Rinsho Shinkeigaku, 2000 Jan, 40(1), 8 - 13 {MRSA infection control in the wards for progressive muscular dystrophy: the effects of encouraged handwashing}; Matsumura T et al.; There are currently 27 national hospitals with the wards for progressive muscular dystrophy (PMD) in Japan . Today, most patients in these wards are severely motor handicapped and complicated with respiratory and/or cardiac failure . Malnutrition, dysphagia, insufficient respiratory tract clearance are common problems that cause fragility to infection . Although progress in the treatment of infection have remarkably prolonged their life-span, frequent use of antibiotics is a major factor for occurrence of drug resistant bacteria . Since we had the first case of methicillin resistant Staphylococcus aureus (MRSA) infection in 1994 in our hospital, the number of MRSA carriers increased year by year . To prevent the spread of MRSA, we revised our infection precaution manual and surveyed its consequence . We encouraged handwashing of staffs, introduced green tea in cleansing genital region and abolished the use of a private room except a patient with severe seborrheic eczema . The revision of the manual resulted in decrease of MRSA carriers . The surveillance revealed that many patients had MRSA in genital regions, although there were no relationships between colonization of MRSA and severlity of disability or complications . It was strongly suggested that toilettary care by the hands of nursing staffs was a major factor of transferring MRSA . Our study revealed that encouragement of handwashing is more powerful in preventing the spreading MRSA, and more favorable for quality of lives of PMD patients than isolating the patients. J Gastroenterol Hepatol, 2000 Apr, 15(4), 431 - 6 Decreased function of peripheral blood dendritic cells in patients with hepatocellular carcinoma with hepatitis B and C virus infection; Kakumu S et al.; BACKGROUND: Tumour immunity does not seem to be induced effectively in tumour-bearing hosts, including in patients with hepatocellular carcinoma (HCC) . One possible reason is that function of dendritic cells (DC) is decreased in such hosts . METHODS: We evaluated T cell stimulatory activity and interleukin (IL)-12 production of DC and interferon (IFN)-gamma and IL-10 production of T cells of peripheral blood from 12 control individuals and 21 patients with chronic hepatitis C virus (HCV) infection (six with chronic hepatitis (CH), eight with liver cirrhosis (LC) and 13 with HCC) . Five hepatitis B virus (HBV)-infected patients with HCC were included as a disease control group . The DC were prepared by the culture of T cell-depleted populations of peripheral blood mononuclear cells in the presence of granulocyte-macrophage colony stimulating factor and IL-4 for a total of 11-12 days . The cytokine levels were assayed by ELISA . To test the stimulatory function of DC in T cell proliferation, mytomycin C-treated DC were cultured with allogeneic T cells from a control . RESULTS: When the T cell-stimulatory activity of DC was expressed as stimulation index value of {3H}-thymidine incorporation of T cells, the values were lower in HCV-infected HCC (2.6 +/- 1.8, P < 0.01) than in controls (5.5 +/- 2.0) and CH (5.0 +/- 1.3) . Staphylococcus aureus Cowan 1-induced IL-12 production of DC was decreased in HCV-infected HCC (P < 0.001, P < 0.01 and P < 0.05, respectively) compared with controls, CH and LC, while similar amounts of IL-10 were produced in patients and controls . Interleukin-10 and IFN-gamma production of T cells in response to anti-CD3 antibody or IL-12 were equivalent between patient groups and controls, respectively . Similarly decreased DC function and normal T cell response were observed in HBV-infected HCC patients . CONCLUSIONS: These findings suggest that the depressed function of DC is associated with pathogenesis of HCC with HBV or HCV infection. J Infect Dis, 2000 May, 181(5), 1693 - 9 Epub 2000 May 15. Induction of acute pleural inflammation by Staphylococcus aureus . I . CD4+ T cells play a critical role in experimental empyema; Mohammed KA et al.; Bacterial empyema is a frequent complication of pneumonia in patients with acquired immunodeficiency syndrome (AIDS) . A model of Staphylococcus aureus empyema was developed that closely resembles bacterial empyema in patients infected with human immunodeficiency virus (HIV) . Results show a compartmentalized chemokine response in bacterial empyema . The chemokine levels were higher in the pleural compartment than in the peripheral circulation . Polymorphonuclear leukocyte counts, murine GRO-alpha (KC), and macrophage inflammatory protein-2 levels were significantly (P<.001) lower in CD4+ knockout (CD4 KO) mice pleural fluid than in CD4+ wild-type (CD4 WT) mice . The CD4 KO mice had poorer bacterial clearance than CD4 WT mice . During S . aureus infection, interleukin-10 levels increased in the CD4 KO mice, whereas interferon-gamma levels were increased in CD4 WT mice . CD4+ T cell depletion results in a decreased pleural chemokine response, decreased neutrophil influx into pleural space, and impaired bacterial clearance in empyema. Infect Control Hosp Epidemiol, 2000 May, 21(5), 319 - 23 Nasal carriage of Staphylococcus aureus is a major risk factor for surgical-site infections in orthopedic surgery; Kalmeijer MD et al.; OBJECTIVE: To determine the relative importance of different risk factors for the development of surgical-site infections (SSIs) in orthopedic surgery with prosthetic implants . DESIGN: In a cohort of 272 patients, the following possible risk factors were studied: age, gender, method of hair removal, duration of operation, surgeon, underlying illness, and nasal carriage of Staphylococcus aureus . Infections were recorded following the Centers for Disease Control criteria . The relation between risk factors and SSI was tested in univariate and multiple logistic regression analysis . SETTING: Community hospital in Breda, The Netherlands . RESULTS: 18 (6.6%) of 272 patients experienced SSI: 11 superficial and 7 deep SSI . These infections led in three cases to removal of the prosthesis and caused 286 extra days in hospital . The main causative pathogen was S aureus . In multiple logistic regression analysis, the following factors were independent risk factors for the development of SSI: high-level nasal carriage of S aureus (P=.04), male gender (P=.005), and surgeon 1 (P=.006) . The only independent risk factor for SSI with S aureus was high-level nasal carriage of S aureus (P=.002) . CONCLUSION: High-level nasal carriage of S aureus was the most important and only significant independent risk factor for developing SSI with S aureus. Infect Control Hosp Epidemiol, 2000 May, 21(5), 311 - 8 An operating surveillance system of surgical-site infections in The Netherlands: results of the PREZIES national surveillance network . Preventie van Ziekenhuisinfecties door Surveillance; Geubbels EL et al.; OBJECTIVES: To describe the results of the first year of the Dutch national surveillance of surgical-site infections (SSIs) and risk factors, which aims to implement a standardized surveillance system in a network of Dutch hospitals, to collect comparable data on SSIs to serve as a reference, and to provide a basic infrastructure for further intervention research . DESIGN: Prospective multicenter cohort study . SETTING: Acute-care hospitals in The Netherlands from June 1996 to May 1997 . RESULTS: 38 hospitals participated, with a slight over-representation of larger hospitals . Following a total of 18,063 operations, 562 SSIs occurred, of which 198 were deep . Multivariate analysis of pooled procedures shows that age, preoperative length of stay, wound contamination class, anesthesia score, and duration of surgery were independent risk factors for SSI . When analyzed by procedure, the relative importance of these risk factors changed . Bacteriological documentation was available for 56% of the SSIs; 35% of all isolates were Staphylococcus aureus . Multiple regression analysis computed the mean extra postoperative length of stay associated with SSI to be 8.2 days . CONCLUSION: The first year of national surveillance has shown that it is feasible to collect comparable data on SSI, which are already used for education, policy, and decision making in the network of participating hospitals . This gives room to effectuate the next aim, namely to use the network as an infrastructure for intervention research . Multivariate analysis shows that feedback on a procedure-specific level is important. J Trauma, 2000 May, 48(5), 918 - 23 Association of endotoxemia and production of antibodies against endotoxins after multiple injuries; Buttenschoen K et al.; BACKGROUND: Endotoxemia after injury has been a controversial issue . Endotoxins stimulate the innate and adaptive immune system . OBJECTIVE: To investigate endotoxemia and its effects on the production of antiendotoxin antibodies of cultured mononuclear cells of patients with multiple injuries . METHODS: Blood samples of 20 patients with multiple injuries were collected up to 12 days after trauma . The endotoxin concentration was measured in the plasma, and mononuclear cells were isolated and cultured . Specific antibodies against two lipopolysaccharides, one lipid A preparation, and alpha-hemolysin of Staphylococcus aureus were measured in the cell culture supernatant by an enzyme-linked immunosorbent assay . RESULTS: Endotoxemia peaked at admission of the patients, decreasing thereafter to almost normal values within 5 days . Isolated mononuclear cells synthesized antibodies against all tested antigens with a peak at or between day 5 and day 7 . The increase was significant for immunoglobulin (Ig)A and IgM specific to all endotoxins tested and for IgA specific to alpha-hemolysin . However, there were no significant changes of the concentrations of total IgM, IgA, and IgG . All specific IgG remained unaffected . CONCLUSION: Patients with multiple injuries initially have temporary endotoxemia . Endotoxin may be suggested as a stimulator of the synthesis of antiendotoxin antibodies, in particular of the IgA and IgM class in patients with multiple injuries. Thromb Haemost, 2000 May, 83(5), 777 - 84 Mechanisms involved in the antiplatelet activity of Staphylococcus aureus lipoteichoic acid in human platelets; Sheu JR et al.; In this study, gram-positive Staphylococcus aureus lipoteichoic acid (LTA) dose-dependently (0.1-1.0 microg/ml) and time-dependently (10-60 min) inhibited platelet aggregation in human platelets stimulated by agonists . LTA also dose-dependently inhibited phosphoinositide breakdown and intracellular Ca+2 mobilization in human platelets stimulated by collagen . LTA (0.5 and 1.0 microg/ml) also significantly inhibited thromboxane A2 formation stimulated by collagen in human platelets . Moreover, LTA (0.1-1.0 microg/ml) dose-dependently decreased the fluorescence of platelet membranes tagged with diphenylhexatrience . Rapid phosphorylation of a platelet protein of Mr . 47,000 (P47), a marker of protein kinase C activation, was triggered by PDBu (30 nM) . This phosphorylation was markedly inhibited by LTA (0.5 and 1.0 microg/ml) within a 10-min incubation period . These results indicate that the antiplatelet activity of LTA may be involved in the following pathways: LTA's effects may initially be due to induction of conformational changes in the platelet membrane, leading to a change in the activity of phospholipase C, and subsequent inhibition of phosphoinositide breakdown and thromboxane A2 formation, thereby leading to inhibition of both intracellular Ca+2 mobilization and phosphorylation of P47 protein . Therefore, LTA-mediated alteration of platelet function may contribute to bleeding diathesis in gram-positive septicemic and endotoxemic patients. J Qual Clin Pract, 2000 Mar, 20(1), 6 - 11 Standardising surveillance of nosocomial infections: the HISS program . Hospital Infection Standardised Surveillance; McLaws ML et al.; Standardised surveillance of nosocomial infections in Australia had not been addressed until June 1998 when the New South Wales Health Department funded the development and implementation of the first standardised surveillance system for hospital infection: the Hospital Infection Standardised Surveillance program (HISS) . The introduction of a standardised surveillance system needs to balance the requirements of a Health Department and the needs of hospitals . The Health Department requires data to develop aggregated rates for the setting of thresholds for all nosocomial infections while hospitals require rates to reflect the quality of clinical care and provide data for evidence-based infection control practices . The Hospital Infection Epidemiology and Surveillance (HIES) Unit has attempted to balance these requirements using a 'sentinel surveillance' approach with standardised definitions and methodology . The HISS program utilizes eICAT software modified for its standardised requirements of data collection . To date, 10 hospitals surveyed sentinel multiple resistant organisms (MRO), eight also elected sentinel surgical procedures (SSP) and intravascular device-related bacteraemia (IVDRB) modules, and two the seasonal respiratory syncytial (RSV) and rota-virus modules in paediatric patients . The surgical site infection rates in three commonly monitored SSP were 1.8% (95% confidence interval (CI) 0.7-3.9%) for coronary artery bypass (CABG), 3.3% (95% CI 1.4-6.8%) lower segment Caesarean section (LSCS) and 7.7% (95% CI 3.4-14.6%) colorectal surgery . The rate of IVDRB was 4.7 per 1000 central venous catheter days (95% CI 2.2-8.6) and 1.1 per 1000 peripheral line-days (95% CI 0.1-3.9) . Methicillin resistant Staphylococcus aureus (MRSA) accounted for 99% of all new infections diagnosed with an endemic MRO. Ned Tijdschr Geneeskd, 2000 May 6, 144(19), 887 - 9 {Methicillin-resistant Staphylococcus aureus in medical and paramedical personnel returned from work in a foreign hospital}; Vriens MR et al.; The cultures of two