that in the control eyes (p < 0.05) . At 24 h, the mean ofloxacin concentration was higher in the traumatized-infected eyes than that in control (p < 0.01) and traumatized eyes (p < 0.05), and also higher in the traumatized eyes than that in the control eyes (p < 0.05) . The mean ofloxacin concentrations in the traumatized and traumatized-infected eyes were significantly higher (p < 0.01) than those in the controls at 48 h . The elimination half-life of ofloxacin in the control eyes was 5.65 h and trauma and inflammation prolonged the half-life to 9.47 and 9 . 72 h, respectively . CONCLUSION: Clearance of ofloxacin is fast and appears to be reduced by trauma and inflammation . Therapeutic drug levels in traumatized-infected eyes were maintained up to 24 h . This may be an important pharmacokinetic advantage in treating endophthalmitis unless the dose used has local toxicity and allows a longer dose interval when the dose is repeated. J Antimicrob Chemother, 1999 Aug, 44(2), 263 - 73 Treatment of hospitalized patients with complicated gram-positive skin and skin structure infections: two randomized, multicentre studies of quinupristin/dalfopristin versus cefazolin, oxacillin or vancomycin . Synercid Skin and Skin Structure Infection Group; Nichols RL et al.; Quinupristin/dalfopristin (Synercid), the first injectable streptogramin antibiotic available for the treatment of complicated gram-positive skin and skin structure infections, was compared with standard comparators (cefazolin, oxacillin or vancomycin) in one USA and one international trial . These two randomized, open-label trials of virtually identical design enrolled a total of 893 patients (450 quinupristin/dalfopristin, 443 comparator) . The majority of patients had erysipelas, traumatic wound infection or clean surgical wound infection . Staphylococcus aureus was the most frequently isolated pathogen in both treatment groups and polymicrobial infection was more common in the quinupristin/dalfopristin group than in the comparator group . The clinical success rate (cure plus improvement) in the clinically evaluable population was equivalent between the two treatment groups (68.2% quinupristin/dalfopristin, 70.7% comparator; 95% CI, -10.1, 5.1) despite a shorter mean duration of treatment for quinupristin/dalfopristin patients . In the bacteriologically evaluable population, by-patient and by-pathogen bacteriological eradication rates were somewhat lower for quinupristin/dalfopristin (65.8% and 66.6%, respectively) than for the comparator regimens (72.7% and 77.7%, respectively) . The lower bacteriological response rates in the quinupristin/dalfopristin group were, in part, due to a higher rate of polymicrobial infections and a higher incidence of patients classified as clinical failure, a category which included premature discontinuation of treatment because of local venous adverse events . The bacteriological eradication rate for quinupristin/dalfopristin was higher in monomicrobial infections than in polymicrobial infections (72.6% versus 63.3%, respectively), whereas the corresponding rate for the comparator regimens was lower for monomicrobial infections than polymicrobial infections (70.8% versus 83.1%) . This finding was not unexpected, since the spectrum of quinupristin/dalfopristin is focused on gram-positive pathogens and additional antibiotics to treat gram-negative bacteria were not required per protocol . The systemic tolerability of both treatment regimens was qualitatively similar . A higher rate of drug-related venous adverse events was reported for quinupristin/dalfopristin (66.2%) than for the comparator regimen (28.4%) . Premature discontinuation of study drug was primarily due to adverse clinical events for quinupristin/dalfopristin (19.1%), whereas the most common reason for discontinuation among those receiving the comparator regimens was treatment failure (11.5%) . Quinupristin/dalfopristin is an effective alternative for the treatment of hospitalized patients with complicated skin and skin structure infections due to quinupristin/ dalfopristin-susceptible gram-positive organisms, including methicillin- and erythromycin-resistant S . aureus. J R Soc Med, 1999 Jun, 92(6), 283 - 5 Antibacterial activity of honey against strains of Staphylococcus aureus from infected wounds; Cooper RA et al.; The antibacterial action of honey in infected wounds does not depend wholly on its high osmolarity . We tested the sensitivity of 58 strains of coagulase-positive Staphylococcus aureus, isolated from infected wounds, to a pasture honey and a manuka honey . There was little variation between the isolates in their sensitivity to honey: minimum inhibitory concentrations were all between 2 and 3% (v/v) for the manuka honey and between 3 and 4% for the pasture honey . Thus, these honeys would prevent growth of S . aureus if diluted by body fluids a further seven-fold to fourteen-fold beyond the point where their osmolarity ceased to be completely inhibitory . The antibacterial action of the pasture honey relied on release of hydrogen peroxide, which in vivo might be reduced by catalase activity in tissues or blood . The action of manuka honey stems partly from a phytochemical component, so this type of honey might be more effective in vivo . Comparative clinical trials with standardized honeys are needed. Appl Environ Microbiol, 1999 Sep, 65(9), 4134 - 40 Staphylococcal surface display of immunoglobulin A (IgA)- and IgE-specific in vitro-selected binding proteins (affibodies) based on Staphylococcus aureus protein A; Gunneriusson E et al.; An expression system designed for cell surface display of hybrid proteins on Staphylococcus carnosus has been evaluated for the display of Staphylococcus aureus protein A (SpA) domains, normally binding to immunoglobulin G (IgG) Fc but here engineered by combinatorial protein chemistry to yield SpA domains, denoted affibodies, with new binding specificities . Such affibodies, with human IgA or IgE binding activity, have previously been selected from a phage library, based on an SpA domain . In this study, these affibodies have been genetically introduced in monomeric or dimeric forms into chimeric proteins expressed on the surface of S . carnosus by using translocation signals from a Staphylococcus hyicus lipase construct together with surface-anchoring regions of SpA . The recombinant surface proteins, containing the IgA- or IgE-specific affibodies, were demonstrated to be expressed as full-length proteins, localized and properly exposed at the cell surface of S . carnosus . Furthermore, these chimeric receptors were found to be functional, since recombinant S . carnosus cells were shown to have gained IgA and IgE binding capacity, respectively . In addition, a positive effect in terms of IgA and IgE reactivity was observed when dimeric versions of the affibodies were present . Potential applications for recombinant bacteria with redirected binding specificity in their surface proteins are discussed. J Antimicrob Chemother, 1999 Aug, 44(2), 193 - 9 Intracellular activity of trovafloxacin against Staphylococcus aureus; van den Broek PJ et al.; The effect of trovafloxacin on Staphylococcus aureus ingested by human granulocytes or monocytes was compared with that on S . aureus in cell-free medium . Maximum growth inhibition (E(R,max)) by the antibiotic was 0.530 log10/h for S . aureus within granulocytes, 0.912 log10/h for S . aureus within monocytes, and 1.830-1.916 log10/h for S . aureus in medium . EC50, the concentration at which 50% of the maximum growth inhibition is achieved, did not differ significantly under the conditions investigated . After inhibition of intracellular killing by granulocytes with sodium fluoride, the intracellular antibacterial activity of trovafloxacin was still less than that in medium . A 3.4 times higher concentration was needed to achieve the same effect on phagocytosed S . aureus as in cell-free medium . Trovafloxacin binds more strongly to granulocytes than to monocytes, the respective cellular concentrations being 10 and four times higher than that in medium . In conclusion, the activity of trovafloxacin against S . aureus ingested by human granulocytes or monocytes is less than that against S . aureus in cell-free medium and is not related to the cell-associated concent