Panminerva Med, 2002 Sep, 44(3), 179 - 84 Gram-positive bacterial resistance . A challenge for the next millennium; Bassetti M et al.; Penicillin G was first used in 1941 . Since then, the trend in bacterial infections has changed . New antibiotics have been developed and bacterial resistance has spread as a consequence . The spread of Gram positive resistant bacteria is related to an inappropriate use of antibiotics . Antibacterial agents are abused or overused in various fields: medicine itself, veterinary science and zootechnics . Now, at the beginning of the third millennium we have been forced to limit our therapeutic options in order to combat these insidious enemies . Selective antibiotic pressure on the microbial population, notably on enterococci and staphylococci, made these two pathogens recalcitrant to traditional chemotherapy . It is a matter of concern that today, vancomycin-resistant Enterococcus spp . (VRE) and vancomycin-intermediate and resistant Staphylococcus aureus (VISA and VRSA) are now being observed worldwide among emerging pathogens . Most pharmaceutical companies are today developing antimicrobial drugs that are active against Gram-positive bacteria . Quinupristin/dalfopristin and linezolid are the most promising drugs and are available only for serious infections; future agents being developed for multi-resistant Gram-positive infections include daptomycin and the glycyclines, although these are still in the development phase . Nevertheless, our group has had the opportunity to treat some serious infections with these drugs and the good results achieved are reported in this review. Arch Surg, 2002 Jul, 137(7), 789 - 93 Male sex predisposes the newborn surgical patient to parenteral nutrition-associated cholestasis and to sepsis; Albers MJ et al.; HYPOTHESIS: Sepsis is an epiphenomenon of parenteral nutrition-associated cholestasis (PNAC) and not a causative factor, and the incidence of sepsis is not affected by the presence or absence of PNAC . DESIGN: Observational cohort study . SETTING: Pediatric surgery department in a tertiary referral children's hospital . PATIENTS: Newborns receiving PN for at least 7 days following intestinal surgery . MAIN OUTCOME MEASURES: The criteria for PNAC were as follows: PN for at least 14 consecutive days, conjugated bilirubin level greater than 1.5 mg/dL (>26 micromol/L), conjugated bilirubin fraction greater than 50%, and absence of another identifiable cause of cholestasis . The identification of septic events was based on Centers for Disease Control and Prevention criteria . RESULTS: The patients (26 with PNAC and 72 without PNAC) were well comparable for underlying disease, gestational age, birth weight, and age at the start of PN . Time receiving PN and length of hospital stay were significantly (P<.001) longer in patients with PNAC . Parenteral nutrition-associated cholestasis was associated with male sex (P =.03; odds ratio, 2.8; 95% confidence interval, 1.1-7.1) . The overall sepsis incidence was low (9 per 1000 hospital days) . The sepsis incidence tended to be higher in patients with PNAC than in patients without PNAC (11.8 vs 7.1 per 1000 days; P =.08), but was significantly higher in male than in female patients (12.2 vs 5.6 per 1000 days; P =.01) . Most septic events were caused by coagulase-negative staphylococci . CONCLUSIONS: Sepsis is an epiphenomenon of PNAC rather than a causative factor . Moreover, male sex predisposes the newborn surgical patient to PNAC and to sepsis. J Clin Microbiol, 2002 Jul, 40(7), 2659 - 61 Rapid identification of Staphylococcus aureus and the mecA gene from BacT/ALERT blood culture bottles by using the LightCycler system; Shrestha NK et al.; One hundred BacT/ALERT blood culture bottles growing gram-positive cocci in clusters were cultured and studied by LightCycler PCR for the sa442 and mecA genes . PCR was 100% sensitive and specific for detecting Staphylococcus aureus and methicillin resistance in S . aureus but was less accurate for methicillin resistance in coagulase-negative staphylococci. J Clin Microbiol, 2002 Jul, 40(7), 2520 - 5 Nosocomial spread of a Staphylococcus capitis strain with heteroresistance to vancomycin in a neonatal intensive care unit; Van Der Zwet WC et al.; A premature infant in a neonatal intensive care unit (NICU) developed a bloodstream infection caused by coagulase-negative staphylococci (CoNS) sensitive to vancomycin . The infection persisted for 3 weeks, despite therapy with vancomycin and replacement of all intravenous catheters . The neonate died due to necrotizing enterocolitis which developed during the ongoing sepsis . We screened this strain and 216 other strains of CoNS from cultures of blood obtained from neonates between 1997 and 2000 for heteroresistance to vancomycin . Forty-eight isolates, including the strain that caused ongoing sepsis, proved heteroresistant . All isolates were identified as Staphylococcus capitis and were identical, just as their resistant stable subcolonies were, when they were genetically fingerprinted by amplified-fragment length polymorphism analysis . The heteroresistant phenotype of this endemic strain was confirmed by population analysis . We conclude that heteroresistance to vancomycin occurs in S . capitis and might be the cause of therapeutic failures in NICUs . Moreover, heteroresistant strains can become endemic in such units. J Clin Microbiol, 2002 Jul, 40(7), 2480 - 2 Performance of CHROMagar selective medium and oxacillin resistance screening agar base for identifying Staphylococcus aureus and detecting methicillin resistance; Kluytmans J et al.; Two new selective media, oxacillin resistance screening agar base (ORSAB) and CHROMagar Staph aureus (CSA), were evaluated for identification of Staphylococcus aureus and for screening of methicillin resistance by addition of antimicrobial agents to these media . A well-defined collection consisting of 1,140 staphylococci was used . A total of 624 were S . aureus, of which 358 were methicillin susceptible and 266 were methicillin resistant, and 516 were coagulase-negative staphylococci . The methicillin-resistant S . aureus (MRSA) strains were selected based on the results of phage typing; 247 different types were included in the analysis . For identification of S . aureus, both media performed better after 24 h than after 48 h . The sensitivities at 24 h were comparable (CSA, 98.6%; ORSAB, 97.1%), but the specificity of CSA was significantly higher (CSA, 97.1%; ORSAB, 92.1%) . For screening of methicillin resistance, antibiotic supplements were added to both media . The sensitivity was lower after 24 h (CSA, 58.6%; ORSAB, 84.2%) and increased significantly after 48 h (CSA, 77.5%; ORSAB, 91.4%) . At both time intervals ORSAB was significantly more sensitive than CSA . However, the specificities of both media were high after 24 h (CSA, 99.1%; ORSAB, 98.3%) and decreased significantly after 48 h of incubation (CSA, 94.7%; ORSAB, 95.5%) . In conclusion, for identification of S . aureus, CSA is more accurate than ORSAB because of a significantly higher specificity . For screening of MRSA, ORSAB performs better than CSA, but the usefulness in clinical practice is limited because a significant number of strains are not detected. J Clin Microbiol, 2002 Jul, 40(7), 2392 - 7 Detection of methicillin-resistant Staphylococcus aureus and simultaneous confirmation by automated nucleic acid extraction and real-time PCR; Grisold AJ et al.; A molecular assay for the simultaneous detection of a Staphylococcus aureus-specific gene and the mecA gene, responsible for the resistance to methicillin in staphylococci, was evaluated . The assay included an automated DNA extraction protocol conducted with a MagNA Pure instrument and real-time PCR conducted with a LightCycler instrument . The performance and robustness of the assay were evaluated for a suspension of methicillin-resistant S . aureus (MRSA) strain with a turbidity equivalent to a McFarland standard of 0.5, which was found to be the ideal working concentration . The specificity of the new molecular assay was tested with a panel of 30 gram-negative and gram-positive bacterial strains other than MRSA . No cross-reactivity was observed . In a clinical study, 109 isolates of MRSA were investigated . All clinical MRSA isolates gave positive results for the S . aureus-specific genomic target, and all but one were positive for the mecA gene . In conclusion, the new molecular assay was found to be quick, robust, and laborsaving, and it proved to be suitable for a routine molecular diagnostic laboratory. Diagn Microbiol Infect Dis, 2002 Jun, 43(2), 135 - 40 Validation of Vitek version 7.01 software for testing staphylococci against vancomycin; Raney PM et al.; We tested 143 isolates of staphylococci with vancomycin by the National Committee for Clinical Laboratory Standards broth microdilution (BMD) reference method and compared the results to those generated using the Vitek automated system (GPS-105 and GPS-107 cards and version 7.01 software) . For ten isolates, the vancomycin MICs by BMD were 8 microg/ml . By Vitek, the vancomycin MICs ranged from 2 to 16 microg/ml . Vancomycin MICs of > or =32 microg/ml were reported for two additional isolates by Vitek; however, the MICs decreased to < or =0.5 microg/ml on retesting . By BMD, the vancomycin MICs for both isolates were 1 microg/ml . While the modal vancomycin MIC results by BMD for S . aureus and coagulase-negative staphylococci (CoNS) were both 1 microg/ml, Vitek results showed a mode of < or =0.5 microg/ml for S . aureus, and a mode of 2 microg/ml for CoNS . Vitek did not report vancomycin MICs of 1 or 4 microg/ml for any of the isolates tested . While the sensitivity of detecting staphylococci with reduced susceptibility to vancomycin appears to be improved with Vitek version 7.01 software, when compared to earlier software versions, laboratories may notice an overall shift in MIC data toward higher vancomycin MICs, although for the most part, this does not affect the categorical interpretations of the results. J Nat Prod, 2002 Jun, 65(6), 872 - 5 New antimicrobial cycloartane triterpenes from Acalypha communis; Gutierrez-Lugo MT et al.; Three new cycloartane-type triterpenes, 16 alpha-hydroxymollic (1), 15 alpha-hydroxymollic (2), and 7 beta,16 beta-dihydroxy-1,23-dideoxyjessic acids (3), were isolated from the aerial parts of Acalypha communis . The structures of the novel triterpenes were determined by spectroscopic methods as well as chemical derivatization . These compounds were tested for their antimicrobial activity against Gram-positive and -negative bacteria . Compounds 1-3 exhibited moderate antimicrobial activity (MIC 8, 32, 8 microg/mL, respectively) against vancomycin-resistant enterococci . In addition, compound 1 was found to be active against methicillin-resistant staphylococci . In contrast, compounds 1-3 were poorly active against Gram-negative bacteria . Compound 3 was tested in an in vivo model; it did not provide protection to mice infected with Staphylococcus aureus. Antibiot Khimioter, 2002, 47(2), 11 - 5 {Effect of peptide antibacterial factor and changes in Staphylococcus susceptibility to antibiotics}; Korobov VP et al.; The susceptibility to antibiotics of the Staphylococcus epidermidis cells in the presence of low-molecular weight peptide factor was investigated . The factor was isolated from the Styaphylococcus warneri culture media . The factor enhanced the S . epidermidis susceptibility to ampicillin, chloramphenicol, lincomycin, rifampicin, tetracycline, cefalexine, erythromycin and fusidine . The same effect was demonstrated for the cells of S . epidermidis resistant to cadmium ions along with resistance to several antibiotics . However the cells reaction in this case was expressed to a lower extent . It was suggested that revealed alterations in staphylococci susceptibility to antibiotics was due to the peptide factor effect on cytoplasmic membrane permeability that resulted in nonspecific increase in accumulation of various compounds with low-molecular weight. Clin Infect Dis, 2002 Jul 15, 35(2), e14 - 9 Epub 2002 Jun 18. Nosocomial bloodstream infections in Finnish hospitals during 1999-2000; Lyytikainen O et al.; Prospective laboratory-based surveillance in 4 Finnish hospitals during 1999-2000 identified 1477 cases of nosocomial bloodstream infection (BSI), with an overall rate of 0.8 BSIs per 1000 patient-days . Of BSI cases, 33% were in patients with a hematological malignancy and 15% were in patients with a solid malignancy; 26% were in patients who had undergone surgery preceding infection . Twenty-six percent of BSIs were related to intensive care, and 61% occurred in patients with a central venous catheter . Sixty-five percent of the 1621 causative organisms were gram positive, 31% were gram negative, and 4% were fungi . The most common pathogens were coagulase-negative staphylococci (31%), Escherichia coli (11%), Staphylococcus aureus (11%), and enterococci (6%) . Methicillin resistance was detected in 1% of S . aureus isolates and vancomycin resistance in 1% of enterococci . The 7-day case-fatality ratio was 9% and was highest for infections caused by Candida (21%) and enterococci (18%) . The overall rate of nosocomial BSIs was similar to rates in England and the United States, but S . aureus, enterococci, and fungi were less common in our study, and the prevalence of antibiotic resistance was lower. J Dairy Sci, 2002 May, 85(5), 1133 - 40 Influence of Staphylococcus aureus strain-type on mammary quarter milk somatic cell count and N-acetyl-beta-D-glucosaminidase activity in cattle from eight dairies; Middleton JR et al.; The