patients of the Surgical Intensive Care Unit (IC) of the Medical Centre of Utrecht University were found positive for methicillin-resistant Staphylococcus aureus (MRSA) . A male nurse turned out to be the source, 4 months after his return from working in an English hospital . Cultures were, by mistake, not taken directly on arrival from abroad . Pulsed-field gel electrophoresis proved MRSA strains from both source and the 2 patients to be identical to a strain which was epidemic in Great Britain but had never occurred in the Netherlands . The IC has meanwhile been closed; at source investigation, 14 other patients and six staff members were found MRSA-positive . The policy in the hospital is to screen health care workers for MRSA carriership on return from an hospital abroad . The success of the policy depends strongly on the cooperation of health care workers in this matter. Acta Crystallogr D Biol Crystallogr, 2000 Jun, 56 ( Pt 6), 705 - 13 Ab initio structure determination of the lantibiotic mersacidin; Schneider TR et al.; The crystal structure of mersacidin, a potential novel antibiotic against methicillin- and vancomycin-resistant Staphylococcus aureus strains, has been determined by ab initio methods . Despite all crystals being merohedrally twinned, an accurate structural model with an R value of 13.4% has been obtained at atomic resolution . With six molecules in the asymmetric unit and no atom heavier than sulfur, the structure corresponds to a protein of 120 amino acids and is the largest approximately equal-atom unknown structure solved by direct methods . In the crystal, the molecule assumes a compact fold different from that found by NMR in solution . Comparison of the NCS-related molecules reveals regions of variable flexibility . The region highly homologous to the related antibiotic actagardine is very rigid and possibly defines an essential building block of this class of new antibacterial substances. Antimicrob Agents Chemother, 2000 Jun, 44(6), 1616 - 23 Phenotypic expression of oxacillin resistance in Staphylococcus epidermidis: roles of mecA transcriptional regulation and resistant-subpopulation selection; Dickinson TM et al.; The MICs for many oxacillin-resistant (OR) Staphylococcus epidermidis (ORSE) strains are below the Staphylococcus aureus methicillin or oxacillin resistance breakpoint . The difficulty detecting the OR phenotype in S . epidermidis may be due to extreme heterotypy in resistance expression and/or transcriptional repression of mecA, the OR gene, by MecI . To determine the role of these factors in the phenotypic expression of ORSE, 17 geographically diverse mecI(+) ORSE isolates representing 14 distinct pulsed-field gel electrophoresis pulse types (>3 band differences) were investigated . Thirteen of the 14 types contained mecI and mecA promoter-operator sequences known to be associated with maximal mecA repression, and in all isolates, mecA transcription was repressed . All 17 were heterotypic in their resistance expression . Oxacillin MICs ranged from 1 to 128 microg/ml and increased for 16 of 17 isolates after beta-lactam induction . Allelic replacement inactivation of mecI in three isolates similarly resulted in a four- to sevenfold increase in MIC . In the two of these three isolates producing beta-lactamase, mecA transcription was regulated by both mecI and beta-lactamase regulatory sequences . Heterotypic expression of resistance in these three isolates was unaffected by either beta-lactam induction or mecI inactivation . However, prolonged incubation in concentrations of oxacillin just sufficient to produce a lag in growth (0.5 to 1.0 microg/ml) converted the population resistance expression from heterotypic to homotypic . Homotypic conversion could also be demonstrated in microtiter wells during MIC determinations in one isolate for which the MIC was high . We conclude that the phenotypic expression of S . epidermidis OR in broth can be affected both by mecA transcriptional regulation and by subpopulation resistance expression. Antimicrob Agents Chemother, 2000 Jun, 44(6), 1549 - 55 A new class of genetic element, staphylococcus cassette chromosome mec, encodes methicillin resistance in Staphylococcus aureus; Katayama Y et al.; We have previously shown that the methicillin-resistance gene mecA of Staphylococcus aureus strain N315 is localized within a large (52-kb) DNA cassette (designated the staphylococcal cassette chromosome mec {SCCmec}) inserted in the chromosome . By sequence determination of the entire DNA, we identified two novel genes (designated cassette chromosome recombinase genes {ccrA and ccrB}) encoding polypeptides having a partial homology to recombinases of the invertase/resolvase family . The open reading frames were found to catalyze precise excision of the SCCmec from the methicillin-resistant S . aureus chromosome and site-specific as well as orientation-specific integration of the SCCmec into the S . aureus chromosome when introduced into the cells as a recombinant multicopy plasmid . We propose that SCCmec driven by a novel set of recombinases represents a new family of staphylococcal genomic elements. Antimicrob Agents Chemother, 2000 Jun, 44(6), 1428 - 37 Induction of fibronectin-binding proteins and increased adhesion of quinolone-resistant Staphylococcus aureus by subinhibitory levels of ciprofloxacin; Bisognano C et al.; We recently reported that strain EN1252a, a fluoroquinolone-resistant derivative of Staphylococcus aureus NCTC8325 with mutations in grlA and gyrA, expressed increased levels of fibronectin-binding proteins (FnBPs) and showed a significantly higher attachment to fibronectin-coated polymer surfaces after growth in the presence of subinhibitory concentrations of ciprofloxacin . The present study evaluated the occurrence and frequency of fluoroquinolone-induced FnBP-mediated adhesion in clinical isolates of fluoroquinolone-resistant methicillin-resistant S . aureus (MRSA) and methicillin-susceptible S . aureus (MSSA) . Eight of ten MRSA isolates and four of six MSSA isolates with grlA and gyrA mutations exhibited significant increases in attachment to fibronectin-coated surfaces after growth in the presence of one-quarter the MIC of ciprofloxacin . Fluoroquinolone-induced FnBP-mediated adhesion of one clinical MRSA strain and the double mutant strain EN1252a also occurred on coverslips removed from the subcutaneous space of guinea pigs . For strain EN1252a, the regulation of fnb transcription by sub-MICs of ciprofloxacin was studied on reporter plasmids carrying fnb-luxAB fusions . One-quarter of the MIC of ciprofloxacin significantly increased fnbB, but not fnbA, promoter activity of the fluoroquinolone-resistant mutant but not its fluoroquinolone-susceptible parent ISP794 . This response was abolished by pretreatment with rifampin, indicating an effect at the level of transcription . Activation of the fnbB promoter was not due to an indirect effect of ciprofloxacin on growth rate and still occurred in an agr mutant of strain EN1252a . These data suggest that sub-MIC levels of ciprofloxacin activate the fnbB promoter of some laboratory and clinical isolates, thus contributing to increased production of FnBP(s) and leading to higher levels of bacterial attachment to fibronectin-coated or subcutaneously implanted coverslips. Antimicrob Agents Chemother, 2000 Jun, 44(6), 1418 - 27 Identification and analysis of bacterial protein secretion inhibitors utilizing a SecA-LacZ reporter fusion system; Alksne LE et al.; Protein secretion is an essential process for bacterial growth, yet there are few if any antimicrobial agents which inhibit secretion . An in vivo, high-throughput screen to detect secretion inhibitors was developed based on the translational autoregulation of one of the central protein components, SecA . The assay makes use of a SecA-LacZ fusion reporter construct in Escherichia coli which is induced when secretion is perturbed . Several compounds, including two natural product extracts, which had the ability to induce the reporter fusion were identified and the MICs of these compounds for Staphylococcus aureus strain MN8 were found to be < or =128 microg/ml . Enzyme-linked immunosorbent assay, Western blotting, and immunoprecipitation techniques were used to analyze the affects of these compounds on protein secretion . Six representative compounds presented here appear to be bona fide secretion inhibitors but were found to have deleterious effects on membranes . It was concluded that, while the method described here for identifying inhibitors of secretion is valid, screens such as this, which are directed against the membrane-bound portion of a pathway, may preferentially identify compounds which affect membrane integrity. Antimicrob Agents Chemother, 2000 Jun, 44(6), 1413 - 7 Evernimicin (SCH27899) inhibits both translation and 50S ribosomal subunit formation in Staphylococcus aureus cells; Champney WS et al.; The effects of the everninomicin antibiotic evernimicin (SCH27899) on growing Staphylococcus aureus cells were investigated . Cellular growth rates and viable cell numbers decreased with increasing antibiotic concentrations . The rate of protein synthesis, measured as (35)S-amino acid incorporation, declined in parallel with the growth rate . Significantly, the formation of the 50S ribosomal subunit was inhibited in a dose-dependent fashion as well . 30S ribosomal subunit synthesis was not affected over the same concentration range . Evernimicin did not stimulate the breakdown of mature ribosomal subunits . Pulse-chase labeling experiments revealed a reduced rate of 50S subunit formation in drug-treated cells . Two erythromycin-resistant strains of S . aureus that carried the ermC gene were as sensitive as wild-type cells to antibiotic inhibition . In addition, two methicillin-resistant S . aureus organisms, one sensitive to erythromycin and one resistant to the macrolide, showed similar sensitivities to evernimicin . These results suggest a use for this novel antimicrobial agent against antibiotic-resistant bacterial infections. Microbes Infect . 2000 Apr;2(4):441. 'Infectious web'; The collagen-binding adhesin is a virulence factor in Staphylococcus aureus keratitis; Sid W . Richardson Ocular Microbiology Laboratory, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas, USAA collagen-binding strain of Staphylococcus aureus produced suppurative inflammation in a rabbit model of soft contact lens-associated bacterial keratitis more often than its collagen-binding-negative isogenic mutant . Reintroduction of the cna gene on a multicopy plasmid into the mutant helped it regain its corneal adherence and infectivity . The topical application of a collagen-binding peptide before bacterial challenge decreased S . aureus adherence to deepithelialized corneas . These data suggest that the collagen-binding adhesin is involved in the pathogenesis of S . aureus infection of the cornea. Infect Immun, 2000 Jun, 68(6), 3594 - 600 Monitoring bioluminescent Staphylococcus aureus infections in living mice using a novel luxABCDE construct; Francis KP et al.; Strains of Staphylococcus aureus were transformed with plasmid DNA containing a Photorhabdus luminescens lux operon (luxABCDE) that was genetically modified to be functional in both gram-positive and gram-negative bacteria . S . aureus cells containing this novel lux construct, downstream of an appropriate promoter sequence, are highly bioluminescent, allowing the detection of fewer than 100 CFU in vitro (direct detection of exponentially dividing cells in liquid culture) . Furthermore, these bacteria produce light stably at 37 degrees C and do not require exogenous aldehyde substrate, thus allowing S . aureus infections in living animals to be monitored by bioluminescence . Two strains of S . aureus 8325-4 that produce high levels of constitutive bioluminescence were injected into the thigh muscles of mice, and the animals were then either treated with the antibiotic amoxicillin or left untreated . Bioluminescence from bacteria present in the thighs of the mice was monitored in vivo over a period of 24 h . The effectiveness of the antibiotic in the treated animals could be measured by a decrease in the light signal . At 8 h, the infection in both groups of treated animals had begun to clear, as judged by a decrease in bioluminescence, and by 24 h no light signal could be detected . In contrast, both groups of untreated mice had strong bioluminescent signals at 24 h . Quantification of CFU from bacteria extracted from the thigh muscles of the mice correlated well with the bioluminescence data . This paper shows for the first time that bioluminescence offers a method for monitoring S . aureus infections in vivo that is sensitive and noninvasive and requires fewer animals than conventional methodologies. Infect Immun, 2000 Jun, 68(6), 3548 - 53 In vitro resistance of Staphylococcus aureus to thrombin-induced platelet microbicidal protein is associated with alterations in cytoplasmic membrane fluidity; Bayer AS et al.; Platelet microbicidal proteins (PMPs) are small, cationic peptides which possess potent microbicidal activities against common bloodstream pathogens, such as Staphylococcus aureus . We previously showed that S . aureus strains exhibiting resistance to thrombin-induced PMP (tPMP-1) in vitro have an enhanced capacity to cause human and experimental endocarditis (T . Wu, M . R . Yeaman, and A . S . Bayer, Antimicrob . Agents Chemother . 38:729-732, 1994; A . S . Bayer et al., Antimicrob . Agents Chemother . 42:3169-3172, 1998; V . K . Dhawan et al., Infect . Immun . 