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| Int J Oral Maxillofac Implants, 2001 Mar-Apr, 16(2), 201 - 7 Early implant failures in patients treated with Brånemark System titanium dental implants: a retrospective study; Kronstrom M et al.; Implant failure has been associated with factors such as poor bone quality, insufficient bone volume, implant instability, unfavorable implant loading, and smoking habits . Infections and host responses may also be important factors in dental implant failure . The objectives of the present study were to identify various explanatory factors associated with titanium implant failure . Forty subjects with stage 1 non-osseointegrated titanium dental implants (NOTI) ad modum Branemark and 40 age- and gender-matched control subjects with successfully osseointegrated titanium implants (SOTI) were studied . Clinical data and gamma G immunoglobulin (IgG) antibody titers were studied . An independent t test revealed that significantly longer implants were placed in subjects with SOTI (P < .05) . Statistically significant differences in bone shape and resorption (BSR) scores were found between SOTI and NOTI (P < .05) . Logistic regression analysis identified 3 significant explanatory outcome variables: serum antibody avidity scores for Bacteroides forsythus (P < .0001), serum antibody titers to Staphylococcus aureus (P < .001), and the BSR scores (P < .05) . Antibody avidity to B forsythus and antibody titer to S aureus were therefore the 2 most important factors associated with early implant failures and with a significant predictive ability . This indicates that immunologic factors are involved in osseointegration. Eur J Biochem, 2001 May, 268(9), 2558 - 65 Human placental calreticulin characterization of domain structure and post-translational modifications; Hojrup P et al.; The domain organization and the post-translational modifications of human placenta calreticulin were analysed by MS in combination with proteolytic digestion . Prolonged treatment with trypsin, chymotrypsin, elastase, Staphylococcus aureus V8 protease, or proteinase K all led to a 6- to 7-kDa decrease in the molecular mass of calreticulin . The decrease was found to be due to cleavages in the region around residue 340 . In addition, minor fragments resulting from secondary cleavages close to the N-terminus were observed, but no stable fragments of intermediate size were found . These results show that the C-domain of calreticulin is susceptible to proteolytic cleavage and that the N- and P-domains form a proteolytically stable tight association . A disulfide bridge between the first two cysteines was mapped in the N-domain, and the third cysteine was found in the reduced form . No post-translational modifications in the form of glycosylation or phosphorylation were found . A modified form of calreticulin lacking the C-terminal hexapeptide including the KDEL endoplasmic reticulum retention sequon was isolated . Such a truncation may point to a mechanism that allows escape of calreticulin from the endoplasmic reticulum. Nippon Koshu Eisei Zasshi, 2001 Mar, 48(3), 190 - 9 {A study on the transmission of MRSA among the family members including clients of visiting nurse and related infection control}; Matsumoto K et al.; PURPOSE: The purpose of the study was to clarify MRSA (methicillin-resistant Staphylococcus aureus) transmission among clients, receiving nursing care from visiting nurse stations, and family members, as well as to determine MRSA positive rates of visiting nurses themselves and their handwashing habits . METHODS: The subjects were 131 clients who had utilized 32 visiting nurse stations, and had tested MRSA positive in our previous study performed 2-5 months earlier . The presence of MRSA in the nasal passages was investigated in 15 of the 131 MRSA positive clients and the 24 family members who had agreed to cooperate . Antibiotic sensitivity tests of the involved strains of MRSA were conducted using 14 antibiotics to allow antibiotic resistance patterns to be compared . 148 nurses who worked at 18 visiting nurse stations were also screened for MRSA in their nasal passages . In addition, a self-administered questionnaire was filled in concerning their handwashing habits . RESULTS: Out of 15 clients from whom MRSA was isolated in the previous study, 9 became MRSA positive (60.0%), and this was the case for 6 family members, living with 4 of them . The antibiotic resistance patterns coincided among the family members of 3 families, suggesting MRSA transmission among the client and his/her family member(s) . MRSA transmission was shown not to be influenced by the ADL (activities of daily living) of a client nor by the content or time of the care provided by family member(s) . As for visiting nurses, MRSA was isolated from 1 out of 148 (detection rate: 0.7%) . The practice rate for handwashing was 91.2% after visiting as compared to 22.1% before visiting, and 93.4% after care as compared to 37.5% before care; the differences were significant (P < 0.001) . The most frequently used handwashing procedures included handwashing with soap, povidone iodine and other disinfectants . The practice rate were 94.9% in visiting nurse stations and 91.2% in clients' homes . There was no significant difference between the procedures in these settings . DISCUSSION: Family members who are living with MRSA carriers are in danger of MRSA transmission irrespective of the content of the care, suggesting the need to prevent spread into compromised hosts or the community . On the other hand, visiting nurses are seldom infected with MRSA, and transmission of MRSA from nurses to clients is a rare event . This study showed, however, a low rate of handwashing before client contact . The possibility of cross infection from the hands of visiting nurses to their clients therefore needs further study. J Mol Microbiol Biotechnol, 2001 Apr, 3(2), 163 - 70 Staphylococcal multidrug efflux protein QacA; Brown MH et al.; The QacA multidrug exporter from Staphylococcus aureus mediates resistance to a wide array of monovalent or divalent cationic, lipophilic, antimicrobial compounds . QacA provides resistance to these various compounds via a proton motive force-dependent antiport mechanism that conforms to classical Michaelis-Menten kinetics . Fluorescent transport analyses have demonstrated that this QacA:substrate interaction occurs with high affinity and competition studies have shown that QacA-mediated ethidium export is competitively inhibited by other monovalent cations, and non-competitively inhibited by divalent cations, suggesting that monovalent and divalent cations bind at distinct sites on the QacA protein . The closely related export protein QacB, mediates lower levels of resistance to divalent cations, and lacks a high affinity-binding site for divalent cations . The cell membrane has been identified as the origin of QacA-mediated efflux; substrates are bound and expelled from within this hydrophobic environment . Regulation of qacA expression is achieved via the transacting repressor protein, QacR . QacR belongs to the TetR family of transcriptional repressor proteins, which all possess a helix-turn-helix DNA-binding domain at their N-terminal ends, and have highly divergent C-termini postulated to be involved in the binding of inducing compounds . QacR specifically binds to an inverted repeat, IR1, which has been identified as the qacA operator region, and overlaps the identified promoter sequence for qacA . QacR, like the multidrug export protein whose expression it regulates, has been shown to interact directly with a number of structurally-dissimilar compounds. Microbiology, 2001 May, 147(Pt 5), 1259 - 66 zur: a Zn(2+)-responsive regulatory element of Staphylococcus aureus; Lindsay JA et al.; A putative operon encoding a probable zinc-responsive regulatory element (zur) and components of an ABC-type transporter (mreA mreB) have been characterized in Staphylococcus aureus . The zur gene was inactivated but apparently this did not alter Zn(2+) uptake . Expression of mreAB zur is at a low level under a range of ion conditions . To allow inducible expression of the operon, a construct was made placing it under the control of the IPTG-inducible P(spac) promoter . Using this approach, it was shown that zur is able to repress expression of the entire operon in a Zn(2+)-dependent manner, and that mreA and mreB are likely to be involved in high-affinity ion uptake . zur has no apparent role in pathogenicity in a lesion model of S . aureus infection. Cell Biol Int, 2001, 25(4), 289 - 307 Measurement of fibroblast and bacterial detachment from biomaterials using jet impingement; Bundy KJ et al.; Fibroblast and Staphylococcus aureus detachment strength from orthopaedic alloys and a tissue culture plastic (Thermanox) have been investigated with jet impingement . For S . aureus, unlike fibroblasts, detachment is caused more by pressure than shear . For these biomaterials, detachment strength is much higher for S . aureus than fibroblasts . Comparing materials under equivalent flow conditions, S . aureus attach to stainless steel and titanium with equal strength and more strongly than to Thermanox . For fibroblasts, detachment strength from all materials was similar . Fibroblast detachment strength from these biomaterials substantially decreases with time at equal flow rates and increases with flow rate at equal exposure times . Detachment strength is very similar for 3T3 and L929 fibroblasts on Thermanox for equivalent flow rate/time combinations, though enhanced adhesion of 3T3 cells was often noted for metals . Time effects are less evident for S . aureus . S . aureus adhesion to metals is more affected by flow rate than fibroblast adhesion . Clin Infect Dis, 2001 May 15, 32(10), 1470 - 9 Epub 2001 Apr 18. Recurrent nonmenstrual toxic shock syndrome: clinical manifestations, diagnosis, and treatment; Andrews MM et al.; We report 3 cases of recurrent nonmenstrual toxic shock syndrome (TSS) and review the clinical manifestations, diagnosis, and treatment . The primary sites of infection were the genital tract (in a patient who underwent cesarean delivery), the upper respiratory tract, and a breast abscess . In all 3 patients, the initial illness was not recognized to be TSS; only after development of recurrent illness with desquamation was this diagnosis entertained . Strains of Staphylococcus aureus that were isolated from 2 patients produced TSS toxin-1, whereas the third strain produced staphylococcal enterotoxin B . All 3 patients lacked antibody to the implicated toxins at the time of presentation with recurrent illness . Nonmenstrual TSS can occur in a variety of clinical settings and may be recurrent . The presence of desquamation during a febrile, multisystem illness could suggest this diagnosis and should prompt the clinician to obtain appropriate cultures for S . aureus. Clin Infect Dis, 2001 May 15, 32(10), 1393 - 8 Epub 2001 Apr 20. Duration of colonization by methicillin-resistant Staphylococcus aureus after hospital discharge and risk factors for prolonged carriage; Scanvic A et al.; To investigate persistent carriage of methicillin-resistant Staphylococcus aureus (MRSA), we conducted a prospective 10-month study of MRSA carriage in previous carriers who were readmitted to our hospital . Four screening specimens, 2 from the skin and 2 from the nares, were obtained within 3 days after admission, in addition to diagnostic specimens requested by physicians . Of the 78 patients included in our study, 31 (40%) were persistent carriers of MRSA, with an estimated median time of 8.5 months to MRSA clearance . In the multivariate analysis, the only factor significantly associated with persistent carriage was the presence of a break in the skin at readmission (odds ratio, 4.34; P=.004); however, a trend was found for admission from a chronic-care institution (odds ratio, 3.65; P=.06) . Our data confirm that prolonged carriage of MRSA can occur after hospital discharge, support routine screening for MRSA at readmission of previously MRSA-positive patients, and suggest that a particularly high index of suspicion for MRSA carriage should be maintained if these patients have a break in the skin. Am J Ther, 2000 Sep, 7(5), 319 - 20 Vancomycin anaphylaxis in a patient with vancomycin-induced red man syndrome; Hassaballa H et al.; Vancomycin is a powerful glycopeptide antibiotic that is increasingly being used owing to the emergence of highly resistant organisms such as methicillin-resistant Staphylococcus aureus . Although a generally safe medication, administration of vancomycin is not benign, and there have been a number of adverse reactions reported . We present the case of a patient with vancomycin-induced red man syndrome who developed vancomycin anaphylaxis . Our case illustrates that red man syndrome may be a marker for true vancomycin allergy, although it was generally not thought of as so in the past. J Biol Chem, 2001 Jun 29, 276(26), 24360 - 4 Epub 2001 Apr 19. Human alveolar macrophages and granulocyte-macrophage colony-stimulating factor-induced monocyte-derived macrophages are resistant to H2O2 via their high basal and inducible levels of catalase activity; Komuro I et al.; Human alveolar macrophages (A-MPhi) and macrophages (MPhi) generated from human monocytes under the influence of granulocyte-macrophage colony-stimulating factors (GM-MPhi) express high levels of catalase activity and are highly resistant to H(2)O(2) . In contrast, MPhi generated from monocytes by macrophage colony-stimulating factors (M-MPhi) express low catalase activity and are about 50-fold more sensitive to H(2)O(2) than GM-MPhi or A-MPhi . Both A-MPhi and GM-MPhi but not M-MPhi can induce catalase expression in both protein and mRNA levels when stimulated with H(2)O(2) or zymosan . M-MPhi but not GM-MPhi produce a large amount of H(2)O(2) in response to zymosan or heat-killed Staphylococcus aureus . These findings indicate that GM-MPhi and A-MPhi but not M-MPhi are strong scavengers of H(2)O(2) via the high basal level of catalase activity and a marked ability of catalase induction and that catalase activity of MPhi is regulated by colony-stimulating factors during differentiation. Microb Pathog, 2001 Apr, 30(4), 247 - 52 Thrombin generation and mortality during Staphylococcus aureus sepsis; Bokarewa MI et al.; Sepsis-induced abnormalities of coagulation may contribute to mortality during severe bacterial infection . The aim of this study was to examine changes in coagulation parameters and to assess the role of protein C supplementation during murine S . aureus sepsis . Gram-positive sepsis was characterized by a hypercoagulable state with predominant activation of the external coagulation pathway, registered as an early increase of tissue factor activity and concomitant reduction in protein C . The internal coagulation pathway was unaffected . No correlation between the changes of coagulation parameters and the intensity of inflammation, determined as serum IL-6 levels, was found . Supplementation with neither protein C or APC favoured survival in S . aureus sepsis . Reduction in thrombin generation in response to protein C supplementation was associated with significantly increased survival . Microb Drug Resist, 2001 Spring, 7(1), 23 - 32 Molecular epidemiology of methicillin-resistant Staphylococcus aureus in Colombian hospitals: dominance of a single unique multidrug-resistant clone; Gomes AR et al.; The first study on the molecular characterization of methicillin-resistant Staphylococcus aureus (MRSA) isolates from Colombia was performed as part of a global surveillance established by the CEM/NET Initiative, under Project RESIST . Seventy-six MRSA isolates recovered from five hospitals during 1996-1998 were analyzed by the hybridization of ClaI restriction digests with mecA- and Tn554-specific probes, and by pulsed-field gel electrophoresis (PFGE) of chromosomal SmaI digests . All MRSA isolates, with one exception, belonged to a single clonal type II::NH::D . This clone, which was previously described among MRSA isolates recovered in the early 1990s in European and New York and South American hospitals, showed resistance to beta-lactam antibiotics only and appeared to be associated almost exclusively with pediatric infections ("Pediatric clone" of MRSA) . While sharing identical molecular typing properties with the Pediatric clone, the Colombian isolates differed by extensive multidrug resistance and were recovered from patients of all ages . It is also noteworthy that the Brazilian clone of MRSA (XI::B::B), another multidrug-resistant international clone currently widely spread in Brazil, Argentina, Uruguay, Chile, and also in several European countries, was completely absent from this set of isolates from Colombia. Infect Control Hosp Epidemiol, 2001 Mar, 22(3), 152 - 6 A blinded comparison of three laboratory protocols for the identification of patients colonized with methicillin-resistant Staphylococcus aureus; Gardam M et al.; OBJECTIVE: To compare three laboratory screening protocols for the detection of methicillin-resistant Staphylococcus aureus (MRSA) from surveillance specimens (mannitol-salt agar containing 2 microg/mL of oxacillin {MSA-2}, mannitol-salt agar containing 4 microg/mL of oxacillin {MSA-4}, and a broth-containing protocol as recommended by the American Society for Microbiology {M-ASM}) . DESIGN: Blinded comparative laboratory study and cost analysis . SETTING: University-affiliated microbiology laboratory . METHODS: Outcome measurements included rate of detection of MRSA-positive specimens and patients, turnaround time, and media and technologist costs . All MRSA culture swabs obtained from any patient site from November 1998 to April 1999 were included . RESULTS: The M-ASM protocol detected between 19.1% and 32.0% more MRSA-positive specimens and between 13.3% and 23.3% more MRSA-positive patients per surveillance event than the MSA-4 and MSA-2 protocols, respectively . There was no difference in positive-culture reporting time between the M-ASM and MSA4 protocols . The broth-containing protocol was 2- to 2.5-fold more expensive than the simpler protocols, taking into account media and laboratory personnel costs . CONCLUSIONS: It remains to be determined whether it is cost beneficial for a hospital to adopt the M-ASM, as the potential cost of MRSA transmission from unidentified MRSA-colonized patients is unknown . A broth-containing protocol should be considered the gold standard in future studies examining newer MRSA screening protocols Chem Pharm Bull (Tokyo), 2001 Apr, 49(4), 361 - 7 Structure-activity relationship (SAR) studies on oxazolidinone antibacterial agents . 3 . Synthesis and evaluation of 5-thiocarbamate oxazolidinones; Tokuyama R et al.; A series of 5-thiocarbamate oxazolidinones was prepared and tested for in vitro and in vivo antibacterial activities . The results of in vitro antibacterial activity indicated that the 5-thiocarbamate group was a suitable substituent for the activity by the 5-moderate hydrophilicity . The compounds within a favorable logP value range were found to have potent in vitro antibacterial activity against gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) . Compounds 3a and 4h were superior to linezolid in both in vitro and in vivo potency and were considered to be hopeful compounds . We also discuss the pharmacokinetic properties of several compounds in mice. Chem Pharm Bull (Tokyo), 2001 Apr, 49(4), 353 - 60 Structure-activity relationship (SAR) studies on oxazolidinone antibacterial agents . 2 . Relationship between lipophilicity and antibacterial activity in 5-thiocarbonyl oxazolidinones; Tokuyama R et al.; 5-Thiourea and 5-dithiocarbamate oxazolidinones were synthesized as a continuation of research on 5-thiocarbonyl oxazolidinone antibacterial agents considering the hydrophobic parameters of the molecule . The structure-activity relationship (SAR) study revealed that the antibacterial activity on 5-thiocarbonyl oxazolidinones was significantly affected by the lipophilicity, especially the calculated log P value and the balance between 5-hydrophilic (or hydrophobic) substituent and hydrophobic (or hydrophilic) substituents on the benzene ring . Some of 5-thiocarbonyl oxazolidinones were found to have good in vitro antibacterial activity against gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). J Assoc Physicians India, 2000 Nov, 48(11), 1057 - 9 Tropical pyomyositis: experience of a tertiary care hospital in north-west India; Malhotra P et al.; OBJECTIVE: Tropical pyomyositis (TP), a disease of varied aetiology, has been reported frequently from Africa and Latin America . However there are only a few reports from India . MATERIAL AND METHODS: Between January 1992 to June 1999, 26 patients with TP were admitted to Post Graduate Institute of Medical Education and Research Chandigarh, a tertiary care centre in north-west India and formed the study material . RESULTS: The disease was more common in young adults with male to female ratio being 5:2 . In majority, presenting features were muscle pains (100%) and fever (81%) . The diagnosis was confirmed by aspirating pus from the involved muscle . The pus grew organisms in 41.7% patients and methicillin sensitive Staphylococcus aureus was the commonest causative organism . Blood cultures were positive in 14.3% of cases . The common complications were bronchopneumonia (23.1%), empyema (19.2%) and venous thrombosis (15.4%) . In three patients, the disease occurred in association with tuberculosis of dorsal-lumbar spine . The overall mortality was low (7.7%) . CONCLUSIONS: TP is not an uncommon disease in India . Though the presenting features were similar to earlier studies, complications varied and mortality was low . An early recognition and aggressive treatment helps in decreasing morbidity and mortality. Mol Microbiol, 2001 Apr, 40(2), 433 - 9 Biological cost and compensatory evolution in fusidic acid-resistant Staphylococcus aureus; Nagaev I et al.; Fusidic acid resistance resulting from mutations in elongation factor G (EF-G) of Staphylococcus aureus is associated with fitness costs during growth in vivo and in vitro . In both environments, these costs can be partly or fully compensated by the acquisition of secondary intragenic mutations . Among clinical isolates of S . aureus, fusidic acid-resistant strains have been identified that carry multiple mutations in EF-G at positions similar to those shown experimentally to cause resistance and fitness compensation . This observation suggests that fitness-compensatory mutations may be an important aspect of the evolution of antibiotic resistance in the clinical environment, and may contribute to a stabilization of the resistant bacteria present in a bacterial population. Acta Anaesthesiol Scand, 2001 May, 45(5), 570 - 5 Inhibitory effects of S-(-) and R-(+) bupivacaine on neutrophil function; Welters ID et al.; BACKGROUND: Local anesthetics inhibit migration, enzyme release and superoxide anion generation of polymorphonuclear leukocytes (PMN) . Due to their ability to phagocytose and kill bacteria PMN represent a major defense mechanism in the circulating blood . In this study we determined the influence of racemic bupivacaine and its enantiomers on neutrophil phagocytic activity, oxidative burst as well as surface expression of complement and Fcgamma receptors . METHODS: Venous blood was pre-incubated with different concentrations of either racemic bupivacaine, R-(+) or S-(-) bupivacaine . Fluoresceine isothiocyanate (FITC)-labeled antibodies against Fcgamma receptor III (CD16), complement receptor 1 (CD35) and complement receptor 3 (CD11b) were used to determine surface receptor expression . Phagocytic activity was measured by ingestion of FITC-labeled vital Staphylococcus aureus . Oxidative burst was determined by conversion of nonfluorescent dihydrorhodamine 123 into fluorescent rhodamine 123 . Fluorescent intensity of each sample was determined by flow cytometry . RESULTS: Racemic bupivacaine inhibited surface receptor expression, phagocytosis, and oxidative burst in a time- and concentration-dependent manner . Although the S-(-) enantiomer exerted significantly less inhibitory action on neutrophil function compared to R-(+) and racemic bupivacaine, these effects were small compared to the overall changes . CONCLUSION: These findings suggest that bupivacaine impairs surface receptor expression and may thereby contribute to reduced phagocytic activity and oxidative burst . Enantiomer-specific effects of bupivacaine may play a minor role in the inhibition of these leukocyte functions. J Food Prot, 2001 Apr, 64(4), 546 - 50 Survival of staphylococcus aureus and Escherichia coli as affected by ethanol and NaCl; Huang SL et al.; Growth of three strains of Staphylococcus aureus and two strains of Escherichia coli on nutrient agar (NA) supplemented with ethanol and NaCl was investigated . S . aureus did not grow on NA containing > or =10% ethanol (wt/wt) combined with > or =0% NaCl (wt/wt), or 7.5% ethanol combined with 7.5% NaCl . Neither E . coli nor E . coli O157:H7 grew on NA containing > or =7.5% ethanol combined with > or =0% NaCl, 5% ethanol combined with > or =2.5% NaCl, or > or =5% NaCl combined with > or =0% ethanol . It is apparent that NaCl enhanced the inhibitory effect of ethanol on growth of S . aureus and E . coli When cells were suspended in nutrient broth containing 12.5, 20, or 40% ethanol combined with NaCl, viable cells decreased with an increase of ethanol concentration . Ethanol sensitivity among strains and between genera varied in a limited range . When the cells were exposed to 20% ethanol in combination with 5% NaCl, S . aureus and E . coli lost viability after 30 and 10 min, respectively . When treated with 40% ethanol combined with > or =0% NaCl, all test strains lost viability within 5 min. J Food Prot, 2001 Apr, 64(4), 470 - 5 Antimicrobial edible packaging based on cellulosic ethers, fatty acids, and nisin incorporation to inhibit Listeria innocua and Staphylococcus aureus; Coma V et al.; Edible cellulosic films made with hydroxypropylmethylcellulose (HPMC) have proven to be inadequate moisture barriers . To improve its water vapor barrier properties, different hydrophobic compounds were incorporated into the HPMC matrix . Some fatty acids and derivatives were included into the film-forming solution prior to film formation . Stearic acid was chosen because of its high capacity to reduce significantly the water vapor transmission rate . Antimicrobial activity of edible HPMC film was obtained by the incorporation of nisin into the film-forming solution . Nisin is an antimicrobial peptide effective against gram-positive bacteria . The inhibitory activity of this bacteriocin was tested for inhibition of Listeria innocua and Staphylococcus aureus . The use of stearic acid was observed to reduce the inhibitory activity of active HPMC film against both selected strains . This phenomenon may be explained by electrostatic interactions between the cationic nisin and the anionic stearic acid . Further studies showed that antimicrobial activity of film varied with the nature of the hydrophobic compound incorporated, in decreasing order: film without lipid, methylstearate film, and stearic acid film . This corroborated the idea of electrostatic interactions. Chemotherapy, 2001 May-Jun, 47(3), 208 - 14 Bacterial adhesiveness: effects of the SH metabolite of erdosteine (mucoactive drug) plus clarithromycin versus clarithromycin alone; Braga PC et al.; After metabolization, erdosteine (a mucoactive drug) produces an active metabolite (Met I) with an SH group that is capable of opening disulphide bonds, including those of pilin, a protein of bacterial fimbriae . This induces stereochemical conformational changes that interfere with the binding of bacterial adhesins (fimbriae) to receptors on eukaryotic cells . At subinhibitory concentrations, the macrolide clarithromycin inhibits the expression of adhesins on bacterial cell surfaces . The addition of 5 and 10 microg/ml of Met I to 1/8, 1/16 and 1/32 MIC of clarithromycin potentiated the inhibition of Staphylococcus aureus adhesiveness to human mucosal cells in comparison with the antibiotic alone . This finding opens up a new possibility of interfering with bacterial adhesiveness and its resulting pathogenicity not only by using antibiotics but also by means of their combination with agents devoid of antibacterial activity . Eur J Clin Microbiol Infect Dis, 2001 Feb, 20(2), 117 - 9 Predisposing factors and outcome of Staphylococcus aureus bacteremia in neutropenic patients with cancer; Gonzalez-Barca E et al.; Staphylococcus aureus caused 30 of 438 (7%) cases of bacteremia in neutropenic patients with cancer during a 10-year study period . Acute leukemia as an underlying disease and severe oral mucositis were more frequent among patients with Staphylococcus aureus bacteremia (57% vs . 33%, P = 0.01, and 32% vs . 12%, P = 0.006, respectively) than among the 151 patients who had gram-negative bacteremia during the same study period . The most frequent source of Staphylococcus aureus bacteremia was the venous catheter (35% vs . 1%; P = 0.00001) . Septic metastases were more frequent in patients with Staphylococcus aureus bacteremia (14% vs . 4%, P = 0.03) . Attributable mortality was 10% and overall mortality 23% . Staphylococcus aureus bacteremia remains a significant cause of morbidity and mortality in neutropenic patients with cancer. J Eur Acad Dermatol Venereol, 2000 Sep, 14(5), 393 - 6 An open study of efficacy and safety of long-term treatment with mometasone furoate fatty cream in the treatment of adult patients with atopic dermatitis; Faergemann J et al.; BACKGROUND: Atopic dermatitis is a severe chronic skin disease often deteriorated by the presence of microorganisms and often responds well to treatment with potent corticosteroids . However, the long-term use of potent topical corticosteroids are accompanied by side-effects such as skin atrophy . OBJECTIVE: To study the effect and safety of prophylactic treatment with mometasone furoate fatty cream (contains hexylene glycol) for 6 months in patients with atopic dermatitis . RESULTS: Sixty-one of 68 (90%) patients were still free of their disease after 6 months of twice weekly treatment and only one showed possible treatment related signs of skin atrophy . The number of Staphylococcus aureus and Pityrosporum ovale were significantly reduced in cleared patients . CONCLUSIONS: Mometasone furoate fatty cream is effective and safe both for treatment and as a prophylaxis in patients with atopic dermatitis. Nippon Rinsho, 2001 Apr, 59(4), 728 - 32 {Clinical relevance of hetero-VRSA in surgical infections}; Okii K et al.; Vancomycin was used increasingly for treating methicillin-resistant Staphylococcus aureus(MRSA) infection . Recently MRSA strains which showed low-level resistance to vancomycin were isolated . Vancomycin-intermediate Staphylococcus aureus(VISA) show a vancomycin minimum inhibitory concentration(MIC) of 8 micrograms/ml . VISA appear to be rare . The vancomycin resistance phenotype is reported to be unstable in such isolates . To detect heterogeneously resistant VRSA(hetero-VRSA . MIC 1 to 4 micrograms/ml), we need to use population analysis and growth on Mu3 agar plate, because MIC cannot confirm hetero-VRSA . Hetero-VRSA is not so rare(about 0-47%) . Hetero-VRSA may be responsible for failure of vancomycin therapy, but its mechanism remains unclear . Until it becomes better understood, the clinical relevance cannot be assessed. Nippon Rinsho, 2001 Apr, 59(4), 666 - 72 {Mechanisms of methicillin and vancomycin resistance in Staphylococcus aureus}; Kuroda-Murakami H; The mechanism of methicillin and vancomycin resistance in Staphylococcus aureus (MRSA) is discussed, focusing on the phenotypic expression of resistance; i.e., pre-methicillin resistance, Egle-type resistance, heterogeneous and homogeneous resistance . Acquisition of staphylococcus cassette chromosome(SCCmec) by a recipient MSSA strain in the mecA-positive, and methicillin-susceptible status(pre-MRSA status) of S . aureus . Subsequent of the mecI-gene-mediated repression of mecA gene transcription results in the heterogeneous expression of methicillin resistance . It is only with the alternation of another chromosomal locus that the heterogeneous-to-homogeneous (hetero-to-homo) conversion of methicillin resistance occurs . Acquisition of a part of vancomycin-resistance(the level of hetero-VRSA) is associated with hetero-to-homo conversion of methicillin resistance, and full expression of vancomycin resistance(MIC = 8 mg/l) is achieved by additional acceleration of cell-wall synthesis causing apparent cell-wall thickening. Neurol India, 2001 Mar, 49(1), 41 - 6 Neuropathological complications of infective endocarditis : study of autopsy material; Patel FM et al.; 78 autopsy proven cases of infective endocarditis (IE) seen during 1983 to 1995 were retrospectively reviewed . The brain was available for examination in 44 cases . In the remaining cases, brain was not examined because examination of it was not requested due to lack of neurological findings . Brain lesions were observed in 35 out of 44 cases of IE . Assuming remaining 34 cases to be without brain lesions, the brain involvement in IE would be 44.87% (35 out of 78 cases) . Mean age of all cases of IE and those with brain lesions was similar i.e . 26.5+/-16.6 years and 26.6+/-13.06 years respectively . Largest number of cases with neuropathological lesions were associated with normal valve IE (48.57%) . Mitral valve was most commonly involved in cases with CNS complications (57.14%) (p<0.05) . The various types of brain lesions were infarction (68.57%), haemorrhage (57.14%), cerebral micro-abscess (31.42%) and focal meningitis (14.28%) . More than one type of lesion was observed in 19 cases, indicating complicated nature of brain lesions in fatal cases of IE . Left sided middle cerebral artery (MCA) territory was the commonest site of infarction and haemorrhage . Staphylococcus aureus appeared to be the most common organism in fatal cases of IE . Normal valve IE with or without CNS complications constitutes a significant group in India and is different from the west as far as the predisposing conditions are concerned. Antimicrob Agents Chemother, 2001 May, 45(5), 1535 - 8 Prolonged antimicrobial activity of a catheter containing chlorhexidine-silver sulfadiazine extends protection against catheter infections in vivo; Bassetti S et al.; The present study evaluated in vitro and in vivo a new chlorhexidine (C)-silver sulfadiazine (S) vascular catheter (the CS2 catheter) characterized by a higher C content and by the extended release of the surface-bound antimicrobials . The CS2 catheter was compared with a first-generation, commercially available CS catheter (the CS1 catheter) . The CS2 catheter produced slightly smaller zones of inhibition (mean difference, 0.9 mm {P < 0.001}) at 24 h against Staphylococcus aureus and five other microorganisms by several different methodologies . However, in a rabbit model, both CS catheters were similarly efficacious in preventing a catheter infection when the rabbits were inoculated with 10(4) to 10(7) CFU of S . aureus at the time of catheter insertion . The CS2 catheter retained its antimicrobial activity significantly longer in vitro and in vivo (half-lives exceeded 34 and 7 days, respectively) and was also significantly more efficacious in preventing a catheter infection when 10(6) CFU of S . aureus was inoculated 2 days after catheter implantation (P < 0.001) . These results suggest that prolonged anti-infective activity on the external catheter surface provides improved efficacy in the prevention of infection. Antimicrob Agents Chemother, 2001 May, 45(5), 1431 - 7 Mechanism and suppression of lysostaphin resistance in oxacillin-resistant Staphylococcus aureus; Climo MW et al.; The potential for the development of resistance in oxacillin-resistant Staphylococcus aureus (ORSA) to lysostaphin, a glycylglycine endopeptidase produced by Staphylococcus simulans biovar staphylolyticus, was examined in vitro and in an in vivo model of infection . Following in vitro exposure of ORSA to subinhibitory concentrations of lysostaphin, lysostaphin-resistant mutants were idenitifed among all isolates examined . Resistance to lysostaphin was associated with a loss of resistance to beta-lactams and a change in the muropeptide interpeptide cross bridge from pentaglycine to a single glycine . Mutations in femA, the gene required for incorporation of the second and third glycines into the cross bridge, were found following PCR amplification and nucleotide sequence analysis . Complementation of lysostaphin-resistant mutants with pBBB31, which encodes femA, restored the phenotype of oxacillin resistance and lysostaphin susceptibility . Addition of beta-lactam antibiotics to lysostaphin in vitro prevented the development of lysostaphin-resistant mutants . In the rabbit model of experimental endocarditis, administration of a low dose of lysostaphin for 3 days led predictably to the appearance of lysostaphin-resistant ORSA mutants in vegetations . Coadministration of nafcillin with lysostaphin prevented the emergence of lysostaphin-resistant mutants and led to a mean reduction in aortic valve vegetation counts of 7.5 log(10) CFU/g compared to those for untreated controls and eliminated the isolation of lysostaphin-resistant mutants from aortic valve vegetations . Treatment with nafcillin and lysostaphin given alone led to mean reductions of 1.35 and 1.65 log(10) CFU/g respectively . In ORSA, resistance to lysostaphin was associated with mutations in femA, but resistance could be suppressed by the coadministration of beta-lactam antibiotics. Antimicrob Agents Chemother, 2001 May, 45(5), 1323 - 36 Structural comparison of three types of staphylococcal cassette chromosome mec integrated in the chromosome in methicillin-resistant Staphylococcus aureus; Ito T et al.; The beta-lactam resistance gene mecA of Staphylococcus aureus is carried by a novel mobile genetic element, designated staphylococcal cassette chromosome mec (SCCmec), identified in the chromosome of a Japanese methicillin-resistant S . aureus (MRSA) strain . We now report identification of two additional types of mecA-carrying genetic elements found in the MRSA strains isolated in other countries of the world . There were substantial differences in the size and nucleotide sequences between the elements and the SCCmec . However, new elements shared the chromosomal integration site with the SCCmec . Structural analysis of the new elements revealed that they possessed all of the salient features of the SCCmec: conserved terminal inverted repeats and direct repeats at the integration junction points, conserved genetic organization around the mecA gene, and the presence of cassette chromosome recombinase (ccr) genes responsible for the movements of SCCmec . The elements, therefore, were considered to comprise the SCCmec family of staphylococcal mobile genetic elements together with the previously identified SCCmec . Among 38 epidemic MRSA strains isolated in 20 countries, 34 were shown to possess one of the three typical SCCmec elements on the chromosome . Our findings indicated that there are at least three distinct MRSA clones in the world with different types of SCCmec in their chromosome. J Am Geriatr Soc, 2001 Mar, 49(3), 270 - 6 Colonization of skilled-care facility residents with antimicrobial-resistant pathogens; Trick WE et al.; OBJECTIVES: To determine the frequency of and risk factors for colonization of skilled-care unit residents by several antimicrobial-resistant bacterial species, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococcus (VRE), or extended-spectrum-beta-lactamase-producing (ESBL-producing) (ceftazidime resistant) Klebsiella pneumoniae or Escherichia coli . DESIGN: Point-prevalence survey and medical record review . SETTING: The skilled-care units in one healthcare facility . PARTICIPANTS: 120 skilled-care unit residents . MEASUREMENTS: Colonization by each of the four antimicrobial-resistant pathogens during a point-prevalence survey, using rectal, nasal, gastrostomy-tube site, wound, and axillary cultures, June 1-3, 1998; 117 (98%) had at least one swab collected and 114 (95%) had a rectal swab collected . Demographic and clinical characteristics were evaluated as risk factors for colonization . All isolates were strain typed by pulsed-field gel electrophoresis of total genomic deoxyribonucleic acid . RESULTS: Of 117 participants, 50 (43%) were culture positive for > or =1 antimicrobial-resistant pathogen: MRSA (24%), ESBL-producing K . pneumoniae (18%) or E . coli (15%), and VRE (3.5%) . Of 50 residents culture positive for any of these four antimicrobial-resistant species, 13 (26%) were colonized by more than one resistant species; only three (6%) were on contact-isolation precautions at the time of the prevalence survey . Risk factors for colonization varied by pathogen: total dependence on healthcare workers (HCWs) for activities of daily living (ADLs) and antimicrobial receipt for MRSA, total dependence on HCWs for ADLs for ESBL-producing K . pneumoniae, and antimicrobial receipt for VRE . No significant risk factors were identified for colonization by ESBL-producing E . coli . Among colonized patients, there was a limited number of strain types for MRSA (24 patients, 4 strain types) and ESBL-producing K . pneumoniae (21 patients, 3 strain types), and a high proportion of unique strain types for VRE (4 patients, 4 strain types) and FSBL-producing E . coli (17 patients, 10 strain types) . CONCLUSION: A large unrecognized reservoir of skilled-care-unit residents was colonized by antimicrobial-resistant pathogens, and co-colonization by more than one target species was common . To prevent transmission of antimicrobial-resistant pathogens in long-term care facilities in which residents have high rates of colonization, infection-control strategies may need to be modified . Potential modifications include enhanced infection-control strategies, such as universal gloving for all or high-risk residents, or screening of high-risk residents, such as those with total dependence on HCWs for ADLs or recent antimicrobial receipt, and initiation of contact-isolation precautions for colonized residents. J Intern Med, 2001 Apr, 249(4), 289 - 95 Specific T-cell receptor usage with cytokinemia in Henoch-Schönlein purpura nephritis associated with Staphylococcus aureus infection; Hirayama K et al.; OBJECTIVES: We sought to evaluate the mechanism of Henoch-Schonlein purpura nephritis (HSPN) associated with Staphylococcus aureus (S . aureus) infection . DESIGN: We evaluated six male patients with HSPN associated with S . aureus infection . Routine laboratory examinations, bacteriological examination, histological examination, and analysis of serum cytokine levels were performed in all cases . In addition, peripheral blood mononuclear cells (PBMC) obtained from the six patients and 45 normal individuals were stained with labelled-monoclonal antibodies against six variable parts of the beta-chain (Vbeta) of the T-cell receptor (TCR), and stained cells were analysed by flow cytometry . RESULTS: Patients with HSPN associated with S . aureus infection showed features of the nephrotic syndrome with rapidly progressive glomerulonephritis, as well as varying degrees of mesangial proliferative glomerulonephritis with crescent formation . Serological examination showed elevated levels of serum IgA and IgG as well as immune complexes after the onset of infection . The percentage of Vbeta-(5.2 + 5.3) and Vbeta 8-positive cells in patients with HSPN were significantly higher than in normal individuals; moreover, specific TCR-Vbeta usage was not observed in patients with HSPN whose S . aureus infection had improved . Serum levels of IL-1beta, IL-2, IL-6, IL-8 and TNF-alpha in patients with HSPN were significantly higher than in normal individuals, and normalized at the healing stage of S . aureus infection . CONCLUSION: Conventional antigens and/or staphylococcal enterotoxins originated from S . aureus might have been involved in the pathogenesis of HSPN in the present cohort . Therefore, steroid or other immunosuppressive therapies could not be utilized despite the high activity of glomerulonephritis, and as a result the prognoses of these cases of HSPN were serious. Clin Microbiol Infect, 2001 Feb, 7(2), 65 - 9 Uptake and intracellular activity of ketolide HMR 3647 in human phagocytic and non-phagocytic cells; Pascual A et al.; OBJECTIVE: To evaluate the uptake of HMR 3647 into human neutrophils (PMNs), human peritoneal macrophages (PMOs) and tissue-cultured cells (epithelial cells and fibroblasts), and to assess the intracellular activity of this drug . METHOD: Cell uptake of HMR 3647 was measured by radiometric assay, as