hypothesis tested was that there are differences in pathogenicity between strains of Staphylococcus aureus that cause bovine mastitis . Mammary quarter milk somatic cell count (SCC) and N-acetyl-beta-D-glucosaminidase (NAGase) activity were used as indicators of the pathogenicity of different strains of S . aureus that infect the bovine udder . Eight commercial dairy herds with a history of S . aureus in bulk tank milk cultures were studied . Initially, composite foremilk samples were collected from all lactating cattle in each herd and cultured for staphylococci . Subsequently, all cows with a coagulase-positive staphylococcal intramammary infection (IMI) at the initial sampling that were still present in the herd of origin had individual mammary quarter foremilk samples collected . Coagulase-positive staphylococcal isolates were confirmed as S . aureus using a commercial biotyping system . Staphylococcus aureus isolates were strain-typed using pulsed-field gel electrophoresis . Mammary quarter milk SCC and N-acetyl-beta-D-glucosaminidase activity were determined for each cow . The difference in mean somatic cell count and mean NAGase activity for mammary quarters infected with the same strain of S . aureus and for uninfected quarters on the same cow was calculated . One-way analysis of variance was used to assess differences between strains within a herd . Overall, no significant differences were found between strains, suggesting that the degree of udder parenchymal injury induced by S . aureus IMI is in general significantly affected by factors other than strain type. Ann Fr Anesth Reanim, 2002 May, 21(5), 424 - 30 {Novel compounds active on staphylococci}; Fantin B; Research efforts to discover new compounds active against staphylococci are more than ever justified today . The incidence of methicillin-resistant staphylococci remains very high in hospitals, and the solution provided by glycopeptides is far from being satisfactory . These compounds exhibit mediocre pharmacokinetic and pharmacodynamic properties . Their ease and safety of use are poor . Finally, strains with diminished sensitivity to these antibiotics are beginning to appear . This article examines the opportunities offered by two new anti-staphylococcal agents: quinupristine-dalfopristine (Synercid) and linezolide (not marketed in France). Ann Fr Anesth Reanim, 2002 May, 21(5), 418 - 23 {The management of treatment failures for staphylococcal infections}; Wolff M; Therapeutic failure is a situation to be feared in severe infections due to staphylococci . In this article, we consider the difficulties faced is achieving a positive diagnosis in the context of such failures, and an approach to analysing their causes . The possible reasons for therapeutic failure must be considered systematically . The two principle causes are unquestionably the persistence in the patient of an infected, foreign body (intravenous catheter or prosthesis) and endovascular infections (thrombophlebitis or endocarditis) . The rules for the prescription of antibiotics are followed, target concentrations are achieved and drug failures are a rarity today . This observation may be called into question by the emergence of strains with diminished sensitivity to glycopeptides . These therapeutic failures remain exceptional cases today. Ann Fr Anesth Reanim, 2002 May, 21(5), 410 - 3 {Profile of patients facing therapeutic problems in nosocomial infections due to methicillin-resistant staphylococci--A survey of practices}; Sollet JP et al.; Methicillin-resistant staphylococcal infections (MRSI) are still common in an intensive care setting . Their management is mainly based on glycopeptides, combined with other antibiotics when this is possible, and also on treatment of the portal of entry (removal of foreign bodies, surgery...) . Implementation of this antibiotic therapy may meet with difficulties linked to the micro-organism (existence of strains with diminished sensitivity to glycopeptides), to the toxicity of glycopeptides or to the unfavourable course of the infection . A survey of practices was performed on a representative sample of 240 intensive care units in France . Glycopeptides, and particularly vancomycin, were the most frequently employed and prescribed in combination with other compounds . Therapeutic problems were considered as occasional, their incidence ranging from 0 to 50% . The problems reported were mainly linked to adverse effects: most frequently renal toxicity and, to a lesser extent, immunological and allergic complications . Diminished sensitivity to glycopeptides was only reported by a third of physicians, and this sporadically . Such a survey of practices is an essential preliminary to an epidemiological study of the incidence of MRSI and related therapeutic problems. Ann Fr Anesth Reanim, 2002 May, 21(5), 375 - 83 {Staphylococci resistant to antibiotic therapy}; Leclercq R; Increasing frequency in antibiotic resistance in Staphylococcus aureus is worrying . In this review, we briefly describe the mechanisms of resistance to antibiotics in staphylococci and we report the current incidence of antibiotic resistance in France . Methicillin-resistance is mostly confined to hospitals . However, spread of multiply resistant strains of staphylococci to the community is a putative threat . Strains with diminished susceptibility to glycopeptides are rare but possibly underestimated because of the in vitro difficulties to detect glycopeptide resistance . A continuous resistance survey is necessary . The problem of multiresistance is different for hospital and community staphylococci . The later strains still remain susceptible to many antibiotics, apart from some exceptions . By contrast, incidence of multiply resistance in hospitals remains high . After a recent decrease, incidence of oxacillin resistance seems to be stabilized . A new and recent feature is the great variety of resistance phenotypes which can be found in methicillin-resistant strains, in particular with gentamicin susceptibility. FEMS Microbiol Lett, 2002 Jun 18, 212(1), 35 - 9 A novel method for rapid production and purification of exfoliative toxin A of Staphylococcus aureus; Ladhani S et al.; The exfoliative toxins of Staphylococcus aureus are the causative agents of the scalded-skin syndrome . Previously described methods of toxin production and purification require large quantities of culture medium, take a long time and often produce low yields of toxin . A novel method of toxin production and purification using a dialysis sac to separate the culture medium from the staphylococci is described . This method produces up to 12 mg of crude toxin per ml of bacterial cell culture bathing the surface of the dialysis sac within 36 h and almost 10 mg of purified toxin per ml of cell culture within 3 days, in contrast to previous procedures that took over a week to produce 0.1-1.0 mg ml(-1) crude toxin and less than 0.01 mg ml(-1) purified toxin . This rapid method of toxin production should speed up future research into the pathogenesis of the staphylococcal scalded-skin syndrome. APMIS, 2002 Apr, 110(4), 332 - 9 Phenotypic and genotypic characterisation of blood isolates of coagulase-negative staphylococci in the newborn; Bjorkqvist M et al.; Coagulase-negative staphylococci (CNS) are the leading cause of late-onset sepsis in newborns (>72 h of age) . Our aim was to determine whether phenotypic and/or genotypic differences existed between blood isolates of CNS regarded as inducers of sepsis or as contaminants . Ninety-seven bloodisolates of CNS recovered from newborns at the neonatal intensive care unit, Orebro, Sweden in 1983-1997 were analysed . Twenty-nine of them (30%) were classified as sepsis isolates and 68 (70%) as contaminants . The most prevalent species was Staphylococcus epidermidis (n=59) . Staphylococcus haemolyticus (n=16) was most often isolated from newborns with the lowest gestational age and birth weight . Biochemical typing using the Phene Plate system (PhP) and genotyping using pulsed-field gel electrophoresis (PFGE) showed that the S . epidermidis isolates regarded as inducers of sepsis (n=16) were more homogeneous than isolates considered contaminants (n=37) . One main genotypic group, representing seven (44%) isolates, was identified among the sepsis isolates . Phenotypically the S . epidermidis sepsis isolates comprised three major clusters . In contrast, among the S . epidermidis contaminants, eight genotypic groups and two phenotypic clusters were identified . The dominating genotypic group among the sepsis isolates of S . epidermidis may represent strains with higher invasive capacity. Int J Food Microbiol, 2002 Jul 25, 77(1-2), 61 - 70 Staphylococcal enterotoxin genes of classical and new types detected by PCR in France; Rosec JP et al.; The PCR detection of the genes coding for the newly described staphylococcal enterotoxins (SE) G, H, I and J was carried out for 332 foodborne staphylococci, isolated from a variety of foods in France . The frequency of the Staphylococcus aureus strains harboring these genes was found to be very high (57%) and greater than that of the strains harboring "classical" SE genes as previously established . If one takes into account the newly described SE genes, in addition to the classical SE genes, the percentage of foodborne enterotoxigenic S . aureus strains doubles . The S . aureus biovars that were rarely or never enterotoxigenic (i.e., the poultry and bovine biovars) frequently become more potentially toxigenic, if taking into account the seg . seh, sei and sej genes . Furthermore, this work indicated a systematic association between the seg and sei genes and their high incidence among the S . aureus strains, which suggests that these two SE's could be an important phylogenetic link among the staphylococcal enterotoxins. Vet Dermatol, 2002 Jun, 13(3), 123 - 30 Colonization of neonatal puppies by Staphylococcus intermedius; Saijonmaa-Koulumies LE et al.; Cutaneous and mucosal carriage of Staphylococcus intermedius was determined in five Cavalier King Charles spaniel bitches, living in normal domestic environment, and their 18 puppies during the immediate prepartum period and after whelping, over a total of 8 weeks, using quantitative swab techniques . Within 8 h of birth S . intermedius was isolated from each puppy . The nose and mouth were the most frequently colonized mucosal sites (83 and 78%, respectively) . At that time S . intermedius was isolated from the abdominal skin in 78% of the puppies . One day postpartum the mucosal populations increased significantly (P < 0.001) but fell 6 (P < 0.01) and 20 (P < 0.001) days later . Populations from abdominal skin reduced during the first and fourth weeks postpartum (P < 0.05 for both) . This study indicates that in the domestic environment S . intermedius establishes itself soon after birth . The level of colonization by S . intermedius in the bitches seems to influence the colonization by pathogenic staphylococci in puppies. Int Orthop, 2002, 26(3), 179 - 84 Epub 2002 Apr 16. Infection after prosthetic reconstruction in limb salvage surgery; Lee SH et al.; Among 145 patients that underwent prosthetic reconstruction between 1985 and 1998, infection developed in 18 . Osteosarcoma was the most common primary tumour, and staphylococci were the major pathogenic organisms . Infections were controlled in five patients with debridement only and in two patients with a two-stage revision . Arthrodeses were performed in three and amputations in two . In six patients, infection was suppressed with antibiotics but not cured . Pulmonary metastasis developed in four of ten patients in whom chemotherapy was delayed or interrupted . Infection controls were poor in cases of delayed debridement over 2 weeks, in cementless fixation of implant, and soft tissue defects ( P<0.01) . Infection after prosthetic reconstruction is not only difficult to treat but also influences the prognosis reverse. Br J Dermatol, 2002 Jun, 146(6), 943 - 51 Invasion of human keratinocytes by Staphylococcus aureus and intracellular bacterial persistence represent haemolysin-independent virulence mechanisms that are followed by features of necrotic and apoptotic keratinocyte cell death; Mempel M et al.; BACKGROUND: Colonization of human skin by Staphylococcus aureus is a characteristic feature of several inflammatory skin diseases, which is often followed by tissue invasion and severe cell damage . A crucial role has been attributed to staphylococcal haemolysins in the cytotoxicity to epidermal structures . OBJECTIVES: To investigate haemolysin-independent virulence to human keratinocytes . METHODS: The stable alpha-haemolysin, beta-haemolysin double-negative S . aureus mutant DU 5720 was compared with the fully virulent parent strain 8325-4 and with its isogenic fibronectin-binding protein A/B-negative variant DU 5883 in an invasion model . RESULTS: This assay showed dose-dependent internalization of all the strains investigated by human HaCaT keratinocytes, with reduced internalization of DU 5883 . Transmission electron microscopy revealed adhesion of staphylococci to cellular pilus-like extrusions, followed by the embedding of the bacteria in cellular grooves . Following attachment to the keratinocytes the staphylococci were engulfed into vesicles within the cytoplasm where some bacteria persisted for 24-48 h . Addition of cytochalasin D strongly reduced the bacterial uptake, suggesting an active keratinocyte process . Bacterial invasion was followed by severe keratinocyte cell damage showing the morphological changes of cytotoxic and, to a lesser extent, apoptotic cell death as determined by the trypan