J Nippon Med Sch, 2001 Jun, 68(3), 222 - 32 {Cloning of group A streptococcal pyrogenic exotoxin-B gene and its recombinant protein expression in culture supernatant}; Watanabe Y; Streptococcal pyrogenic exotoxin B, a conserved cysteine protease (SPE B/SCP) released by group A Streptococcus (GAS) strains, is considered to be an important virulence factor of this bacterium . This paper reports the cloning of gene encoding SPE B/SCP . For production of recombinant SPE B/SCP (rSPE B/SCP), the primers specific for the SPE B/SCP gene (spe b) were designed based on its nucleotide sequence . Polymerase chain reaction (PCR) was performed with the genomic DNA of GAS strain NZ131 as a template . The amplified PCR products were purified and cloned into the pBluescript II SK(+) plasmid vector . The vector was transformed into Escherichia coli (E . coli) JM109 . The rSPE B/SCP and its recombinant proenzyme (rzym) were secreted in the culture supernate of the transformant . The rSPE B/SCP was purified from the supernatant by sequential chromatography on DEAE-Sepharose, matrix gel Red A and Sephadex G-50 columns . The purified rzym and rSPE B/SCP, respectively, gave a single band with a molecular weight approximately 40 kDa and 27 kDa on SDS-polyacrylamide gel electrophoresis, and reacted with anti-SPE B/SCP antibodies in Western Blot analysis . This is the first report in which rSPE B/SCP was obtained from the culture supernate of the transformant. J Leukoc Biol, 2001 Jun, 69(6), 921 - 7 Superantigen antagonist blocks Th1 cytokine gene induction and lethal shock; Arad G et al.; Bacterial superantigens trigger an excessive, Th1-cytokine response leading to toxic shock . We designed a peptide antagonist that inhibits SEB-induced expression of human genes for IL-2, IFN-gamma, and TNF-beta, cytokines that mediate shock . The peptide antagonist shows homology to a beta-strand-hinge-alpha-helix domain that is conserved structurally in superantigens produced by Staphylococcus aureus and Streptococcus pyogenes yet remote from known binding sites for the major histocompatibility class II molecule and T-cell receptor . For Th1-cell activation, superantigens depend on this domain . The peptide protected mice against lethal challenge with SEB or SEA . Moreover, it rescued mice undergoing toxic shock . Surviving mice rapidly developed broad-spectrum, protective immunity, which rendered them resistant to further lethal challenges with different staphylococcal and streptococcal superantigens . Thus, the lethal effect of superantigens, mediated by Th1 cytokines, can be blocked with a peptide antagonist that inhibits their action at the top of the toxicity cascade, before activation of T cells takes place. J Biol Chem, 2001 Aug 24, 276(34), 31494 - 501 Epub 2001 Jun 14. Kinetics of beta-lactam interactions with penicillin-susceptible and -resistant penicillin-binding protein 2x proteins from Streptococcus pneumoniae . Involvement of acylation and deacylation in beta-lactam resistance; Lu WP et al.; Kinetic interactions of beta-lactam antibiotics such as penicillin-G and cefotaxime with normal, penicillin-susceptible PBP2x from Streptococcus pneumoniae and a penicillin-resistant PBP2x (PBP2x(R)) from a resistant clinical isolate (CS109) of the bacterium have been extensively characterized using electrospray mass spectrometry coupled with a fast reaction (quench flow) technique . Kinetic evidence for a two-step acylation of PBP2x by penicillin-G has been demonstrated, and the dissociation constant, K(d) of 0.9 mm, and the acylation rate constant, k(2) of 180 s(-1), have been determined for the first time . The millimolar range K(d) implies that the beta-lactam fits to the active site pocket of the penicillin-sensitive PBP rather poorly, whereas the extremely fast k(2) value indicates that this step contributes most of the binding affinity of the beta-lactam . The values of K(d) (4 mm) and k(2) (0.56 s(-1)) were also determined for PBP2x(R) . The combined value of k(2)/K(d), known as overall binding efficiency, for PBP2x(R) (137 m(-1) s(-1)) was over 1000-fold slower than that for PBP2x (200,000 m(-1) s(-1)), indicating that a major part is played by the acylation steps in penicillin resistance . Most of the decreased binding efficiency of PBP2x(R) comes from the decreased ( approximately 300-fold) k(2) . Kinetic studies of cefotaxime acylation of the two PBP2x proteins confirmed all of the above findings . Deacylation rate constants (k(3)) for the third step of the interactions were determined to be 8 x 10(-6) s(-1) for penicilloyl-PBP2x and 5.7 x 10(-4) s(-1) for penicilloyl-PBP2x(R), corresponding to over 70-fold increase of the deacylation rate for the resistant PBP2x(R) . Similarly, over 80-fold enhancement of the deacylation rate was found for cefotaxime-PBP2x(R) complex (k(3) = 3 x 10(-4) s(-1)) as compared with that of cefotaxime-PBP2x complex (3.5 x 10(-6) s(-1)) . This is the first time that such a significant increase of k(3) values was found for a beta-lactam-resistant penicillin-binding protein . These data indicate that the deacylation step also plays a role, which is much more important than previously thought, in PBP2x(R) resistance to beta-lactams. Antimicrob Agents Chemother, 2001 Jul, 45(7), 2169 - 72 Activity of LY333328 in experimental meningitis caused by a Streptococcus pneumoniae strain susceptible to penicillin; Gerber J et al.; In a rabbit model of Streptococcus pneumoniae meningitis single doses of 10 and 2.5 mg of the glycopeptide LY333328 per kg of body weight reduced bacterial titers in cerebrospinal fluid (CSF) almost as rapidly as ceftriaxone at 10 mg/kg/h (changes in log CFU, -0.29 +/- 0.21 and -0.26 +/- 0.22 versus -0.34 +/- 0.15/ml/h) . A dose of 1 mg/kg was bacteriostatic (change in log CFU, 0.01 +/- 0.11/ml/h) . In two animals receiving LY333328 at a dose of 40 mg/kg the bacterial titers were reduced by 0.54 and 0.51 log CFU/ml/h . The penetration of CSF by LY333328 was 1 to 5% . The concentrations of lipoteichoic and teichoic acids in CSF and neuronal damage were similar in ceftriaxone- and LY333328-treated animals. Antimicrob Agents Chemother, 2001 Jul, 45(7), 2163 - 8 Comparative in vitro activity of ABT-773, a novel antibacterial ketolide; Nilius AM et al.; The in vitro activities of ABT-773, erythromycin, clarithromycin, and azithromycin were compared . ABT-773 was the most active compound against macrolide-susceptible Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Staphylococcus epidermidis, Listeria monocytogenes, and Enterococcus spp . and multidrug-resistant Streptococcus pneumoniae . It also had good activity against gram-negative and atypical respiratory tract pathogens and Helicobacter pylori. Antimicrob Agents Chemother, 2001 Jul, 45(7), 2147 - 50 Macrolide-resistant Streptococcus pneumoniae in Canada during 1998-1999: prevalence of mef(A) and erm(B) and susceptibilities to ketolides; Hoban DJ et al.; In this study (1998-1999), we collected 215 macrolide-resistant Streptococcus pneumoniae isolates from an ongoing Canadian Respiratory Organism Surveillance Study involving 23 centers representing all regions of Canada . The prevalence of erythromycin-resistant S . pneumoniae was 8% (215 of 2,688) . Of the 215 isolates, 48.8% (105 of 215) were PCR positive for mef(A) and 46.5% (100 of 215) were PCR positive for erm(B) . The ketolides telithromycin and ABT-773 demonstrated excellent activity against both mef(A) (MIC for 90% of strains {MIC(90)}, 0.06 and 0.03 microg/ml, respectively) and erm(B) (MIC(90), 0.06 and 0.03 microg/ml, respectively) strains of S . pneumoniae. Antimicrob Agents Chemother, 2001 Jul, 45(7), 2092 - 7 Pharmacodynamic assessment of gatifloxacin against Streptococcus pneumoniae; Mattoes HM et al.; The pharmacodynamic parameters of peak serum drug concentration/MIC (peak/MIC) ratio and the area under the curve (AUC)/MIC ratio have been used to characterize in vivo drug exposure and its relationship to bacterial killing for the fluoroquinolones . Our study objectives were to describe the pharmacodynamic relationship between gatifloxacin exposure and outcome as assessed by bacterial density and survival in an immunocompromised murine thigh model of pneumococcal infection and to assess the relationship between drug exposure and these outcomes in an immunocompetent host . ICR mice were rendered neutropenic, and thigh infection was induced by intramuscular administration of 0.1 ml of 10(5) to 10(7) CFU of Streptococcus pneumoniae/ml . Mice received 1 to 5 mg of uranyl nitrate/kg of body weight at day -3 and were randomized to receive 10 to 80 mg of gatifloxacin/kg every 6 to 24 h orally, starting at 2 h postinoculation . Bacterial density studies were completed 24 h after initiation of therapy, and survival was assessed after 4 days of treatment . MICs for clinical isolates (n = 8) ranged from 0.25 to 1.0 microg/ml . Correlations were assessed between the change in bacterial density, as well as survival, and the AUC/MIC ratio, peak/MIC ratio, and the duration of time that serum drug concentration remained above the MIC . The best predictor of bacterial response was the AUC/MIC ratio for both outcome measures . There was greater efficacy, as measured by a decrease in log change in CFU as well as by survival data, in the immunocompetent mice compared to the immunocompromised mice . These data demonstrate (i) the appropriateness of the AUC/MIC ratio as a dynamic predictor of response to pneumococcal infection for the fluoroquinolones, (ii) that gatifloxacin AUC/MIC ratios of 30 to 40 appear to optimize bactericidal activity and survival in this model, and (iii) that immunocompetency of the host plays a role in efficacy. Antimicrob Agents Chemother, 2001 Jul, 45(7), 2075 - 81 All detectable high-molecular-mass penicillin-binding proteins are modified in a high-level beta-lactam-resistant clinical isolate of Streptococcus mitis; Amoroso A et al.; All detectable high-molecular-mass penicillin-binding proteins (HMM PBPs) are altered in a clinical isolate of Streptococcus mitis for which the beta-lactam MICs are increased from those previously reported in our region (cefotaxime MIC, 64 microg/ml) . These proteins were hardly detected at concentrations that saturate all PBPs in clinical isolates and showed, after densitometric analysis, 50-fold-lower radiotracer binding . Resistance was related to mosaic structure in all HMM PBP-coding genes, where critical region replacement was complemented not only by substitutions already reported for the closely related Streptococcus pneumoniae but also by other specific replacements that are presumably close to the active-site serine . Mosaic structure was also presumed in a pbp1a-sensitive strain used for comparison, confirming that these structures do not unambiguously imply, by themselves, detectable critical changes in the kinetic properties of these proteins. Rapid Commun Mass Spectrom, 2001, 15(12), 915 - 9 Determination of gatifloxacin in human plasma by liquid chromatography/electrospray tandem mass spectrometry; Vishwanathan K et al.; Gatifloxacin is an advanced-generation, 8-methoxyfluoroquinolone that is active against a broad spectrum of pathogens, including antiobiotic resistant Streptococcus pneumoniae . Development of a rapid, sensitive and selective method for the determination of gatifloxacin in human plasma is essential for understanding the pharmacokinetics of the drug when administered orally or intravenously . Solid phase extraction (SPE) using Oasis HLB was used to extract gatifloxacin and the internal standard ciprofloxacin from plasma . A method based on liquid chromatography/electrospray tandem mass spectrometry (LC/ESI-MS/MS) was developed and validated to quantitate gatifloxacin in human plasma . The precursor and major product ions of the analyte were monitored on a triple quadrupole mass spectrometer with positive ion electrospray ionization (ESI) in the multiple reaction monitoring (MRM) mode . Mechanisms for the formation of collision-induced dissociation products of gatifloxacin are proposed . Linear calibration curves were generated from 10--1000 ng/mL with coefficients of determination greater than 0.99 . The interday and intraday precision (%RSD) was less than 6.0% and accuracy (%error) was less than 5.4% for gatifloxacin . The limit of detection (LOD) for the method was 500 pg/mL based on a signal-to-noise ratio of 3 . Chest, 2001 Jun, 119(6), 1717 - 23 Influenza pneumonia: a descriptive study; Oliveira EC et al.; OBJECTIVE: To describe the clinical features and complications of patients hospitalized with influenza during the 1999-2000 influenza season . METHODS: We reviewed all cases of patients with influenza admitted to a large metropolitan referral hospital during the 1999-2000 season . RESULTS: Thirty-five adult patients (15 men and 20 women) tested positive for influenza A by direct enzyme immunoassay . A fourfold to sevenfold increase in the number of influenza cases was observed over previous years . Most patients had serious comorbid illnesses (88%), such as diabetes and chronic respiratory and heart disease . Seventeen patients developed pneumonia; these patients tended to be older (mean +/- SD, 63 +/- 13 years vs 51 +/- 19 years, respectively; p = 0.04) and had a higher incidence of chronic lung disease (41% vs 6%, respectively; p = 0.02) than the patients without pneumonia . Shortness of breath was the only symptom that distinguished patients with pneumonia from those with an upper respiratory tract illness alone . Antiviral treatment was begun 4 +/- 3 days from initiation of symptoms in patients with pneumonia and consisted of oseltamivir (35.2%), rimantadine (5.8%), or a combination of both (17.6%) . Respiratory and/or blood culture results were positive in five patients (29%), Staphylococcus aureus was isolated in five patients, and Streptococcus pneumoniae was isolated in one patient . Ten of the