65:3293-3299, 1997) . However, the mechanisms mediating tPMP-1 resistance in S . aureus are not fully delineated . The S . aureus cell membrane appears to be a principal target for the action of tPMP-1 . To gain insight into the basis of tPMP-1 resistance, we compared several parameters of membrane structure and function in three tPMP-1-resistant (tPMP-1(r)) strains and their genetically related, tPMP-1-susceptible (tPMP-1(s)) counterpart strains . The tPMP-1(r) strains were derived by three distinct methods: transposon mutagenesis, serial passage in the presence of tPMP-1 in vitro, or carriage of a naturally occurring multiresistance plasmid (pSK1) . All tPMP-1(r) strains were found to possess elevated levels of longer-chain, unsaturated membrane lipids, in comparison to their tPMP-1(s) counterparts . This was reflected in corresponding differences in cell membrane fluidity in the strain pairs, with tPMP-1(r) strains exhibiting significantly higher degrees of fluidity as assessed by fluorescence polarization . These data provide further support for the concept that specific alterations in the cytoplasmic membrane of S . aureus strains are associated with tPMP-1 resistance in vitro. Infect Immun, 2000 Jun, 68(6), 3502 - 8 Role of interleukin-18 (IL-18) during lethal shock: decreased lipopolysaccharide sensitivity but normal superantigen reaction in IL-18-deficient mice; Hochholzer P et al.; Lethal shock can be associated with excessive secretion of cytokines such as tumor necrosis factor (TNF) and gamma interferon (IFN-gamma) . IFN-gamma mediates macrophage activation and appears to be controlled by interleukin (IL)-12 and IL-18 . To investigate the role of IL-18 in vivo, we generated IL-18-deficient mice by gene targeting . IL-18(-/-) mice showed decreased sensitivity towards lipopolysaccharide (LPS)-induced shock . LPS-induced IFN-gamma production was abrogated, yet induction of IL-12 and TNF was not affected . Both wild-type and IL-18-deficient mice succumbed to LPS-induced lethal shock after sensitization with D-galactosamine . However, in marked contrast to LPS, the bacterial superantigen Staphylococcus aureus enterotoxin B (SEB) induced comparable serum levels of IFN-gamma in IL-18(+/+) and IL-18(-/-) mice, accompanied by an upregulation of cell surface markers CD14, CD122 (IL-2Rbeta), and CD132 (IL-2Rgamma) on peritoneal macrophages . Moreover, SEB injection rendered IL-18-deficient mice sensitive for subsequent challenge with LPS . The degree of sensitization was comparable to that in wild-type controls with respect to lethality . However, LPS-induced TNF levels in serum were significantly reduced in SEB-sensitized IL-18-deficient mice . These results imply that IL-18 plays an important role in induction of IFN-gamma and lethality in response to LPS. Infect Immun, 2000 Jun, 68(6), 3261 - 8 Inhibition of staphylococcal enterotoxin B-induced lymphocyte proliferation and tumor necrosis factor alpha secretion by MAb5, an anti-toxic shock syndrome toxin 1 monoclonal antibody; Pang LT et al.; Toxic shock syndrome (TSS) is primarily caused by toxic shock syndrome toxin-1 (TSST-1) and staphylococcal enterotoxin B (SEB) . These toxins belong to a family of pyrogenic toxin superantigens (PTSAgs) produced by Staphylococcus aureus and exhibit several shared biological properties, including the induction of massive cytokine release and Vbeta-specific T-cell proliferation . The crystal structures of most PTSAgs are now published, and they demonstrate a striking similarity in conformational architecture even though their primary protein sequences are different . Despite these structural and immunobiological similarities, no cross-reactivity between TSST-1 and other PTSAgs has been demonstrated in serological or neutralization assays . Our laboratory has developed a neutralizing murine anti-TSST-1 monoclonal antibody (MAb5) which displayed cross-reactivity with SEB by enzyme-linked immunosorbent assay . The aim of the present study was to evaluate whether MAb5 can also cross-neutralize SEB-induced superantigenic activities in vitro . MAb5 was found to partially inhibit SEB-induced T-cell mitogenesis (63%) and tumor necrosis factor alpha (TNF-alpha) secretion (70%) in human peripheral blood mononuclear cells (PBMC) in a dose-dependent manner, while an isotypic anti-TSST-1 monoclonal antibody showed no effect . Epitope mapping revealed that MAb5 bound to TSST-1 residues 47 to 56 ((47)FPSPYYSPAF(56)) and to SEB residues 83 to 92 ((83)DVFGANYYYQ(92)), sequences that located in different regions of these toxins and are structurally dissimilar . SEB peptide (83)DVFGANYYYQ(92) was synthesized and found to also inhibit SEB-induced mitogenesis and TNF-alpha secretion in human PBMC . Our results demonstrate for the first time that MAb5 binds to different epitopes on TSST-1 and SEB that appear functionally important in inducing T-cell mitogenesis and TNF-alpha secretion in vitro. Biochem J, 2000 Jun 1, 348 Pt 2, 389 - 400 Purification from rat liver of a novel constitutively expressed member of the aldo-keto reductase 7 family that is widely distributed in extrahepatic tissues; Kelly VP et al.; Antiserum raised against human aflatoxin B(1) aldehyde reductase 1 (hAFAR1) has been used to identify a previously unrecognized rat aldo-keto reductase (AKR) . This novel enzyme is designated rat aflatoxin B(1) aldehyde reductase 2 (rAFAR2) and it characteristically migrates faster during SDS/PAGE than does the archetypal ethoxyquin-inducible rAFAR protein (now called rAFAR1) . Significantly, rAFAR2 is essentially unreactive with polyclonal antibodies raised against rAFAR1 . Besides its distinct electrophoretic and immunochemical properties, rAFAR2 appears to be regulated differently from rAFAR1 as it is expressed in most rat tissues and does not appear to be induced by ethoxyquin . Multiple forms of rAFAR2 have been identified . Anion-exchange chromatography on Q-Sepharose, followed by adsorption chromatography on columns of Matrex Orange A and Cibacron Blue, have been employed to purify rAFAR2 from rat liver cytosol . The Q-Sepharose chromatography step resulted in the resolution of rAFAR2 into three peaks of AKR activity, two of which were purified and shown to be capable of catalysing the reduction of 2-carboxybenzaldehyde, succinic semialdehyde, 4-nitrobenzaldehyde and 9,10-phenathrenequinone . The two most highly purified rAFAR2-containing preparations eluted from the Cibacron Blue column were 91 and 98% homogeneous . Analysis of these by SDS/PAGE indicated that the least anionic (peak CBA5) comprised a polypeptide of 37.0 kDa, whereas the most anionic (peak CBA6) contained two closely migrating polypeptides of 36.8 and 37.0 kDa; by contrast, in the present study, rAFAR1 was estimated by SDS/PAGE to be composed of 38.0 kDa subunits . Final purification of the 37 kDa polypeptide in CBA5 and CBA6 was accomplished by reversed-phase HPLC . Partial proteolysis of the two preparations of the 37 kDa polypeptide with Staphylococcus aureus V8 protease yielded fragments of identical size, suggesting that they represent the product of a single gene . Furthermore, the peptide maps from CBA5 and CBA6 differed substantially from that yielded by rAFAR1, indicating that they are genetically distinct from the inducible reductase . A peptide generated by CNBr digestion of the 37 kDa polypeptide from CBA6 was shown by Edman degradation to share 88% sequence identity with residues Tyr(168)-Leu(183) of rAFAR1 . This provides evidence that the rat protein identified by its cross-reactivity with anti-hAFAR1 serum is an additional member of the AKR7 family. Microbes Infect, 1999 Aug, 1(10), 795 - 805 Antimicrobial resistance in Staphylococcus aureus; Smith TL et al.; Recognized since 1883 as a common cause of infection, Staphylococcus aureus' preantimicrobial-era bacteremia mortality rate was 82% . The mortality of that era threatens to return as evidence of growing vancomycin resistance undermines the utility of vancomycin therapy . Successful treatment of S . aureus infections requires knowledge of its antimicrobial resistance capacity. Microbes Infect, 1999 Aug, 1(10), 745 - 51 Integrin-associated protein (IAP)-deficient mice are less susceptible to developing Staphylococcus aureus-induced arthritis; Verdrengh M et al.; The integrin-associated protein (IAP) has been shown to function in a signaling complex with beta3 integrins, influencing the migration of phagocytic cells into inflamed tissues . We have previously shown that gene-targeted mice deficient for IAP succumbed to peritonitis when inoculated with gram-negative bacteria . The aim of this study was to assess the role of IAP in our recently established model of haematogenously induced Staphylococcus aureus septicaemia and arthritis . In this model, neutrophils play a crucial role in the early phase of the infection . Mice lacking IAP and congenic controls were intravenously inoculated with S . aureus LS-1 . The IAP-/- mice were resistant to developing clinical signs of arthritis compared with their IAP-expressing littermates . The clinical findings were corroborated by histopathological evaluation indicating that the IAP-/- mice had less cartilage and bone destruction in the joints . We believe that a delayed migration of leukocytes into the joints of mice lacking IAP expression leads to decreased susceptibility to develop S . aureus-induced arthritis. Rev Med Suisse Romande, 2000 Mar, 120(3), 263 - 7 {Atopic dermatitis: the first allergic step in children}; Hofer MF; Atopic dermatitis is an early manifestation of atopy and is frequently present already during the first year of life . Atopic dermatitis is often associated with increased levels of serum IgE and with sensitization to food allergens . Some foods may be responsible for exacerbations of skin inflammation, but their pathogenic role need to be clinically assessed before an avoidance diet is prescribed . Staphylococcus aureus, which is known to colonize the skin of atopic dermatitis patients, may also exacerbate skin lesions and need to be treated with topical antibiotics . Children with atopic dermatitis are prone to allergies and are at risk to develop later respiratory allergic manifestations . In particular, if a sensitization to egg is present early in life, the risk for developing later an asthma due to house dust mites is increased . The care of children with atopic dermatitis should not be limited to the treatment of skin lesions, but should also involve preventive measures for respiratory allergies. J Org Chem, 2000 Jan 28, 65(2), 459 - 63 Chloptosin, an apoptosis-inducing dimeric cyclohexapeptide produced by Streptomyces; Umezawa K et al.; In the course of screening for apoptosis-inducing agents, chloptosin (1) was isolated from the culture broth of Streptomyces . The dumbbell-type structure of the dimeric cyclohexapeptide consisting of D-valine, (3S)- and (3R)-piperazic acids, O-methyl-L-serine, D-threonine, and (2S,3aR,8aR)-6-chloro-3a-hydroxy-2,3,3a, 8a-hexahydropyrrolo{2,3-b}indole-2-carboxylic acid was elucidated by spectroscopic and chemical degradation studies . The amino acid components in each cyclohexapeptide domain were presented in alternating R and S configurations . Chloptosin (1) was found to induce apoptotic activity in apoptosis-resistant human pancreatic adenocarcinoma cell line AsPC-1 and showed a strong antimicrobial activity against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus. Eur J Clin Invest, 2000 May, 30(5), 460 - 6 Effect of preoperative prophylaxis with filgrastim in cancer neck dissection; Wenisch C et al.; BACKGROUND: Cancer surgery is known to lead to a deterioration in host defence mechanisms and an increase in susceptibility to infection after operation . Filgrastim enhances important antimicrobial functions of neutrophils including chemotaxis, phagocytosis and oxidative killing mechanisms . METHODS: The effects of additional (all patients received perioperative 3 ' 25 mg kg-1 cefotiam and 1 ' 20 mg kg-1 metronidazole) preoperative prophylaxis with filgrastim (5 microg kg-1 12 h prior to surgery plus 5 microg kg-1 0 h prior to surgery) on neutrophil phagocytosis and reactive oxygen radical production and postoperative infections in 24 patients undergoing cancer neck dissection were studied . Phagocytic capacity was assessed by measuring the uptake of fluorescein isothiocyanate-labelled Escherichia coli and Staphylococcus aureus by flow cytometry . Reactive oxygen generation after phagocytosis was estimated by determining the amount of dihydrorhodamine 123 converted to rhodamine 123, intracellularly . RESULTS: In the filgrastim-treated patients a higher neutrophil phagocytic capacity was seen intraoperatively, and 1-5 days postoperative, but not prior to surgery . Reactive oxygen radical production was significantly higher in filgrastim-treated patients prior to surgery, intraoperative and postoperative (1-5 days) . 2/12 (17%) patients had postoperative infections in the filgrastim group and 9/12 (75%) patients had infections in the placebo group (P < 0.001) . In particular, wound infections were recorded more often in the placebo group (1/12 vs . 