blue exclusion test and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling assay . The highest levels of lethal cytotoxicity were observed in haemolysin-producing strains, whereas the induction of apoptosis seemed to depend on internalization . CONCLUSIONS: Staphylococcal invasion of human keratinocytes represents a potent staphylococcal virulence factor, which, independently of alpha- and beta-haemolysins, leads to necrotic and apoptotic cell damage. J Pharm Biomed Anal, 2002 Jun 20, 29(1-2), 173 - 81 Identification by LC/MS(n) of degradates of a novel carbapenem antibiotic in an aqueous matrix; Zhao ZZ et al.; Increased drug resistance in Staphylcocci and Enterococci to currently available antibiotics has significantly limited therapeutic options . Recently, a novel carbapenem antibiotic (Compound A) with a releasable side chain adjacent to the carbapenem was investigated to combat methicillin- and vancomycin-resistant Staphylococci and vancomycin-resistant Enterococci . The major advantage of Compound A over existing antibiotics can be attributed to the fact that cleavage of the side chain upon beta-lactam ring opening retained anti-bacterial activity while expelling the immunodominant epitope of the presumed beta-lactam hapten . In this work, LC/MS methods were developed to identify degradates of Compound A in an aqueous matrix utilized in assessing product safety and supporting analytical method and formulation development . A total of eight significant degradates were observed in this Compound A sample by LC/MS(n) and other techniques . Detailed structural analysis of degradates based upon LC/MS(n) data and other supporting results will be described in this work . Proposed molecular structures were confirmed by synthesis and use of authentic standards for several degradates . Degradates 1 and 4 were identified as degradates formed through the reversal of Michael reaction from Degradate 3 that is apparently formed by hydrolysis . Degradates 2 and 8 were found to be Hofmann elimination degradates . Degradates 5 and 6 are believed to be formed through dimerization of two parent molecules followed by the reversal of Michael reaction . Finally, Degradate 7 is attributed to a displacement reaction . Potential degradation pathways based upon these preliminary studies will also be discussed. FEBS Lett, 2002 Apr 24, 517(1-3), 72 - 8 Enhanced activation of bound plasminogen on Staphylococcus aureus by staphylokinase; Molkanen T et al.; Activation of plasminogen (plg) to plasmin by the staphylococcal activator, staphylokinase (SAK), is effectively regulated by the circulating inhibitor, alpha2-antiplasmin (alpha2AP) . Here it is demonstrated that intact Staphylococcus aureus cells and solubilized staphylococcal cell wall proteins not only protected SAK-promoted plg activation against the inhibitory effect of alpha2AP but also enhanced the activation . The findings suggest that the surface-associated plg activation by SAK may have an important physiological function in helping staphylococci in tissue dissemination . Amino acid sequencing of tryptic peptides originating from the 59-, 56- and 43-kDa proteins, isolated as putative plg-binding proteins, identified them as staphylococcal inosine 5'-monophosphate dehydrogenase, alpha-enolase, and ribonucleotide reductase subunit 2, respectively. Eur J Cardiothorac Surg, 2002 May, 21(5), 825 - 30 Postoperative mediastinitis in cardiac surgery - microbiology and pathogenesis; Gardlund B et al.; OBJECTIVE: During 1992-2000, postoperative mediastinitis developed after 126 (1.32%) of 9557 consecutive cardiac surgery procedures . The study was done to describe the variation in clinical characteristics and microbiological etiology in mediastinitis . METHODS: The records of 126 cases of postoperative mediastinitis were reviewed . RESULTS: The median time from operation to the development of mediastinitis was 7 days . Sternal dehiscence was seen in 86 patients (68%) . Coagulase negative staphylococci (CNS) were isolated in 46% of the cases with a verified microbiological etiology, Staphylococcus aureus in 26% and gram-negative bacteria in 18% . CNS were more frequently isolated in patients with sternal dehiscence (44/80, 55%) than in patients with stable sternum (10/38, 26%) (P=0.003) . However, S . aureus was more frequent in patients with stable sternum (18/38, 47%) than in patients with sternal dehiscence (13/80, 16%) (P<0.001) . High body mass index was associated with coagulase negative staphylococci (P<0.001) and with sternal dehiscence (P=0.008) . Chronic obstructive pulmonary disease was also associated with sternal dehiscence (P<0.001) and with coagulase negative staphylococci (P=0.04) . Patients who had been reoperated before onset of mediastinitis tended to have an increased risk for a gram-negative etiology (32 vs . 15% in patients not reoperated, P=0.06) . The overall 90-day all cause mortality in patients with mediastinitis was 19% . High age, need for reoperation before mediastinitis, and a long primary operation time was associated with increased mortality (P=0.02, P=0.007 and P=0.001, respectively) . No specific bacterial etiology was associated with increased mortality nor was the presence of bacteriemia . CONCLUSIONS: Three different types of postoperative mediastinitis can be distinguished: (1) mediastinitis associated with obesity, chronic obstructive pulmonary disease, and sternal dehiscence, typically caused by coagulase negative staphylococci; (2) mediastinitis following peroperative contamination of the mediastinal space, often caused by S . aureus, and (3) mediastinitis mainly caused by spread from concomitant infections in other sites during the postoperative period, often caused by gram negative rods . The proposed classification of mediastinitis into three groups with different pathogenic mechanisms may be useful in understanding which prophylactic counter measures have the potentials to be effective in a given situation. Clin Infect Dis, 2002 Jul 1, 35(1), 1 - 10 Epub 2002 Jun 04. Nosocomial spontaneous bacterial peritonitis and bacteremia in cirrhotic patients: impact of isolate type on prognosis and characteristics of infection; Campillo B et al.; The characteristics of and prognosis for nosocomial spontaneous bacterial peritonitis (SBP) and bacteremia were examined in a prospective study that included data from 194 consecutive episodes of SBP and 119 episodes of bacteremia, 93.3% of which were nosocomial, in 200 hospitalized cirrhotic patients . Gram-positive pathogens were predominant (70% of the total) among isolates from nosocomial infections; the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) was 24.8% . Nosocomial and staphylococcal infections were associated with a higher mortality rate than were community-acquired infections (P=.0255) and nonstaphylococcal infections (P<.001), respectively . In comparison with non-MRSA infections, MRSA infections were more likely to recur and occurred in a greater number of sites other than ascitic fluid and blood (P=.0004) . Older age (P=.0048), higher Child-Pugh score (P=.0011), and infection with staphylococci (P=.0031) were independently associated with a higher mortality rate . The emergence of MRSA is important because of the recurrence and poor outcome associated with infection with such organisms. Salud Publica Mex, 2002 Mar-Apr, 44(2), 108 - 12 Epidemiology of drug resistance: the case of Staphylococcus aureus and coagulase-negative staphylococci infections; Calderon-Jaimes E et al.; OBJECTIVE: To study the activity of several antibiotics against Staphylococcus spp . MATERIAL AND METHODS: The study included 1209 strains of Staphylococcus spp . from two institutions; Instituto Nacional de Pediatria (National Institute of Pediatrics) and Hospital Infantil de Mexico Federico Gomez (Mexico City Children's Hospital) . Minimum Inhibitory Concentrations of all antibiotics were determined by the agar macrodilution technique and standard methods from the National Committee for Clinical Laboratory Standards . RESULTS: Resistance of S . aureus was 14.2% and that of coagulase-negative staphylococci was 53.4% . The activity of different antibiotics is presented in detail . CONCLUSIONS: Surveillance of strains resistant to methicillin is necessary . The English version of this paper is available too at: http://www.insp.mx/salud/index.html. Diagn Microbiol Infect Dis, 2002 May, 43(1), 45 - 8 Agar diffusion, agar dilution, Etest, and agar screening test in the detection of methicillin resistance in staphylococci small star, filled; Rowe F et al.; Methicillin resistant Staphylococcus is an important worldwide problem . Resistance is verified in strains harboring the mecA gene and laboratory methods used to detect resistance are object of constant investigation . In the present study, 99 clinical isolates of staphylococci (41 S . aureus, 33 S . epidermidis, 12 S . saprophyticus and 13 members of other species) were submitted to different phenotypic methods and conditions . Detection of the mecA gene by PCR was used as the reference method and detected 14/41, 10/33, and 10/25 isolates of S . aureus, S . epidermidis and other species, respectively . Results showed that, for S . aureus and S . epidermidis, agar diffusion, agar dilution, and the E test incubated during 24h at 35 degrees C correctly discriminated mecA positive from mecA negative isolates . For other species, all methods and conditions presented low specificity (ranging from 20% to 66.7%) and, particularly S . saprophyticus, may need molecular methods to correctly assess methicillin resistance. Clin Microbiol Infect, 2002 May, 8(5), 310 - 5 The role of glycopeptides in the treatment of intravascular catheter-related infections; Graninger W et al.; There is increasing concern over multiresistant staphylococcci in catheter-associated infections . Local infections due to coagulase-negative staphylococci are usually resolved by removal of the intravascular catheter . However, if the device should remain for a certain period of time, e.g . to complete a course of chemotherapy, the antibiotic lock technique with a glycopeptide should be considered . In case of septic embolism to the lung caused by a multiresistant Staphylococccus aureus or Enterococcus faecium, systemic therapy with glycopeptides, streptogramins or linezolid must be employed. Clin Microbiol Infect, 2002 May, 8(5), 265 - 74 Catheter-related infections: diagnosis and intravascular treatment; Bouza E et al.; The diagnosis of catheter-related infections relies on the presence of clinical manifestations of infection and the evidence of colonization of the catheter tip by bacteria, mycobacteria, or fungi . The reference method to confirm the latter requires the withdrawal of the catheter for culturing, which frequently turns out to be inconvenient, unnecessary and costly . New methods try to avoid these drawbacks and to assess the presence of tip colonization without withdrawal . Comparative quantitative blood cultures with a marked increase (> or = 5) in colony counts between blood obtained from the catheter lumen and from a peripheral vein simultaneously is one of those methods . It has a high sensitivity (>80%) and specificity (94-100%) but it is cumbersome and requires both an easy backflow of blood in the catheter and the existence of bacteremia . Cytocentrifugation and acridine orange staining of blood withdrawn from an infected catheter lumen has a sensitivity and a specificity of over 90% for the diagnosis of tip colonization . 'Superficial cultures' comprise the semiquantitative culture of the hub, of the skin surrounding the catheter entrance and of the first subcutaneous portion (1 cm) of the catheter after swabbing . The sensitivity of this method is >90%, specificity is >80%, and positive and negative predictive values for catheters (considering together those with and without clinical data of infection) are 66 and 97%, respectively . Endoluminal brushing has proved to be an impractical and unreliable procedure, at least in our experience . New methods based on the speed of bacterial growth to detectable levels of micro-organisms in conventional blood cultures are a new and interesting way of assessing catheter-related infections . Moreover, as the use of antimicrobial-coated catheters becomes more prevalent, the existing definitions of catheter colonization and catheter-related infection may need to be modified, because such coatings may lead to false-negative culture results . Many catheter infections, diagnosed without catheter withdrawal, can be handled nowadays with the so-called 'antibiotic lock-in technique', which consists in 'locking' the infected catheter lumen with a solution containing antibiotics . A high proportion of infected catheters, mainly those with coagulase-negative staphylococci, can be maintained in place and sterilized with this technique, including catheters in patients with therapeutic failure after receiving conventional intravenous antibiotic therapy . New diagnostic and therapeutic techniques may avoid the unnecessary withdrawal of thousands of efficient, difficult to replace and expensive intravascular lines. J Chemother, 1991 Jan, 3 Suppl 1, 89 - 92 Teicoplanin and its influence on Staphylococcus aureus ultrastructure; Tullio V et al.; Teicoplanin, a glycopeptide antibiotic chemically related to the vancomycin-ristocetin group of antibiotics, has been shown to be active against gram-positive bacteria . Binding of teicoplanin by actively growing bacteria leads to morphological damage to the cell surface in staphylococci, as largely shown by transmission and scanning microscopy results. J Chemother, 1991 Jan, 3 Suppl 1, 