patients with pneumonia (58.8%) were admitted to the ICU, and 5 patients (29%) died . The length of ICU stay and mechanical ventilation were 28 +/- 26 days and 21.5 +/- 20.5 days, respectively . Death in most pneumonia patients was attributed to respiratory failure . CONCLUSION: The recognized number of hospital admissions for influenza increased fourfold to sevenfold over previous years, most likely due to the implementation of rapid diagnostic tests for influenza . Patients with signs and symptoms of influenza and shortness of breath should undergo chest radiography . Hospitalization of patients with influenza pneumonia occurred in both previously healthy and immunocompromised patients and had a high mortality . S aureus was the most common bacterial isolate in patients with influenza pneumonia . Empiric antibiotics with staphylococcal activity should be used pending culture results in patients with influenza pneumonia . The effectiveness of oseltamivir and rimantadine in treating patients with influenza pneumonia remains to be determined. J Am Geriatr Soc, 2001 May, 49(5), 557 - 63 Aspiration pneumonia: dental and oral risk factors in an older veteran population; Terpenning MS et al.; OBJECTIVES: To investigate the importance of medical and dental factors in aspiration pneumonia in an older veteran population . DESIGN: Prospective enrollment of subjects with retrospective analysis of data . SETTING: Department of Veterans Affairs outpatient clinic, inpatient ward, and nursing home . PARTICIPANTS: 358 veterans age 55 and older; 50 subjects with aspiration pneumonia . MEASUREMENTS: Demographic and medical data; functional status; health-related behaviors; dental care utilization; personal oral hygiene; comprehensive dental examination; salivary assays including IgA antibodies; and cultures of saliva, throat, and dental plaques . RESULTS: Two logistic regression models produced estimates of significant risk factors . One model using dentate patients included: requiring help with feeding (odds ratio (OR) = 13.9), chronic obstructive pulmonary disease (COPD) (OR = 4.7), diabetes mellitus (OR = 3.5), number of decayed teeth (OR = 1.2), number of functional dental units (OR = 1.2), presence of important organisms for decay, Streptococcus sobrinus in saliva (OR = 6.2), and periodontal disease, Porphyromonous gingivalis in dental plaque (OR = 4.2), and Staphylococcus aureus presence in saliva (OR = 7.4) . The second model, containing both dentate and edentulous patients included: requiring help with feeding (OR = 4.7), COPD (OR = 2.5), diabetes mellitus (OR = 1.7), and presence of S . aureus in saliva (OR = 8.3) . CONCLUSION: This study supports the significance of oral and dental factors while controlling for established medical risk factors in aspiration pneumonia incidence. Kurume Med J, 2001, 48(1), 1 - 8 Clinical and laboratory evaluation of penicillin resistant Streptococcus pneumoniae in relation to the mutations of pbp 1a, pbp2b and pbp2x; Tanoue S; A total of 210 isolates of Streptococcus pneumoniae (S . pneumoniae) were selected randomly to examine drug susceptibility which were obtained from out- and in-patients who visited Kurume University Hospital and other affiliated hospitals through May 1998 to September 1999 . The prevalence of penicillin resistant S . pneumoniae in this study was 39.5% and was compatible with those of previous reports in Japan . From the clinical aspects, the resistant strains of S . pneumoniae have been shown not to be so highly virulent comparing with sensitive strains that only few severe or mortal cases had been reported . Carbapenems, glycopeptides, and fluoroquinolones were shown to be highly active against penicillin-resistant S . pneumoniae strains as evidenced by the low minimum inhibitory concentrations (MICs) though LVFX showed 4 micrograms/ml or higher MICs against some strains . Regarding to the mechanisms of penicillin resistance, penicillin binding proteins coding gene (pbp1a, pbp2x and pbp2b) mutations that cause modifications in these proteins were also examined for all isolates . The multivariate analysis showed statistically significant correlation between higher MIC of penicillin and cephem and pbp1a mutation while no significant contribution of pbp2x and pbp2b to the resistance was demonstrated . Additionally, no significant correlation between pbp mutation and MIC of carbapenem was observed . Furthermore, there were two highly penicillin resistant S . pneumoniae strains with no pbp mutations . Thus the pbp mutations alone were not responsible for the elevation of MIC all beta-lactams . Nevertheless the pbp mutations were detected in advance of actual MIC elevations by inducement experiment in vitro . It indicated that penicillin resistance might be detected earlier than conventional methods . In previous reports some other responsible genes for penicillin resistance were demonstrated . Therefore, it might be possible to presume exact values of MICs of beta-lactams against resistant strains of S . pneumoniae by detecting still unknown genes other than pbps. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2001 Jun, 91(6), 643 - 8 Evaluation of oral antimicrobial agent levels in tooth extraction sites; Yoshii T et al.; OBJECTIVE: The purpose of this study was to evaluate various oral antimicrobial agent levels in tooth extraction sites . STUDY DESIGN: The concentration of dental alveolar blood in extraction wounds after the oral administration of talampicillin (500 mg), cefaclor (500 mg), cefteram pivoxil (200 mg), cefuroxime axetil (250 mg), cefdinir (200 mg), and ofloxacin (100 mg) was determined in 338 patients and was assessed on the basis of its antimicrobial activity against Streptococcus isolated in odontogenic infections . RESULTS: The percentage of patients whose concentrations exceeded the minimum inhibitory concentration for 90% of Streptococcus was 62.5% to 100% for talampicillin at 30 to 360 minutes, 0% to 12.5% for cefaclor at 30 to 360 minutes, 18.2% to 100% for cefteram pivoxil at 30 to 480 minutes, 50% to 100% for cefuroxime axetil at 30 to 480 minutes, 0% to 50% for cefdinir at 16 to 290 minutes, and 0% to 40% for ofloxacin at 30 to 480 minutes . CONCLUSION: These results indicate that talampicillin, cefteram pivoxil, and cefuroxime axetil have minimum inhibitory concentration levels for 90% of Streptococcus in tooth sockets. Infect Immun, 2001 Jul, 69(7), 4698 - 701 Synthetic polysaccharide type 3-related di-, tri-, and tetrasaccharide-CRM(197) conjugates induce protection against Streptococcus pneumoniae type 3 in mice; Benaissa-Trouw B et al.; Di-, tri-, and tetrasaccharides, synthesized according to the chemical structure of pneumococcal polysaccharide type 3 (PS3), were coupled to the cross-reactive material (CRM(197)) of modified diphtheria toxin in different molar carbohydrate/protein ratios using the squarate coupling method . To study protective immunity, female BALB/c mice were subcutaneously immunized twice (with a 3-week interval) using the amount of conjugates corresponding to 2.5 microg of oligosaccharide per mouse . The conjugates evoked PS3 binding immunoglobulin G antibodies that lasted for at least 7 weeks after the booster . Immunogenicity was not influenced by the carbohydrate/protein ratio . All mice with PS3-specific antibodies survived the intraperitoneal challenge with Streptococcus pneumoniae type 3 . Therefore, synthetic oligosaccharide-protein conjugates might have potential as vaccines. Infect Immun, 2001 Jul, 69(7), 4647 - 53 Expression and functional properties of the Streptococcus intermedius surface protein antigen I/II; Petersen FC et al.; Streptococcus intermedius is associated with deep-seated purulent infections . In this study, we investigated expression and functional activities of antigen I/II in S . intermedius . The S . intermedius antigen I/II appeared to be cell surface associated, with a molecular mass of approximately 160 kDa . Northern blotting indicated that the S . intermedius NCTC 11324 antigen I/II gene was transcribed as a monocistronic message . Maximum expression was seen during the early exponential phase . Insertional inactivation of the antigen I/II gene resulted in reduced hydrophobicity during early exponential phase, whereas no effect was detected during mid- and late exponential phases . Binding to human fibronectin and laminin was reduced in the isogenic mutant, whereas binding to human collagen types I and IV and to rat collagen type I was not significant for either the wild type or the mutant . Compared to the wild type, the capacity of the isogenic mutant to induce interleukin 8 (IL-8) release by THP-1 monocytic cells was significantly reduced . The results indicate that the S . intermedius antigen I/II is involved in adhesion to human receptors and in IL-8 induction. Mol Microbiol, 2001 May, 40(4), 976 - 90 Repression of virulence genes by phosphorylation-dependent oligomerization of CsrR at target promoters in S . pyogenes; Miller AA et al.; csrRS encodes a two-component regulatory system that represses the transcription of a number of virulence factors in Streptococcus pyogenes, including the hyaluronic acid capsule and pyrogenic exotoxin B . CsrRS-regulated virulence factors have diverse functions during pathogenesis and are differentially expressed throughout growth . This suggests that multiple signals induce CsrRS-mediated gene regulation, or that regulated genes respond differently to CsrR, or both . As a first step in dissecting the csrRS signal transduction pathway, we determined the mechanism by which CsrR mediates the repression of its target promoters . We found that phosphorylated CsrR binds directly to all but one of the promoters of its regulated genes, with different affinities . Phosphorylation of CsrR enhances both oligomerization and DNA binding . We defined the binding site of CsrR at each of the regulated promoters using DNase I and hydroxyl radical footprinting . Based on these results, we propose a model for differential regulation by CsrRS. Can J Microbiol, 2001 May, 47(5), 412 - 6 Iron content of Streptococcus suis and evidence for a dpr homologue; Niven DF et al.; The type strain of Streptococcus suis was investigated for features that might help the organism to tolerate the H2O2 that is produced during growth . Enzyme assays, using soluble extracts, revealed that the type strain, which lacks catalase, lacks NADH peroxidase in both the mid-exponential and stationary phases of the growth cycle . Although iron could not be detected colourimetrically in dense cell suspensions, determination of the cellular iron content following growth to early stationary phase in the presence of 55FeCl3 demonstrated that S . suis does contain iron and hence is incapable of iron exclusion . Gene amplification, using oligonucleotide primers based on dpr of Streptococcus mutans, followed by nucleotide sequencing, revealed in S . suis, the presence of a gene that encodes a Dpr homologue . It is concluded that in S . suis, tolerance of H2O2 is due to iron sequestration by Dpr and the consequent effect of this process on the extent of Fenton chemistry. J Infect Dis, 2001 Jul 1, 184(1), 66 - 72 Epub 2001 Jun 08. Complex relationship between acquisition of beta-lactam resistance and loss of virulence in Streptococcus pneumoniae; Rieux V et al.; In a previous study of a murine peritonitis model, no Streptococcus pneumoniae strains were found that were both clinically penicillin resistant and virulent . This study assessed the relationship between acquired resistance and virulence in single- and double-isogenic penicillin-resistant (Peni-R) mutants obtained by transformation of a virulent penicillin-susceptible recipient strain with pbp2b and pbpX polymerase chain reaction fragments from a Peni-R donor strain . Sequence analysis results of the pbp2b and pbpX alleles from these strains were in keeping with acquired penicillin resistance . The virulence of these strains was significantly reduced, which shows a relationship between beta-lactam resistance and loss of virulence . The phenotype of the 23.2x mutant remained stable after in vivo passage, which suggests that the pbpX gene is involved in growth, whereas virulent revertants of the 23.2b and 23.2b.2x mutants had no change in MIC . Compensatory mutations are implicated in the revival of virulence. J Infect Dis, 2001 Jul 1, 184(1), 56 - 65 Epub 2001 May 31. Structure and dissemination of a chromosomal insertion element encoding macrolide efflux in Streptococcus pneumoniae; Gay K et al.; Macrolide resistance associated with macrolide efflux (mef) has rapidly increased in Streptococcus pneumoniae . We defined the genetic structure and dissemination of a novel mefE-containing chromosomal insertion element . The mefE gene was found on the 5' end of a 5.5- or 5.4-kb insertion designated as the macrolide efflux genetic assembly (mega), which is found in > or =4 distinct sites of the pneumococcal genome . The element was transformable and conferred macrolide resistance to susceptible S . pneumoniae . The first 2 open-reading frames (ORFs) of the element formed an operon composed of mefE and a predicted adenosine triphosphate-binding cassette homologous to msrA . Convergent to this efflux operon were 3 ORFs with homology to stress response genes of Tn5252 . Mega was related to the recently described mefA-containing element Tn1207.1 but lacked the genes necessary for transposition and had unique termini and insertion sites . In metropolitan Atlanta, macrolide resistance due to mega rapidly increased in S . pneumoniae by clonal expansion and horizontally by transformation. Int J Antimicrob Agents, 2001 Jun, 17(6), 465 - 9 Antimicrobial resistance and serotype distribution of Streptococcus pneumoniae isolates from Crete, Greece; Maraki S et al.; Susceptibility to 14 antibiotics was determined for 125 clinical isolates of Streptococcus pneumoniae