described by Klemper and Styrt . Intracellular activity was determined by incubation for 3 h of PMNs containing bacteria in the presence of HMR 3647 . RESULTS: The intracellular concentrations were 130 and 71 times higher than extracellular concentrations in PMNs and PMOs, respectively (extracellular concentrations: 2-25 mg/L) . The cellular-to-extracellular concentration ratios (C/E) for tissue-cultured cells were lower than those obtained in phagocytic cells but still greater than 5 . The uptake of HMR 3647 was rapid and non-saturable in all cells . HMR 3647 was released slowly from phagocytic cells . HMR 3647 (extracellular concentration: 0.5-10 mg/L) did not significantly reduce the intracellular survival rate of Staphylococcus aureus ATCC 25923 in PMNs . CONCLUSIONS: HMR 3647 reaches intracellular concentrations several times higher than extracellular concentrations within phagocytic and non-phagocytic cells . The slow efflux of this drug from phagocytic cells suggests that these cells may be a vehicle for it, delivering it to sites of infection. Microbiol Res, 2001 Mar, 155(4), 345 - 9 Isolation and characterization of methicillin-resistant Staphylococcus aureus strains from nares of nurses and their gowns; Horikawa K et al.; The isolation and characterization of methicillin-resistant Staphylococcus aures (MRSA) strains from the bilateral nares of nurses and their gowns are described . MRSA strains could be isolated from eigth of fifty bilateral nares of nurses and two of their gowns . Ten MRSA strains were typed using coagulase typing, and divided into two types, coagulase II and III . In this study, we found a new group (producing toxic shock syndrome toxin -1, coagulase III and staphylococcal enterotoxin C) in Japanese MRSA . Furthermore, we confirmed that MRSA strains originating from bilateral nares of three nurses were identical and two strains isolated from the left naris of one nurse and her gown were also identical by pulsed-field gel electrophoresis. Microbiol Res, 2001 Mar, 155(4), 339 - 44 Characterization of hemolytic activity of Staphylococcus aureus strains isolated from bovine mastitic milk; Ali-Vehmas T et al.; Beta (beta) and delta (delta)-hemolysin of Staphylococcus aureus strains were cultured in vitro in milk lactoserum (whey) prepared from both healthy and mastitis bovine milk . Production of beta- and delta-hemolysins were detected in 12 out of 50 strains studied . The association between N-acetyl-beta-D-glucosaminidase (NAGase) activity, plasmin activity (PL) and trypsin inhibitory capacity (TIC), known as inflammatory indicators for mastitis, and hemolytic activity were also studied . Mastitic milk decreased directly the lytic effect of both beta-and delta-hemolysins of S . aureus on hemolytical blood agar plates . S . aureus in healthy milk samples produced more beta-hemolysin (3 times) and delta-hemolysin (2 times) when compared to S . aureus supernatants in milk from infected quarters . Furthermore, beta- and delta-hemolysis correlated negatively with TIC and NAGase and PL activities . Addition of reduced glutathione (GSH) or beta-mercaptoethanol into the artificial medium enhanced hemolysins activity. J Allergy Clin Immunol, 2001 Apr, 107(4), 607 - 14 Total and specific IgE in nasal polyps is related to local eosinophilic inflammation; Bachert C et al.; BACKGROUND: Nasal polyps (NPs) are characterized by eosinophilic inflammation and often coexist with asthma . However, the role of atopy and IgE in NP pathogenesis is unclear . OBJECTIVE: We sought to determine whether there is an association between total and specific IgE to a variety of allergens in polyp and nonpolyp tissue and markers of eosinophilic inflammation or skin test results . METHODS: Homogenates were prepared from nasal tissue of 20 patients with NPs and 20 patients without NPs and analyzed for concentrations of IL-5, IL-4, eotaxin, leukotriene (LT) C4/D4/E4, sCD23, and histamine (ELISA) . Eosinophil cationic protein (ECP), tryptase, and total and specific IgE for inhalant allergens and Staphylococcus aureus enterotoxins were measured (ImmunoCAP) . RESULTS: The concentrations of total IgE, IL-5, eotaxin, ECP, LTC4/D4/E4, and sCD23 were significantly higher in NP tissue compared with nonpolyp tissue . Total IgE was significantly correlated to IL-5, ECP, LTC4/D4/E4, and sCD23 and to the number of eosinophils in NPs . On the basis of the presence of specific IgE antibodies in tissue, 3 NP groups were defined . NP group 1 demonstrated no measurable specific IgE, and NP group 2 selected specific IgE . The third group demonstrated a multiclonal specific IgE, including IgE to S aureus enterotoxins, a high total IgE level, and a high prevalence of asthma . CONCLUSIONS: These studies suggest that there is an association between increased levels of total IgE, specific IgE, and eosinophilic inflammation in NPs, which may be of relevance in the pathophysiology of nasal polyposis . Similarly, the presence of specific IgE to staphylococcal enterotoxins A and B also points to a possible role of bacterial superantigens. J Heart Lung Transplant, 2001 Apr, 20(4), 483 - 5 Bacterial endocarditis: a rare complication following orthotopic cardiac transplantation; Kunst H et al.; We describe a 46-year-old man who developed infective endocarditis, meningitis, sternal abscess, and infective cerebral emboli after cardiac transplantation . Staphylococcus aureus was the infective organism . We successfully managed the patient with flucloxacillin and fusidic acid to treat infection, and with prostacyclin for systemic embolization. Int J Antimicrob Agents, 2001 Apr, 17(4), 327 - 9, discussion 329-30 Staphylococcus aureus bacteriuria and surgical site infections by methicillin-resistant Staphylococcus aureus; Matsukawa M et al.; Surgical site infection (SSI) remains an important cause of morbidity among hospitalized patients . We reviewed 421 patients who underwent open urological operations between January 1993 and December 1997 in our institute . Group I consisted of 259 patients who received uncontrolled antimicrobial prophylaxis (AMP) between 1993 and 1995 . Group II consisted of 162 patients who received controlled AMP between 1996 and 1997 . In group II, penicillins or first to second-generation cephalosporins was used and the duration of use for these agents regulated according to the wound class of each operation . The operations with clean wounds showed the lowest rate of SSI in both groups; the operations with contaminated wounds showed the highest rate of SSI (32.0% in group I and 33.3% in group II) . There was no significant difference in the total rates of SSI between the two groups (P=0.216) . The most frequently isolated bacterial species was methicillin-resistant Staphylococcus aureus (MRSA), isolated in 73.3% of the cases in group I and in 93.3% in group II . There was no significant difference in the incidence of MRSA isolation between the two groups (P=0.114) . The controlled AMP could not lower the incidence of MRSA-induced SSIs . In SSI patients, 22.7% of group I and 35.7% in group II, had MRSA bacteriuria before operation . The prohibition of third-generation cephalosporins and shorter duration of AMP did not reduce the incidence of SSI induced by MRSA because MRSA was not the emerging microorganism but rather a resident in the urological ward . On the other hand, the total incidence of SSI did not increase after regulation of AMP . This finding suggests that older antibacterial agents can prevent infection, except those caused by resistant microorganisms such as MRSA . The effective counter-measure for the prevention of MRSA-induced SSI is needed. Vet Microbiol, 2001 May 21, 80(2), 131 - 8 Staphylococcus aureus associated with mammary glands of cows: genotyping to distinguish different strains among herds; Joo YS et al.; The hypothesis that strains of Staphylococcus aureus are more likely to be unique to a herd than common to several herds was tested . Herds (n=28) from nine geographic areas of Korea, with elevated milk somatic cell counts (>500000 cells/ml) were enrolled in this study . Mammary quarter milk samples were aseptically collected from all lactating cows (n=616) with at least three functional quarters . Milk was cultured and S . aureus isolates were typed using pulse field gel electrophoresis of DNA SmaI digests . A total of 181 cows were identified as having S . aureus intramammary infections . A total of 52 different types of S . aureus were identified and 34 (65.4%) were associated with a single herd . A total of 18 types of S . aureus were found in multiple herds; 14 types were found in two herds, and four types were found in three herds . Herds with 1, 2, 3, and more than 3 types, were: four (14.3%); eight (28.6%); nine (32.1%); and seven (25.0%) . The data indicate that the majority of strains were found in one herd only, and more than 90% were found in two or less herds, suggesting that strains of S . aureus are more likely to be restricted to a single herd, than found in multiple herds. J Biol Chem, 2001 Jun 29, 276(26), 24015 - 22 Epub 2001 Apr 09. Structure and mechanism of action of an indolicidin peptide derivative with improved activity against gram-positive bacteria; Friedrich CL et al.; Indolicidin, an antimicrobial peptide with a unique amino acid sequence (ILPWKWPWWPWRR-NH(2)) is found in bovine neutrophils . A derivative of indolicidin, CP10A, has alanine residues substituted for proline residues and has improved activity against Gram-positive organisms . Transmission electron microscopy of Staphylococcus aureus and Staphylococcus epidermidis treated with CP10A showed mesosome-like structures in the cytoplasm . The peptide at 2-fold the minimal inhibitory concentration did not show significant killing of S . aureus ISP67 (a histidine, uridine, and thymidine auxotroph) but did show an early effect on histidine and uridine incorporation and, later, an effect on thymidine incorporation . Upon interaction with liposomes, detergents, and lipoteichoic acid, CP10A was shown by circular dichroism spectroscopy to undergo a change in secondary structure . Fluorescence spectroscopy indicated that the tryptophan residues were located at the hydrophobic/hydrophilic interface of liposomes and detergent micelles and were inaccessible to the aqueous quencher KI . The three-dimensional structure of CP10A in the lipid mimetic dodecylphosphocholine was determined using two-dimensional NMR methods and was characterized as a short, amphipathic helical structure, whereas indolicidin was previously shown to have an extended structure . These studies have introduced a cationic peptide with a unique structure and an ability to interact with membranes and to affect intracellular synthesis of proteins, RNA, and DNA. Emerg Infect Dis, 2001 Mar-Apr, 7(2), 327 - 32 Increasing resistance to vancomycin and other glycopeptides in Staphylococcus aureus; Tenover FC et al.; Strains of Staphylococcus aureus with reduced susceptibility to glycopeptides have been reported from Japan, the United States, Europe, and the Far East . Although isolates with homogeneous resistance to vancomycin (MICs = 8 microg/mL) continue to be rare, there are increasing reports of strains showing heteroresistance, often with vancomycin MICs in the 1-4 microg/mL range . Most isolates with reduced susceptibility to vancomycin appear to have developed from preexisting methicillin-resistant S . aureus infections . Many of the isolates with reduced susceptibility to glycopeptides have been associated with therapeutic failures with vancomycin . Although nosocomial spread of the vancomycin-intermediate S . aureus (VISA) strains has not been observed in U.S . hospitals, spread of VISA strains has apparently occurred in Japan . Broth microdilution tests held a full 24 hours are optimal for detecting resistance in the laboratory; however, methods for detecting heteroresistant strains are still in flux . Disk-diffusion tests, including the Stokes method, do not detect VISA strains . The Centers for Disease Control and Prevention and other groups have issued recommendations regarding appropriate infection control procedures for patients infected with these strains. Emerg Infect Dis, 2001 Mar-Apr, 7(2), 323 - 6 Molecular epidemiology of methicillin-resistant Staphylococcus aureus; Shopsin B et al.; Subtyping methicillin- resistant Staphylococcus aureus (MRSA) isolates and tracking nosocomial infections have evolved from phenotypic to genotypic approaches; most laboratories now depend on pulsed-field gel electrophoresis (PFGE) . We discuss the limitations of current image-based genotyping methods, including PFGE, and the advantages (including ease of entering data into a database) of using DNA sequence analysis to control MRSA infections in health-care facilities. Emerg Infect Dis, 2001 Mar-Apr, 7(2), 178 - 82 The changing epidemiology of Staphylococcus aureus? Chambers HF. Strains of methicillin-resistant Staphylococcus aureus (MRSA), which had been largely confined to hospitals and long-term care facilities, are emerging in the community . The changing epidemiology of MRSA bears striking similarity to the emergence of penicillinase-mediated resistance in S . aureus decades ago . Even though the origin (hospital or the community) of the emerging MRSA strains is not known, the prevalence of these strains in the community seems likely to increase substantially. Bioorg Med Chem Lett, 2001 Apr 9, 11(7), 919 - 21 Synthesis and biological evaluation of benzo{b}naphthyridones, a series of new topical antibacterial agents; Tabart M et al.; We describe here the synthesis and biological evaluation of a series of benzo{b}naphthyridones, a new family of tricyclic antibacterial compounds that have a gram-positive spectrum of activity . RP60556A, one of the most potent of these compounds, is bactericidal against multiresistant cocci, especially multiresistant Staphylococcus aureus strains . Its physico-chemical and biological properties make it particularly suitable for topical antibacterial use. Acta Microbiol Pol, 2000, 49(3-4), 237 - 42 Effect of Staphylococcus aureus serine proteinase on the respiratory burst in phagocytic cells in vitro; Miedzobrodzki J et al.; The in vitro effects of the Staphylococcus aureus serine proteinase (SASP) on the respiratory burst of human blood mononuclear phagocytes and rat lung macrophages were investigated . The generation of reactive oxygen species (ROS), determined by means of luminol-based chemiluminescence, was stimulated by treatment with SASP in both types of the defense cells . Cell activation depended on the concentration of the enzyme and the response of monocytes was an order of magnitude stronger relative to macrophages . The chemiluminescence emission kinetics were different for both cell types and the maximum signal was achieved in approximately 3 and 17 min, respectively . In experiments involving further cell activation by latex particles, macrophages pretreated with various SASP concentrations reacted with enhanced ROS generation whereas for monocytes, the latex-induced chemiluminescence was weakened by the enzyme . The results concerning the modification of the phagocytic host cell activity by SASP suggest that this enzyme might play an important role in pathomechanisms of staphylococcal infections in vivo. Acta Pol Pharm, 2000 Nov, 57 Suppl, 117 - 9 Antimicrobial activity of selected non-antibiotics--activity of methotrexate against Staphylococcus aureus strains; Kruszewska H et al.; Some non-antibiotic drugs, e.g . cytostatics, anaesthetics or vasodilators may also show the antimicrobial activity . The aim of this study was to detect and characterise the antimicrobial activity expressed by selected non-antibiotic drugs analysed during state control performed in the Drug Institute . Over 60 drugs were randomly chosen from different groups . The surveillance study was performed on standard microbial strains: S . aureus ATCC 6538P, E . coli ATCC 8739, P . aeruginosa ATCC 15442 and C . albicans ATCC 10231 . It was found that the drugs listed below inhibited growth at least one of the examined strains: Acesan 0.075 tabl., Benuron 500 mg tabl., Chlorchex 0.5 aerosol, Methotrexat-Ebewe 500 mg amp., Naproxen 500 mg tabl., Nospa Forte 60 mg tabl., Platamine 50 mg amp., Platidiam 50 mg amp., Sensit 50 mg drag., Septofervex 2 mg tabl., Seractil 400 mg tabl., Sermion 4 mg amp., Sinemet 125 mg tabl., Tarproxen 500 mg tabl . and Zyban 150 mg tabl . It was interesting, that strong antimicrobial activity of methotrexate was limited to Staphylococcus aureus strain . Minimal inhibitory concentration of methotrexate was determined by agar dilution method with the use of 54 clinical S . aureus strains (MRSA-32 and MSSA-22) . For MSSA strains, MIC50 and MIC90 were 10 micrograms/ml and 20 micrograms/ml, respectively (range: 10-20 micrograms/ml) . For MRSA strains, MIC50 and MIC90 were 20 micrograms/ml and 100-> 100 micrograms/ml, respectively (range: 10-< 100 micrograms/ml). Crit Care Med, 2001 Apr, 29(4 Suppl), N128 - 34 Infection control measures to limit antimicrobial resistance; Warren DK et al.; Increasing antimicrobial resistance has resulted in a rapidly decreasing array of therapeutic options for infections in the critical care setting . Reports of reduced susceptibility to vancomycin in Staphylococcus aureus raise the possibility of patients being infected with a virulent pathogen for which most antibiotics are ineffective . Infection control methods to contain resistance, exclusive of antimicrobial restrictions, focus on surveillance to identify carriers of resistant organisms, prevention of nosocomial infections, adequate hand hygiene, isolation of patients who harbor resistant organisms, and the use of barrier techniques such as gowns and gloves . Surveillance using clinical isolates alone is inadequate for the identification of the majority of patients who carry resistant organisms . However, it is unclear what intensity of surveillance is needed to control the spread of these organisms in the intensive care unit in nonoutbreak situations . Attempts at eradicating carriage are often unsuccessful when there is extranasal colonization with methicillin-resistant S . aureus . Transmission of resistant organisms is primarily the result of transient contamination of healthcare workers' hands . Adequate handwashing, isolation of carriers, and barrier techniques are all necessary for containing resistance within the intensive care unit, however, compliance with these measures can be compromised by high staff turnover and heavy workload. Crit Care Med, 2001 Apr, 29(4 Suppl), N100 - 7 Antimicrobial management strategies for Gram-positive bacterial resistance in the intensive care unit; Schentag JJ; This article summarizes the current situation with Gram-positive infections, including the two primary consequences-failure to cure and resistance-relevant to the intensive care unit . The past few years have seen Enterococcus faecium resistance to vancomycin increase from 10% of strains to approaching 60% of strains in some centers . Failure is now so frequent that vancomycin can no longer be safely used . This has lead to use of two new antibiotics, quinupristin/dalfopristin (Synercid), first marketed in the United States in September 1999, and linezolid (Zyvox), which reached the U.S . market in May 2000 . Both of these agents are being used to treat culture-proven vancomycin-resistant E . faecium . The calculated areas under the inhibitory curve (AUIC) values of vancomycin, even when its minimal inhibitory concentration (MIC) is 4.0 microg/mL, show almost all vancomycin-resistant E . faecium have AUICs <125 . This explains failure, as well as the further selection of this bacteria into subpopulations with progressively higher MICs . Less well defined, but potentially an even greater problem, is the poor efficacy of vancomycin against multiresistant Staphylococcus aureus . Here, there is evidence of clinical failure in lower respiratory tract infection patients, but in most cases the MIC values of the organism have not risen to the point where AUICs are <125 . However, the minimum bactericidal concentration of this organism may be considerably higher than its MIC, and in other cases there may be a high inoculum effect or a protein-binding effect to explain the failure of vancomycin to kill multiresistant S . aureus . Besides the increasing use of the new agents, strategies to manage these two increasingly resistant Gram-positive infections include cephalosporin restriction, switch and streamlining when cultures come back from the lab, combination regimens, and cycling in selected intensive care units. Crit Care Med, 2001 Apr, 29(4 Suppl), N92 - 6 Evolving antimicrobial chemotherapy for Staphylococcus aureus infections: Our backs to the wall; Daum RS et al.; Staphylococcus aureus is responsible for many nosocomial and community-acquired infections . Its evolving resistance to traditional antimicrobial chemotherapy and emerging prevalence outside of the healthcare environment are serious concerns . This review of the changing epidemiology of methicillin-resistant S . aureus, the emergence of vancomycin (glycopeptide)-resistant isolates, and the mechanisms of resistance to beta-lactams and glycopeptides provides an update for clinicians regarding effective strategies for treatment. Crit Care Med, 2001 Apr, 29(4 Suppl), N82 - 6 Impact of Gram-positive resistance on outcome of nosocomial pneumonia; Bodi M et al.; Among Gram-positive pathogens, Staphylococcus aureus is the leading cause of death from nosocomial pneumonia . The bacterium developed progressive resistance to beta-lactams, and methicillin-resistant strains emerged in the 1980s . In consequence, vancomycin has become the drug of choice for treatment of this infection over the last decade, based on susceptibility tests and the serum antimicrobial levels recorded . However, half of the patients treated with vancomycin have died . In contrast, in patients receiving beta-lactams for pneumonia caused by methicillin-sensitive S . aureus, survival is the rule . These observations, together with the emergence of isolates with reduced susceptibility to glycopeptides, raised concern about the use of vancomycin as standard therapy for pneumonia caused by Gram-positive cocci . Maintaining tissue levels above minimal inhibitory concentration is vital to successful clinical outcome . Optimizing treatment focusing on this goal and new antimicrobials provide new opportunities to improve survival . (Crit Care Med 2001; 29{Suppl.}:N82-N86) Science, 2001 Apr 6, 292(5514), 114 - 6 A link between virulence and ecological abundance in natural populations of Staphylococcus aureus; Day NP et al.; Staphylococcus aureus is a major cause of severe infection in humans and yet is carried without symptoms by a large proportion of the population . We used multilocus sequence typing to characterize isolates of S . aureus recovered from asymptomatic nasal carriage and from episodes of severe disease within a defined population . We identified a number of frequently carried genotypes that were disproportionately common as causes of disease, even taking into account their relative abundance among carriage isolates . The existence of these ecologically abundant hypervirulent clones suggests that factors promoting the ecological fitness of this important pathogen also increase its virulence. Infect Immun, 2001 May, 69(5), 3120 - 7 Clumping factor A mediates binding of Staphylococcus aureus to human platelets; Siboo IR et al.; The direct binding of bacteria to platelets may be an important virulence mechanism in the pathogenesis of infective endocarditis . We have previously described Staphylococcus aureus strain PS12, a Tn551-derived mutant of strain ISP479, with reduced ability to bind human platelets in vitro . When tested in an animal model of endocarditis, the PS12 strain was less virulent than its parental strain, as measured by bacterial densities in endocardial vegetations and incidence of systemic embolization . We have now characterized the gene disrupted in PS12 and its function in platelet binding . DNA sequencing, Southern blotting, and PCR analysis indicate that PS12 contained two Tn551 insertions within the clumping factor A (ClfA) locus (clfA) . The first copy was upstream from the clfA start codon and appeared to have no effect on ClfA production . The second insertion was within the region encoding the serine aspartate repeat of ClfA and resulted in the production of a truncated ClfA protein that was secreted from the cell . A purified, recombinant form of the ClfA A region, encompassing amino acids 40 through 559, significantly reduced the binding of ISP479C to human platelets by 44% (P = 0.0001) . Immunoprecipitation of recombinant ClfA that had been incubated with solubilized platelet membranes coprecipitated a 118-kDa platelet membrane protein . This protein does not appear to be glycoprotein IIb . These results indicate that platelet binding by S . aureus is mediated in part by the direct binding of ClfA to a novel 118-kDa platelet membrane receptor. Infect Immun, 2001 May, 69(5), 2996 - 3003 Subinhibitory clindamycin differentially inhibits transcription of exoprotein genes in Staphylococcus aureus; Herbert S et al.; It has long been known that certain antibiotics, at subinhibitory concentrations, differentially inhibit the synthesis of alpha-hemolysin and other staphylococcal virulence factors . In this report, we show that subinhibitory clindamycin (SBCL) eliminates production of nearly all exoproteins by Staphylococcus aureus but has virtually no effect on cytoplasmic proteins . The effect was abolished by a gene conferring resistance to macrolides-lincosamides-streptogramin B, showing that differential inhibition of protein synthesis is responsible; remarkably, however, subinhibitory clindamycin blocked production of several of the individual exoprotein genes, including spa (encoding protein A), hla (encoding alpha-hemolysin), and spr (encoding serine protease), at the level of transcription, suggesting that the primary effect must be differential inhibition of the synthesis of one or more regulatory proteins . In contrast to earlier reports, however, we found that subinhibitory clindamycin stimulates synthesis of coagulase and fibronectin binding protein B, also at the level of transcription . agr and sar expression was minimally affected by subinhibitory clindamycin . These effects varied from strain to strain and do not seem to be responsible for the effects of subinhibitory clindamycin on the overall exoprotein pattern. Infect Immun, 2001 May, 69(5), 2872 - 7 Staphylococcus aureus fibronectin binding proteins are essential for internalization by osteoblasts but do not account for differences in intracellular levels of bacteria; Ahmed S et al.; Staphylococcus aureus is a major pathogen of bone that has been shown to be internalized by osteoblasts via a receptor-mediated pathway . Here we report that there are strain-dependent differences in the uptake of S . aureus by osteoblasts . An S . aureus septic arthritis isolate, LS-1, was internalized some 10-fold more than the laboratory strain 8325-4 . Disruption of the genes for the fibronectin binding proteins in these two strains of S . aureus blocked their ability to be internalized by osteoblasts, thereby demonstrating the essentiality of these genes in this process . However, there were no differences in the capacity of these two strains to bind to fibronectin or osteoblasts . Analysis of the kinetics of internalization of the two strains by osteoblasts revealed that strain 8325-4 was internalized only over a short period of time (2 h) and to low numbers, while LS-1 was taken up by osteoblasts in large numbers for over 3 h . These differences in the kinetics of uptake explain the fact that the two strains of S . aureus are internalized by osteoblasts to different extents and suggest that in addition to the fibronectin binding proteins there are other, as yet undetermined virulence factors that play a role in the internalization process. Eur Respir J, 2000 Dec, 16(6), 1152 - 7 Ventilator associated pneumonia: quality of nonbronchoscopic bronchoalveolar lavage sample affects diagnostic yield; Baughman RP et al.; The importance of predefined criteria for acceptable samples of respiratory therapists obtained lower respiratory samples were studied, using a nonbronchoscopic bronchoalveolar lavage (BAL) protocol for ventilated patients in the intensive care unit . Therapists were instructed and asked to follow guidelines for obtaining samples . Over one year, 219 samples were obtained by respiratory therapists . Of these, 115 were considered to be adequate samples using the following criteria: 60 mL of instilled volume, at least 5 mL of fluid aspirated, specimens sent for semiquantitative culture, a differential cell count of <5% bronchial epithelial cells . Overall, 52 samples grew one or more pathogen at >10,000 colony forming units (cfu).mL(-1) of BAL . The most common pathogen was Staphylococcus aureus (S . aureus) (11 samples), although Gram-negative bacilli were the single pathogen in 21 specimens . Of the 115 acceptable samples, 40 (35%) grew > or =1 pathogen at >10,000 cfu.mL(-1) . For the 80 not acceptable samples which were sent for appropriate culture, 12 (15%) grew >10,000 cfu.mL(-1) BAL . This difference was significant (Chi-squared=9.44, p<0.01) . Nonbronchoscopic bronchoalveolar lavage can be safely performed by respiratory therapists' . The authors recommend that a protocol be used to evaluate the quality of a bronchoalveolar lavage sample in the same manner sputum samples are screened prior to interpretation. J Immunol, 2001 Apr 15, 166(8), 5129 - 38 Fibronectin binding protein A of Staphylococcus aureus can mediate human T lymphocyte adhesion and coactivation; Miyamoto YJ et al.; The extracellular matrix protein fibronectin (FN) mediates the adhesion of bacteria as well as T lymphocytes . Mammalian cells express integrins alpha(4)beta(1) and alpha(5)beta(1) as the major FN-binding cell surface receptors . Bacteria such as Staphylococcus aureus, also express FN-binding receptors that are important for adherence to host tissue and initiation of infection . The S . aureus FN-binding protein, FnbpA, has been previously identified, and recombinant proteins that correspond to distinct functional regions of this protein have been made . Three recombinant truncated forms of FnbpA, rFnbpA(37-881), rFnbpA(37-605), and rFnbpA(620-881), were examined for effects on in vitro adhesion and coactivation of human T lymphocytes . These proteins, when coimmobilized with anti-CD3 mAb, activated T lymphocyte proliferation . The coactivation signal generated by the rFnbpA proteins required medium containing serum with FN . Furthermore, the costimulatory signal could be restored in FN-depleted serum when the rFnbpAs were preloaded with soluble FN . Monoclonal Ab blocking studies revealed that integrin alpha(5)beta(1) is the major receptor responsible for the rFnbpA costimulatory signal . Shear flow cell detachment assays confirmed that lymphocytes can bind to FN captured by the rFnbpA proteins . These results suggest that the S . aureus rFnbpA can interact with integrin alpha(5)beta(1) via an FN bridge to mediate adhesion and costimulatory signals to T lymphocytes. J Immunol, 2001 Apr 15, 166(8), 5108 - 14 LOX-1 supports adhesion of Gram-positive and Gram-negative bacteria; Shimaoka T et al.; Adhesion of bacteria to vascular endothelial cells as well as mucosal cells and epithelial cells appears to be one of the initial steps in the process of bacterial infection, including infective endocarditis . We examined whether lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1), a member of scavenger receptor family molecules with C-type lectin-like structure, can support adhesion of Gram-positive and Gram-negative bacteria . Chinese hamster ovary-K1 (CHO-K1) cells stably expressing LOX-1 can support binding of FITC-labeled Staphylococcus aureus and Escherichia coli, which was suppressed by poly(I) and an anti-LOX-1 mAb . Adhesion of these bacteria to LOX-1 does not require divalent cations or serum factors and can be supported under both static and nonstatic conditions . Cultured bovine aortic endothelial cells (BAEC) can also support adhesion of FITC-labeled S . aureus, which was similarly suppressed by poly(I) and an anti-LOX-1 mAb . In contrast, binding of FITC-labeled E . coli to BAEC was partially inhibited by the anti-LOX-1 mAb, and poly(I) did not block FITC-labeled E . coli adhesion to BAEC, but, rather, enhanced it under a static condition . TNF-alpha increased LOX-1-dependent adhesion of E . coli, but not that of S . aureus, suggesting that S . aureus adhesion to BAEC may require additional molecules, which cooperate with LOX-1 and suppressed by TNF-alpha . Taken together, LOX-1 can work as a cell surface receptor for Gram-positive and Gram-negative bacteria, such as S . aureus and E . coli, in a mechanism similar to that of class A scavenger receptors; however, other unknown molecules may also be involved in the adhesion of E . coli to BAEC, which is enhanced by poly(I). J Immunol, 2001 Apr 15, 166(8), 4884 - 90 Unique functions of CD11b+, CD8 alpha+, and double-negative Peyer's patch dendritic cells; Iwasaki A et al.; We have recently demonstrated the presence of three populations of dendritic cells (DC) in the murine Peyer's patch . CD11b(+)/CD8alpha(-) (myeloid) DCs are localized in the subepithelial dome, CD11b(-)/CD8alpha(+) (lymphoid) DCs in the interfollicular regions, and CD11b(-)/CD8alpha(-) (double-negative; DN) DCs at both sites . We now describe the presence of a novel population of intraepithelial DN DCs within the follicle-associated epithelium and demonstrate a predominance of DN DCs only in mucosal lymphoid tissues . Furthermore, we demonstrate that all DC subpopulations maintain their surface phenotype upon maturation in vitro, and secrete a distinct pattern of cytokines upon exposure to T cell and microbial stimuli . Only myeloid DCs from the PP produce high levels of IL-10 upon stimulation with soluble CD40 ligand(-) trimer, or Staphylococcus aureus and IFN-gamma . In contrast, lymphoid and DN, but not myeloid DCs, produce IL-12p70 following microbial stimulation, whereas no DC subset produces IL-12p70 in response to CD40 ligand trimer . Finally, we show that myeloid DCs from the PP are particularly capable of priming naive T cells to secrete high levels of IL-4 and IL-10, when compared with those from nonmucosal sites, while lymphoid and DN DCs from all tissues prime for IFN-gamma production . These findings thus suggest that DC subsets within mucosal tissues have unique immune inductive capacities. J Hosp Infect, 2001 Apr, 47(4), 294 - 300 Emergence and spread of low-level mupirocin resistance in methicillin-resistant Staphylococcus aureus isolated from a community hospital in Japan; Watanabe H et al.; The objective of this study was to investigate the state of mupirocin resistance in methicillin-resistant Staphylococcus aureus (MRSA) in a community hospital in Japan . Ninety strains of MRSA were isolated from the respiratory tract of 56 patients (group I, Jun 1990-Aug 1996) before introduction of mupirocin in Japan, which were compared with 168 strains from 48 patients (group II, Sept 1996-Jan 1998) and 146 strains from 85 patients (group III, Feb 1999-Dec 1999) isolated after introduction of mupirocin . Comparisons were made by determining the minimum inhibitory concentrations (MIC) against nine antibiotics . Fifty-five MRSA isolates from 27 patients {13 (27.1%) of 48 in group II and 14 (16.5%) of 85 in group III} after introduction of mupirocin showed low-level resistance to mupirocin (MIC, 6.25 to 50 microg/ml) but the remaining isolates were sensitive to mupirocin (MIC < or =3.13 microg/ml) . Most patients colonized with low-level mupirocin-resistant MRSA were elderly (> or =65 years of age), on total parenteral nutrition or nasal feeding and had other underlying diseases . The proportion of patients colonized with low-level mupirocin-resistant MRSA following repeated use of mupirocin was higher in patients of group II than those of group III . Molecular typing by pulsed-field gel electrophoresis (PFGE) demonstrated that the pattern of 13 MRSA isolates from 13 patients of group II consisted of three patterns (A, B, C) with predominance of pattern A, while the pattern of 13 MRSA isolates from 13 patients of group III consisted of three patterns (A, C, D) with predominance of patterns A and D . Our results indicated that resistance of MRSA to mupirocin remains at a low level at present in Japan . However, we should be aware of the possible emergence of MRSA highly resistant to mupirocin in the future . J Korean Med Sci, 2001 Feb, 16(1), 39 - 44 Enhanced neutrophil functions by recombinant human granulocyte colony-stimulating factor in diabetic patients with foot infections in vitro; Peck KR et al.; This study was performed to evaluate the effect of granulocyte-colony stimulating factor on neutrophil functions in diabetic patients with active foot infections in vitro . Twelve diabetic patients with foot infections and 12 normal volunteers were enrolled . Neutrophils from peripheral blood were incubated with granulocyte colony-stimulating factor (G-CSF, 50 ng/mL) for 20 min . Superoxide production of neutrophils was measured by the reduction of ferricytochrome C . Neutrophil phagocytosis was assayed using Staphylococcus aureus and the weighted phagocytic index was calculated . Superoxide production of neutrophils in diabetic patients with foot infections was 7.7 (unit: nmol/2 x 10(5) cells/60 min), which was significantly lower than that in controls (12.0) (p<0.05) . G-CSF increased neutrophil superoxide production to 12.1 in diabetic patients with foot infections and to 19.8 in controls (p<0.05 for each) . Weighted phagocytic index in diabetic patients with foot infections was 0.77, which was not significantly different from that of the controls (0.69) . Weighted phagocytic index was increased significantly by G-CSF to 0.88 in diabetic patients with foot infections and to 0.79 in controls (p<0.05 for each) . In conclusion, G-CSF significantly enhanced neutrophil functions in diabetic patients with foot infections in vitro. J Nat Toxins, 2001 Feb, 10(1), 1 - 8 Genes encoding staphylococcal enterotoxins G and I are linked and separated by DNA related to other staphylococcal enterotoxins; Monday SR et al.; Fifteen randomly selected Staphylococcus aureus isolates, known to carry the staphylococcal enterotoxin (SE) G determinant (seg), were shown to carry the SEI determinant (sei) . To determine whether these two genes are linked, two S . aureus strains (FRI445 and FRI572), each containing the seg and sei determinants, were further analyzed . In these strains, sei is located 2,002 bp upstream of seg . Within the intergenic nucleotide sequence are three regions of nucleotide sequence with significant identity to the sequences of other SE genes . Characterization of the DNA regions surrounding the seg and sei determinants will allow a better understanding of the association between these two genes and may explain why they are so frequently observed simultaneously in S . aureus isolates. Med Dosw Mikrobiol, 2000, 52(4), 397 - 403 {The effect of selected factors on the course of wound healing after surgery for laryngeal neoplasms}; Jozefowicz-Korczynska M et al.; Finding of the a etiologic factors and participation of bacteria flora in wound healing in laryngeal cancer treatment was the purpose of our study . Investigations were performed in 27 patients . Swabs were taken from the postoperative wounds . Detailed identifications of the bacteria flora and antibiotic susceptibility of bacteria were performed . Wound healing was estimated according to extension of the carcinoma, applied antibiotics, state of the oral cavity, the kind of bacteriological flora isolated after surgical treatment from postoperative wounds . It was found that wound healing depended on the extension of carcinoma, as well as, type of isolated bacteria and antibiotic therapy used . The proper healing of postoperative wounds was not dependent on the state of the oral cavity and the dentition . The main cause of postoperative complication of wounds was methicillin-resistant Staphylococcus aureus (MRSA). Med Dosw Mikrobiol, 2000, 52(4), 327 - 32 {Evaluation of methicillin-resistance in Staphylococcus aureus by the agar disk diffusion method and PCR}; Idzik D et al.; Staphylococcus aureus is a widespread human pathogen . One the most striking characteritics of this bacterium is resistance to methicillin and all beta-lactam antibiotics . The agar disk diffusion method is the most widely used in vitro susceptibility test, but recently molecular methods, e.g . Polymerase Chain Reaction, have been also introduced . We compared the detection of methicillin resistant coagulase positive Staphylococcus aureus isolated from clinical materials in Silesian microbiological laboratories by diffusion method and PCR through the detection of nuc and mec A genes . Our results show that PCR used for the detection of mec A gene increases the detection of methicillin-resistant Staphylococcus aureus strains by 10% as compared to the agar disk diffusion method . Among Staphylococcus aureus strains, detected as methicillin-resistant, 17% of organisms showed no presence of mec A gene. Biochemistry, 2001 Apr 10, 40(14), 4426 - 36 Spectroscopic properties of the metalloregulatory Cd(II) and Pb(II) sites of S . aureus pI258 CadC; Busenlehner LS et al.; Staphylococcus aureus pI258 CadC is an extrachromosomally encoded metalloregulatory repressor protein from the ArsR superfamily which negatively regulates the expression of the cad operon in a metal-dependent fashion . The metalloregulatory hypothesis holds that direct binding of thiophilic divalent cations including Cd(II), Pb(II), and Zn(II) by CadC allosterically regulates the DNA binding activity of CadC to the cad operator/promoter (O/P) . This report presents a detailed characterization of the metal binding and DNA binding properties of wild-type CadC . The results of analytical ultracentrifugation experiments suggest that both apo- and Cd(1)-CadC are stable or weakly dissociable homodimers characterized by a K(dimer) = 3.0 x 10(6) M(-1) (pH 7.0, 0.20 M NaCl, 25.0 degrees C) with little detectable effect of Cd(II) on the dimerization equilibrium . As determined by optical spectroscopy, the stoichiometry of Cd(II) and Pb(II) binding is approximately 0.7-0.8 mol/mol of wild-type CadC monomer . Chelator (EDTA) competition binding isotherms reveal that Cd(II) binds very tightly, with K(Cd) = 4.3 (+/-1.8) x 10(12) M(-1) . The results of UV-Vis and X-ray absorption spectroscopy of the Cd(1) complex are consistent with a tetrathiolate (S(4)) complex formed by four cysteine ligands . The (113)Cd NMR spectrum reveals a single resonance of delta = 622 ppm, consistent with an S(3)(N,O) or unusual upfield-shifted S(4) complex . The Pb(II) complex reveals two prominent absorption bands at 350 nm (epsilon = 4000 M(-1) cm(-1)) and 250 nm (epsilon = 41 000 M(-1) cm(-1)), spectral properties consistent with three or four thiolate ligands to the Pb(II) ion . The change in the anisotropy of a fluorescein-labeled oligonucleotide containing the cad O/P upon binding CadC and analyzed using a dissociable CadC dimer binding model reveals that apo-CadC forms a high-affinity complex {K(a) = (1.1 +/- 0.3) x 10(9) M(-1); pH 7.0, 0.40 M NaCl, 25 degrees C}, the affinity of which is reduced approximately 300-fold upon the binding of a single molar equivalent of Cd(II) or Pb(II) . The implications of these findings on the mechanism of metalloregulation are discussed. Biochemistry, 2001 Apr 10, 40(14), 4312 - 22 Analysis of MDR1 P-glycoprotein conformational changes in permeabilized cells using differential immunoreactivity; Druley TE et al.; The reactivity of the ATP-dependent multidrug transporter P-glycoprotein (Pgp) with the conformation-sensitive monoclonal antibody UIC2 is increased in the presence of Pgp transport substrates, ATP-depleting agents, or mutations that reduce the level of nucleotide binding by Pgp . We have investigated the effects of nucleotides and vinblastine, a Pgp transport substrate, on the UIC2 reactivity of Pgp in cells permeabilized by Staphylococcus aureus alpha-toxin . ATP, ADP, and nonhydrolyzable ATP analogues decreased the UIC2 reactivity; this effect was potentiated by vanadate, a nucleotide-trapping agent . The Hill number for the nucleotide-induced conformational transition was 2 for ATP and ADP but 1 for nonhydrolyzable ATP analogues . The Hill numbers for ATP and ADP were decreased to 1 by mutations in one of the two nucleotide binding sites of Pgp, whereas mutation of both sites greatly diminished the overall effect of nucleotides . Vinblastine reversed the decrease in the UIC2 reactivity brought about by all the nucleotides, including nonhydrolyzable analogues; this effect of vinblastine was blocked by vanadate . These data indicate that UIC2-detectable conformational changes of Pgp are driven by binding and debinding of nucleotides, that nucleotide hydrolysis affects the Hill number for its Pgp interactions, and that Pgp transport substrates promote nucleotide dissociation from Pgp . These findings are consistent with a conventional E1/E2 model that explains conformational transitions of a transporter protein through a series of linked equilibria. Microbiology, 2001 Apr, 147(Pt 4), 803 - 10 The signal transducer (BlaRI) and the repressor (BlaI) of the Staphylococcus aureus beta-lactamase operon are inducible; Clarke SR et al.; The precise start points for transcription of the blaZ and of the blaRI/blaI genes of the Staphylococcus aureus beta-lactamase operon have been determined by primer extension analysis . Consequently the overlapping promoter sequences were deduced . Northern blots showed that the synthesis of the 2100 nt mRNA from blaRI is inducible and that a blaI probe hybridized to the same mRNA as the blaRI probe . The gene cat, encoding chloramphenicol acetyltransferase, was fused separately to the blaZ and blaRI/blaI promoters, and used to compare their strengths . The promoter for blaZ is about six times stronger than that for blaRI/blaI and the synthesis of chloramphenicol acetyltransferase from both promoters is inducible, supporting the results from the Northern blots. J Clin Microbiol, 2001 Apr, 39(4), 1669 - 71 Low Prevalence of Community-Acquired Methicillin-Resistant Staphylococcus aureus in Adults at a University Hospital in the Central United States; Suntharam N et al.; Community-acquired MRSA (CA-MRSA) is potentially a new emerging pathogen with most strains susceptible to many antimicrobials except for beta-lactam antibiotics . We retrospectively reviewed MRSA isolates during a 20-month study period (January 1998 through August 1999) and investigated those that were clindamycin susceptible . Patients were not considered to harbor CA-MRSA if they had been admitted to a hospital within the preceding 2 years or if their isolate had been obtained more than 72 h after admission . There were 2,817 S . aureus isolates, with 1,071 (38%) being MRSA . Of these 1,071 isolates, 161 were clindamycin susceptible; these were recovered from 81 patients . Of these 81 patients, 20 appeared to have community-acquired strains, but only 2 could be confirmed as having CA-MRSA. J Clin Microbiol, 2001 Apr, 39(4), 1540 - 8 Identification and characterization of phage variants of a strain of epidemic methicillin-resistant Staphylococcus aureus (EMRSA-15); O'Neill GL et al.; EMRSA-15 is one of the most important strains of epidemic methicillin-resistant Staphylococcus aureus (EMRSA) found in the United Kingdom . It was originally characterized by weak lysis with phage 75 and production of enterotoxin C but not urease . Two variant strains of EMRSA-15 which show a broader phage pattern than the progenitor strain have emerged . A total of 153 recent clinical isolates representing classical EMRSA-15 (55 isolates) or these phage variants (98 isolates) were compared by SmaI macrorestriction profiles in pulsed-field gel electrophoresis (PFGE) as well as by urease and enterotoxin C production . Eight of the 98 isolates were shown to be other unrelated strains by both PFGE and their production of urease, a misidentification rate of 8% by phage typing . Seventy-one EMRSA-15 isolates were enterotoxin C negative, and the majority of these were sensitive to phage 81 . Examination of PFGE profiles and Southern blotting studies suggest that the enterotoxin C gene locus is encoded on a potentially mobile DNA segment of ca . 