6/12; P = 0.004) . CONCLUSION: We conclude that filgrastim enhances perioperative neutrophil function and could be useful in the prophylaxis of postoperative wound infections in patients undergoing cancer neck dissection. J Bacteriol, 2000 Jun, 182(11), 3197 - 203 Identification, cloning, and initial characterization of rot, a locus encoding a regulator of virulence factor expression in Staphylococcus aureus; McNamara PJ et al.; A chromosomal insertion of transposon Tn917 partially restores the expression of protease and alpha-toxin activities to PM466, a genetically defined agr-null derivative of the wild-type Staphylococcus aureus strain RN6390 . In co-transduction experiments, transposon-encoded erythromycin resistance and a protease- and alpha-toxin-positive phenotype are transferred at high frequency from mutant strains to agr-null strains of S . aureus . Southern analysis of chromosomal DNA and sequence analysis of DNA flanking the Tn917 insertion site in mutant strains revealed that the transposon interrupted a 498-bp open reading frame (ORF) . Similarity searches using a conceptual translation of the ORF identified a region of homology to the known staphylococcal global regulators AgrA and SarA . To verify that the mutant allele conferred the observed phenotype, a wild-type allele of the mutant gene was introduced into the genome of a mutant strain by homologous recombination . The resulting isolates had a restored agr-null phenotype . Virulence factor gene expression in mutant, restored mutant, and wild-type strains was quantified by measuring alpha-toxin activity in culture supernatant fluids and by Northern analysis of the alpha-toxin transcript . We named this ORF rot (for repressor of toxins) (GenBank accession no . AF189239) because of the activity associated with rot::Tn917 mutant strains. Pharmacotherapy, 2000 May, 20(5), 589 - 92 Response of complicated methicillin-resistant Staphylococcus aureus endocarditis to the addition of trovafloxacin; Eckart RE et al.; The newer fluoroquinolones have many properties such as safety, bioavailability, and tissue penetration that make them attractive in the therapy of complicated infections . Unfortunately, the rapid development of resistance by Staphylococcus aureus to ciprofloxacin has dampened interest in these agents for serious staphylococcal infections . A patient with right-sided methicillin-resistant Staphylococcus aureus (MRSA) endocarditis with a complicated clinical course received trovafloxacin in addition to vancomycin and rifampin . He was initially treated with vancomycin, gentamicin, and rifampin for serious MRSA infection, but because of complications, including septic central nervous system emboli, persistent fever, and leukocytosis, gentamicin was stopped and trovafloxacin begun . After this addition the patient improved and completely recovered . In vitro and animal model data show that many newer fluoroquinolones have excellent activity against S . aureus, including MRSA, and are also less likely to induce resistance . Animal models of endocarditis support their efficacy in serious staphylococcal infections. J Nucl Med, 2000 May, 41(5), 912 - 8 Improved imaging of infections by avidin-induced clearance of 99mTc-biotin-PEG liposomes; Laverman P et al.; This article describes the preparation and optimization of biotin-polyethyleneglycol (PEG) liposomes and their application in experimental infection models to improve the scintigraphic imaging of infection and inflammation . METHODS: Biotin was coupled to PEG-distearoylphosphatidylethanolamine (DSPE) and subsequently incorporated in the PEG liposomes . Biotinylated liposomes were radiolabeled with 99mTc-hydrazinonicotinamide . In vitro binding studies were performed to find the optimal biotin concentration in the liposomes . In rats the biodistribution of the 99mTc-biotin-PEG liposomes was compared with the biodistribution of normal (nonbiotinylated) 99mTc-PEG liposomes . Furthermore, in vivo studies in rats were performed to study both the effect of the biotin content and the optimal avidin dose for efficient clearance of the liposomes . Liposomes containing 0.5 or 1.0 mol% biotin-PEG-DSPE were compared in rats with a Staphylococcus aureus infection in the left calf muscle . Avidin was injected 4 h after injection of the liposomes . RESULTS: Biotinylation of the liposomes did not affect their in vivo behavior . All biotin-PEG liposome formulations tested showed good in vitro avidin binding with 50% inhibitory concentrations ranging from 36 to 8 micromol/L . With avidin doses higher than 100 microg, both preparations rapidly cleared from the circulation . As a result, abscess-to-blood ratios increased 5-fold . To illustrate the potential of the avidin-induced clearance of radiolabeled PEG liposomes, we also studied the 99mTc-biotin-PEG liposomes in rabbits with a subcutaneous S . aureus abscess . The infection was visualized only after injection of 100 microg avidin . CONCLUSION: This study shows that biotin-coated 99mTc-PEG liposomes in combination with the injection of avidin can lead to improved imaging of infection or inflammation localized especially in regions with high blood-pool activity. J Microbiol Immunol Infect, 2000 Mar, 33(1), 57 - 9 Community-acquired methicillin-resistant Staphylococcus aureus endocarditis with septic embolism of popliteal artery: a case report; Lin JC et al.; A 20-year-old man presented with a 14-day course of fever . Physical examination showed petechiae of the conjunctivae, Janeway lesions on both hands, a grade III/VI systolic murmur over the apex, pulseless dorsal pedal artery and posterior tibial artery of the right leg, and a pale right foot . Femoral arteriogram of the right leg revealed total occlusion of the popliteal artery with collateral circulation of the posterior tibial artery . Transthoracic echocardiogram showed trace mitral regurgitation . Embolectomy of the right popliteal artery was done, and penicillin and gentamicin treatment was administered . However, postoperative fever developed intermittently . Transesophageal echocardiogram disclosed vegetation over the anterior leaflet of the mitral valve . Methicillin-resistant Staphylococcus aureus (MRSA) was isolated from all three cultures of blood drawn at admission and from the septic embolus during operation . He had neither evidence of underlying heart disease, nor history of intravenous drug abuse or hospitalization . Exploratory cardiotomy with removal of vegetation on the mitral valve was performed followed by a 4-week treatment with intravenous vancomycin . After discharge, he was well at 2-year follow-up. No Shinkei Geka, 2000 May, 28(5), 429 - 34 {Two cases of methicillin-resistant Staphylococcus aureus (MRSA) sepsis following craniotomy}; Kasuya H et al.; We report here two cases of MRSA sepsis following craniotomy . In case 1, a petroclival meningioma was subtotally removed and lumbar drainage was inserted postoperatively to prevent cerebrospinal fluid leakage . Ventriculo-peritoneal shunt was performed after meningitis was treated with vancomycin and panipenem/betamipron . Two weeks after the procedure, the patient revealed continuous spiking fevers related to MRSA sepsis, which did not improve with vancomycin and arbekacin administration . The focus of infection was found by scintigraphy and CT by 67Ga to be spondylo-diskitis at the level of L2-L3 . The lesion was removed and bone from the iliac crest grafted . In case 2, seven days after surgery for multiple meningioma, the patient exhibited spiking fevers and swelling in the left leg . The central venous catheter was removed from the left femoral vein and MRSA was found from blood culture . The patient was treated with arbekacin (200 mg/day) . Venous thrombosis diagnosed by CT was treated with heparin . Symptoms related to the infection and laboratory data did not improve because the concentration of arbekacin in the blood did not reach an effective level . The symptoms markedly improved when the dose of arbekacin was doubled (400 mg/day). Bioorg Khim, 2000 Jan, 26(1), 25 - 30 {Analytical biotechnology of recombinant peptides and proteins . I . Analysis of the purity, composition and structure of human, porcine and bovine insulins}; Sergeev NV et al.; A method for analysis of the type, purity, and possible structural modifications of insulins of bovine, porcine, and human origin was proposed . It is based on a combination of narrow-bore reversed-phase HPLC and mass spectrometry . The hydrolysis of insulins with highly specific Glu-protease V8 from Staphylococcus aureus followed by peptide mapping of the hydrolysis products and mass spectrometry of the isolated fragments helps rapidly and reliably localize and identify substitutions of amino acid residues in insulin structure by using insulin samples of less than 1 nmol. Proc Natl Acad Sci U S A, 2000 May 9, 97(10), 5510 - 5 Staphylococcus aureus sortase mutants defective in the display of surface proteins and in the pathogenesis of animal infections; Mazmanian SK et al.; Many gram-positive bacteria covalently tether their surface adhesins to the cell wall peptidoglycan . We find that surface proteins of Staphylococcus aureus are linked to the cell wall by sortase, an enzyme that cleaves polypeptides at a conserved LPXTG motif . S . aureus mutants lacking sortase fail to process and display surface proteins and are defective in the establishment of infections . Thus, the cell wall envelope of gram-positive bacteria represents a surface organelle responsible for interactions with the host environment during the pathogenesis of bacterial infections. Proc Natl Acad Sci U S A, 2000 May 9, 97(10), 5399 - 404 Crystal structure of a Staphylococcus aureus protein A domain complexed with the Fab fragment of a human IgM antibody: structural basis for recognition of B-cell receptors and superantigen activity; Graille M et al.; Staphylococcus aureus produces a virulence factor, protein A (SpA), that contains five homologous Ig-binding domains . The interactions of SpA with the Fab region of membrane-anchored Igs can stimulate a large fraction of B cells, contributing to lymphocyte clonal selection . To understand the molecular basis for this activity, we have solved the crystal structure of the complex between domain D of SpA and the Fab fragment of a human IgM antibody to 2.7-A resolution . In the complex, helices II and III of domain D interact with the variable region of the Fab heavy chain (V(H)) through framework residues, without the involvement of the hypervariable regions implicated in antigen recognition . The contact residues are highly conserved in human V(H)3 antibodies but not in other families . The contact residues from domain D also are conserved among all SpA Ig-binding domains, suggesting that each could bind in a similar manner . Features of this interaction parallel those reported for staphylococcal enterotoxins that are superantigens for many T cells . The structural homology between Ig V(H) regions and the T-cell receptor V(beta) regions facilitates their comparison, and both types of interactions involve lymphocyte receptor surface remote from the antigen binding site . However, T-cell superantigens reportedly interact through hydrogen bonds with T-cell receptor V(beta) backbone atoms in a primary sequence-independent manner, whereas SpA relies on a sequence-restricted conformational binding with residue side chains, suggesting that this common bacterial pathogen has adopted distinct molecular recognition strategies for affecting large sets of B and T lymphocytes. Eur J Nucl Med, 2000 Apr, 27(4), 392 - 9 Scintigraphic detection of acute experimental endocarditis with the technetium-99m labelled glycoprotein IIb/IIIa receptor antagonist DMP444; Oyen WJ et al.; Bacterial endocarditis is an important clinical problem that may result in persistent bacteraemia and irreversible cardiac damage . Since endocarditis is characterized by aggregation of activated platelets, fibrin and bacteria, we studied DMP444, a technetium-99m labelled high-affinity antagonist of the GP IIb/IIIa receptor that is expressed on activated platelets . In seven Beagle dogs (11-15 kg), the left ventricle was catheterized via the right carotid artery . One hour later, 5x10(7) colony forming units of Staphylococcus aureus were injected intracardially . Half an hour later, the catheter was removed . Two extra dogs underwent a complete sham procedure . One day after the intervention, five infected and the two non-infected dogs were injected with 37 MBq/kg 99mTc-DMP444 and two infected dogs with 37 MBq/kg 99mTc-IgG (used as a non-specific control agent) and imaged up to 4 h after injection . Samples were obtained for tissue counting, microbiology and histology . From 1 to 2 h post injection onward, there was clear focal accumulation of DMP444 in the aortic valve region when endocarditis was present, and this accumulation increased with time . The non-infected and the 99mTc-IgG injected dogs showed only persisting blood pool activity without any focal abnormality . At 4 h post injection, the in vivo valve-to-blood pool ratios were 1.87+/-0.18 in endocarditis, 1.01+/-0.05 in non-infected controls and 1.09+/-0.02 in 99mTc-IgG injected dogs (P<0.05) . It is concluded that targeting activated platelets with the 99mTc-labelled GP IIb/IIIa antagonist DMP444 allows a final diagnosis of experimental bacterial endocarditis within 4 h owing to high, specific and fast in vivo uptake. Rinsho Byori, 2000 Jan, Suppl 111, 69 - 74 {Detection methods for drug-resistant bacteria in routine examination--VISA}; Kinoshita S et al.; MRSA strains which clinically showed low-level resistance to vancomycin(VISA, hetero-VRSA) were isolated . Vancomycin-intermediate Staphylococcus aureus(VISA) was found to show a vancomycin minimum inhibitory concentration(MIC) of 8 to 16 micrograms/ml . The subpopulation of hetero vancomycin resistant MRSA(hetero-VRSA) was resistant to vancomycin MIC of 1 to 4 micrograms/ml . VISA and hetero-VRSA did not have vanA, vanB, vanC genes from vancomycin-resistant enterococci(VRE) . The disk diffusion test could not detect VISA and hetero-VRSA . The MIC tests can confirm VRSA well . To detect the hetero-VRSA, we need to use population analysis and growth on hetero-VRSA agar (MU3 agar plate). Rinsho Byori, 2000 Jan, Suppl 111, 40 - 7 {Detection methods for drug-resistant bacteria in routine examination--MRSA}; Higurashi Y et al.; Since methicillin-resistant Staphylococcus aureus(MRSA) is resistant not only to methicillin but also to multiple species of antibacterial drugs, there are cases where infections with MRSA are difficult to treat . For laboratory identification of MRSA, S . aureus is first identified, then an oxacillin (MPIPC) sensitivity test, PCR detection of the S . aureus mecA gene and an agglutination test using the latex sensitized with an anti-PBP-2' monoclonal antibody are usually utilized . However, the detection of MRSA does not necessarily indicate MRSA infection, and many cases only demonstrate MRSA colonization . Thus, a clinical investigation is indispensable for establishing the diagnosis of MRSA infection . In addition, recent reports also suggested a current trend toward decreasing sensitivity to Bactroban(MUP, mupirocin, an anti-intranasal MRSA antibacterial drug) . Taking these into consideration, surveillance of the trend in patient's sensitive to various species of antibacterial drugs including MUP must continue further. Org Lett, 2000 Apr 20, 2(8), 1081 - 4 Synthesis of an anti-methicillin-resistant Staphylococcus aureus (MRSA) carbapenem via stannatrane-mediated Stille coupling; Jensen MS et al.; {formula: see text} A short synthesis of carbapenem 1 is described . They key step involves the cross-coupling of an enol triflate with an amino-substituted sp3 carbon . This cross-couping, which allows the introduction of the complete side chain in one step, utilizes a stannatrane as the heteroalkyl transfer reagent. Microbiol Immunol, 2000, 44(2), 97 - 104 Staphylococcus aureus produces autolysin-susceptible cell walls during growth in a high-NaCl and low-Ca2+ concentration medium; Ochiai T; The growth of Staphylococcus aureus 209P becomes unusually sensitive to a high-NaCl concentration by decreasing the Ca2+ concentration in growth media, and cells either autolyze or transform into protoplast-like forms when grown standing in high-NaCl and low-Ca2+ concentration media below 37 C (Ochiai, T., Microbiol . Immunol . 