collected over a 3-year period in Crete, Greece . Twenty-three isolates (18.4%) showed intermediate resistance and 15 (12%) high-level resistance to penicillin . Erythromycin, chloramphenicol, tetracycline, trimethoprim-sulphamethoxazole and sparfloxacin resistance rates were 16.8, 10.4, 19.2, 24.8 and 9.6%, respectively . Multiple resistance was observed in 22 strains . Vancomycin and levofloxacin were the most active agents tested . The most prevalent serotype among penicillin-susceptible pneumococci was 14, followed by 9, 7 and 1, while among penicillin-intermediate or -resistant strains serotype 23 was predominant followed by 19 and 9 . These results show that as well as a high level of penicillin resistance in this region, some strains are also resistant to other antibiotics and may show multi-drug resistance. Vet Ophthalmol, 2000, 3(2-3), 121 - 125 Ulcerative keratitis caused by beta-hemolytic Streptococcus equi in 11 horses; Brooks DE et al.; Purpose To describe 11 clinical cases of ulcerative keratitis in horses associated with beta-hemolytic Streptococcus equi in Florida, USA . METHODS: Retrospective clinical study (1996-99) . RESULTS: Beta-hemolytic Streptococcus equi was cultured from 11 horses with deep ulcers, descemetoceles or iris prolapse (n = 8), a suture abscess found with a penetrating keratoplasty for a stromal abscess (n = 1), and ulceration that developed following keratectomy/irradiation for corneal squamous cell carcinoma (n = 2) . Beta-hemolytic Streptococcus equi subspecies zooepidemicus was found in 10 eyes and subspecies equi in one . Marked signs of uveitis including miosis and hypopyon were present in 8/11 (72.7%) eyes . Keratomalacia was severe in all eyes . The mean diameter of the ulcers associated with beta-hemolytic Streptococcus was 10.2 +/- 6.1 mm . Eight of the eyes required conjunctival flap surgery (four grafts dehisced) and one eye corneal transplantation . Two eyes were treated with medication only . Isolate sensitivity to antibiotics included ampicillin (6/11), bacitracin (11/11), cephalothin (11/11), chloramphenicol (11/11), gentamicin (5/11), polymyxin B (2/11), and tobramycin (1/11) . All isolates were resistant to neomycin . The average healing time was 44.7 +/- 26.7 days . The visual outcome was positive in 8/11 eyes, and the globe retained in 9/11 eyes . CONCLUSIONS: Although Gram-positive bacteria predominate in the normal conjunctival microflora of horses throughout the world, Gram-negative bacteria and fungi are more often isolated from equine ulcers . Beta-hemolytic Streptococcus spp . are associated with a very aggressive ulcerative keratitis with the capability to digest conjunctival graft tissue . Clinical signs are pronounced . Aggressive surgical and intensive medical therapy with topical antibiotics and protease inhibitors is indicated. J Immunother, 2001 May, 24(3), 250 - 256 Meta-analysis of Adjuvant Immunochemotherapy Using OK-432 in Patients With Resected Non-Small-Cell Lung Cancer; Sakamoto J et al.; SUMMARY: The benefits of immunochemotherapy with a penicillin-treated, lyophilized preparation of Streptococcus pyogenes, OK-432 (Picibanil), were reassessed in patients with resected non-small-cell lung cancer through a meta-analysis based on data from 1,520 patients enrolled in 11 randomized clinical trials . All 11 trials were started before 1991, and the subjects had been followed up for at least 5 years after surgery and randomization . In these trials, standard chemotherapy was compared with the same therapy plus OK-432 . The endpoint of interest was overall survival, and analysis was based on intent-to-treat population without patient exclusion . Data were analyzed using the Mantel-Haenszel method . The 5-year survival rate for all eligible patients in the 11 trials was 51.2% in the immunochemotherapy group versus 43.7% in the chemotherapy group . The odds ratio (OR) for overall survival was 0.70 (95% CI = 0.56-0.87, p = 0.0010) . Analysis of four trials in which central randomization was performed also reconfirmed a significantly longer survival time for the immunochemotherapy group (OR = 0.66, 95% CI = 0.44-1.00, p = 0.049) . Based on these results of meta-analysis, it is postulated that postoperative adjuvant immunochemotherapy using OK-432 might improve the survival of patients after resection of non-small-cell lung cancer. J Immunother, 2001 May-Jun, 24(3), 250 - 6 Meta-analysis of adjuvant immunochemotherapy using OK-432 in patients with resected non-small-cell lung cancer; Sakamoto J et al.; The benefits of immunochemotherapy with a penicillin-treated, lyophilized preparation of Streptococcus pyogenes, OK-432 (Picibanil), were reassessed in patients with resected non-small-cell lung cancer through a meta-analysis based on data from 1,520 patients enrolled in 11 randomized clinical trials . All 11 trials were started before 1991, and the subjects had been followed up for at least 5 years after surgery and randomization . In these trials, standard chemotherapy was compared with the same therapy plus OK-432 . The endpoint of interest was overall survival, and analysis was based on intent-to-treat population without patient exclusion . Data were analyzed using the Mantel-Haenszel method . The 5-year survival rate for all eligible patients in the 11 trials was 51.2% in the immunochemotherapy group versus 43.7% in the chemotherapy group . The odds ratio (OR) for overall survival was 0.70 (95% CI = 0.56-0.87, p = 0.0010) . Analysis of four trials in which central randomization was performed also reconfirmed a significantly longer survival time for the immunochemotherapy group (OR = 0.66, 95% CI = 0.44-1.00, p = 0.049) . Based on these results of meta-analysis, it is postulated that postoperative adjuvant immunochemotherapy using OK-432 might improve the survival of patients after resection of non-small-cell lung cancer. Ceska Gynekol, 2000 Dec, 65 Suppl 1, 42 - 6 {Neonatal mortality in the Czech Republic 1998-1999}; Plavka R; OBJECTIVE: To evaluate neonatal mortality rate (NMR) in 1998 and 1999 years in the Czech Republic . DESIGN: Retrospective epidemiological study of all alive newborns born in 1998 and 1999 in the Czech Republic . SETTINGS: 12 perinatological centers of nine regions of Bohemia and Moravia . METHODS: All alive, died, died with congenital defects newborns were registered and results of neonatal mortality rate and specific neonatal mortality rate were calculated . The main causes of death were divided into four groups (intraventricular hemorrhage grade III-IV, infection, acute respiratory failure and others) and evaluated comparatively . In 1999 the NMR of newborns with birth weight below 500 g and their survival were introduced for the first time in the Czech Republic . RESULTS: The fluent decrease of NMR during nineties was stopped in 1999 . Increase of NMR from 2.8@1000 in 1998 to 3.0@1000 in 1999 was mainly caused by arise of specific neonatal mortality rate in newborns weighing > or = 2000 g . Comparing 1998 and 1999 years, two times more these newborns without serious congenital defects died in 1999 (28 vs . 56) . Specific neonatal mortality rate of extremely low birth weight newborns further decreased (359@1000 vs . 279@1000) especially in the newborns with birth weight between 500-749 g (543@1000 vs . 373@1000) . The most frequent main causes of death still has been intraventicular haemorrhage grade III-IV and infection in very low birth weight newborns, and serious congenital defects and infection in newborns weighing > or = 1500 g . The concentration of very low birth weight newborns to perinatological centers by transfer in uterus was 81% in 1998 and 83% in 1999 . The differences in neonatal mortality rates between nine regions of Bohemia and Moravia has been getting equal but has been still great in specific neonatal mortality rate of extremely low birth weight newborns between the best and worst regions (147@1000 vs . 458@1000) . There were registered 19 newborns weighing < or = 500 g surviving more than 24 hours after delivery in the Czech republic . Specific neonatal mortality rate of these newborns was 316@1000 and 527@1000 survived . CONCLUSION: Reserves for further lowering of NMR are improving the care after extremely low birth weight newborns in the regions with below average results and decreasing the mortality of newborns with birth weight > or = 2000 g by introducing of group B streptococcus prophylaxis, improving prenatal diagnostics of serious congenital defects and early and more quality postnatal transport of newborns suffered from acute respiratory failure to centers disposing of the latest methods of treatment. Drugs Aging, 2001, 18(5), 305 - 11 Pneumococcal disease in the elderly: what is preventing vaccine efficacy? Rubins JB, Janoff EN. The effective prevention of Streptococcus pneumoniae infection has a renewed priority in an era in which the emergence of antibacterial-resistant strains has the potential to further compromise efforts to reduce early mortality from invasive pneumococcal infection . Although the 23-valent pneumococcal polysaccharide (PPS) vaccine was approved in the US to prevent respiratory and invasive infection in the elderly and other high-risk populations, the protective efficacy of this vaccine for the growing population of adults aged >65 years remains controversial . The apparent effectiveness of pneumococcal immunisation in clinical studies of elderly adults has varied depending upon whether a reduction in pneumococcal colonisation, pneumonia, bacteraemia or death was used as an outcome . Clinical studies of vaccine efficacy to date suggest that the current pneumococcal vaccine is 56 to 81% effective at preventing invasive pneumococcal infection, and may have additive benefit to influenza vaccine in preventing community-acquired pneumonia, particularly in elderly adults with an increased risk of serious pneumonia requiring hospitalisation . Possible reasons for incomplete protection from pneumococcal infection after immunisation include infection with non-vaccine serotypes, inadequate or ineffective antibody responses, waning of antibody responses, or compromised nonhumoral host defences . Further studies are needed to determine whether: (i) elderly adults who respond poorly to the 23-valent pneumococcal vaccine can be identified prior to immunisation and targeted for study with improved pneumococcal vaccines; (ii) specific nutrient deficiencies can be identified and corrected to improve the immune responsiveness of elderly adults to the PPS vaccine; (iii) newer protein-conjugate or DNA pneumococcal vaccines may be more uniformly immunogenic for elderly adults; and (iv) whether smoking cessation reduces the risk of invasive pneumococcal infection in elderly adults. J Am Soc Echocardiogr, 2001 Jun, 14(6), 644 - 5 Infected left atrial myxoma; Dekkers P et al.; Upon examination, a 40-year-old man was found to have fever, weight loss, and malaise . A blood culture was positive for Streptococcus mutans . Under the suspicion of endocarditis, he was treated with penicillin . Echocardiography revealed a large tumor in the left atrium . After 6 weeks of penicillin treatment, he was transferred to our hospital for excision of the tumor . Pathology revealed a myxoma with fibrin deposits, bacterial colonization, and massive infiltration with neutrophils. Microbiology, 2001 Jun, 147(Pt 6), 1599 - 610 Expression of Streptococcus mutans fimA is iron-responsive and regulated by a DtxR homologue; Spatafora G et al.; Iron uptake, transport and storage in Streptococcus mutans, the principal causative agent of human dental cavities, is unexplored despite early reports in the literature which predict a role for this trace metal in cariogenesis . Experiments in the authors' laboratory revealed several iron-responsive proteins in S . mutans, one of which reacted with a polyclonal antiserum directed against the FimA fimbrial adhesin from Streptococcus parasanguis on Western blots . The results of Western blot and Northern hybridization experiments support an inverse relationship between iron availability and S . mutans fimA expression, and metal ion uptake experiments implicate FimA in S . mutans (55)Fe transport . Cloning of the S . mutans fimA homologue facilitated the construction of a fimA knockout mutant which grew poorly in an iron-limiting medium relative to the wild-type progenitor strain, lending further support to a role for FimA in S . mutans iron transport . The authors also identified and cloned a dtxR-like gene (dlg) located downstream of fimA on the S . mutans chromosome, and noted increased fimA expression in a S . mutans dlg knockout mutant relative to wild-type on RNA spot blots and Western blots . The uptake of (55)Fe, which was also significantly increased in this mutant, was compromised in a fimA/dlg double knockout . These findings are consistent with a role for Dlg in the iron-mediated regulation of fimA, and possibly other S . mutans iron transporters . Finally, the cariogenic potential of the fimA and dlg knockout mutants was not significantly different from that of the wild-type progenitor in a germ-free rat model. J Immunol, 2001 Jun 15, 166(12), 7362 - 9 The differential roles of LFA-1 and Mac-1 in host defense against systemic infection with Streptococcus pneumoniae; Prince JE et al.; Mice deficient in CD18, which lack all four CD11 integrins, have leukocytosis and increased susceptibility to bacterial infection . To determine the effect of deficiencies in LFA-1 (CD11a/CD18) or Mac-1 (CD11b/CD18) on host defense against systemic bacterial infection, knockout mice were inoculated i.p . with Streptococcus pneumoniae . Increased mortality occurred in both LFA-1(-/-) (15 of 17 vs 13 of 35 in wild type (WT), p < 0.01) and Mac-1(-/-) (17 of 34 vs 6 of 25, p < 0.01) mice . All deaths in LFA-1(-/-) mice occurred after 72 h, whereas most deaths in Mac-1(-/-) mice occurred