15 kb . After elimination of the eight non-EMRSA-15 isolates, the remaining 145 were characterized by PFGE, yielding 22 profiles . All profiles were within five band differences of at least one other profile . Classical EMRSA-15 isolates showed nine PFGE profiles, with the majority of isolates (68%) in profile B1 . Six of these nine PFGE profiles were unique to the classical EMRSA-15 isolates . Among the phage variants of EMRSA-15, 16 profiles were seen, but the majority of isolates (83%) fell into 1 of 4 profiles (B2, B3, B4, and B7) which correlated well with phage patterns . The most divergent PFGE profiles among the EMRSA-15 isolates had as many as 12 band differences from one another, suggesting that in examining isolates belonging to such a temporally and geographically disseminated epidemic strain, the range of PFGE profiles must be regarded as a continuum and analyzed by relating the profiles back to the most common or progenitor profile. Int J Pharm, 2001 Feb 19, 214(1-2), 93 - 8 Activity of mammalian secreted phospholipase A(2) from inflammatory peritoneal fluid towards PEG-liposomes . Early indications; Vermehren C et al.; Due to an increase in the activity of phospholipase A(2) (PLA(2)) in various inflammatory diseases, this enzyme may play a key role in the degradation of liposomes and the subsequent release of drug when PEG-liposomes passively target inflammatory tissue . The activity of mammalian secreted phospholipase A(2) (sPLA(2)) in casein stimulated peritoneal fluid was tested toward liposomes of different compositions . Early results indicate only a slight degradation of conventional dipalmitoylphosphatidylcholine (DPPC) liposomes as well as DPPC liposomes incorporated with different concentrations of PEG(2000) . However, the DPPC degradation increased to 7% when inclusion of 30 mol% phosphatidylethanolamine (PE) in the lipid bilayer . The increase in degradation may be due to an improvement of the substrate - as it is well known, that PE is a better substrate for the mammalian sPLA(2) than PC . Incorporation of PE into the bilayer may increase the binding properties of the bilayer resulting in improved conditions for the enzymatic attack by sPLA(2) . In addition, inhibitory zones of Staphylococcus aureus in an agar diffusion test showed that PLA(2) from Crotalus atrox venom was able to catalyze the release of gentamicin from PEG-liposomes . In conclusion, this study suggest that degradation of the lipid bilayer of PEG-liposomes by PLA(2) result in release of incapsulated drug, e.g . gentamicin and inclusion of PE in the liposomal bilayer, may enhance the activity of the mammalian sPLA(2) toward liposomes composed of DPPC. J Med Assoc Thai, 2001 Jan, 84(1), 63 - 8 Clinical features of septic arthritis of sternoclavicular joint; Akkasilpa S et al.; We studied 21 patients with septic arthritis of the sternoclavicular joint at Chulalongkorn University Hospital between January 1987 and January 1997 . There were 15 males (71.4%) and 6 females (28.6%) . The mean age was 47.4 years with a range of 16 to 69 . More than half of the patients (57.1%) were aged more than 50 years and most had associated diseases including diabetes mellitus and cirrhosis . Almost all of the younger age group had a history of intravenous drug abuse . All of the patients had fever and sternoclavicular joint pain . Most of the patients (66.7%) had monoarticular arthritis, whereas, the others had oligoarticular arthritis . Staphylococcus aureus was the most commonly or identified organism in the patients . Retrosternal abscess was seen by computerized tomography in 6 patients (28.6%) . All patients received parenteral antibiotics, and 5 patients (23.8%) required surgical drainage of a retrosternal abscess . Eighteen patients recovered but there were 3 (14.3%) deaths . All of these had retrosternal abscesses . The major cause of death was septic shock . Septic arthritis of the sternoclavicular joint is an uncommon disease in Thai clinical practice . Although uncommon, retrosternal abscess is a life threatening complication. Nihon Rinsho Meneki Gakkai Kaishi, 2001 Feb, 24(1), 48 - 56 {A boy diagnosed SLE after Staphylococcus aureus infection over a long period}; Wada Y et al.; We encountered a 13-year-old boy with SLE who showed specific pathophysiology . The affected child was hospitalized because of long-standing cutaneous empyesis considered to be Staphylococcus aureus infection, followed by manifestation of meningoencephalitis-like symptoms . On a close check up, the patient was diagnosed as having SLE complicated with interstitial lupus nephritis and verrucosis of the left ventricle . Besides the findings, the blastogenesis of the patient's lymphocyte was low against stimulation of sac-1 which connects with the Fc portion of lgG, one of the constituent proteins of Staphylococcus . Moreover, anti-phospholipid antibodies turned positive during immunosuppressive therapy and subcutaneous abscess due to Pseudomonas aeruginosae developed concurrently at about the same time, which posed difficulties in the treatment . The affected child had had Staphylococcus aureus infections over a long period of time before diagnosis of SLE and was susceptible to bacterial infections due to Pseudomonas aeruginosae during the treatment . The clinical course of this case was considered important in presuming the complex immunologically abnormal condition of SLE in childhood. Arch Intern Med, 2001 Feb 12, 161(3), 406 - 10 Single-lumen subcutaneous ports inserted by interventional radiologists in patients undergoing chemotherapy: incidence of infection and outcome of attempted catheter salvage; Kuizon D et al.; BACKGROUND: Subcutaneous ports are commonly used for vascular access in patients with cancer undergoing chemotherapy . OBJECTIVES: To determine the incidence of catheter-related infection and to assess the efficacy of catheter salvage in subcutaneous ports . METHODS: We retrospectively reviewed 300 subcutaneous single-lumen chest ports inserted by interventional radiologists in 294 patients between December 1, 1995, and November 15, 1997, at the Cleveland Clinic Foundation, Cleveland, Ohio . The number of days that the catheter remained in situ, infection rate, treatment, and outcome of infection were determined . RESULTS: Two hundred ninety-four patients had a total of 79 748 catheter-days . Vascular access for chemotherapy was the indication for 95% of the subcutaneous ports placed . Seventeen catheters (5.7%) developed 20 episodes of noninfectious complications resulting in the removal of 6 ports . Seventeen patients (5.7%) developed catheter-related infections (2.1/10 000 catheter-days) including 10 episodes of catheter-related bacteremia (1.2/10 000 catheter-days) . The most common organism isolated was Staphylococcus aureus . A total of 15 of the 17 infected catheters were removed . Salvage was attempted in 6 patients in whom 4 catheters were eventually removed due to recurrent bacteremia (2 patients) and persistent local infection (2 patients) . One of the 10 patients with catheter-related bacteremia developed septic arthritis . There were no complications associated with attempted catheter salvage . CONCLUSIONS: Subcutaneous single-lumen ports inserted by interventional radiologists in patients undergoing chemotherapy have low complication rates but infections remain the leading cause of catheter loss . Antibiotic therapy without catheter removal is unlikely to eradicate catheter-related bacteremia. Commun Dis Public Health, 2000 Dec, 3(4), 288 - 90 Infection of foot ulcers with Staphylococcus aureus associated with increased mortality in diabetic patients; Mantey I et al.; Diabetic patients with foot ulceration have a poorer prognosis than those without ulceration . The reason for this is unclear, but there is considerable interest in the putative links between infection and atherogenesis, and it is notable that diabetic foot ulcers (DFU) are often infected with Staphylococcus aureus and the main cause of death in DFU patients is ischaemic heart disease . We examined the 5 year survival of 71 diabetic patients who presented with foot ulcers that were newly infected (Sa group, n = 56) or not infected at all during the study period (non-Sa group, n = 15) with S . aureus . Twenty-nine patients (52%) infected with S . aureus died compared with three patients (20%) whose foot ulcers were not infected with S . aureus . The patients in the two groups were similar in age and duration of diabetes . The overall five year mortality rate was 10.4% per year for those infected, significantly higher than the average of 4.0% for patients without infection (p = 0.015) . None of the patients was bacteraemic or died directly from sepsis . Infection of DFU by S . aureus may increase the risk of death in diabetic patients. Indian J Med Res, 2001 Jan, 113, 11 - 3 Inactivation of chloramphenicol by Staphylococcus aureus biotype C from humans & animals; Dutta GN et al.; During an investigation, 55 biotype C (bovine and caprine biotype) Staphylococcus aureus isolated from 43 cows suffering from mastitis, and 20 biotype C Staph . aureus strains from the nares and the side of nail-tips of the right thumbs of 20 farm workers (milkers and animal attendants) on six small dairy farms in Assam and Meghalaya were isolated . Three strains from the former and six strains from the latter from among the isolates on two of the farms were found resistant to chloramphenicol, when tested with a routine susceptibility test . Test of the organisms by the agar dilution method indicated that the resistant strains had a minimal inhibitory concentration for chloramphenicol of 32 micrograms/ml or more, while, the chloramphenicol sensitive strains and two reference strains, Staph . aureus ATCC 25923 and Micrococcus luteus ATCC 9341, had < or = 8 micrograms/ml . Two bovine and five human chloramphenicol resistant strains showed positive results when tested by the Gots test . When these strains were tested by a standard method in broth, containing 30 micrograms chloramphenicol per ml, all showed evidence of inactivating chloramphenicol up to a non-detectable level within 36 h . Inactivation of chloramphenicol by Staph . aureus has clinical significance. J UOEH, 2001 Mar 1, 23(1), 59 - 67 {A case of pemphigus foliaceus associated with bullous impetigo successfully treated with tetracycline and nicotinamide}; Izu K et al.; A 50-year-old Japanese woman visited our clinic, complaining of generalized erythema with painful erosions and bullae . The histopathological findings of the skin lesion suggested development of impetigo . Gentamycin-resistant Staphylococcus aureus was detected by the bacterial culture examination from the impetiginous bullae . A direct immunofluorescence study of the lesion showed an intercellular deposition of IgG and C3 in the upper epidermis . We diagnosed this case as pemphigus foliaceus associated with bullous impetigo . A combined oral administration of tetracycline (200 mg/day) and nicotinamide (1200 mg/day) for 3 weeks was successful . In Japan, patients with moderate to severe symptoms of pemphigus foliaceus are usually treated with oral steroid therapy . To our knowledge, however, there is no reported pemphigus case which has been successfully treated only with tetracycline and nicotinamide. J Biol Chem, 2001 May 4, 276(18), 14955 - 60 Epub 2001 Feb 14. Role of cysteinyl residues in sensing Pb(II), Cd(II), and Zn(II) by the plasmid pI258 CadC repressor; Sun Y et al.; The cadCA operon of Staphylococcus aureus plasmid pI258 confers resistance to salts of the soft metals lead, cadmium, and zinc . The operon is regulated by CadC, a member of the ArsR family of metal-responsive transcriptional repressors . In this study the role of the five cysteine residues of CadC in soft metal ion sensing was investigated . Cys-7, Cys-11, Cys-52, Cys-58, and Cys-60 were changed individually to glycine or serine residues . The effect of the cadC mutations was examined in Escherichia coli using a green fluorescent protein reporter system . None of the mutations affected the ability of CadC to repress gfp expression . Neither Cys-11 nor Cys-52 was required for in vivo response to Pb(II), Zn(II), or Cd(II) . Cys-7, Cys-58, or Cys-60 mutations each reduced or eliminated soft metal sensing . Wild-type and mutant CadC proteins were purified, and the effect of the substitutions on DNA binding was determined using a restriction enzyme protection assay . Binding of wild-type CadC protected cad operator DNA from digestion at the single SspI site, and the addition of Pb(II), Zn(II), or Cd(II) resulted in deprotection . Chemical modification of the cysteine residues in CadC had no effect on protection but eliminated deprotection . C11G and C52G proteins exhibited wild-type properties in vitro . C7G, C58S, and C60G proteins were able to be protected from SspI digestion but had reduced responses to soft metal ions . The results indicate that Cys-7, Cys-58, and Cys-60 are involved in sensing those soft metals and suggest that they are ligands to Pb(II), Zn(II), and Cd(II). J Nat Prod, 2001 Mar, 64(3), 399 - 400 Daucane sesquiterpenes from Ferula hermonis; Galal AM et al.; The roots of Ferula hermonis Boiss yielded two new daucane esters, 14-(4'-hydroxybenzoyloxy)dauc-4,8-diene (1) and 14-(4'-hydroxy-3'-methoxybenzoyloxy)dauc-4,8-diene (2), together with the four known sesquiterpenes jaeschkeanadiol p-hydroxybenzoate (3), jaeschkeanadiol benzoate (4), jaeschkeanadiol (5), and epoxyjaeschkeanadiol (6) . The identities of the isolated compounds were ascertained primarily using NMR and MS data . Compounds 1 and 3 exhibited antimicrobial activity against Staphylococcus aureus with IC(50) 1.5 and 3.5 microg/mL, respectively, and against Methicillin-resistant S . aureus with IC(50) 2.0 and 4.0 microg/mL, respectively. Bioorg Med Chem Lett, 2001 Mar 26, 11(6), 797 - 801 Anti-MRSA cephems . Part 1: C-3 substituted thiopyridinium derivatives; Springer DM et al.; Sixteen novel cephalosporin derivatives with activity against methicillin-resistant Staphylococcus aureus (MRSA) are described . The compounds were synthesized using substituted thiopyridones, generated either by cyclization of functionalized precursors, or by direct alkylation of the enolate of 2-methyl substituted pyrones . The most active compound in vitro against a strain of MRSA (A27223) displayed an MIC of 0.5 microg/mL . The most efficacious compound in vivo had a PD50 of 2.1 mg/kg. J Pept Sci, 2001 Feb, 7(2), 74 - 81 Antibiotic activity of pentadecapeptides modelled from amino acid descriptors; Lejon T et al.; Pentadecapeptides based on modified murine lactoferricin (LFM) sequences show varying degrees of antibacterial activity against Escherichia coli and Staphylococcus aureus . By means of projections to latent structures (PLS), a good correlation is obtained if the biological activity is modelled as a function of variables describing peptide properties, e.g . alpha-helicity, hydrophobicity/hydrophilicity and charge . Using variables derived from a principal component analysis (PCA) of all naturally occurring amino acids, it is possible to describe the amino acid content of the peptides using three variables per amino acid position . The resulting descriptor matrix is then used to develop quantitative structure-activity relationships (QSAR) . It is shown that the theoretically derived descriptors model the activity of the peptides better than the earlier model, and that properties of the peptides other than antibacterial activity can be predicted. J Bacteriol, 2001 Apr, 183(8), 2417 - 24 Recruitment of the mecA gene homologue of Staphylococcus sciuri into a resistance determinant and expression of the resistant phenotype in Staphylococcus aureus; Wu SW et al.; Strains of methicillin-resistant Staphylococcus aureus (MRSA) have become the most important causative agents of hospital-acquired diseases worldwide . The genetic determinant of resistance, mecA, is not a gene native to S . aureus but was acquired from an extraspecies source by an unknown mechanism . We recently identified a close homologue of this gene in isolates of Staphylococcus sciuri, a taxonomically primitive staphylococcal species recovered most frequently from rodents and primitive mammals . In spite of the close sequence similarity between the mecA homologue of S . sciuri and the antibiotic resistance determinant mecA of S . aureus, S . sciuri strains were found to be uniformly susceptible to beta-lactam antibiotics . In an attempt to activate the apparently "silent" mecA gene of S . sciuri, a methicillin-resistant derivative, K1M200 (for which the MIC of methicillin is 200 microg/ml), was obtained through stepwise exposure of the parental strain S . sciuri K1 (methicillin MIC of 4 microg/ml) to increasing concentrations of methicillin . DNA sequencing of the mecA homologue from K1M200 revealed the introduction of a point mutation into the -10 consensus of the promoter: the replacement of a thymine residue at nucleotide 1577 in the susceptible strain K1 by adenine in the resistant strain K1M200, which was accompanied by a drastic increase in transcription rate and the appearance of a new protein that reacted with monoclonal antibody prepared against the penicillin-binding protein 2A (PBP2A), i.e., the gene product of S . aureus mecA . Transduction of mecA from K1M200 (cloned into a plasmid vector) into a methicillin-susceptible S . aureus mutant resulted in a significant increase of methicillin resistance (from a methicillin MIC of 4 micro/ml to 12 and up to 50 microg/ml), the appearance of a low-affinity PBP detectable by the fluorographic assay, and the production of a protein that reacted in a Western blot with monoclonal antibody to PBP2A . Antibiotic resistance and the protein products disappeared upon removal of the plasmid-borne mecA homologue . The observations support the proposition that the mecA homologue ubiquitous in the antibiotic-susceptible animal species S . sciuri may be an evolutionary precursor of the methicillin resistance gene mecA of the pathogenic strains of MRSA. Am J Kidney Dis, 2001 Apr, 37(4), 815 - 9 Heat-killed microorganisms induce PAI-1 expression in human peritoneal mesothelial cells: role of interleukin-1alpha; Mandl-Weber S et al.; Human peritoneal mesothelial cells (HMCs) have a critical role in maintaining the intraperitoneal balance between fibrinolysis and coagulation by expressing the fibrinolytic enzyme, tissue-type plasminogen activator (tPA), as well as a specific plasminogen activator inhibitor (type 1; PAI-1) . During bacterial peritonitis, the balance between intraperitoneal generation and degradation of fibrin is disturbed . As a consequence, severe peritoneal damage occurs, which is one of the leading causes of patient dropout from continuous ambulatory peritoneal dialysis (CAPD) therapy . Cultured HMCs isolated from omental biopsy specimens were used to study the effect of heat-killed strains (2 x 10(8)/mL) of Staphylococcus aureus, Staphylococcus epidermidis, and Escherichia coli on the synthesis of tPA and PAI-1 . Conditioned media were obtained by incubating cells with the different bacterial strains . tPA and PAI-1 antigen concentrations were measured in the cell supernatants by enzyme-linked immunosorbent assay . Each of the three heat-killed microorganisms induced a time-dependent increase in PAI-1 synthesis . After a 48-hour incubation period, the strongest effect was seen in the presence of S aureus (3.5-fold versus control), followed by S epidermidis (2.5-fold versus control) and E coli (1.5-fold versus control) . Under the same conditions, tPA antigen levels did not change after exposure to S aureus or E coli, whereas the addition of S epidermidis resulted in enhanced tPA antigen production (2-fold versus control) . The increase in PAI-1 synthesis in the presence of the heat-killed microorganisms was preceded by similar changes in interleukin-1alpha (IL-1alpha) levels . Inhibiting the activity of IL-1alpha with a neutralizing antibody significantly reduced bacterial-induced PAI-1 production . Our results indicate that the fibrinolytic imbalance during bacterial peritonitis depends on the bacterial species . The increase in PAI-1 synthesis, not the decrease in the production of tPA, alters mesothelial fibrinolytic activity . Because the increase in PAI-1 expression is significantly quenched by blocking the activity of IL-1alpha, the mesothelial release of this cytokine is involved in bacterial-induced changes in the fibrinolytic system. Am J Med Sci, 2001 Feb, 321(2), 152 - 5 The role of transesophageal echocardiography in the diagnosis and management of patients with aortic perivalvular abscesses; Lerakis S et al.; Aortic valve abscesses (AVAs) are a devastating complication of aortic valve endocarditis . Over 8 years, 25 patients were diagnosed with AVA by transesophageal echo (TEE) . Management and outcomes were then analyzed . Eleven (44%) AVAs involved prosthetic valves, and 6 (24%) occurred in congenitally malformed valves . Twenty patients (80%) underwent surgical intervention; the rest were treated medically . Eleven (44%) of the patients died {6 (30%) surgery patients and all the medical patients} . Eight of 11 (73%) patients who died were culture positive for Staphylococcus aureus . All patients with congenitally malformed aortic valves underwent surgical intervention and survived . We conclude that: (1) despite advances in therapy and diagnosis, patients with AVAs have a high mortality rate; (2) prognosis with AVA is especially poor when S aureus is the infectious organism; (3) patients with AVAs in congenitally malformed valves have a great outcome with surgery; (4) patients treated medically have a very poor prognosis; earlier identification by TEE may be critical to improving survival. CRNA, 2000 Feb, 11(1), 8 - 14 Infectious diseases in the operating room; Homa DG et al.; Patients with infectious diseases have special implications for infection control in the operating room . The increased use and abuse of antibiotics has ushered in a category of resistant organisms . These multiresistant organisms are spread by direct or indirect contact, primarily from the hands of caregivers or contact with contaminated environmental surfaces . Another category of infectious diseases is prions (pronounced pree-ons) . Unlike other infectious diseases, human prions diseases are not spread through routine exposures such as direct contact, droplet, and airborne routes . The causative agent is highly resistant to traditional disinfecting and sterilization processes . This article provides an overview of the multiresistant infections of methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, and Staphylococcus aureus with reduced susceptibility to vancomycin along with the human prions diseases Creutzfeldt-Jakob disease, German-Straussler-Scheinker syndrome, kuru, and fatal familial insomnia . We provide a template of precautions that can be used in developing operating room and anesthesia infection control protocols for this patient population. Curr Microbiol, 2001 Mar, 42(3), 203 - 10 Structure-activity relationships for six ketolide antibiotics; Champney WS et al.; Six structurally related 3-keto-substituted macrolide antibiotics (ketolides) were compared for concentration-dependent inhibitory effects on growth rate, viable cell number, and protein synthesis rates in Staphylococcus aureus cells . Inhibitory effects on 50S ribosomal subunit formation were also examined, as this is a second target for these antibiotics . A concentration range of 0.01 to 0.1 microg/ml was tested . An IC50 for inhibition of translation and 50S synthesis was measured for each compound, to relate structural features to inhibitory activity . ABT-773 was the most effective of the six compounds tested with an IC50 = 0.035 microg/ml . HMR 3004 was almost as effective with an IC50 = 0.05 microg/ml . Two 2-fluoroketolides (HMR 3562 and HMR 3787) were equivalent in their inhibitory activity with an IC50 = 0.06 microg/ml . Telithromycin (HMR 3647) had an IC50 = 0.08 microg/ml, and HMR 3832 was least effective with an IC50 = 0.11 microg/ml . Each antibiotic had an equivalent inhibitory effect on translation and 50S subunit formation . These results indicate specific structural features of these antimicrobial agents, which contribute to defined inhibitory activities against susceptible organisms. Microbiol Immunol, 2001, 45(1), 23 - 7 Relatedness between the coagulase gene 3'-end region and coagulase serotypes among Staphylococcus aureus strains; Kanemitsu K et al.; The 3'-end region of the coagulase gene from 22 strains of Staphylococcus aureus including 10 standard serotype strains was sequenced, and five subgroups with 4-8 tandem repeating units were distinguished among the tested strains . Phylogenetic analysis of the 3'-end region of the coagulase gene indicated that strains belonging to the same serotype were clustered in the same branch . A phylogenetic tree of the deduced amino acid sequences revealed that the C-terminal region might not be responsible for the epitope of the coagulase protein. Cutis, 2001 Mar, 67(3), 243 - 5 Tufted hair folliculitis after scalp injury; Fernandes JC et al.; We describe the case of a 38-year-old epileptic man with tufted hair folliculitis . The condition started 5 years ago after a scalp laceration that had been sustained 3 months earlier during an epileptic crisis . There then appeared a circumscribed inflammatory bulging lesion (with exudation and crusts) that evolved to scarring alopecia with tufts of 20 to 30 apparently normal hair shafts . Results of bacteriologic examination of pus extruding from the dilated follicular ostia revealed Staphylococcus aureus . The cutaneous pathologic examination showed polymorphous inflammatory exudate in the upper and mid dermis, which was mostly perifollicular, and the presence of normal and independent follicles in the deep dermis, which, while ascending, converged to a common dilated follicular channel . The patient was treated successively with oral flucloxacillin, erythromycin, ciprofloxacin, and amoxicillin/clavulanic acid and with topical application of erythromycin, clindamycin, povidone iodine, and ketoconazole . Transient improvement was followed by recurrence and enlargement of the affected area. Allergol Immunopathol (Madr), 2000 Nov-Dec, 28(6), 328 - 31 Repeated furunculosis in adult male with abnormal neutrophil activity; Forte WC et al.; A 21 years old male suffered from repeated furunculosis in different regions of the body over the last two years . This coincided with the start of professional activities in hospital surroundings . The purulent secretions all showed growth of Staphylococcus aureus . All laboratory tests were normal except for a decrease of the neutrophil phagocytic ingestion phase . Before the diagnosis of defective phagocytosis was made, antibiotic treatment was started about 4 to 5 days after the appearance of the infectious process and the furunculosis led to abscess formation with difficult healing and cellulitis . After the diagnosis of defective phagocytosis ingestion phase, personal hygiene was intensified during and after work shifts at the hospital and antibiotic treatment was started at the first signs of folliculitis, which showed healing. J Microbiol Immunol Infect, 2000 Dec, 33(4), 267 - 70 Neonatal fungemia caused by Hansenula anomala: a case report; Ma JS et al.; Hansenula anomala, an ascosporogenous yeast of the class Ascomycetes, is a free-living organism isolated from the environment . It is also a part of the normal or transient flora of the human throat and alimentary tract . It has been recognized as an opportunistic pathogen and its infection is very rare . A premature infant, a victim of right femoral osteomyelitis and right hip arthritis caused by oxacillin-resistant Staphylococcus aureus, was found to have developed H . anomala fungemia just before the initiation of the antimicrobial therapy with teicoplanin . Antifungal agents (fluconazole and amphotericin B) were prescribed for 10 days despite the absence of clinical sign of systemic fungal infection . His general condition remained good, with a subsequent sterile blood culture . The patient was discharged after completing 5 weeks of antimicrobial therapy, and he remained well during follow-up at our outpatient clinics . Here, we also review the risk factors, the clinical presentations, and the therapeutic strategies of H . anomala infection in the literature. Soud Lek, 2001 Jan, 46(1), 5 - 8 {Age determination using peptide mapping of non-collagenous proteins in human dentin}; Sajdok J et al.; The submitted paper deals with possibilities of assessment of human age based on findings of stereospecific breakdown of proteins containing D-forms of aspartic acid . The stereospecificity of enzymatic breakdown assumes that after enzymatic hydrolysis peptide breakdown products with different molecular weights--the so-called peptide map--will be formed, depending how many D-aspartyl residues the protein contains . The authors proved in the submitted preliminary study in subjects of different age the formation of breakdown products of different size in non-collagenous proteins of human dentin which was hydrolyzed by protease V8 from Staphylococcus aureus. Arch Intern Med, 2001 Mar 26, 161(6), 859 - 63 Eradication of methicillin-resistant Staphylococcus aureus from a health center ward and associated nursing home; Kotilainen P et al.; BACKGROUND: Long-term health care facilities have been recognized as reservoirs of multiresistant bacterial strains, especially methicillin-resistant Staphylococcus aureus (MRSA) . Efforts to control MRSA in this setting usually have been only partially effective . We describe herein the eradication of epidemic MRSA from a Finnish health care center ward and affiliated nursing home . METHODS: The methods to control MRSA included (1) contact isolation precautions, (2) screening for asymptomatic carriage, (3) eradication of carriage, and (4) education of staff on hygienic measures . The first 6 patients with MRSA-positive findings were referred without delay to the Infectious Diseases Unit of the adjacent university hospital for eradication treatment . Later, an isolation unit of 6 rooms was founded in the health care center, where the MRSA-colonized patients were nursed as a separate cohort until they, in succession, were referred to the Infectious Diseases Unit for decolonization . RESULTS: From May 20 through August 17, 1993, the epidemic MRSA strain was isolated from 8 long-term patients on the 40-bed ward of the health care center, 4 of the 59 residents of the nursing home, and 1 member of the staff . Eradication of carriage was successful in all except 1 patient with dementia, who was nursed in contact isolation in the health care center until his death 21 months later . CONCLUSIONS: It is possible to eradicate MRSA from a long-term health care facility even after 13 cases by applying strict control measures . Our experience may be valuable in the future decision-making process for control of new and more challenging multiresistant bacteria, eg, vancomycin-resistant strains of MRSA. Protein Sci, 2001 Feb, 10(2), 434 - 44 Coupling of antibodies via protein Z on modified polyoma virus-like particles; Gleiter S et al.; Therapeutic application of virus-based delivery systems often implies a change of the tropism of these vectors . This can be achieved by insertion of polypeptides (e.g., antibody fragments) in viral coat proteins . Such fusion proteins have only been used in viral vectors so far and, as part of a virus, they have not been available for a detailed biophysical characterization . We analyzed a fusion protein called VP1-Z, which is based on the polyoma virus coat protein VP1 and protein Z . Protein Z is an engineered antibody-binding domain derived from protein A from Staphylococcus aureus . The fusion VP1-Z was constructed by insertion of protein Z in the HI-loop of VP1 . As wild-type VP1, VP1-Z formed pentameric capsomers and assembled to VLPs in vitro . The stability of these particles was very similar compared to that of VLPs of wild-type VP1 . Protein Z was fully structured in the fusion protein and was still capable of binding antibodies on the surface of VLPs of VP1-Z . Using this fusion protein, we could change the tropism of polyoma VLPs toward cells presenting on their surface the antigen of the coupled antibody. J Antimicrob Chemother, 2001 Apr, 47(4), 399 - 403 A modified population analysis profile (PAP) method to detect hetero-resistance to vancomycin in Staphylococcus aureus in a UK hospital; Wootton M et al.; One hundred methicillin-resistant Staphylococcus aureus (MRSA) strains, isolated between 1983 and 1999, were tested alongside the vancomycin hetero-resistant S . aureus (hVRSA) strain Mu 3, and vancomycin-resistant S . aureus (VRSA) strain Mu 50, for their resistance to vancomycin . This was achieved using the screening method described by Hiramatsu, gradient plates, agar incorporation, standard Etest, macrodilution Etest and a modified population analysis . Using Hiramatsu's screening method, 5% of the 100 MRSA were identified as VRSA and 5% identified as hVRSA, the gradient plates identified 7% hVRSA, and the standard and macrodilution Etests identified no hVRSA . Mu 3 appeared to be vancomycin-susceptible using both the agar incorporation and standard Etest methods, but was classified as hVRSA using the macrodilution Etest . The modified population analysis reliably detected vancomycin hetero-resistance in Mu 3 and identified no hVRSAs within the 100 MRSA sample. J Antimicrob Chemother, 2001 Apr, 47(4), 377 - 89 Studies of the operator region of the Staphylococcus aureus beta-lactamase operon; Clarke SR et al.; The repressor proteins BlaI and MecI bind similarly to the bla operator implicated in the regulation of beta-lactamase synthesis in Staphylococcus aureus . BlaI binds to two separate dyads but neither copper-phenanthroline footprinting nor dimethyl sulphate (DMS) methylation protection assays produced any evidence of a change in the geometry of the DNA between the two dyads . It is concluded that BlaI molecules bound at the dyads probably do not cause bending or looping of the intervening DNA . DMS protection assays of BlaI binding to the bla operator in vitro and in vivo gave similar results so that it is tentatively concluded that the in vitro results are an accurate reflection of the in vivo situation . Deletion of the dyad nearest to the blaZ gene resulted in decreased synthesis of the chloramphenicol acetyltransferase reporter protein synthesized from the blaZ promoter/translation initiator . Explanations for this are considered. MMWR Morb Mortal Wkly Rep, 2000 Jan 7, 48(51-52), 1165 - 7 Staphylococcus aureus with reduced susceptibility to vancomycin--Illinois, 1999; Microbial colonization of soft contact lenses over time; Cornea and Contact Lens Research Unit, School of Optometry and Cooperative Research Centre for Eye Research and Technology, University of New South Wales, Sydney, AustraliaPURPOSE: To compare the bacterial colonization of soft contact lenses in subjects for successively increasing periods, up to 13 nights of wear . The aim of this study was to determine whether increasing the length of lens wear predisposed subjects to high levels of microbial colonization of lenses . METHODS: Subjects (N = 20) were divided into those with a prior history of adverse events (N = 6), gram-negative bacterial carriers (N = 6), and those with no previous history (N = 8) . RESULTS: There were no temporal changes in microbial colonization of lenses . Lenses from all wearers were colonized at least once during the study by gram-positive bacteria at low numbers (<10 cfu/ml) . Gram-negative bacteria colonized lenses at least once in 80% of all wearers . Lenses from gram-negative bacterial carriers were more frequently colonized by Staphylococcus aureus and Pseudomonas sp . compared with subjects with no previous history and subjects with a prior history of adverse events, respectively . Lenses from gram-negative bacterial carriers were less frequently colonized by a range of gram-positive bacteria compared with subjects with a prior history of adverse events . CONCLUSIONS: Increasing the length of lens wear up to 13 nights did not result in a predictable increase in bacterial colonization of contact lenses . Gram-positive bacteria were isolated frequently but in low numbers, whereas gram-negative bacteria were present sporadically. Eur J Immunol, 2001 Jan, 31(1), 243 - 9 IgG subclass switch capacity is low in switched and in IgM-only, but high in IgD+IgM+, post-germinal center (CD27+) human B cells; Werner-Favre C et al.; Recent studies have shown that in humans the germinal center reactions produce three types of V(D)J mutated B cells in similar proportions, i.e . Ig-switched, IgD-IgM+ (IgM-only) and IgD+IgM+ cells, and that together they form the CD27+ compartment of recirculating B cells . We investigated the Ig isotype switch capacity of these cells . Peripheral blood B subsets were sorted and IgG subclass secretion in presence or absence of IL-4 was compared in B cell assays which lead to Ig secretion in all (coculture with EL-4 thymoma cells) or only in CD27+ (CD40L stimulation) B cells . Already switched IgG+ B cells showed no significant sequential switch and IgM-only cells also had a low switch capacity, but IgD+CD27+ switched as much as IgD+CD27- B cells to all IgG subclasses . Thus, in switched B cells some alterations compromising further switch options occur frequently; IgM-only cells may result from aborted switch . However, IgD+CD27+ human B cells, extensively V(D)J mutated and "naive" regarding switch, build up a repertoire of B cells combining (1) novel cross-reactive specificities, (2) increased differentiation capacity (including after T-independent stimulation by Staphylococcus aureus Cowan I) and (3) the capacity to produce appropriate isotypes when they respond to novel pathogens. Pathol Biol (Paris), 2001 Feb, 49(1), 33 - 40 {Antibiotic resistance of Staphylococcus aureus in urban experience: 6 month study in Aquitaine}; Quentin C et al.; Antibiotic resistance of Staphylococcus aureus has been surveyed by eight city laboratories of the Aquitaine area, during a six month-period (january to june 1998) . Antibiotic susceptibility testing has been performed by the disk diffusion method, and the results have been collected and analysed using the SIRscan system . After elimination of the redundant strains, a total of 747 isolates has been retained . They were mainly isolated from pus (64%) collected from patients of the community (40%) or hospitalized in 30 private clinics or nursing homes . The percentages of resistant strains (community/institutions) were: benzylpenicillin: 90% (87/92%), oxacillin: 39% (23/50%), kanamycin: 37% (22/47%); gentamicin: 13% (8/16%), tobramycin: 37% (21/47%), amikacin: 21% (13/27%); netilmicin: 6% (5/7%), erythromycin: 33% (30/35%), spiramycin: 72% (77/69%), lincomycin: 24% (17/29%), pristinamycin: 2% (1/2%), tetracycline: 17% (14/19%); pefloxacin: 40% (25/50%), fosfomycin: 9% (6/12%), rifampicin: 10% (7/13%), fusidic acid: 14% (11/15%), cotrimoxazole and vancomycin: 0% . Meticillin-susceptible strains of S . aureus were mostly sensitive to other antibiotics (< or = 6% resistant strains, except for erythromycin: 22%) . Among meticillin-resistant S . aureus, heterogeneous strains with a KT phenotype, and various resistance patterns to the remaining antibiotics were predominant (61%), compared to the homogeneous strains with a KTG phenotype and multiresistant to the other antibiotics (32%) . The frequencies of resistant strains were highly variable depending on the specimen, the laboratory and the health care institution, except for cotrimoxazole and vancomycin which were always active. Pathol Biol (Paris), 2001 Feb, 49(1), 16 - 22 {Epidemic of Staphylococcus aureus nosocomial infections resistant to methicillin in a maternity ward}; Le Coq M et al.; Methicillin-resistant Staphylococcus aureus (MRSA) nosocomial infections frequently occur in the hospital environment, but their incidence is less often observed in neonates . In the present investigation, seventeen cases were recorded over a nine-week period (two cases per week) . Pulsed field gradient gel electrophoresis confirmed the clonal character of the strain . The hypothesis of manually-transmitted infection due to contamination from multiple sources was reinforced by the fact the epidemic persisted in spite of the elimination of the main human infectious source and an absence of risk factors determined by the case-control study . The role of environmental factors in the persistence of this outbreak of MRSA infection has been considered. J Bone Joint Surg Am, 2001 Mar, 83-A(3), 412 - 9 The efficacy of low-pressure lavage with different irrigating solutions to remove adherent bacteria from bone; Bhandari M et al.; BACKGROUND: Recent studies have suggested that high-pressure irrigation may have adverse effects on bone . However, the use of low-pressure irrigation may not remove all adherent bacteria from bone . The type of irrigating solution may be an important factor