43 (7): 705-709, 1999) . To assess the role of Ca2+ in the salt tolerance of this organism, cells grown in the presence of different concentrations of Ca2+ were treated with boiling SDS, and their susceptibilities to crude autolysin (3 M LiCl extract of S . aureus 209P cells) were evaluated by turbidimetric assay and zymographic analysis . Susceptibilities of SDS-treated cells (SDS-cells) to crude autolysin were significantly influenced by Ca2+ concentration in the culture, and SDS-cells prepared from cultures grown in high-NaCl and high-Ca2+ concentration media exhibited marked resistance to crude autolysin when the assay system contained a high concentration of NaCl . On the contrary, SDS-cells prepared from cultures grown in high-NaCl and low-Ca2+ concentration media were rather susceptible to crude autolysin under the same assay conditions . A zymographic analysis revealed that the constitution of cell-associated autolysins was not influenced by the concentration of exogenous Ca2+ . These results suggested that at least part of the mechanism of salt-induced autolysis in S . aureus 209P might be related to the synthesis of an autolysin susceptible cell wall. Med Dosw Mikrobiol, 1999, 51(3-4), 207 - 12 {Use of pulse-field electrophoresis for intraspecies differentiation of methycillin-resistant, coagulase-negative strains of Staphylococcus aureus (MRSA-CN)}; Mlynarczyk A et al.; Strains showing a negative reaction in tube test for coagulase accounted 10 to 20% of all Staphylococcus aureus isolated from patients of PSK No . 1 in Warsaw . They are MRSA as well as MSSA strains . 37 coagulase-negative isolates of MRSA were examined using the method depending on digestion of whole cell DNA with SmaI enzyme and electrophoretic separation of the obtained fragments in the pulsed field (PFGE) . It was shown that majority of the strains (26 from 37) had individual, unique patterns of bands . However, the two groups of strains were also observed showing a great similarity . The larger group contained 8 strains which were obtained from patients from different wards and the smaller group contained 3 strains obtained from patients from one hospital ward . The obtained results showed that among coagulase-negative MRSA strains are not only sporadic strains but also strains with probably epidemic properties, derived from different clones. FEMS Microbiol Lett, 2000 May 15, 186(2), 151 - 6 Intracellular Staphylococcus aureus induces apoptosis in mouse osteoblasts; Tucker KA et al.; Staphylococcus aureus invades osteoblasts and is the primary cause of osteomyelitis . This study examined the ability of S . aureus to induce apoptosis in a mouse osteoblast cell line . The presence of intracellular S . aureus was demonstrated by transmission electron microscopy . Light microscopy was utilized to examine morphological changes in the osteoblasts following killing of extracellular bacteria . Cell rounding was observed, and dark centers due to condensation of chromatin were noted in cells in infected osteoblast cultures . DNA was isolated from infected osteoblast cultures, and electrophoresis revealed the laddering effect characteristic of cells undergoing apoptosis . Additionally, an in situ cell death detection assay was utilized to label apoptosis-induced DNA strand breaks . Apoptotic nuclei were present, providing further evidence that S . aureus induces apoptosis in osteoblasts. Microbiol Immunol, 2000, 44(2), 79 - 88 Characterization and distribution of a new enterotoxin-related superantigen produced by Staphylococcus aureus; Abe J et al.; Staphylococcal enterotoxins (SEs) are a family of structurally related pyrogenic exotoxins consisting of the five prototypic SEs (types A to E) and three newly characterized SEs (types G to I) produced by Staphylococcus aureus (S . aureus) . They also work as superantigens and cause food poisoning and shock symptoms in humans . In this study, we cloned a new variant gene of the seg and characterized its superantigenic properties and distribution among the clinical isolates of S . aureus . The gene encodes a 233 amino acid protein which is highly homologous to SEG (97.7%) . The variant SEG (SEGv) expressed by the cloned gene exerted mitogenic activity on human peripheral blood mononuclear cells at the concentration of 100 pg/ml . T cells bearing Vbeta3, 12, 13.1, 13.2, 14 and 15 were preferentially expanded after stimulation with the recombinant protein . The mRNA of the variant seg gene was detected in the total RNA of the organisms bearing this gene . By PCR, 27 out of 48 clinical isolates of S . aureus (56%) possessed either the seg or variant seg gene . These findings suggest that SEG, or SEGv, is one of the most frequently produced superantigen exotoxins by S . aureus and may participate in the inflammatory process of the host by activating a distinct set of Vbeta families of T cells. Mol Cell Probes, 2000 Apr, 14(2), 71 - 8 Identification of Staphylococcus aureus enterotoxins A and B genes by PCR-ELISA; Gilligan K et al.; We developed PCR-enzyme linked immunosorbent (ELISA) assays to detect Staphylococcus aureus enterotoxins A and B genes . The assays use internal biotin-labelled oligonucleotides as capture probes for immobilizing and subsequently detecting target sequences on microtiter plates . The detection limits of the PCR-ELISAs were approximately 250 gene copies, versus 2500 gene copies by agarose gel analysis . The sensitivity of the assays, as determined from a reference panel of 46 coded samples that included DNA purified from 31 different bacterial species and strains, SEA and SEB plasmid controls, and no-template controls was 100% . No cross-reactivity was observed with DNA from non-staphylococcal species . Using 27 clinical isolates of S . aureus, the SEA PCR-ELISA identified the enterotoxin A (sea) gene in 26 samples, and the SEB PCR-ELISA identified the enterotoxin B (seb) gene in all 27 samples . Compared with conventional antigen capture ELISAs for SEA and SEB toxins, the PCR-ELISAs showed overall superior detection limits . The sensitivity and specificity levels of the SEA PCR-ELISA and the SEA toxin ELISA were comparable within their respective detection thresholds, but the sensitivity and specificity of the SEB PCR-ELISA was much greater than that of SEB toxin ELISA . Invest Ophthalmol Vis Sci, 2000 May, 41(6), 1432 - 7 Lysostaphin treatment of methicillin-resistant Staphylococcus aureus keratitis in the rabbit; Dajcs JJ et al.; PURPOSE: To determine the efficacy of lysostaphin treatment of methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MRSA) keratitis in a rabbit model . METHODS: The sensitivity to lysostaphin and vancomycin were compared for 34 MRSA and 12 methicillin-sensitive strains . Methicillin-resistant S . aureus strain 301 (MRSA 301) or a methicillin-sensitive strain of low virulence, ISP546, was intrastromally injected into rabbit corneas . Rabbit eyes were treated topically every 30 minutes from 4 to 9 or 10 to 15 hours postinfection with 0.28% lysostaphin or 5.0% vancomycin . Rabbits were killed and corneas were excised and cultured to determine the number of colony forming units (CFU) per cornea . RESULTS: Ninety percent minimal inhibitory concentrations were at least 19-fold lower for lysostaphin than for vancomycin . With early therapy (4 -9 hours postinfection) lysostaphin sterilized all MRSA 301-infected corneas, whereas untreated corneas contained 6.52 log CFU/cornea (P < or = 0.0001) . Corneas infected with MRSA 301 and treated similarly with vancomycin retained 2.3 +/-0.85 log CFU/cornea, and none were sterile . When therapy was begun later (10-15 hours postinfection) the residual bacteria in lysostaphin-treated eyes were significantly less numerous than in vancomycin-treated eyes (0.58 +/- 0.34 vs . 5.83 +/- 0.16 log CFU/cornea, respectively; P < or = 0.0001) . Three experiments were performed to demonstrate that lysostaphin penetrated the cornea to kill bacteria in vivo; lysostaphin-treated eyes were found to recover from infection, bacteria that did not cause epithelial defects (ISP546) were susceptible to lysostaphin, and inhibition of lysostaphin when harvesting corneas did not alter the observed therapeutic values of lysostaphin . CONCLUSIONS: Lysostaphin is very effective in treating keratitis mediated by methicillin-sensitive or methicillin-resistant S . aureus. J Clin Immunol, 2000 Jan, 20(1), 54 - 61 Relationship between interferon-gamma, interleukin-10, and interleukin-12 production in chronic hepatitis C and in vitro effects of interferon-alpha; Piazzolla G et al.; In the current study, increased interferon (IFN)-gamma, interleukin (IL)-10, and IL-12 p40 serum levels were observed in patients with chronic hepatitis C (CHC) compared to controls . Patients also displayed an increased spontaneous IFN-gamma release but a deficient peripheral blood mononuclear cells (PBMC) IFN-gamma production following stimulation with Staphylococcus aureus Cowan I strain (SAC) . No difference was found with reference to spontaneous or phytohaemagglutinin (PHA)-induced IL-10 release between patients and controls, whereas a higher IL-12 p70 and IL-12 p40 secretion triggered by SAC was observed in patients . Moreover, IL-12 p40/p70 ratio following SAC stimulation was higher in patients compared to controls and a negative correlation was found between this ratio and IFN-gamma amounts . Recombinant IL-12 (rIL-12) as well as neutralizing anti-IL-10 monoclonal antibodies (mAbs) were able to restore the compromised IFN-gamma production . Of note, anti-IL-10 supplementation induced a lower IL-12 p40/p70 ratio in HCV subjects as compared to controls . Finally, IFN-alpha upregulated in vitro IFN-gamma, IL-10, and IL-12 p70 release but not IL-12 p40 secretion, this giving rise to a normalization of IL-12 p40/p70 ratio . The data suggest the occurrence of an enhanced responsiveness to IL-10 modulating effects, likely mediated by an imbalance of IL-12 p40/p70 ratio, in chronic HCV infection . Cytokine balance restoration might thus contribute to achieve therapeutical results in chronic hepatitis C. J Med Microbiol, 2000 May, 49(5), 419 - 26 Characterisation of methicillin-resistant Staphylococcus aureus from Kuwait hospitals with high-level fusidic acid resistance; Udo EE et al.; Forty-seven fusidic acid- and methicillin-resistant Staphylococcus aureus isolates from clinical samples in four hospitals in Kuwait were studied for their relatedness by biotyping and pulsed-field gel electrophoresis (PFGE) and for the genetic location of their resistance determinants . Forty-four isolates were resistant to gentamicin, kanamycin and neomycin . Forty-one isolates were resistant to erythromycin and trimethoprim, 10 were resistant to chloramphenicol and four were resistant to ciprofloxacin . They contained two or three plasmids of c . 28, 2.8 and 1.8 kb . Genetic studies demonstrated that resistance to cadmium, propamidine isethionate and ethidium bromide were linked and were carried on the c . 