within 24-48 h . At 24 h, 21 of 27 Mac-1(-/-) mice were bacteremic, vs 15 of 25 WT (p = 0.05); no difference was observed between LFA-1(-/-) and WT . Increased bacteria were recovered from Mac-1(-/-) spleens at 2 h (p = 0.03) and 6 h (p = 0.002) and from livers (p = 0.001) by 6 h . No difference was observed at 2 h in LFA-1(-/-) mice, but by 6 h increased bacteria were recovered from spleens (p = 0.008) and livers (p = 0.04) . Baseline and peak leukocyte counts were similar between Mac-1(-/-) and WT, but elevated in LFA-1(-/-) . At 8 h, peritoneal neutrophils were increased in Mac-1(-/-), but not significantly different in LFA-1(-/-) . Histopathologically, at 24 h Mac-1(-/-) animals had bacteremia and lymphoid depletion, consistent with sepsis . LFA-1(-/-) mice had increased incidence of otitis media and meningitis/encephalitis vs WT at 72 and 96 h . Both Mac-1 and LFA-1 play important but distinct roles in host defense to S . pneumoniae. Vet Microbiol, 2001 Aug 20, 81(4), 331 - 44 Passive immunization of pigs against experimental infection with Streptococcus suis serotype 2; Andresen LO et al.; The safety and protective efficacy of a horse antiserum raised against inactivated whole cell preparations of Streptococcus suis serotype 2 was investigated in pigs by experimental challenge . The antiserum was evaluated in two similar experiments each comprising 12 4-week-old pigs treated with 6 ml of antiserum the day before challenge and four pigs used as challenge controls . Pigs were infected by subcutaneous injection with approximately 10(11) colony forming units of S . suis serotype 2 . Clinical disease in the pigs that could be attributed to infection with S . suis was reduced from 88 to 35% (P = 0.015) . The percentage of pigs with lesions that could be associated with S . suis was reduced from 88 to 22% (P = 0.002) and isolation of S . suis serotype 2 was reduced from five (63%) out of eight pigs in the combined challenge control groups to 3 (13%) out of 23 pigs in the combined treatment groups . These results indicate that passive immunization of pigs may be a way to reduce or control S . suis serotype 2 infections in pigs. Clin Infect Dis, 2001 Jul 1, 33(1), 16 - 21 Epub 2001 May 23. Pneumococcal infections in children after transplantation; Schutze GE et al.; Bacterial infections in recipients of bone marrow and solid-organ transplants remain a major cause of morbidity and death . The cases of 42 children who had undergone transplantation and developed an infection with Streptococcus pneumoniae were retrospectively reviewed . Thirty-four patients had 1 episode of infection, whereas 7 had 2 episodes and 1 had 3 episodes of infection . Solid-organ recipients were more likely to have recurrent invasive disease (P<.02) . A total of 31 (74%) of 42 patients were on immunosuppressive therapy, and 74% had been on antimicrobial therapy within 30 days before diagnosis of S . pneumoniae infection . Only 33% of eligible patients had received a pneumococcal vaccine . Twenty-six percent of isolates recovered were not susceptible to penicillin, and 18% were not susceptible to ceftriaxone . Two patients experienced infection-related deaths; one of these had a penicillin-nonsusceptible isolate . The antimicrobial susceptibilities and outcome of infections with S . pneumoniae in patients who have undergone transplantation are similar to those in the general pediatric population. J Antimicrob Chemother, 2001 Jun, 47(6), 875 - 7 Antimicrobial activity of moxifloxacin, gatifloxacin and six fluoroquinolones against Streptococcus pneumoniae; Saravolatz L et al.; The in vitro and pharmacodynamic effects of moxifloxacin and gatifloxacin against Streptococcus pneumoniae were compared with six other fluoroquinolones . Organisms included penicillin-susceptible (54) and penicillin-non-susceptible (145) isolates from 1998-1999 . Moxifloxacin and clinafloxacin demonstrated the greatest in vitro activity, with MIC90s of 0.13 mg/L, followed by trovafloxacin, grepafloxacin > gatifloxacin, sparfloxacin > levofloxacin > ciprofloxacin . There was no difference in fluoroquinolone activity between penicillin-susceptible and -non-susceptible strains . Pharmacodynamic analysis using published pharmacokinetic information indicates that all the agents tested except ciprofloxacin had an AUC/MIC90 > 30, with moxifloxacin having the greatest free-drug availability. J Antimicrob Chemother, 2001 Jun, 47(6), 863 - 6 Distribution of resistance genes tet(M), aph3'-III, catpC194 and the integrase gene of Tn1545 in clinical Streptococcus pneumoniae harbouring erm(B) and mef(A) genes in Spain; Seral C et al.; The most prevalent macrolide resistance phenotype and genotype among pneumococcal isolates was the cMLSB phenotype {erm(B) or erm(B)/mef(A)} (91.3%) . We studied the distribution of other resistance genes, tet(M), catpC194, aph3'-III, in these strains, seeing evolution at work in that some strains carried different combinations of resistance determinants . The most prevalent patterns associated with resistance to erythromycin {erm(B)} were resistance to tetracycline {tet(M)} and chloramphenicol (catpC194) (48.2%) or resistance to tetracycline {tet(M)} alone (42.2%) . In our isolates of Streptococcus pneumoniae there was a strong association of the erm(B) and tet(M) genes with Tn1545-related elements. J Antimicrob Chemother, 2001 Jun, 47(6), 837 - 40 Lung concentrations of telithromycin after oral dosing; Khair OA et al.; Concentrations of telithromycin were measured in plasma, bronchial mucosa (BM), epithelial lining fluid (ELF) and alveolar macrophages (AM) following multiple oral doses . Concentrations were determined using a microbiological assay . There were 20 subjects in the study, allocated to three nominal time periods: 2, 12 and 24 h . Mean concentrations in plasma, BM, ELF and AM for 2, 12 and 24 h were as follows: 2 h, 1.86 mg/L, 3.88 mg/kg, 14.89 mg/L and 69.32 mg/L; 12 h, 0.23 mg/L, 1.41 mg/kg, 3.27 mg/L and 318.1 mg/L; and 24 h, 0.08 mg/L, 0.78 mg/kg, 0.97 mg/L and 161.57 mg/L . These concentrations of telithromycin in BM and ELF exceeded for 24 h the mean MIC90s of the common respiratory pathogens Streptococcus pneumoniae (0.12 mg/L) and Moraxella catarrhalis (0.03 mg/L), as well as the atypical microorganism Mycoplasma pneumoniae (0.001 mg/L), and suggest that telithromycin may be effective for the treatment of community-acquired pneumonia and chronic obstructive pulmonary disease. J Antimicrob Chemother, 2001 Jun, 47(6), 811 - 8 Pharmacodynamics of moxifloxacin, levofloxacin and sparfloxacin against Streptococcus pneumoniae; Lister PD et al.; An in vitro pharmacokinetic model (IVPM) was used to simulate the human serum pharmacokinetics of moxifloxacin, levofloxacin and sparfloxacin, and to compare their pharmacodynamics against Streptococcus pneumoniae exhibiting a wide range of susceptibilities to fluoroquinolones . Logarithmic-phase cultures were exposed to peak concentrations achieved in human serum of moxifloxacin, levofloxacin or sparfloxacin with oral doses of 400, 500 and 200 mg, respectively . Human elimination pharmacokinetics were simulated, and viable bacterial counts were measured at 0, 1, 2, 4, 6, 8, 24 and 36 h . Moxifloxacin was rapidly bactericidal (>3 logs of killing) against all 10 S . pneumoniae strains, with 99.9% kills of eight strains occurring within 1-3 h after dosing . Maximum kills ranged from 5 to >6 logs . Moxifloxacin eradicated seven strains from the IVPM within 8 h of the first dose, and eradicated two other strains within 24 h . Although levofloxacin and sparfloxacin were also bactericidal against all 10 S . pneumoniae strains, the rates of killing were somewhat slower, with sparfloxacin exhibiting the slowest rate of kill . In summary, moxifloxacin's increased anti-pneumococcal potency compared with levofloxacin and its more favourable pharmacokinetics compared with sparfloxacin provided enhanced pharmacodynamic activity against some S . pneumoniae strains when maximum doses were simulated in an IVPM. J Biochem (Tokyo), 2001 Jun, 129(6), 923 - 8 Molecular cloning and expression of endo-beta-N-acetylglucosaminidase D, which acts on the core structure of complex type asparagine-linked oligosaccharides; Muramatsu H et al.; Endo-beta-N-acetylglucosaminidase D (Endo D) produced by Streptococcus pneumoniae cleaves the di-N-acetylchitobiose structure in asparagine-linked oligosaccharides . The enzyme generally acts on complex type oligosaccharides after removal of external sugars by neuraminidase, beta-galactosidase, and beta-N-acetylglucosaminidase . We cloned the gene encoding the enzyme and expressed it as a periplasm enzyme in Escherichia coli . The first 37 amino acids in the predicted sequence are removed in the mature enzyme, yielding a protein with a molecular mass of 178 kDa . The substrate specificity of the recombinant enzyme is indistinguishable from the enzyme produced by S . pneumoniae . Endo-beta-N-acetylglucosaminidase A (Endo A) from Arthrobacter protophormiae, the molecular mass of which is 72 kDa, had 32% sequence identity to Endo D, starting from the N-terminal sides of both enzymes, although Endo A hydrolyzes high-mannose-type oligosaccharides and does not hydrolyze complex type ones . Endo D is not related to endo-beta-N-acetylglucosaminidases H, F(1), F(2), or F(3), which share common structural motifs . Therefore, there are two distinct groups of endo-beta-N-acetylglucosaminidases acting on asparagine-linked oligosaccharides . The C-terminal region of Endo D shows homology to beta-galactosidase and beta-N-acetylglucosaminidase from S . pneumoniae and has an LPXTG motif typical of surface-associated proteins of Gram-positive bacteria . It is possible that Endo D is located on the surface of the bacterium and, together with other glycosidases, is involved in virulence. Am Fam Physician, 2001 May 15, 63(10), 1991 - 8 Pneumococcal conjugate vaccine for young children; Zimmerman RK; Streptococcus pneumoniae causes approximately 3,300 cases of meningitis, 100,000 to 135,000 cases of pneumonia requiring hospitalization and 6 million cases of otitis media annually in the United States . Pneumococcal conjugate vaccine, approved in 2000 for use in the United States, was designed to cover the seven serotypes that account for about 80 percent of invasive infections in children younger than six years . This vaccine demonstrated 100 percent efficacy against invasive pneumococcal disease in the primary analysis of a large randomized, double-blind, controlled trial . In the follow-up analysis, performed eight months after the trial ended, efficacy against invasive disease was found to be 94 percent for the included serotypes . When initiated during infancy, the four-dose vaccination schedule is set at two, four, six and 12 to 15 months of age . The American Academy of Family Physicians recommends routine vaccination of infants, catch-up vaccination of children younger than 24 months and catch-up vaccination of children 24 to 59 months of age with high-risk medical conditions such as sickle cell disease and congenital heart disease. Ann Emerg Med, 2001 Jun, 37(6), 711 - 9 Principles of appropriate antibiotic use for acute pharyngitis in adults: background; Cooper RJ et al.; The following principles of appropriate antibiotic use for adults with acute pharyngitis apply to immunocompetent adults without complicated comorbid conditions, such as chronic lung or heart disease, and history of rheumatic fever . They do not apply during known outbreaks of group A streptococcus . 1 . Group A beta-hemolytic streptococcus (GABHS) is the causal agent in approximately 10% of adult cases of pharyngitis . The large majority of adults with acute pharyngitis have a self-limited illness, for which supportive care only is needed . 2 . Antibiotic treatment of adult pharyngitis benefits only those patients with GABHS infection . All patients with pharyngitis should be offered appropriate doses of analgesics and antipyretics, as well as other supportive care . 3 . Limit antibiotic prescriptions to patients who are most likely to have GABHS infection . Clinically screen all adult patients with pharyngitis for the presence of the four Centor criteria: history of fever, tonsillar exudates, no cough, and tender anterior cervical lymphadenopathy (lymphadenitis) . Do not test or treat patients with none or only one of these criteria, since these patients are unlikely to have GABHS infection . For patients with two or more criteria the following strategies are appropriate: (a) Test patients with two, three, or four criteria by using a rapid antigen test, and limit antibiotic therapy to patients with positive test results; (b) test patients with two or three criteria by using a rapid antigen test, and limit antibiotic therapy to patients with positive test results or patients with four criteria; or (c) do not use any diagnostic tests, and limit antibiotic therapy to patients with three or four criteria . 4 . Throat cultures are not recommended for the routine primary evaluation of adults with pharyngitis or for confirmation of negative results on rapid antigen tests when the test sensitivity exceeds 80% . Throat cultures may be indicated as part of investigations of outbreaks of GABHS disease, for monitoring the development and spread of antibiotic resistance, or when such pathogens as gonococcus are being considered . 