in the removal of adherent bacteria with pulsatile lavage . In this study, we compared the effects of various irrigating solutions on the number and function of osteoblasts and osteoclasts and we examined the effectiveness of these solutions in removing adherent bacteria from bone . METHODS: To examine the effect of irrigating solutions on the number and activity of osteoblasts, we isolated calvarial cells from newborn C57BI/6 mice and exposed the cells to equivalent concentrations of ethanol, povidoneiodine, liquid soap, antimicrobial wash (50 U/L of bacitracin), or chlorhexidine gluconate, for two, ten, or twenty minutes . The cells were then cultured in the presence of bone-nodule-enhancing medium (beta-glycerophosphate and ascorbic acid) for twenty-one days . The medium was changed every three or four days . Mineralized nodules were stained with alizarin red S, and osteoblasts were stained with a histochemical stain for alkaline phosphatase . Osteoclasts were identified with tartrate-resistant acid-phosphatase staining . In a second experiment, canine cortical tibiae were contaminated with Staphylococcus aureus for six hours and subjected to different irrigating solutions with or without low-pressure lavage . Bacterial colony-forming units were quantitated under each set of conditions . RESULTS: Each solution resulted in a time-dependent decrease in the number of calvarial osteoblasts and osteoclasts compared with that in the controls . The 1% soap solution resulted in greater preservation of both alkaline-phosphatase activity and bone-nodule formation than did the other solutions . Moreover, the soap solution preserved the number of osteoclasts to the greatest extent . The povidone-iodine and chlorhexidine-gluconate solutions resulted in the largest decline in bone-nodule formation, alkaline-phosphatase activity, and number of osteoclasts . Low-pressure pulsatile lavage with the soap solution removed the most bacteria from the contaminated tibia when compared with either the soap solution alone or low-pressure irrigation with saline solution . CONCLUSIONS: Our findings suggest that certain solutions may be more effective in removing bacteria from bone than mechanical irrigation with saline solution alone . Among the various solutions examined, the soap solution preserved the number and activity of osteoblasts the most . Low-pressure lavage with the soap solution resulted in the greatest removal of adherent bacteria from bone. Z Kardiol, 2001 Feb, 90(2), 133 - 7 {Aortic root abscess without involvement of the aortic valve: diagnosis and therapy in a 2.5-year-old child}; Hofbeck M et al.; Although formation of an aortic root abscess is a frequent complication of aortic valve endocarditis in adults, this complication has been rarely observed in children . In the majority of cases it has been described in children without underlying congenital heart disease . Due to the rarity of this complication, diagnosis and treatment is frequently delayed in childhood . We report a 2 1/2 year old girl who developed pericardial effusion in the course of pneumonia . Echocardiographic examinations, which were performed because of the pericardial effusion, revealed after 6 days the development of a cystic structure posterior to the aortic root . There was a perforation of this aortic root abscess to the left ventricular outflow tract; the aortic and mitral valves however were normal without endocarditic vegetations . Surgery was performed on the 10th day following a rapid increase in the size of the abscess . During surgery the abscess was drained and the perforation to the left ventricle was closed with direct sutures . Intraoperative transesophageal echocardiography confirmed a good surgical result . Blood cultures remained negative; in the material from the abscess however we found staphylococcus aureus . The postoperative course was uneventful . Our case demonstrates the necessity of detailed and repeated echocardiographic examinations in children with possible symptoms of bacterial endocarditis (in our case pericardial effusion) as well as the requirement of cultures of the abscess for identification of the infective organism . Intraoperative transesophageal echocardiography allows exact description of an aortic root abscess, its relation to other cardiac structures and immediate evaluation of the surgical result. Arch Orthop Trauma Surg, 2001, 121(3), 170 - 3 Effect of granulocyte-macrophage colony-stimulating factor on treatment of acute osteomyelitis . An experimental investigation in rats; Subasi M et al.; Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine that affects the various developmental steps of hematopoietic cells and enhances the phagocytic activity of these cells . The effect of GM-CSF on acute osteomyelitis, developed in rats, was investigated . For this purpose, osteomyelitis was firstly developed through the direct inoculation of Staphylococcus aureus into rat tibial metaphysis . Twenty-four rats in which diagnosis of osteomyelitis was histopathologically established were divided into two groups . Antibiotic only was given to the first group, and antibiotic as well as GM-CSF to the second group . Rats were followed up for 3 months with plain radiographs and scintigraphic methods using 67Ga-citrate . Material obtained from the rats that had been killed at the end of the 3rd month were histopathologically investigated . One rat in the first group died . In another rat, chronic osteomyelitis developed and fracture was observed . In 12 rats of the second group, physical examination, plain radiographs, and histopathologic findings were normal . In scintigraphic studies with 67Ga-citrate, when the pre- and posttreatment value of the same groups were evaluated by the Mann-Whitney U-test, the mean values at 48 h after treatment were found to be significant (P < 0.05), indicating a decrease in the 2nd group (experimental group) . In conclusion, the antibiotics were effective in the elimination of infection only together with neutrophils . In this manner, infections may be eliminated by strengthening the host's defense mechanism as well as by administering antibiotics . We believe that an adequate number of long-term studies will shed light on this issue . Besides we consider that this factor will be more important in the study of chronic osteomyelitis. Nat Immunol, 2000 Dec, 1(6), 533 - 40 Toll-like receptor 2-mediated NF-kappa B activation requires a Rac1-dependent pathway; Arbibe L et al.; Mammalian Toll-like receptors (TLRs) are expressed on innate immune cells and respond to the membrane components of Gram-positive or Gram-negative bacteria . When activated, they convey signals to transcription factors that orchestrate the inflammatory response . However, the intracellular signaling events following TLR activation are largely unknown . Here we show that TLR2 stimulation by Staphylococcus aureus induces a fast and transient activation of the Rho GTPases Rac1 and Cdc42 in the human monocytic cell line THP-1 and in 293 cells expressing TLR2 . Dominant-negative Rac1N17, but not dominant-negative Cdc42N17, block nuclear factor-kappa B (NF-kappa B) transactivation . S . aureus stimulation causes the recruitment of active Rac1 and phosphatidylinositol-3 kinase (PI3K) to the TLR2 cytosolic domain . Tyrosine phosphorylation of TLR2 is required for assembly of a multiprotein complex that is necessary for subsequent NF-kappa B transcriptional activity . A signaling cascade composed of Rac1, PI3K and Akt targets nuclear p65 transactivation independently of I kappa B alpha degradation . Thus Rac1 controls a second, I kappa B-independent, pathway to NF-kappa B activation and is essential in innate immune cell signaling via TLR2. Nat Immunol, 2000 Nov, 1(5), 441 - 6 Fc alpha/mu receptor mediates endocytosis of IgM-coated microbes; Shibuya A et al.; IgM is the first antibody to be produced in a humoral immune response and plays an important role in the primary stages of immunity . Here we describe a mouse Fc receptor, designated Fc alpha/microR, and its human homolog, that bind both IgM and IgA with intermediate or high affinity . Fc alpha/microR is constitutively expressed on the majority of B lymphocytes and macrophages . Cross-linking Fc alpha/microR expressed on a pro-B cell line Ba/F3 transfectant with soluble IgM or IgM-coated microparticles induced internalization of the receptor . Fc alpha/microR also mediated primary B lymphocyte endocytosis of IgM-coated Staphylococcus aureus . Thus, Fc alpha/microR is involved in the primary stages of the immune response to microbes. Biochemistry, 2001 Mar 13, 40(10), 3016 - 26 The antibacterial peptide pyrrhocoricin inhibits the ATPase actions of DnaK and prevents chaperone-assisted protein folding; Kragol G et al.; Recently, we documented that the short, proline-rich antibacterial peptides pyrrhocoricin, drosocin, and apidaecin interact with the bacterial heat shock protein DnaK, and peptide binding to DnaK can be correlated with antimicrobial activity . In the current report we studied the mechanism of action of these peptides and their binding sites to Escherichia coli DnaK . Biologically active pyrrhocoricin made of L-amino acids diminished the ATPase activity of recombinant DnaK . The inactive D-pyrrhocoricin analogue and the membrane-active antibacterial peptide cecropin A or magainin 2 failed to inhibit the DnaK-mediated phosphate release from adenosine 5'-triphosphate (ATP) . The effect of pyrrhocoricin on DnaK's other significant biological function, the refolding of misfolded proteins, was studied by assaying the alkaline phosphatase and beta-galactosidase activity of live bacteria . Remarkably, both enzyme activities were reduced upon incubation with L-pyrrhocoricin or drosocin . D-Pyrrhocoricin, magainin 2, or buforin II, an antimicrobial peptide involved in binding to bacterial nucleic acids, had only negligible effect . According to fluorescence polarization and dot blot analysis of synthetic DnaK fragments and labeled pyrrhocoricin analogues, pyrrhocoricin bound with a K(d) of 50.8 microM to the hinge region around the C-terminal helices D and E, at the vicinity of amino acids 583 and 615 . Pyrrhocoricin binding was not observed to the homologous DnaK fragment of Staphylococcus aureus, a pyrrhocoricin nonresponsive strain . In line with the lack of ATPase inhibition, drosocin binding appears to be slightly shifted toward the D helix . Our data suggest that drosocin and pyrrhocoricin binding prevents the frequent opening and closing of the multihelical lid over the peptide-binding pocket of DnaK, permanently closes the cavity, and inhibits chaperone-assisted protein folding . The biochemical results were strongly supported by molecular modeling of DnaK-pyrrhocoricin interactions . Due to the prominent sequence variations of procaryotic and eucaryotic DnaK molecules in the multihelical lid region, our findings pave the road for the design of strain-specific antibacterial peptides and peptidomimetics . Far-fetched applications of the species-specific inhibition of chaperone-assisted protein folding include the control of not only bacteria but also fungi, parasites, insects, and perhaps rodents. Biochemistry, 2001 Mar 6, 40(9), 2964 - 71 Regiospecificity assignment for the reaction of kanamycin nucleotidyltransferase from Staphylococcus aureus; Gerratana B et al.; Aminoglycoside nucleotidyltransferases catalyze the transfer of a nucleoside monophosphoryl group from a nucleotide to a hydroxyl group of an aminoglycoside antibiotic . Kanamycin nucleotidyltransferase {ANT (4',4' ')-I} from Staphylococcus aureus confers resistance to numerous aminoglycosides with a 4' or 4' ' hydroxyl group in the equatorial position . The synthesis of m-nitrobenzyl triphosphate, a new substrate of kanamycin nucleotidyltransferase, is reported . The kanamycin nucleotidyltransferase catalyzed reaction of kanamycin A with m-nitrobenzyl triphosphate is 2 orders of magnitude slower than that with ATP . The MALDI-TOF spectra of the purified products of both reactions revealed that kanamycin A was modified only at one position . The regiospecificity of the reaction catalyzed by kanamycin nucleotidyltransferase of kanamycin A with either ATP or m-nitrobenzyl triphosphate was determined directly by one- and two-dimensional hetero- and homonuclear NMR techniques . The site of the modification was unambiguously assigned to the 4' hydroxyl of kanamycin A; thus, the products formed are 4'-(adenosine-5'-phosphoryl)-kanamycin A and 4'-(m-nitrobenzyl phosphoryl)-kanamycin A . This eliminates the uncertainty concerning the point of modification since this could not be determined from the crystal structure of the enzyme with bound MgAMPCPP and kanamycin A {Pedersen, L . C., Benninig, M . M., and Holden, H . M . (1995) Biochemistry 34, 13305-13311}. Biotechnol Bioeng, 2001 May 5, 73(3), 246 - 52 Improving biomaterial properties of collagen films by chemical modification; Tiller JC et al.; Films of bovine collagen were chemically modified with the goal of improving their biomaterial properties . The modified films were investigated with respect to their affinity to fibroblast and endothelial cells, as well as their antibacterial properties tested by adhesion of Staphylococcus aureus . Modifications that only change the net charge of collagen, such as acetylation, succinylation, and treatment with glutaraldehyde (all increase the negative charge), and amination with ethylenediamine (EDA), N,N-dimethyl-EDA (DMEDA), or butylamine (all increase the positive charge), did not dramatically alter the mammalian cell attachment to the film . In contrast, derivatization of collagen using methoxypoly(ethylene glycol) (PEG) diminished the attachment of fibroblasts by 98 +/- 1% and of endothelial cells by more than 99% compared to unmodified collagen . Moreover, the rate of growth of fibroblasts dropped by 97 +/- 1% and that of endothelial cells by 88 +/- 3% as a result of PEGylation of collagen . Adhesion of S . aureus cells also plummeted by 93 +/- 2% as a result of this PEGylation . With these antifouling properties, PEG-collagen may be a promising coating material for coronary stents . Subsequent derivatization of PEG-collagen with EDA or DMEDA abolished its mammalian cell-repelling ability, whereas bacterial cell repulsion was partially retained: for example, DMEDA-modified PEG-collagen exhibits up to a 5-fold lower bacterial adhesion than collagen . It is worth noting that a material that allows mammalian cell attachment but reduces bacterial adhesion could be useful as an implant or coating . Vaccine, 2001 Mar 21, 19(17-19), 2542 - 8 The B cell targeted adjuvant, CTA1-DD, exhibits potent mucosal immunoenhancing activity despite pre-existing anti-toxin immunity; Lycke N et al.; We recently developed a novel immunomodulating gene fusion protein, CTA1-DD, that combines the ADP-ribosylating ability of cholera toxin (CT) with a dimer of an Ig-binding fragment, D, of Staphylococcus aureus protein A . The CTA1-DD adjuvant was found to be non-toxic and greatly augmented T cell dependent and independent responses . Following injection it binds to both naive and memory B cells and up-regulates co-stimulatory molecules as well as prevents apoptosis of activated B cells . Here we show that CTA1-DD is a potent mucosal adjuvant administered intranasally . A dose-response analysis revealed that the adjuvant effect of CTA1-DD given intranasally was equally strong to that observed after systemic immunizations . The adjuvant effect was independent of any possible contamination with endotoxin as indicated by the similar enhancing effects of CTA1-DD in C3H/HeN and the LPS-insensitive C3H/HeJ mice . Contrary to many other adjuvants CTA1-DD induces an immune response to itself . However, despite the presence of high serum titers of pre-existing anti-CTA1 antibodies we observed no reduction of the adjuvant function of CTA1-DD when given either intranasally or systemically . These results support the notion that the CTA1-DD adjuvant can repeatedly be used in the clinic without loss of efficacy even when pre-existing anti-CTA1 antibody levels are high. Antimicrob Agents Chemother, 2001 Apr, 45(4), 1292 - 4 Combination effect of vancomycin and beta-lactams against a Staphylococcus aureus strain, Mu3, with heterogeneous resistance to vancomycin; Aritaka N et al.; We tested the combined activity of vancomycin and seven beta-lactam antibiotics against Staphylococcus aureus clinical strain Mu3, which displays heterogeneous resistance to vancomycin . When combined with vancomycin, four of the seven tested beta-lactams exhibited an additive effect at or near their MICs, while all showed an antagonistic effect at lower, sub-MIC levels . This study implicated the unpredictable nature of combination therapy of beta-lactams and vancomycin against S . aureus with reduced susceptibility to vancomycin. Antimicrob Agents Chemother, 2001 Apr, 45(4), 1244 - 8 Activities of the combination of quinupristin-dalfopristin with rifampin in vitro and in experimental endocarditis due to Staphylococcus aureus strains with various phenotypes of resistance to macrolide-lincosamide-streptogramin antibiotics; Zarrouk V et al.; We evaluated the activities of quinupristin-dalfopristin (Q-D), alone or in combination with rifampin, against three strains of Staphylococcus aureus susceptible to rifampin (MIC, 0.06 microg/ml) and to Q-D (MICs, 0.5 to 1 microg/ml) but displaying various phenotypes of resistance to macrolide-lincosamide-streptogramin antibiotics: S . aureus HM1054 was susceptible to quinupristin and dalfopristin (MICs of 8 and 4 microg/ml, respectively); for S . aureus RP13, the MIC of dalfopristin was high (MICs of quinupristin and dalfopristin for strain RP13, 8 and 32 microg/ml, respectively); and S . aureus HM1054R was obtained after conjugative transfer of macrolide-lincosamide-streptogramin B constitutive resistance to HM1054, and the MIC of quinupristin for this strain was high (MICs of quinupristin and dalfopristin, 64 and 4 microg/ml, respectively) . In vitro time-kill curve studies showed an additive effect {corrected} between Q-D and rifampin, at a concentration of four times the MIC, against the three strains . Rabbits with aortic endocarditis were treated 4 days with Q-D, rifampin, or their combination . In vivo, the combination was highly bactericidal and synergistic against strains susceptible to quinupristin (HM1054 and RP13) and sterilized 94% of the animals . In contrast, the combination was neither synergistic nor bactericidal against the quinupristin-resistant strain (HM1054R) and did not prevent the emergence of mutants resistant to rifampin . We conclude that the in vivo synergistic and bactericidal activity of the combination of Q-D and rifampin against S . aureus is predicted by the absence of resistance to quinupristin but not by in vitro combination studies. Clin Sci (Lond), 2001 Apr, 100(4), 395 - 400 A novel method for the delivery of nitric oxide therapy to the skin of human subjects using a semi-permeable membrane; Hardwick JB et al.; Nitric oxide (NO) is a mediator of essential biological processes, including vasodilatation, anti-microbial activity and wound healing . A chemical system using sodium nitrite and ascorbic acid has been developed which generates significant amounts of NO . The originally described system was messy and impractical, and the high acidity may cause pain and further tissue damage in ulcerated skin . To overcome this, a selectively permeable, hydrophilic polyester co-polymer membrane system (Sympatex ) has been identified that can be placed between the NO-generating chemicals and the skin . The aim of the present study was to determine whether NO derived from this chemical system was able to diffuse through the membrane and have a measurable vasodilatory effect on forearm skin in healthy volunteers . The Sympatex 10 microm membrane was found to be highly permeable to NO, while preventing passage of the constituents of the NO-generation gel to the skin . The transmembrane NO-generation system had a vasodilatory effect comparable with that resulting from direct topical application . Additionally, the NO generated was effective in killing Staphylococcus aureus and Escherichia coli at doses lower than those required to increase skin blood flow . The vasodilatory and anti-microbial effects of this system may be useful as a patch-based topical therapy for skin ulceration, particularly when there is concomitant ischaemia and infection. Arch Dermatol, 2001 Mar, 137(3), 313 - 5 Antibiotic prophylaxis for full-face laser resurfacing: is it necessary? Gaspar Z, Vinciullo C, Elliott T. OBJECTIVE: To evaluate the need for antibiotic prophylaxis when performing full-face laser resurfacing . METHOD: Prospective study of 31 patients undergoing full-face laser resurfacing, 17 with and 14 without antibiotic prophylaxis . OBSERVATION: Four of 14 patients without antibiotic prophylaxis had microbiologic and clinical evidence of infection . None of the 17 patients with antibiotic prophylaxis had clinical infection . Early treatment prevented adverse sequelae in the 4 patients who developed infection . CONCLUSION: Antibiotic prophylaxis against Staphylococcus aureus is useful but not essential, because meticulous wound care and close clinical monitoring of patients daily with routine bacterial swabs can detect infection early. J Biomed Mater Res, 2001 May, 55(2), 217 - 28 In vivo response to biodegradable controlled antibiotic release systems; Korkusuz F et al.; In this study, the major goal was to evaluate in vitro and in vivo findings by macroscopy, radiology, and histology to determine the effectiveness of therapy of experimental implant-related osteomyelitis with antibiotic carrier rods constructed of microbial polyesters . The polymers used were poly(3-hydroxybutyrate-co-4-hydroxyvalerate) {P(3-HB-co-4-HB)} and poly(3-hydroxybutyrate-co-3-hydroxy- valerate) {P(3-HB-co-3-HV)} . Both the Sulperazone and the Duocid-P(3-HB-co-4-HB) rods with a drug to polymer ratio of 1:1 (w/w) were effective in treating the bone infection that was experimentally initiated by inoculation of a hemolytic strain of Staphylococcus aureus (coagulase positive; phage type 52/52b) together with metal implants into the medullary area of rabbit tibia . Macroscopical data revealed that the effectiveness of therapy was apparent at week 6 for all categories tested . Radiological findings with Duocid- and Sulperazone-loaded P(3-HB-co-4-HB) rods improved significantly when judged by changes in periosteal elevation, widening of bone shaft, new bone formation, and soft-tissue deformation after 6 weeks of implantation . Histologically the signs of infection were found to subside by weeks 3 and 6 . Inflammatory cells were replaced with bone-forming cells upon treatment with Sulperazone-P(3-HB-co-4-HB) and Duocid-P(3-HB-co-4-HB) . Osteoblastic activity was prominent . Intramedullary inflammation, although still present, started to be replaced by fibrous or bony tissue . Histological findings presented the subsidence of infection . In summary, the antibiotic-loaded biopolymeric rods appeared to have potential as a new controlled-release system for the treatment of implant related osteomyelitis and chronic osteomyelitis . J Cataract Refract Surg, 2001 Mar, 27(3), 471 - 3 Methicillin-resistant Staphylococcus aureus keratitis after laser in situ keratomileusis; Rudd JC et al.; A 50-year-old man had uneventful bilateral laser in situ keratomileusis (LASIK) for moderate myopia (-4.50 diopters sphere, both eyes) . Twelve days postoperatively, he developed unilateral bacterial keratitis . Cultures revealed methicillin-resistant Staphylococcus aureus . The antibiotic regimen was adjusted, and he regained an uncorrected visual acuity of 20/40 and a best spectacle-corrected visual acuity (BSCVA) of 20/15 . Bacterial keratitis after LASIK is a rare occurrence . Aggressive use of cultures and fortified antibiotics can prevent significant loss of BSCVA, even when a resistant organism is the cause. J Immunol, 2001 Apr 1, 166(7), 4634 - 43 CXC chemokine receptor-2 ligands are required for neutrophil-mediated host defense in experimental brain abscesses; Kielian T et al.; We have developed a mouse brain abscess model by using Staphylococcus aureus, one of the main etiologic agents of brain abscesses in humans . Direct damage to the blood-brain barrier was observed from 24 h to 7 days after S . aureus exposure as demonstrated by the accumulation of serum IgG in the brain parenchyma . Evaluation of brain abscesses by immunohistochemistry and flow cytometry revealed a prominent neutrophil infiltrate . To address the importance of neutrophils in the early containment of S . aureus infection in the brain, mice were transiently depleted of neutrophils before implantation of bacteria-laden beads . Neutrophil-depleted animals consistently demonstrated more severe brain abscesses and higher CNS bacterial burdens compared with control animals . S . aureus led to the induction of numerous chemokines in the brain, including macrophage-inflammatory protein (MIP)-1alpha/CCL3, MIP-1beta/CCL4, MIP-2/CXCL1, monocyte chemoattractant protein-1/CCL2, and TCA-3/CCL1, within 6 h after bacterial exposure . These chemokines also were expressed by both primary cultures of neonatal mouse microglia and astrocytes exposed to heat-inactivated S . aureus in vitro . Because neutrophils constitute the majority of the cellular infiltrate in early brain abscess development, subsequent analysis focused on MIP-2 and KC/CXCL1, two neutrophil-attracting CXC chemokines . Both MIP-2 and KC protein levels were significantly elevated in the brain after S . aureus exposure . Neutrophil extravasation into the brain parenchyma was impaired in CXCR2 knockout mice and was associated with increased bacterial burdens . These studies demonstrate the importance of the CXCR2 ligands MIP-2 and KC and neutrophils in the acute host response to S . aureus in the brain. Infect Immun, 2001 Apr, 69(4), 2448 - 55 SarS, a SarA homolog repressible by agr, is an activator of protein A synthesis in Staphylococcus aureus; Cheung AL et al.; The expression of protein A (spa) is repressed by global regulatory loci sarA and agr . Although SarA may directly bind to the spa promoter to downregulate spa expression, the mechanism by which agr represses spa expression is not clearly understood . In searching for SarA homologs in the partially released genome, we found a SarA homolog, encoding a 250-amino-acid protein designated SarS, upstream of the spa gene . The expression of sarS was almost undetectable in parental strain RN6390 but was highly expressed in agr and sarA mutants, strains normally expressing high level of protein A . Interestingly, protein A expression was decreased in a sarS mutant as detected in an immunoblot but returned to near-parental levels in a complemented sarS mutant . Transcriptional fusion studies with a 158- and a 491-bp spa promoter fragment linked to the xylE reporter gene disclosed that the transcription of the spa promoter was also downregulated in the sarS mutant compared with the parental strain . Interestingly, the enhancement in spa expression in an agr mutant returned to a near-parental level in the agr sarS double mutant but not in the sarA sarS double mutant . Correlating with this divergent finding is the observation that enhanced sarS expression in an agr mutant was repressed by the sarA locus supplied in trans but not in a sarA mutant expressing RNAIII from a plasmid . Gel shift studies also revealed the specific binding of SarS to the 158-bp spa promoter . Taken together, these data indicated that the agr locus probably mediates spa repression by suppressing the transcription of sarS, an activator of spa expression . However, the pathway by which the sarA locus downregulates spa expression is sarS independent. Clin Rheumatol, 2001, 20(1), 10 - 4 A comparison between septic bursitis caused by Staphylococcus aureus and those caused by other organisms; Cea-Pereiro JC et al.; Septic bursitis is an infection that usually involves olecranon and prepatellar bursae . Staphylococcus aureus is responsible for around 80% of cases . However, information regarding bursitis caused by non-Staphylococcus aureus microorganisms (NSAB) is scant . In this paper we describe the characteristics of NSAB and emphasise differences between these and Staphylococcus aureus bursitis (SAB) . A retrospective study of all cases with septic bursitis seen between January 1991 and June 1998 at one university hospital was conducted . Only cases in which bursal fluid culture yielded growth of a microorganism were analysed . A literature review was conducted for completeness . Fifty-seven episodes of septic bursitis in 56 patients were studied: 47 of these were caused by Staphylococcus aureus and 11 by non-Staphylococcus aureus microorganisms . Forty-three SAB patients were male (91%) . Mean age at diagnosis was 50 years (range 20-85 years) . The presentation of bursitis had a seasonal trend, with a peak in the summer . Twenty-three patients (51%) had occupations involving frequent or sustained pressure on the bursae . Other risk factors were recent trauma in 11 (23%), alcoholism in six (13%), pre-existing bursal disease in five (11%), and chronic obstructive pulmonary disease in four (9%) . There were 20 cases of olecranon bursitis (43%), 25 of prepatellar bursitis (53%) and two of first metatarsophalangeal bursitis . Characteristics of patients from the literature review were similar . Eight NSAB patients (73%) were male . Mean age at diagnosis was 46.9 (range 29-83 years) . Two patients were plumbers and one a stonemason . Five (45%) had neither putative systemic nor local risk factors . There were five olecranon (45%), five prepatellar (45%), and one external malleolus bursitis . Infection by a mixed flora was common . Unlike SAB, the presentation of cases did not have a seasonal trend . The clinical spectrum of non-Staphylococcus aureus bursitis (NSAB) differs from that of Staphylococcus Aureus bursitis (SAB), and this should be considered in the initial diagnosis of septic bursitis. J Vet Med B Infect Dis Vet Public Health, 2001 Feb, 48(1), 21 - 9 Comparison of methods for the determination of antimicrobial resistance in Staphylococcus aureus from bovine mastitis; Schlegelova J et al.; The results of three standard methods (broth dilution, agar dilution, disk diffusion) and an experimental modification of the microdilution method for determination of resistance to ampicillin, cephalotin, cloxacillin, neomycin, novobiocin, penicillin and streptomycin were compared using 151 Staphylococcus aureus isolates obtained from cases of mastitis . The accuracy of the dilution methods was compared by determination of minimum inhibition concentrations (MIC, MIC50, MIC90 and modal MIC) and by assessment of the agreement within the tolerance of +/-1 dilution step in 2-fold dilution series . The results of the dilution methods were further compared with those of the reference disk diffusion method and the strains were classified as sensitive or resistant using the interpretation criteria for human strains . The comparisons indicated that MIC characteristics and the final classification as sensitive or resistant were method-dependent . Resistance to aminoglycoside antibiotics was observed more often when using broth dilution methods, especially when the broth was supplemented with lactose. J Antimicrob Chemother, 1999 Feb, 43(2), 291 - 4 In-vitro bactericidal activity of cefpirome and cefamandole in combination with glycopeptides against methicillin-resistant Staphylococcus aureus; Bergeret M et al.; The bactericidal activity in vitro of cefpirome plus either vancomycin or teicoplanin was compared with that of a cefamandole-vancomycin combination against ten clinical isolates of homogeneous methicillin-resistant Staphylococcus aureus . Cefpirome (0.125 x MIC) combined with vancomycin (0.5-2 x MIC) or teicoplanin (0.5-4 x MIC) acted synergically against the ten isolates . Similar effects were observed with the cefamandole-vancomycin combination, except that for one isolate, higher cefamandole concentrations (0.25-1 x MIC) were required. Clin Infect Dis, 2001 Mar 15, 32 Suppl 1, S16 - 22 Comparative in vitro and in vivo activity of the C-8 methoxy quinolone moxifloxacin and the C-8 chlorine quinolone BAY y 3118; Dalhoff A; The C-8 methoxy quinolone moxifloxacin is highly bactericidal against wild-type and first-step gyrase- and topoisomerase IV-resistant mutants . This finding led to the hypothesis that the C-8 methoxy group may lower the propensity for resistance development compared with quinolones possessing different substituents at the C-8 position . Therefore, resistance development of the C-8 methoxy quinolone moxifloxacin was compared with that of its structural analogue BAY y 3118 (chlorine moiety at the C-8 position), with Staphylococcus aureus used as the test organism . The spontaneous emergence of resistance was quantified by counting the number of colonies growing on drug-free medium compared with moxifloxacin- or BAY y 3118-containing media . The multistep emergence of quinolone resistance was encountered by growing S . aureus over 8 passages in drug-containing medium . Human serum concentrations were simulated in an in vitro model over 84 h (dosing every 24 h), and total and resistant S . aureus were quantified . Spontaneous mutation frequencies of 6x10-11 for moxifloxacin and 4x10-7 for BAY y 3118 were observed . Multistep resistance to moxifloxacin developed slowly (2-fold rise) but rapidly against BAY y 3118 (>16-fold rise) . No resistance against moxifloxacin developed in this model, whereas resistance to BAY y 3118 began to develop after 4 h . Thus, as the C-8 moiety was the only difference, the 8-methoxy group on moxifloxacin appeared to significantly lower the propensity for quinolone resistance development. Curr Opin Mol Ther, 2001 Feb, 3(1), 37 - 44 The B-cell targeted CTA1-DD vaccine adjuvant is highly effective at enhancing antibody as well as CTL responses; Lycke N; A novel immunomodulating gene-fusion protein, CTA1-DD, has been developed which combines the ADP-ribosylating ability of cholera toxin (CT) with a dimer of an Ig-binding fragment, D, of Staphylococcus aureus protein A . The CTA1-DD adjuvant is non-toxic and greatly augmented T-cell-dependent and T-cell-independent responses to soluble admixed antigens after systemic as well as mucosal immunizations . CTL and antibody responses of all classes were increased by 10- to 100-fold above those observed in control mice immunized without adjuvant . CTA1-DD does not appear to form immune complexes or bind to soluble Ig following injection, but rather it binds directly to B-cells of all isotypes, including naive IgD+ cells . No binding was observed to macrophages or dendritic cells, and immunizations in Fc gamma R-deficient mice demonstrated unaltered enhancing effects . As shown by inactive mutants, the CTA1-DD adjuvant is dependent on ADP-ribosyltransferase activity and requires the binding to Ig- via the DD moiety . The enhancing effect is associated with enlarged germinal centers, and binding of CTA1-DD to the B-cells strongly upregulates co-stimulatory molecules and counteracts apoptosis by inducing intracellular Bcl-2. Clin Infect Dis, 2001 Mar 15, 32(6), 877 - 83 Epub 2001 Mar 09. Poststernotomy mediastinitis due to Staphylococcus aureus: comparison of methicillin-resistant and methicillin-susceptible cases; Mekontso-Dessap A et al.; The objective of the study was to compare the outcome of poststernotomy mediastinitis (PSM) caused by methicillin-resistant and methicillin-susceptible Staphylococcus aureus (MRSA and MSSA, respectively) . Hospital records of 41 patients with S . aureus PSM who were all treated by closed drainage from 1 April 1996 through 1 February 2000 were reviewed . PSM was caused by MRSA in 15 patients and by MSSA in 26 . Follow-up (+/-SD) averaged 12.5+/-14.0 months per patient . Both groups had similar perioperative characteristics . Patients with MRSA PSM had a significantly lower actuarial survival rate than did patients with MSSA PSM (60.0%+/-12.6%, 52.5%+/-3.4%, and 26.3%+/-19.7% versus 84.6%+/-7.1%, 79.0%+/-8.6%, and 79.0%+/-8.65 at 1 month, and at 1 and 3 years, respectively; values are +/- SD; P=.04) . PSM-related death and treatment failure were significantly higher in the MRSA group than in the MSSA group (P=.03 and.02, respectively) . Logistic regression analysis revealed that MRSA was the only independent risk factor for overall mortality . In conclusion, the clinical outcome of PSM caused by MRSA is poorer than that caused by MSSA. Drug Dev Ind Pharm, 2001 Jan, 27(1), 97 - 101 Synthesis and properties of dextran-linked ampicillin; Kim DS et al.; Ampicillin was coupled to dextran of average molecular weight 9,000 or 81,200 via the cyanogen bromide method . The degree of drug substituted per glucose unit (DSG) was varied from 0.104 to 0.028 (by weight bases: 22.1-5.9%) depending on the ratio of the reactants . Water solubility of dextran-linked ampicillin increased compared with free ampicillin, and the solubility decreased as the amount of ampicillin substituted increased . Plasma concentration of ampicillin, which appeared after intravenous administration of dextran-linked ampicillin in rats, was higher than when free ampicillin was administered, and the more so, the higher the molecular weight of dextran . Plasma half-life of dextran-linked ampicillin was two times longer than that of free ampicillin in rats . Antibacterial activities of dextran-linked ampicillin were evaluated against Staphylococcus aureus, Bacillus substillis, and Escherichia coli at two concentration levels according to the cup-plate method by measuring the diameter of inhibition zone, which was comparable to that of free ampicillin. Indian J Med Res, 2000 Dec, 112, 198 - 202 Comparison of various conventional methods with a polymerase chain reaction assay for detecting methicillin-resistant & susceptible Staphylococcus aureus strains; Prasad KN et al.; BACKGROUND & OBJECTIVES: Methicillin-resistant Staphylococcus aureus (MRSA), a major nosocomial pathogen world-wide, is often difficult to detect due to the heterogeneous nature of expression of oxacillin resistance . In the present study, various conventional methods were compared with polymerase chain reaction on 106 clinical isolates of Staph . aureus for detection of oxacillin resistance . METHODS: A total of 106 clinical isolates of Staph . aureus were tested for oxacillin resistance by disc diffusion, screen agar plates (3 micrograms and 6 micrograms/ml of oxacillin), oxacillin broth (3 micrograms/ml) and mecA based PCR . RESULTS: PCR detected mecA gene amplified product of 604 bp in 57 strains . Disc diffusion failed to detect 7 mecA positive strains but identified 5 mecA negative strains as oxacillin resistant . Screen agar 3 micrograms, screen agar 6 micrograms and oxacillin broth 3 micrograms detected 55, 53 and 55 respectively of the 57 mecA positive strains; however, they also falsely identified 5, 3 and 3 strains of mecA negative strains respectively as oxacillin resistant . The sensitivity, specificity and accuracy of disc diffusion, 3 micrograms screen agar, 6 micrograms screen agar and 3 micrograms oxacillin broth against PCR as gold standard were as follows: 87.7, 89.9 and 88.7 per cent; 96.5, 89.8 and 93.4 per cent; 93.0, 93.9 and 93.4 per cent; 96.5, 93.9 and 95.3 per cent respectively . INTERPRETATION & CONCLUSIONS: The present study demonstrated that disc diffusion test was least reliable and 3 micrograms broth had the highest sensitivity and specificity when compared with PCR for detection of oxacillin resistance . Because of variations among the methods, a combination of tests should be used for the accurate detection of MRSA till new guidelines by an appropriate body are formulated. J Bone Joint Surg Br, 2001 Jan, 83(1), 93 - 8 Primary subacute haematogenous osteomyelitis in children; Rasool MN; Between 1990 and 1998 we saw 21 children with primary subacute haematogenous osteomyelitis . Pain, swelling and a limp had been present for two to 12 weeks with little functional impairment . Laboratory tests were non-contributory . The lesions were classified radiologically into metaphyseal, diaphyseal, epiphyseal and vertebral . There were 24 sites involved, with most (20) being in the tibia; 17 lesions were in the diaphysis, five in the metaphysis and two in the epiphysis . The diagnosis was confirmed histologically in all cases . Staphylococcus aureus was cultured in six patients . Healing occurred in all patients after treatment with antibiotics for six weeks and radiological improvement was seen after three to six months . Subacute osteomyelitis develops as a result of increased host resistance and decreased bacterial virulence . The radiological features can mimic various benign or malignant bone tumours and non-pyogenic infections . Histological confirmation is necessary to avoid a delay in diagnosis. Eur J Clin Microbiol Infect Dis, 2001 Jan, 20(1), 27 - 32 Highly epidemic strains of methicillin-resistant Staphylococcus aureus not distinguished by capsule formation, protein A content or adherence to HEp-2 cells; Aathithan S et al.; In order to investigate whether highly epidemic methicillin-resistant Staphylococcus aureus (EMRSA) strains possess special properties that favour their dissemination and survival, a study was undertaken that examined methicillin-sensitive and methicillin-resistant strains of Staphylococcus aureus isolated in the UK . Included in the study were EMRSA types 1, 2, 3, 15 and 16 . Phage types EMRSA-15 and -16, in particular, have emerged as significant hospital pathogens in the UK, resisting standard methods of control and spreading widely, while the incidence of other epidemic types has either declined or not changed . All of the strains included in the study were examined for capsule formation, the amount of bound protein A produced, and quantitative adherence to the human continuous epithelial cell line HEp-2 . Although all of these properties varied amongst the strains examined, there was no relationship between any of them and methicillin resistance or epidemic type and, incidentally, no relationship between cell wall-bound protein A content and adherence. ORL J Otorhinolaryngol Relat Spec, 2001 Mar-Apr, 63(2), 87 - 91 Possibility of reciprocal infection of methicillin-resistant Staphylococcus aureus between medical personnel and patients undergoing middle ear surgery; Lee ES et al.; The incidence of methicillin-resistant Staphylococcus aureus (MRSA) infection after middle ear surgery has recently increased in our hospital . In this study we tried to determine whether the strains of MRSA isolated from infected patients are identical to those obtained from medical personnel, to prove a reciprocal transmission between medical personnel and patients . Surveillance bacterial cultures of medical personnel were performed from the anterior nares and from the fingertip . Molecular epidemiological studies, ribotyping and pulsed-field gel electrophoresis (PFGE) were used to compare the 12 MRSA strains obtained from carriers among medical personnel with the 60 strains identified from patient's otorrhea . Six different MRSA strains were identified from ribotyping, and three subtypes from PFGE . There was a particular subtype which was the most frequently identified strain found in both medical carriers and patients . Postoperative MRSA infection rates after the treatment of medical carriers and application of preventive procedures decreased from 11.9 to 5.7% . These findings suggest that the MRSA transmissions have occurred between medical personnel and patients . Nephron, 2001 Feb, 87(2), 177 - 81 Reversible nephrotic syndrome in a patient with amyloid A amyloidosis of the kidney following methicillin-resistant Staphylococcus aureus infection; Yokota N et al.; A common form of methicillin-resistant Staphylococcus aureus (MRSA) associated glomerulonephritis is either an endocapillary proliferative glomerulonephritis or a crescentic glomerulonephritis . This report describes the development of reversible nephrotic syndrome following MRSA infection in a patient with amyloid A amyloidosis . The patient had been diagnosed as having rheumatoid arthritis for 50 years . Suppurative arthritis due to MRSA became complicated 2 years prior to admission to our hospital . In the meantime, a nonnephrotic-range proteinuria developed . Two weeks before admission, nephrotic syndrome developed . The serum creatinine level remained unchanged throughout the course, but common features characteristic of MRSA-associated glomerulonephritis were observed in this patient, such as elevated serum IgG and IgA levels . A renal biopsy specimen showed glomerular amyloid A amyloidosis of a nodular type, infiltrated mononuclear cells in the mesangium, deposition of IgG, IgA, and C3, and