7 - 14 Overview of septicemia; Venditti M et al.; Despite progress in antimicrobial therapy, septicemia remains a major problem of modern medicine . The clinical features and outcome may vary in different clinical settings and in a single setting during the years . As an example, gram-negative bacilli have been the prevalent cause of fulminant septicemia in granulocytopenic patients during the seventies . Nowdays, the use of indwelling central venous catheters and/or quinolone prophylaxis have favored the emergence of coagulase-negative staphylococci as a major cause of septicemia in these patients . As a consequence, the optimal management of febrile episodes in granulocytopenic patients should include not only a combination of a broad spectrum betalactam plus an aminoglycoside to prevent early death from gram-negative septicemia, but also antistaphylococcal antibiotics in cases not improving after 72 hours . The clinical spectrum of infective endocarditis continues to evolve . Infection of the right heart valves that was rare until a few decades ago, is now a frequent cause of staphylococcal septicemia in intravenous drug addicts . Along with prosthetic valve infection, new clinical syndromes of nosocomial endocarditis are emerging . Infections of permanent central venous catheters, ventriculoatrial shunts or pace-maker leads may in fact cause right-sided infective endocarditis . Septicemia will continue to challenge physicians in the future. J Chemother, 1991 Jan, 3 Suppl 1, 176 - 8 In-vitro activity of quinolones against different species of staphylococci of recent clinical isolation; Rabagliati AM et al.; We considered 256 strains of S . aureus (SA) and S . non-aureus (SnA) of recent clinical isolation . Their susceptibility (S) or resistance (R) to pefloxacin, ciprofloxacin, enoxacin, ofloxacin (which have been used in therapy for several years) and to timentin as well, has been tested, in relation with methicillin susceptibility (MS) and methicillin resistance (MR) . To date, the activity of quinolones against all staphylococci remains relatively high, notwithstanding the appearance of resistant strains, in particular among MR SnA . Of the four quinolones examined, ofloxacin showed to be the most active, i.e . effective against 100% of the staphylococci, either SA or SnA. J Chemother, 1991 Jan, 3 Suppl 1, 166 - 8 Selection of resistance to glycopeptide antibiotics in staphylococci; Biavasco F et al.; Clinical strains belonging to ten Staphylococcus species were investigated for their abilities to develop single-step resistance in vitro to vancomycin and teicoplanin . Surviving clones were only recovered from strains of three species, namely S . aureus, S . epidermidis, and S . haemolyticus . A similar ratio of grown to plated cells (approximately 1 x 10(-8)) was mostly obtained from strains of S . aureus and S . epidermidis . Higher ratios (1 x 10(-6) to 1 x 10(-7)) were obtained from strains of S . haemolyticus, especially when exposed to teicoplanin . When tested for susceptibility, many survivors exhibited vancomycin and teicoplanin minimum inhibitory concentrations (MICs) below the drug concentration used for in vitro selection, probably due to an inoculum effect in the plating procedure . However, MICs were particularly high in many clones of S . haemolyticus (up to 12.8 microg/ml for vancomycin, and > or = 102.4 microg/ml for teicoplanin). J Chemother, 1991 Jan, 3 Suppl 1, 144 - 8 The mechanism of staphylococci resistance to methicillin: a critical analysis of dominant opinions; Satta G et al.; Resistance to methicillin has been reported to be constantly associated with production of a novel penicillin-binding protein (PBP), indicated as PBP2a . This PBP is always present in strains that are homogeneously resistant to the drug, but is also constantly present in the strains heterogeneously resistant, where by far the vast majority of the cells are indeed sensitive to the antibiotic . This apparent lack of correlation between the presence of PBP2a and methicillin resistance has led to the idea, shared by most scientists, that the presence of PBP2a alone cannot explain resistance of staphylococci to methicillin . This confusion has implications also for clinical microbiology . In fact in in-vitro assays, not rarely, strains that are resistant to methicillin appear sensitive to other beta-lactams . However, clinical experience has demonstrated that when infections caused by methicillin-resistant staphylococci, which, in in vitro assays, become sensitive to other beta-lactams, are treated with the latter antibiotics, the incidence of failures is higher than that observed when the same antibiotics are used to treat infections caused by staphylococci sensitive to methicillin . Also in consideration of the fact that the mechanism of resistance to methicillin is not yet understood, many authors recommend to consider methicillin-resistant staphylococci as being in-vivo resistant also to the beta-lactams to which they may become sensitive in in-vitro assays . In the opinion of the authors, the mechanism of staphylococcal resistance to methicillin is reasonably explainable at least in its fundamental aspects . The present dominant opinions concerning methicillin-resistant staphylococci will be critically analyzed on the basis both of data presented in the literature and of data obtained in the authors' laboratory. J Chemother, 1991 Jan, 3 Suppl 1, 122 - 7 Effect of bacampicillin and ampicillin on the phagocytic, microbicidal functions of human macrophages; Cuffini AM et al.; During acute bacterial infections, antibiotics only assist the phagocytic system in eradicating the invading organism . In this regard, the effect of ampicillin and its prodrug bacampicillin on the phagocytosis and killing of Staphylococcus aureus by human macrophages was compared . Bacterial phagocytosis was enhanced after pre-incubation of staphylococci with bacampicillin . At a concentration of half the MIC bacampicillin showed a much more impressive killing capacity on macrophage-ingested live bacteria than did ampicillin, under all the experimental conditions tested. J Chemother, 1991 Jan, 3 Suppl 1, 119 - 21 Influence of teicoplanin on the chemiluminescence of phagocytosing human granulocytes; Aquilini D et al.; Teicoplanin is a new glycopeptide antibiotic that is structurally related to vancomycin . It is especially active against virtually all gram positive bacteria including methicillin-resistant staphylococci . The aim of this study was to evaluate the influence of teicoplanin on the phagocytosis of human neutrophils, using the chemiluminescence reaction method . We incubated three different concentrations (2, 5, 10 microg/ml) of teicoplanin for one and three hours with polymorphonuclear leukocytes of healthy human volunteers and then measured the chemiluminescence reaction during zymosan phagocytosis . The neutrophil function was not influenced by teicoplanin. J Clin Microbiol, 2002 Jun, 40(6), 2251 - 3 Detection of methicillin resistance in primary blood culture isolates of coagulase-negative staphylococci by PCR, slide agglutination, disk diffusion, and a commercial method; Hussain Z et al.; The methicillin resistance of 363 coagulase-negative staphylococci isolated from blood cultures was determined by a slide latex agglutination (LA) test for penicillin-binding protein 2a (PBP 2a), the presence of the mecA gene by PCR, disk diffusion, and Vitek . LA was performed on primary cultures, and PBP 2a expression was induced by placing an oxacillin disk in the primary inoculum . Compared to the PCR results, LA was the most sensitive and specific in the detection of methicillin resistance . Without induction, LA failed to detect 50% of mecA-positive strains grown on two different media. Breastfeed Rev, 2002 Mar, 10(1), 15 - 8 Breastfeeding and Staphylococcus aureus: three case reports; Amir L; This paper presents three case reports of breastfeeding women with Staphylococcus aureus (S . aureus) infections . The first case is a woman who developed recurrent staphylococcal skin infections, misdiagnosed as a fungal infection by her caregivers . The second case is a woman who experienced recurrent mastitis following a severe wound infection in her caesarean section scar; both she and her baby were carriers of S . aureus . The third case is a woman who experienced mastitis and a breast abscess, while her baby and other members of the family developed recurrent boils and skin infections with a methicillin-resistant S . aureus (MRSA) . A wide range of staphylococcal infections may occur in the postpartum period: mastitis, abscess, caesarean scar infection, boils and skin infection . Some cases of recurrent infections may be related to nasal carriage in mother or infant . Microscopy can be useful in differentiating bacterial infections from fungal infections and confirming nasal carriage . When mothers or infants are nasal carriers of Staphylococci health professionals may recommend nasal mupirocin (Bactroban) and bathing with antiseptic washes to reduce recurrent staphylococcal infections. Clin Exp Optom, 2002 May, 85(3), 172 - 5 Bacterial populations on high-Dk silicone hydrogel contact lenses: effect of length of wear in asymptomatic patients; Willcox MD et al.; BACKGROUND: Ocular infection and inflammation during extended wear with hydrogel lenses are often associated with contamination of the lenses with bacteria . This study examines the colonisation of a high-Dk silicone hydrogel contact lens worn on a 30-night extended wear basis (continuous wear) during asymptomatic lens wear . METHODS: Worn lenses were collected aseptically and placed in sterile vials . Microbial growth on various media was enumerated and the number of colony-forming units per whole lens was calculated . The proportions of samples contaminated with the bacteria and the extent of contamination were compared after one month, six months and two years of continuous wear with monthly replacement . The proportion of lenses that were sterile was calculated . RESULTS: The most frequently isolated and most numerous microbes that colonised the contact lenses during wear were the coagulase-negative staphylococci (in particular Staphylococcus epidermidis), followed by Propionibacterium sp . The frequency of isolation of the majority of bacterial isolates did not increase over time . Generally, lenses were sparsely colonised during asymptomatic wear . CONCLUSION: Up to two years continuous wear of silicone hydrogel lenses of the type used in this study does not appear to alter the types and number of bacteria that colonise the eye during wear, when lenses are replaced on a monthly schedule and the patients remain asymptomatic. J Food Prot, 2002 May, 65(5), 857 - 60 Incidence of enterotoxigenic staphylococci and their toxins in foods; Soriano JM et al.; Of 504 food samples collected from cafeterias, 19 (3.8%) yielded strains of enterotoxigenic staphylococci, and 10 (52.6%), 4 (21.1%), 3 (15.8%), and 2 (10.5%) of these strains produced enterotoxins C (SEC), D (SED), B (SEB), and A (SEA), respectively . Moreover, SEA, SEB, and SEC were isolated from three hamburger samples . Of 181 food samples collected from four restaurants before the implementation of the hazard analysis and critical control point (HACCP) system, 7 (3.9%) were found to contain enterotoxigenic strains, and SED, SEC, and SEA were produced by 4 (57.1%), 2 (28.6%), and 1 (14.3%) of these strains, respectively . One meatball sample with SEC was detected in a restaurant . After the implementation of the HACCP system in four restaurants, neither enterotoxigenic staphylococci nor enterotoxins were detected in 196 studied samples. Arch Dis Child, 2002 Jun, 86(6), 446 - 7 Clinical validation for oxacillin susceptibility testing of coagulase negative staphylococci; Cimolai N et al.; Oxacillin susceptibility of coagulase negative staphylococci as assessed by conventional methods was confirmed by a modified Etest method, extended to detect heteroresistance . Verification of susceptibility was followed by successful treatment for six consecutive children with deep seated infections . Physicians' trust in such a validated method will contribute to the appropriate use of antibiotics. Klin Lab Diagn, 2001 Mar, (3), 49 - 51 {Soybean bacteriological culture media and prospects for their use in clinical bacteriology}; Diachenko IuV et al.; Agar bacteriological nutrient media are suggested, based on soybean extract subjected to enzymatic hydrolysis and lyophilized native . The growth of test strains, number of grown colonies and their size were virtually the same as after inoculation of these strains in control nutrient media . Soybean salt medium with yolk suspension was tried with good results as elective medium for isolation of staphylococci from clinical material . Commercial manufacture of these media mainly from native soybean extract is proposed. Eur J Pediatr, 2002 May, 161(5), 270 - 4 Epub 2002 Mar 16. Whole blood platelet deposition on extracellular matrix under flow conditions in preterm neonatal sepsis; Finkelstein Y et al.; Platelet function in preterm infants with sepsis was evaluated by measuring their adhesion and aggregation properties using the Cone and Plate(let) Analyser . This may lead to earlier detection of bleeding tendency in septic infants . Platelet function was investigated in 54 preterm infants, of whom 32 had proven neonatal sepsis and 22 were healthy matched controls . Citrated whole blood was subjected to shear stress (1300 s(-1)) for 2 min on tissue culture plates precoated with subendothelial extracellular matrix (ECM) . The percentage of ECM surface covered with platelets