5 . The preferred antibiotic for treatment of acute GABHS pharyngitis is penicillin, or erythromycin in a penicillin-allergic patient. Ann Emerg Med, 2001 Jun, 37(6), 690 - 7 Principles of appropriate antibiotic use for treatment of acute respiratory tract infections in adults: background, specific aims, and methods; Gonzales R et al.; The need to decrease excess antibiotic use in ambulatory practice has been fueled by the epidemic increase in antibiotic-resistant Streptococcus pneumoniae . The majority of antibiotics prescribed to adults in ambulatory practice in the United States are for acute sinusitis, acute pharyngitis, acute bronchitis, and nonspecific upper respiratory tract infections (including the common cold) . For each of these conditions--especially colds, nonspecific upper respiratory tract infections, and acute bronchitis (for which routine antibiotic treatment is not recommended)--a large proportion of the antibiotics prescribed are unlikely to provide clinical benefit to patients . Because decreasing community use of antibiotics is an important strategy for combating the increase in community-acquired antibiotic-resistant infections, the Centers for Disease Control and Prevention convened a panel of physicians representing the disciplines of internal medicine, family medicine, emergency medicine, and infectious diseases to develop a series of "Principles of Appropriate Antibiotic Use for Treatment of Acute Respiratory Tract Infections in Adults." These principles provide evidence-based recommendations for evaluation and treatment of adults with acute respiratory illnesses.This paper describes the background and specific aims of and methods used to develop these principles . The goal of the principles is to provide clinicians with practical strategies for limiting antibiotic use to the patients who are most likely to benefit from it . These principles should be used in conjunction with effective patient educational campaigns and enhancements to the health care delivery system that facilitate nonantibiotic treatment of the conditions in question. Curr Infect Dis Rep, 2001 Jun, 3(3), 224 - 232 The Role of Quinolones in Upper Respiratory Tract Infections; Grossman RF; Fluoroquinolones are widely used in clinical practice because of their advanced pharmacokinetic properties, potential activity against most bacterial species, excellent clinical responses, and few side effects . Quinolones have no role in the treatment of pharyngitis or simple otitis media . Until recently, the available fluoroquinolones were not indicated for the treatment of acute purulent sinusitis because of their perceived inactivity against Streptococcus pneumoniae . Although not generally considered to be drugs of first choice, older quinolones have efficacy similar to that of cephalosporins and b-lactams in randomized clinical trials . Well-conducted clinical trials have shown that the new fluoroquinolones are as effective as standard comparators in patients with suspected or proven acute bacterial sinusitis and may allow shorter treatment . Ciprofloxacin remains the fluoroquinolone of choice for chronic otitis media and malignant otitis media . The new "respiratory" fluoroquinolones have microbiologic and pharmacokinetic advantages over the older agents . Clinical trials have confirmed clinical activity, but superiority compared with older agents has not been conclusively shown . Trials devised to demonstrate clinical or pharmacoeconomic benefits are still required. Curr Infect Dis Rep, 2001 Jun, 3(3), 217 - 223 Etiology and Management of Acute and Recurrent Group A Streptococcal Tonsillitis; Barzilai A et al.; Tonsillitis is one of the most prevalent infections in children and adolescents . The etiologic agents might be viral or bacterial . About 30% of cases are reported to be of bacterial origin, mainly due to group A Streptococcus (GAS) . Although in most instances GAS tonsillitis is a self-limited disease, antibiotic treatment is recommended, mainly to prevent the suppurative and nonsuppurative poststreptococcal sequelae of acute rheumatic fever and to prevent glomerulonephritis . In this paper we review the current knowledge of the etiology of acute and recurrent GAS tonsillitis, with special emphasis on a recent hypothesis regarding the etiology of bacterial eradication failure . While penicillin V remains the drug of choice for acute tonsillitis, other antibiotics are being approved and recommended for particular indications in both Europe and the United States. J Dairy Sci, 2001 May, 84(5), 1140 - 8 Multiplex polymerase chain reaction assay for simultaneous detection of Staphylococcus aureus and streptococcal causes of bovine mastitis; Phuektes P et al.; To improve diagnosis of mastitis in dairy cattle, a multiplex polymerase chain reaction (PCR) assay was developed for the simultaneous detection of the four major bacterial causes of bovine mastitis, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis . The target sequence was the 16S to 23S rRNA spacer regions . The performance of the assay was examined with 117 milk samples collected from a subclinically infected herd, and the diagnostic specificities and sensitivities of the multiplex PCR were compared with conventional culture . PCR was significantly more sensitive than culture for detection of S . aureus and S . uberis, but there were no significant differences in sensitivities between PCR and culture for the detection of S . agalactiae and S . dysgalactiae . The results suggest that this multiplex PCR assay could be used as an alternative method in routine diagnosis for rapid, sensitive, and specific simultaneous detection of S . aureus, S . agalactiae, S . dysgalactiae, and S . uberis in milk samples. Nurse Pract, 2001 May, 26(5), 52, 55 - 6, 59-62 A new conjugate vaccine against pneumococcal disease; Wilson-Adkins M et al.; Streptococcus pneumoniae (pneumococcus) is a major cause of morbidity and mortality, particularly among infants and children . Pneumococcal 7-valent conjugate vaccine (PNCRM7) is the first conjugate vaccine known to prevent most invasive pneumococcal disease in infants and children . PNCRM7, which has a favorable safety profile, provides protection against invasive disease caused by antibiotic-resistant strains of S . pneumoniae, and the vaccine has demonstrated a significant impact on otitis media recurrence . Routine immunization with this vaccine should substantially reduce morbidity and mortality and improve the quality of life for infants, children, and their families. Microbiol Mol Biol Rev, 2001 Jun, 65(2), 187 - 207 ; first page, table of contents Pneumococcal virulence factors: structure and function; Jedrzejas MJ; The overall goal for this review is to summarize the current body of knowledge about the structure and function of major known antigens of Streptococcus pneumoniae, a major gram-positive bacterial pathogen of humans . This information is then related to the role of these proteins in pneumococcal pathogenesis and in the development of new vaccines and/or other antimicrobial agents . S . pneumoniae is the most common cause of fatal community-acquired pneumonia in the elderly and is also one of the most common causes of middle ear infections and meningitis in children . The present vaccine for the pneumococcus consists of a mixture of 23 different capsular polysaccharides . While this vaccine is very effective in young adults, who are normally at low risk of serious disease, it is only about 60% effective in the elderly . In children younger than 2 years the vaccine is ineffective and is not recommended due to the inability of this age group to mount an antibody response to the pneumococcal polysaccharides . Antimicrobial drugs such as penicillin have diminished the risk from pneumococcal disease . Several pneumococcal proteins including pneumococcal surface proteins A and C, hyaluronate lyase, pneumolysin, autolysin, pneumococcal surface antigen A, choline binding protein A, and two neuraminidase enzymes are being investigated as potential vaccine or drug targets . Essentially all of these antigens have been or are being investigated on a structural level in addition to being characterized biochemically . Recently, three-dimensional structures for hyaluronate lyase and pneumococcal surface antigen A became available from X-ray crystallography determinations . Also, modeling studies based on biophysical measurements provided more information about the structures of pneumolysin and pneumococcal surface protein A . Structural and biochemical studies of these pneumococcal virulence factors have facilitated the development of novel antibiotics or protein antigen-based vaccines as an alternative to polysaccharide-based vaccines for the treatment of pneumococcal disease. J Heart Valve Dis, 2001 May, 10(3), 367 - 70 Surgical treatment of prosthetic valve endocarditis with left ventricular-aortic discontinuity: reconstruction of the left ventricular outflow tract with a xenopericardial conduit; Aoyagi S et al.; BACKGROUND AND AIM OF THE STUDY: Aortic prosthetic valve endocarditis (PVE) with annular destruction presents a challenge that requires techniques to eradicate the infection and correct the hemodynamic abnormality . METHODS: Between July 1, 1996 and March 31, 2000, six patients with native or PVE of the aortic valve and aortic annular destruction underwent surgical treatment . Of these patients, three (two men, one woman; mean age 71.0 years) had circumferential annular destruction of the aortic annulus, and formed the basis of this study . The microorganisms responsible for the infection were Streptococcus spp . in two patients and Staphylococcus aureus in one patient . In addition to aggressive debridement of the infected tissue, repair was achieved by reconstruction of the left ventricular outflow tract with a xenopericardial conduit and fixation of the new prosthetic valve to the conduit . RESULTS: One patient with ventricular septal perforation, multiple systemic embolism and sepsis died of low cardiac output syndrome soon after surgery . Two operative survivors were followed up for 9 and 51 months, with no late deaths . No patient has experienced recurrent infection, pericardial patch aneurysm, or prosthetic valve detachment . CONCLUSION: These operative procedures provide easy and secure fixation of the pericardial patch to the healthy tissue under excellent operative view, as well as a sturdy structure for the fixation of the new prosthesis, and complete exclusion of the abscess cavity from the blood stream. Ann Acad Med Singapore, 2001 Mar, 30(2), 199 - 202 Bugs for the next century: the issue of antibiotic resistance; Ang BS; OBJECTIVE: To address the issue of emerging antibiotic resistance and examine which organisms will continue to pose problems in the new century . METHODS: Review of articles pertaining to bacteria recognised for increasing resistance . RESULTS: Changing resistance patterns are correlated with patterns of antibiotic use . This results in fewer effective drugs against "old" established bacteria e.g . gram-positives such as Streptococcus pneumoniae and Staphylococcus aureus . Resistance in gram-negative bacteria is also steadily increasing . Nosocomial gram-negative bacteria are capable of many different resistance mechanisms, often rendering them multiply-resistant . Antibiotic resistance results in morbidity and mortality from treatment failures and increased health care costs . CONCLUSION: Despite extensive research and enormous resources spent, the pace of drug development has not kept up with the development of resistance . As resistance spreads, involving more and more organisms, there is concern that we may be nearing the end of the antimicrobial era . Measures that can and should be taken to counter this threat of antimicrobial resistance include co-ordinated surveillance, rational antibiotic usage, better compliance with infection control and greater use of vaccines. Crit Care Med, 2001 May, 29(5), 1071 - 3 Early diagnosis of retroperitoneal necrotizing fasciitis; Pryor JP et al.; OBJECTIVE: To report survival of retroperitoneal necrotizing fasciitis in an inmunocompromised patient and to demonstrate early clinical signs that may help in the prompt diagnosis and treatment of this severe infection . DESIGN: Case report and literature review . SETTING: An adult, 18-bed intensive care unit within a university hospital . PATIENT: A 38-yr-old man who had undergone an uncomplicated closed hemorrhoidectomy was readmitted to the hospital on postoperative day 5 for erythema around the hemorrhoidectomy and a dirty brown discharge from the wound . INTERVENTIONS: Early diagnosis of retroperitoneal necrotizing fasciitis, wide and repeated debridement, broad-spectrum antibiotics, and eventual abdominal wall reconstruction . MEASUREMENTS AND MAIN RESULTS: This patient manifested periumbilical and bilateral flank erythema, reminiscent of the pattern of ecchymosis seen in cases of retroperitoneal hemorrhage . The findings demonstrate a variation of Cullen's and Grey Turner's sign, most often found in patients with hemorrhagic pancreatitis . An abdominal radiograph revealed a ground glass appearance with radiolucency outlining the bladder, consistent with retroperitoneal air . The chest radiograph showed mediastinal air extending into the neck . Sharp debridement of the retroperitoneal fat, the right anterior rectus sheath, and the right anterior thigh fascia was required to gain control of the infection . Operative cultures grew a mixed flora with Eschericha coli, beta-hemolytic streptococcus, and Bacteroides fragilis predominating . The hospital course was complicated by hemodynamic instability, renal failure, pneumonia, and a pelvic abscess . The patient ultimately survived and underwent abdominal wall reconstruction with mesh . CONCLUSION: Retroperitoneal necrotizing fasciitis is an uncommon soft tissue infection that is often fatal . Early diagnosis in this case was facilitated by the unique clinical findings of a modified Cullen's and Grey Turner's sign . A review of the limited available literature suggests that survival of retroperitoneal fasciitis is possible with prompt debridement and antibiotic therapy. Trends Immunol, 2001 Jun, 22(6), 308 - 11 Distinct types of T-cell help for the induction of a humoral immune response to Streptococcus pneumoniae; Snapper CM et al.; Studies have indicated that purified