swelling of glomerular endothelial cells . There were no crescentic glomeruli . Following surgical eradication of the MRSA focus in the right knee joint, nephrotic syndrome disappeared . Hence, it was highly possible that MRSA infection induced a reversible nephrotic syndrome by causing reversible injuries to glomerular endothelial cells . The description of this case serves to illustrate the range of MRSA infections that may cause various forms of glomerulonephritides . J Bacteriol, 2001 Apr, 183(7), 2367 - 71 A mutation in the 5' untranslated region increases stability of norA mRNA, encoding a multidrug resistance transporter of Staphylococcus aureus; Fournier B et al.; NorA, a multidrug efflux pump in Staphylococcus aureus, protects the cell from multiple drugs, including quinolones . The flqB mutation (T-->G) in the 5' untranslated region upstream of norA causes norA overexpression of 4.9-fold in cis, as measured in norA::blaZ fusions . The transcriptional initiation site of norA was unchanged in mutant and wild-type strains, but the half-life of norA mRNA was increased 4.8-fold in the flqB mutant compared to the wild-type strain . Computer-generated folding of the first 68 nucleotides of the norA transcript predicts an additional stem-loop and changes in a putative RNase III cleavage site in the flqB mutant. Vet Res Commun, 2001 Feb, 25(2), 117 - 26 A comparison of the concentrations of C-reactive protein and alpha1-acid glycoprotein in the serum of young and adult dogs with acute inflammation; Hayashi S et al.; The concentrations of C-reactive protein (CRP) and alpha1-acid glycoprotein (AAG) were evaluated in 1-, 3- and 18-month-old dogs (four of each age) that had been inoculated with turpentine oil . The CRP and AAG in 3-month-old and younger dogs subjected to surgery or inoculated with either Staphylococcus aureus or a viral vaccine were also evaluated . The average CRP concentration in the sera peaked 2 days after inoculation of turpentine oil . The peak CRP concentrations in 3- and 18-month-old dogs were significantly (p < 0.05) greater than those in 1-month-old dogs . The average AAG concentration in the sera peaked 4 days after inoculation of turpentine oil . No significant difference was found in AAG concentrations between any of the age groups . When experimentally inoculated with S . aureus or subjected to oophorohysterectomy, the CRP and AAG concentrations increased in 3-month-old dogs, but they increased little in 1-month-old dogs . The CRP and AAG in dogs inoculated with the viral vaccine did not increase . In dogs with fractures or subjected to percutaneous gastrostomy, the CRP and AAG concentrations correlated with the condition of dogs. J Vet Diagn Invest, 2001 Jan, 13(1), 74 - 6 Pulmonary botryomycosis in a Scottish highland steer; Miller MA et al.; Pertinent necropsy findings in a 5 1/2-year-old Scottish Highland steer with chronic intractable pneumonia and cough were limited to the pulmonary system . The accessory lobe of the lung was collapsed, scarred, and multifocally adhered to parietal pleura . A polypoid mass almost completely obstructed the lobar bronchus and protruded into the trachea; mucopurulent exudate distended more distal bronchi . Botryomycosis was diagnosed when histologic examination revealed pyogranulomatous pneumonia with eosinophilic granules and "club" formation surrounding colonies of gram-positive cocci . Staphylococcus aureus was cultured from the lung . Botryomycosis is an unusual response to infection with common bacteria and is characterized by pyogranulomatous inflammation with formation of eosinophilic granules surrounding colonies of gram-positive cocci or gram-negative bacilli . Among domestic species, staphylococcal botryomycosis is most common as a wound infection in horses or as mastitis in cows and sows . Pulmonary botryomycosis is rare in horses, humans, and guinea pigs and apparently has not been reported in cattle. Spine, 2001 Mar 1, 26(5), 588 - 9 Pyogenic spondylitis in an S1-S2 immobile segment; Koh YD et al.; STUDY DESIGN: A case of pyogenic spondylitis in S1-S2 is presented . OBJECTIVE: To describe the diagnosis and management of this rare spondylitis . SUMMARY OF BACKGROUND DATA: The segment including the first and second sacral vertebrae is not mobile . Therefore, discitis of S1-S2 and adjacent spondylitis is very rare . To the authors' knowledge, this is the first reported case of infectious spondylitis in an immobile segment: S1-S2 . METHODS: In addition to radiography and bone scintigraphy, magnetic resonance imaging was used to confirm the diagnosis . Changes consistent with infectious spondylitis were shown, including an epidural abscess . RESULTS: The patient was treated with laparoscopic drainage and biopsy . Staphylococcus aureus was cultured, and adequate antibiotics were administered . Repeat magnetic resonance imaging at approximately 4 months demonstrated normal signal intensity and disappearance of the abscess . CONCLUSION: Findings from this study show that pyogenic spondylitis can occur in immobile S1-S2. Cytometry, 2001 Feb 15, 46(1), 33 - 40 Flow cytometric analysis of cytokine production by normal human peripheral blood dendritic cells and monocytes: comparative analysis of different stimuli, secretion-blocking agents and incubation periods; Bueno C et al.; In this paper, we comparatively analyze the effects of the following different stimuli on the production and intracellular accumulation of the interleukin (IL)-1 beta, IL-6, IL-12, tumor necrosis factor-alpha (TNF-alpha), and IL-8 inflammatory cytokines in both normal human peripheral blood (PB) dendritic cell (DC) subsets and monocytes: lipopolysaccharide (LPS) versus Staphylococcus aureus cowan I (SAC) in the presence or absence of interferon-(IFN)-gamma-, cytokine secretion-blocking agents (brefeldin A alone versus brefeldin A plus monensin), and incubation periods (6, 12, and 24 h) . For this purpose, a four-color multiple-staining direct immunofluorescence technique analyzed by flow cytometry was systematically used in all experiments (n = 19) . Our results show that after stimulation, an important proportion of each of the two CD33(+) myeloid DC subsets as well as the monocytes produce significant amounts of all cytokines analyzed under each of the experimental conditions assayed . In contrast, CD33(-/+lo) lymphoplasmocytoid DC failed to produce detectable levels of any of the above-mentioned cytokines under the same stimulatory conditions . Upon comparing the different stimuli used, LPS was associated with higher percentages of cytokine-producing cells compared with SAC, especially within the CD33(hi) DC subset; interestingly, the addition of IFN-gamma enhanced the response of monocytes to both LPS and SAC . As regards the secretion-blocking agents, brefeldin A alone was superior to the combination of brefeldin A and monensin . This is because it was frequently associated with both a higher percentage of cytokine-positive cells and greater amounts of detectable cytokines per cell . Sequential analysis of cytokine production by PB DC and monocytes after 6, 12, and 24 h of cell culture showed that after 6 h, an increased cell death rate existed among DC, which became even undetectable at 24 h, in the absence of a significant increase in cytokine secretion . In summary, our results show that from the experimental conditions assayed in this paper, to induce cytokine production by normal human DC and monocytes, maximum response is obtained once PB samples are stimulated for 6 h with LPS (with or without IFN-gamma) in the presence of brefeldin A alone . J Am Soc Echocardiogr, 2001 Mar, 14(3), 237 - 9 An unusual case of vegetative aortitis diagnosed by transesophageal echocardiography; Bansal RC et al.; We report a case of Staphylococcus aureus aortitis in a 42-year-old man who had a fever, an embolus to the left upper arm, and positive blood cultures . Transesophageal echocardiography re-vealed a 3 x 1-centimeter polypoid mass attached to the intima of the medial wall of the aorta, just distal to the origin of the left subclavian artery . The clinical presentation and the transesophageal echocardiography findings led to the diagnosis of vegetative aortitis . Antibiotic therapy was begun, and 5 days later the mass was surgically excised to prevent the possible formation of an infective aortic aneurysm and embolization to the vital organs. Biochemistry (Mosc), 2001 Jan, 66(1), 27 - 33 Thermodynamic stability and functional activity of tumor-associated antibodies; Bliznukov OP et al.; Tumor-associated antibodies of human IgG1 subclass were eluted from cell-surface antigens of human carcinoma cells and studied by differential scanning calorimetry and binding to local conformational probes, protein A from Staphylococcus aureus and a monoclonal antibody targeted to the CH2 domain of the Fc fragment . At pH 2.0-7.0, we observed virtually identical enthalpies of thermal unfolding for IgG1 from normal human sera and tumor-associated IgG1 . The exact values of calorimetric enthalpy (Delta h) at pH 7.0 were 6.1 and 6.2-6.3 cal/g for IgG1 from normal serum and IgG1 from carcinoma cells, respectively . The affinity constants of protein A binding to the CH2--CH3 domain interface demonstrated differences between serum IgG1 and tumor associated IgG1 that did not exceed 3-8-fold . The binding affinity toward the anti-CH2 monoclonal antibody determined for serum IgG1 and IgG1 from carcinoma cells differed not more than 2.5-fold . The thermodynamic parameters of IgG1 from carcinoma cells strongly suggest that protein conformational stability was essentially unaltered and that the Fc fragment of the tumor-derived IgG1 preserved its structural integrity. Vet Microbiol, 2001 Apr 2, 79(3), 267 - 74 Variation of the agr locus in Staphylococcus aureus isolates from cows with mastitis; Takeuchi S et al.; Staphylococcus aureus isolates from mastitic cow's milk were examined for production of alpha-hemolysin and protein A and their accessory gene regulator (agr locus) was analyzed . An inverse relationship between alpha-hemolysin and protein A production was found in most of the 76 isolates, suggesting that the isolates tested may be classified into group I (high alpha-hemolysin/low protein A), II (low alpha-hemolysin/high protein A), or III (low alpha-hemolysin/low protein A) . The agr locus, which consists of hld, agrB, agrD, agrC, and agrA, was detected in most of the 78 isolates including two reference strains (Wood 46 and Cowan I) by polymerase chain reaction (PCR) . When the PCR products for agr locus of 22 isolates from groups I and II were digested with restriction enzyme MboI, seven bands of the expected lengths were recognized in strain Wood 46, but not in the other isolates tested . Nucleotide sequence analysis of PCR products from six isolates revealed that the agr locus sequence of strain Wood 46 corresponded to that of the published sequence data, but the other five isolates from groups I and II diverged at agrB and agrD sequences and thus the deduced amino acid sequences . These variations of agr locus in S . aureus bovine isolates differed from those reported by Ji et al . {Science 276 (1997) 2027}. Am J Ophthalmol, 2001 Mar, 131(3), 371 - 6 Vancomycin prophylaxis and emerging resistance: are ophthalmologists the villains? The heroes? Gordon YJ. PURPOSE: To determine whether the routine use of vancomycin prophylaxis in elective cataract surgery promotes emerging resistance and provides effective protection against post-operative endophthalmitis . METHODS: Critical review of the current scientific and clinical literature was undertaken including appropriate statistical analyses of published data . RESULTS: Public health concerns for emergent vancomycin-resistant life-threatening "super bugs" are legitimate . Evaluation of the risk factors that are known to promote emerging vancomycin resistance (sick patients, hospital intensive care unit setting, methicillin-resistant Staphylococcus aureus (MRSA) clonal infections, prolonged systemic therapy, sub-therapeutic dosing, indwelling intravascular and drainage catheters, total kilogram usage and agricultural use) suggest that ophthalmic usage in routine cataract surgery is unlikely to be a significant factor in promoting emerging worldwide resistance . Clinical and scientific studies purporting to prove the value of vancomycin prophylaxis in cataract surgery contain substantial biases and design flaws that seriously undermine their validity . Issues of potential intraocular toxicity, increased costs, absence of medical-legal protection, and compliance with current Centers for Disease Control and Prevention (CDC) and American Academy of Ophthalmology guidelines (in hospital) mitigate against this practice . CONCLUSIONS: Ophthalmologists who use vancomycin prophylaxis in routine cataract surgery are neither the villains nor heroes according to my interpretation of the currently available scientific data . Personal conscience and an ongoing critical review of the literature should guide each ophthalmologist's choice in this controversy. Trends Microbiol, 2001 Mar, 9(3), 97 - 102 An embarrassment of sortases - a richness of substrates? Pallen MJ, Lam AC, Antonio M, Dunbar K. A range of surface proteins is anchored to the cell walls of Gram-positive pathogens such as Staphylococcus aureus by the transpeptidase sortase . Until now, sortase-like proteins and their substrates appeared to be limited mainly to such pathogens . However, by searching for sortase homologues among complete and incomplete genome sequences, we have found them to be present in almost all Gram-positives, a single Gram-negative bacterium and an archaean . There is usually more than one sortase-like protein encoded in each Gram-positive genome, and the genes encoding the sortase-like proteins are often clustered with genes encoding their likely substrates. Nephrol Dial Transplant, 2001 Mar, 16(3), 679 - 82; idscussion 683-5 Effect of PD fluid instillation on the peritonitis-induced influx and bacterial clearing capacity of peritoneal cells; Hekking LH et al.; BACKGROUND: The commonly used peritoneal dialysis fluids contain glucose as the osmotic agent . Heat sterilization leads to the formation of glucose degradation products which contribute, together with glucose, to the formation of advanced glycation end-products (AGEs) . AGEs have been shown to be present in the peritoneal cavity . Methods have been developed to minimize the amount of glucose degradation products in peritoneal dialysis fluids . In a rat peritoneal dialysis model, we compare the effect of a commonly used peritoneal dialysis fluid, Gambrosol, with a newly developed peritoneal dialysis fluid, PD-Bio, on the influx and functional capacity of the peritoneal cells after 2 weeks of peritoneal dialysis fluid instillation . METHODS: Three groups of animals were used: rats received daily infusion with 15 ml of either 4% Gambrosol (group 1) or 4% PD-Bio (group 2), and a control group of animals did not receive fluid (group 3) . After 2 weeks of PD fluid instillation, all the animals were injected with a 0.5 ml suspension containing 3x10(8) colony-forming units of Staphylococcus aureus . The in vivo bacterial clearing capacity was determined after 15 h . RESULTS: A statistically significant higher leukocyte influx was found in the control group compared with both PD fluid-injected groups . No statistical differences in bacterial clearing were observed among the three groups, although the number of bacteria recovered from the PD-Bio group tended to be lower than that from the Gambrosol group . Moreover, in both PD fluid instillation groups, the bacteria tended to be cleared more slowly compared with the control group . The number of mesothelial cells in the PD fluid groups was significantly greater than in the control group . CONCLUSION: No differences were observed in bacterial clearing capacity, leukocyte influx and mesothelial cell number after a 2 week exposure of the peritoneal cavity to Gambrosol vs PD-Bio. Clin Diagn Lab Immunol, 2001 Mar, 8(2), 279 - 82 Quantification of bacterial transcripts during infection using competitive reverse transcription-PCR (RT-PCR) and LightCycler RT-PCR; Goerke C et al.; Bacteria have evolved sophisticated regulatory circuits to modulate their gene expression in response to disparate environments . In order to monitor bacterial gene expression and regulation in the host, methods for direct transcript analysis from clinical specimens are needed . For most bacterial infections, amplification of the mRNAs of interest is necessary due to the low numbers of cells present and the low levels of specific transcripts . Here we compare two methods of quantitative reverse transcription-PCR (RT-PCR)-competitive RT-PCR using a one-tube system followed by standard gel analysis and the real-time detection of PCR product formation by fluorescence resonance energy transfer technology using the LightCycler unit . We isolated Staphylococcus aureus RNA directly from clinical specimens obtained from cystic fibrosis patients with chronic S . aureus lung infection and from an animal model of foreign-body infection with no further cultivation of the bacteria . Competitive RT-PCR and LightCycler RT-PCR were tested for their ability to quantify the transcription of a constitutively expressed gyrase gene (gyr) and a highly regulated alpha-toxin gene (hla) of S . aureus . Reproducible results were obtained with both methods . A sensitivity of 10(4) (gyr) and 10(3) (hla) copies, respectively, was reached, which was sufficient for the quantification of transcripts during bacterial infection . Overall, the competitive RT-PCR is a robust technique which does not need special RNA purification . On the negative side, it is labor intensive and time consuming, thus limiting the numbers of samples which can be analyzed at a given time . LightCycler RT-PCR is very susceptible to even traces of inhibitors, but it allows high-throughput processing of samples. Zh Mikrobiol Epidemiol Immunobiol, 2001 Jan-Feb, (1), 9 - 13 {Drug resistance of Staphylococcus aureus strains, isolated from children with intestinal dysbacteriosis}; Nikolaeva IV et al.; The level of antibiotic-sensitivity of 73 S . aureus strains isolated from children with dysbacteriosis of the large intestine in an outpatient clinic was determined . The isolation rate of polyresistant strains was 44% . Methicillin-resistant S . aureus (MRSA) were isolated from 25 children (34.2%) . 60% of MRSA strains could not be typed with the international set of phages . Among the strains capable of being lyzed by the phages the representatives of phage groups 3 and 4 prevailed . All MRSA strains were sensitive to vancomycin, 84-88% of the strains were sensitive to chloroamphenicol, rifampicin, spiramycin and neomycin, 80% of the strains were sensitive to fusidin and phosphomycin . The level of sensitivity of methicillin-sensitive S . aureus strains (MSSA) to different groups of antistaphylococcal antibiotics was higher . 36-64% of MRSA strains and 21-27% of MSSA strains were resistant to the action of curative bacteriophages . The suppression of obligate microflora was the risk factor in the development of staphylococcal infection of the gastrointestinal tract in children. Aust Crit Care, 2000 May, 13(2), 44 - 50 Managing central venous catheters: a prospective randomised trial of two methods; Larwood KA et al.; A randomised, prospective study was conducted to evaluate the impact on central venous catheter (CVC) infection when fluids and lines connected to a CVC were changed using a 'sterile' compared to an 'aseptic, non-touch' technique . The study sought to determine whether there were any differences in CVC tip colonisation (CTC) or CVC-related bacteraemia (CRB) as a result of the technique used for fluid and line changes . In the sterile technique (control) group, fluids and tubing were changed using full sterile technique . In the aseptic, non-touch (experimental) group, fluids and tubing attached to the CVC were changed using only a small sterile drape and a 2-minute clinical hand wash . When the CVC was removed, the tip was sampled and cultured using the semi-quantitative method . Blood cultures were also collected . In all, 111 samples from 79 patients were included in the trial: 61 in the sterile technique group and 50 in the non-touch, aseptic technique group . Results showed a CTC rate of 31 per cent in the control group and 14 per cent in the experimental group, while the CRB rate was 8.2 per cent and 6 per cent respectively . The most common organisms cultured were Staphylococcus aureus and S . epidermis respectively . This study indicates that it is safe to change fluids and lines attached to CVCs using the aseptic, non-touch technique, which has resulted in significant financial savings through less use of equipment and less nursing time required to perform fluid and line changes. Scand J Infect Dis, 2001, 33(1), 47 - 50 Primary cultures of human chondrocytes are susceptible to low inocula of Staphylococcus aureus infection and undergo apoptosis; Lee MS et al.; Hypocellularity after joint infection has been attributed to the cytotoxic effects of pus, which can cause necrosis of chondrocytes . In this study, primary cultures of human chondrocytes lost their viability and underwent necrosis rapidly with high inocula of Staphylococcus aureus infection . Chondrocytes were shown to undergo apoptosis with low inocula of Staphylococcus aureus or their culture ultrafiltrate . These findings further support the hypothesis that residual bacterial toxins or triggered apoptotic processes in chondrocytes participate in the pathogenesis of post-infectious arthropathy. J Chemother, 2001 Feb, 13(1), 34 - 42 New Staphylococcus aureus incompatibility group 1 plasmids encoding penicillinase production and resistance to different antibacterial agents; Udo EE et al.; Eleven Staphylococcus aureus plasmids encoding penicillinase production and resistance to different antibacterial agents were transferred to laboratory recipient strains in mixed-culture transfer and transduction experiments and characterized by restriction endonuclease analysis and incompatibility . The plasmids were differentiated into four types (types A-D) on the basis of their resistance phenotypes and restriction endonuclease patterns . One type encoded resistance to cadmium and arsenate . The second type encoded resistance to cadmium, mercuric compounds and nucleic acid-binding compounds . The third type encoded resistance to cadmium, kanamycin, neomycin and streptomycin while the fourth type encoded resistance to kanamycin, neomycin and ethidium bromide . Plasmids within the same class were structurally related or similar and were different from those in the other classes . Three plasmids, pWBG626, pWBG628, and pWBG663, representing three of the four plasmid types, belonged to incompatibility group 1 . These new plasmids add to the number of known incompatibility group 1 plasmids and have resistance phenotypes which should be useful for studying incompatibility of new S . aureus plasmids. J Dairy Sci, 2001 Feb, 84(2), 392 - 9 Case-control study of risk factors for clinical mastitis in postpartum dairy heifers; Waage S et al.; A case-control study was carried out to evaluate risk factors for clinical mastitis occurring in dairy heifers between 1 and 14 d postpartum . Case and control heifers were matched on herd; the control was the heifer that calved closest in time, before or after, the particular case . Data were analyzed by conditional logistic regression . The final multivariate model included 339 case-control pairs . Blood in the milk, udder edema, teat edema, and milk leakage, all recorded at the time of parturition, were significant risk factors . Purchased heifers and heifers with skin lesions between udder and thigh were not at increased risk of clinical mastitis . Separate analysis of a subgroup of case-control pairs identified teat edema, blood in the milk, and milk leakage at calving as risk factors for clinical mastitis caused by Staphylococcus aureus. Poult Sci, 2001 Feb, 80(2), 145 - 50 Systemic distribution of Staphylococcus aureus following intradermal footpad challenge of broilers; Zhu XY et al.; We conducted an experiment with broilers to determine if prior exposure to Staphylococcus aureus would facilitate the systemic infiltration of this pathogen following intradermal footpad challenge with live S . aureus . Litter-raised broilers were sensitized at 3 and 4 wk of age with s.c . injections in the neck with heat-killed S . aureus diluted in polyethylene glycol (PEG) . Equal numbers of control birds were injected at the same times with PEG . At 7 wk of age, chicks previously sensitized to killed S . aureus or injected with PEG were injected intradermally in the right footpad with PBS or live S . aureus . The left footpads of all birds were injected with PBS . The difference in thickness between the right and left footpads was determined at 0, 24, and 48 h postchallenge . Blood, liver, spleen, lung, and synovial fluid were collected six times between 1 and 48 h postchallenge to determine the recovery of S . aureus . Sensitized and non-sensitized birds showed footpad swelling following challenge with live S . aureus in the right footpad (P < 0.001) . Injection of PBS did not induce footpad swelling . Birds injected in the footpads with live S . aureus as compared to PBS had significantly higher isolation rates of S . aureus in the spleen, liver, and blood; however, recovery of S . aureus from S . aureus-sensitized and PEG-injected birds was not significantly different . Time postchallenge (1, 3, 7, 11, 24, and 48 h) had no significant effect on the recovery of S . aureus . It was concluded that the intradermal challenge of the footpad with S . aureus resulted in systemic infiltration of S . aureus into the spleen, liver, and blood . Prior exposures to killed S . aureus as compared to PEG controls did not affect the systemic distribution of S. Infect Control Hosp Epidemiol, 2001 Feb, 22(2), 99 - 104 The economic impact of methicillin-resistant Staphylococcus aureus in Canadian hospitals; Kim T et al.; OBJECTIVES: To determine the costs associated with the management of hospitalized patients with methicillin-resistant Staphylococcus aureus (MRSA), and to estimate the economic burden associated with MRSA in Canadian hospitals . DESIGN: Patient-specific costs were used to determine the attributable cost of MRSA associated with excess hospitalization and concurrent treatment . Excess hospitalization for infected patients was identified using the Appropriateness Evaluation Protocol, a criterion-based chart review process to determine the need for each day of hospitalization . Concurrent treatment costs were identified through chart review for days in isolation, antimicrobial therapy, and MRSA screening tests . The economic burden to Canadian hospitals was estimated based on 3,167,521 hospital discharges for 1996 and 1997 and an incidence of 4.12 MRSA cases per 1,000 admissions . SETtING: A tertiary-care, university-affiliated teaching hospital in Toronto, Ontario, Canada . PATIENTS: Inpatients with at least one culture yielding MRSA between April 1996 and March 1998 . RESULTS: A total of 20 patients with MRSA infections and 79 colonized patients (with 94 admissions) were identified . This represented a rate of 2.9 MRSA cases per 1,000 admissions . The mean number of additional hospital days attributable to MRSA infection was 14, with 11 admissions having at least 1 attributable day . The total attributable cost to treat MRSA infections was $287,200, or $14,360 per patient The cost for isolation and management of colonized patients was $128,095, or $1,363 per admission . Costs for MRSA screening in the hospital were $109,813 . Assuming an infection rate of 10% to 20%, we determined the costs associated with MRSA in Canadian hospitals to be $42 million to $59 million annually . CONCLUSIONS: These results indicate that there is a substantial economic burden associated with MRSA in Canadian hospitals . These costs will continue to rise if the incidence of MRSA increases further. Infect Control Hosp Epidemiol, 2001 Feb, 22(2), 105 - 8 Effectiveness of hand-cleansing agents for removing methicillin-resistant Staphylococcus aureus from contaminated hands; Guilhermetti M et al.; OBJECTIVE: The effectiveness of hand-cleansing agents in removing a hospital strain of methicillin-resistant Staphylococcus aureus from artificially contaminated hands of five volunteers was studied . DESIGN: The products used were plain liquid soap, ethyl alcohol 70% (by weight), 10% povidone-iodine liquid soap (PVP-I), and chlorhexidine gluconate (4%) detergent . The experiments were performed using a Latin square statistical design, with two 5x4 randomized blocks . The removal rates of S aureus cells from contaminated fingertips were estimated by analysis of variance, the response variable being the log10 reduction factor (RF), ie, log10 of the initial counts minus log10 of the final counts . In the first and second blocks, the fingertips of the volunteers were contaminated in mean with 3.76 log10 colony-forming units ({CFU} light-contamination hand) and 6.82 log10 CFU (heavy-contamination hand), respectively . RESULTS: In the first block, there were significant differences between treatments (P<.05) . The 10% PVP-I (RF, 3.76) and 70% ethyl alcohol (RF, 3.51) had significantly higher removal rates than plain liquid soap (RF, 1.96) and 4% chlorhexidine (RF, 1.91) . In the second block, 10% PVP-I (RF, 4.39) and 70% ethyl alcohol (RF, 3.27) also were significantly more effective than plain liquid soap (RF, 1.77) and 4% chlorhexidine (RF, 1.37; P<.05) . Plain liquid soap was significantly more effective than chlorhexidine (4%) detergent . CONCLUSIONS: The results suggest that 10% PVP-I and 70% ethyl alcohol may be the most effective hand-cleansing agents for removing methicillin-resistant S aureus strain from either lightly or heavily contaminated hands. J Chromatogr B Biomed Sci Appl, 2001 Feb 10, 751(1), 131 - 41 Sedimentation field-flow fractionation device cleaning, decontamination and sterilization procedures for cellular analysis; Battu S et al.; In Sedimentation FFF (SdFFF) practice, it is known that a large number of cell elutions create aging phenomena of the separator, thereby reducing recovery and modifying elution characteristics . Systematic cleaning procedures are developed to enhance channel lifetime, together with microbial decontamination processes . Cells can be therefore reproducibly eluted for a large number of analyses and collected under sterile conditions, if needed . This is one of the most valuable aspect if further culture or transplantation is required . Decontamination was performed using, as contaminant probe, Staphylococcus aureus, highly adherent pathogenic bacteria that eluted from SdFFF as aggregates. Vaccine, 2001 Feb 28, 19(15-16), 2092 - 9 Immunopotentiation of Staphylococcus aureus type 5 capsular polysaccharide co-entrapped in liposomes with alpha-toxin; Burgeot C et al.; Immunopotentiation of Staphylococcus aureus type 5 capsular polysaccharide (CP5) by use of liposomes as an alternative to protein-polysaccharide conjugates was investigated . Mice were immunized twice with cationic liposomes containing CP5 alone or CP5 co-entrapped with alpha-toxin or heat-detoxified alpha-toxin . Immunogenicity of these different antigens was compared with CP5-alpha-toxin conjugates . Antibodies against CP5 were elicited in mice immunized with conjugates or liposomes containing co-entrapped CP5 and alpha-toxin . Liposomes containing CP5 alone or co-entrapped CP5 and alpha-toxoid failed to induce antibodies against CP5 . All the preparations entailed an antibody response against alpha-toxin and highest antibody and neutralizing activity titers were obtained with liposomes . These results show that liposomes can be used to immunopotentiate CP5, however, a co-incorporated protein able to insert lipids bilayers is probably required. J Endocrinol Invest, 2001 Jan, 24(1), 45 - 50 Pituitary abscess presenting with cranial nerve paresis . Case report and review of literature; Somali MH et al.; Non-adenomatosus lesions of the pituitary represent a small part of the intrasellar processes and they have heterogeneous presentation . Making a precise diagnosis is of great importance, as it may lead to more efficient management . A 65-year-old man was admitted to the hospital because of headache and right cranial nerve III palsy . Basic laboratory work-up was normal whereas endocrinological assessment revealed hypopituitarism without diabetes insipidus . Plain radiography showed an enlarged sella and frontal and paranasal sinusitis . Computed tomography (CT) and magnetic resonance imaging (MRI) of the sella revealed an intrasellar lesion with extension to the sphenoid and cavernous sinuses as well as the suprasellar region, exerting pressure on the optic chiasm . On T1-weighted images the mass had a low-intensity signal with a smooth enhancing rim with bright signal . Given the presence of multiple sinusitis and imaging characteristics a pre-operative diagnosis of pituitary abscess was made . The patient was operated via transphenoidal route and purulent material was drained out . Cultures of the material were positive for Staphylococcus aureus . Antibiotics as well as cortisol replacement therapy were given . Three months later hypopituitarism persisted but there was significant improvement in the neurological findings . We report a case of an unusual presentation of a pituitary abscess . High index of suspicion, the presence of associated conditions such as pituitary tumors, meningitis or sinusitis, as well as diabetes insipidus and specific imaging features are the main diagnostic clues . Pre-operative diagnosis, which will lead to prompt antibiotic therapy and transphenoidal drainage, can decrease high mortality and morbidity associated with this disease. Vutr Boles, 2000, 32(2), 35 - 40 {Activity of vancomycin and teicoplanin against clinical isolates of Staphylococcus aureus in the period 1994-1999}; Nashev D et al.; Since the emergence of the methicillin-resistant S . aureus (MRSA) in the 1960's, glycopeptides (Vancomycin and Teicoplanin) has been the drugs of choice and commonly the sole antimicrobial agents available for the treatment of serious MRSA and other Gram-positive infections . The emergence of S . aureus with intermediate vancomycin-resistance after 1997 threatens to return us to the era before the development of the antibiotics . Prevention of the further spread of S . aureus strains with intermediate and eventually with full glycopeptide resistance requires enhanced laboratory methods to detect resistance . A total of 361 S . aureus clinical isolates (177 MRSA and 184 MSSA) obtained from 1994 to 1999 in eleven Bulgarian hospitals located in geographically distinct areas of the country were enrolled in the study . Minimal inhibitory concentrations of Vancomycin and Teicoplanin were determined by agar-dilution method according to NCCLS recommendations . MIC50 and MIC90 for Vancomycin were 0.7 and 1 mg/ml, and for Teicoplanin--0.5 and 0.9 microgram/ml . All staphylococcal isolates showed sensitivity to Vancomycin and Teicoplanin . MICs of both glicopeptides against MRSA and MSSA did not differ significantly. Int J Parasitol, 2001 Mar, 31(3), 265 - 71 Down-regulation of human B lymphocyte activities by a Trypanosoma cruzi membrane glycoprotein; Kierszenbaum F et al.; The effects of purified AGC10, a Trypanosoma cruzi membrane glycoprotein, on normal human B lymphocytes were studied in this work . In the presence of AGC10, {3H}-thymidine uptake by human peripheral blood mononuclear cells stimulated with the B cell-specific mitogen SACI (killed Staphylococcus aureus Cowan I) was markedly decreased . This alteration was accompanied by others such as decreased expression of the CD122 and CD132 chains of the IL-2R complex . These inhibitory effects appeared to be somewhat selective, as expression of CD25, another IL-2R chain, was not affected by AGC10 and no significant modification occurred in the expression of the B-cell-specific marker CD19 or CD21 . In contrast, AGC10 did reduce the levels of expression of CD86 and CD80, molecules known to play critical roles in B cell interactions with T lymphocytes . Fairly large subpopulations of, but not all, B lymphocytes had their expression of CD122(+), CD132(+), CD86(+) and CD80(+) reduced to undetectable levels in the presence of AGC10 . However, the SACI-activated B cells that remained capable of expressing these molecules in the presence of AGC10 did so at normal levels . This was denoted by comparable mean fluorescence intensity values representing the expression of CD122, CD132, CD86 or CD80 molecules on the surface of SACI-stimulated CD19(+) cells cultured without or with AGC10 . These results indicated that AGC10, derived from an organism that causes immunosuppression in infected hosts, down-regulates B cell activities and suggested that the relevant mechanism could involve the molecular alterations described above. Nippon Rinsho, 2000 Nov, 58(11), 2338 - 43 {Toxic shock syndrome complicating influenza}; Nagasaka Y; In January 1999, an outbreak of type A(H3N2) influenza occurred in a psychiatric hospital . Among 273 inpatients, 59 male and 40 female patients, age between 24-84 year old, developed high fever(38 degrees C), joint pain, cough and nasal discharge, and eleven male and one female patients died . In that hospital, Methicillin-resistant Staphylococcus aureus(MRSA), with production of toxic shock syndrome toxin-1 (TSST-1) and staphylococcal enterotoxin(SE) type C(SEC), were isolated from inpatients, medical stuffs and hospital environments . Under investigation, a 59-year-old male inpatient developed respiratory failure following influenza infection . The titer of serum TSST-1 antibody and SEC antibody of this patient was significantly elevated in his clinical course . It is highly suspected that influenza outbreak occurred among MRSA carriers and TSS was complicated with fatal cases. Nat Immunol, 2001 Mar, 2(3), 223 - 8 Specific missense mutations in NEMO result in hyper-IgM syndrome with hypohydrotic ectodermal dysplasia; Jain A et al.; The gene that encodes nuclear factor kappaB (NF-kappaB) essential modulator (or NEMO, also known as IKKgamma) is required for activation of the transcription factor NF-kappaB . We describe mutations in the putative zinc-finger domain of NEMO that result in an X-linked primary immunodeficiency characterized by hyper-IgM syndrome and hypohydrotic ectodermal dysplasia (XHM-ED).These mutations prevent CD40 ligand (CD40L)-mediated degradation of inhibitor of NF-kappaB alpha (IkappaB-alpha) and account for the following observations: B cells from XHM-ED patients are unable to undergo immunoglobulin class-switch recombination and antigen-presenting cells (APCs) are unable to synthesize the NF-kappaB-regulated cytokines interleukin 12 (IL-12) or tumor necrosis factor alpha (TNF-alpha) when stimulated with CD40L . Nevertheless, innate immunity is preserved in XHM-ED patients because APCs retain the capacity to respond to stimulation by lipopolysaccharide or Staphylococcus aureus Cowan's antigen (SAC) . Overall, the phenotype observed in XHM-ED patients shows that the putative zinc-finger domain of NEMO has a regulatory function and demonstrates the definite requirement of CD40-mediated NF-kappaB activation for B cell immunoglobulin class-switching. J Bacteriol, 2001 Mar, 183(6), 1843 - 52 Sigma(B) activity depends on RsbU in Staphylococcus aureus; Giachino P et al.; Derivatives of the widely used laboratory strain Staphylococcus aureus NCTC8325, which are natural rsbU mutants, were shown to be unable to produce RsbU, a positive regulator of the alternative sigma factor sigma(B) . The lack of RsbU prevented the heat-dependent production of sigma(B)-controlled transcripts and resulted in reduced H2O2 and UV tolerance, enhanced alpha-hemolysin activity, and the inability to produce the alkaline shock protein Asp23 . After 48 h of growth, rsbU mutant strains failed to accumulate staphyloxanthin, the major stationary-phase carotenoid . Transcription of Asp23 was found to be exclusively controlled by sigma(B), making it an excellent target for the study of sigma(B) activity in S . aureus . Reporter gene experiments, using the firefly luciferase gene (luc+) fused to the sigma(B)-dependent promoter(s) of asp23, revealed that sigma(B) is almost inactive in 8325 derivatives . cis complementation of the 8325 derivative BB255 with the wild-type rsbU gene from strain COL produced the rsbU(+) derivative GP268, a strain possessing a sigma(B) activity profile comparable to that of the rsbU(+) wild-type strain Newman . In GP268, the heat inducibility of sigma(B)-dependent genes, Asp23 production, alpha-hemolysin activity, pigmentation, and susceptibility to H2O2 were restored to the levels observed in strain Newman, clearly demonstrating that RsbU is needed for activation of sigma(B) in S . aureus. J Antimicrob Chemother, 2001 Mar, 47(3), 349 - 52 Efficacy of linezolid in a staphylococcal endocarditis rabbit model; Oramas-Shirey MP et al.; A rabbit endocarditis model was used to test the efficacy of oral linezolid and iv vancomycin . Twenty-four hours after catheter placement across the aortic valve, rabbits were infected with 3.5 x 10(6) cfu of Staphylococcus aureus (UC-9258) . Two days after infection, control rabbits were killed, and treated rabbits were given 5 days of therapy with linezolid at 8 h intervals (tds) using either 25, 50 or 75 mg/kg/dose, or vancomycin at 12 h intervals (bd) using 25 mg/kg/dose . Linezolid at 75 and 50 mg/kg, and vancomycin significantly reduced S . aureus in aortic valve vegetations compared with the control . Linezolid at 25 mg/kg was ineffective . The efficacy of 75 and 50 mg/kg linezolid was related to maintenance of plasma drug levels near or above the linezolid MIC for UC-9258 (2 mg/L). J Antimicrob Chemother, 2001 Mar, 47(3), 277 - 83 Fitness and competitive growth advantage of new gentamicin-susceptible MRSA clones spreading in French hospitals; Laurent F et al.; Since 1991, new epidemic methicillin-resistant Staphylococcus aureus (MRSA) strains characterized by the unexpected reappearance of heterogeneous phenotypic expression of resistance to methicillin and by susceptibility to gentamicin and various other antibiotics (GS-MRSA) have been reported in France . GS-MRSA strains have progressively replaced MRSA clones expressing homogeneous resistance to methicillin and resistance to gentamicin (GR-MRSA) . In this study, we investigated the physiological characteristics of these new clones . In particular, we evaluated and compared the maximal growth rate and the deduced generation times (related to fitness of strains) of the major French epidemic MRSA clones . The population studied consisted of 79 isolates including (i) GR-MRSA that comprised six different types on the basis of PFGE; (ii) GS-MRSA the majority of which clustered into two PFGE types, A1 (usually resistant to erythromycin) and B (usually susceptible to erythromycin); (iii) methicillin-susceptible S . aureus (MSSA) . GS-MRSA-A1 and MSSA strains were shown to have a significant fitness benefit (about 20%) with shorter generation times (theta = 23.7 +/- 0.1 and 22.9 +/- 0.05 min, respectively) than GR-MRSA and GS-MRSA-B strains (theta = 30.3 +/- 0.2 and 32.5 +/- 0.5 min, respectively) . These data suggest that a link exists between genetic patterns, resistance profiles and physiological properties . In vitro competitive experiments indicated that GS-MRSA- A1 strains were able to rapidly outgrow GR-MRSA strains . The growth advantage observed should be taken into account in understanding the spread of some new clones of MRSA. J Antimicrob Chemother, 2001 Mar, 47(3), 261 - 70 Effect of hydrophobicity and molecular mass on the accumulation of fluoroquinolones by Staphylococcus aureus; Piddock LJ et al.; Ten novel fluoroquinolones, with similar chemical structures but differing antibacterial activities and hydrophobicities, were studied to evaluate the role of the physical properties of quinolones on their accumulation and antibacterial activity for Staphylococcus aureus . Six of the 10 agents and tosufloxacin were more active against quinolone-susceptible and -resistant S . aureus than the remaining four agents and several piperazinyl fluoroquinolones . Changes to the side chains of the pyrollidinyl substituent at the R7 position alone made little difference to the MICs . Comparison of MICs of agents that were structurally identical apart from the R1 substituents, confirmed that a t-butyl group confers enhanced activity against S . aureus over a cyclopropyl or ethyl group at this position . The steady-state concentrations (SSCs) of the 10 novel quinolones accumulated by wild-type S . aureus did not correlate with their