and the average size of the ECM-bound platelet particles were determined with an image analyser . Assays for von Willebrand factor (vWF) antigen, ristocetin co-factor, and vWF collagen binding activity (CBA) were performed on samples from an additional 47 preterm infants: 38 healthy and 9 septic . Platelets of the preterm infants with sepsis displayed lower adhesion than those of the healthy controls . Mean surface coverage was 16.9+/-8.2% for the septic infants, 15.4+/-7.9% for the septic infants after exclusion of those with coagulase-negative staphylococci sepsis, and 20.8+/-9.6% for the healthy group ( P<0.05) . Platelet aggregation, vWF antigen, ristocetin co-factor, and CBA levels were similar between the septic and healthy groups . The most significant factor influencing surface coverage was the presence of sepsis . CONCLUSION: platelet adhesion to extracellular matrix is significantly lower in septic preterm infants than in healthy preterm infants . Intrinsic platelet properties, rather than the concentration or activity of plasma von Willebrand factor, may be responsible for this difference . Surface coverage obtained by the collagen binding activity test under flow conditions, which represents platelet adhesion, may be an earlier, more sensitive indicator of bleeding tendency in neonatal sepsis than decreased platelet count. J Hosp Infect, 2002 May, 51(1), 33 - 42 Molecular epidemiology of coagulase-negative staphylococcal bacteraemia in a newborn intensive care unit; Raimundo O et al.; We isolated 55 coagulase-negative staphylococci (CoNS) over two separate 12-month periods (26 in 1993 and 29 in 1996) from the blood of neonates in a neonatal intensive case unit (NICU) in Melbourne, Australia and compared them by pulse-field gel electrophoresis profile (PFGE), random amplification of polymorphic DNA (RAPD) and antibiogram . The most common species were Staphylococcus epidermidis, S . haemolyticus and S . warneri . The majority of such isolates were resistant to penicillin and to either or both of methicillin and gentamicin . During 1993, there was an increase in the number of CoNS bloodstream infections compared with previous years . S . epidermidis was the most common isolate, with 88% assessed as clinically relevant . Using the three typing systems, we identified one likely epidemic clone of S . epidermidis, the isolates of which were resistant to penicillin, gentamicin and erythromycin and possessed the mecA gene . There was complete correlation between the detection of mecA and the phenotypic expression of resistance when zone diameters in the disc diffusion assay were interpreted according to the latest NCCLS guidelines (1999) . Profiles of the remaining 1993 isolates were generally heterogeneous, suggesting independent acquisition with some evidence of cross-infection . The predominant bloodstream isolates in 1996 were heterogeneous multi-resistant strains of S . epidermidis, S . haemolyticus and S . warneri, about half of which were assessed as clinically relevant . These data support the view that CoNS are significant nosocomial pathogens in NICU and that resistant clones may be transmitted between babies . Molecular epidemiological tools are helpful for understanding transmission patterns and sources of infection, and are useful for measuring outcomes of intervention strategies implemented to reduce nosocomial CoNS sepsis . PFGE was found to be more discriminatory than RAPD, but the latter provides results in a more timely manner . Diagn Microbiol Infect Dis, 2002 Apr, 42(4), 291 - 4 Oxacillin susceptibility testing of coagulase-negative staphylococci using the disk diffusion method and the Vitek GPS-105 card( small star, filled); Woods W et al.; One hundred and ninety-three isolates of coagulase-negative staphylococci (CoNS) were tested against oxacillin by agar dilution, disk diffusion, and Vitek (GPS-105 card), and the presence of the mecA gene determined by multiplex PCR . The results obtained by all testing methods were in agreement for 190 isolates . Two mecA-negative isolates (S . lugdunensis and S . haemolyticus) had MICs of < or = 0.25 microg/ml by agar dilution and Vitek but were resistant by disk diffusion . One mecA-positive isolate was resistant by Vitek and disk diffusion but had an agar dilution MIC of < or = 0.25 microg/ml . For the species of CoNS tested, oxacillin susceptibility results obtained with the Vitek GPS-105 card and disk diffusion correlated well with results obtained by National Committee for Clinical Laboratory Standards agar dilution and with the presence of the mecA gene. Diagn Microbiol Infect Dis, 2002 Apr, 42(4), 283 - 90 Emerging elevated mupirocin resistance rates among staphylococcal isolates in the SENTRY Antimicrobial Surveillance Program (2000): correlations of results from disk diffusion, Etest and reference dilution methods; Deshpande LM et al.; Staphylococci cause one-third of all serious invasive infections in the SENTRY Antimicrobial Surveillance Program including bacteremias and lower respiratory tract infections . Staphylococci are also commensals of the skin and nasal passages; therefore, topical agents active against these organisms are valuable in preventing infections or transfer of the organisms between patients and/or health care workers . Mupirocin is a potent topical anti-staphylococcal compound, but its effectiveness has been compromised by emerging resistance . In early 2000, the SENTRY Program detected 302 mupirocin-resistant isolates (131 Staphylococcus aureus, and 171 coagulase-negative staphylococci {CoNS}) from the United States (19/25 medical centers), Canada (4/5), Latin America (3/9) and Europe (7/18) . One hundred sixty-eight mupirocin-resistant and 59 susceptible isolates were tested further by reference MIC, Etest (AB BIODISK, Solna, Sweden) and disk diffusion (5-microg) methods . Mupirocin resistance rates for blood stream infections varied by geographic area: for S . aureus from 1.9 to 5.6%, and for CoNS from 12.8 to 39.9% . Using elevated mupirocin MIC results, two resistant populations were noted: low-level resistance at 8-128 microg/mL and high-level resistance at > or = 1024 microg/mL . Acceptable correlation was observed between Etest and disk diffusion results (r = 0.84) without serious intermethod interpretive errors . High-level resistant isolates had heavy growth with no visible zone around the disk; low-level resistant isolates produced hazy zones of inhibition, and susceptible strains had clear zones of inhibition at > or = 17 mm . As mupirocin resistance can be plasmid-mediated, the prudent and appropriate use of this topical agent is important to minimize the ongoing development of resistance . Local surveillance for emerging mupirocin resistance appears warranted particularly in the United States and Canada, pragmatically using a disk diffusion test screening . Where more precise data are needed, the Etest is a very accurate method for distinguishing mupirocin low-level from high-level resistance patterns. Diagn Microbiol Infect Dis, 2002 Apr, 42(4), 279 - 82 Evaluation of an anti-PBP 2a slide latex agglutination test in coagulase-negative staphylococci isolated in Greek hospitals; Petinaki E et al.; Oxacillin resistance was examined in 258 coagulase-negative staphylococci from Greek hospitals . mecA DNA was detected in 168 isolates, which were also resistant to oxacillin by agar dilution and disk diffusion, according to the current NCCLS breakpoints . Both methods exhibited a relatively low specificity misclassifying 21 and 19 of the 90 mecA-negative isolates respectively as oxacillin resistant . In contrast, an anti-PBP 2a latex agglutination test, applied after induction by oxacillin, correctly classified 163 mecA-positive (sensitivity 97%) and 88 mecA-negative isolates (specificity 97.7%). EMBO J, 2002 May 15, 21(10), 2323 - 31 Substrate promiscuity of an aminoglycoside antibiotic resistance enzyme via target mimicry; Fong DH et al.; The misuse of antibiotics has selected for bacteria that have evolved mechanisms for evading the effects of these drugs . For aminoglycosides, a group of clinically important bactericidal antibiotics that target the A-site of the 16S ribosomal RNA, the most common mode of resistance is enzyme-catalyzed chemical modification of the drug . While aminoglycosides are structurally diverse, a single enzyme can confer resistance to many of these antibiotics . For example, the aminoglycoside kinase APH(3')-IIIa, produced by pathogenic Gram-positive bacteria such as enterococci and staphylococci, is capable of detoxifying at least 10 distinct aminoglycosides . Here we describe the crystal structures of APH(3')-IIIa in complex with ADP and kanamycin A or neomycin B . These structures reveal that the basis for this enzyme's substrate promiscuity is the presence of two alternative subsites in the antibiotic binding pocket . Furthermore, comparison between the A-site of the bacterial ribosome and APH(3')-IIIa shows that mimicry is the second major factor in dictating the substrate spectrum of APH(3')-IIIa . These results suggest a potential strategy for drug design aimed at circumventing antibiotic resistance. Chest, 2002 May, 121(5), 1618 - 23 Incidence and outcome of polymicrobial ventilator-associated pneumonia; Combes A et al.; STUDY OBJECTIVE: To determine the epidemiology and outcome of polymicrobial ventilator-associated pneumonia (VAP) . SETTING: Two ICUs (18 and 17 beds) in a university hospital . DESIGN AND PATIENTS: We undertook a 16-month study of 124 patients in whom a first episode of VAP had been diagnosed . Patients in whom there was a suspicion of clinical or radiologic VAP underwent bronchoscopy, and VAP was confirmed by the presence of at least two of the following criteria: > or = 2% of cells with intracellular bacteria found on direct examination of BAL fluid (BALF); protected-specimen brush sample culture with > or = 10(3) cfu/mL; or BALF culture with > or = 10(4) cfu/mL . RESULTS: Monomicrobial infections were diagnosed in 65 patients (52%), and polymicrobial infections were diagnosed in 59 patients (48%) . Two different bacteria were isolated in 42 patients (34%), three different bacteria were isolated in 10 patients (8%), and four different bacteria were isolated in 7 patients (6%) . Patients' clinical characteristics at ICU admission and on the day of bronchoscopy were similar, particularly the prior duration of mechanical ventilation (MV), the type of ICU admission, disease severity scores, and antibiotic therapy received before VAP was diagnosed . The percentages of nonfermenting, Gram-negative bacilli and methicillin-resistant staphylococci involved in monomicrobial and polymicrobial episodes were similar . Furthermore, no significant difference was detected in outcome parameters, specifically in the mortality rate at 30 days, the ICU mortality rate, the duration of MV, and the rate of infection relapse . CONCLUSION: In our study population, the epidemiology and outcomes of patients with monomicrobial and polymicrobial VAP did not differ significantly. Microb Drug Resist, 2002 Spring, 8(1), 55 - 9 Validation of rapid screening tests for the identification of methicillin resistance in staphylococci; van Leeuwen WB et al.; The Velogene Rapid MRSA Identification Assay (Alexon-Trend Inc., Ramsey, MN), a commercially available 90-min genotypic test using a chimeric probe for the cycling-mediated recognition of the mecA gene in staphylococci, was compared with the MRSA-Screen latex agglutination test, a 15-min phenotypic test (Denka Seiken Co., Tokyo, Japan) for the identification of the mecA gene product PBP 2a . The results of both techniques were compared with mecA gene PCR . A total of 210 stock-culture strains were tested, consisting of 108 methicillin-susceptible Staphylococcus aureus (MSSA) strains and 92 methicillin-resistant S . aureus (MRSA) strains . The performance of the assays was good, displaying sensitivities and specificities for Velogene and MRSA Screen of 96.7% and 100% and 96.7% and 100%, respectively . Both Velogene and MRSA Screen could not correctly identify three of the MRSA strains each . Repeat testing with a larger inoculum or exposure of the three distinct strains to methicillin, respectively, resolved these problems . All MSSA strains as well as the other genera were correctly addressed by both techniques . The 10 methicillin-resistant Staphylococcus epidermidis strains were detected by both techniques . Both the Velogene and the MRSA Screen assays accurately identified mecA-positive staphylococcal strains and can be successfully used for routine application in clinical microbiology laboratories. Curr Microbiol, 2002 Jun, 44(6), 385 - 90 Susceptibility of Brazilian staphylococcal strains to glycopeptides evaluated by different testing methods; Ferreira Nunes AP et al.; Reports of staphylococci with reduced susceptibility to glycopeptides are cause for concern . This study evaluated the susceptibility of 84 staphylococci clinical isolates to glycopeptides by the disk diffusion, agar dilution, E-test, and BHIA screening methods . Vancomycin agar dilution showed all strains presented minimum inhibitory concentration (MIC) ranging from 0.5 to 2 microg/ml, and the E-test showed similar results . Teicoplanin agar dilution test showed MICs ranging from < or = 0.5 to 2 microg/ml for Staphylococcus aureus and MICs ranging from <0.25 to 32 microg/ml for coagulase-negative Staphylococcus (CNS) . Ten CNS isolates presented MICs ranging from 8 to 32 microg/ml for agar dilution and/or E-test . All the staphylococci were susceptible to vancomycin by the disk diffusion test (DDT), but two CNS isolates presented intermediate