soluble polysaccharide antigens can elicit T cell-independent Ig responses in vivo, although these responses can be modulated by T cells in a noncognate manner . Relatively little is known, however, concerning the parameters that regulate polysaccharide-specific, as well as protein-specific, Ig isotype responses to an intact extracellular bacterium . Using the murine in vivo humoral response to intact Streptococcus pneumoniae as a model it can be shown that CD4+ T-cell receptor alphabeta+ T cells deliver help for both polysaccharide- and protein-specific Ig responses . However, these responses differ fundamentally in their mechanism of action. Carbohydr Res, 2001 Jun 4, 332(3), 249 - 55 Construction of multivalent sialyl Le(x) from the type Ia group B Streptococcus capsular polysaccharide; Zou W et al.; The type Ia group B Streptococcus (GBSIa) capsular polysaccharide was specifically degraded by partial Smith oxidation of 2,3-diol of the Glc in the backbone to fragments representing asialo core repeating units . Sialylation of these oligomers furnished GBSIa multiple repeating units . One, two and three repeating units of GBSIa were obtained pure, and the higher oligomers were obtained as mixtures . After enzymatic fucosylation oligosaccharides carrying bivalent, trivalent and other multivalent sialyl Le(x) epitopes presented as appendages on an oligolactoside scaffold were obtained. Rev Esp Quimioter, 2001 Mar, 14(1), 55 - 62 {Association between MLS antibiotic and tetracycline resistance genes in Streptococcus pneumoniae}; Seral C et al.; In Streptococcus pneumoniae, resistance to macrolide, lincosamide and streptogramin type B (MLS(B)) antibiotics is mediated by erm(B) and mef(A) determinants . Tetracycline resistance is always associated with resistance to minocycline and is due to the presence of the tet(M) gene . The erm(B) determinant is predominant . We demonstrated that the erm(B) gene could be present with mef(A), which is of streptococcal origin, and msr(A), which is of staphylococcal origin, this being an example of genetic promiscuity . The tet(M) determinant was associated with pneumococci harboring the erm(B) gene, while it was not associated with the strains harboring the mef(A) gene . This association is due to the fact that, in most of the cases, erm(B) and tet(M) reside in the same chromosomal conjugative transposon. J Clin Microbiol, 2001 Jun, 39(6), 2358 - 9 Streptococcal meningitis resulting from contact with an infected horse; Downar J et al.; We report a case of group C streptococcal meningitis in a woman with a history of close animal contact as well as head trauma as a result of a kick by a horse . Blood and cerebrospinal fluid cultures grew Streptococcus equi subsp . zooepidemicus, as did a throat culture taken from the colt that had kicked her 2 weeks prior to admission. J Biol Chem, 2001 Aug 10, 276(32), 30024 - 30 Epub 2001 May 24. Identification, substrate specificity, and inhibition of the Streptococcus pneumoniae beta-ketoacyl-acyl carrier protein synthase III (FabH); Khandekar SS et al.; In the bacterial type II fatty acid synthase system, beta-ketoacyl-acyl carrier protein (ACP) synthase III (FabH) catalyzes the condensation of acetyl-CoA with malonyl-ACP . We have identified, expressed, and characterized the Streptococcus pneumoniae homologue of Escherichia coli FabH . S . pneumoniae FabH is approximately 41, 39, and 38% identical in amino acid sequence to Bacillus subtilis, E . coli, and Hemophilus influenzae FabH, respectively . The His-Asn-Cys catalytic triad present in other FabH molecules is conserved in S . pneumoniae FabH . The apparent K(m) values for acetyl-CoA and malonyl-ACP were determined to be 40.3 and 18.6 microm, respectively . Purified S . pneumoniae FabH preferentially utilized straight short-chain CoA primers . Similar to E . coli FabH, S . pneumoniae FabH was weakly inhibited by thiolactomycin . In contrast, inhibition of S . pneumoniae FabH by the newly developed compound SB418011 was very potent, with an IC(50) value of 0.016 microm . SB418011 also inhibited E . coli and H . influenzae FabH with IC(50) values of 1.2 and 0.59 microm, respectively . The availability of purified and characterized S . pneumoniae FabH will greatly aid in structural studies of this class of essential bacterial enzymes and facilitate the identification of small molecule inhibitors of type II fatty acid synthase with the potential to be novel and potent antibacterial agents active against pathogenic bacteria. Biomaterials, 2001 Jun, 22(12), 1683 - 7 Antibacterial activity of particulate bioglass against supra- and subgingival bacteria; Allan I et al.; Particulate Bioglass is a bioactive material used in the repair of periodontal defects . This material undergoes a series of surface reactions in an aqueous environment which lead to osseointegration . The aim of this study was to determine whether these reactions exerted an antibacterial effect on a range of oral bacteria . Streptococcus sanguis, Streptococcus mutans and Actinomyces viscosus were suspended in nutrient broth (NB), artificial saliva (AS) or Dulbecco's modified eagle medium plus 10% foetal calf serum (DMEM + 10%FCS), with or without particulate Bioglass . All bacteria showed reduced viability following exposure to Bioglass in all the media after 1 h . This antibacterial effect increased after 3 h . Porphyromonas gingivalis, Fusobacterium nucleatum, Prevotella intermedia and Actinobacillus actinomycetemcomitans were suspended in either BM broth or 40% horse serum (HS) in RPMI . A considerable reduction in viability was observed with all bacteria tested, in both media, compared to inert glass controls . In further experiments it was found that the viability of S . sanguis was significantly reduced following exposure to NB pre-incubated with Bioglass . Additionally, it was found that neutralisation of this highly alkaline solution eliminated the antibacterial effect . Moreover, a solution of NB and NaOH (of equivalent pH) exerted an antibacterial effect of similar magnitude to that of the solution pre-incubated with Bioglass . Thus, particulate Bioglass exerts an antibacterial effect on certain oral bacteria, possibly by virtue of the alkaline nature of its surface reactions . This may reduce bacterial colonisation of its surface in vivo. Schweiz Monatsschr Zahnmed, 2000, 110(11), 125 - 30 {Antimicrobial effects of tea tree oil (Melaleuca alternifolia) on oral microorganisms}; Kulik E et al.; The essential oil of Melaleuca alternifolia (tea tree oil) exhibits antimicrobial activity against a wide range of Gram-positive and Gram-negative bacteria, yeasts and fungi . In this study the bacteriostatic and bacteriocidal/fungicidal activity of a tea tree oil solution, of a new tea tree oil (Tebodont) and the respective placebo-gel, of a chlorhexidindigluconate-solution and of PlakOut was tested in vitro against ten different oral microorganisms . Minimum inhibitory concentrations were in the range from 0.0293% to 1.25% for the tea tree oil solution and from 0.0082% to 1.25% for the tea tree oil gel . The values for minimum bacteriocidal/fungicidal concentrations were in the range from 0.0521% to 2.5% for the tea tree oil solution and from <0.0098% to 3.33% for the tea tree oil gel . The most susceptible microorganisms were Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis, whereas Streptococcus mutans and Prevotella intermedia were the least susceptible ones . Both for the chlorhexidindigluconate solution and for PlakOut the values for the minimal inhibitory concentration and for the minimal cidal concentration were between <0.0002% and 0.0125%. Rev Pneumol Clin, 2001 Feb, 57(1 Pt 1), 38 - 40 {Lemierre's syndrome: a case report}; Lemiale V et al.; We report a case of Lemierre's syndrome with a pleuropulmonary complication . Lemierre's syndrome is a rare etiology of lung abscess . The diagnosis is clinical and microbiological (anaerobic organisms) . This syndrome associates an acute oropharyngeal infection with septic thrombophlebitis of the internal jugular vein (sometimes many days before the lung lesion) and pulmonary abscess formation . Clinicians should be aware of this syndrome that is fatal in 10% of patients, usually after delayed or missed diagnosis . The frequency of Lemierre's syndrome would be higher if antibiotics were given only to pharyngitis patients positive for streptococcus. Anal Biochem, 2001 Jun 1, 293(1), 88 - 95 Development of an internally quenched fluorescent substrate and a continuous fluorimetric assay for Streptococcus pneumoniae signal peptidase I; Peng SB et al.; Signal peptidase (SPase) I is responsible for the cleavage of signal peptides of many secreted proteins in bacteria and serves as a potential target for the development of novel antibacterial agents due to its unique physiological and biochemical properties . In this paper, we describe a novel fluorogenic substrate, KLTFGTVK(Abz)PVQAIAGY(NO2)EWL, in which 2-aminobenzoic acid (Abz) and 3-nitrotyrosine (Y(NO2)) were used as the fluorescent donor and acceptor, respectively . The substrate can be cleaved by both Streptococcus pneumoniae and Escherichia coli SPase I . Upon cleavage of the fluorogenic substrate by SPase I, the fluorescent intensity increases and can be monitored continuously by spectrofluorometer . Kinetic analysis with S . pneumoniae SPase I demonstrated that the K(m) value for the substrate is 118.1 microM, and the k(cat) value is 0.032 s(-1) . Mass spectrometric analysis and peptide sequencing of the two cleaved products confirmed that the cleavage occurs specifically at the predicted site . More interestingly, the positively charged lysine in the N-terminus of the substrate was demonstrated to be important for effective cleavage . Phospholipids were found to stimulate the cleavage reaction . This stimulation by phospholipids is dependent upon the N-terminal charge of the substrate, indicating that the interaction of the positively charged substrate with anionic phospholipids is important for maintaining the substrate in certain conformation for cleavage . The substrate and assay described here can be readily automated and utilized for the identification of potential antibacterial agents . South Med J, 2001 May, 94(5), 508 - 11 Streptococcus agalactiae endocarditis and giant pyomyoma simulating ovarian cancer; Genta PR et al.; Group B streptococcus (Streptococcus agalactiae) is a common etiology of bacteremia among adults . Pyomyoma is a rare infectious complication of uterine leiomyomas . We report the case of a diabetic postmenopausal woman with a giant pyomyoma simulating an ovarian cancer . It was associated with S . agalactiae endocarditis and deep venous thrombosis of the right external iliac and femoral veins . Treated initially with intravenous penicillin, amikacin, and anticoagulation, the patient later had abdominal hysterectomy with an uneventful recovery . We also review the cases of pyomyoma reported since 1945 . Of 14 cases described (including ours), mortality was 21% . Endocarditis was never reported in association with pyomyoma . The presence of bacteremia and a leiomyoma should raise suspicion for this disease. Zhonghua Yi Xue Za Zhi, 2000 Oct, 80(10), 749 - 52 {Bacterial resistance in streptococcus pneumoniae}; Wang F et al.; OBJECTIVE: To investigate the prevalence of Streptococcus pneumoniae in patients with respiratory tract infection and healthy children and to determine the antibiotic susceptibility in S . pneumoniae . METHODS: Sputum and throat swabs were collected from the adult and children patients . Nasopharyngeal specimens were collected from healthy children with aseptic swab and cultured for S . pneumoniae . Antibiotic susceptibility was determined by agar dilution method . RESULTS: The isolation rates of S . pneumoniae were 10% in both children and adult out-patients, being 30.2% in 2-6 year-old healthy children . Among 82 clinical isolates, only one was low-level resistant to penicillin (MIC 0.125 mg/L) . In 228 S . pneumoniae strains from healthy children, 32 were low-level resistant to penicillin (MIC 0.125-0.5 mg/L), the resistant rate being 14.0% . S . pneumoniae was usually resistant to erythromycin, clindamycin and SMZ-TMP . CONCLUSION: The penicillin-resistant strains are far more resistant to all the agents tested than susceptible strains except fluoroquinolones and vancomycin; in addition, several strains resistant to the third generation cephalosporins are found. Protein Sci, 2001 Jun, 10(6), 1268 - 73 Structural features of a zinc binding site in the superantigen strepococcal pyrogenic exotoxin A (SpeA1): implications for MHC class II recognition; Baker M et al.; Streptococcal pyrogenic exotoxin A (SpeA) is produced by Streptococcus pyogenes, and has been associated with severe infections such as scarlet fever and Streptococcal Toxic Shock Syndrome (STSS) . In this study, the crystal structure of SpeA1 (the product of speA allele 1) in the presence of 2.5 mM zinc was determined at 2.8 A resolution . The protein crystallizes in the orthorhombic space group P2(1)2(1)2, with four molecules in the crystallographic asymmetric unit . The final structure has a crystallographic R-factor of 21.4% for 7,031 protein atoms, 143 water molecules, and 4 zinc atoms (one zinc atom per molecule) . Four protein ligands-Glu 33, Asp 77, His 106, and His 110-form a zinc binding site that is similar to the one observed in a related superantigen, staphylococcoal enterotoxin C2 . Mutant toxin forms substituting Ala for each of the zinc binding residues were generated . The affinity of these mutants for zinc ion confirms the composition of this metal binding site . The implications of zinc binding to SpeA1 for MHC class II recognition are explored using a molecular modeling approach . The results indicate that, despite their common overall architecture, superantigens appear to have multiple ways of