28-kb plasmid . Chloramphenicol resistance was encoded by the 2.8-kb plasmid in resistant isolates . No resistance was associated with the 1.8-kb plasmid and this was considered to be a cryptic plasmid . Resistance to fusidic acid, methicillin, benzylpenicillin, gentamicin, kanamycin, neomycin, tetracycline, trimethoprim, erythromycin and ciprofloxacin were located on the chromosome . All the isolates produced urease, but varied in the production of haemolysins, pigments, lipase and lecithinase and were classified into nine biotypes . In contrast, PFGE divided the isolates into two major patterns with one PFGE type constituting the majority of isolates in all four hospitals . The presence of the dominant PFGE pattern in all four hospitals suggests that it is an epidemic MRSA clone with the capacity to spread . Infection control measures should be directed towards restricting the further spread of this clone. Indian J Pediatr, 1999 Jan-Feb, 66(1), 11 - 4 Microbial profile of neonatal infection in Coimbatore; Thomas M et al.; In this study, 187 consecutive neonates suspected of having septicaemia were investigated for isolation of micro organisms . Two samples of blood were collected for isolation of aerobes and anaerobes . Cultures were positive in 75 (40%) cases . Aerobic bacteria were the major etiological agent, accounting for 93% of positives including the 8% cases showing polymicrobial etiology . Anaerobic bacteria and Candida species were isolated in 6.6% and 8% of positive cases respectively . Bacteroides fragilis (amongst anaerobic) and Staphylococcus aureus (amongst aerobic) were the predominant organisms isolated . Clinical presentations were not specifically different to distinguish aerobic from anaerobic bacteria . In the present study, 6.6% of bacteremias were due to anaerobes, hence possibility of some of the bactermias being due to anaerobes should be kept in mind while treating cases of neonatal septicaemia . For a complete microbial profile both aerobic and anaerobic cultures should be done. Rev Med Interne, 2000 Apr, 21(4), 344 - 52 {Methicillin-resistant Staphylococcus aureus: factors responsible for its incidence}; Aubry-Damon H et al.; INTRODUCTION: How can we explain that the proportion of methicillin-resistant Staphylococcus aureus (MRSA) varies between the European countries, ranging from < 1% in Scandinavia to > 30% in Spain, France and Italy? This paper is aimed at attempting to determine factors at the origin of the spreading of endemic MRSA strains as of the early 1980s . Those strains are characterized by their ability to develop resistance to current antibiotics and make treatment of severe and deep infections more complex . CURRENT KNOWLEDGE AND KEY POINTS: Differences in the virulence of MRSA strains and that of susceptible strains appear unlikely . MRSA prevalence seems to be a growing problem, especially in Southern Europe where rates of resistance to other anti-staphylococcal antibiotics are high . General policies for antibiotic therapy as well as the implementation of strategies for prevention and control of MRSA might be responsible for such rates . Indeed, once MRSA is introduced into a facility without control program, this multiresistant bacteria rapidly spreads within the hospital and becomes endemic, expanding its reservoir . FUTURE PROSPECTS ET PROJECTS: Due to the introduction of new methods in microbiology and communication, infection control measures including procedures for isolation and identification of MRSA reservoirs are still feasible; however, their implementation requires human and material resources . Areas requiring improvement in the detection of MRSA outbreaks are identified in this paper, with particular emphasis on the need for national surveillance of MRSA prevalence and reappraisal of MRSA control strategies in French hospitals. Diagn Microbiol Infect Dis, 2000 May, 37(1), 57 - 62 Characterization of isolates associated with emerging resistance to quinupristin/dalfopristin (Synercid) during a worldwide clinical program; Dowzicky M et al.; Quinupristin/dalfopristin (Synercid) is an i.v . antibiotic active against serious Gram-positive infections . Its unique dual mode of action means that the potential for resistance development is expected to be low . To determine the incidence of in vitro emerging resistance in worldwide clinical studies, susceptibility to quinupristin/dalfopristin was measured for baseline pathogens and corresponding on- or post-study isolates from 880 evaluable patients . In comparative studies of community-acquired pneumonia, complicated skin and skin structure infections, and nosocomial pneumonia, the incidence of emerging resistance was low (1 of 453; 0.22%; 95% CI: 0 . 01-1.4%) . Resistance development occurred in only one pathogen (methicillin-resistant Staphylococcus aureus) . In noncomparative studies, six instances (1.8% of 338 evaluable cases; 95% CI: 0.7 to 4.0%) of emerging resistance (all vancomycin-resistant Enterococcus faecium) were confirmed, accompanied by therapeutic failure in four cases . Molecular typing did not confirm the identity of one pair of strains . Overall, the incidence of emerging resistance to quinupristin/dalfopristin was low. Protein Sci, 2000 Apr, 9(4), 734 - 41 An enigmatic peptide ligation reaction: protease-catalyzed oligomerization of a native protein segment in neat aqueous solution; Kumaran S et al.; We report an enigmatic peptide ligation reaction catalyzed by Glu-specific Staphylococcus aureus V8 protease that occurs in neat aqueous solution around neutral pH utilizing a totally unprotected peptide substrate containing free alpha-carboxyl and alpha-amino groups . V8 protease catalyzed a chain of ligation steps between pH 6 and 8 at 4 degrees C, producing a gamut of covalent oligomers (dimer through octamer or higher) of a native protein segment TAAAKFE (S39) derived from ribonuclease A (RNAse A) . Size-exclusion chromatography suggested the absence of strong interaction between the reacting peptides . The circular dichroism spectra of monomer through pentamer showed length-dependent enhancement of secondary structure in the oligomers, suggesting that protease-catalyzed ligation of a monomer to an oligomer resulted in a product that was more structured than its precursor . The relative conformational stability of the oligomers was reflected in their ability to resist proteolysis, indicating that the oligomerization reaction was facilitated as a consequence of the "conformational trapping" of the product . The ligation reaction proceeded in two phases-slow formation and accumulation of the dimer followed by a fast phase of oligomerization, implying that the conformational trap encountered in the oligomerization reaction was a two-step process . The Gly substitution at any position of the TAAAKFE sequence was deleterious, suggesting that the first step of the conformational trap, namely the dimerization reaction, that proceeded very slowly even with the parent peptide, was quite sensitive to amino acid sequence . In contrast, the oligomerization reaction of an Ala analog, AAAAKFE, occurred in much the same way as S39, albeit with faster rate, suggesting that Ala substitution stabilized the overall conformational trapping process . The results suggest the viability of the product-directed "conformational trap" as a mechanism to achieve peptide ligation of totally unprotected peptide fragments in neat aqueous solution . Further, the study projects the presence of considerable innate synthetic potential in V8 protease, baring rich possibilities of protein engineering of this enzyme to generate a "V8 peptide ligase." Pediatr Dermatol, 2000 Mar-Apr, 17(2), 111 - 4 Staphylococcal septicemia in children with atopic dermatitis; Hoeger PH et al.; Atopic dermatitis (AD) is frequently complicated by minor bacterial superinfections . Invasive infections such as osteomyelitis have rarely been reported . We describe two children with staphylococcal septicemia during an exacerbation of their AD . Cellulitis and underlying congenital heart disease, respectively, were considered predisposing factors for the development of bacteremia . Identical strains were isolated from the skin, and there was a significant increase in antibodies against Staphylococcus aureus capsular polysaccharide in one child . Our cases demonstrate the potential severity of bacterial skin infections in AD, especially when associated with an underlying condition that increases vulnerability to bacteremia . While their true incidence in children with AD is currently unknown, it is conceivable that systemic staphylococcal infections may be more common than previously thought . Staphylococcal bacteremia has to be considered in the differential diagnosis of fever in children with severe AD . Conversely, episodes of staphylococcal bacteremia should prompt a search for underlying predisposing factors. Br J Dermatol, 2000 Apr, 142(4), 680 - 7 Effects of two novel cationic staphylococcal proteins (NP-tase and p70)and enterotoxin B on IgE synthesis and interleukin-4 and interferon-gamma production in patients with atopic dermatitis; Jahreis A et al.; We have characterized the cell-mediated and humoral immune response of patients with atopic dermatitis (AD) and healthy controls in response to two novel staphylococcal antigens (NP-tase, p70) and the superantigen staphylococcal enterotoxin B (SEB) . The parameters studied were IgE, interleukin (IL)-4 and interferon (IFN)-gamma synthesis by peripheral blood mononuclear cells (PBMC) after stimulation with NP-tase, p70 and SEB in vitro . Both antigens, as well as SEB, induced IL-4 and IFN-gamma secretion in patients and controls . However, patients with AD showed a significantly diminished IFN-gamma production in response to NP-tase or SEB . Furthermore, we demonstrated a good correlation between antigen-stimulated IgE production and the IL-4/IFN-gamma ratio in vitro . A distinct subgroup of PBMC showed impaired IFN-gamma synthesis and enhanced IL-4 secretion after incubation with p70 or NP-tase . These data support evidence that a subgroup of patients with AD, synthesizing low levels of IFN-gamma after stimulation with staphylococcal antigens, may have impaired abilities to clear Staphylococcus aureus colonization . Persistent staphylococcal antigens could then be responsible for inflammatory and allergic skin reactions in patients with AD . We therefore conclude that, besides superantigens, staphylococcal antigens may also play a part in the pathogenesis of AD. J Dairy Sci, 2000 Apr, 83(4), 855 - 62 Antimicrobial susceptibility of Staphylococcus aureus isolated from bovine mastitis in Europe and the United States; De Oliveira AP et al.; Minimum inhibitory concentrations were determined for 811 strains of Staphylococcus aureus isolated from cases of bovine mastitis in 11 countries . The countries and number of isolates included Denmark (105), England (92), Finland (95), Germany (103), Iceland (22), Ireland (42), Norway (101), Sweden (123), Switzerland (69), United States (53), and Zimbabwe (6) . The antimicrobial agents tested were penicillin, ampicillin, oxacillin, cephalothin, ceftiofur, amoxicillin + clavulanate, penicillin + novobiocin, enrofloxacin, premafloxacin, erythromycin, clindamycin, lincomycin, pirlimycin, neomycin, lincomycin + neomycin, and sulfamethazine . The MIC90 for these antimicrobial agents for all strains were 0.5, 1.0, 1.0, 0.5, 1.0, < or =0.06, 0.125, 0.125, < or =0.0078, 0.5, 1.0, 16.0, 1.0, 2.0, 0.5, and 4.0 microg/ml, respectively . Overall, only small variations between countries were seen in the MIC90 for the majority of compounds tested . Of the strains tested, 35.6% were positive for beta-lactamase production on initial testing, with an additional 21.3% positive after induction by penicillin . In conclusion, the overall level of resistance was generally low for all antimicrobial agents tested regardless of country . Given the differences in antimicrobial use in various countries, the widespread adoption of mastitis control programs to prevent infections limits the exposure of S . aureus infected animals to antimicrobial drugs. J Orthop Trauma, 2000 Mar-Apr, 14(3), 206 - 11 Evaluation of standard surgical preparation performed on superficial dermal abrasions; Jeray KJ et al.; OBJECTIVES: To determine the difference, if any, between the reduction of bacteria on contaminated normal skin and contaminated superficially abraded skin following standard surgical preparations at clinically relevant time points after injury . DESIGN: Prospective animal study . SETTING: Laboratory . SUBJECTS: Thirty-two New Zealand white rabbits . INTERVENTION: Two sites, two by two centimeters, one abraded and one nonabraded (control), were studied on each rabbit . Both were inoculated with encapsulated Staphylococcus aureus strain Wood 46 . Four six-millimeter punch biopsies were obtained after inoculation, immediately before surgical scrub, and five minutes and then two hours after completion of the surgical scrub . The rabbits were divided into four cohort groups with surgical scrubs performed at six, twelve, twenty-four, and forty-eight hours after inoculation . Bacterial counts were determined . MAIN OUTCOME MEASUREMENTS: Numbers of bacteria on surgical sites . RESULTS: Before surgical preparation, the amount of bacteria on the normal skin (control sites) dropped significantly (p<0.02) except in the six-hour group (p<0.20) . At the abraded skin sites, the bacteria flourished . The surgical scrub dropped bacterial counts at both the abraded and nonabraded skin sites significantly (p<0.05) except for the abraded site in the twenty-four-hour group (p<0.08) . However in the twelve-, twenty-four-, and forty-eight-hour groups, the bacterial counts (colony-forming units) were still markedly elevated (>1x10(5) at abraded sites) when compared with the nonabraded skin sites (p<0.008) at the respective time intervals . Only at the six-hour interval were the bacterial counts reduced similarly at both the abraded and nonabraded skin sites . CONCLUSIONS: In a rabbit model the standard surgical preparation using povidone-iodine at six hours after inoculation is effective in reducing the bacterial count on abraded skin to that of surgically prepared nonabraded skin . Beyond that time, the standard surgical preparation is ineffective in reducing counts to those of nonabraded skin at similar time intervals. J Clin Microbiol, 2000 May, 38(5), 1832 - 8 Transfer of erythromycin resistance from poultry to human clinical strains of Staphylococcus aureus; Khan SA et al.; The transfer of ermA and ermC genes, the two most common resistance determinants of erythromycin resistance, was studied with Luria-Bertani broth in the absence of additional Ca(2+) or Mg(2+) ions . Fifteen human and five poultry isolates of Staphylococcus aureus, which were resistant to erythromycin but carried different genetic markers for erythromycin resistance, were used for conjugation . Since both the donors (Amp(s)-Tet(r)) and recipients (Amp(r)-Tet(s)) were resistant to erythromycin, the transconjugants were initially picked up as ampicillin- and tetracycline-resistant colonies . The resistance transfer mechanisms of the chromosomally located erythromycin rRNA methylase gene ermA and the plasmid-borne ermC gene were monitored by a multiplex PCR and gene-specific internal probing assay . Four groups of transconjugants, based upon the transfer of the ermA and/or ermC gene, were distinguished from each other by the use of this method . Selective antibiotic screening revealed only one type of transconjugant that was resistant to ampicillin and tetracycline . A high frequency of transfer (4.5 x 10(-3)) was observed in all of the 23 transconjugants obtained, and the direction of tetracycline and erythromycin resistance marker transfer was determined to be from poultry to clinical isolates . The transfers of the ermA and ermC genes were via transposition and transformation, respectively. J Clin Microbiol, 2000 May, 38(5), 