resistance to teicoplanin by the DDT and MICs of susceptibility, with two other CNS strains, teicoplanin-susceptible by the DDT, presented MICs of intermediate resistance . On the vancomycin-containing agar, 20 CNS isolates were able to grow, but no S . aureus strain . All these isolates showed MICs to teicoplanin (4-32 microg/ml) higher than those isolates that did not grow on the agar screen plate . PFGE of chromosomal SmaI digests showed a wide diversity of these CNS strains, without any predominance of a single PFGE pattern. J Appl Microbiol, 2002, 92 Suppl, 158S - 62S Susceptibility of antibiotic-resistant cocci to biocides; Fraise AP; Biocide resistance has hitherto been a poorly studied subject, possibly due to the belief that such resistance was rare and clinically insignificant . Various recent findings, however, have underlined the importance of biocide resistance as a clinically relevant phenomenon . Outbreaks of biocide-resistant organisms in hospitals have been described and the genetic mechanism for resistance to quaternary ammonium compounds (QACs) in Staphylococcus aureus has now been elucidated . Mycobacteria resistant to commonly used endoscope disinfectants are now commonly reported and have caused numerous adverse clinical events . Cross-resistance between triclosan and antituberculous drugs has been demonstrated in other strains of mycobacteria . This is related to a common mechanism of action . The work presented here describes studies into the biocide resistance of antibiotic-resistant cocci and attempts to create biocide-resistant strains in vitro . Strains of staphylococci (including methicillin-resistant Staph . aureus (MRSA)) and enterococci (including vancomycin-resistant enterococci (VRE)) had their susceptibility to biocides assayed using broth macro dilution methods and resistant strains were selected by serial subculture on biocide-containing media . Mutants were created with relative ease; for instance, triclosan minimal bactericidal concentrations (MBCs) increased from 0.002 to 3.12 mg l(-1) . Some strains of MRSA which have intermediate resistance to glycopeptides were demonstrated to have decreased susceptibility to some biocides . Biocide resistance amongst enterococci was demonstrated although there was no clear correlation between biocide and antibiotic resistance . The exact mechanisms of resistance in these strains are still being studied but it is clear that biocide resistance is an important clinical phenomenon. J Vet Med Sci, 2002 Mar, 64(3), 245 - 50 Isolation and species distribution of staphylococci from animal and human skin; Nagase N et al.; From April 1999 to December 2000, a survey was made on the distribution of Staphylococcus species on the skin of 7 kinds of animals and humans . Staphylococci were isolated from 12 (100%) of 12 pigs, 17 (89.5%) of 19 horses, 30 (100%) of 30 cows, 73 (90.1%) of 81 chickens, 10 (40%) of 25 dogs, 23 (76.7%) of 30 laboratory mice, 20 (52.6%) of 38 pigeons, and 80 (88.9%) of 90 human beings . The predominant staphylococci isolated from a variety of animal species were novobiocin-resistant species, S . xylosus and S . sciuri regardless of the animal host species . The novobiocin-resistant species including S . xylosus and S . sciuri were only occasionally isolated from human skin . The predominant staphylococci found on human skin were novobiocin-sensitive species, S . epidermidis (63.8%), followed by S . warneri (28.8%) and S . hominis (13.8%) . The results suggest that the staphylococcal flora inhabiting animal skin are different from those of human skin in regard to the predominant species isolated . In this study, we used pulsed-field gel electrophoresis to examine the chromosomal polymorphisms of S . epidermidis isolated most frequently from human skin . Strains of S . epidermidis showed the greatest genomic diversity in their fragment patterns. J Immunol, 2002 May 15, 168(10), 5207 - 12 Critical role of signaling through IL-1 receptor for development of arthritis and sepsis during Staphylococcus aureus infection; Hultgren OH et al.; IL-1R-deficient mice (IL-1R(-/-)) and their wild-type controls (IL-1R(+/+)) were i.v . inoculated with 1 x 10(7) or 10(6) Staphylococcus aureus per mouse to mimic bacterial sepsis and septic arthritis . The disease outcome was severely worsened in the IL-1R(-/-) mice as compared with IL-1R(+/+) mice . Indeed, 3 days after inoculation of 10(7) S . aureus per mouse 84% of IL-1R(-/-) mice displayed clinical signs of septicemia as compared with none of the IL-1R(+/+) mice . On day 9 after inoculation with 10(6) S . aureus per mouse 75% of the IL-1R(-/-) mice were dead as compared with none of the IL-1R(+/+) mice . Also, the number of staphylococci in circulation was 25- to 30-fold increased in IL-1R(-/-) mice as compared with IL-1R(+/+) mice, the most probable reason for the outcome . The frequency and severity of septic arthritis were significantly increased in IL-1R(-/-) mice, as compared with IL-1R(+/+) mice, following i.v . inoculation of staphylococci . This was probably due to an increased accumulation of bacteria in the joints of IL-1R(-/-) mice as compared with their wild-type controls . Interestingly, while serum levels of IL-18 in IL-1R(-/-) mice were significantly lower than in IL-1R(+/+) mice 24 h after inoculation of S . aureus, both IL-18 and IL-1beta were significantly increased in IL-1R(-/-) vs IL-1R(+/+) mice 4 days after the bacterial inoculation . In conclusion, IL-1R signaling plays a crucial role in host protection during systemic S . aureus infection as seen by the fatal outcome of S . aureus sepsis and arthritis in IL-1R-deficient mice. J Hosp Infect, 2002 Jan, 50 Suppl A, S9 - 12 Surgical prophylaxis in practice; de Lalla F; The use of antibacterial prophylaxis of postoperative infections is firmly established within clean-contaminated procedures . For clean procedures, prophylaxis has traditionally been reserved for operations involving foreign-body implantation . However, evidence that postoperative infections from non-prosthetic clean procedures are highly under-reported suggests that prophylaxis is also advisable, at least for some non-prosthetic procedures, such as breast surgery and herniorrhaphy . Although cefazolin is recommended by current guidelines, cefuroxime and cefamandole have a broader antimicrobial spectrum and should be preferred in clean prosthetic surgery prophylaxis . In this type of surgery glycopeptides are not recommended for routine use but may have a role for major prosthetic implantation in units with a high incidence of methicillin-resistant staphylococci . In the case of clean-contaminated procedures, cefazolin is recommended for routine use, although colorectal procedures require an agent with improved anti-anaerobic activity . In addition, experience has shown that obstetric/gynaecological, gastroduodenal and biliary tract surgery and appendectomy all require broad-spectrum antibacterial prophylaxis . Suitable agents include cefoxitin, cefotetan, ureidopenicillins and beta-lactam/beta-lactamase inhibitor combinations . The traditional surgical classification scheme needs to be replaced with a classification that additionally accounts for patient-specific risk factors . The limitations of the current scheme may partly explain why current guidelines are so seldom followed in clinical practice. J Infect Dis, 2002 May 15, 185(10), 1417 - 24 Epub 2002 Apr 22. On the role of Staphylococcus aureus sortase and sortase-catalyzed surface protein anchoring in murine septic arthritis; Jonsson IM et al.; Anchoring of Staphylococcus aureus surface protein to the cell wall is catalyzed by sortase, a transpeptidase . The contribution of staphylococcal surface proteins to establishment of infection was examined using a murine septic arthritis model . Intravenous inoculation of mice with the sortase-deficient mutant S . aureus strain SMK3 did not result in weight loss or severe septic arthritis, in contrast to the parent strain, S . aureus Newman . Direct inoculation of the sortase mutant into joint cavities also failed to cause severe synovitis or erosive arthritis . Furthermore, intravenous inoculation with staphylococci resulted in the rapid clearing of the sortase mutant from the bloodstream . This phenomenon demonstrates the involvement of host neutrophils; when these cells were depleted, sortase mutant staphylococci caused severe systemic infection, although not septic arthritis . These results suggest that sortase mutant staphylococci are significantly less virulent than the parent strain, Newman: the sortase mutant has decreased ability to reach target organs and, once there, to induce an inflammatory response. Curr Opin Infect Dis, 2000 Dec, 13(6), 593 - 598 New Gram-positive agents in nosocomial infection; Patterson JE; Multidrug-resistant Gram-positive pathogens such as vancomycin-resistant enterococci, methicillin-resistant Staphylococcus aureus, and methicillin-resistant coagulase-negative staphylococci account for a significant number of nosocomial infections, and new options for therapy have been lacking . During the past year, two agents have been released that have activity against these organisms: quinupristin/dalfopristin and linezolid . Although neither agent is a panacea, these recently released agents offer new options for therapy. J Korean Med Sci, 2002 Apr, 17(2), 168 - 72 Early Screening of oxacillin-resistant Staphylococcus aureus and Staphylococcus epidermidis from blood culture; Jeong J et al.; The timely detection of blood-borne pathogens is one of the most important functions of the microbiology laboratory . Recently, methicillin-resistant staphylococci have become the most important pathogens seen by the laboratory . The purpose of this study was to evaluate Staphy agar, a novel screening medium, for the detection methicillin-resistant Staphylococcus aureus, S . epidermidis, or other coagulase-negative staphylococci (CNS) from positive blood cultures showing Gram-positive cocci in clusters . Eighty-six blood cultures that yielded Gram-positive cocci in clusters were included in this study . The organisms were finally identified by the Vitek system, and oxacillin resistance was confirmed by polymerase chain reaction (PCR)-based mecA gene detection . The identification and oxacillin resistance of all S . aureus strains showed complete agreement with the Vitek and PCR results . The presumptive detection of S . epidermidis and other CNS were consistent with the Vitek system in 94.7%, and the screening of oxacillin resistance was consistent with the result of PCR in 92.1% of 38 strains . The Staphy agar method is reliable and rapid for differentiating Gram-positive cocci in clusters in blood and for determining their methicillin resistance. Antimicrob Agents Chemother, 2002 May, 46(5), 1561 - 3 Comparative pharmacodynamics of three newer fluoroquinolones versus six strains of staphylococci in an in vitro model under aerobic and anaerobic conditions; Wright DH et al.; Six strains of staphylococci were exposed to levofloxacin, moxifloxacin, or trovafloxacin in an in vitro pharmacodynamic model under both aerobic and anaerobic conditions . Each agent demonstrated a rapid 3-log(10) kill versus susceptible isolates regardless of condition . Against clinical isolates with reduced susceptibility, regrowth occurred by 24 h and was frequently associated with further increases in MICs. Pol Merkuriusz Lek, 2002 Jan, 12(67), 73 - 6 {The safety of patients and staff in the operating room}; Pawinska A et al.; The risk of postoperative infection is determined by a number of factors including the underlying health of the patient, the type of surgery and the environment in which the procedure take place . The predominant infective organisms are Staphylococci . Coagulase-negative staphylococci are regarded as emerging pathogens, especially in clean surgery . The operating team is the major source of the contaminated bacteria . Bacteria dispersed from the skin is of greater importance than bacteria from the respiratory tract, so masks are not always necessary in the theatre during an operation except by those at the table . Masks that function as protective barriers are another emerging issue . Due to a greater awareness of HIV and other blood-borne viruses, masks are taking on a greater role in protection health care workers from potentially infectious blood and body fluids . Bloodborne infection pose a large risk to healthcare workers . The incidence of occupational HBV infection has decline as a result of vaccine-induced immunity . Future vaccines against HIV and HCV would be a boon to healthcare workers. Int Orthop, 2002, 26(1), 40 - 3 The outcome of perioperative wound infection after total hip and knee arthroplasty; Abudu A et al.; Forty-one consecutive patients with primary knee arthroplasty and 37 with primary hip arthroplasty, all with perioperative wound infections, were followed for 50 (12-130) months . Staphylococci (coagulase negative and positive) accounted for 74% of wound infections . Mixed organisms accounted for 10% . Prosthetic infections developed in eight patients and aseptic loosening in three patients . All the prosthetic infections developed within 6 months of the primary surgery . Organisms responsible for superficial infections were responsible for prosthetic infection in five patients; no organisms were isolated in the remaining three patients . The presence or absence of wound dehiscence, wound haematoma, and postoperative pyrexia did not predict the development of deep sepsis; however, the presence of wound discharge was a significant risk factor. Vasa, 2002 Feb, 31(1), 48 - 55 Surgical management of prosthetic vascular graft infection: comparative retrospective analysis of 30 consecutive cases; Gassel HJ et al.; BACKGROUND: Graft infection after vascular prosthetic reconstruction for the treatment of peripheral arterial occlusive disease (PAOD) is a rare but severe complication with poor outcome . The options for surgical treatment are not uniformly accepted and remain controversial . PATIENTS AND METHODS: We retrospectively analyzed the histories of 30 patients treated for prosthetic graft infection (Szilagyi grade III) in our hospital between 1994 and 1999 to determine which forms of treatment were best suited for which types of patient . In the majority of cases the initial treatment was lower-extremity bypass surgery . The most frequent location of infection was the groin (73%) . Staphylococci (13% of which were methicillin resistant) were the most common type of bacteria . The overall incidence of prosthetic infection was 2.3% . RESULTS: After confirmation of the infection by computed tomography (CT) or white blood cell scintigraphy, one of the following 5 forms of surgical treatment was performed: 1 . Removal of the infected prosthesis and its simultaneous replacement by an autologous vein bypass . 