complex formation with MHC class II molecules. Arch Biochem Biophys, 2001 Jun 1, 390(1), 128 - 36 Membranotropic effects of the antibacterial agent Triclosan; Villalain J et al.; Triclosan is a broad-spectrum hydrophobic antibacterial agent used in dermatological preparations and oral hygiene products . To gain further insight into the mode of action of Triclosan we examined its effects on membranes by performing leakage titrations of different oral bacteria and studying its interaction with model membranes through the use of different biophysical techniques . There was negligible efflux of intracellular material from Streptococcus sobrinus at the minimal inhibitory concentration of Triclosan; whatever leakage did occur commenced only at much higher concentrations . In contrast, no leakage was observed at even the minimal bactericidal concentration for Porphyromonas gingivalis . Triclosan decreased the onset temperature of the gel to liquid-crystalline phase transition of 1,2-dimyristoyl-sn-glycero-3-phosphocholine and 1,2-dimyristoyl-sn-3-{phospho-rac-glycerol} membranes and was immiscible with these lipids in the fluid phase at concentrations greater than 5 mol% . Steady-state fluorescence anisotropy measurements of different phospholipid/Triclosan samples using 3-(p-6-phenyl-1,3,5-hexatrienyl)-phenylpropionic acid were consistent with the calorimetric data . Incorporation of increasing amounts of Triclosan into 1,2-dielaidoyl-sn-glycero-3-phosphoethanolamine (DEPE) vesicles induced the nonlamellar H(II) hexagonal phase at low temperatures and new immiscible phases at temperatures below the main transition of DEPE . Taking these results together suggests that the antibacterial effects of Triclosan are mediated at least in part through its membranotropic effects, resulting in destabilized structures which compromise the functional integrity of cell membranes without inducing cell lysis . Pediatr Infect Dis J, 2001 May, 20(5), 488 - 94 Linezolid for the treatment of community-acquired pneumonia in hospitalized children . Linezolid Pediatric Pneumonia Study Group; Kaplan SL et al.; OBJECTIVE: To determine the safety, tolerance, pharmacokinetics and efficacy of linezolid, a new oxazolidinone antibiotic in the treatment of community-acquired pneumonia in hospitalized children . DESIGN: A Phase II, open label multicenter study of intravenous linezolid followed by oral linezolid suspension, both at a dose of 10 mg/kg every 12 h . Efficacy was assessed at 7 to 14 days after the last dose of linezolid . PATIENTS: Children 12 months to 17 years old with community-acquired pneumonia admitted to the hospital of 14 participating centers . RESULTS: From July 21, 1998, through May 14, 1999, 79 children were enrolled and 78 received linezolid . Sixty-six children completed treatment and follow-up and were evaluable for clinical outcome . The median age of the evaluable patients was 3 years (range, 1 to 12 years); 47 were 2 to 6 years old . Pathogens were isolated from blood or pleural fluid cultures in 8 children: Streptococcus pneumoniae, 6 (2 penicillin-resistant); Group A Streptococcus, 1; methicillin-resistant Staphylococcus aureus, 1 . Chest tubes were placed in 9 patients . The mean total duration of intravenous and oral administration was 12.2 +/- 6.2 days (range, 6 to 41 days) . The mean peak and trough plasma concentrations of linezolid were 9.5 +/- 4.8 and 0.8 +/- 1.2 microg/ml, respectively . At the follow-up visit 7 to 14 days after the last dose of linezolid, 61 patients (92.4%) were considered cured including all the patients with proven pneumococcal pneumonia, one failed (methicillin-resistant Staphylococcus aureus) and 4 were considered indeterminate . The most common adverse effects in the intent to treat group were diarrhea (10.3%), neutropenia (6.4%) and elevation in alanine aminotransferase (6.4%) . CONCLUSIONS: Linezolid was well-tolerated and could be considered an alternative to vancomycin for treating serious infections caused by antibiotic-resistant Gram-positive cocci in children pending results of additional studies. Pediatr Infect Dis J, 2001 May, 20(5), 482 - 7 Antibody response to the pneumococcal proteins pneumococcal surface adhesin A and pneumolysin in children with acute otitis media; Rapola S et al.; BACKGROUND: Pneumococcal surface adhesin A (PsaA) and pneumolysin (Ply) are common to virtually all Streptococcus pneumoniae isolates, and they are immunogenic and protective against pneumococcal challenge in experimental animals . We have recently shown production of antibodies to PsaA and Ply in young children, but data on the immune response to these antigens during culture-confirmed pneumococcal infection are lacking . OBJECTIVES: To evaluate whether young children respond to S . pneumoniae by producing antibodies to PsaA and Ply during acute otitis media (AOM) . SUBJECTS AND METHODS: A cohort of 329 children was followed prospectively from the age of 2 months to the age of 2 years . Paired sera were obtained during episodes of AOM and used to measure antibodies to PsaA and Ply by enzyme-linked immunosorbent assay . S . pneumoniae cultured from the middle ear fluid was taken as evidence of pneumococcal AOM . The presence of S . pneumoniae in the nasopharyngeal aspirate collected in connection of AOM or any other respiratory infection or in the nasopharyngeal swab collected at scheduled visits was taken to indicate pneumococcal carriage and thus a history of previous contact with S . pneumoniae . RESULTS: Children with previous pneumococcal contacts had high anti-PsaA and anti-Ply concentrations in the acute phase sera regardless of the nature (AOM or carriage) of the current pneumococcal contact . Of the children with no previous pneumococcal contact, those with current pneumococcal AOM had lower antibody concentrations than those with current pneumococcal carriage only . Anti-PsaA and anti-Ply responses were found in children with current pneumococcal contact . The antibody response was strongly associated with low acute phase antibody concentration, but not significantly with age and the nature of the current pneumococcal contact . CONCLUSIONS: We showed that infants are capable of developing a specific antibody response to the pneumococcal proteins PsaA and Ply during AOM. Zhongguo Zhong Xi Yi Jie He Za Zhi, 1998 Feb, 18(2), 92 - 4 {Clinical and experimental studies in treating infantile acute respiratory tract infection with feiyan chuansou oral liquid}; Peng Z et al.; OBJECTIVE: To study the therapeutical mechanism of Feiyan Chuansou Oral Liquid (FCOL) in treating infantile acute respiratory trct infection . METHODS: Clinical and experimental studies with FCOL in treating infantile acute respiratory tract infection . RESULTS: The clinical result showed that the antitussive, expectorant, anti-asthmatic effect and resolution of dry and moist rale and wheezing of FCOL were significantly better in treatment group than those in the control group (P < 0.05) . Animal experimental results also showed that antitussive, expectorant, anti-asthmatic effect of FCOL were significantly better than those of the control group (P < 0.05-0.001) . Bacteriostatic test in vitro showed that FCOL could inhibit streptococcus pneumoniae, klebsiella pneumoniae, influenza bacilli and staphylococcus aureus . Antivirologic test showed that FCOL could inhibit completely the influenza A virus and respiratory syncytial virus . CONCLUSION: FCOL is an effective preparation in treating infantile acute respiratory infection. Int Immunopharmacol, 2001 Mar, 1(3), 433 - 43 Multiple ligand binding sites on domain seven of human complement factor H; Giannakis E et al.; Foreign particles and damaged host cells can activate the complement system leading to their destruction by the host defense system . Factor H (fH) plays a vital role in restricting complement activation on host cells through interactions with polyanions such as heparin, while allowing activation to proceed on foreign surfaces . Complement activation by damaged host cells is also down regulated by fH, which is localized to injured areas through interactions with C-reactive protein (CRP) . A number of pathogens have developed mechanisms by which they can also bind fH and thus exploit its protective properties . One such organism is Group A Streptococcus (GAS) which mediates fH binding via its surface expressed M-protein . fH consists of 20 conserved short consensus repeat (SCR) units and mutagenesis studies indicate that the seventh repeat is responsible for interactions with heparin, CRP and M-protein . We recently performed molecular modelling of fH SCR 7 and identified a cluster of positively charged residues on one face of the domain . By alanine replacement mutagenesis, we demonstrated that these residues are involved in heparin, CRP and M protein binding, which indicates that there is a common site within fH SCR 7 responsible for multiple ligand recognition. AIDS Patient Care STDS, 1996 Dec, 10(6), 336 - 41 Invasive pneumococcal infections in children infected with HIV are not associated with splenic dysfunction; Andiman WA et al.; OBJECTIVES: Children infected with HIV-1 are more likely to acquire infections associated with the encapsulated bacterial pathogens of childhood than their non-HIV-infected peers . The goal of the current study was to determine what proportion of community-acquired, invasive pneumococcal disease in HIV-infected children could be attributed to splenic dysfunction, as measured by enumerating the number of pocked red blood cells (RBCs) in peripheral blood . METHODS: Splenic reticuloendothelial function was assessed semiquantitatively by examining the morphology of the RBCs of 84 children born to HIV-infected mothers using phase interference microscopy . Surveillance of medical records, and a review of the Yale-New Haven Hospital Clinical Microbiology computerized database, revealed that all of the bacterial cultures of blood and cerebrospinal fluid from these patients were positive . RESULTS: Of the 84 children assessed, 70 were infected with HIV (median age 66 months) and 14 were uninfected seroreverters (controls) . Sixty-one of the 70 HIV-infected children met the CDC criteria for moderate or severe immunodeficiency and/or moderate or severe symptomatic conditions . Seventeen of the 70 HIV-infected children experienced 23 invasive bacterial infections . Streptococcus pneumoniae was responsible for all 23 infections . The median age at the time of first infection among these 17 subjects was 20 months (range, 10-58 months) . There were no episodes of invasive bacterial infections in the remaining 53 HIV-infected children nor among the 14 controls . All 84 children studied, including those with invasive pneumococcal disease, had normal proportions (i.e., < 2%) of pocked erythrocytes in peripheral blood . CONCLUSION: Splenic dysfunction, as measured by the pocked RBC count, does not account for the increased occurrence of invasive pneumococcal disease found in children infected with HIV. Arch Biochem Biophys, 2001 Apr 1, 388(1), 165 - 70 Purification and characterization of the single-stranded DNA binding protein from Streptococcus pneumoniae; Steffen SE et al.; The Escherichia coli single-stranded DNA binding (SSB) protein is a non-sequence-specific DNA binding protein that functions as an accessory factor for the RecA protein-promoted three-strand exchange reaction . An open reading frame encoding a protein similar in size and sequence to the E . coli SSB protein has been identified in the Streptococcus pneumoniae genome . The open reading frame has been cloned, an overexpression system has been developed, and the protein has been purified to greater than 99% homogeneity . The purified protein binds to ssDNA in a manner similar to that of the E . coli SSB protein . The protein also stimulates the S . pneumoniae RecA protein and E . coli RecA protein-promoted strand exchange reactions to an extent similar to that observed with the E . coli SSB protein . These results indicate that the protein is the S . pneumoniae analog of the E . coli SSB protein . The availability of highly-purified S . pneumoniae SSB protein will facilitate the study of the molecular mechanisms of RecA protein-mediated transformational recombination in S . pneumoniae. J Immunol, 2001 Jun 1, 166(11), 6711 - 9 Immunological and biochemical characterization of streptococcal pyrogenic exotoxins I and J (SPE-I and SPE-J) from Streptococcus pyogenes; Proft T et al.; Recently, we described the identification of novel streptococcal superantigens (SAgs) by mining the Streptococcus pyogenes M1 genome database at Oklahoma University . Here, we report the cloning, expression, and functional analysis of streptococcal pyrogenic exotoxin (SPE)-J and another novel SAg (SPE-I) . SPE-I is most closely related to SPE-H and staphylococcal enterotoxin I, whereas SPE-J is most closely related to SPE-C . Recombinant forms of SPE-I and SPE-J were mitogenic for PBL, both reaching half maximum responses at 0.1 pg/ml . Evidence from binding studies and cell aggregation assays using a human B-lymphoblastoid cell line (LG-2) suggests that both toxins exclusively bind to the polymorphic MHC class II beta-chain in a zinc-dependent mode but not to the generic MHC class II alpha-chain . The results from analysis by light scattering indicate that SPE-J exists as a dimer in solution above concentrations of 4.0 mg/ml . Moreover, SPE-J induced a rapid homotypic aggregation of LG-2 cells, suggesting that this toxin might cross-link MHC class II molecules on the cell surface by building tetramers of the type HLA-DRbeta-SPE-J-SPE-J-HLA-DRbeta . SPE-I preferably stimulates T cells bearing the Vbeta18.1 TCR, which is not targeted by any other known SAG: SPE-J almost exclusively stimulates Vbeta2.1 T cells, a Vbeta that is targeted by several other streptococcal SAgs, suggesting a specific role for this T