1797 - 803 Oxygen and carbon dioxide regulation of toxic shock syndrome toxin 1 production by Staphylococcus aureus MN8; Yarwood JM et al.; The production of toxic shock syndrome toxin 1 (TSST-1) by Staphylococcus aureus MN8 exposed to a range of oxygen concentrations (0 to 21% {vol/vol}) was examined in batch and thin-film cultures . The response of S . aureus to this range of oxygen concentrations was studied in the absence and in the presence of 7% (vol/vol) carbon dioxide . In the absence of carbon dioxide, TSST-1 production in batch cultures increased from negligible levels in the presence of oxygen concentrations of 1% or less to 500 ng/ml in the presence of 2% oxygen and then decreased to 70 ng/ml or less in the presence of oxygen concentrations of 6% and higher . In the presence of carbon dioxide, however, toxin production increased from negligible levels in the presence of 1% oxygen to 1,900 ng/ml in the presence of 21% oxygen . In thin-film cultures, TSST-1 production increased from nearly undetectable levels under anaerobic conditions to 1 and 10 microg/ml under 21% oxygen in the absence and presence of carbon dioxide, respectively . This study demonstrates the controlling effects of both oxygen and carbon dioxide on TSST-1 production. J Chemother, 2000 Apr, 12(2), 138 - 44 Analysis of a mini-outbreak of methicillin-resistant Staphylococcus aureus in a surgical ward by using arbitrarily primed-polymerase chain reaction; Cetinkaya Y et al.; In November 1995, an increase was noticed in the number of methicillin-resistant Staphylococcus aureus (MRSA) isolates from a surgical ward at Hacettepe University Hospital . All MRSA isolates obtained from clinical specimens in this ward (14 MRSA isolates from wound cultures of 10 patients) were collected prospectively for 10 weeks . Surveillance cultures were taken from ward personnel (nose cultures from 4 physicians, 7 nurses, 1 secretary, 1 waiter), 2 surgical dressing containers and 1 nebulizer . MRSA was isolated from one of the surgical dressing containers, the nebulizer and nose cultures of 3 physicians, 3 nurses and the ward secretary . Arbitrarily primed polymerase chain reaction (AP-PCR) analysis showed that most MRSA isolates belonged to 2 major clones (pattern A, pattern B) . Pattern A was the most frequent one and was present in 4 clinical isolates, surgical dressing container-1 . Pattern B was identified in 3 clinical isolates and nose culture of physician-3 . AP-PCR analysis revealed that this mini-MRSA outbreak was caused by contamination of surgical dressing container with MRSA and nasal MRSA carriage in ward staff . AP-PCR seems to be a valuable typing method for analysis of nosocomial MRSA outbreaks because of its simplicity and rapidity. Microbiol Immunol, 2000, 44(3), 189 - 91 An exfoliative toxin A-converting phage isolated from Staphylococcus aureus strain ZM; Yoshizawa Y et al.; Exfoliative toxin A (ETA) causes staphylococcal scalded-skin syndrome in children . The gene for ETA was believed to be coded by the chromosomal DNA . We isolated temperate phages from an ETA-producing strain, ZM, using a restriction minus strain, 1039, as an indicator . One of the prophages, designated phi-ZM-1 mediated lysogenic conversion of ETA . The polymerase chain reaction assay of the eta gene revealed that phage phi-ZM-1 carries the structural gene for ETA. Rev Neurol, 2000 Feb 16-29, 30(4), 326 - 8 {Subdural empyema due to Mycoplasma hominis following epidural anesthesia}; Escamilla F et al.; INTRODUCTION: In the literature there are sporadic reports of spinal epidural abscesses after epidural anaesthesia (Staphylococcus aureus in 82%), whilst subdural empyemas are more often related to ear and sinus conditions . CLINICAL CASE: A 32 year old woman with a clinical history of migraine and symmetrical frontal atrophy on a previous cerebral CT scan, after Caesarean section under epidural anaesthesia, presented with orthostatic headache two days later . On the fourth day it had become constant and she had a high temperature which was considered to be caused by infection of the surgical wound . Neurological examination was found to be normal, the CT scan was inconclusive and the CSF showed a lymphocytic pleocytosis without consumption of glucose . In view of her worsening clinical condition on the ninth day, in the absence of a cutaneous focus and on suspicion of a para-meningeal infective focus, lumbar MR was done and found to be normal, and cerebral MR which showed images compatible with a right fronto-parietal subdural empyema . After a parietal craniotomy and culture of the surgical specimen, colonies of Mycoplasma hominis were grown, similar to those grown from the exudates of the abdomical surgical wound . Treatment was started with ciprofloxacine . CONCLUSION: We consider that following epidural anaesthesia the patient developed hypotension of the CSF with a secondary subdural hematoma or hygroma and this became infected by hematogenous spread of Mycoplasma hominis. Braz J Infect Dis, 2000 Feb, 4(1), 36 - 42 Evaluation in an animal model and in vitro of the combination clavulanic acid and cephalosporins against beta-lactamase producing and nonproducing Staphylococcus aureus strains; de Sa Del Fiol F et al.; Beta-lactamase enzymes are the most common cause of bacterial resistance to Beta-lactam antibiotics . They hydrolyze the amide bound in the Beta-lactam ring and produce acidic derivatives that have no antibacterial properties . The aim of this study was to evaluate a combination of clavulanic acid with cephalosporins against Beta-lactamase-producing and nonproducing strains of Staphylococcus aureus using in vitro tests and a rat animal model . In vitro tests (MIC) of the drug combination were done using standard methods . In an animal model, rats were submitted to surgical implantation of polyurethane sponges in their backs to induce granulomatous tissue . After seven days, the animals received cephalexin, cephalexin with clavulanic acid, ceftriaxone, ceftriaxone with clavulanic acid or clavulanic acid alone . One hour after the drug administration, granulomatous tissue was removed and placed in Petri dishes previously inoculated with 10(8) cfu of producing or non-producing Beta-lactamase Staphylococcus aureus . After 24h at 37 degrees C, the inhibition zones formed by granulomatous tissue was measured and scored for statistical analysis . Both tests (ex vivo inverted question markanimal model inverted question mark and in vitro) showed that the cephalexin was more active than ceftriaxone against non-producing Beta-lactamase S.aureus (p<0.01) . Against Beta-lactamase producing S.aureus, ceftriaxone was more active than cephalexin, which was inactive . Combinations of clavulanic acid with cephalexin or ceftriaxone had similar antimicrobial activity against non-producing Beta-lactamase S.aureus compared to the cephalosporins used alone . When tested using Beta-lactamase producing strains, the combination of clavulanic acid with cephalosporins showed synergism . We conclude that the combination of cephalosporins with clavulanic acid could be useful in staphylococcal infections caused by Beta-lactamase producing strains. J Biol Chem, 2000 May 5, 275(18), 13863 - 71 The fibronectin-binding MSCRAMM FnbpA of Staphylococcus aureus is a bifunctional protein that also binds to fibrinogen; Wann ER et al.; Staphylococcus aureus is an important pathogen capable of causing a wide spectrum of diseases in humans and animals . This bacterium expresses a variety of virulence factors that participate in the process of infection . These include MSCRAMMs (microbial surface components recognizing adhesive matrix molecules) that mediate the adherence of the bacteria to host extracellular matrix components, such as collagen, fibronectin (Fn), and fibrinogen (Fg) . Two Fn-binding MSCRAMMs, FnbpA and FnbpB, have been previously identified . The Fn binding activity has been localized to the approximately 40-amino acid residue D repeats in the C-terminal part of these proteins . However, no biological activity has yet been attributed to the N-terminal A regions of these proteins . These regions exhibit substantial amino acid sequence identity to the A regions of other staphylococcal MSCRAMMs, including ClfA, ClfB, and SdrG (Fbe), all of which bind Fg . This raises the question of whether the Fn-binding MSCRAMMs can also bind specifically to Fg . In this report, we show that a recombinant form of the A region of FnbpA does specifically recognize Fg . We localize the binding site in Fg for recombinant FnbpA to the gamma-chain, in particular to the C-terminal residues of this polypeptide, the site also recognized by ClfA . In addition, we demonstrate that recombinant FnbpA can compete with ClfA for binding to both immobilized and soluble Fg . By the use of surface plasmon resonance spectroscopy and fluorescence polarization, we determine the dissociation equilibrium constant for the interaction of recombinant FnbpA with intact immobilized Fg and with a synthetic C-terminal gamma-chain peptide, respectively . Finally, by overexpressing FnbpA in a mutant strain of S . aureus that lacks the expression of both ClfA and ClfB, we show that native FnbpA can mediate the interaction of S . aureus with soluble Fg. World J Surg, 2000 May, 24(5), 528 - 31; discussion 532 Ultrastructural alterations of polytetrafluoroethylene prostheses implanted in abdominal wall provoked by infection: clinical and experimental study; Bellon JM et al.; Infection of an expanded polytetrafluoroethylene (ePTFE) prosthesis after implant is a major drawback of its use in current clinical practice . The aim of the present study was to compare the behavior of such prostheses implanted into New Zealand rabbits with that of prostheses infected after clinical implant . Experimental implants of ePTFE Soft Tissue Patch were performed to repair defects (7 x 5 cm) created in the abdominal wall of 10 rabbits . Prior to implant the prostheses were contaminated with Staphylococcus aureus . Five animals implanted with noncontaminated ePTFE prostheses served as controls . All the animals were sacrificed at 30 days after implant . For the clinical study, specimens were taken from three ePTFE implants that had been found to be infected after intervention . The clinical and experimental implant specimens were processed for light microscopy and scanning electron microscopy . Macroscopic and microscopic examination of experimental and clinical implants revealed alterations to the ePTFE structure, such as areas of fragmentation, fracture lines, and detachment of fine layers of ePTFE that harbored numerous Staphylococcus colonies . Neoformed tissue around contaminated implants was arranged more loosely, and on occasion large spaces between fibers gave rise to an "unknitted" appearance with respect to the control implants . It may be concluded that microporous ePTFE prostheses show similar behavior following experimental or clinical implant in the presence of infection . Irreversible changes to the structure of the prosthesis are produced owing to colonization of the biomaterial by microorganisms, which in most cases necessitates total replacement of the prosthesis. J Egypt Soc Parasitol, 2000 Apr, 30(1), 277 - 86 A study of the predisposition of Schistosomiasis mansoni to pyogenic liver abscess in experimentally infected mice; Mahmoud MS et al.; In murine experimental model, the present work was carried out to study the possible predisposition of S . mansoni infection to pyogenic liver abscess, through a concurrent infection with Staphylococcus aureus . Pyogenic liver abscesses containing the bacterial organism were found in 85% of mice infected with concurrent early S . mansoni infection and S . aureus, in 35% of mice with late S . mansoni infection and the bacterial infection, and in 60% of mice with concurrent immunosuppressed late St . mansoni infection and staphylococcal infection . The early S . mansoni infection, immunosuppressed late schistosomiasis mansoni, and to a less extant late schistosomiasis mansoni, with concurrent bacterial infections can predispose to pyogenic liver abscess which has implicative effect on clinical cases. Int J Immunopharmacol, 2000 Jul, 22(7), 557 - 66 Cefodizime enhances phagocytosis and intracellular killing of Staphylococcus aureus but does not influence polymorphonuclear leukocytes response to fMLP stimulation; Bialasiewicz P et al.; The influence of Cefodizime (CDZ) on in vitro activity of polymorphonuclear leukocytes (PMNL) from healthy subjects was assessed . Preincubation with CDZ enhanced phagocytosis and intracellular killing of Staphylococcus aureus by PMNL . Contrary to numerous clinical reports, no significant effect of CDZ preincubation on PMNL response to n-formyl-methionyl-leucyl-phenylalanine was found with respect to intracellular calcium changes, degranulation, hydrogen peroxide production, and chemiluminescence . These results suggest that augmented microbicidal activity of PMNL is not related to the enhanced production of reactive oxygen species in healthy subjects. Int J Biochem Cell Biol, 2000 Jun, 32(6), 665 - 75 Natural allelic variation of duck erythrocyte histone H1b; Palyga J et al.; In our previous work (J . Palyga, Genetic polymorphisms of histone H1 . b in duck erythrocytes . Hereditas 114, 85-89, 1991) we reported a genetic polymorphism of duck erythrocyte histone H1.b . Here, we screened H1 preparations in a two-dimensional polyacrylamide gel to refine the distribution of allelic forms of H1.b in fifteen duck populations . We have revealed that the frequency of H1.b allelic variants was significantly different among many conservative and breeding duck groups . While b(1) and b(3) were common in all populations screened, the allele b(2), with a slightly lower apparent molecular weight, was confined mainly to brown-feathered ducks (Khaki Campbell and Orpington) and descendent lines.The C- and N-terminal peptides released upon cleavage with N-bromosuccinimide and Staphylococcus aureus protease V8 from duck allelic