2 . Bypass removal and secondary replacement by an autologous vein . 3 . Extra-anatomical replacement . 4 . Graft removal and primary amputation . 5 . Local therapy with debridement and secondary wound healing . In some patients primary amputation after graft infection was necessary to prevent further deterioration with fulminant sepsis . The overall mortality was 17%, the amputation rate was 60% . CONCLUSIONS: The best results were achieved by early complete removal of the alloplastic material and one-step replacement by either an autologous vein or extra-anatomic bypass . This resulted in a limb salvage rate of 54% and 40% and mortality rates of 9% and 0%, respectively. Zh Mikrobiol Epidemiol Immunobiol, 2002 Jan-Feb, (1), 73 - 6 {Antilysozyme activity and antibiotic resistance of periulcerous zone microflora in patients with gastric and duodenal ulcer}; Chervinets VM; Antilysozyme activity (ALA) as well as antibiotic resistance were detected in 133 microbial cultures isolated from bioptic specimens of the mucous membrane of the ulcerous and periulcerous zones, taken from patients with gastric and duodenal ulcer . In 85.7-94.7% of cases Gram positive cocci and in 62.5% of cases Gram-positive bacilli showed no ALA . 50% of Gram negative bacteria cultures lacked ALA, while the remaining 50% exhibited this activity, on the average, 2.36 +/- 1.40 mkg/ml . Lysozyme activity was determined in 33.3% of the isolated staphylococci strains both with and without ALA . Staphylococci isolated from the gastric mucosa of healthy controls had no ALA in 33.3% of cases, and in 66.7% of cases ALA was equal to 2 mkg/ml . Gram positive coccal microflora showed, mainly, high sensitivity to antibiotics . In Gram negative bacteria antibiotic resistance was determined in 44.3 +/- 21.2% of the isolates . In Gram negative microorganisms correlation between ALA and antibiotic resistance was observed . From the periulcerous zone of patients with gastric and duodenal ulcer persistence associated Gram negative microorganisms were mainly isolated. J Bacteriol, 2002 May, 184(9), 2465 - 72 The superoxide dismutase gene sodM is unique to Staphylococcus aureus: absence of sodM in coagulase-negative staphylococci; Valderas MW et al.; Superoxide dismutase (SOD) profiles of clinical isolates of Staphylococcus aureus and coagulase-negative staphylococci (CoNS) were determined by using whole-cell lysates and activity gels . All S . aureus clinical isolates exhibited three closely migrating bands of activity as previously determined for laboratory strains of S . aureus: SodM, SodA, and a hybrid composed of SodM and SodA (M . W . Valderas and M . E . Hart, J . Bacteriol . 183:3399-3407, 2001) . In contrast, the CoNS produced only one SOD activity, which migrated similarly to SodA of S . aureus . Southern analysis of eight CoNS species identified only a single sod gene in each case . A full-length sod gene was cloned from Staphylococcus epidermidis and determined to be more similar to sodA than to sodM of S . aureus . Therefore, this gene was designated sodA . The deduced amino acid sequence of the S . epidermidis sodA was 92 and 76% identical to that of the SodA and SodM proteins of S . aureus, respectively . The S . epidermidis sodA gene expressed from a plasmid complemented a sodA mutation in S . aureus, and the protein formed a hybrid with SodM of S . aureus . Both hybrid SOD forms as well as the SodM and SodA proteins of S . aureus and the S . epidermidis SodA protein exist as dimers . These data indicate that sodM is found only in S . aureus and not in the CoNS, suggesting an important divergence in the evolution of this genus and a unique role for SodM in S . aureus. Mikrobiol Z, 2001 Nov-Dec, 63(6), 19 - 24 {Oxidative activity and accumulation of metabolites in gonococcal and staphylococcal plasmid-containing strains}; Krysenko AV et al.; A higher rate of glucose oxidation by antibiotic-resistant staphylococcus and gonococcus strains has been established; at the same time the sensitive strains are characterised by more intensive oxidation of pyruvate and acetate as compared with the resistant strains . High rate of lactate accumulation by antibiotic-resistant strains has been marked for the studied staphylococci strains . Antibiotic-resistant strains of gonococci are characterised by more intensive accumulation of acetate as the end product of glucose catabolism . No significant differences were revealed for the stable and sensitive strains of gonococci as to the rate of acetate involving in metabolic processes, but a steady tendency to the increase of acetate consumption intensity is observed. Crit Care Med, 2002 Apr, 30(4), 833 - 6 Unexpected high risk of contamination with staphylococci species attributable to standard preparation of syringes for continuous intravenous drug administration in a simulation model in intensive care units; van Grafhorst JP et al.; OBJECTIVE: To determine the risk of bacterial contamination of the infusate in a simulation model of syringes prepared for continuous intravenous drug administration by intensive care unit nurses . Widely accepted standard procedures in the intensive care unit were compared with syringes prepared by pharmaceutical technicians working under standard aseptic conditions according to national guidelines . DESIGN: Prospective study . SETTING: Intensive care units of four general teaching hospitals and two university hospitals . SUBJECTS: We examined 650 syringes prepared from 10-mL ampules and 100 syringes prepared from rubber-compound-capped 50-mL vials by intensive care unit nurses of six hospitals . Also, 100 syringes from 10-mL ampules and 100 syringes from rubber-compound-capped 50-mL vials were prepared by pharmaceutical technicians . INTERVENTIONS: Turbidity of syringes after culturing for 7 days at 37 degrees C was used as the criterion for possible bacterial contamination, which was proved with subsequent Gram staining . MEASUREMENTS AND MAIN RESULTS: A median contamination rate of 22% (range, 7% to 44%) was observed for the syringes prepared from 10-mL ampules by intensive care unit nurses, compared with only 1% for the syringes prepared from ampules by technicians (p <.001) . In >75% of all contaminated syringes, Gram-positive cocci were identified . At least 12% of all prepared syringes proved to be contaminated with staphylococci species . The contamination rate of syringes prepared from vials was much lower: 2% in the intensive care unit and 0% at the department of clinical pharmacy . CONCLUSIONS: In the intensive care unit, standard procedures for preparing syringes for intravenous administration of drugs lack vigorous aseptic precautions, leading to a high contamination rate of the infusate . This risk is increased when ampules instead of 50 mL-vials are used to prepare the syringes. Nutr Hosp, 2002 Jan-Feb, 17(1), 15 - 21 {Long-term follow-up of home parenteral nutrition at a general hospital: complications and quality of life}; Cuerda C et al.; OBJECTIVE: The present study assesses the septic, thrombotic and hepatic complications and also the quality of life of patients with home parenteral nutrition (NPD in its Spanish acronym) . METHODS: The case histories of patients treated with NPD at our hospital were collected for the period January, 1986, to January, 2001 . We had 20 patients, mean age 48 +/- 14 years (mean +/- SD), with a total catheterization time of 16,292 days . The main causes of intestinal failure were mesenteric ischaemia and radical enteritis . Quality of life was assessed by means of the Sickness Impact Profile (SIP) and the Karnofsky Index (KI) . RESULTS: We found a catheter infection rate of 0.42 per 100 days of catheter, mostly produced by coagulase-negative staphylococci (63%) . In 30% of the cases, it was necessary to remove the catheter, including in all cases produced by Candida . Cather occlusion rate was 0.05 per 100 days . Five patients presented an increase in transaminases and alkaline phosphatase, and one female patient developed cholestasis . Two patients died of NPD-related causes . The results of the quality of life analysis showed it was moderately affected, with SIP scores of 14.6 +/- 3.5 and KI results of 70 +/- 20 (mean +/- SD) . CONCLUSIONS: Home parenteral nutrition allows a greater survival in cases of intestinal failure . The most frequent complication is an infection of the catheter . The results of the quality of life analysis in our series reveal it was moderately affected in most cases. J Chemother, 2001 Nov, 13 Spec No 1(1), 48 - 53 Antibiotic prophylaxis in orthopedic prosthetic surgery; de Lalla F; Staphylococci account for approximately 70% of postoperative infections in orthopedic prosthetic surgery, with the leading organism being Staphylococcus epidermidis . Therefore, the antibiotics most widely used for prophylaxis are cefazolin, cefamandole and cefuroxime, by virtue of their excellent activity against these pathogens . However, methicillin-resistant coagulase-positive and -negative staphylococci are increasingly being reported as the causative agents of postoperative infection in clean prosthetic surgery, therefore prompting the use of glycopeptide antibiotics (vancomycin and teicoplanin) in the prophylaxis for total joint replacement, particularly in hospitals in which there is high methicillin-resistance among these pathogens . Furthermore, in total primary knee replacement (TKR) performed with tourniquet application, regional prophylaxis with teicoplanin (i.e . the injection of teicoplanin into a foot vein of the leg to be operated on after occlusion of the systemic circulation by inflating the tourniquet) has been shown to be a safe and valuable prophylactic technique . Apart from providing teicoplanin concentrations in the operative field higher than those achievable by conventional systemic prophylaxis (i.e . by injecting the antibiotic into an arm vein before application of the tourniquet), regional prophylaxis was found in a prospective, controlled, open study on patients undergoing primary monolateral or bilateral TKR to result in a rate of postoperative infection similar to those achievable with conventional prophylactic regimens. J Chemother, 2001 Nov, 13 Spec No 1(1), 224 - 33 Catheter-related infections: diagnosis and intravascular treatment; Bouza E et al.; The diagnosis of catheter-related infections relies on the presence of clinical manifestations of infection and the evidence of colonization of the catheter tip by bacteria, mycobacteria or fungi . The reference method to confirm the latter requires the withdrawal of the catheter for culturing, which frequently turns out to be inconvenient, unnecessary and costly . New methods try to avoid these inconveniences and to assess the presence of tip colonization without withdrawal . One of these methods uses quantitative blood cultures with a jump (> or = 5) in colony counts between blood obtained from the catheter lumen and simultaneously from a peripheral vein . It has a high sensitivity (>80%) and specificity (94%-100%) but is cumbersome and requires both an easy backflow of blood in the catheter and the existence of bacteremia . Cytocentrifugation and acridine orange staining of blood withdrawn from an infected catheter lumen has a sensitivity and a specificity of over 90% for the diagnosis of tip colonization . "Superficial cultures" consist in the semiquantitative culture of the hub, of the skin surrounding the catheter entrance and of the first (1 cm) subcutaneous portion of the catheter after swabbing . Sensitivity of this method is >90% and specificity is >80%, and positive and negative predictive values for catheters (considering together those with and without clinical data of infection) are 66% and 97%, respectively . Endoluminal brushing has turned out to be an impractical and unreliable procedure, at least in our experience . New methods based on the speed of bacterial growth to detectable levels of microorganisms in conventional blood cultures are a new and interesting way of assessing catheter-related infections . Besides, as use of antimicrobial-coated catheters becomes more prevalent, the existing definitions of catheter colonization and catheter-related infection may need to be modified, because such coatings may lead to false-negative culture results . Many catheter infections, diagnosed without catheter withdrawal, can be handled nowadays with the so-called "antibiotic lock-in technique", which consists in locking