cell subpopulation in immune defense . Despite a primary sequence diversity of 51%, SPE-J is functionally indistinguishable from SPE-C and might play a role in streptococcal disease, which has previously been addressed to SPE-C. Mol Microbiol, 2001 May, 40(3), 572 - 85 A Streptococcus pneumoniae pathogenicity island encoding an ABC transporter involved in iron uptake and virulence; Brown JS et al.; Restricted iron availability is a major obstacle to growth and survival of pathogenic bacteria during infection . In contrast to Gram-negative pathogens, little is known about how Gram-positive pathogens obtain this essential metal . We have identified two Streptococcus pneumoniae genetic loci, pit1 and pit2, encoding homologues of ABC iron transporters that are required for iron uptake by this organism . S . pneumoniae strains containing disrupted copies of either pit1 or pit2 had decreased sensitivity to the iron-dependent antibiotic streptonigrin, and a strain containing disrupted copies of both pit1 and pit2 was unable to use haemoglobin as an iron source and had a reduced rate of iron uptake . The pit2- strain was moderately and the pit1-/pit2- strain strongly attenuated in virulence in mouse models of pulmonary and systemic infection, showing that the pit loci play a critical role during in vivo growth of S . pneumoniae . The pit2 locus is contained within a 27 kb region of chromosomal DNA that has several features of Gram-negative bacterial pathogenicity islands . This probable pathogenicity island (PPI-1) is the first to be described for S . pneumoniae, and its acquisition is likely to have played a significant role in the evolution of this important human pathogen. Mol Microbiol, 2001 May, 40(3), 555 - 71 A functional genomic analysis of type 3 Streptococcus pneumoniae virulence; Lau GW et al.; Streptococcus pneumoniae remains a serious cause of morbidity and mortality in humans, but relatively little is known about the molecular basis of its pathogenesis . We used signature-tagged mutagenesis together with an analysis of S . pneumoniae genome sequence to identify and characterize genes required for pathogenesis . A library of signature-tagged mutants was created by insertion-duplication mutagenesis, and 1786 strains were analysed for their inability to survive and replicate in murine models of pneumonia and bacteraemia . One hundred and eighty-six mutant strains were identified as attenuated, and 56 were selected for further genetic characterization based on their ability to excise the integrated plasmid spontaneously . The genomic DNA inserts of the plasmids were cloned in Escherichia coli and sequenced . These sequences were subjected to database searches, including the S . pneumoniae genome sequence, which allowed us to examine the chromosomal regions flanking these genes . Most of the insertions were in probable operons, but no pathogenicity islands were found . Forty-two novel virulence loci were identified . Five strains mutated in genes involved in gene regulation, cation transport or stress tolerance were shown to be highly attenuated when tested individually in a murine respiratory tract infection model . Additional experiments also suggest that induction of competence for genetic transformation has a role in virulence. Glycobiology, 2001 Apr, 11(4), 297 - 304 Kinetic properties of Streptococcus pneumoniae hyaluronate lyase; Kelly SJ et al.; Streptococcus pneumoniae hyaluronate lyase is a surface antigen of this bacterial pathogen, which causes significant mortality and morbidity in human populations worldwide . The primary function of this enzyme is the degradation of hyaluronan, a major component of the extracellular matrix of the tissues of practically all vertebrates . The enzyme uses a processive mode of action to degrade hyaluronan to a final product, an unsaturated disaccharide hyaluronan unit . This catalysis proceeds via a five-step proton acceptance and donation mechanism that includes substrate binding, catalysis, release of the disaccharide product, translocation of the remaining hyaluronan substrate, and proton exchange with microenvironment . Based on the analysis of the three-dimensional structure of the native enzyme and its complexes with hexasaccharide substrate and disaccharide product, several residues have been chosen for mutation studies . These mutated residues included the catalytic residues Asn349, His399, Tyr408, and residues responsible for substrate binding and translocation, Arg243 and Asn580 . The comparison of the kinetic properties of the wild-type with the mutant enzymes allowed for the characterization of every mutant and the correlation of the kinetic properties of the enzyme with its structure . The comparison of the wild-type hyaluronate lyase with other polysaccharide-degrading enzymes, the hydrolases endonuclease and glucoamylase, shows striking similarity of K(m)s for all of these different enzymes. Surg Neurol, 2001 Apr, 55(4), 235 - 9 Frontal sinus osteoma associated with cerebral abscess formation: a case report; Summers LE et al.; BACKGROUND: Osteomas of the paranasal sinuses rarely lead to intracranial manifestations . We present an unusual case of a frontal sinus osteoma leading to intracerebral abscess formation . CASE DESCRIPTION: A 51-year-old Hispanic man presented with increasing frontal headaches, new onset seizure, lethargy, global dysphasia, and unilateral hemiparesis . CSF studies demonstrated mild pleocytosis . Neuroradiological studies revealed an opacity filling the left frontal sinus, as well as a ring-enhancing mass with surrounding edema in the left frontal lobe . The patient was surgically treated with a left frontal osteoplastic craniotomy and removal of the abscess and bony mass . Intraoperative cultures were positive for Streptococcus pneumoniae . Pathology revealed bony tumor consistent with osteoma . The patient's neurological status improved to baseline after surgery . CONCLUSION: The frontal sinus osteoma was associated with rapid development of a frontal lobe abscess, requiring emergent surgical debridement . Although rare, intracerebral manifestations should be considered and expected as a cause of new neurological deficits in the presence of paranasal sinus osteoma. Oral Microbiol Immunol, 2001 Jun, 16(3), 182 - 4 Amine and tin fluoride inhibition of Streptococcus sanguis adhesion under continuous flow; Embleton JV et al.; This study evaluated the ability of topically applied amine fluoride (AmF) and AmF-tin fluoride to inhibit the adhesion of Streptococcus sanguis within a parallel plate flow cell system . One of three AmF compounds and two tin fluoride preparations significantly reduced the net bacterial adhesion to conditioned glass over a 1-h period . Tin(IV) fluoride inhibited S . sanguis adhesion to the greatest extent, and this was shown to be dependent on the formation of the conditioning film prior to agent application. Oral Microbiol Immunol, 2001 Jun, 16(3), 170 - 7 Long-term persistence and recall of immune responses in aged mice after mucosal immunization; Harrod T et al.; To evaluate the retention of memory in the mucosal immune system of aged animals, 2-year-old mice that had been immunized intragastrically at 3 months of age with Streptococcus mutans protein AgI/II coupled to the B subunit of cholera toxin (CTB) were evaluated by ELISA for antibodies to AgI/II and CT in serum, saliva, and vaginal wash . To evaluate recall responses, mice were then immunized intragastrically with AgI/II-CTB, in comparison with previously unimmunized controls . Those that had been primed in their youth showed a more rapid antibody response in serum (immunoglobulin G (IgG)) and secretions (IgA), but all animals eventually responded to a similar degree after the third dose . Mice immunized at 3 months also retained for 2 years spleen cells capable of proliferating in vitro in response to AgI/II . These data show that aged mice retain the ability to mount immune responses to mucosally presented immunogens and that memory to mucosally presented immunogens can persist for almost the whole lifetime of a mouse. Pediatr Nephrol, 2001 Apr, 16(4), 362 - 5 Prognosis of Streptococcus pneumoniae-induced hemolytic uremic syndrome; Nathanson S et al.; Streptococcus pneumoniae-induced hemolytic uremic syndrome (HUS) is known to be a severe acute disease leading to death in one-third of cases, but data regarding the long-term follow-up are lacking . A new series of 11 patients with Streptococcus pneumoniae-induced HUS associated with meningitis and pneumonia constituted a multi-center review . Among 9 patients with a severe acute infectious disease, 3 died from meningitis and 1 from neurological sequelae after a partial recovery of renal function . The mean duration of dialysis was 32 days in patients with acute renal failure who survived the acute infectious period . Cortical necrosis was documented in five of six kidney specimens . Among the 7 surviving patients, 5 developed end-stage renal failure 4-17 years later. Antimicrob Agents Chemother, 2001 Jun, 45(6), 1911 - 4 Evaluation of susceptibility testing to detect fluoroquinolone resistance mechanisms in Streptococcus pneumoniae; Richardson DC et al.; To determine if susceptibility testing of Streptococcus pneumoniae could detect those isolates that had one of the recognized mechanisms of fluoroquinolone resistance, 101 isolates were selected; the levofloxacin MIC for 28 of these isolates was > or =4 microg/ml . Only isolates with both parC and gyrA mutations or with no recognized resistance mechanisms were reliably identified by using these results . Isolates with only a parC mutation could not be detected reliably using any susceptibility testing method. Antimicrob Agents Chemother, 2001 Jun, 45(6), 1788 - 93 Inhibition of group A streptococcus infection by carboxyfullerene; Tsao N et al.; The effect of a water-soluble trimalonic acid derivative of fullerene, carboxyfullerene, against Streptococcus pyogenes infection was tested . Pretreatment with carboxyfullerene was able to protect mice from S . pyogenes infection in an air pouch model . S . pyogenes-induced death and skin injury were inhibited dose dependently by carboxyfullerene . Administration of carboxyfullerene via the peritoneum and air pouch at 3 h post-S . pyogenes infection was able to protect 33% of mice from death . Surveys of exudates of the air pouch of carboxyfullerene-treated mice revealed that survival of infiltrating neutrophils was prolonged and that the bacteria were eliminated as a result of enhanced bactericidal activity of the neutrophils . Furthermore, carboxyfullerene was able to directly inhibit in vitro growth of S . pyogenes . These data suggest that carboxyfullerene can be considered an antimicrobial agent for group A streptococcus infection. Antimicrob Agents Chemother, 2001 Jun, 45(6), 1721 - 9 Antimicrobial resistance among clinical isolates of Streptococcus pneumoniae in the United States during 1999--2000, including a comparison of resistance rates since 1994--1995; Doern GV et al.; A total of 1,531 recent clinical isolates of Streptococcus pneumoniae were collected from 33 medical centers nationwide during the winter of 1999--2000 and characterized at a central laboratory . Of these isolates, 34.2% were penicillin nonsusceptible (MIC > or = 0.12 microg/ml) and 21.5% were high-level resistant (MIC > or = 2 microg/ml) . MICs to all beta-lactam antimicrobials increased as penicillin MICs increased . Resistance rates among non-beta-lactam agents were the following: macrolides, 25.2 to 25.7%; clindamycin, 8.9%; tetracycline, 16.3%; chloramphenicol, 8.3%; and trimethoprim-sulfamethoxazole (TMP-SMX), 30.3% . Resistance to non-beta-lactam agents was higher among penicillin-resistant strains than penicillin-susceptible strains; 22.4% of S . pneumoniae were multiresistant . Resistance to vancomycin and quinupristin-dalfopristin was not detected . Resistance to rifampin was 0.1% . Testing of seven fluoroquinolones resulted in the following rank order of in vitro activity: gemifloxacin > sitafloxacin > moxifloxacin > gatifloxacin > levofloxacin = ciprofloxacin > ofloxacin . For 1.4% of strains, ciprofloxacin MICs were > or = 4 microg/ml . The MIC(90)s (MICs at which 90% of isolates were inhibited) of two ketolides were 0.06 microg/ml (ABT773) and 0.12 microg/ml (telithromycin) . The MIC(90) of linezolid was 2 microg/ml . Overall, antimicrobial resistance was highest among middle ear fluid and sinus isolates of S . pneumoniae; lowest resistance rates were noted with isolates from cerebrospinal fluid and blood . Resistant isolates were most often recovered from children 0 to 5 years of age and from patients in the southeastern United States . This study represents a continuation of two previous national studies, one in 1994--1995 and the other in 1997--1998 . Resistance rates with S . pneumoniae have increased markedly in the United States during the past 5 years . Increases in resistance from 1994--1995 to 1999--2000 for selected antimicrobial agents were as follows: penicillin, 10.6%; erythromycin, 16.1%; tetracycline, 9.0%; TMP-SMX, 9.1%; and chloramphenicol, 4.0%, the increase in multiresistance was 13.3% . Despite awareness and prevention efforts, antimicrobial resistance with S . pneumoniae continues to increase in the United States. Antimicrob Agents Chemother, 2001 Jun, 45(6), 1654 - 9 Activities of newer fluoroquinolones against ciprofloxacin-resistant Streptococcus pneumoniae; Coyle EA et al.; The incidence of ciprofloxacin resistance in Streptococcus pneumoniae is low but steadily increasing, which raises concerns regarding the clinical impact of potential cross-resistance with newer fluoroquinolones . To investigate this problem, we utilized an in vitro pharmacodynamic model and compared the activities of gatifloxacin, grepafloxacin, levofloxacin, moxifloxacin, and trovafloxacin to that of ciprofloxacin again