MICs or chemical structures . There was no apparent relationship between logP of the quinolone and accumulation by S . aureus F77; however, accumulation was positively correlated with molecular mass for 9/10 agents (r = 0.745) confirming that high molecular mass is not a barrier to accumulation in S . aureus . For all 10 agents, the presence of carbonyl cyanide m-chlorophenylhydrazone (CCCP) increased the concentration of quinolone accumulated by SA-1199, suggesting that NorA was inhibited . The fold increase of the SSC in the presence of CCCP did not correlate with hydrophobicity, but the SSC of agents with either an ethyl or cyclopropyl group at R1 was increased two- to three-fold in the presence of CCCP, suggesting that affinity for the NorA efflux pump may be influenced by quinolone structure. Int J Clin Pract, 2001 Jan-Feb, 55(1), 59 - 63 Linezolid; French G; Multiple antibiotic resistance is increasing worldwide in Gram-positive bacteria, especially in hospitals . Problem organisms include multiply resistant strains of pneumococci, Staphylococcus aureus and enterococci; for many of these the glycopeptide vancomycin has become the treatment of last resort . This situation has been made worse by the emergence of vancomycin-resistant enterococci and vancomycin-intermediate S . aureus . Fortunately, several compounds active against resistant Gram-positive bacteria are in active development . One of these is linezolid, the first of the oxazolidinones, a new class of antibacterial . Linezolid is a synthetic agent which is active against all the clinically important Gram-positive bacteria, including multiply resistant strains . It has good pharmacokinetics, with equal bioavailability by both oral and intravenous routes and no need for dose adjustment in patients with renal impairment . The drug has a good safety profile and clinical trials have given excellent results in a variety of skin and soft tissue, respiratory and bloodstream infections . Linezolid is a promising drug, which, together with prudent antibiotic use and the prevention and control of hospital infection, will help in the battle against multiply antibiotic resistant Gram-positive bacteria. Int J Clin Pract Suppl, 2000 Dec, (115), 44 - 9 Staphylococcus aureus small colony variants: formation and clinical impact; von Eiff C et al.; S . aureus small-colony variants are a naturally occurring subpopulation which grow slowly and produce small colonies on routine media . They also demonstrate a number of other characteristics that are atypical for S . aureus including reduced alpha-toxin production and delayed coagulase activity . The connection of S . aureus SCVs with persistent and relapsing infections has been defined over the past decade, especially in patients with chronic osteomyelitis and in cystic fibrosis patients as demonstrated by prospective studies . While the studies with clinical isolates of SCVs suggested a link between electron transport-defective strains and persistent infections, a defined hemB mutant with the SCV phenotype provided strong additional evidence for these connections . The hemB mutant was phagocytosed by cultured endothelial cells, but did not lyse these cells, because the mutant produced very little alpha-toxin . The intracellular location may shield the SCVs from host defenses and antibiotics, thus providing one explanation for the difficulty in clearing S . aureus SCVs from host tissues. Swiss Med Wkly, 2001 Jan 27, 131(3-4), 35 - 40 CD14 expression on monocytes and TNF alpha production in patients with septic shock, cardiogenic shock or bacterial pneumonia; de Werra I et al.; OBJECTIVES: In patients with septic shock, circulating monocytes become refractory to stimulation with microbial products . Whether this hyporesponsive state is induced by infection or is related to shock is unknown . To address this question, we measured TNF alpha production by monocytes or by whole blood obtained from healthy volunteers (controls), from patients with septic shock, from patients with severe infection (bacterial pneumonia) without shock, and from patients with cardiogenic shock without infection . MEASUREMENTS: The numbers of circulating monocytes, of CD14+ monocytes, and the expression of monocyte CD14 and the LPS receptor, were assessed by flow cytometry . Monocytes or whole blood were stimulated with lipopolysaccharide endotoxin (LPS), heat-killed Escherichia coli or Staphylococcus aureus, and TNF alpha production was measured by bioassay . RESULTS: The number of circulating monocytes, of CD14+ monocytes, and the monocyte CD14 expression were significantly lower in patients with septic shock than in controls, in patients with bacterial pneumonia or in those with cardiogenic shock (p < 0.001) . Monocytes or whole blood of patients with septic shock exhibited a profound deficiency of TNF alpha production in response to all stimuli (p < 0.05 compared to controls) . Whole blood of patients with cardiogenic shock also exhibited this defect (p < 0.05 compared to controls), although to a lesser extent, despite normal monocyte counts and normal CD14 expression . CONCLUSIONS: Unlike patients with bacterial pneumonia, patients with septic or cardiogenic shock display profoundly defective TNF alpha production in response to a broad range of infectious stimuli . Thus, down-regulation of cytokine production appears to occur in patients with systemic, but not localised, albeit severe, infections and also in patients with non-infectious circulatory failure . Whilst depletion of monocytes and reduced monocyte CD14 expression are likely to be critical components of the hyporesponsiveness observed in patients with septic shock, other as yet unidentified factors are at work in this group and in patients with cardiogenic shock. Kansenshogaku Zasshi, 2001 Jan, 75(1), 14 - 9 {Isolation of methicillin-resistant Staphylococcus aureus from nasopharyngeal swabs on admission to a ward for pediatric patients: comparison between 1992-1993 and 1997-1998}; Sakata H; We studied 2,176 nasopharyngeal swabs obtained from 1,891 children on admission to the pediatric ward at Asahikawa Kosei Hospital from January 1997 to December 1998, and compared the results with the same study from January 1992 to December 1993 . Fifty strains of Methicillin-resistant Staphylococcus aureus (MRSA) were isolated from 46 patients . The ages of these patients ranged from 7 days to 17 years and 24 patients were younger than 1 year . Six patients were diagnosed as MRSA infection; acute otitis media in 3, osteomyelitis and bacteremia in 1, staphylococcal scalded skin syndrome in 1, and inflammation in umbilical region in 1 . The ratio of MRSA positive to the total specimens significantly increased from 1.25% in 1992-1993 to 2.30% in 1997-1998 (p = 0.011) . The ratio of MRSA positive to the total S . aureus positive specimens significantly increased from 10.7% in 1992-1993 to 20.3% in 1997-1998 (p = 0.003) . These findings demonstrated that prevalence of MRSA carriers among children is increasing. J Antibiot (Tokyo), 2000 Dec, 53(12), 1346 - 53 A potent seryl tRNA synthetase inhibitor SB-217452 isolated from a Streptomyces species; Stefanska AL et al.; A potent inhibitor of seryl tRNA synthetase, designated SB-217452 has been isolated from Streptomyces sp . ATCC 700974 . The fermentation, isolation, structure elucidation and some properties are described . SB-217452 showed inhibitory activity against both Staphylococcus aureus and rat seryl tRNA synthetases, with similar IC50 values of approximately 8 nM . The inhibitor is the serine linked nucleoside moiety of the antibiotic albomycin delta2 . In contrast to albomycin delta2, SB-217452 showed only very weak antibacterial activity against a limited range of microorganisms . The compound has not been previously reported as a naturally occurring metabolite . In addition to SB-217452, albomycin delta2 Fe3+ complex and the novel Al3+ complex were isolated from the fermentation . These complexes had no seryl tRNA synthetase inhibitory activity. Perit Dial Int, 2000 Nov-Dec, 20(6), 625 - 30 Prevention of peritonitis in children on peritoneal dialysis; Verrina E et al.; We reviewed methods of preventing peritonitis in children . A considerable body of evidence indicates that peritonitis rates are lowest with the use of a double-cuffed catheter, with a downward directed tunnel, placed by an experienced surgeon . Evidence in adults, but lacking in children, suggests that exit-site mupirocin will lower Staphylococcus aureus exit-site infections and thus peritonitis rates . The risk of peritonitis due to contamination can be diminished by the avoidance of spiking and by the provision of a long training period . Catheter removal and replacement for catheter-related peritonitis may be done simultaneously in certain circumstances and is useful in decreasing the risk of recurrent peritonitis . Antibiotic prophylaxis at the time of catheter insertion, for contamination, during dialysate leaks, and for invasive procedures appears to be useful in diminishing peritonitis risk. Basic Res Cardiol, 2001 Feb, 96(1), 75 - 81 Antiendothelial antibodies in sera of patients with infective endocarditis; Portig I et al.; Infective endocarditis is characterized by the colonization of endocardium by microorganisms . Except for Staphylococcus aureus, microorganisms are not able to adhere to and grow on endocardial cells; prior damage, e.g., by shear stress or other mechanical factors, is necessary . But other causes may well have a share . This study was, therefore, designed to identify immunological factors, especially antibodies against endothelial cells, which could contribute to the initiation of endocardial injury . Sera of patients with infective endocarditis and healthy controls were investigated for the presence of antibodies against endothelial antigen . As the antigen source human umbilical vein endothelial cells were used . Antibodies against endothelial cells were detected by indirect immunofluorescence, ELISA, immunoblotting, antibody dependent cellular cytotoxicity, and antibody mediated cytotoxicity . Antibodies against endothelial cells were found in seven out of fifteen patients . These antibodies were directed against cytoplasmic structures and only appeared in the course of the disease . A correlation between the presence of these antibodies and disease activity or the outcome of disease was not observed . These antibodies may develop as a consequence of damage to endocardial cells (thereby exposing intracellular antigen to the immune system) and do not seem to play a role in the pathogenesis of infective endocarditis. J Med Liban, 2000 Jul-Aug, 48(4), 203 - 7 Molecular diagnosis of antimicrobial resistance; Ahmad S et al.; Advances in molecular technology have helped in better understanding of mechanisms and diagnosis of diseases in many medical fields . Several molecular techniques are available for determining the genotypic drug-resistance and monitoring epidemic spread of a particular antimicrobial resistance gene in a hospital or patient population . The molecular (genotypic) testing has several advantages over conventional (phenotypic) testing in being faster and unambiguous, more accurate, able to detect masked resistance and can serve as a "gold" or "reference" test for detecting antibiotic resistance genes . This article addresses these molecular tests with their application and limitations and provide examples of their use especially in Mycobacterium tuberculosis and methicillin resistant Staphylococcus aureus. Lancet, 2001 Jan 27, 357(9252), 299 - 301 The fourth disease, 1900-2000; Weisse ME; Measles and scarlet fever were differentiated from one another in the 17th century . Rubella was accepted as the third distinct paediatric exanthem in 1881 . Nil Filatow in 1885 and Clement Dukes in 1894 described two distinct forms of rubella, and in 1900 Dukes proposed that one of these forms of rubella was a separate entity which he called the fourth disease . For the past five decades, fourth disease has been considered a non-entity, perhaps a mild form of scarlet fever, but certainly not a distinct disease . In 1979 Keith Powell resurrected the idea of the fourth disease and argued that it was caused by exotoxin-producing Staphylococcus aureus . We present additional arguments for the existence of fourth disease, as well as information to link the disease to S . aureus. Biosci Biotechnol Biochem, 2000 Dec, 64(12), 2631 - 43 Prophage, phiPV83-pro, carrying panton-valentine leukocidin genes, on the Staphylococcus aureus P83 chromosome: comparative analysis of the genome structures of phiPV83-pro, phiPVL, phi11, and other phages; Zou D et al.; Staphylococcus aureus P83 has Panton-Valentine leukocidin (PVL)-like genes, lukM and lukF-PV . Here, lukM and lukF-PV genes were found on the genome of a prophage, which was designated as phiPV83-pro . The precise genome size was 45,636 bp with att core sequences of 10 base pairs . Sixty-four ORFs were identified on the phiPV83-pro genome, including two extra operons, lukM-lukF-PV and orfs63-64 . The lukM-lukF-PV cluster was located 2.1 kb upstream of the attL site . The most striking feature of the phiPV83-pro genome was a constituent of at least 4 regions from phi11, phiPVL, and other phages, i.e., (i) att sites identical with those of phi11, (ii) a cos sequence and the genes encoding packaging and head proteins of phiPVL (occupied half region of phiPV83-pro), and (iii) the other two regions which showed no significant similarity with known phages (occupied about 40% of phiPV83-pro) . Furthermore, two insertion sequences, ISSA1 and ISSA2 were integrated into attL site and orf44, respectively . PhiPV83-pro was not induced as phage particles from S . aureus P83 regardless of its treatment with mitomycin C . The insertion of ISSA1 into the attL site was one of the reasons of the failure of the induction of the phage particles by mitomycin C treatment of the strain P83. New Microbiol, 2001 Jan, 24(1), 57 - 61 Adhesion of a Staphylococcus aureus strain to biomaterials does not select methicillin-resistant mutants; Montanaro L et al.; Bacterial adhesion to polymethylmethacrylate and to silicon elastomer, materials frequently used in clinical applications, has been investigated to assess whether adhesion selects methicillin-resistant mutants in the bacterial population in contact with the materials . The methicillin susceptibility of a susceptible Staphylococcus aureus (ATCC 25923) was measured by a modification of plate antibiogram Kirby-Bauer method, which allows optimised detection of small variations in antibiotic susceptibility . In both adherent and non-adherent bacterial subpopulations, the presence of mecA gene, which encodes for the protein PBP 2a responsible for methicillin resistance was searched for by Polymerase Chain Reaction (PCR) . The contact with the two polymers did not induce in the bacteria population any phenotypic increase in methicillin resistance, or the selection of mutants carrying the mecA gene. Eur J Clin Microbiol Infect Dis, 2000 Dec, 19(12), 953 - 5 Characterization of epidemic and nonepidemic methicillin-resistant Staphylococcus aureus strains in a university hospital in northeast Germany; Panzig B et al.; The aim of this study was to analyze methicillin-resistant Staphylococcus aureus strains isolated during a 1-year period by means of pulsed-field gel electrophoresis, resistance phenotyping and determination of biochemical features . Eight different resistance phenotypes with the predominant resistance type Pen Oxa Cip (penicillin, oxacillin, ciprofloxacin) were observed . None of the strains tested exhibited decreased susceptibility to vancomycin, but two strains were resistant to mupirocin . Genetic relatedness of methicillin-resistant Staphylococcus aureus isolates could be shown for two outbreaks, one of which was caused by a clone with an epidemic potential concerning duration of colonization/infection of patients and dissemination of the strains in the hospital. Br J Biomed Sci, 2000, 57(4), 269 - 72 Methicillin-resistant Staphyloccocus aureus: evaluation of five selective media; Davies S et al.; This study evaluates the performance in isolating methicillin-resistant Staphylococcus aureus (MRSA) of three media: the reduced salt formulation of mannitol salt agar plus oxacillin (MMSAO); CHROMagar Staph aureus plus ciprofloxacin (CHRAC); and Halifax MRSA medium (HMO), against the previously recommended mannitol salt agar (7% salt) plus oxacillin (OMSAO) and Baird-Parker medium plus ciprofloxacin (BPC) . MRSA screening swabs were plated out onto the five selective media and the plates examined at 24 and 48 h . Suspected colonies were confirmed as MRSA by detection of heat-labile DNase, coagulase and/or protein A, and by confirming resistance to methicillin . Of 719 specimens examined, 191 grew MRSA on at least one medium . The relative sensitivities of the five media at 48 h were as follows: BPC, 94%; CHRAC, 70%; OMSAO, 61%; HMO, 56%; and MMSAO, 46% . In addition, BPC gave the least number of unnecessary investigations for non-MRSA isolates . The current advantage of BPC when performing direct culture for MRSA was confirmed . The other ciprofloxacin-containing medium also produced reasonable results . Of the two mannitol salt agar media, the formulation with 7% salt gave better results . HMO proved unreliable at isolating MRSA. Scand J Infect Dis, 2000, 32(6), 587 - 95 Spread of Staphylococcus aureus in hospitals: causes and prevention; Solberg CO; Methicillin-resistant Staphylococcus aureus (MRSA) has become a major nosocomial pathogen in many hospitals worldwide . Even more alarming, MRSA strains that are vancomycin intermediate-susceptible are isolated with increasing frequency, making therapy for staphylococcal infections even more difficult and prevention more important than ever . Spread of S . aureus in hospitals and infection control measures are reviewed . The major sources of S . aureus in hospitals are septic lesions and carriage sites of patients and personnel . Carriage often precedes infection . The anterior nares are the most consistent carriage site, followed by the perineal area . Skin contamination and aerial dissemination vary markedly between carriers and are most pronounced for combined nasal and perineal carriers . The principal mode of transmission is via transiently contaminated hands of hospital personnel . Airborne transmission seems important in the acquisition of nasal carriage . Infection control strategies include screening and isolation of newly admitted patients suspected of carrying MRSA or S . aureus with intermediate resistance to vancomycin, implementation of an infection control program to prevent transmission of resistant strains between patients and hospital personnel, and institution of a proper antibiotic policy to minimize antibiotic resistance development . MRSA carriers should be treated with intranasal antibiotics, e.g . mupirocin, and skin disinfectants to eliminate carriage . Education of hospital personnel is essential . Improved knowledge about the best ways to ensure favourable infection control practices is needed . Active intervention against the spread of MRSA is important. J Food Prot, 2001 Jan, 64(1), 51 - 7 Modeling the growth boundary of Staphylococcus aureus for risk assessment purposes; Stewart CM et al.; Knowing the precise boundary for growth of Staphylococcus aureus is critical for food safety risk assessment, especially in the formulation of safe, shelf-stable foods with intermediate relative humidity (RH) values . To date, most studies and resulting models have led to the presumption that S . aureus is osmotolerant . However, most studies and resulting models have focused on growth kinetics using NaCl as the humectant . In this study, glycerol was used to investigate the effects of a glass-forming nonionic humectant to avoid specific metabolic aspects of membrane ion transport . The experiments were designed to produce a growth boundary model as a tool for risk assessment . The statistical effects and interactions of RH (84 to 95% adjusted by glycerol), initial pH (4.5 to 7.0 adjusted by HC1), and potassium sorbate (0, 500, or 1,000 ppm) or calcium propionate (0, 500, or 1,000 ppm) on the aerobic growth of a five-strain S . aureus cocktail in brain heart infusion broth were explored . Inoculated broths were distributed into microtiter plates and incubated at 37 degrees C over appropriate saturated salt slurries to maintain RH . Growth was monitored by turbidity during a 24-week period . Toxin production was explored by enterotoxin assay . The 1,280 generated data points were analyzed by SAS LIFEREG procedures, which showed all studied parameters significantly affected the growth responses of S . aureus with interactions between RH and pH . The resulting growth/no growth boundary is presented. J Pharm Pharmacol, 2000 Dec, 52(12), 1547 - 52 Electron-microscopic study of the bactericidal effect of OPB-2045, a new disinfectant produced from biguanide group compounds, against methicillin-resistant Staphylococcus aureus; Sakagami Y et al.; The bactericidal effect of OPB-2045, a new disinfectant produced from biguanide group compounds, against methicillin-resistant Staphylococcus aureus (MRSA), MRSA IID 1677, was investigated by transmission electron microscopy . OPB-2045 showed strong bactericidal activity against MRSA . The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of OPB-2045 against the test strain were 0.78 and 1.56 microg mL(-1), respectively . The test bacteria were incubated in the presence of OPB-2045 at 1/2 MIC (0.39 microg mL(-1)), 1 MIC (0.78 microg mL(-1)), 2 MIC (1 MBC, 1.56 microg mL(-1)), 4 MIC (2 MBC, 3.13 microg mL(-1)) or 10 MIC (5 MBC, 7.8 microg mL(-1)) at 37 degrees C for 30 s, 3 min, 30 min or 6h . The morphology of the cells was examined by transmission electron microscopy . The cell damage observed after 30-min or 6-h incubation in the presence of OPB-2045 at 1/2 or 1 MIC was the same as that at 2, 4 or 10 MIC . The numbers of damaged MRSA cells increased according to the increase in concentration of added disinfectant, and the image of bacteriolysis was observed, too . After treatment at 1/2 or 1 MIC, a few leaking cells were recognized, but no destroyed cells were found . No morphological changes were observed after treatment at 1 or 2 MIC for 30 s, 3 min or 30 min . When the incubation time was extended to 6 h, morphological changes in the MRSA cells treated at 1 or 2 MIC were observed . When examining the relationship between the numbers of surviving bacteria and the MIC (MBC) values in soybean casein digest broth, no decrease in MRSA cell numbers was recognized in the untreated control or at 1/2 MIC, but a marked decrease in MRSA cell numbers was recognized as the OPB-2045 concentration was increased . The new disinfectant OPB-2045 would make a useful contribution to the medical field for the prevention of infections caused by pathogenic bacteria such as MRSA. Nature, 2001 Jan 11, 409(6817), 215 - 9 Crystal structures of SarA, a pleiotropic regulator of virulence genes in S . aureus; Schumacher MA et al.; Staphylococcus aureus is a major human pathogen, the potency of which can be attributed to the regulated expression of an impressive array of virulence determinants . A key pleiotropic transcriptional regulator of these virulence factors is SarA, which is encoded by the sar (staphylococcal accessory regulator) locus . SarA was characterized initially as an activator of a second virulence regulatory locus, agr, through its interaction with a series of heptad repeats (AGTTAAG) within the agr promoter . Subsequent DNA-binding studies have revealed that SarA binds readily to multiple AT-rich sequences of variable lengths . Here we describe the crystal structure of SarA and a SarA-DNA complex at resolutions of 2.50 A and 2.95 A, respectively . SarA has a fold consisting of a four-helix core region and 'inducible regions' comprising a beta-hairpin and a carboxy-terminal loop . On binding DNA, the inducible regions undergo marked conformational changes, becoming part of extended and distorted alpha-helices, which encase the DNA . SarA recognizes an AT-rich site in which the DNA is highly overwound and adopts a D-DNA-like conformation by indirect readout . These structures thus provide insight into SarA-mediated transcription regulation. Poult Sci, 2000 Dec, 79(12), 1851 - 6 Shelf life of ground poultry meat stored under modified atmosphere; Saucier L et al.; The shelf life of ground chicken and turkey meat packaged under a modified atmosphere containing O2 and a high level of CO2 (62% CO2, 8% O2, and 30% N2; gas-1), or a gas mixture without O2 (20% CO2 and 80% N2; gas-2) was evaluated for 20 d at 1 C . Meat packaged under gas-2 maintained a higher a* value (redness) throughout the experiment and generally had a more appealing color than the meat packaged using gas-1 . Microbial populations were assessed after 8, 12, and 15 d of storage . Total aerobic mesophilic counts were higher in chicken meat than in turkey throughout storage . Coliforms and Escherichia coli counts were lower in meat packaged under gas-1 . After 15 d of storage at 1 C, Brochothrix thermosphacta was detected only in ground chicken meat packaged using gas-2 . Meat packaged under the gas mixtures tested had similar counts for presumptive pseudomonads, Staphylococcus aureus, and lactic acid bacteria . These results indicate that an appropriate gas mixture can maintain a desirable color in ground poultry meat but offers no guarantees with respect to the microbial profile of meat. J Med Microbiol, 2001 Jan, 50(1), 35 - 41 Involvement of staphylococcal protein A and cytoskeletal actin in Staphylococcus aureus invasion of cultured human oral epithelial cells; Jung KY et al.; Following the coincidental discovery that beta-actin isolated from renal epithelial cells was precipitated by staphylococcal protein A (SPA), the possibility that SPA and cytoskeletal actin filaments may be involved in Staphylococcus aureus infection of epithelial cells was considered . Therefore, to clarify the potential role of SPA and actin filaments in S . aureus infection, the invasion efficiency of S . aureus was determined quantitatively by measuring the number of cfu of viable organisms recovered from cultured KB cells . S . aureus invasion was found to be time dependent (0-60 min) and increased linearly when increasing numbers of bacteria were added (10(4)-10(6) cfu/ml) . However, significant variation in the level of invasion was noted in protein A-deficient S . aureus Wood 46 . Cytochalasin B inhibited the invasion efficiency of S . aureus in a dose-dependent manner . The present study suggests that interaction of staphylococcal protein A and cytoskeletal actin filaments is involved in the S . aureus invasion of cultured KB cells, and this process may contribute, in part, to the intracellular movement, cell-to-cell spread and dissemination of S . aureus within human oral epithelial cells in vivo. J Med Microbiol, 2001 Jan, 50(1), 104 - 6 Increases in the mutation frequency at which fusidic acid-resistant Staphylococcus aureus arise with salicylate; Price CT et al.; Salicylate was shown to increase the frequency at which a fusidic acid-susceptible strain of Staphylococcus aureus underwent mutation to become fusidic acid-resistant . These fusidic acid-resistant mutants had alterations in spectinomycin and kanamycin resistance levels indicative of mutations in fusA, the gene that encodes elongation factor-G, the target of fusidic acid. Eur J Gastroenterol Hepatol, 2000 Dec, 12(12), 1323 - 8 Administering granulocyte colony-stimulating factor to acute liver failure patients corrects neutrophil defects; Rolando N et al.; OBJECTIVES: Neutrophil function is defective in acute liver failure (ALF) and the in vitro ability of granulocyte colony-stimulating factor (G-CSF) to reverse these defects has been reported . The effects of administering G-CSF to ALF patients are presented in this study . DESIGN: This was a prospective, phase I/II, open label, study . SETTING: The liver intensive therapy unit at King's College Hospital, London . PARTICIPANTS: Sequential patients admitted with acute liver failure due to acetaminophen overdose . INTERVENTIONS: G-CSF was given to four groups (each n = 6) of ALF patients as a daily infusion at 25, 50, 100 or 150 microg/m2 . A control group of eight patients did not receive G-CSF . MAIN OUTCOME MEASURES: Neutrophil phagocytosis and killing of Staphylococcus aureus and superoxide release before G-CSF administration and at 24 and 96 h thereafter . RESULTS: Neutrophils from patients receiving 50, 100 or 150 microg/m2 G-CSF, but not from control patients or those receiving 25 microg/m2, showed significantly increased phagocytosis and killing at 96 h . Doses of 50 or 150 microg/m2 G-CSF resulted in increased superoxide production at 96 h . No patients discontinued treatment as a consequence of side effects related to G-CSF administration . CONCLUSIONS: G-CSF administration is a safe and effective means of reversing the neutrophil defects of ALF, and may have a role in the prevention and treatment of infection in these patients . A dose of 50 microg/m2/day is as effective as higher doses and was associated with fewer side effects. Ned Tijdschr Geneeskd, 2000 Dec 30, 144(53), 2545 - 9 {What is to be done with vancomycin-resistant enterococcal infections?}; Bonten MJ et al.; Recently, three epidemics in Dutch hospitals were caused by vancomycin-resistant enterococci (VRE) . Although the number of infections was small, spread of colonization was extensive and many infection control measures were necessary to prevent further spread . VRE are relatively avirulent bacteria . However, few, if any, antibiotics are available for treatment of infections caused by VRE and the genetic code for resistance may be transferable to other, more virulent, bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA) . Although colonization and infection with MRSA have become endemic in many surrounding countries, such a situation has been prevented in the Netherlands by employing an aggressive 'search and destroy' policy . Although many questions regarding the optimal approach of VRE remain unanswered, a similar policy as employed for MRSA will not be possible . In contrast to MRSA, colonization with VRE occurs in the open population, no populations with increased risk for colonization appear to be definable and colonization cannot be eradicated . Based on common sense, a differentiated approach seems indicated in which extensive infection control measures should only be implemented when spread of a single genotype has been demonstrated . A reference laboratory should be created for uniform genotyping. Wound Repair Regen, 2000 Nov-Dec, 8(6), 562 - 6 Use of a noncontact radiant heat bandage and Staphylococcus aureus dermal infections in an ovine model; Lee ES et al.; Diabetic foot wounds are difficult to manage due to relative tissue ischemia and high rates of soft tissue infection . One potential treatment modality is the application of local radiant heat to promote wound healing and control infection . However, there are concerns that local heat will spread rather than control infection . We determined in this study the effect of a noncontact radiant heat bandage in controlling an ischemic soft tissue infection . Bilateral 10 x 15 cm dermal flaps were created in 15 adult range sheep . The flaps were inoculated intradermally with 107 Staphylococcus aureus in 3 separate areas . The control flap was left open to air, while the treatment flap was covered with a noncontact radiant heat bandage and heated to 38 degrees C for three 1-hour periods separated by two 1-hour nonheating periods daily . After 10 days, both dermal flaps were harvested and sent for quantitative bacteriology . Due to operative complications, 12 of 15 sheep completed the study . The heated flap temperature was significantly higher 39.2 +/- 0.5 degrees C (+/- SE) vs . the control flap 36.1 +/- 0.1 degrees C (p < 0.00001) and bacterial counts were significantly smaller in the heated flap (median 1.0 x 107 colony-forming units per gm tissue) when compared to the control flap (median 7.5 x 107) (p = 0.001) . This study shows the use of a noncontact radiant heat bandage controls ischemic soft tissue infections in an ovine model. Epilepsia, 2001 Jan, 42(1), 133 - 5 Deep wound infection after vagus nerve stimulator implantation: treatment without removal of the device; Ortler M et al.; Effective treatment of deep wound infection without removal of a previously implanted foreign body is difficult . The Neurocybernetic Prosthesis (NCP) System (Cyberonics Inc., Webster, TX, U.S.A.), implanted for vagus nerve stimulation in patients with medically refractory epilepsy, uses coil-like electrodes placed around the left vagus nerve after exposure of the nerve in the carotid sheath . Infection within this compartment endangers the contained structures and makes removal of the system hazardous . We report the case of one patient implanted with the NCP who underwent successful open wound treatment without removal of the system . A 35-year-old man had local signs of wound infection 5 weeks after implantation of a vagus nerve stimulator . Systemic signs of infection were absent . C-reactive protein was slightly elevated, but all other laboratory values were normal . After open wound debridement and thorough rinsing with bacitracin-containing solution, the wound was packed with 3% iodoformized gauze . The NCP was left in place . Systemic antibiotic therapy with fosfomycin and cefmenoxim was started . Cultures confirmed an infection with Staphylococcus aureus . The wound was rinsed daily with 3% hydrogen peroxide solution and 5% saline until cultures were sterile and granulation tissue started to fill the wound . Delayed primary closure was performed 2 weeks later . Wound healing was accomplished without removal of the device . No signs of recurrent infection were observed during a follow-up of 1 year . Open wound treatment without removal of the implanted vagus nerve stimulator is feasible in cases of deep cervical wound infection and can be an alternative if removal of the device appears hazardous. Cell Microbiol, 1999 Sep, 1(2), 101 - 17 Fibronectin-binding protein acts as Staphylococcus aureus invasin via fibronectin bridging to integrin alpha5beta1; Sinha B et al.; The ability of Staphylococcus aureus to invade mammalian cells may explain its capacity to colonize mucosa and to persist in tissues after bacteraemia . To date, the underlying molecular mechanisms of cellular invasion by S . aureus are unknown, despite its high prevalence and difficulties in treatment . Here, we show cellular invasion as a novel function for an S . aureus adhesin, previously implicated solely in attachment . S . aureus, but not S . epidermidis, invaded epithelial 293 cells in a temperature- and F-actin-dependent manner . Formaldehyde-fixed and live bacteria were equally invasive, suggesting that no active bacterial process was involved . All clinical S . aureus isolates analysed, but only a subset of laboratory strains, were invasive . Fibronectin-binding proteins (FnBPs) acted as S . aureus invasins, because: (i) FnBP deletion mutants of invasive laboratory strains lost invasiveness; (ii) expression of FnBPs in noninvasive strains conferred invasiveness; and (iii) the soluble isolated fibronectin-binding domain of FnBP (D1-D4) completely blocked invasion . Integrin alpha5beta1 served as host cell receptor, which interacted with staphylococcal FnBPs through cellular or soluble fibronectin . FnBP-deficient mutants lost invasiveness for epithelial cells, endothelial cells and fibroblasts . Thus, fibronectin-dependent bridging between S . aureus FnBPs and host cell integrin alpha5beta1 is a conserved mechanism for S . aureus invasion of human cells . This may prove useful in developing new therapeutic and vaccine strategies for S . aureus infections. Am J Epidemiol, 2001 Feb 15, 153(4), 345 - 52 Predictive 5-year survivorship model of cystic fibrosis; Liou TG et al.; The objective of this study was to create a 5-year survivorship model to identify key clinical features of cystic fibrosis . Such a model could help researchers and clinicians to evaluate therapies, improve the design of prospective studies, monitor practice patterns, counsel individual patients, and determine the best candidates for lung transplantation . The authors used information from the Cystic Fibrosis Foundation Patient Registry (CFFPR), which has collected longitudinal data on approximately 90% of cystic fibrosis patients diagnosed in the United States since 1986 . They developed multivariate logistic regression models by using data on 5,820 patients randomly selected from 11,630 in the CFFPR in 1993 . Models were tested for goodness of fit and were validated for the remaining 5,810 patients for 1993 . The validated 5-year survivorship model included age, forced expiratory volume in 1 second as a percentage of predicted normal, gender, weight-for-age z score, pancreatic sufficiency, diabetes mellitus, Staphylococcus aureus infection, Burkerholderia cepacia infection, and annual number of acute pulmonary exacerbations . The model provides insights into the complex nature of cystic fibrosis and supplies a rigorous tool for clinical practice and research. Vet Microbiol, 2001 Feb 26, 78(4), 373 - 82 Characterization of enterotoxigenic Staphylococcus aureus strains isolated from bovine mastitis in north-east Switzerland; Stephan R et al.; Thirty-four strains of enterotoxin-producing Staphylococcus aureus obtained from milk samples of 34 dairy cows suffering from mastitis from 34 different farms in north-east Switzerland were identified and further characterized by pheno- and genotypic methods . This included the identification of staphylococcal enterotoxin (SE) types, an antibiotic resistance testing, the appraisal of hemolysis, the egg yolk reaction, the detection of the clumping factor and protein A by means of a latex agglutination, the PCR amplification of a S . aureus specific part of the gene encoding the 16S-23S rRNA "intergenic spacer" region and a species specific part of the 23S rRNA-gene, the PCR amplification of the clumping factor (clfA) gene, the X region and the IgG-binding region of the protein A (spa) gene, the coagulase (coa) gene and additionally a macrorestriction analysis of the chromosomal DNA . Within the 26 cultures which formed a single SE, there were 23 SEC- and three SED-formers . Eight cultures were SEAD formers . It was remarkable that 22 SEC formers were also positive for TSST-1 . Eighteen of the 23 SEC-formers could be classified as being of the same phenotype . Most of the cultures of one enterotoxin type also showed a great uniformity in the size and number of repeats of the X region as well as in the size of the IgG-binding region of protein A gene and in the size of the coagulase gene . Macrorestriction analysis revealed 11 PFGE patterns . These were in part only different from each other in a few fragments and thus displayed close clonal relations . The results of the present investigation show that a broad distribution of identical or closely related enterotoxin-producing S . aureus clones seem to contribute to the bovine mastitis problem in north-east Switzerland. Urology, 2001 Feb, 57(2), 246 - 51 Diagnosis and treatment of pyogenic psoas abscess in diabetic patients: usefulness of computed tomography and gallium-67 scanning; Kao PF et al.; OBJECTIVES: To examine retrospectively the clinical presentations, microbiologic characteristics, and treatment outcomes of psoas abscess in patients with diabetes mellitus (DM) and to assess the usefulness of computed tomography and gallium-67 scanning in its early diagnosis . METHODS: During a 9-year period, psoas abscesses in patients with DM were collected at a medical center . The clinical history and associated etiologic factors, microbiologic results, clinical outcomes, and hospitalization days were recorded . The use of imaging in the diagnosis of psoas abscess and other concomitant infectious lesions was also studied . RESULTS: Fifteen patients with DM and psoas abscess (13 women and 2 men; mean age 58.7 +/- 9.0 years) were found . The most frequent symptom was fever (12 of 15) . Of the six different microorganisms that grew in the blood and/or abscess cultures, Staphylococcus aureus was the most frequent (7 of 15) . The most commonly associated pathologic finding was vertebral osteomyelitis (5 of 15) . Computed tomography and/or magnetic resonance imaging confirmed the diagnosis of psoas abscesses in all 15 patients . The gallium-67 scan especially aided in the diagnosis of the patients who had initially been diagnosed as having fever of unknown origin (4 of 5) and in the diagnosis of concomitant lesions (9 of 12) . Debridement or surgical drainage of the abscess was done in 12 patients . All the patients received adequate antibiotic treatment . However, the mortality rate was 20% . The average hospitalization stay was 42.7 +/- 20.7 days . CONCLUSIONS: Psoas abscess in patients with DM is a disease with both diagnostic and therapeutic challenges . We found the infecting microorganisms to be variable and the mortality rate high. Antimicrob Agents Chemother, 2001 Mar, 45(3), 927 - 31 Relationships of the area under the curve/MIC ratio to different integral endpoints of the antimicrobial effect: gemifloxacin pharmacodynamics in an in vitro dynamic model; Firsov AA et al.; Most integral endpoints of the antimicrobial effect are determined over an arbitrarily chosen time period, such as the dosing interval (tau), regardless of the actual effect duration . Unlike the tau-related endpoints, the intensity of the antimicrobial effect (I(E)) does consider its duration-from time zero to the time when bacterial counts on the regrowth curve achieve the same maximal numbers as in the absence of the antimicrobial . To examine the possible impact of this fundamental difference on the relationships of the antimicrobial effect to the ratio of the area under the concentration-time curve (AUC) to the MIC, a clinical isolate of Staphylococcus aureus was exposed to simulated gemifloxacin pharmacokinetics over a 40-fold range of AUC/MIC ratios, from 11 to 466 h . In each run, I(E) and four tau-related endpoints, including the area under the time-kill curve (AUBC), the area above the curve (AAC), the area between the control growth and time-kill curves (ABBC), and the ABBC related to the area under the control growth curve (AUGC), were calculated for tau = 24 h . Unlike the I(E), which displayed pseudolinear relationships with the AUC/MIC ratio; each tau-related endpoint showed a distinct saturation at potentially therapeutic AUC/MIC ratios (116 to 466 h) when the antimicrobial effect persisted longer than tau . This saturation results from the underestimation of the true effect and may be eliminated if ABBC, AAC, and AUBC (but not AUGC) are modified and determined in the same manner as the I(E) to consider the actual effect duration . These data suggest a marginal value of the tau-related endpoints as indices of the total antimicrobial effect . Since all of them respond to AUC/MIC ratio changes less than the I(E), the latter is preferable in comparative pharmacodynamic studies. Antimicrob Agents Chemother, 2001 Mar, 45(3), 815 - 24 Eagle-type methicillin resistance: new phenotype of high methicillin resistance under mec regulator gene control; Kondo N et al.; We report a novel phenotype of methicillin resistance, designated "Eagle-type" resistance, which is characteristic in its resistance to high concentrations of methicillin (64 to 512 microg/ml) and susceptibility to low concentrations of methicillin (2 to 16 microg/ml) . The type of resistance was expressed in mutant strains selected with high concentrations (e.g., 128 to 512 microg/ml) of methicillin from the pre-methicillin-resistant Staphylococcus aureus strain N315, whose mecA gene transcription is strongly repressed by the mecI gene-encoded repressor protein MecI . The Eagle-type mutant strains harbored no mutation in the mecI gene or in the operator region of mecA gene to which MecI repressor is supposed to bind . In the representative Eagle-type strain h4, repression of mecA gene transcription and penicillin-binding protein 2' production were found to be released by exposing the cells to a high concentration (128 microg/ml) of methicillin but not to lower concentrations (1 and 8 microg/ml) of methicillin . The strain h4 expressed paradoxical susceptibility (Eagle effect) to the cytokilling activity of methicillin . Experimental deletion of mecI gene from the chromosome of h4 by mecI-specific gene substitution converted its Eagle-type resistance to homogeneously high methicillin resistance . We cloned two novel genes, designated hmrA and hmrB, from genomic library of h4, which conferred Eagle-type resistance