the infected catheter lumen with a solution containing antibiotics . A high proportion of infected catheters, mainly those with coagulase-negative staphylococci, can be maintained in place and sterilized with this technique, including catheters in patients with therapeutic failure after receiving conventional intravenous antibiotic therapy . New diagnostic and therapeutic techniques may avoid the unnecessary withdrawal of thousands of efficient, difficult to replace and expensive intravascular lines. J Chemother, 2001 Nov, 13 Spec No 1(1), 218 - 23 De-escalation antimicrobial chemotherapy in critically III patients: pros and cons; Antonelli M et al.; In spite of advances in critical care, nosocomial infections still have a considerable impact on Intensive Care Unit (ICU) and hospital length of stay, mortality and costs . Several authors suggest that antibiotic therapy should be instituted as soon as sepsis is suspected in critically patients . Over the last two decades the rates of occurrence for pathogens have significantly changed under selective pressure from broad-spectrum antimicrobial therapy . Shifts from predominance of gram-negative to gram-positive organisms and outbreaks of resistant pathogens address the need for appropriate empirical regimens . Agents such as ceftazidime, imipenem and, more recently, meropenem and tazobactam have been used successfully as monotherapy . Two different clinical trials have reported that meropenem monotherapy is significantly more effective than ceftazidime-based therapy . Because of the outbreak of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis, some investigators suggest adding a glycopeptide to beta-lactamase inhibitor and carbapenem as initial empirical therapy . Such a regimen should be administered before definitive proof of infections and until the results of microbial investigation are available (de-escalation antimicrobial chemotherapy) . On the other hand, several authors do not recommend glycopeptide administration in an attempt to limit nosocomial outbreaks of vancomycin-resistant enterococci (VRE) and staphylococci (VRS) and to avoid secondary drawbacks, such as nephrotoxicity and ototoxicity . De-escalation antimicrobial chemotherapy should be tailored to critically ill patients according to their clinical status, severity of illness and suspicion of sepsis or nosocomial pneumonia. Diagn Microbiol Infect Dis, 2002 Mar, 42(3), 205 - 12 Simultaneous detection of the mecA and ileS-2 genes in coagulase-negative staphylococci isolated from Brazilian hospitals by multiplex PCR; Ferreira RB et al.; Coagulase-negative Staphylococcus spp . (CNS) has been associated with primary bloodstream infections and implanted medical devices . Its importance is increasing due to the acquisition of resistance to oxacillin (Oxa) and, recently, resistance to mupirocin (Mup) . Mupirocin, a topical antimicrobial, has been used in the prevention of staphylococci catheter colonization . Susceptibility to Oxa and Mup was analyzed by different testing methods in clinical CNS isolates . Among 112 CNS strains, 69 (61.6%) were Oxa(R) by the disk diffusion (DD) method and 72 (64.2%) grew on the oxacillin agar screen plate . S . epidermidis and S . haemolyticus presented high rates of oxacillin resistance, 75.4% and 96.1%, respectively . Twenty four (21.4%) strains were Mup(R) by the DD test and 21 of them (87.5%) were identified as S . epidermidis . The detection of the mecA and ileS-2 genes, determined by multiplex-PCR, showed that 72 (64.2%) CNS strains possessed the mecA gene, while 16 (14.3%) possessed the ileS-2 gene . Fifteen of these strains presented the two resistance genes simultaneously . The isolates containing the ileS-2 gene presented a minimum inhibitory concentration (MIC) >1024 microg/mL in the E-test, while low-level mupirocin resistance (MICs of 12-16 microg/mL) was observed in those strains without ileS-2 . The resistances to high and low levels of mupirocin could not be distinguished when the DD test was used . The analysis of the Mup(R) S . epidermidis strains by Pulsed Field Gel Electrophoresis showed that 17 (80.9%) strains belonged to one of two patterns (A and B), which have been shown to be prevalent in hospitals in Rio de Janeiro . This report showed that the PCR method for detection of oxacillin and mupirocin resistance in CNS is necessary to determine accurate rates of these resistance, and will can help in the staphylococcal infections prevention and control policies in Brazil. Scand J Infect Dis, 2002, 34(2), 122 - 6 Synercid plus vancomycin for the treatment of severe methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci infections: evaluation of 5 cases; Sgarabotto D et al.; Synercid (quinupristin/dalfopristin), the first semi-synthetic injectable streptogramin, is a promising alternative to glycopeptides against many Gram-positive multiresistant bacteria . Vancomycin is still considered an effective agent for the treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections but therapeutic failures with glycopeptides have been observed, even for the treatment of infections caused by S . aureus strains sensitive to vancomycin . Synercid, in combination with a glycopeptide, may address this problem without causing significant side effects due to the different toxicity patterns of the 2 antimicrobials . This study reports our experience with the combination of Synercid and vancomycin in 5 patients with severe infection caused by MRSA or methicillin-resistant coagulase-negative Staphylococcus. J Hosp Infect, 2001 Dec, 49 Suppl A, S3 - 11 Antimicrobial susceptibility of Gram-positive bacteria: what's current, what's anticipated? Johnson AP, Livermore DM, Tillotson GS. Changing patterns of pathogens and antibiotic susceptibility present clinicians with difficult choices for antimicrobial prescribing . In particular, multiresistant staphylococci, enterococci and pneumococci present problems in many settings . The number of predictably active antimicrobials is decreasing in many centres, with significant consequences for both patients and society as a whole . New antimicrobial options have been few in recent years and several promising quinolones have been compromised by formulation and/or toxicity issues . Nevertheless, the recent introduction of linezolid and quinupristin/dalfopristin provides clinicians with valuable new options against Gram-positive cocci . These options should further increase with the likely introduction of daptomycin, oritavancin and tigilcycline . A range of surveillance programmes helps monitor the ever-changing patterns of resistance and thus guides clinicians in their empirical prescribing . Empirical use of powerful newer agents may be justifiable in seriously ill patients in those settings, units and countries where there is a substantial background rate of resistance. J Clin Microbiol, 2002 Apr, 40(4), 1514 - 7 Evaluation of a triplex PCR assay to discriminate Staphylococcus aureus from coagulase-negative Staphylococci and determine methicillin resistance from blood cultures; Maes N et al.; A triplex PCR targeting the 16S rRNA, mecA, and nuc genes was developed for identification of staphylococci and detection of methicillin resistance . After validation of the assay with a collection of strains of staphylococci and enterococci (n = 169), the assay was evaluated with cultures of blood with gram-positive cocci from 40 patients . Accurate results were obtained for 59 (98%) of 61 cultures within 6 h of growth detection. J Am Coll Cardiol, 2002 Apr 3, 39(7), 1204 - 11 Periannular extension of infective endocarditis; Graupner C et al.; OBJECTIVES: This prospective study was designed to assess the current clinical course, risk factors, microbiologic profile and echocardiographic findings of patients with left-sided endocarditis and perivalvular complications . BACKGROUND: Periannular complications worsen the prognosis of patients with endocarditis . The relation between these complications and the clinical and microbiologic data has not been clearly defined . METHODS: In this clinical cohort study, 211 patients with left-sided endocarditis, according to the Duke criteria, were prospectively recruited . All patients underwent conventional and transesophageal echocardiography . The mean follow-up interval was 151 days . RESULTS: Perivalvular complications were detected in 78 patients (37%) . The incidence of periannular extension of infection in native and prosthetic valves was 29% and 55%, respectively . The presence of prosthesis (relative risk {RR} 1.88, 95% confidence interval {CI} 1.35 to 2.64) and previous endocarditis (RR 1.78, 95% CI 1.16 to 2.7) were the only pre-existing heart conditions associated with perivalvular complications . Aortic infection (RR 1.8, 95% CI 1.23 to 2.66) and the development of atrioventricular (AV) block (RR 2.55, 95% CI 1.91 to 3.41) were related with the existence of these complications . Coagulase-negative staphylococci were very common in patients with perivalvular complications (RR 1.77, 95% CI 1.21 to 2.59), and small vegetations were more frequent in these patients (RR l.45, 95% CI 0.95 to 2.22) . An operation was more frequently performed in patients with perivalvular complications, but mortality was similar in patients with and without these complications . CONCLUSIONS: Aortic infection, prosthetic endocarditis, new AV block and coagulase-negative staphylococci were independent risk factors of periannular complications . The period between symptom onset and diagnosis, the incidence of pericardial effusion and persistent signs of infection were similar between patients with and without perivalvular complications . Patients with perivalvular complications did not demonstrate a difference in the presence or size of vegetations or the frequency of embolism . An operation was more frequently performed in these patients, but mortality was similar in both groups. Mikrobiologiia, 2002 Jan-Feb, 71(1), 23 - 9 {The role of bacterial growth autoregulators (alkyl hydroxybenzenes) in the response of staphylococci to stresses}; Il'inskaia ON et al.; The investigation of the response of a batch culture of Staphylococcus aureus to exogenous alkyl-substituted hydroxybenzenes (AHBs), chemical analogues of anabiosis autoinducers, showed that C1-AHB at concentrations from 5 microM to 1.5 mM did not influence the culture growth, whereas the more hydrophobic C6-AHB inhibited it at concentrations of 0.5 mM and higher . Either of the AHBs drastically enhanced the phenotypic dissociation of staphylococcal cultures, which manifested itself in an increase in the fraction of cells producing small nonhemolyzing colonies of G type when plated on solid media with erythrocytes . In a submerged staphylococcal culture, the relative number of cells producing G-type colonies varied from 10 to 90%, depending on the concentration of the AHB added . The growth of S . aureus in the presence of AHBs also enhanced cell tolerance to heat shock (heating at 45 or 60 degrees C for 10 min) . The role of AHBs, which are structural modifiers of membranes and possess chaperone activity, in the mechanisms responsible for cell tolerance and phenotypic dissociation of microbial populations is discussed. Presse Med, 2002 Feb 23, 31(7), 297 - 301 {Synercid emergency prescription program . The French experience}; Garrigues B; OBJECTIVE: An emergency-use program forof Synercid (quinupristin/dalfopristin, Q/D) has been set up following the occurrence of Gram-positive infections with no therapeutic alternatives to the available antibiotic arsenal . METHODS: The experience in France is based on a collective of 88 infections analysed in 74 patients . The most frequent clinical indications were: central catheter-related bacteremia, bone and joint infection, endocarditis and, intraabdominal infection . The most frequently causative pathogens were: S . aureus (n = 26, including 24/26 meticillin-resistant), coagulase negative staphylococci (n = 28, including 24/28 meticillin-resistant), enterococci (n = 15), and others (n = 5) . Q/D was administered most frequently by central venous infusion, 3 times a day (68/74 patients); the mean and median dose per infusion was 7.4 mg/kg and the mean duration of treatment was 15.6 days . A combined antibiotic therapy was used in 70/74 patients (a glycopeptide in 41/54 staphylococcal infections) . RESULTS: A Clinical success at the end of treatment was obtained in 40/74 patients (54%; CI 42.1%-65.7%) {the analysis included 25 patients (34%) with an indeterminate clinical response, categorized as failures} et and 39/73 patients (53%) at the follow-up {including 22 deaths (30%), categorized as failure at the follow-up} . The end-of-treatment success rate in patients with staphylococcal and enterococcal infections was respectively 30/54 (56%) and 8/15 (53%) . The safety analysis indicated that 24/74 patients presented at least one treatment-related intercurrent event (possible or probable relationship), the most frequent ones being digestive disordersturbances, signs of venous intolerance, or diffuse or muscular pain . CONCLUSION: Q/D has demonstrated a therapeutic potential in a variety of Gram-positive infections (staphylococcal and enterococcal) in patients with no therapeutic antibiotic alternative, and the type of the intercurrent events reported was consistent with those expected ones in this population of seriously ill patients. Antimicrob Agents Chemother, 2002 Apr, 46(4), 971 - 6 In vitro and in vivo activities of a novel cephalosporin, BMS-247243, against methicillin-resistant and -susceptible staphylococci; Fung-Tomc JC et al.; The recent emergence of methicillin-resistant