histones H1 . b2 and H1.b3, respectively, migrated differently in the gel, probably as a result of potential amino acid variation in a C-terminal domain. Int J Biochem Cell Biol, 2000 Jun, 32(6), 597 - 608 Ovine placental lactogen and ovine prolactin: partial proteolysis and conformational stability; Fernandez ML et al.; The high-resolution structure of ovine placental lactogen (oPL) and ovine prolactin (oPRL), not yet established in detail, was probed by limited proteolysis with the Glu-specific protease from Staphylococcus aureus V8 . While in hGH there were no cleavage sites inside of any of the four alpha-helices, the analysis of the fragments obtained after partial proteolysis of oPL showed a site of cleavage at the putative third helix, suggesting that this helix is partially unwound at this point . The partial proteolysis of the rest of the molecule was compatible with a similar folding pattern for oPL, hGH and pGH, on the basis of the crystal structure of these last hormones . In the case of oPRL, proteolytic cleavage occurred at Glu residues which would be located at the end of the first helix and the beginning of the second in the hGH folding model, suggesting that these helices are shorter in oPRL than in hGH . In order to gain further insight on the folding of these molecules, circular dichroism and intrinsic fluorescence measurements were used to examine the effect of denaturing conditions on oPL and oPRL . After exposure to 6 M guanidine the unfolding of both proteins was completely reversed upon elimination of the denaturing agent . In contrast, exposure to pH 3.0 caused an irreversible decrease in the alpha-helical content in both hormones, most striking for oPL, indicating that this hormone is less stable than oPRL or hGH. Intensive Care Med, 2000 Feb, 26(2), 218 - 20 Transient EDTA-dependent pseudothrombocytopenia in a patient with sepsis; Mori M et al.; Ethylenediaminetetraacetic acid-dependent pseudothrombocytopenia (EDTA-PTCP) is the phenomenon of a spurious low platelet count due to antiplatelet antibodies that cause platelet clumping in blood anticoagulated with EDTA . We describe a case of EDTA-PTCP that appeared transiently with the development of sepsis . A 50-year-old man underwent Bentall's aortic root replacement for acute aortic dissection with aortic insufficiency . Postoperatively the patient suffered paralytic ileus followed by methicillin-resistant Staphylococcus aureus enteritis and septicemia with endotoxemia . EDTA-PTCP appeared with the development of sepsis, and disappeared with its resolution . To avoid incorrect diagnoses and inappropriate treatment, EDTA-PTCP should always be considered as a possible cause of reported low platelet counts, even in patients with sepsis. Microbiology, 2000 Apr, 146 ( Pt 4), 981 - 7 Identification of Staphylococcus aureus genes expressed during growth in milk: a useful model for selection of genes important in bovine mastitis? Lammers A, Kruijt E, van de Kuijt C, Nuijten PJ, Smith HE. Staphylococcus aureus is a major cause of bovine mastitis . Since gene expression of many bacteria is known to be regulated by the environment, milk may play an important role in the regulation of the early steps in the pathogenesis of bovine mastitis by S . aureus . To get insight into the response of S . aureus to the milk environment, a Tn917-lacZ mutant library was generated and screened for genes specifically expressed during growth in milk . Twenty-eight mutants were identified and analysed . Four groups of genes were found, involved in cell-wall synthesis, nucleotide synthesis, transcriptional regulation and carbohydrate metabolism . A fifth group contained genes with hypothetical or unknown functions . Many of the genes identified belonged to biosynthetic pathways of S . aureus and other bacterial species which have also been shown to play a role in vivo as determined in murine infection models . Therefore, growth on milk may be an attractive model for the identification of genes preferentially expressed during bovine mastitis. Infect Control Hosp Epidemiol, 2000 Apr, 21(4), 270 - 1 Evidence of delays in transferring patients with methicillin-resistant Staphylococcus aureus or vancomycin-resistant Enterococcus to long-term-care facilities; Bryce EA et al.; This retrospective case-control study examined whether there was a difference in length of time awaiting long-term-care placement for patients identified as having methicillin-resistant Staphylococcus aureus or vancomycin-resistant Enterococcus compared to controls . Thirty-nine patients with methicillin-resistant Staphylococcus aureus or vancomycin-resistant Enterococcus waited for placement an average of 61 days longer than controls (P<.0002) . The average number of requests for placement was 2.5 compared to 1.7 for controls (P=.015). Ann Fr Anesth Reanim, 2000 Mar, 19(3), 177 - 87 {Glycopeptides}; Leone M et al.; OBJECTIVES: To review pharmacology, pharmacokinetic and therapeutic use of glycopeptides in intensive care units . DATA SOURCES: Extraction from Medline database of French and English articles on glycopeptides and search along with major review articles . DATA SELECTION: The collected articles were reviewed and selected according to their quality and originality . The more recent data were selected . DATA SYNTHESIS: Glycopeptides are bactericidal antibiotics which are only active against Gram positive species acting by inhibiting peptidoglycan synthesis . They had been in clinical use for almost 30 years without high-level resistance underlining . For ten years, there have been disturbing reports of first, resistance to vancomycin in enterococcal species and more recently in strains of Staphylococcus aureus by complex and large mechanisms of action . This new resistances may lead to a therapeutic impasse and a fatal issue for infected patients . The only response to this situation is the respect of prescription rules and the careful use of antibiotics . CONCLUSION: Considering their spectrum, glycopeptides are an antibiotic family which importance is fundamental to treat infected patients of intensive care units . Staff members of intensive care units are responsible for their good use. Ann Fr Anesth Reanim, 2000 Mar, 19(3), 151 - 5 {Colonization by methicillin-resistant Staphylococcus aureus is a predictive factor for the resistance phenotype of an infectious strain of S . aureus}; Truffault A et al.; OBJECTIVE: To assess the predictive value of a previous colonization with methicillin-resistant Staphylococcus aureus for the resistance pattern of a bacteriological specimen significantly positive to S . aureus . STUDY DESIGN: Retrospective study of patients' files . PATIENTS: Patients admitted for at least 48 hours in a surgical intensive care unit from April 1, 1996 to December 31, 1997 . METHODS: Collection of patients' characteristics and chronology of positive microbiological specimens with methicillin-susceptible (MSSA) or -resistant (MRSA) S . aureus from medical and laboratory records . During the study period, screening for nasal or perineal colonization with MRSA was systematically performed on admission and weekly thereafter . RESULTS: The files of 540 patients were reviewed . MSSA and MRSA infections occurred in 7% (39/540) and 4% (20/540) of the patients respectively . By opposition with MSSA infections, MRSA infections occurred more frequently in patients previously colonized with MRSA (13 infections in 63 colonized patients {21%} versus 7 infections in 477 non-colonized patients {2%}, odds ratio = 18, confidence interval: 6-51, P < 0.0001) . The median delay between colonization and infection was 5 days . The positive and negative predictive values for previous colonization with MRSA to predict infection with MRSA in presence of a bacteriological specimen significantly positive with S . aureus were 81 and 84%, respectively . CONCLUSION: The probabilistic use of a glycopeptide in presence of a bacteriological specimen significantly positive with S . aureus should be limited to patients already colonized with MRSA, in order to decrease the abusive administration of these antibiotics. West J Med, 2000 Apr, 172(4), 235 - 9 Outbreak of boils in an Alaskan village: a case-control study; Landen MG et al.; OBJECTIVE: To determine whether taking steam baths was associated with furunculosis and to evaluate possible risk factors for the occurrence of boils during a large outbreak in Alaska . DESIGN: A cohort study of village residents, a case-control study, and assessment of environmental cultures taken from steam baths . SETTING: Village in southwestern Alaska . PARTICIPANTS: 1 adult member from 77 of the 92 households in the village was interviewed; 115 residents with at least one boil occurring between January 1 and December 12, 1996 were considered to be cases; 209 residents without a boil acted as the control group . All 459 village residents were included in the cohort study . MAIN OUTCOME MEASURE: Rate of infection among all residents and residents who regularly took steam baths, risk factors for infection, and relative risk of infection . RESULTS: 115 people (25%) had had at least one boil . Men were more likely to have had a boil than women (relative risk 1.5; 95% confidence interval 1.1 to 2.2) . The highest rate of infection was among people ages 25-34 years (32/76; 42%) . No children younger than 2 years had had boils . Boils were associated with using a steam bath (odds ratio 8.1; 3.3 to 20.1) . Among those who used a steam bath, the likelihood of developing boils was reduced by routinely sitting on a towel while bathing, which women were more likely to do, and bathing with fewer than 8 people . Of the 93 samples taken from steam baths, one Staphylococcus aureus isolate was obtained from a bench in an outer dressing room . CONCLUSION: Using a steam bath was associated with developing boils in this outbreak in a village in Alaska . People should be advised to sit on towels while using steam baths. Scand J Rheumatol, 2000, 29(2), 133 - 5 Acute non-purulent inflammatory arthropathy associated with Staphylococcus aureus abscess; Al-Allaf AW et al.; Case 1 . A 20-year-old woman presented 4 weeks post-partum with widespread symmetrical inflammatory polyarthropathy with marked synovitis . Investigations revealed grossly raised CRP with negative immunology screen . A few days before presentation she saw her general practitioner with left-sided mastitis, which then developed into a Staphylococcus breast abscess . Surgical drainage of this led to almost immediate resolution of the joint complaints and return of CRP to normal . Case 2 . A 27-year-old man developed widespread symmetrical inflammatory arthropathy . A few days prior to this he had developed folliculitis with a furuncle on his neck . Swab grew Staphylococcus aureus . His arthritis settled immediately following spontaneous drainage of his abscess and a full course of antibiotic . The pathogenic mechanism is unclear but could be toxin-mediated. Eur J Nucl Med, 2000 Mar, 27(3), 292 - 301 Technetium-99m labelled antimicrobial peptides discriminate between bacterial infections and sterile inflammations; Welling MM et al.; The aim of this study was to select technetium-99m labelled peptides that can discriminate between bacterial infections and sterile inflammations . For this purpose, we first assessed the binding of various 99mTc-labelled natural or synthetic peptides, which are based on the sequence of the human antimicrobial peptide ubiquicidin (UBI) or human lactoferrin (hLF), to bacteria and to leucocytes in vitro . In order to select peptides that preferentially bind to bacteria over host cells, radiolabelled peptides were injected into mice intraperitoneally infected with Klebsiella pneumoniae (K . pneumoniae) and the amount of radioactivity associated with the bacteria and with the leucocytes was quantitated . The next phase focussed on discrimination between bacterial infections and sterile inflammatory processes using 99mTc-labelled peptides in mice intramuscularly infected with various bacteria (e.g . multi-drug-resistant Staphylococcus aureus) and in animals that had been injected with lipopolysaccharides (LPS) of bacterial origin to create a sterile inflammatory process . Also, we studied the distribution of 99mTc-labelled UBI 29-41 and UBI 18-35 in rabbits having an experimental thigh muscle infection with K . pneumoniae and in rabbits injected with LPS . Based on the results of our in vitro and in vivo binding assays, two peptides, i.e . UBI 29-41 and UBI 18-35, were selected as possible candidates for infection imaging . The radiolabelled peptides can detect infections with both gram-positive and gram-negative bacteria in mice as early as 5-30 min after injection, with a target-to-non-target (T/NT) ratio between 2 and 3; maximum T/NT ratios were seen within 1 h after injection . In rabbits, high T/NT ratios (>5) for 99mTc-labelled UBI 29-41 were observed from 1 h after injection . No accumulation of the selected 99mTc-labelled UBI-derived peptides was observed in thighs of mice and rabbits previously injected with LPS . Scintigraphic investigation into the biodistribution of 99mTc-labelled UBI peptides revealed that these peptides were rapidly removed from the circulation by renal excretion . Similar data were observed for 99mTc-labelled defensin 1-3 . Our data for 99mTc-labelled hLF and related peptides indicate that these compounds are less favourable for infection detection . Taken together, 99mTc-labelled UBI 18-35 and UBI 29-41 enable discrimination between bacterial infections and sterile inflammatory processes in both mice and rabbits . Based on their characteristics, we consider these peptides the candidates of preference for detection of bacterial infections in man. Nihon Kokyuki Gakkai Zasshi, 2000 Feb, 38(2), 143 - 7 {Arteriovenous fistula associated with Staphylococcus aureus sepsis in a patient with Rendu-Osler-Weber syndrome}; Ohmagari N et al.; A 58-year-old man with a history of cerebral infarction and bleeding due to duodenal ulcer wa