to N315 when introduced into the cell in multiple copies . The genes were shown to confer homogeneous methicillin resistance to the heterogeneously methicillin-resistant strain LR5 when they were introduced into on multicopy plasmids . This result strongly indicated that the genetic alteration responsible for the expression of the Eagle phenotype is identical, or equivalent in its effect, to the genetic alteration underlying heterogeneous-to-homogeneous conversion of methicillin resistance in S . aureus. Clin Infect Dis, 2001 Feb 15, 32(4), 647 - 9 Epub 2001 Feb 07. Infection of orthopedic prostheses after Staphylococcus aureus bacteremia; Murdoch DR et al.; We prospectively evaluated 53 patients with prosthetic joints and 27 patients with other orthopedic prosthetic devices who developed Staphylococcus aureus bacteremia (SAB) . After exclusion of patients with primary postoperative infections, the risk of a prosthesis becoming infected by means of hematogenous seeding after SAB was 34% (15 of 44 patients) for prosthetic joints and 7% (1 of 15 patients) for other orthopedic prostheses. Clin Infect Dis, 2001 Feb 15, 32(4), 581 - 6 Epub 2001 Feb 06. Epidemic spread of a single clone of methicillin-resistant Staphylococcus aureus among injection drug users in Zurich, Switzerland; Fleisch F et al.; We describe an outbreak of methicillin-resistant Staphylococcus aureus (MRSA) among injection drug users (IDUs) . From August 1994 through December 1999, we registered 31 IDUs with MRSA infections (12 with soft-tissue infection, 7 with pneumonia {fatal in 1}, 7 with endocarditis {fatal in 1}, 2 with osteomyelitis, 2 with septic arthritis, and 1 with ulcerative tonsillitis), with a marked increase in the number of IDUs registered during 1998 and 1999 . Of 31 patients, 15 (48%) were infected with human immunodeficiency virus . A point-prevalence study among IDUs who frequented outpatient facilities in Zurich revealed an MRSA carriage rate of 10.3% (range, 0%-28.6%) in various facilities . In all but 1 case, pulsed-field gel electrophoresis banding patterns of isolates obtained from these patients were indistinguishable from isolates of the initial 31 IDUs registered . Risk factors for MRSA carriage were disability and prior hospitalization in a hospice . In summary, MRSA became endemic in IDUs in Zurich as a result of the spread of a single clone . This clone caused major morbidity and was responsible for a lethal outcome in 2 cases. J Surg Res, 2001 Mar, 96(1), 35 - 43 A mouse model for postoperative fatal enteritis due to Staphylococcus infection; Nakamura Y et al.; BACKGROUND: Postoperative infection of intestine with methicillin-resistant Staphylococcus aureus (MRSA) is fatal in some cases . The object of this study was to establish a mouse model for the infection, providing a useful tool for investigating mechanisms in the progression of infection . METHODS: Mice were pretreated with cyclophosphamide, injected orally or directly into jejunum with MRSA prepared from a postoperative patient, and then given 5 daily doses of antibiotics . Forty-eight hours after the injection, bacterial translocation and serum endotoxin levels were examined . Macrophage depletion was carried out by the administration of liposome-encapsulated dichloromethylene diphosphate (Cl(2)MDP), 4 days before MRSA injection . RESULTS: Injection into the jejunum but not oral administration of MRSA induced enteritis with diarrhea and resulted in death in most cyclophosphamide-treated mice . Translocation of MRSA in mesenteric lymph nodes and liver was observed, concomitantly with E . coli infection . Endotoxin-resistant C3H/HeJ mice infected with MRSA survived longer than endotoxin-sensitive C3H/He mice, but also died within a week after MRSA injection . Selective depletion of macrophages induced infection in mice that were not pretreated with cyclophosphamide . CONCLUSION: We established a mouse model for the fatal MRSA infection which induced enteritis with diarrhea, that will be a useful tool for investigating the mechanisms for sometimes fatal MRSA infection of the intestine in postoperative patients . The presence of E . coli or endotoxin seemed to play a major role in the mortality of mice in the early days of MRSA-induced enteritis, but other factors, probably from MRSA, in the later days . Phagocytes were quite important for protection against the MRSA infection . Infect Immun, 2001 Mar, 69(3), 1957 - 60 Pheromone cross-inhibition between Staphylococcus aureus and Staphylococcus epidermidis; Otto M et al.; Cross-inhibition by quorum-sensing pheromones between Staphylococcus aureus and Staphylococcus epidermidis was investigated using all known S . aureus agr pheromone subgroups . All S . aureus subgroups were sensitive towards the S . epidermidis pheromone, with the exception of the recently identified subgroup 4 . The subgroup 4 pheromone was also the only S . aureus pheromone able to inhibit the S . epidermidis agr response . The close relation of subgroup 4 to subgroup 1 suggests that subgroup 4 might have evolved from subgroup 1 by mutation under the selective pressure of competition with S . epidermidis . The competition between S . aureus and S . epidermidis by means of quorum-sensing cross talk seems to be generally in favor of S . epidermidis, which might explain the predominance of S . epidermidis on the skin and in infections on indwelling medical devices. Infect Immun, 2001 Mar, 69(3), 1521 - 7 Molecular characterization of a novel Staphylococcus aureus serine protease operon; Reed SB et al.; The present study identified and characterized a unique operon (spl) encoding six serine protease-like proteins . In addition, native Spl proteins were isolated and characterized . Typical of most exoproteins, the spl gene products contain putative 35- or 36-amino-acid signal peptides . The Spl proteins share 44 to 95% amino acid sequence identity with each other and 33 to 36% sequence identity with V8 protease . They also contain amino acids found in catalytic triads of enzymes in the trypsin-like serine protease family, and SplB and SplC were shown to degrade casein . The spl operon is transcribed on a 5.5-kb transcript, but several nonrandom degradation products of this transcript were also identified . Similar to other S . aureus exoprotein genes, the spl operon is maximally expressed during the transition into stationary phase and is positively controlled by the Agr virulence factor regulator . The Sar regulatory system did not affect spl operon expression . PCR analysis revealed the presence of the spl operon in 64% of the S . aureus isolates tested, although one spl operon-negative isolate was shown to contain at least two of the spl genes . Finally, intraperitoneal injection of an spl operon deletion mutant revealed no major differences in virulence compared to the parental strain. Infect Immun, 2001 Mar, 69(3), 1469 - 76 Human lactoferrin and peptides derived from its N terminus are highly effective against infections with antibiotic-resistant bacteria; Nibbering PH et al.; Since human lactoferrin (hLF) binds to bacterial products through its highly positively charged N terminus, we investigated which of the two cationic domains is involved in its bactericidal activity . The results revealed that hLF lacking the first three residues (hLF(-3N)) was less efficient than hLF in killing of antibiotic-resistant Staphylococcus aureus, Listeria monocytogenes, and Klebsiella pneumoniae . Both hLF preparations failed to kill Escherichia coli O54 . In addition, hLF(-3N) was less effective than hLF in reducing the number of viable bacteria in mice infected with antibiotic-resistant S . aureus and K . pneumoniae . Studies with synthetic peptides corresponding to the first 11 N-terminal amino acids, designated hLF(1-11), and fragments thereof demonstrated that peptides lacking the first three N-terminal residues are less effective than hLF(1-11) in killing of bacteria . Furthermore, a peptide corresponding to residues 21 to 31, which comprises the second cationic domain, was less effective than hLF(1-11) in killing of bacteria in vitro and in mice having an infection with antibiotic-resistant S . aureus or K . pneumoniae . Using fluorescent probes, we found that bactericidal hLF peptides, but not nonbactericidal peptides, caused an increase of the membrane permeability . In addition, hLF killed the various bacteria, most probably by inducing intracellular changes in these bacteria without affecting the membrane permeability . Together, hLF and peptides derived from its N terminus are highly effective against infections with antibiotic-resistant S . aureus and K . pneumoniae, and the first two arginines play an essential role in this activity. Infect Immun, 2001 Mar, 69(3), 1287 - 97 Fas (CD95)-Fas ligand interactions are responsible for monocyte apoptosis occurring as a result of phagocytosis and killing of Staphylococcus aureus; Baran J et al.; Human peripheral blood monocytes become apoptotic following phagocytosis of Staphylococcus aureus . In this study, we investigated the mechanisms involved in this phenomenon . Cells exposed to bacteria were examined for the surface expression of Fas and Fas ligand (FasL) . The level of soluble form of FasL was also measured in the culture supernatants . As Fas-mediated apoptosis involves the activation of caspases, the activities of caspase-8 and caspase-3 were determined . Finally, the involvement of oxidative stress in apoptosis of infected monocytes was investigated . The data indicated that as a consequence of phagocytosis of S . aureus, FasL is released from the monocyte surface and induces apoptosis of phagocytic monocytes and to some extent the bystander cells . The importance of this mechanism was confirmed by demonstrating that blockage of CD95 prevents S . aureus-induced apoptosis of monocytes . Cell death occurring after phagocytosis of S . aureus involves the activation of caspase-3-like proteases, as the specific caspase-3 inhibitor suppressed apoptosis of infected cells . The generation of reactive oxygen intermediates by phagocytic monocytes by itself is not sufficient as a death signal but rather acts in up-regulating FasL shedding and possibly in modulating caspase activity. Heart . 2001 Mar;85(3):E4. Cardiac rupture caused by Staphylococcus aureus septicaemia and pericarditis: an incidental finding; Osula S et al.; A 35 year old woman with a long history of intravenous drug abuse presented to a local hospital with severe anaemia, fever, raised markers of inflammation, and positive blood cultures for Staphylococcus aureus . She responded to treatment with antibiotics with improvement in her symptoms and markers of inflammation . Four weeks later a "routine" echocardiogram showed a rupture of her left ventricular apex and a large pseudoaneurysm . There had been no deterioration in her symptoms or haemodynamic status to herald this new development . It was successfully repaired surgically and the patient made a good recovery. Arthritis Res, 2001, 3(1), 41 - 7 Epub 2000 Nov 03. Role of IL-12 in Staphylococcus aureus-triggered arthritis and sepsis; Hultgren OH et al.; The present study demonstrates that endogenous production of IL-12 is crucial for survival in Staphylococcus aureus-induced arthritis in mice . Staphylococcal load is enhanced in several organs, because of lack of IL-12 . This might be due to decreased production of IFN-gamma in IL-12-deficient mice . Although IL-12-deficient mice were exposed to higher staphylococcal load, they demonstrated no increased severity of arthritis as compared with control animals. Crit Care Med, 2001 Jan, 29(1), 1 - 7 Biosynthesis of constitutive nitric oxide synthase-derived nitric oxide attenuates coronary vasoconstriction and myocardial depression in a model of septic heart failure induced by Staphylococcus aureus alpha-toxin; Grandel U et al.; OBJECTIVE: Myocardial depression, which frequently occurs in the course of septic shock, has been attributed to the cardiodepressant properties of nitric oxide (NO) generated by either the inducible NO synthase (iNOS) or the constitutive isoform (cNOS) . We have previously demonstrated that alpha-toxin from Staphylococcus aureus induces thromboxane-mediated vasoconstriction accompanied by severe cardiodepression in isolated rat hearts . In the present study, we investigated the role of NO in the alpha-toxin-induced vascular and contractile abnormalities . DESIGN: Prospective, experimental study . SETTING: Research laboratory at a university hospital . SUBJECTS: Isolated hearts from male Wistar rats . INTERVENTIONS: Isolated hearts were perfused with purified staphylococcal alpha-toxin for 60 mins . MEASUREMENTS AND MAIN RESULTS: At a concentration of 0.25 and 0.5 microg/mL, alpha-toxin induced a rise in coronary perfusion pressure, depressed myocardial contractility, and caused edema formation . Simultaneously, a time- and dose-dependent rapid release of NO into the perfusate was noted as quantified by a chemiluminescence technique . L-NMMA, a nonselective inhibitor of NOS, but not PBITU, an iNOS-selective inhibitor, blocked NO synthesis, markedly increased the rise in coronary perfusion pressure and the loss in contractility, and enhanced edema formation in response to alpha-toxin . In contrast, zaprinast, a selective inhibitor of phosphodiesterase type V that is used for stabilization of cyclic guanosine monophosphate, attenuated the toxin-induced coronary vasoconstrictor response and the myocardial depression . L-arginine, the substrate of NOS, had similar, yet less potent, effects as zaprinast and slightly increased the release of NO caused by alpha-toxin . Immunohistochemical analysis of the myocardium at the end of the perfusion period demonstrated a positive staining for cNOS but not for iNOS . In addition, no up-regulation of iNOS mRNA was detected in the tissue of toxin-exposed hearts . CONCLUSIONS: Staphylococcal alpha-toxin provokes NO biosynthesis via activation of cNOS in rat hearts . NO partly antagonizes the deleterious effects of this pathogenicity factor on coronary vasoregulation and myocardial performance. Rev Saude Publica, 2000 Dec, 34(6), 578 - 80 {Occurrence of Staphylococcus aureus in "frescal" type cheese}; de Almeida Filho E et al.; OBJECTIVE: To verify the occurrence of Staphylococcus aureus in a sample of cheese sold in a city of the Southeastern region of Brazil and assess the potential risk for the consumers . METHODS: Eighty samples of homemade cheese sold in a city of the southern region of Brazil were evaluated for the presence and the most probable number of Staphylococcus aureus agents . RESULTS: The study revealed the presence of S . aureus in 40 samples (50.0%, with a mean count of 10(5)/g . CONCLUSION: These results are worrisome because the Health Ministry has established a safety threshold of 10(3)/g, and the values obtained are close to the number of bacteria able to produce enough enterotoxins to cause a foodborne disease outbreak. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2001 Jan, 91(1), 85 - 8 Common variable immunodeficiency with unusual vegetative lesions of the tongue and labial mucosa: a case report; Kishimoto H et al.; A case of common variable immunodeficiency with unusual vegetative lesions of the tongue and lower lip in a 28-year-old man is presented . The vegetative lesions developed over the preceding 10 months and clinically were suggestive of malignancy . The biopsy specimens showed no malignancy, and a bacterial culture of the tongue detected abundant Staphylococcus aureus . Combined treatment with a corticosteroid antibiotic ointment and povidone iodine rinse produced remarkable resolution of the lesions . Laboratory examination showed markedly decreased levels of serum immunoglobulins . Intravenous gamma globulin replacement therapy resulted in good control of infection and disappearance of the lesions. Ann Thorac Cardiovasc Surg, 2000 Dec, 6(6), 414 - 7 Successful treatment of MRSA mediastinitis after aortic arch replacement; Kaneda T et al.; An aortic arch graft replacement was successfully performed for a true aneurysm on the distal aortic arch . However, Methicillin-resistant Staphylococcus aureus (MRSA) mediastinitis occurred after surgery . Following reoperation for debridement and irrigation of the mediastinal space, antibiotics were administered and continuous irrigation of mediastinal space with saline containing appropriate antibiotics and intermittently short duration irrigation with a large quantity of saline containing popidone-iodine were carried out with good results . This paper presents the successful treatment of mediastinitis after replacement of the aortic arch without removing the graft, followed by a brief review of the literature regarding infection of prostetic grafts for the thoracic aorta. Diagn Microbiol Infect Dis, 2001 Jan, 39(1), 39 - 47 Comparative in vitro pharmacodynamics of imipenem and meropenem against ATCC strains of Escherichia coli, Staphylococcus aureus and Bacteroides fragilis; White RL et al.; We evaluated the pharmacodynamics of imipenem and meropenem, utilizing time-kill studies over a concentration/MIC (C/MIC) range of 0.0625-1024 for E . coli ATCC 35218 (EC) and S . aureus ATCC 29213 (SA) and from 0.125-512 for B . fragilis ATCC 25285 (BF) . Area under the time-kill curves were converted to percent response (%R) . AUCs were calculated from drug concentrations corrected for degradation and %R vs . C/MIC and AUC/MIC were fit to a sigmoidal Emax model . Emax was similar for both agents for all organisms . Meropenem was 4x more potent than imipenem against EC based on the MIC and required 7 and 13.5-fold lower AUCs to achieve E50 and E90, (50% and 90% of the maximal response, respectively) respectively, whereas imipenem was 8x more potent than meropenem against SA based on the MIC and required 8 and 13-fold lower AUCs to achieve E50 and E90, respectively . The C/MIC and AUC/MIC to achieve E50 and E90 with imipenem were 2- and fourfold higher, respectively than meropenem against EC . There was less than a twofold difference in C/MICs and AUC/MIC between imipenem and meropenem for both E50 and E90 against SA . Against BF, concentrations and AUCs at E50 were similar for both agents; however, imipenem required 4 to 6-fold higher concentrations and AUC to achieve E90 . Although there are differences in the potency of these agents as assessed by the MIC or AUC vs . response, when normalized by the MIC and corrected for drug degradation, these agents displayed similar pharmacodynamic parameter-response relationships. Am J Infect Control, 2001 Feb, 29(1), 13 - 9 Community-acquired bacteremia at a teaching versus a nonteaching hospital: impact of acute severity of illness on 30-day mortality; Mylotte JM et al.; BACKGROUND: Few studies have focused recently on the epidemiology of community-acquired bacteremia (CAB) and there have been few comparisons of CAB in teaching versus nonteaching hospitals . OBJECTIVES: To compare the clinical characteristics, acute severity of illness, and 30-day mortality of patients with CAB admitted to a teaching and a nonteaching hospital and to define predictors of 30-day mortality among patients with CAB that would be identifiable at the time of admission to the hospital . METHODS: This was a retrospective study of CAB at a teaching hospital (n = 174 episodes) compared to a community nonteaching hospital (n = 74 episodes) during 1995 . Data collected included demographic characteristics, underlying diseases, sources of CAB, and antimicrobial therapy . Acute severity of illness on admission was measured by using the acute physiology score component of the Acute Physiology and Chronic Health Evaluation III system (APS APACHE III).Main Outcome Measure: Status, dead or alive, 30 days after admission for CAB . RESULTS: At the nonteaching hospital, patients were older but, on average, significantly less acutely ill (as measured by the admission APS APACHE III score) than were those at the teaching hospital . In contrast, patients with HIV infection, posttransplantation, or on hemodialysis were identified only at the teaching hospital . Overall, organisms causing CAB at both hospitals were similar except that Staphylococcus aureus CAB occurred significantly more often at the teaching hospital and Escherichia coli CAB occurred more often at the nonteaching hospital . There was no significant difference in 30-day mortality in patients with CAB between the teaching hospital (19.3%) and the nonteaching hospital (16.7%; P =.63) . APS APACHE III score on admission identified episodes of CAB with a low- and a high-risk for 30-day mortality at both hospitals . Independent predictors of 30-day mortality were APACHE III score on admission (P <.001) and pneumonia as a source of CAB (P =.012) . CONCLUSIONS: Among patients with CAB, acute severity of illness on admission was the most important predictor of 30-day mortality at both hospitals . Even though patients with CAB were, on average, more severely ill at the time of admission to the teaching hospital, 30-day mortality rates were not significantly different between the two hospitals because deaths correlated with high APS APACHE III scores at both facilities . The APS APACHE III score on admission provides important prognostic information among patients with CAB. Clin Infect Dis, 2001 Feb 1, 32(3), E53 - 6 Epub 2001 Jan 24. Bacteremic nonmenstrual staphylococcal toxic shock syndrome associated with enterotoxins A and C; Czachor J et al.; We report a case of bacteremic nonmenstrual staphylococcal toxic shock syndrome (STSS) producing staphylococcal enterotoxins A and C . The bloodstream isolation of Staphylococcus aureus, as well as the production of enterotoxins A and C, are unusual as separate entities, and distinctly uncommon when found together. Clin Infect Dis, 2001 Feb 1, 32(3), 402 - 12 Epub 2001 Jan 26. Linezolid (PNU-100766) versus vancomycin in the treatment of hospitalized patients with nosocomial pneumonia: a randomized, double-blind, multicenter study; Rubinstein E et al.; Linezolid, the first oxazolidinone, is active against gram-positive bacteria, including multidrug-resistant strains . This multinational, randomized, double-blind, controlled trial compared the efficacy, safety, and tolerability of linezolid with vancomycin in the treatment of nosocomial pneumonia . A total of 203 patients received intravenous linezolid, 600 mg twice daily, plus aztreonam, and 193 patients received vancomycin, 1 g intravenously twice daily, plus aztreonam for 7-21 days . Clinical and microbiological outcomes were evaluated at test of cure 12-28 days after treatment . Clinical cure rates (71 {66.4%} of 107 for linezolid vs . 62 {68.1%} of 91 for vancomycin) and microbiological success rates (36 {67.9%} of 53 vs . 28 {71.8%} of 39, respectively) for evaluable patients were equivalent between treatment groups . Eradication rates of methicillin-resistant Staphylococcus aureus and safety evaluations were similar between treatment groups . Resistance to either treatment was not detected . Linezolid is a well-tolerated, effective treatment for adults with gram-positive nosocomial pneumonia. Clin Infect Dis, 2001 Feb 1, 32(3), 367 - 72 Epub 2001 Jan 18. Effect of hand cleansing with antimicrobial soap or alcohol-based gel on microbial colonization of artificial fingernails worn by health care workers; McNeil SA et al.; This study was undertaken to determine differences in microflora on the nails of health care workers (HCWs) wearing artificial nails compared with control HCWs with native nails and to assess the effect on these microflora of hand cleansing with antimicrobial soap or alcohol-based gel . Cultures were obtained from 21 HCWs wearing artificial nails and 20 control HCWs before and after using antimicrobial soap or alcohol-based gel . Before cleansing with soap, 86% of HCWs with artificial nails had a pathogen (gram-negative bacilli, Staphylococcus aureus, or yeasts) isolated, compared with 35% of controls (P=.003); a similar difference was noted before hand cleansing with gel (68% vs . 28%; P=.03) . Significantly more HCWs with artificial nails than controls had pathogens remaining after hand cleansing with soap or gel . Of HCWs with artificial nails, only 11% cleared pathogens with soap compared with 38% with gel . Of control HCWs, only 14% cleared with soap compared with 80% with gel . Artificial acrylic fingernails could contribute to the transmission of pathogens, and their use by HCWs should be discouraged. J Hosp Infect, 2001 Feb, 47(2), 104 - 9 A hospital-acquired outbreak of methicillin-resistant Staphylococcus aureus infection initiated by a surgeon carrier; Wang JT et al.; Methicillin-resistant Staphylococcus aureus (MRSA) has become an important hospital-acquired pathogen, infection with which often leads to major morbidity and mortality . The principal mode of transmission for MRSA is transfer of the organism from a carrier or infected patient to uninfected patients by the hands or clothing of staff . From January 16 1997 to April 2 1997, five patients who had undergone open-heart surgery in a hospital located in northern Taiwan, developed surgical wound infections and mediastinitis caused by MRSA . All patients were hospitalized in two adjacent surgical intensive care units (ICUs) following their respective operations . Consequently, the hospital's infection control team commenced investigation of the outbreak . Pulsed-field gel electrophoresis (PFGE) has been shown to be a good technique for epidemiological typing . By analysing cultures taken from staff by PFGE, it was demonstrated that this outbreak was most likely to be initiated by a surgeon with MRSA carriage . After elimination of the carrier state using topical mupirocin treatment, the outbreak was controlled without further incident . J Hosp Infect, 2001 Feb, 47(2), 98 - 103 MRSA colonization and the risk of MRSA bacteraemia in hospitalized patients with chronic ulcers; Roghmann MC et al.; A cohort study of patients with chronic ulcers was performed to estimate the risk of methicillin-resistant Staphylococcus aureus (MRSA) bacteraemia in a population colonized with MRSA . During a five-year period (January 1990-May 1995), 911 patients with chronic ulcers (CU), as determined by ICD9-CM code search, were admitted to an acute care hospital . Sixty percent (545/911) of these patients with CU had their CU cultured to detect MRSA and 30% (166/545) of these were colonized with MRSA . Among patients with surveillance cultures, those with MRSA colonization had significantly more days of hospitalization and were also more likely to have a central venous catheter during hospitalization compared with patients without MRSA colonization . MRSA bacteraemia occurred in 4% (36/911) of CU patients during the study period and in 6% (32/545) of cultured CU patients . Among the 545 patients who had surveillance cultures, the risk ratio for MRSA bacteraemia when there was MRSA colonization of their chronic ulcer was 16 (95% CI 6-45) . Among patients with MRSA colonization, central venous catheter use was the only significant risk factor for MRSA bacteraemia . In 16 of the 28 patients with MRSA bacteraemia and MRSA colonization, the MRSA colonization was identified more than seven days before the bacteraemia . This cohort study identifies MRSA colonized CU patients in an acute care setting as a high-risk population for MRSA bacteraemia . J Hosp Infect, 2000 Dec, 46(4), 280 - 7 Antibiotic susceptibility and genotypic characterization of methicillin-resistant Staphylococcus aureus strains in eastern France; Bertrand X et al.; We studied changes over four years in the characteristics of methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in a French university hospital by analysis of antibiotic susceptibility and restriction fragment length polymorphism using pulsed field gel electrophoresis . Changes were considered in relation to regional and national frameworks . The proportion of gentamicin-susceptible methicillin-resistant S . aureus (GS-MRSA) among MRSA increased from 22.6% in 1994 to 86.8% in 1998 . We first isolated strains of GS-MRSA susceptible to tobramycin and amikacin (TKS-MRSA) in 1996 from patients in eight wards . The number of TKS-MRSA strains per 100 MRSA was 6.1 in 1997, 14.5 in 1998, and 18.9 in 1999 . Pattern A, the major DNA pattern identified, accounted for 78.6% of isolates in 1998 . This pattern was found in TKS-MRSA and TKR-MRSA strains, but not in Gentamicin-resistant MRSA strains (GR-MRSA) . Fitness analysis showed that GR-MRSA strains had much lower replication rates than GS-MRSA strains, but there was no difference between TKS-MRSA and TKR-MRSA strains . Aminoglycoside consumption remained constant between 1994 and 1998 . The spread of TKS-MRSA in our hospital since 1996 may relate to the "total use threshold", as the level of tobramycin/amikacin use is below that required for selection of TKR-MRSA . J Hosp Infect, 2000 Dec, 46(4), 271 - 9 A purpose built MRSA cohort unit; Fitzpatrick F et al.; The control of hospital-acquired infection, in particular methicillin-resistant Staphylococcus aureus (MRSA) remains a challenge . Our hospital has established a purpose built 11-bed cohort unit with on-site rehabilitation for care of patients colonized with MRSA, in an attempt to improve their quality of care . Prior to the opening of this unit a number of concerns were voiced and the aim of this study was to address these . First, to establish if patient cohorting reduces the likelihood of successful decolonization, second, to evaluate the risk of staff colonization, and finally to see if successful environmental control of MRSA is possible.A patient database was established detailing patient demographics, infection rates, eradication and reacquisition rates . Staff screening was performed weekly, at the start of a period of duty . Sixty environmental sites were screened before unit opening, at 48h, six weeks and at six months.There were 88 admissions in the first six months; 62 patients were colonized with MRSA, and 26 patients (10 surgical, 16 medical) had MRSA infections . Twenty-three of 88 patients (26%) were successfully decolonized, which compares favourably with an eradication rate of 20% for the rest of the hospital . Twenty staff members participated in weekly screening . Five staff members colonized with MRSA were detected and all were successfully decolonized . Environmental control was achieved with a combination of a daily detergent clean and a once weekly clean with phenolic disinfectant.Our preliminary data suggest that, despite cohorting patients colonized with MRSA, with proper education and supervised cleaning protocols, it is possible to control environmental MRSA load, successfully decolonize patients and limit the risk of staff colonization . J Hosp Infect, 2000 Dec, 46(4), 263 - 70 Methicillin-resistant Staphylococcus aureus in a teaching hospital: investigation of nosocomial transmission using a matched case-control study; Dziekan G et al.; In early 1996 a hospital-wide methicillin-resistant Staphylococcus aureus (MRSA) epidemic was recognized in a 900-bed university hospital . In order to investigate hospital-specific transmission routes, a case-control study was carried out . Cases and controls were matched for age (+/- 10 years), sex, admission date (+/- 10 days) and clinical department on admission . Data on potential risk factors, were retrieved by chart review . Between June 1996 and February 1997, 67 patients with hospital-acquired MRSA were identified . Molecular typing showed that 85% of the cases carried an indistinguishable strain . The average time at risk for cases and controls was 17.3 and 23.7 days, respectively (P= 0.01) . Seventeen patients (25.4%) developed infection . Conditional multivariate regression analysis showed that intensity of care (P= 0.002), number of transfers (P= 0.019), and fluoroquinolone therapy (P= 0.025) were independently associated with acquisition of MRSA . Intensity of care can be considered as a surrogate marker for a number of manipulations which represent the main risk factors for MRSA transmission . Frequent transfers within the hospital hinder, not only the epidemiological analyses, but also efforts to bring an outbreak under control . Our findings give epidemiological support to recent molecular studies which suggest that fluoroquinolone use may increase the transmissibility of MRSA in hospitals . J Med Chem, 2001 Jan 18, 44(2), 261 - 8 Flavonolignan and flavone inhibitors of a Staphylococcus aureus multidrug resistance pump: structure-activity relationships; Guz NR et al.; Although some progress has been reported on structure-activity relationships (SARs) for inhibitors of mammalian P-glycoprotein MDR efflux pumps, there is almost nothing in the literature regarding SARs for inhibitors of any bacterial efflux pump . Indeed, only a few of these have been described . Our discovery of a potent naturally occurring flavonolignan inhibitor of the NorA MDR pump of Staphylococcus aureus provided a structural foundation upon which SARs could be assessed via synthetic analogues . Several flavonolignans were prepared which proved to have greater potency than the natural isolate, 5'-methoxyhydnocarpin-D, while others showed decreased potency . Surprisingly, some simple alkylated flavones also were quite active MDR pump inhibitors . Variability of activity among the compounds tested was sufficient so that at least some SARs could be postulated and compared with those known for P-glycoprotein. J Pept Res, 2001 Feb, 57(2), 127 - 39 Increased antibacterial activity of 15-residue murine lactoferricin derivatives; Strom MB et al.; LFM W8 is a synthetic 15-residue lactoferricin derivative (H2N-EKCLRWQWEMRKVGG-COOH), corresponding to residues 16-30 of the mature murine lactoferrin protein except that the asparagine residue in position 8 of the native peptide is replaced with tryptophan . We have previously reported that the two tryptophan residues in positions 6 and 8 are of crucial importance for the antibacterial activity of many lactoferricin derivatives but, despite fulfilling this requirement, LFM W8 is inactive against Escherichia coli and Staphylococcus aureus . In order to solve this puzzle, a quantitative structure-antibacterial activity relationship study of synthetic LFM W8 derivatives was performed by replacing the glutamate residues in positions 1 and 9 with arginine or alanine, and the valine residue in position 13 with tyrosine . The results from the study were analyzed using multivariate data analysis . The derived mathematical model clustered the peptides into distinct groups which reflected their antibacterial activities, pointed out correlations between different structural parameters, highlighted the structural parameters that were important for antibacterial activity, and enabled us to predict the activity of a 15-residue bovine lactoferricin derivative . The results showed that net charge and micelle affinity, as determined from the ratio of alpha-helicity in sodium dodecyl sulfate micelles and in 1,1,1,3,3,3-hexafluoro-2-propanol, were the most important structural parameters affecting antibacterial activity . The most active derivative, LFM R1,9 W8 Y13, displayed a minimal inhibitory concentration of 10 and 12 microM against E . coli and S . aureus, respectively . This represented more than 50-fold and 40-fold increases in antibacterial activity, respectively, compared with LFM W8. Int J Dermatol, 2000 Dec, 39(12), 942 - 4 Topical gentian violet for cutaneous infection and nasal carriage with MRSA; Okano M et al.; This study describes a potential effect of topical gentian violet on cutaneous infection and nasal carriage with methicillin-resistant Staphylococcus aureus (MRSA) . 0.5% gentian violet was used in 28 cases of skin lesions once a day, while a 0.3% solution was applied on the nasal vestibules of nine cases twice a day . The period for eradication in the 28 skin cases was 9.1 +/- 6.0 days . It was 15.3 +/- 9.0 days for the nine nasal lesions . The minimal inhibitory concentration (MIC) of gentian violet against MRSA from the four isolated strains was 0.0225 +/- 0.0096 microg/mL . No adverse reactions occurred throughout the study . It is suggested that gentian violet may be potentially effective against MRSA. Immunology, 2001 Jan, 102(1), 103 - 13 Mice lacking tartrate-resistant acid phosphatase (Acp 5) have disordered macrophage inflammatory responses and reduced clearance of the pathogen, Staphylococcus aureus; Bune AJ et al.; Tartrate-resistant acid phosphatase (TRAP) is a lysosomal di-iron protein of mononuclear phagocytes and osteoclasts . Hitherto, no role for the enzyme in immunity has been identified; however, knockout mice lacking TRAP have a skeletal phenotype caused by an intrinsic osteoclast defect . To investigate a putative function for TRAP in macrophages (Mphi), we investigated proinflammatory responses and systemic microbial clearance in knockout mice compared with age- and gender-matched congenic wild-type mice . Phorbol 12-myristate 13-acetate (PMA)-stimulated and interferon-gamma (IFN-gamma)-induced superoxide formation was enhanced in peritoneal Mphi lacking TRAP; nitrite production in response to stimulation with lipopolysaccharide (LPS) and IFN-gamma was also increased . In addition, secretion of the proinflammatory cytokines, tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta and IL-12, was significantly greater in TRAP-deficient Mphi when stimulated with LPS, with or without addition of either TNF-alpha or IFN-gamma . The activity of tartrate-sensitive (lysosomal) acid phosphatase was increased in Mphi from the knockout mice but activities of the lysosomal hydrolases N-acetyl beta-glucosaminidase and acid beta-glucuronidase were unchanged, indicating selective activation of compensatory acid phosphatase activity . Evidence of impaired Mphi function in vivo was obtained in TRAP knockout mice, which showed delayed clearance of the microbial pathogen, Staphylococcus aureus, after sublethal intraperitoneal inoculation . After microbial challenge, peritoneal exudates obtained from TRAP knockout mice had a reduced population of Mphi . As peritoneal Mphi and neutrophils lacking TRAP were able to phagocytose and kill S . aureus normally in vitro, TRAP may directly or indirectly influence recruitment of Mphi to sites of microbial invasion . Our study shows that TRAP participates in the inflammatory response of the Mphi and influences effector signalling pathways in innate immunity. Clin Microbiol Infect, 2000 Sep, 6(9), 483 - 9 MIC determination of fusidic acid and of ciprofloxacin using multidisk diffusion tests; Kronvall G; OBJECTIVE: To investigate the possibility of estimating the MICs of fusidic acid and ciprofloxacin for bacterial isolates using series of antibiotic disk concentrations in diffusion tests, so-called M-tests . METHODS: Thirty Staphylococcus aureus and S . epidermidis strains were tested for fusidic acid susceptibility . Sixty-one clinical isolates of eight bacterial species were tested for ciprofloxacin susceptibility . Disk diffusion was standardized according to the Swedish reference group for antibiotics (SRGA) . For fusidic acid, a series of disks (1.5, 5.0, 15, 50 and 150 microg) was used . Ciprofloxacin was applied in four different diffusion sources (1, 3, 10 and 30 microg) on a single strip, the M-strip, and used . True MIC values were determined using the standardized agar dilution method according to the SRGA . Single-strain regression analysis (SRA) was employed to calculate critical concentration equivalents (Qzero) . RESULTS: Fusidic acid and ciprofloxacin critical concentrations were determined for the bacterial isolates . The mean conversion factors for Qzero to yield the true MIC were 2.06 (range 0.34-8.9) for fusidic acid and 2.05 (range 0.37-8.1) for ciprofloxacin . There was a correlation between true MIC values (all MICs expressed as 2 log + 9) and the calculated MIC values (Qzero x conversion factor) for both fusidic acid (R = 0.9822) and ciprofloxacin (R = 0.9696) . CONCLUSIONS: MIC values of clinical isolates can be estimated using SRA calculations on zone measurements in disk tests with several concentrations of the antibiotic in diffusion sources. Clin Microbiol Infect, 2000 May, 6(5), 239 - 45 Clonal dissemination of epidemic methicillin-resistant Staphylococcus aureus in Belgium and neighboring countries; Deplano A et al.; OBJECTIVES: To determine the diversity of pulsed-field gel electrophoresis (PFGE) types among epidemic strains of methicillin-resistant Staphylococcus aureus (MRSA) recovered in Belgium, France, Germany and The Netherlands over the period 1981-94 . METHODS: MRSA strains collected in a multicenter survey in Belgium (n = 171) and from reference laboratories in neighboring countries (n = 102) were characterized by PFGE analysis using the SmaI enzyme . RESULTS: In total, 32 PFGE types were found . Epidemic PFGE type 1, first recognized in 1984, accounted for 82% of Belgian strains (87% of hospitals) and 51% of European MRSA strains . Four other internationally epidemic PFGE types (types 8, 10, 11 and 12) were less widely disseminated and more recently detected (1991-94), each recovered from two or three countries . International spread of two PFGE types was linked to transfer of colonized patients to Dutch hospitals from another country where this type was frequently recovered . CONCLUSIONS: Genotypic analysis indicated widespread distribution of several outbreak-associated MRSA strains over large European regions, which was in some instances related to interhospital patient transfer . These findings underscore the need for standardized international surveillance and control of MRSA transmission between healthcare institutions across Europe. Vet Microbiol, 2001 Jan 26, 78(2), 183 - 91 Effect of slime on adherence of Staphylococcus aureus isolated from bovine and ovine mastitis; Aguilar B et al.; The interactions between slime, Staphylococcus aureus and ovine mammary gland epithelial cells (MGEC) were studied in vitro . Suspensions of radiolabelled bacteria incubated with slime significantly increased the ability of S . aureus strains to adhere to a filter . When suspensions of radiolabelled bacteria were incubated with MGEC treated with trypsin, the ability of slime to improve S . aureus adherence was also shown, indicating that it was not dependent on cell membrane proteins . The interaction of radiolabelled bacteria with slime prior to the adherence test with MGEC demonstrated that the adherence process requires the interaction between slime and bacteria . This interaction is inhibited by anti-slime antibodies . This study provides evidence that a specific interaction between bacteria coated with slime and MGEC could be a critical part of mammary gland infection. Biochem Biophys Res Commun, 2001 Feb 2, 280(4), 1028 - 35 Molecular cloning and functional characterization of a human scavenger receptor with C-type lectin (SRCL), a novel member of a scavenger receptor family; Nakamura K et al.; Using a human placenta cDNA library, we cloned a novel member belonging to the scavenger receptor family . Complementary DNA of this clone encodes a type II transmembranous glycoprotein containing a collagen-like domain, which are typical structural characteristics of scavenger receptor class A . This protein also contains a C-type lectin/carbohydrate recognition domain (C-type CRD) located at the C-terminus . We designated this as Scavenger Receptor with C-type Lectin (SRCL) type I . We also isolated human SRCL type II, which lacks the C-type CRD . Northern blot analysis revealed that hSRCL type I and type II mRNAs are abundantly expressed in adult human tissues . When hSRCL type I and type II were expressed in CHO-K1 cells, they were localized in the plasma membrane forming clusters on the surface . Ligand-binding studies of CHO-K1 cells expressing hSRCL type I and type II demonstrated their specific binding capacity in Escherichia coli and Staphylococcus aureus . These results indicate that hSRCL is a novel bacteria-binding receptor containing a C-type CRD and this receptor may play an important role in host defense . Protein Expr Purif, 2001 Feb, 21(1), 176 - 82 Recombinant human insulin . VIII . Isolation of fusion protein--S-sulfonate, biotechnological precursor of human insulin, from the biomass of transformed Escherichia coli cells; Tikhonov RV et al.; Various methods have been investigated for the isolation and purification of fusion proteins of precursors of human insulin in the form of S-sulfonates, from the biomass of transformed Escherichia coli cells . Fusion proteins were prepared with different sizes and structures of the leader peptide and the poly-His position (inserted for purification by metal chelate affinity chromatography) . The fusion proteins contained an IgG-binding B domain of protein A from Staphylococcus aureus at the N-terminus and an Arg residue between the leader peptide of the molecule and the proinsulin sequence, for trypsin cleavage of the leader peptide . Six residues of Cys in proinsulin allow the chemical modification of the protein as a (Cys-S-SO(-)(3))(6) derivative (S-sulfonate), which increases its polyelectrolytic properties and improves the efficiency of its isolation . Various methods of oxidative sulfitolysis were compared with catalysis by sodium tetrathionate or cystine and Cu2+ or Ni2+ ions . An optimum scheme for the isolation and purification of S-sulfonated fusion proteins was developed by the combination of metal-chelating affinity and ion-exchange chromatography . Highly purified (95%) S-sulfonated fusion protein was recovered which was 85% of the fusion protein contained in the biomass of E . coli cells . Folding of fusion protein S-sulfonate occurred with high yield (up to 90-95%) . We found that the fusion protein-S-sulfonate has proinsulin-like secondary structure.This structure causes highly efficient fusion protein folding . J Hosp Infect, 2001 Jan, 47(1), 9 - 18 Screening for methicillin-resistant Staphylococcus aureus in the endemic hospital: what have we learned? Rubinovitch B, Pittet D. Control of methicillin-resistant Staphylococcus aureus (MRSA) still generates controversy among infection control practitioners . Opponents claim that once MRSA becomes endemic in an institution, control efforts are no longer justified . This review examines the usefulness, feasibility and cost-effectiveness of control programmes in acute-care hospitals where eradication of MRSA has either failed or has never been attempted; hence, the pathogen has become endemic . High endemicity is associated with increased hospital-acquired infection rates, increased use of glycopeptides and subsequent risk of emergence of antibiotic-resistant Gram-positive bacteria, and additional healthcare costs . Thus, MRSA control has many advantages . Indeed, in many institutions the actual benefit of containment efforts was manifested through the resultant decrease in the incidence of hospital-acquired MRSA infections . Successful programmes are based on an early identification of the MRSA reservoir and prompt implementation of contact precautions . The most efficacious strategy to detect occult MRSA carriage is via the screening of high-risk patients on admission to the hospital which has proven to be cost-effective in varied acute-care endemic settings . J Biol Chem, 2001 Jan 26, 276(4), 2658 - 67 Regulation of Staphylococcus aureus pathogenesis via target of RNAIII-activating Protein (TRAP); Balaban N et al.; Staphylococcus aureus can cause disease through the production of toxins . Toxin production is autoinduced by the protein RNAIII-activating protein (RAP) and by the autoinducing peptide (AIP), and is inhibited by RNAIII-inhibiting peptide (RIP) and by inhibitory AIPs . RAP has been shown to be a useful vaccine target site, and RIP and inhibitory AIPs as therapeutic molecules to prevent and suppress S . aureus infections . Development of therapeutic strategies based on these molecules has been hindered by a lack of knowledge of the molecular mechanisms by which they activate or inhibit virulence . Here, we show that RAP specifically induces the phosphorylation of a novel 21-kDa protein, whereas RIP inhibits its phosphorylation . This protein was termed target of RAP (TRAP) . The synthesis of the virulence regulatory molecule, RNAIII, is not activated by RAP in the trap mutant strain, suggesting that RAP activates RNAIII synthesis via TRAP . Phosphoamino acid analysis shows that TRAP is histidine-phosphorylated, suggesting that TRAP may be a sensor of RAP . AIPs up-regulate the synthesis of RNAIII also in trap mutant strains, suggesting that TRAP and AIPs activate RNAIII synthesis via distinct signal transduction pathways . Furthermore, TRAP phosphorylation is down-regulated in the presence of AIP, suggesting that a network of signal transduction pathways regulate S . aureus pathogenesis. Infect Immun, 2001 Feb, 69(2), 917 - 23 Staphylococcus aureus Cap5O has UDP-ManNAc dehydrogenase activity and is essential for capsule expression; Portoles M et al.; The Staphylococcus aureus serotype 5 capsular polysaccharide (CP5) has a repeating unit composed of (-->4)-3-O-acetyl-beta-D-ManNAcA-(1-->4)-alpha-L-FucNAc (1-->3)-beta-D-FucNAc-(1-->)(n) . Sixteen chromosomal genes (cap5A through cap5P) are involved in the synthesis of CP5 . We recently demonstrated that Cap5P, a 2-epimerase, catalyzes the conversion of UDP-N-acetyl glucosamine (UDP-GlcNAc) to UDP-N-acetylmannosamine (UDP-ManNAc) . In this study, we show that UDP-ManNAc is oxidized to UDP-N-acetylmannosaminuronic acid (UDP-ManNAcA) by a UDP-ManNAc dehydrogenase encoded by S . aureus cap5O . We expressed Cap5O in Escherichia coli and purified the recombinant protein . The UDP-ManNAc dehydrogenase activity of purified Cap5O was assessed by incubating Cap5P and UDP-GlcNAc (to produce UDP-ManNAc), together with Cap5O, NAD(+), and a reducing agent . Enzymatic activity was quantitated indirectly by measuring the increase in absorbance at 340 nm resulting from NADH formation . The product of the reaction was confirmed as UDP-ManNAcA by gas chromatography-mass spectroscopy . A cap5O mutation, created by deletion of 727 bp in the 5' end of the gene, was introduced by allelic replacement into S . aureus Reynolds, rendering it CP5 negative . Mice inoculated intravenously or subcutaneously with the wild-type strain Reynolds had greater numbers of S . aureus recovered from their kidneys (P = 0.019) or their subcutaneous abscesses (P = 0.0018), respectively, than did animals inoculated with the cap5O mutant . The results of this study indicate that S . aureus cap5O is essential for capsule production and that capsule promotes staphylococcal virulence in mouse models of abscess formation. Infect Immun, 2001 Feb, 69(2), 885 - 96 Characterization of sarR, a modulator of sar expression in Staphylococcus aureus; Manna A et al.; The expression of virulence determinants in Staphylococcus aureus is controlled by global regulatory loci (e.g., sar and agr) . The sar locus is composed of three overlapping transcripts (sar P1, P3, and P2 transcripts from P1, P3, and P2 promoters, respectively), all encoding the 372-bp sarA gene . The level of SarA, the major regulatory protein, is partially controlled by the differential activation of sar promoters . We previously partially purified a approximately 12 kDa protein with a DNA-specific column containing a sar P2 promoter fragment . In this study, the putative gene, designated sarR, was identified and found to encode a 13.6-kDa protein with homology to SarA . Transcriptional and immunoblot studies revealed the sarR gene to be expressed in other staphylococcal strains . Recombinant SarR protein bound sar P1, P2, and P3 promoter fragments in gel shift and footprinting assays . A sarR mutant expressed a higher level of P1 transcript than the parent, as confirmed by promoter green fluorescent protein fusion assays . As the P1 transcript is the predominant sar transcript, we confirmed that the sarR mutant expressed more SarA than the parental strain . We thus proposed that SarR is a regulatory protein that binds to the sar promoters to down-regulate P1 transcription and the ensuing SarA protein expression. Arch Dis Child, 2001 Feb, 84(2), 160 - 2 Methicillin resistant Staphylococcus aureus (MRSA) infection in cystic fibrosis; Miall LS et al.; BACKGROUND: Methicillin resistant Staphylococcus aureus (MRSA) infection is increasingly found in patients with cystic fibrosis (CF) . AIMS: To determine whether MRSA infection has a deleterious effect on the clinical status of children with CF . METHODS: Children with MRSA in respiratory cultures during a seven year period were identified and compared with controls matched for age, sex, and respiratory function . Respiratory function tests, anthropometric data, Shwachman-Kulczycki score, Northern chest x ray score, intravenous and nebulised antibiotic therapy, and steroid therapy were compared one year before and one year after MRSA infection . RESULTS: From a clinic population of 300, 10 children had positive sputum or cough swab cultures for MRSA . Prevalence rose from 0 in 1992-1994 to 7 in 1998 . Eighteen controls were identified . Children with MRSA showed significant worsening of height standard deviation scores and required twice as many courses of intravenous antibiotics as controls after one year . They had significantly worse chest x ray scores at the time of the first MRSA isolate and one year later, but showed no increase in the rate of decline in chest x ray appearance . There was a trend towards lower FEV(1) and FEF(25-75) in children with MRSA . There were no significant differences between the two groups with respect to change in weight, body mass index, or Shwachman score . There was no significant difference in prior use of steroids or nebulised antibiotics . CONCLUSION: MRSA infection in children with CF does not significantly affect respiratory function, but may have an adverse effect on growth . Children with MRSA require significantly more courses of intravenous antibiotics and have a worse chest x ray appearance than controls. Antimicrob Agents Chemother, 2001 Feb, 45(2), 454 - 9 Bactericidal activities of two daptomycin regimens against clinical strains of glycopeptide intermediate-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecium, and methicillin-resistant Staphylococcus aureus isolates in an in vitro pharmacodynamic model with simulated endocardial vegetations; Akins RL et al.; Daptomycin is an investigational lipopeptide antibiotic active against gram-positive organisms . The mechanism of action is unique, resulting in interference with cell membrane transport . The bactericidal activity of daptomycin was evaluated against glycopeptide-intermediate susceptible Staphylococcus aureus (GISA), vancomycin-resistant Enterococcus faecium (VREF), and methicillin-resistant S . aureus (MRSA) in an in vitro infection model with simulated endocardial vegetations . Simulated regimens of daptomycin at 6 mg/kg/day (D6) and 10 mg/kg/day (D10) were utilized . MICs and MBCs for daptomycin were determined in the absence and in the presence of albumin with the following results (MIC/MBC): for GISA-992, 0.5/1.0 and 16/16; for VREF-590, 2.0/2.0 and 32/32; and for MRSA-494, 0.25/0.25 and 1.0/4.0 microg/ml, respectively . During the first 8 h daptomycin significantly reduced the inoculum for all organisms . Daptomycin at 6 mg/kg/day and 10 mg/kg/day had log(10) CFU/g reductions of 5 and 6, 3.4 and 5, and 6.4 and 6.5 by 8 h for GISA-992, VREF-590, and MRSA-494, respectively . Against both GISA-992 and VREF-590, the D10 regimen achieved the limit of detection at 72 h, with D6 regimens showing slight regrowth . A concentration-dependent killing effect was noted to occur, with daptomycin demonstrating a more rapid and greater kill from the D10 versus the D6 regimen . The results of this study suggest that daptomycin demonstrates significant (P < 0.05) activity against gram-positive organisms in a simulated sequestered infection site. Antimicrob Agents Chemother, 2001 Feb, 45(2), 407 - 12 The AbcA transporter of Staphylococcus aureus affects cell autolysis; Schrader-Fischer G et al.; Increased production of penicillin-binding protein PBP 4 is known to increase peptidoglycan cross-linking and contributes to methicillin resistance in Staphylococcus aureus . The pbp4 gene shares a 400-nucleotide intercistronic region with the divergently transcribed abcA gene, encoding an ATP-binding cassette transporter of unknown function . Our study revealed that methicillin stimulated abcA transcription but had no effects on pbp4 transcription . Analysis of abcA expression in mutants defective for global regulators showed that abcA is under the control of agr . Insertional inactivation of abcA by an erythromycin resistance determinant did not influence pbp4 transcription, nor did it alter resistance to methicillin and other cell wall-directed antibiotics . However, abcA mutants showed spontaneous partial lysis on plates containing subinhibitory concentrations of methicillin due to increased spontaneous autolysis . Since the autolytic zymograms of cell extracts were identical in mutants and parental strains, we postulate an indirect role of AbcA in control of autolytic activities and in protection of the cells against methicillin. Proc Natl Acad Sci U S A, 2001 Jan 30, 98(3), 932 - 7 Epub 2001 Jan 23. An unusual isopentenyl diphosphate isomerase found in the mevalonate pathway gene cluster from Streptomyces sp . strain CL190; Kaneda K et al.; A gene cluster encoding five enzymes of the mevalonate pathway had been cloned from Streptomyces sp . strain CL190 . This gene cluster contained an additional ORF, orfD, encoding an unknown protein that was detected in some archaebacteria and some Gram-positive bacteria including Staphylococcus aureus . The recombinant product of orfD was purified as a soluble protein and characterized . The molecular mass of the enzyme was estimated to be 37 kDa by SDS-polyacrylamide gel electrophoresis and 155 kDa by gel filtration chromatography, suggesting that the enzyme is most likely to be a tetramer . The purified enzyme contained flavin mononucleotide (FMN) with the amount per tetramer being 1.4 to 1.6 mol/mol . The enzyme catalyzed the isomerization of isopentenyl diphosphate (IPP) to produce dimethylallyl diphosphate (DMAPP) in the presence of both FMN and NADPH . The Escherichia coli plasmid expressing orfD could complement the disrupted IPP isomerase gene in E . coli . These results indicate that orfD encodes an unusual IPP isomerase showing no sequence similarity to those of IPP isomerases identified to date . Based on the difference in enzymatic properties, we classify the IPP isomerases into two types: Type 2 for FMN- and NAD(P)H-dependent enzymes, and type 1 for the others . In view of the critical role of this isomerase in S . aureus and of the different enzymatic properties of mammalian (type 1) and S . aureus (type 2) isomerases, this unusual enzyme is considered to be a suitable molecular target for the screening of antibacterial drugs specific to S . aureus. J Clin Microbiol, 2001 Feb, 39(2), 779 - 81 Nosocomial spread of an unusual methicillin-resistant Staphylococcus aureus clone that is sensitive to all non-beta-lactam antibiotics, including tobramycin; Pournaras S et al.; Between January and December 1999, in Hippokration General Hospital, Thessaloniki, Greece, a large proportion of the methicillin-resistant Staphylococcus aureus isolates (34.4%) exhibited susceptibility to virtually all alternative non-beta-lactam antibiotics, including tobramycin . Twenty-five of them were selected randomly for further testing; all belonged to a unique genotype and were characterized as heterogeneously resistant to oxacillin . The aadD gene, encoding tobramycin resistance, failed to be amplified in all cases, indicating absence of the gene or the entire plasmid pUB110 from the mec DNA. J Clin Microbiol, 2001 Feb, 39(2), 728 - 9 Breast milk transmission of a Panton-Valentine leukocidin-producing Staphylococcus aureus strain causing infantile pneumonia; Le Thomas I et al.; We report on a 38-day-old infant who developed pleuropneumonia due to a Staphylococcus aureus strain responsible for familial furunculosis, which was acquired by maternal breast-feeding . All isolates from the infant and parents were genetically related by randomly amplified polymorphic DNA analysis and produced Panton-Valentine leukocidin. J Clin Microbiol, 2001 Feb, 39(2), 591 - 5 First report of methicillin-resistant Staphylococcus aureus with reduced susceptibility to vancomycin in Thailand; Trakulsomboon S et al.; To investigate whether there are methicillin-resistant Staphylococcus aureus (MRSA) strains with reduced susceptibility to vancomycin in Thailand, a total of 155 MRSA strains isolated from patients hospitalized between 1988 and 1999 in university hospitals in Thailand were tested for glycopeptide susceptibility . All the strains were classified as susceptible to vancomycin and teicoplanin when judged by NCCLS criteria for glycopeptide susceptibility using the agar dilution MIC determination . Vancomycin MICs at which 50 and 90% of the isolates tested were inhibited (MIC50 and MIC(90), respectively) were 0.5 and 1 microg/ml, respectively, with a range of 0.25 to 2 microg/ml . For teicoplanin, MIC50 and MIC90 were 2 microg/ml, with a range of 0.5 to 4 microg/ml . However, one-point population analysis identified three MRSA strains, MR135, MR187, and MR209, which contained subpopulations of cells that could grow in 4 microg of vancomycin per ml . The proportions of the subpopulations were 2 x 10(-4), 1.5 x 10(-6), and 4 x 10(-7), respectively . The subsequent performance of a complete population analysis and testing for the emergence of mutants with reduced susceptibility to vancomycin (MIC > or = 8 microg/ml) confirmed that these strains were heterogeneously resistant to vancomycin . Two of these strains caused infection that was refractory to vancomycin therapy . Pulsed-field gel electrophoresis showed that the two strains had identical SmaI macrorestriction patterns and that they were one of the common types of MRSA isolated in the hospital . This is the first report of heterogeneous resistance to vancomycin in Thailand and an early warning for the possible emergence of vancomycin resistance in S . aureus in Southeast Asia. J Clin Microbiol, 2001 Feb, 39(2), 574 - 80 Comparison of DNA sequencing of the protein A gene polymorphic region with other molecular typing techniques for typing two epidemiologically diverse collections of methicillin-resistant Staphylococcus aureus; Oliveira DC et al.; The aim of this study was to compare the recently developed typing approach for methicillin-resistant Staphylococcus aureus (MRSA) based on the DNA sequencing of the protein A gene polymorphic region (spaA typing) with a combination of three well-established molecular typing techniques: ClaI-mecA vicinity polymorphisms, ClaI-Tn554 insertion patterns, and SmaI pulsed-field gel electrophoresis (PFGE) profiles . In order to evaluate the applicability of this typing technique in different types of studies, two groups of MRSA clinical isolates were analyzed: a collection of 185 MRSA isolates circulating in Hungary recovered from 17 hospitals in seven cities during a 3-year period (1994 through 1996), and a selection of 53 MRSA strains isolated in a single hospital in Hungary between 1997 and 1998 . The 238 MRSA clinical strains from Hungary were first classified in clonal types (defined as ClaI-mecA::ClaI-Tn554::SmaI-PFGE patterns), and 65 of the 238 strains, representing major MRSA clones and some sporadic clones, were further analyzed by spaA typing . Our results showed that the lineages most recently introduced in the hospital setting showed little variability in spaA types, whereas the MRSA clones circulating for a longer period of time and spread among several hospitals showed a higher degree of variability . The implementation of the spaA typing method was straightforward, and the results obtained were reproducible, unambiguous, and easily interpreted . This method seems to be adequate for outbreak investigations but should be complemented with other techniques in long-term surveillance or in studies comparing distant clonal lineages. J Antimicrob Chemother, 2001 Feb, 47(2), 187 - 9 Evaluation of the Mastalex latex agglutination test for methicillin resistance in Staphylococcus aureus grown on different screening media; Brown DF et al.; The Mastalex MRSA latex agglutination method was evaluated with 52 methicillin-resistant and 27 methicillin-susceptible strains of Staphylococcus aureus grown on various media . All tests were correct with colonies grown on blood agar with or without oxacillin 2 mg/L . Tests on colonies grown on mannitol salt agar were less reliable, although addition of oxacillin 2 mg/L improved performance . One of the 26 MRSA which grew on Baird-Parker medium with 8 mg/L ciprofloxacin gave a false-negative result . Agglutination was faster when strains were grown on media with oxacillin . The method would be particularly useful for urgent confirmation of resistance. J Antimicrob Chemother, 2001 Feb, 47(2), 157 - 61 Molecular diversity of quinolone resistance in genetically related clinical isolates of Staphylococcus aureus and susceptibility to newer quinolones; Guirao GY et al.; The genes encoding topoisomerases (gyrA and grlA) and the norA promoter of 100 fluoroquinolone-susceptible and -resistant Staphylococcus aureus clinical isolates obtained in two geographically distant hospitals were analysed . The relationship between mutations found and the susceptibility to newer quinolones was determined . Thirty-nine strains were grouped in seven clones by pulsed-field gel electrophoresis (PFGE) . The remaining 61 strains were classified as unrelated strains . In three clones, all strains showed the same grlA-gyrA-norA mutation profiles . Strains in the rest of the groups showed different mutation profiles, even though PFGE indicated that they possessed genetically similar populations . One cluster showed a high level of diversity; five different mutation profiles were detected in the six isolates belonging to this pattern . Two isolates had a Glu84 to Lys mutation in grlA and another isolate had this mutation combined with a Ser84 to Leu mutation in gyrA . Combination of a Ser80 to Phe mutation in grlA and a Ser84 to Leu in gyrA was found in the two other isolates . One of these also had a thymine to a guanine transversion at a position 89 nucleotides upstream of the norA start codon in the norA promoter . These results show that fluoroquinolone resistance in clinical S . aureus strains does not necessarily result from the spread of resistant clones . Fluoroquinolone resistance may develop independently in strains belonging to the same PFGE pattern by accumulation of different mutations over a quinolone-susceptible ancestor wild type or single grlA mutant. J Antimicrob Chemother, 2001 Feb, 47(2), 153 - 6 Differential effect of rpoB mutations on antibacterial activities of rifampicin and KRM-1648 against Staphylococcus aureus; Wichelhaus T et al.; The in vitro antibacterial activities of the rifamycin derivatives rifampicin and KRM-1648 against 150 Staphylococcus aureus isolates were determined . The MICs of rifampicin and KRM-1648 for 90% of rifampicin-susceptible S . aureus isolates (n = 100) were 0.016 and 0.001 mg/L, respectively . In rifampicin-resistant S . aureus isolates (n = 50), different levels of resistance to rifamycins were associated with mutations at different sites in rpoB . Mutations at some sites were associated with high-level resistance to both rifamycins, while certain mutations were associated with the activity of KRM-1648 being < or = 100-fold better than that of rifampicin. Int Immunol, 2001 Feb, 13(2), 157 - 66 Comparison of thymocyte development and cytokine production in CD7-deficient, CD28-deficient and CD7/CD28 double-deficient mice; Heinly CS et al.; CD7 and CD28 are Ig superfamily molecules expressed on thymocytes and mature T cells that share common signaling 0mechanisms and are co-mitogens for T cell activation . CD7-deficient mice are resistant to lipopolysaccharide (LPS)-induced shock syndrome, and have diminished in vivo LPS-triggered IFN-gamma and tumor necrosis factor (TNF)-alpha production . CD28-deficient mice have decreased serum Ig levels, defective IgG isotype switching, decreased T cell IL-2 production and are resistant to Staphylococcus aureus enterotoxin-induced shock . To determine synergistic roles CD7 and CD28 might play in thymocyte development and function, we have generated and characterized CD7/CD28 double-deficient mice . CD7/CD28-deficient mice were healthy, reproduced normally, had normal numbers of thymocyte subsets and had normal thymus histology . Anti-CD3 mAb induced similar levels of apoptosis in CD7-deficient, CD28-deficient and CD7/CD28 double-deficient thymocytes as in control C57BL/6 mice (P = NS) . Similarly, thymocyte viability, apoptosis and necrosis following ionomycin or dexamethasone treatment were the same in control, CD7-deficient, CD28-deficient and CD7/CD28-deficient mice . CD28-deficient and CD7/CD28-deficient thymocytes had decreased {3H}thymidine incorporation responses to concanavalin A (Con A) stimulation compared to control mice (P < or = 0.01 and P < or = 0.05 respectively) . CD7/CD28 double-deficient mice had significantly reduced numbers of B7-1/B7-2 double-positive cells compared to freshly isolated wild-type, CD7-deficient and CD28-deficient thymocytes . Con A-stimulated CD4/CD8 double-negative (DN) thymocytes from CD7/CD28 double-deficient mice expressed significantly lower levels of CD25 when compared to CD4/CD8 DN thymocytes from wild-type, CD7-deficient and CD28-deficient mice (P < 0.05) . Anti-CD3-triggered CD7/CD28-deficient thymocytes also had decreased IFN-gamma and TNF-alpha production compared to C57BL/6 control, CD7-deficient and CD28-deficient mice (P < or = 0.05) . Thus, CD7 and CD28 deficiencies combined to produce abnormalities in the absolute number of B7-1/B7-2-expressing cells in the thymus, thymocyte IL-2 receptor expression and CD3-triggered cytokine production. Appl Environ Microbiol, 2001 Feb, 67(2), 1001 - 3 Molecular characterization of the iron-hydroxamate uptake system in Staphylococcus aureus; Cabrera G et al.; To investigate iron uptake, a chromosomal locus containing three consecutive open reading frames, designated fhuC, fhuB, and fhuD, was identified in Staphylococcus aureus . Whereas the fhuC gene encodes an ATP-binding protein, fhuB and fhuD code for ferrichrome permeases and thus resemble an ATP-binding cassette transporter . A fhuB knockout mutant showed impaired uptake of iron bound to the siderophores but not of ferric chloride, suggesting that this operon is specific for siderophore-mediated iron uptake. J Exp Med, 2001 Feb 5, 193(3), 393 - 7 Structure-function relationship of cytokine induction by lipoteichoic acid from Staphylococcus aureus; Morath S et al.; Lipoteichoic acids (LTAs) have been proposed as putative Gram-positive immunostimulatory counterparts to Gram-negative lipopolysaccharides . However, LTA from Staphylococcus aureus, the clinically most frequent Gram-positive pathogen, was inactive after purification . Here, a novel isolation procedure to prepare pure (>99%) biologically active LTA, allowing the first structural analysis by nuclear magnetic resonance and mass spectrometry, is described . A comparison with LTA purified by standard techniques revealed that alanine substituents are lost during standard purification, resulting in attenuated cytokine induction activity . In line with this finding, hydrolysis of alanine substituents of active LTA decimated cytokine induction . LTA represents a major immunostimulatory component of S . aureus. J Immunol, 2000 Apr 15, 164(8), 4097 - 104 CD134L engagement enhances human B cell Ig production: CD154/CD40, CD70/CD27, and CD134/CD134L interactions coordinately regulate T cell-dependent B cell responses; Morimoto S et al.; CD134 is a member of the TNFR family expressed on activated T cells, whose ligand, CD134L, is found preferentially on activated B cells . We have previously reported that the CD70/CD27 interaction may be more important in the induction of plasma cell differentiation after the expansion phase induced by the CD154/CD40 interaction has occurred . When CD134-transfected cells were added to PBMCs stimulated with pokeweed mitogen, IgG production was enhanced in a dose-dependent fashion . Addition of CD134-transfected cells to B cells stimulated with Staphylococcus aureus Cowan I strain/IL-2 resulted in little if any enhancement of B cell IgG production and proliferation . We found that while CD134-transfected cells induced no IgG production by themselves, it greatly enhanced IgG production in the presence of CD40 stimulation or T cell cytokines such as IL-4 and IL-10 . The addition of CD134-transfected cells showed only a slight increase in the number of plasma cells compared with that in the culture without them, indicating that an increased Ig production rate per cell is responsible for the observed enhancing effect of CD134L engagement rather than increase in plasma cell generation . These results strongly suggest different and sequential roles of the TNF/TNFR family molecules in human T cell-dependent B cell responses through cell-cell contacts and the cytokine network. Biochem Biophys Res Commun, 2000 Apr 13, 270(2), 387 - 92 A conserved residue at the extreme C-terminus of FtsZ is critical for the FtsA-FtsZ interaction in Staphylococcus aureus; Yan K et al.; FtsZ, a tubulin-like protein with GTPase activity, and FtsA, an actin-like protein with ATPase activity, are two proteins known to play critical roles in the later stages of the bacterial cell cycle . It is well documented that FtsA-FtsZ co-localization at the septum of dividing bacteria is essential for successful cell division in E . coli . We have validated and characterized this interaction by co-expressing FtsA and FtsZ, from both E . coli and S . aureus, in the yeast two-hybrid system . We demonstrate for the first time a specific association between S . aureus FtsA and FtsZ proteins and self interaction of S . aureus FtsZ . These observations are consistent with the conserved role of FtsA and FtsZ in bacterial cell division . Using deletion mutagenesis, we have shown that a highly conserved motif as small as 10 residues in the extreme C-terminus of S . aureus FtsZ is critical for the interaction with FtsA . Further dissection of this motif by alanine scanning mutagenesis showed that Phe376 likely plays a major role in the FtsA-FtsZ interaction . This work has important implications for the design of antagonists and agonists of the FtsA-FtsZ interaction as such agents could provide a novel approach for tackling multi-resistant Gram positive pathogens . Rev Assoc Med Bras, 1999 Oct-Dec, 45(4), 371 - 4 {Bacterial endocarditis as a complication of neonatal sepsis: case report}; Krebs VL et al.; The authors reported on a 11 day-old child, admitted in Neonatal Intensive Care Unit for multiple congenital malformations, who had sepsis and bacterial endocarditis . Among the risk factors for endocarditis were outstanding: the central venous catheterism, hemoculture with growth of Staphylococcus aureus and mechanical ventilation . The diagnosis was made in the 61st day after admission owing to the presence of persistent fever and appearance of systolic murmur . The echocardiogram revealed a thrombus in the right atrium measuring 1.9 x 0.7 mm . Antibiotic therapy and surgical resection being performed, with clinical improvement . On the 125st day after admission the patient died owing sepsis and cerebral abscess . At necropsy, heart malformations were not observed . The authors concluded to be very important the knowledge of the potential risks of invasive procedures currently used to care for critically ill newborns . The clinical suspicion of endocarditis should be considered in all neonates with sepsis and receiving intensive care for long time. J Biol Chem, 2001 Apr 13, 276(15), 12324 - 30 Epub 2001 Jan 04. Major photoaffinity drug binding sites in multidrug resistance protein 1 (MRP1) are within transmembrane domains 10-11 and 16-17; Daoud R et al.; MRP1 is an ABC (or ATP binding cassette) membrane transport protein shown to confer resistance to structurally dissimilar drugs . Studies of MRP1 topology suggested the presence of a hydrophobic N-domain with five potential membrane-spanning domains linked to an MDR1-like core (MSD1-NBD1-L1-MSD2-NBD2) by an intracellular linker domain (L0) . MRP1-mediated multidrug resistance is thought to be due to enhanced drug efflux . However, little is known about MRP1-drug interaction and its drug binding site(s) . We previously developed several photoreactive probes to study MRP1-drug interactions . In this report, we have used eight MRP1-HA variants that were modified to have hemagglutinin A (HA) epitopes inserted at different sites in MRP1 sequence . Exhaustive in-gel digestion of all IAARh123 photoaffinity-labeled MRP1-HA variants revealed the same profile of photolabeled peptides as seen for wild type MRP1 . Photolabeling of the different MRP1-HA variants followed by digestion with increasing concentrations of trypsin or Staphylococcus aureus V8 protease (1:800 to 1:5 w/w) and immunoprecipitation with anti-HA mAb identified two small photolabeled peptides ( approximately 6-7 kDa) from MRP1-HA(574) and MRP1-HA(1222) . Based on the location of the HA epitopes in the latter variants together with molecular masses of the two peptides, the photolabeled amino acid residues were localized to MRP1 sequences encoding transmembranes 10 and 11 of MSD1 (Ser(542)-Arg(593)) and transmembranes 16 and 17 of MSD2 (Cys(1205)-Glu(1253)) . Interestingly, the same sequences in MRP1 were also photolabeled with a structurally different photoreactive drug, IACI, confirming the significance of transmembranes 10, 11, 16 and 17 in MRP1 drug binding . Taken together, the results in this study provide the first delineation of the drug binding site(s) of MRP1 . Furthermore, our findings suggest the presence of common drug binding site(s) for structurally dissimilar drugs. J Formos Med Assoc, 2000 Dec, 99(12), 942 - 4 Primary staphylococcal infection and toxic shock syndrome diagnosed by polymerase chain reaction; Tsai YG et al.; Primary staphylococcal pneumonia complicated with toxic shock syndrome (TSS) is relatively uncommon in children . Staphylococcus aureus exotoxins are thought to function as superantigens, and seem to promote disease manifestations . The identification of staphylococcal toxin genes by polymerase chain reaction (PCR) offers a specific and rapid diagnostic method for TSS . We describe a 7-year-old child with TSS resulting from staphylococcal pneumonia . S . aureus enterotoxins A and B were detected in the sputum of this patient by PCR. Presse Med, 2000 Dec 2, 29(37), 2023 - 7 {Resistance and new antibiotic strategies . New antistaphylococcal antibiotics}; Bergogne-Berezin E; INJECTABLE SPECTROGRAMIN: Combination regimens using quinupristin/dafopristin with either gentamicin or vancomycin have powerful bactericidal activities (even against quinupristin-resistant strains) against methicillin-resistant Staphylococcus aureus (MRSA) in a model of experimental endocarditis in the rabbit . In clinical trials, quinupristin/dalfopristin is becoming a therapeutic alternative to consider after failure of conventional antistaphylococcal treatments . NEW GENERATION CEPHALOSPORINS: These new cephalosporins, particularly C-3 pyridinium-thiomethyl-cephalosporins, new (3-dithiocarbamoyl) cephalosporins, and a series of new compounds with high affinity for MRSA PLP2a, are particularly active against MRSA and are unaffected by beta-lactamases . A NEW CARBAPENEM: This new antibiotic has a wide bactericidal effect against Gram-positive organisms and is active against MRSA as well as penicillin-resistant S . pneumoniae . NEW FLUOROQUINOLONE DERIVATIVES: In vitro, these new derivatives have been found to be active against MRSA, pneumococci non-sensitive to ciprofloxacin, and Bacteroides fragilis, Mycobacterium tuberculosis, Chlamydia pneumoniae. Presse Med, 2000 Dec 2, 29(37), 2018 - 21 {Resistance and new antibiotic strategies . The problem with staphylococcus}; Bergoge-Berezin E; MRSA outside the hospital: From a major problem almost exclusively encountered in hospitalized patients, methicillin-resistant Staphylococcus aureus (MRSA) infection has become a cause of skin, soft tissue, and even systemic infections outside the hospital . In order to prevent further spread, patients carrying MRSA (recently hospitalized patients, drug abusers, debilitated subjects, etc) must be identified and the mode of acquisition of MRSA infection . (cross transmission between community and non-hospital care centers) must be recognized . Small colony variants: SCV produce non-pigmented colonies that are 10 times smaller than the usual S . aureus colonies and have particular metabolic and genetic properties . They can cause persistent, recurrent and drug-resistant infections . MRSA in France: Data obtained in a case-control study confirms the selection pressure as well as the important colonization pressure exerted by antibiotics on gentamicin-sensitive strains. J Hum Lact, 2000 Nov, 16(4), 297 - 302 Lactation mastitis: bacterial cultivation of breast milk, symptoms, treatment, and outcome; Osterman KL et al.; The aim of this prospective study was to compare serum C-reactive protein (CRP) and leukocytes, hemoglobin, clinical signs, treatment, and outcome among 41 episodes of lactation mastitis grouped by the outcome of bacterial cultivation of breast milk . Group A included 25 cases with positive cultures only for bacteria normally present on skin . Group B included 16 cases in which cultures indicated the presence of potentially pathogenic bacteria . Serious complications were observed among women in group B, including protracted illness and weaning . No complications were observed in group A . Staphylococcus aureus was the most frequently isolated bacteria in group B . Mean serum leukocytes were significantly higher in group B than in group A . Although CRP levels in both groups were elevated, no significant difference was found between groups . Rest and frequent emptying of the breast were curative in group A . Further interventions were necessary for mothers in group B. J Viral Hepat, 2001 Jan, 8(1), 30 - 3 Activation-induced cell death in peripheral blood mononuclear cells (PBMCs) from patients with chronic hepatitis B may be related to abnormal production of interleukin 12 and 10; Ji W et al.; Peripheral blood mononuclear cells (PBMCs) from 70 patients with chronic hepatitis B and 32 normal healthy persons were isolated and cultured with or without Staphylococcus aureus enterotoxin B (SEB; 0.2 mg x l(-1)) and recombinant HBcAg (rHBcAg; 1.0 mg x l(-1)) for 48 h in vitro . After incubation, the cells were harvested by centrifugation and apoptosis of the PBMCs was studied by staining with fluorescent dyes YOPRO-1 and Hoechst 33342 . The levels of IL-12 and IL-10 in the serum and the supernatants of cultured PBMCs were assayed by ELISA . The levels of IL-12 heterodimer in the serum and the supernatants of PBMCs cultured with SEB or rHBcAg were lower in patients than controls . The levels of IL-10 in both the serum and supernatants were higher in patients than controls . In addition, the percentage of apoptotic cells in PBMCs from the infected patients was significantly greater than from normal persons in the presence or absence of SEB and rHBcAg . Patients seropositive for HBeAg had much greater percentage of apoptotic cells in the PBMCs cultured with rHBcAg than patients seronegative for HBeAg, reaching 24.08% . We speculate that activation-induced cell death of PBMCs in the patients with hepatitis B may be related to abnormal expression of IL-12 heterodimer and IL-10, which may lead to persistent infection in the patients. Clin Orthop, 2001 Jan, (382), 241 - 6 Collagen and fibronectin binding in experimental staphylococcal osteomyelitis; Johansson A et al.; A new mouse model of locally induced osteomyelitis was used to study the importance for pathogenicity of the specific binding ability of Staphylococcus aureus to collagen and fibronectin . This method appears to be convenient, reproducible, and suitable for large-scale experiments . In contrast to previous studies in experimental arthritis and endocarditis models, no difference in infection rates was found between the strains deficient in binding to collagen compared with the corresponding adherence-proficient strains . However, fibronectin binding ability in this model, in contrast to the endocarditis model, is thought to enhance the microorganisms' capacity to establish an infection . Infections caused by the fibronectin-binding strain also are thought to be clinically more aggressive than those caused by the nonbinding strain . Specific adherence mechanisms are thought to be operative in the pathogenesis of biomaterial associated osteomyelitis, and an improved understanding of such mechanisms may have an important prophylactic and therapeutic impact. Neurosurg Rev, 2000 Dec, 23(4), 175 - 204; discussion 205 Spinal epidural abscess: a meta-analysis of 915 patients; Reihsaus E et al.; Spinal epidural abscess (SEA) was first described in the medical literature in 1761 and represents a severe, generally pyogenic infection of the epidural space requiring emergent neurosurgical intervention to avoid permanent neurologic deficits . Spinal epidural abscess comprises 0.2 to 2 cases per 10,000 hospital admissions . This review intends to offer detailed evaluation and a comprehensive meta-analysis of the international literature on SEA between 1954 and 1997, especially of patients who developed it following anesthetic procedures in the spinal canal . In this period, 915 cases of SEA were published . This review is the most comprehensive literature analysis on SEA to date . Most cases of SEA occur in patients aged 30 to 60 years, but the youngest patient was only 10 days old and the oldest was 87 . The ratio of men to women was 1:0.56 . The most common risk factor was diabetes mellitus, followed by trauma, intravenous drug abuse, and alcoholism . Epidural anesthesia or analgesia had been performed in 5.5% of the patients with SEA . Skin abscesses and furuncles were the most common source of infection . Of the patients, 71% had back pain as the initial symptom and 66% had fever . The second stage of radicular irritation is followed by the third stage, with beginning neurological deficit including muscle weakness and sphincter incontinence as well as sensory deficits . Paralysis (the fourth stage) affected only 34% of the patients . The average leukocyte count was 15,700/microl (range 1,500-42,000/microl), and the average erythrocyte sedimentation rate was 77 mm in the first hour (range 2-50 mm) . Spinal epidural abscess is primarily a bacterial infection, and the gram-positive Staphylococcus aureus is its most common causative agent . This is true also for patients who develop SEA following spinal anesthetics . Magnetic resonance imaging (MRI) displays the greatest diagnostic accuracy and is the method of first choice in the diagnostic process . Myelography, commonly used previously to diagnose SEA, is no longer recommended . Lumbar puncture to determine cerebrospinal fluid protein concentrations is not needed for diagnosis and entails the risk of spreading bacteria into the subarachnoid space with consequent meningitis; therefore, it should not be performed . The therapeutic method of choice is laminectomy combined with antibiotics . Conservative treatment alone is justifiable only for specific indications . Laminotomy is a therapeutic alternative for children . The mortality of SEA dropped from 34% in the period of 1954-1960 to 15% in 1991-1997 . At the beginning of the twentieth century, almost all patients with SEA died . Parallel to improvements in the mortality rate, today more patients experience complete recovery from SEA . The prognosis of patients who develop SEA following epidural anesthesia or analgesia is not better than that of patients with noniatrogenic SEA, and the mortality rate is also comparable . The essential problem of SEA lies in the necessity of early diagnosis, because only timely treatment is able to avoid or reduce permanent neurologic deficits . The problem with spinal epidural abscesses is not treatment, but early diagnosis - before massive neurological symptoms occur" (Strohecker and Grobovschek 1986). J Antimicrob Chemother, 2001 Jan, 47(1), 83 - 6 In vitro synergy between cefepime and vancomycin against methicillin-susceptible and -resistant Staphylococcus aureus and Staphylococcus epidermidis; Lozniewski A et al.; The in vitro activity of cefepime combined with vancomycin was assessed by the chequerboard method against 35 clinical isolates of methicillin-susceptible (MSSA, n = 8) or -resistant (MRSA, n = 10) Staphylococcus aureus and methicillin-susceptible (MSSE, n = 9) or -resistant (MRSE, n = 8) Staphylococcus epidermidis and S . aureus ATCC 25923 (MSSA) . The combination was synergic against 16 isolates and additive/indifferent against 20 . For 10 of the clinical isolates (two MSSA, three MRSA, two MSSE, three MRSE) and the reference strain, the interaction of cefepime and vancomycin was also determined by the time-kill method . Except for one MRSA isolate, synergic killing was demonstrated with clinically achievable concentrations of vancomycin (0.5-1 mg/L) and cefepime (methicillin-susceptible isolates: 0.5-1 mg/L; methicillin-resistant isolates: 2-64 mg/L).
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